Autoimmune characteristics of AI-prone MRL-lpr/lpr and control non-autoimmune mice [31].
\r\n\tEqually important are the consequences deriving from the extraordinary nature of the present times. The COVID-19 pandemic and the restrictive measures to contain the infection (lockdown and "physical distancing" in primis) have revolutionized the lives, and a distortion/modification of habits, rhythms, arrangements will continue to be necessary.
\r\n\tGovernments have implemented a series of actions to mitigate the spread of infections and alleviate the consequent pressure on the hospital system. On the other hand, the Covid-19 pandemic has caused a series of other cascading effects that will probably be much more difficult to mitigate and which expose to complex consequences. The past two years have brought many challenges, particularly for healthcare professionals, students, family members of COVID-19 patients, people with mental disorders, the frail, the elderly, and more generally those in disadvantaged socio-economic conditions, and workers whose livelihoods have been threatened. Indeed, the substantial economic impact of the pandemic may hinder progress towards economic growth as well as progress towards social inclusion and mental well-being.
\r\n\t
\r\n\tAlthough in all countries the knowledge on the impact of the pandemic on mental health is still limited and mostly derived from experiences only partially comparable to the current epidemic, such as those referring to the SARS or Ebola epidemics, it is likely that the demand for intervention it will increase significantly in the coming months and years. The extraordinary growth of scientific research in the field of neuroscience now offers the possibility of a new perspective on the relationship between mind and brain and generates new scenarios in understanding the long wave of the pandemic and in the prospects for treatment. Moreover, the pandemic also has led to opportunities to implement remote monitoring and management interventions.
\r\n\t
\r\n\tOverall this volume will address the complex relationship existing between COVID-19, mental health, acquired knowledge, and possible interventions taking a highly multidisciplinary approach; from physiological and psychobiological mechanisms, and neuromodulation through medical treatment, psychosocial interventions, and self-management.
According to classical conception, antibodies (Abs) are specific proteins produced by the immune systems with exclusive function of antigen binding. Antibodies can act similarly to specific enzymes, but in contrast to enzymes, they cannot catalyze chemical conversions of their ligands. For most part of antibodies, this observation is correct. At the same time during the last 30 years, it was shown that Abs against chemically stable analogues modeling the transition states of chemical reaction can catalyze many different reactions [1–8]. Such artificial catalytic antibodies were called abzymes (derived from antibody enzymes). Abzymes (Abzs) can catalyze more than 200 different chemical reactions and are new biological catalysts attracting great interest during recent years and are well described (for review see [1–8] and refs therein).
\nFirst, natural Abzs were found in patients with bronchial asthma; they hydrolyze vasoactive intestinal peptide [9]. Then IgGs hydrolyzing DNA were revealed in the blood of patients with systemic lupus erythematosus (SLE) [10]. The third natural Abzs were SLE IgGs with RNase activity [11]. To date, many catalytic Abs (IgGs and/or IgAs, IgMs) catalyzing hydrolysis of different RNA, DNA, nucleotides, oligopeptides, proteins, lipids, and oligosaccharides were revealed in the sera of patients with various autoimmune and viral diseases (for review, see Refs. [8, 12–22] and refs therein).
\nSome idiotypic Abzs against foreign antigens and auto-Abzs to self-antigens having different catalytic activities may be spontaneously induced by primary antigens simulating in varying degrees the transition states of chemical reactions [8, 12–22]. At the same time, some antiidiotypic Abs against active centers of many enzymes are also catalytically active [8, 12–22, 23–28]. Healthy humans do more often not demonstrate catalytic Abs or their activities are extremely low. It was shown that detection of Abzs is the earliest indicator of different autoimmune diseases (ADs) development [8, 12–22]. At the outset and early stages of ADs analyzed, the repertoire of abzymes is more often relatively narrow, but it is expanding very much with the progress of AI diseases; the generation of diverse Abzs with many various activities and functions may be observed [8, 12–22]. Some abzymes are cytotoxic and dangerous for people; they can play a very important negative role in the different AD pathogenesis [8, 12–22]. However, specific positive roles have been also proposed for several abzymes. Increase in Abzs activities is associated with a specific reorganization of the immune system including change in differentiation profile and level of proliferation of hematopoietic stem cells of bone marrow as well as lymphocyte proliferation in different organs of SLE and experimental autoimmune encephalomyelitis (EAE) in mice [29–32]. Different mechanisms of abzymes production were revealed in healthy animals after external immunization and in autoimmune mice during their spontaneous or antigen-induced development of autoimmune processes [29–32].
\nCatalysis of different reactions by abzymes is potentially important for many different fields; specific reaction for a synthesis of new drugs, which may be useful for therapy, estimation of abzyme’s possible role in innate and adaptive immunity, as well as for understanding of destructive responses and self-tolerance in ADs [33–37].
\nIn this chapter, Abzs with various catalytic activities in SLE are described and compared with abzymes in case of other autoimmune pathologies. In addition, a possible role of different defects of immune systems resulting in changes of differentiation profile of hematopoietic stem cells (HSCs) of mice bone marrow as well as an increase in lymphocyte proliferation in thymus, bone marrow, spleen, and a significant suppression in these organs of cell apoptosis associated with the abzymes production is discussed.
\nAs mentioned above, SLE is systemic polyethiologic diffuse autoimmune disease, symptoms of which for different patients vary significantly and may be from mild to severe. The exact reason and mechanisms of SLE development till yet is not clear [38]. Genetic, environmental, hormonal, and immune factors may play an important role for the development of SLE. Many different autoimmune diseases (ADs) including SLE are characterized by spontaneous generation of primary antibodies to nucleotides, nucleic acids and their complexes, proteins, polypeptides, polysaccharides, etc. [8, 16–22, 36, 38–42]. Anti-DNA auto-Abs without catalytic activities are detectable even in the sera of healthy humans and their relative titres vary from individual to individual significantly [43, 44].
\nSLE is usually considered to be associated with autoimmunization of patients with DNA, since sera of such patients usually contain anti-DNA Abs and DNA in increased concentrations comparing with that for healthy volunteers [38]. However, comparing to healthy donors, anti-DNA antibody concentrations are higher not only in patients with SLE (36% of patients) [43, 45], but also in Hashimoto’s thyroiditis (23%) [43], multiple sclerosis (17–18%) [43, 46, 47], rheumatoid arthritis (7%), myasthenia gravis (6%), and Sjogren’s syndrome (18%) [43]. In addition, from the cloning of the IgG repertoire from directly active plaques and periplaque regions of brain and from B-cells of the cerebrospinal fluid of MS patient, new keys to the understanding of this pathology were proposed [48]. High affinity anti-DNA Abs were shown to be the major components of the intrathecal IgG response. In addition, monoclonal anti-DNA Abs of multiple sclerosis (MS) patients and Abs specific to DNA derived from SLE patients interact efficiently with the surface of neuronal cells and oligodendrocytes [48]. Recognition of cell-surface by these Abs was DNA-dependent. The data indicate that Abs against DNA may be important for autoimmune and neuropathological mechanisms in chronic SLE and MS [48]. Interestingly, SLE and MS patients show some similarity in the same medical, biochemical, and immunological indexes including anti-DNA and other auto-Abs [13–22].
\nAnti-DNA and anti-RNA Abs with DNase and RNase activities were for the first time detected in sera of SLE patients [10, 11] and then with other ADs [13–22]. The origin of natural Abzs is very complex. First, similar to artificial abzymes, they may originate against analogues of transition states of chemical reactions or against enzyme substrates acting as haptens [8–22]. Many antigens can change their conformation after association with various proteins, and in such complexes, their structure could mimic that of a transition state of chemical reaction substrate. For example, DNA is a bad antigen and immunization of animals with pure DNA or RNA leads to the production of abzymes with very low DNase and RNase activities [49, 50]. Many anti-DNA auto-Abs in SLE are directed against DNA-histone nucleosomal complexes, resulting from internucleosomal cleavage during apoptosis [42]. Apoptotic cells and their different components are the primary antigens as well as immunogens in SLE and are important for the recognition, processing, perception, and/or apoptotic autoantigen presentation by antigen-presenting cells during development of autoimmune processes [42]. Therefore, immunization of mice with complex of DNA and histones or positively charged methylated bovine albumin, simulating positively charged histones, results in production of anti-DNA Abs and DNase abzymes with high activity [29–31, 49, 50]. It was shown that abzymes with different activities may be obtained with a significantly higher incidence in autoimmune mouse strains comparing to conventionally used control nonautoimmune mice [51, 52]. Immunization of autoimmune-prone MRL-lpr/lpr mice with DNA-protein complexes also results in significantly higher production of anti-DNA Abs and abzymes with DNase activity comparing with nonautoimmune CBA and BALB/c healthy mice [29–31]. At the same time, artificial antiidiotypic abzymes can be induced by immunization of animals with different enzymes [23–28]. It was first suggested that natural SLE DNase Abzs may be antiidiotypic abzymes to topoisomerase I [53]. Immunization of rabbits with DNase I led to the production of Abzs with DNase activity of antiidiotypic nature [54]. Idiotypic Abs first were obtained by immunization of animals with DNase I and then they were used to elicit a polyclonal antiidiotypic Abs hydrolyzing DNA; it indicates for the existence of internal Abs structure mimicking active centre of DNase I [54]. We have suggested that polyclonal DNase Abzs in autoimmune patients may be a cocktail of abzymes against complexes of proteins with DNA and RNA and antiidiotypic abzymes to different DNA-hydrolyzing enzymes. Therefore, we have immunized rabbits with DNase II, DNase I, pancreatic RNase A, DNA, and RNA [49, 50, 55–57]. In all cases, abzymes with intrinsic DNase and RNase activities were revealed. IgGs against DNase I with DNase activity also have an antiidiotypic nature [55]. Interestingly, 74–85% of the total polyclonal IgGs against RNase A possessing RNase and DNase activities belong to antiidiotypic Abs, while 15–26% of the Abzs cannot interact with affinity sorbent-bearing Abs against RNase A; they bind with DNA- and RNA-Sepharoses and may be antibodies to nucleic acids bound to RNase [56]. In addition, only ~10% of the polyclonal total IgGs demonstrating DNase and RNase activities from sera of rabbits immunized with DNase II have antiidiotypic nature, while the remaining 90% of Abzs did not interact with Sepharose-bearing Abs against DNase II, they may also be Abs to nucleic acids bound to DNase II [57]. The relatively low amount of antiidiotypic abzymes against DNase II hydrolyzing DNA and RNA may be a consequence of low immunogenicity of DNase II active site comparing with other antigenic determinants of this nuclease. Antibodies against DNA and RNA complexes with proteins and other antinuclear components were found in the blood sera of patients with several multisystem connective tissue diseases including SLE [58]. Interestingly, abzymes agains DNA and RNA bound with proteins are usually significantly more active in the hydrolysis of these substrates than antiidiotypic Abzs against enzyme active centres [17–22, 49, 50, 55–57]. Thus, RNase A, DNase I, DNase II, and other DNA- and RNA-dependent enzymes can themselves be antigens producing not only antiidiotypic abzymes with corresponding active sites, but these enzymes can interact with RNA and DNA and induce formation of anti-RNA and or anti-DNA abzymes possessing no affinity for these enzymes, but having higher catalytic activities than antiidiotypic Abzs. In addition, various proteins interacting with DNA and RNA can differ in their ability to produce antiidiotypic Abzs and the formation of abzymes against bound nucleic acids. Overall, it is clear that abzymes of patients with various autoimmune diseases can be very different cocktails of idiotypic antibodies directly against DNA, RNA, and against complexes of these antigens with different enzymes or proteins as well as antiidiotypic Abs against many DNA-dependent enzymes [17–22, 49, 50, 55–57].
\nIt was shown that DNase Abzs of patients with SLE [59], MS [16], and DNA-hydrolyzing Bence-Jones proteins of patients with multiple myeloma [60] are cytotoxic, able to penetrate cell nucleus and cause fragmentation of nuclear DNA leading to cell apoptosis. A significant decrease in cell apoptosis in the case of ADs may be a very important factor providing the increase in the level of specific lymphocytes producing auto-Abs and abzymes, which are usually eliminated in different organs of healthy mammals [61, 62]. The cell apoptosis caused by Abzs with DNase activity leads to increase in the concentration of histones complexes with DNA fragments in the blood of mammals and, consequently, to production of antibodies against DNA and DNA-hydrolyzing abzymes. Thus, the appearance Abzs with DNase activity in the blood of mammals may be a very important factor in the strengthening of the autoimmune reactions [13–22]. The abzymes with DNase activity should be considered as very dangerous since they can stimulate development of autoimmune reactions. The overall level of autoimmune reactions may depend on the ratio of cytotoxic (harmful) and beneficial to organisms auto-Abs. Therefore, it was very interesting to elucidate what factors underlie in the AI processes development and how possible mechanisms of autoimmunity are associated with the production of abzymes. Some data suggest that various ADs can originate from defects in the hematopoietic stem cells (HSCs) [63]. Therefore, it was reasonable to analyze what defects or changes may be revealed in the HSCs during spontaneous and DNA-induced development of SLE in autoimmune prone MRL-lpr/lpr mice.
\nIt is known that after spontaneous development of SLE, MRL-lpr/lpr mice are characterized by visual symptoms of autoimmune pathology (baldness of head and parts of the back, pink spots, general health deterioration, etc.). Appearance of pronounced visual symptoms usually well correlate with high proteinuria (≥3 mg/ml of protein concentration in urine) [64, 65]. It was shown previously that sera of spontaneously diseased MRL-lpr/lpr mice contain Abzs with high DNase activity correlating with high proteinuria and visual symptoms usually at age of 5–12 months [64, 65], which is a typical period of signs of deep SLE pathology of MRL-lpr/lpr mice [66]. Obviously, that the state of “health” in the case of autoimmune-prone mice should be considered quite conventional, the development autoimmune pathology nevertheless is spontaneous, and AI processes leading to deep pathology increase gradually. To distinguish different levels of SLE development, MRL-lpr/lpr mice without typical autoimmune symptoms and demonstrating no abzyme activities (similar to nonautoimmune healthy control mice) were independently of age tentatively designated as healthy mice, while the animals having no visual or biochemical SLE symptoms but demonstrating detectable activities of abzymes were provisionally named as prediseased MRL-lpr/lpr mice. Mice demonstrating all visual symptoms and biochemical indexes of SLE were designated as diseased animals. We have compared healthy (2–3 months of age) and spontaneously diseased MRL-lpr/lpr mice with all visible symptoms no older than 7 months [29–31]. For a more precise characterization of the various states of these mice, we have evaluated a variety of medical, biochemical, and immunological characteristics of their status including the relative levels of Abs to various autoantigens of abzymes demonstrating different catalytic activities.
\nThe average anti-DNA Abs concentration in the case of (CBAxC57BL)F1 and BALB/c nonautoimmune control mice was estimated to be approximately 0.03–0.04 A450 units and was comparable with that for healthy autoimmune-prone MRL-lpr/lpr mice (0.032 A450 units) [29–31]. After spontaneous development of SLE in MRL-lpr/lpr mice (depending of individual mice during 4–7 months), it increases to 0.2 A450 units, but there were no remarkable change in the anti-DNA Abs titers in the case of control nonautoimmune mice during 7–8 months of the experiment [29–31]. After MRL-lpr/lpr mice immunization with complex of methylated-BSA with DNA (further marked as DNA), the average concentration of anti-DNA Abs increased to approximately 0.6 A450 units [29–31]. It should be mentioned that IgG antibodies from the sera of control 2–7 month-old nonautoimmune CBA and BALB mice and conditionally healthy 2–3 months old MRL-lpr/lpr mice were shown to be catalytically inactive [29–31]. At the same time, during spontaneous development of SLE and especially after MRL-lpr/lpr mice immunization with DNA the relative catalytic DNase activity was significantly increased. Figure 1(A) demonstrates hydrolysis of supercoiled (sc) plasmid DNA by IgGs from various mice after 2 h of incubation. To quantify the DNase activity, a concentration of each electrophoretically and immunologically homogeneous IgG preparation (containing no any canonical enzymes) converting scDNA to relaxed DNA during 0.2–4 h of incubation without formation of linear or fragmented DNA was used (for example, lanes 1–3, Figure 1A). The relative efficiency of DNA hydrolysis was estimated from the relative percentage of DNA in the band of sc and relaxed DNA; the relative amount of DNA in these two bands for DNA incubated without IgGs or with Abs from healthy mice was taken into account. The measured relative activities (RAs) were normalized to standard conditions (0.1 mg/ml Abs, 2 h) and a complete hydrolysis of scDNA giving hydrolyzed form was taken for 100% of DNase activity. The RAs of IgGs in the hydrolysis of ATP (Figure 1B) and maltoheptaose (MHO; Figure 1C) were also estimated using the same approach as in the case of DNase activity.
\nDetermination of relative DNase (A), ATPase (B), and amylase (C) activities of catalytic IgGs (0.1 mg/ml) from different mice [
All data obtained are given in Table 1. One can see, that at 7 months of age before development of visible pathology markers (similar to mice of 1–3 months of age before deep MRL-lpr/lpr mice spontaneous pathology) MRL-lpr/lpr mice demonstrate no proteinuria (urine proteins <3.0 mg/ml). In addition, they are characterized by a relatively weak increase in the concentrations of Abs to native and denatured DNA. Moreover, the values of these parameters for some individual prediseased mice are comparable with the values observed for healthy mice. Interestingly, IgGs from sera of healthy MRL-lpr/lpr and control CBA, BALB mice possess well-determined amylase activity. This activity increased in the case of prediseased MRL-lpr/lpr mice, but the observed difference with healthy animals was not statistically significant. The changes in this parameter become statistically significant only for mice with deep pathology (Table 1). It is necessary to emphasize that IgGs of healthy MRL-lpr/lpr mice do not possess any DNase and ATPase activities and only the increase in these activities at a predisease stage should be considered as statistically significant indicator of the outset of spontaneous autoimmune disease of mice. After developing of deep SLE pathology, the RAs of DNase activity in the case of male and female mice increases 3.3- and 7.3-fold, respectively, comparing to prediseased mice, while increase in ATPase activity is significantly greater, 27- and 171-fold, respectively (Table 1). Thus, only these activities are the most important indicators of predisease state and deep pathology of MRL-lpr/lpr mice.
\nGroup description | \nNumber of mice | \nUrine protein, mg/ml** | \nAbs to native DNA, A450* | \nAbs to denatured DNA, A450* | \nDNase activity, %* | \nATPase activity, %* | \nAmylase activity, %* |
---|---|---|---|---|---|---|---|
Control males and females | |||||||
(CBA × C57BL) F1 (3–7 mo.) | \n8 (4 f + 4 m) | \n0.12 ± 0.07 | \n0.04 ± 0.01 | \n0.02 ± 0.01 | \n0*** | \n0*** | \n1.0 ± 0.5*** |
BALB/c (3–7 mo.) | \n8 (4 f + 4 m) | \n0.1 ± 0.08 | \n0.03 ± 0.01 | \n0.017 ± 0.004 | \n0 | \n0 | \n1.1 ± 0.5 |
\n | MRL-lpr/lpr males | ||||||
Healthy (2–3 mo.) | \n5 | \n0.38 ± 0.02 | \n0.032 ± 0.01 | \n0.09 ± 0.07 | \n0 | \n0 | \n1.9 ± 1.2 |
Healthy, pre-diseased (7 mo.) | \n5 | \n0.8 ± 0.3 | \n0.11 ± 0.02 | \n0.16 ± 0.05 | \n3.1 ± 1.4 | ||
Diseased (7 mo) | \n8 | \n8.0 ± 3.1 | \n0.18 ± 0 .08 | \n0.23 ± 0.11 | \n|||
Immunized | \n6 | \n9.5 ± 1.7 | \n0.6 ± 0.17 | \n1.1 ± 0.16 | \n|||
\n | MRL-lpr/lpr females | ||||||
Healthy (2–3 mo) | \n5 | \n0.31 ± 0.03 | \n0.08 ± 0.03 | \n0.12 ± 0.06 | \n0 | \n0 | \n1.8 ± 1.1 |
Healthy, pre-diseased (7 mo) | \n5 | \n0.9 ± 0.2 | \n0.20 ± 0.06 | \n0.08 ± 0.04 | \n3.6 ± 1.4 | ||
Diseased (7 mo) | \n5 | \n5.0 ± 3.8 | \n0.16 ± 0.12 | \n0.21 ± 0.12 | \n
Autoimmune characteristics of AI-prone MRL-lpr/lpr and control non-autoimmune mice [31].
For each mouse, the mean of three repeats is used.
Proteinuria corresponds to ≥3 mg of total protein/ml of urine.
100% relative activity corresponds to a complete transition of the substrate to its products after the hydrolysis in the presence of 0.1 mg/ml IgGs.
Statistically significant changes in parameters are given in bold.
The relationship between the relative activities of abzymes and the formation of following hematopoietic progenitors colonies has been analyzed: CFU-GM, granulocytic-macrophagic colony-forming unit; CFU-E, erythroid burst-forming unit (late erythroid colonies); CFU-GEMM, granulocytic-erythroid-megacaryocytic-macrophagic colony-forming unit; and BFU-E, erythroid burst-forming unit (early erythroid colonies) [29–31]. In the bone marrow of healthy MRL-lpr/lpr males and females (3 months of age), normal distribution of committed progenitors was observed, and the blood serum IgGs in these mice as well as control CBA mice show no detectable DNase and ATPase activities (Table 2). In MRL-lpr/lpr males and females (7 months old) having no proteinuria and SLE clinical manifestations but demonstrating detectable activities of abzymes, the relative number of BFU-E and CFU-GEMM colonies increased ~2- and ~16.4–28.4-fold, respectively. For spontaneously deep diseased males and females showing high RAs of DNase and ATPase activities, the profile of HSC differentiation was changed significantly comparing with prediseased mice: BFU-E colonies number increased approximately two times, while the number of CFU-GEMM and CFU-GM colonies decreased by factors of 2.4–3.4 and ~2.6–4.0, respectively. After the development of SLE induced by the mice immunization with DNA, the highest rise in anti-DNA Abs, Abz activities, and proteinuria was observed [29–31]. In addition, a very specific differentiation profile of HSC was revealed (Table 2). The numbers of CFU-GEMM and BFU-E colonies were 4.3- and 3.6-fold lower than for spontaneously diseased mice, while the number of CFU-GM colonies was comparable. Interestingly, the profiles of bone marrow HSC differentiation for immunized mice and healthy mice were not much different (Table 2). The data of Tables 1 and 2 are summarized in Figure 2.
\nGroup description | \nVisual symptoms | \nNumber of mice | \nNumber of colonies* | ||
---|---|---|---|---|---|
BFU-E | \nCFU-GM | \nCFU-GEMM | |||
CBA (3–7 mo) | \nNo | \n8 | \n3.0 ± 0.5** | \n7.3 ± 1.0** | \n0.25 ± 0.05** |
\n | MRL-lpr/lpr males | ||||
Healthy (2–3 mo.) | \nNo | \n5 | \n6.5 ± 1.5 | \n7.0 ± 1.0 | \n0.5 ± 0.1 |
Healthy, pre-diseased (7 mo) | \nNo | \n5 | \n12.7 ± 1.4 | \n30.0 ± 1.3 | \n9.2 ± 1.9 |
Diseased (7 mo) | \nYes | \n5 | \n25.3 ± 9.8 | \n7.4 ± 0.4 | \n3.9 ± 2.0 |
Immunized (3 mo) | \nyes, weak | \n5 | \n7.0 ± 2.1 | \n6.0 ± 2.6 | \n0.9 ± 0.7 |
\n | MRL-lpr/lpr females | ||||
Healthy (2–3 mo) | \nNo | \n5 | \n5.5 ± 0.5 | \n11 ± 2.5 | \n0.5 ± 0.2 |
Healthy, pre-diseased (7 mo) | \nNo | \n5 | \n11.5 ± 2.0 | \n23.0 ± 3.0 | \n8.2 ± 3.0 |
Diseased (7 mo) | \nYes | \n5 | \n22.1 ± 8.0 | \n9.0 ± 3.9 | \n2.4 ± 1.8 |
Formation of bone marrow progenitor colonies in from control nonautoimmune and MRL-lpr/lpr mice [31].
For each mouse, the mean of four repeats is used.
Mean ± confidence interval.
The relative profile of differentiation of bone marrow progenitors. Relative number of total erythroid cells (BFU-E+ CFU-E), CFU-GM, and CFU-GEMM colonies in the case of healthy CBA, conditionally healthy MRL-lpr/lpr mice at 3 months of age, after MRL-lpr/lpr mice development of pre-disease and deep SLE, as well as after mice immunization with DNA is shown [
One can see that in the condition of predisease a strong increase in the relative number of CFU-GM colonies is observed, and at this time, there is a reliable and statistically significant appearance of DNase and ATPase activities of IgG antibodies. At transition from predisease condition to deep SLE pathology, there is additional change in profile differentiation of bone marrow HSC; the number of CFU-GM colonies is significantly decreased, but at the same time, a significant increase in BFU-E cells is observed. Such change in differentiation profile of bone marrow HSC is associated with significant increase in DNase and ATPase activities of IgGs. Healthy MRL-lpr/lpr mice 3 months of age treated with DNA show a very strong increase in DNase and ATPase activities, but differentiation profile of bone marrow HSC is almost same as for healthy male and female MRL-lpr/lpr mice. This could indicate that appearance of abzymes in healthy mice after their immunization with DNA is not associated with the change of differentiation profile of bone marrow HSC, and there may be some other ways of this phenomenon realization. Taking this into account, we analyzed lymphocyte proliferation in different organs of MRL-lpr/lpr mice [29–31].
\nSpontaneous development of SLE results in remarkable average increase in lymphocyte proliferation in all analyzed organs of males and females comparing with healthy control mice (Figure 3) [29–31].
\nThe relative level of lymphocyte proliferation in different organs of male healthy CBA, conditionally healthy MRL-lpr/lpr mice at 3 months of age, after development of pre-disease and deep SLE pathology MRL-lpr/lpr in mice, as well as after mice immunization with DNA is shown [
Interestingly, the relative level of lymphocyte proliferation in the spleen of healthy control CBA mice is approximately threefold higher than that for healthy MRL-lpr/lpr mice (Figure 3). Transition from healthy to spontaneously prediseased mice leads to the increase in lymphocyte proliferation in spleen by ~2.8-fold in parallel with increase of average level of the proliferation 1.4- and 1.8-fold in lymph nodes and thymus, respectively [29–31]. While there is no remarkable difference in lymphocyte proliferation in bone marrow of healthy CBA, healthy and prediseased MRL-lpr/lpr mice, the diseased animals demonstrate increase in this parameter by a factor of ~1.5 (Figure 3). The spontaneous pathology of MRL-lpr/lpr mice develops slowly and most of the mice are showing signs of the deep disease only from 5 to 9 months of life. The average values of the lymphocyte proliferation of diseased MRL-lpr/lpr mice are significantly increased in all organs compared to healthy mice. Interestingly, immunization of healthy 3 months old MRL-lpr/lpr mice with DNA leads to the increase of lymphocyte proliferation in all organs except thymus at 20 days after their treatment (Figure 3). Thus, a significant increase in the relative level of lymphocyte proliferation in mice may be an important factor causing the development of abzymes with very high activity after their immunization with DNA. As mentioned above, mice immunized with DNA do not show significant changes in bone marrow HSC differentiation profile. However, DNA-treated mice demonstrate high level of bone marrow lymphocyte proliferation (Figure 3). This may be due to the fact that DNA has little effect on the profile of stem cells differentiation but effectively stimulates increase in the proliferation and changes the profile of mouse bone marrow lymphocyte differentiation.
\nIn this regard, data comparing relative Abz activities from serum and cerebrospinal fluid (CSF) of the same MS patients with multiple sclerosis should be noted [67–69]. It was shown that IgGs from sera and cerebrospinal fluid of MS patients are active in the hydrolysis of DNA, MBP, and oligosaccharides. In addition, the specific RAs of these abzymes from the CSF of MS patients are dependently on their different activities were approximately 30- to 60-fold higher comparing to serum Abzs from the same patients [67–69]. It means that during spontaneous or induced development of ADs as a result of specific differentiation of lymphocytes in cerebrospinal fluid there may be formation of cells producing different catalytic antibodies directly in cerebrospinal fluid. Thus, one cannot exclude that the increased level of Abz activities in MRL-lpr/lpr mice immunized with DNA may be a consequence of specific additional differentiation of naive lymphocytes not only in different organs but also in the cerebrospinal fluid. Another important factor reducing the relative number of lymphocytes producing abzyme may be cell apoptosis.
\nIt is known that in norm (healthy mammals) harmful cells including lymphocytes are eliminated by apoptosis [61, 62]. The decrease in the lymphocyte apoptosis, producing Abzs harmful to mammals, can lead to increase in autoimmune reactions and acceleration of ADs development. The relative level of lymphocyte apoptosis in different organs and tissues of MRL-lpr/lpr mice was analyzed (Figure 4) [31].
\nThe relative level of lymphocyte apoptosis in different organs of male healthy CBA, conditionally healthy MRL-lpr/lpr mice at 3 months of age, after MRL-lpr/lpr mice development of pre-disease, and deep SLE pathology, as well as after mice immunization with DNA [
In control CBA and healthy MRL-lpr/lpr mice, the cell apoptosis level in different organs on average was comparable (Figure 4). The prediseased mice demonstrated relatively low decrease in lymphocyte apoptosis in bone marrow, but its remarkable increase in spleen. Transition from predisease state to deep SLE led to a significant decrease in the cell apoptosis level in all organs comparing with healthy and prediseased mice and maximal decrease was observed for bone marrow lymphocytes (Figure 4). However, the statistically significant two- to threefold maximal decrease in the apoptosis level was observed for bone marrow, thymus lymph nodes, and spleen of the mice immunized with DNA, which correlates with a very strong increase in the specific Abz activities of treated mice comparing to the spontaneously diseased animals (Figure 4). Therefore, it should be assumed that in the case of mice predisposed to ADs, introduction of foreign antigen can inhibit the elimination of harmful lymphocytes including ones producing dangerous abzymes by apoptosis and stimulate the proliferation of such cells. Overall, immunization of healthy MRL-lpr/lpr mice with DNA leads to the production of abzymes with very high activities, but it is not associated with noticeable change in profile of HSC differentiation but mainly caused by increase in lymphocyte proliferation and specific suppression of lymphocyte apoptosis in different organs [35]. In this regard, it should be mentioned that these regularities may to some extent be common in the development of various autoimmune diseases. For example, using experimental C57BL/6 autoimmune encephalomyelitis (EAE) mice (a model mimicking human MS), it was recently shown that spontaneous EAE development leads to the production of Abs to myelin basic protein (MBP) and DNA and to Abzs efficiently hydrolyzing these substrates, which associated with significant changes of the differentiation profile and level of lymphocytes proliferation of mice bone marrow HSC [32]. Immunization of these mice with MOG35 results in a very strong acceleration in the development of EAE and a very strong increase of the relative activities of abzymes hydrolyzing MOG, MBP, and DNA. The relative percent contents of total erythroid cells, CFU-GEMM and CFU-GM colonies at 3 months of age in healthy, nonautoimmune CBA, conditionally healthy, MRL-lpr/lpr and EAE C57BL/6 mice before and after development of these autoimmune pathologies were compared (Figure 5).
\nThe relative content (%) of total erythroid cells (BFU-E+ CFU-E), CFU-GM, and CFU-GEMM colonies in the case of healthy CBA, conditionally healthy MRL-lpr/lpr and C57BL/6 mice at 3 months of age, and after development of, respectively, EAE and SLE is shown [
One can see that the relative content (percent of total number of different cells) of various bone marrow colonies after spontaneous achievement of deep pathologies by MRL-lpr/lpr and EAE mice is nearly the same. In addition, the level of lymphocyte proliferation as well as cell apoptosis in different organs including bone marrow of healthy MRL-lpr/lpr and EAE mice, at stages of their prediseases and deep pathologies were also comparable [32]. In addition, it was shown that abzymes hydrolyzing MBP are formed in the early stages of human MS, unlike in later stages of SLE, while the reverse situation was observed for abzymes with DNase activity [70–87]. One gets the impression that SLE and MS differ greatly in their initial stages but become to some extent more similar at the later stages of these diseases. The blood of patients with SLE and MS after a long illness contains Abs to a variety of autoantigens and abzymes hydrolyzing nucleotides, oligosaccharides, lipids, DNA, RNA, MBP, and many other proteins [13–21, 70–93].
\nClinically definite MS diagnosis is more often based on tomographic detection of brain-specific plaques appearing on late stages of this disease. But, similar brain plaques were also detected on the late stages of SLE [38, 41]. MS is a central nervous system disease resulting in the manifestation of different psychiatric and nervous disturbances. Neuropsychiatric disturbances occur also in about 50% of patients with SLE and carries a poor prognosis (reviewed in [41]). SLE affects mainly on the central nervous system and it supposedly more than any other inflammatory systemic disease causes various psychiatric disorders [41]. Peripheral nervous system involvement seems to be much less. Neural cell injury and rheological disturbances mediated by auto-Abs may be due to two of the main possible mechanism of tissue damage [41]. Interplay between these processes is determined by genetic factors, and may be modulated by hormones, complicated by a many of secondary factors, may explain the wide spectrum of features revealed in SLE [41]. Thus, not only specific changes in the profile of differentiation of bone marrow stem cells, increased levels of lymphocyte proliferation and suppression of apoptosis of harmful cells in different organs leading to the production of dangerous abzymes with various activities, but also some other indicators of different psychiatric and nervous disturbances in varying degrees, are common for some patients with SLE and MS.
\nThe sera of healthy humans and mammals contain usually autoantibodies to many different antigens, including RNA, DNA, proteins, and other antigens [13–22, 43, 44]. Natural abzymes from the sera of patients with different AI diseases are products of different immuno-competent cells and usually polyclonal in origin ([13–22] and refs cited here). Purification of natural abzymes containing no canonical enzymes is a very important task in their study; peculiarities of such antibodies isolation were discussed in detail in reviews [13, 19]. Electrophoretically and immunologically homogeneous IgG fractions with or without different catalytic activities from the sera of healthy donors and autoimmune patients described in this chapter were first purified using Protein G-Sepharose, while IgAs and IgMs by affinity chromatography on Protein A-Sepharose under conditions removing nonspecifically bound proteins. Then IgAs were separated from IgMs and IgGs from possible admixtures of canonical enzymes by FPLC gel filtration in acidic conditions (pH 2.6) destroying immuno-complexes [29–32, 49, 50, 55–57, 64–103]. Overall, ~900 kDa IgMs, 170 kDa IgAs, and 150 kDa IgGs did not contain possible contaminating proteins detected by acrylamide gel silver staining under reducing and nonreducing conditions.
\nThe application of rigid criteria allowed the authors of the first article describing natural abzymes hydrolyzing vasoactive intestinal peptide to obtain irrefutable evidence that this activity is an intrinsic property of IgGs from sera of patients with asthma [9]. Later several additional rigid criteria were proposed (reviewed in Refs. [13, 19]). We applied a set of these strict criteria [9, 13–22] for the analysis of DNase and RNase [11, 31, 64, 65, 70–75], MBP-hydrolyzing [76–79], ATPase [30], and amylase [29, 88–91] activities as intrinsic properties of IgG and/or IgM and IgA antibodies from sera of SLE patients and mice. Several more important of them may be summarized as follows: (1) all Abs were electrophoretically homogeneous; (2) FPLC gel filtration of these Abs under conditions destroying strong noncovalent complexes (acidic buffer, pH 2.6) did not abolish these activities, and activities peaks exactly coincided with peaks of intact Abs; (3) immobilized mouse IgGs against the light chains of human Abs absorbed completely these activities; these activity’s peaks coincided with the peaks of Abs eluted using acidic buffer; and (4) F(ab) and F(ab)2 fragments of Abs showed to some extent comparable levels of the activities comparing with intact Abzs. To exclude possible artifacts causing by hypothetical traces of canonical enzymes, Abs from sera of SLE patients were subjected to SDS-PAGE in a gel copolymerized with polymeric DNA or RNA and their nuclease activities were detected
Analysis of the implementation of strict criteria of intrinsic enzymatic activities of IgGs from sera of SLE patients and mice. SDS-PAGE analysis of homogeneity of IgGmix (mixture of equal amounts of Abs from sera of 10 SLE patients) before (lane 2) and after (lane 3) Abs boiling with DTT; lane 1 shows positions of protein markers (A) [
Application of the strict criteria to show that MBP-hydrolyzing activity is intrinsic property of IgGs from sera of SLE patients [
Similar rigid criteria were used by us for evidence of the catalytic activity belonging to the antibodies, hydrolyzing nucleotides, DNA, RNA, various peptides, proteins, oligosaccharides from blood with different autoimmune diseases, and animals immunized with different antigens [11, 13–22, 32, 49–57, 64–65, 67–103].
\nHealthy humans do not demonstrate antibodies with detectable DNase and RNase activities, their levels are more often on the borderline of sensitivity of the detection methods [13–22]. The RAs of DNase and RNase abzymes from the sera of patients with SLE vary markedly from patient to patient [13–22].
\nWe analyzed the possible heterogeneity of catalytic properties of polyclonal DNase and RNase IgGs from SLE patients and observed an extreme heterogeneity in kinetic and thermodynamic parameters, relative specific activities and substrate specificities, which are different very much from patient to patient. Chromatography on DNA-cellulose showed that only 10–30% of the total electrophoretically homogeneous IgGs and IgMs dependently on patient may be bound to the affinity sorbent. Interestingly, when Abs were eluted from DNA-cellulose by a NaCl gradient (0–3 M) and then acidic buffer (pH 2.6) the Abs and their DNase and RNase activities were distributed all over the chromatography profile (for example, Figure 8A) [70, 71]. The same situation was observed for Abs with nuclease activities from sera of MS patients [8, 80–82], and rabbits immunized with DNA, RNA, DNase I, DNase II, and pancreatic RNase [49, 50, 55–57]. The affinity of Abs fractions for these substrates was increased gradually with the increase in eluting salts concentrations. When IgGs eluted from DNA-cellulose were fractionated on Sepharose bearing immobilized monoclonal mouse Abs against anti-kappa or anti-lambda human IgGs, 60–70% of IgGs were adsorbed by Abs against lambda- and 30–40% by Abs against kappa-Abs (Figure 8B) [70, 71].
\nSeparation of SLE IgMs having affinity to DNA by affinity chromatography on DNA-cellulose (A) [
The fractions corresponding IgGs with kappa-light chain were about 30- to 50-fold more active in hydrolysis of both RNA and DNA than lambda-IgGs. SLE IgGs and IgAs with DNase activity [70, 71] similarly to Abs with nuclease activity from sera of patients (and mice) with other autoimmune diseases [80–82, 95, 99–103] efficiently hydrolyzed all single- and double-stranded DNA of different sequences and length. The substrate specificity of SLE IgGs with RNase activity, however, was unique within certain limits for Abs from every individual SLE patient [11]. In contrast to human RNases, SLE IgGs effectively hydrolyze the most resistant poly(A) substrate for all known human RNases [104–109]. SLE IgGs demonstrated a very slow hydrolysis of poly(C) [11], which is the best substrate for all mammalian RNases [104–106]. Therefore, for more detail analysis of IgGs of SLE patients ribo(pA)13 was used. Figure 8(C) demonstrates pH dependence of [5′-32P](pA)13 hydrolysis by three of six SLE IgGs analyzed and by two human blood RNases. All six dependences showed individual features of pH dependencies. In contrast to all SLE abzymes, RNases have only one pH optimum (6.8–7.5) for hydrolysis of poly(A) [104–106] (Figure 8D). Polyclonal SLE abzymes more often shows high activity at pH from 6.0 to 9.5. For example, preparation Abz-1 demonstrates maximal activity at pH 8.8; Abz-2 shows three marked pH optima at pH 8.5, 7.7, and 7.2, while Abz-5 hydrolyzes RNA with comparable efficiency at pH values from 6.0 up to 9.5 (Figure 8C). Abz-3 and Abz-4 also demonstrated several pronounced optima at pH 6.0–9.5 similar with that for Abz-2, while Abz-6 showed no optimum, like Abz-5.
\nInterestingly, even at fixed pH 7.5 initial rates corresponding to increase in oligonucleotide concentrations were consistent with Michaelis-Menten kinetics only for all human RNases and Abz-1, for which was only one interception of curves in coordinates of Cornish-Bowden (Figure 9A) [71]. Three IgGs (Abz-2–Abz-4) demonstrated several apparent values of both
Initial rates of [
Substrate | \nPreparation | \n||
---|---|---|---|
Antibodies | |||
d(pA)13 | \nAbz-1 | \n7 × 10−8 | \n2.0 × 10−2 |
(pA)13 | \n\n | 4 × 10−8 | \n1.4 |
d(pA)13 d(pA)13 | \nAbz-2 | \n4.7 × 10−8 to 3.0 × 10−7 | \n2.0 × 10−3 to 7.1 × 10−2 |
(pA)13 | \n\n | 5.1 × 10−8 to 4.4 × 10−7 | \n0.12–0.84 |
p(U)10 | \n\n | 9.0 x 10−8 – 4.1 x 10−7 | \n3.2 x 10−3 – 1.3 x 10−2 |
(pA)13 | \nAbz-3–Abz-4 | \n1 × 10−8 to 2 × 1 0−6 | \n1.0 × 10−2 to 2.5 |
RNases | |||
p(A)13 | \nRNase A | \n3.4 x 10−6 | \n2.2 x 10−2 |
p(A)13 | \nRNase 3 | \n4.9 × 10−6 | \n1.7 × 10−2 |
p(U)10 | \n\n | 2.1 × 10−6 | \n37 |
p(A)13 | \nRNase 4 | \n7.2 × 10−6 | \n5.2 × 10−2 |
p(U)10 | \n\n | 5.6 × 10−6 | \n26 |
Kinetic parameters for hydrolysis of different oligonucleotides by catalytic SLE-IgGs, human serum RNases and pancreatic ribonuclease A [71].
The errors of the values determination were within ± 10–30%.
It should be mentioned that the same preparations of polyclonal Abzs hydrolyzed RNA approximately 10- to 300-fold faster than DNA [70, 71]. In addition, several monoclonal IgGs against B-DNA of different sequences (from SLE mice) efficiently hydrolyze single- and double-stranded RNA and DNA in a sequence-independent manner, the RNase activity was by a factor of 30–100 higher than of DNA [107]. Our findings indicate that a variety of anti-DNA and anti-RNA abzymes are able to hydrolyze both DNA and RNA [13–22]. In this respect, it should be mentioned that after immunization of rabbits with DNA, RNA, DNase I, DNase II, and pancreatic RNase I, antinuclease IgGs with different affinity to DNA were separated for several fractions by chromatography on DNA-cellulose [49, 50, 55–57]. IgGs of all fractions demonstrated DNase and RNase activity and RNase activity was 10- to 50-fold higher than DNase one. Only one small fraction in the case of abzymes obtained after immunization of rabbits with RNA demonstrated only DNase activity and was not able to hydrolyze RNA [50]. The data obtained testify in favor of the formation of abzymes with chimeric structure of the active centers, which are mostly able to hydrolyze both DNA and RNA.
\nCanonical RNases are usually specific for sequences (for example, RNase T1 is specific for guanosines, while RNase A for Py-A sequences) or for structural features (nuclease S1, for example, hydrolyzes only single-stranded domains of RNA). Abzymes of SLE and other autoimmune patients demonstrate novel RNase activities. Some of them may be stimulated by Mg2+; they are not sequence-specific but sensitive to subtle and/or drastic folding changes of structurally well-characterized tRNA [72–75]. Two tRNALys [72, 74], one corresponding to human mitochondria, while the second tRNALys is a mutant revealed in patients with myoclonic epilepsy, in which A nucleotide at position 50 is changed for G nucleotide. Different canonical RNases including RNase A showed no difference in cleavage patterns of these tRNALys [72]. However, in the presence of Mg2+ RNase SLE abzymes produced new cleavage sites; the mutant tRNALys showed a significantly different sensitivity for abzymes in the substrate mutated region, and the hydrolysis was detected at new positions, showing local structural or conformational changes of tRNALys.
\nMost of Mg2+-dependent abzymes display usually no sequence specificity; they are more sensitive to structural features of different tRNAs specific for Gln, Phe, Asp, and Lys [72–75]. Abzymes of some AI patients demonstrate RNase A-type as a major specificity showing minor differences (preference for CpA and UpA sequences). However, some abzymes contain a major subfraction demonstrating T1-type of RNase specificity. The Mg2+-stimulated RNase IgGs had more often a cobra venom RNase V1-like specificity in cleavage of tRNAPhe with a unique Mg2+-dependent specificity toward double-stranded regions [72, 74]. In spite of some similarities, SLE abzymes show specificities quite different from that of RNase V1, but this specificity remarkably differs from patient to patient. Overall, monoclonal SLE abzymes entering to total pool of Abzs can discriminate between subtle or large structural changes and nucleotide sequences. Interesting that IgGs from patients with different AI and viral diseases can demonstrate different patterns of various tRNAs cleavage [72–75].
\nAbzs from the sera of patients with MS [80–82], viral hepatitis [100], HIV-infected patients [95], with tick-borne encephalitis [101], Hashimoto’s thyroiditis [102], and schizophrenia [103] also demonstrated DNase activity. All these diseases are characterized by different levels of the relative activity of abzymes, and DNase activity increases approximately in the following order: diabetes ≤ viral hepatitis ≈ tick borne encephalitis < polyarthritis ≤ Hashimoto’s thyroiditis ≤ schizophrenia < AIDS ≤ MS < SLE [13–22, 80–82, 95, 101–103]. Overall, DNase and RNase polyclonal Abzs from sera of patient with SLE, MS, and other AI and several viral diseases may be characterized by a relatively small or extremely large content of polyclonal nuclease abzymes containing different relative amounts of kappa- and lambda-Abzs, demonstrating from one to several pH optima, having a different net charge, may be dependent or not on different metal ions, and demonstrate different substrate specificities.
\nThe active centres of DNase, RNase, protease, and oligosaccharide-hydrolyzing abzymes from SLE, MS, and patients with other diseases are usually located on the light chains of these Abs [13–22]. The heavy chain is mainly responsible for specific antigen recognition and significantly increased antigen affinity for antibodies. The isolated light chains of IgGs cleavage vasoactive intestinal peptide with the activity 32-fold higher than Fab fragments [9]. Isolated by SDS-PAGE light chains of different abzymes were more active than the intact ones [13–22]. But, only multiple myeloma Bence-Jones proteins should be considered as natural human monoclonal abzymes [60].
\nA phagemid library of immunoglobulin kappa light chains derived from lymphocytes of peripheral blood of three SLE patients (106 variants) was cloned in pCANTAB5His6 vector. For amplification of the phage library
Affinity chromatography of phage particles on DNA-cellulose: (—) and (− − −), A280 values correspond respectively to the material of phage particles containing or not cDNA of kappa light chains [
The phage particles were distributed between 16 fractions of 11 peaks and all fractions corresponding to new small pools of anti-DNA MLChs demonstrated DNase activity (Figure 10). Thus, all small pools of MLChs of 16 fractions with different affinity of phage particles for DNA contained not only light chains without, but also with DNase activity. For preparation of individual colonies,
MLChs of 15 single colonies were used for purification of individual MLChs using chromatography on Ni2+-charged HiTrap chelating Sepharose and by following FPLC gel filtration [111]. The preparations of ~28-kDa MLChs were electrophoretically homogeneous, showed positive answer with mouse IgGs against human Abs light chains at Western blotting and positive ELISA answer using plates containing immobilized DNA;
The dependences of DNase RAs for various MLCh preparations on the concentration of different metal ions were analyzed. It was shown that MgCl2 and MnCl2 are good activators of all 15 MLChs. Since K+ and Na+ ions can influence on the spatial structures of different enzymes, antibodies, and nucleic acids, we have analyzed dependencies of the RAs upon concentration of these ions at fixed concentrations of MgCl2 and MnCl2 (2 mM). All MLChs demonstrated very specific dependencies and optimal concentrations of NaCl and KCl (for example, Figure 11). Optimal concentrations of NaCl and KCl for 15 MLChs are given in Table 4. It was interesting to see whether optimal concentrations of MgCl2 and MnCl2 can depend on NaCl and KCl concentrations. Figure 12 shows that for all MLChs in the presence of KCl and NaCl in different concentrations the dependencies reach plateau at 1.5–2.0 mM concentration of MgCl2 and MnCl2. Several MLChs demonstrated shape-bell dependencies; but the inhibition was usually observed at Mn2+ and Mg2+ concentrations higher than 2–4 mM (Figure 12) [111].
\nNumber of MLCh preparation | \nMnCl2 | \nMgCl2 | ||
---|---|---|---|---|
[KCl], mM | \n[NaCl], mM | \n[KCl], mM | \n[NaCl], mM | |
1 | \n0–0.5; > 0.5 inhib.a | \n0–0.5; > 0.5 inhib. | \n30.0; > plateaub | \n100.0; >120.0 inhib.a |
2 | \n100; >120 inhib. | \n100; > 120 inhib. | \n75.0; >100 inhib. | \n15.0; >100.0 inhib. |
3 | \n5.0; > 10 inhib. | \n5.0; > 10.0 inhib. | \n0.0; > 0.0 inhib. | \n0.0; > 0.0 inhib. |
4 | \n2.5; > 4.0 inhib. | \n7.5; > 10.0 inhib. | \n5.0 > 10.0 inhib. | \n8.5; > 15.0 inhib. |
5 | \n5.0; > 10 inhib. | \n5.0; > 10.0 inhib. | \n0.0 > 0.0 inhib. | \n0.0; > 0.0 inhib. |
6 | \n1–2 and 75; >75 inhib.c | \n1–2 and 150 | \n1.5–30 plateau; >40.0 inhib. | \n1–3 and 50.0; >100.0 inhib. |
7 | \n0.5; >1.0 inhib. | \n2.5; >3.0 inhib. | \n3.0; >4.0 inhib. | \n8.0; >10.0 inhib. |
8 | \n0–10 inhib.; 75; >80 inhib. | \n5–150 plateau | \n5.0 and 75.0; >80.0 inhib. | \n1.5; 2.0–75 inhib.; 100; >plateauc |
9 | \n20–25; >30 inhib. | \n2.0; >3.0 inhib. | \n10.0; >12.0 inhib. | \n5.0 and 75.0; >80.0 inhib. |
10 | \n2.5; plateaub | \n10.0; >15.0 inhib. | \n2.5; >5.0 inhib. | \n5.0; >7.0 inhib. |
11 | \n1–10 plateau; >10 inhib. | \n0.5–2.0 plateau; >2.0 inhib. | \n0–10.0 plateau; >10.0 inhib. | \n2–50 plateau; >60.0 inhib. |
12 | \n0.0; >0.0 inhib. | \n2.0 and 5–10; >10.0 inhib. | \n1–5; >10.0 inhib. | \n1–2 and 5–10; >10.0 inhib. |
13 | \n0.0; >0.0 inhib. | \n0.0 and 10.0; >10.0 inhib. | \n0.1–0.3; >0.7 inhib. | \n0–0.1; >0.1 inhib. |
14 | \n100–150 plateau | \n100–150 plateau | \n75.0; >100 inhib. | \n150.0 |
15 | \n1.5; >2 inhib. | \n50.0; >50.0 inhib. | \n0.0 and 50.0; >50.0 inhib. | \n0–10 and 50.0; >50.0 inhib. |
The optimal concentrations of KCl and NaCl in the case of individual recombinant MLChs in the presence of 2 mM MgCl2 or 2 mM MnCl2 [111].
For each value, a mean of two measurements is reported, optimal concentrations are given in bold; the mark (>value inhib.) means that the dependence demonstrates bell-shaped character and that at higher concentrations of the salt the inhibition of the reaction is observed.
The mark (30 (or any other value in bold); >plateau) means that optimal concentration corresponds to 30 mM and there is no remarkable inhibition up to maximal concentration (100–150 mM) of NaCl or KCl used.
The mark (1.5; 2.0–75 inhib.; 100; >plateau (or other similar values)) means that there are two optimal concentrations at 1.5 and 100 mM salt and a significant decrease in the activity of analyzed MLCh at concentrations in the region 2.0–75 mM.
Dependences of the RAs of DNase activity for several MLChs upon the concentration of KCl and NaCl at 2 mM concentrations of MgCl2 and MnCl2 (A–F) [
Dependences of the RAs of DNase activity for several MLChs (20 nM) on the concentrations of MnCl2 and MgCl2 at different fixed KCl and NaCl concentrations (A–D) [
Various canonical DNases demonstrate usually different pH optima, but all of them have only one pH optimum [108, 109]. In contrast to known canonical DNases, polyclonal DNase Abzs from the sera of patients with SLE and other diseases can contain from one to many monoclonal abzymes demonstrating from 1 to 2–8 well pronounced pH optima in range from 5 to 10 [13–22, 72, 73, 82–84, 110]. pH optima of 15 MLChs in the presence of 2 mM MgCl2 as well as for DNase II and DNase I were analyzed and several typical dependencies are given in Figure 13. DNase I has only one optimum at pH 7.0–7.2, while DNase II demonstrates one optimum at pH 4.9–5.0 (Figure 13A). Optimal pH for all 15 MLChs is given in Table 5 [111].
\nDNase or number of MLCh preparation | \nOptimal pH | |
---|---|---|
\n | pH1 | \npH2 |
DNase I | \n7.0–7.2 | \nNo second optimum |
DNAse II | \n4.9–5.0 | \nNo second optimum |
1 | \n5.7–5.9 | \n7.9–8.1b |
2 | \n6.0–6.2 | \n8.2–8.3 |
3 | \n6.9–7.0 | \n8.2–8.5 |
4 | \n7.5–7.6 | \nNo second optimum |
5 | \n6.9–7.0 | \n8.2–8.5 |
6 | \n7.8–8.0 | \nNo second optimum |
7 | \n6.2–6.4 | \nNo second optimum |
8 | \n8.5–8.6 | \nNo second optimum |
9 | \n7.8–8.0 | \n8.9–9.1 |
10 | \n6.1–6.3 | \n8.5–8.7 |
11 | \n4.8–5.0 | \n8.6–8.7 |
12 | \n7.7–7.9 | \nNo second optimum |
13 | \n8.5–8.7 | \nNo second optimum |
14 | \n7.9–8.1 | \nNo second optimum |
15 | \n5.4–5.6 | \nNo second optimum |
Dependences of the RAs of DNA-hydrolyzing activity of human DNase I and DNase II (A) and of nine MLChs (B–D) on pH of reaction mixtures [
Eight of 15 MLChs demonstrated only one pH optimum, while seven preparations shows two different pH optima (Figure 13). It was shown that Mn2+, Co2+, Ni2+, and Ca2+ ions activate DNase I in significantly smaller degree than Mg2+ ions [108, 109]. The RAs of 15 MLChs in the presence of 6 various metal ions (2 mM) using optimal NaCl and KCl concentrations and optimal pH were estimated (Table 6). The maximal activity for various MLChs was observed in the presence of different MeCl2 salts, but in average the activity decreased in the following order: MnCl2 > CoCl2 > MgCl2 > NiCl2 ≈ CaCl2 (Table 6). For all 15 MLChs, apparent
Number of MLChs | \nMg2+ | \nMn2+ | \nZn2+ | \nNi2+ | \nCo2+ | \nCu2+ | \nCa2+ | \npHc | \n[NaCl] or [KCl], mMd | \n|
---|---|---|---|---|---|---|---|---|---|---|
1 | \n92.7 | \n100a | \n6.5 | \n10.7 | \n20.6 | \n0.0 | \n4.0 | \n5.8e | \n100.0 Na+ | \n0.6 ± 0.04 |
60.7 | \n100.0 | \n7.5 | \n11.5 | \n18.5 | \n0.0 | \n5.4 | \n8.0ξ | \n100.0 Na+ | \n||
2 | \n69.6 | \n100.0 | \n55.4 | \n83.8 | \n73.0 | \n0.0 | \n10.1 | \n6.1 | \n80.0 Na+ | \n0.1 ± 0.01 |
100.0 | \n92.3 | \n18.5 | \n95.6 | \n63.7 | \n3.0 | \n26.0 | \n8.2 | \n80.0 Na+ | \n||
3 | \n78.3 | \n98.5 | \n42.7 | \n56.2 | \n0.3 | \n9.0 | \n100.0 | \n6.9 | \n5.0 K+ | \n|
\n | 1.3 | \n100.0 | \n3.6 | \n11.9 | \n99.0 | \n0.0 | \n0.0 | \n8.4 | \n5.0 K+ | \n0.33 ± 0.03 |
4 | \n51.0 | \n100.0 | \n80.0 | \n56.0 | \n24.0 | \n5.0 | \n26.0 | \n7.5 | \n7.5 Na+ | \n0.5 ± 0.04 |
5 | \n78.3 | \n98.5 | \n42.7 | \n56.2 | \n0.0 | \n9.0 | \n100 | \n6.9 | \n5.0 K+ | \n0.3± 0.03 |
\n | 1.3 | \n100.0 | \n3.6 | \n11.9 | \n99.0 | \n0.0 | \n0.0 | \n8.4 | \n5.0 K+ | \n0.34 ± 0.03 |
6 | \n1.3 | \n100.0 | \n3.6 | \n11.9 | \n99.0 | \n0.0 | \n0.0 | \n7.9 | \n1.0 K+ | \n0.02 ± 0.003 |
7 | \n75 | \n70 | \n80 | \n85 | \n100.0 | \n5 | \n21 | \n6.3 | \n2.5 Na+ | \n0.3 ± 0.04 |
8 | \n58.5 | \n100.0 | \n18.3 | \n19.7 | \n0.0 | \n0.0 | \n16.2 | \n8.5 | \n5.0 Na+ | \n0.06 ± 0.07 |
9 | \n100.0 | \n85.2 | \n15.9 | \n0.0 | \n85.4 | \n0.0 | \n75 | \n7.8 | \n1.5 Na+ | \n0.7 ± 0.08 |
10 | \n25 | \n20 | \n15 | \n100.0 | \n17 | \n2 | \n39 | \n8.6 | \n10.0 Na+ | \n0.2 ± 0.02 |
11 | \n80 | \n85 | \n43 | \n24 | \n100.0 | \n0.0 | \n8 | \n8.6 | \n5.0 K+ | \n0.7 ± 0.06 |
12 | \n55.0 | \n88.5 | \n16.5 | \n56.3 | \n100.0 | \n1.2 | \n56.2 | \n7.8 | \n5.0 K+ | \n0.12 ± 0.01 |
13 | \n34.5 | \n25.0 | \n16.9 | \n83.3 | \n100.0 | \n4.8 | \n41.9 | \n8.6 | \n0.1 K+ | \n0.13 ± 0.01 |
14 | \n80.9 | \n75.2 | \n19.7 | \n100.0 | \n59.5 | \n6.3 | \n37.3 | \n8.0 | \n5.0 Na+ | \n0.11 ± 0.01 |
15 | \n100.0 | \n49.2 | \n55.7 | \n33.5 | \n85.8 | \n45.0 | \n70.0 | \n5.5 | \n1.5 Na+ | \n0.12 ± 0.01 |
The RAs of different recombinant MLChs in the presence of various metal ions (2 mM) at optimal pHs and concentration of KCl and NaCl [111].
The maximal RAs in the presence of one of seven metal ions used was taken as 100% and given in bold; the error of the values determination (two independent experiments) did not exceed 7–10%.
The apparent
Optimal pHs were used for every of MLCh preparation.
Optimal concentrations of KCl and NaCl in the case of different MLChs were used.
The RAs for several MLChs were found in the case of two different pH optima.
Our previous findings demonstrated that polyclonal abzymes from sera of patients with SLE, MS, and other autoimmune and/or viral diseases can contain many monoclonal DNase and RNase abzymes showing very different enzymatic properties [13–22]. At the same time, estimation of possible number of monoclonal abzymes in their total pools was very difficult, since they can have comparable or different affinity for DNA, significantly different optimal pHs, various
In our study DNase activity only for 45 of 451 single colonies corresponding to only one (eluted with 0.5 M NaCl) of 16 fractions with different affinity for DNA-cellulose was analyzed, while MLChs of all these fractions effectively hydrolyze DNA [111]. Fifteen of 45 individual MLChs (~33%) were active in the hydrolysis of DNA. Taking into account the fact that only 45 of 451 colonies were analyzed, it should be assumed that even this fraction contains much greater number of monoclonal abzymes with DNase activity. In this regard it should be mentioned the data of other article [112]. After separation of phage particles on DNA-cellulose, the fraction eluted by an acidic buffer (pH 2.6) was used for obtaining of MLChs (~28 kDa) with DNase activity. In this case, 33 of 687 individual colonies were chosen randomly for study of MLChs. Nineteen of 33 clones (58%) demonstrated DNase activity [112]. Detection of DNase activity
The increased level of antibodies to myelin basic protein and abzymes hydrolyzing MBP was revealed for the first time in the blood of patients with multiple sclerosis [83–87]. It was shown that Abs of healthy donors cannot hydrolyze MBP [13–22, 83–87]. The most widely accepted theory of multiple sclerosis pathogenesis assigns the major role in the destruction of myelin including MBP to the inflammation related to autoimmune reactions [43, 113–115]. Increased levels of Abs and oligoclonal IgGs in the cerebrospinal fluid together with clonal B cell accumulation in the CSF and lesions of MS patients are among the main evidences of MS [115].
\nELISA was used for comparison of the relative levels of Abs against MBP in the sera of 12 healthy donors and 14 patients with SLE [77]. For healthy donors the concentrations of auto-Abs were not zero and varied from 0.02 to 0.16 A450 units; in average 0.09 ± 0.04 A450 units; similar value (0.09 ± 0.04 A450 units) was previously revealed for other 10 healthy volunteers [83]. Relative concentrations of anti-MBP Abs of SLE patients were changed from 0.27 to 0.54 A450 units, in average 0.38 ± 0.08 A450 units [77]. Using the same test system, it was previously revealed that the indexes of anti-MBP Abs for 25 MS patients are changed from 0.67 to 0.98 A450 units, in average 0.8 ± 0.1 A450 units [83, 84]. Thus, all SLE patients demonstrated in average ~4.2-fold higher level of anti-MBP Abs then healthy donors, but by a factor of ~2.1 lower level than MS patients.
\nElectrophoretically and immunologically homogeneous polyclonal IgGs were purified from the sera of SLE patients by sequential chromatography of serum proteins on Protein-G Sepharose using conditions removing nonspecifically bound proteins, followed by FPLC gel filtration in condition destroying immune complexes [77, 78] similarly to obtaining of MS IgGs [84–87]. It was shown that 150 kDa SLE IgGs are electrophoretically homogeneous and in contrast to Abs from healthy donors are active in the hydrolysis of MBP [55, 77]. To prove that MBP-hydrolyzing activity of SLE IgGs is their intrinsic property, we have checked the fulfilment of several known strict criteria described above including analysis of Ab protease activity after SDS-PAGE (Figure 7) [77]. In addition, it was shown that in contrast to canonical proteases, the SLE polyclonal IgGs separated using MBP-Sepharose specifically hydrolyzed only MBP (Figure 14A) but not many other tested control proteins (Figure 14B) [77].
\nThe hydrolysis of MBP (0.7 mg/ml) by 3 µg/ml IgGmix (mixture of IgGs of 12 SLE patients) separated by chromatography on MBP-Sepharose after incubation for 1 (lane 2), 2 (lane 3), 3 (lane 4), 14 (lane 5), 16 (lane 6), 18 (lane 7), and 24 h (lane 8) (A) [
Protease IgGs from the sera of ~95–100% of patients with different autoimmune pathologies [9, 66, 116], human milk Abs hydrolyzing casein [96, 117], Abs from AIDS patients hydrolyzing HSA, casein, and HIV reverse transcriptase [95] are serine-like proteases, whose activity is strongly decreased after their preincubation with serine protease-specific inhibitors PMSF. In addition, a high metal-dependent MBP-hydrolyzing activity for MS IgGs [86] and casein-hydrolyzing of human milk sIgAs [96] were recently revealed. It was shown that antiintegrase IgGs and IgMs of HIV-infected patients can contain abzymes hydrolyzing viral integrase of four types, resembling serine, thiol, metal-dependent, and acidic proteases, the ratio of which may be individual for every AIDS patient [97, 98].
\nIt was shown that preincubation of individual polyclonal SLE IgGs with specific inhibitor of thiol proteases iodoacetamide and of acidic proteases pepstatin A [77, 78] similarly to MS IgAs, IgGs, and IgMs [84–87] leads to a small effect (5–15%) on MBP hydrolysis by SLE IgGs. PMSF specifically inhibiting serine proteases and inhibitor of metalloproteases EDTA remarkably or significantly suppressed proteolytic activity of SLE IgGs (Figure 15A).
\nThe relative proteolytic activities of 12 different individual SLE IgGs in the hydrolysis of MBP (0.5 mg/ml) before (gray columns), after their preincubation with PMSF (white columns) or 35 mM EDTA (black columns) (A) [
The inhibition of Abz activity by EDTA is significantly greater than by PMSF. Overall, all individual SLE abzymes possess specific ratio of RAs in the presence of PMSF and EDTA (Figure 15A). Interestingly, polyclonal SLE abzymes was more sensitive to EDTA than MS Abs [77]. Catalytic heterogeneity of polyclonal abzymes with several different activities from patients with various autoimmune diseases and animals was shown in many papers [65, 80–82, 92]. The above data also demonstrate extreme heterogeneity of SLE abzymes hydrolyzing MBP. In addition, polyclonal MBP-hydrolyzing abzymes of every patient are characterized with specific dependence of RAs upon pH; the pH profile of each IgG is unique (Figure 15B and D). The effect of several different metal ions on the MBP-hydrolyzing activities of dialyzed against EDTA individual 12 SLE polyclonal IgGs was analyzed (Figure 16A and B; B is a continuation of A).
\nThe RAs of 12 SLE IgGs dialyzed against EDTA in the hydrolysis of MBP [
All 12 IgGs demonstrated the individual ratios of RAs in the presence of eight various metal ions. To analyze the “average” effect of different metal ions on SLE and MS IgGs, we have used SLE IgGmix and MS IgGmix before (Figure 16C) and after (Figure 16D) their dialysis against EDTA. Ca2+ was shown to the best activator of SLE IgGmix the effect of different metals decrease in the following order: Ca2+ > Co2+ ≥ Ni2+≥ Mg2+ ≥ Mn2+ ≥ Cu2+. Fe2+ did not activate SLE IgGmix, while Zn2+ inhibits its activity. MS IgGmix demonstrated a different order of the metal-dependent activity: Mg2+ > Mn2+ ≥ Cu2+ ≥ Ni2+ ≥ Co2+ ≥ Ca2+, while Fe2+ and Zn2+ slightly inhibit MBP hydrolysis (Figure 16C and D).
\nIn addition, the mixture of electrophoretically homogeneous IgGmix was separated to fractions of IgG1–IgG4 subclasses and to fractions of IgGs containing lambda- and kappa-type of light [76]. The immunological purity of IgGs of all types was revealed by ELISA; the preparations of IgG1, IgG2, IgG3, and IgG4 did not contain IgGs of other subclasses. The lambda - and kappa-IgGs and IgG1–IgG4 were active in the hydrolysis of MBP and their RAs and
IgG | \nContent, % | \nRAs (mg MBP/1 h) / mg of IgGs** | \nApparent | \nContribution to the total activity, %# |
---|---|---|---|---|
IgG, nonfractionated | \n100 | \n1.95 ± 0.05 | \n2.6 ± 0.07 | \n100 |
IgGs containing lambda- and kappa-types of light chains | ||||
kappa-IgG | \n44.6 ± 4.0 | \n2.1 ± 0.16* | \n2.8 ± 0.21 | \n48.4 ± 4.0 |
lambda-IgG | \n55.4 ± 5.0 | \n1.8 ± 0.15 | \n2.4 ± 0.20 | \n55.5 ± 4.3 |
IgGs of different subclasses | ||||
IgG1 | \n70.8 ± 2.0 | \n2.3 ± 0.11 | \n3.0 ± 0.14 | \n73.0 ± 3.4 |
IgG2 | \n20.6 ± 3.0 | \n2.0 ± 0.2 | \n2.7 ± 0.25 | \n19.1 ± 1.8 |
IgG3 | \n6.7 ± 1.5 | \n2.2 ± 0.08 | \n2.9 ± 0.10 | \n6.7 ± 0.3 |
IgG4 | \n1.9 ± 1.0 | \n1.3 ± 0.07 | \n1.7 ± 0.09 | \n1.2 ± 0.2 |
Relative specific MBP-hydrolyzing activities (RAs) of IgGs of different types and their relative contributions to the total activity of polyclonal IgGmix [76].
For each fraction, a mean of two repeats is used.
Relative activities at fixed 0.75 mg/ml concentration of MBP were estimated.
Apparent
Contribution of different IgGs to the total activity of nonfractionated Abs was calculated taking into account the relative content of these IgGs within polyclonal IgGmix and their RAs in the hydrolysis of MBP.
The pH dependence of RAs of MBP-hydrolyzing SLE kappa-and lambda-IgGs (A), as well IgG1, IgG2, IgG3, and IgG4 (B) [
These results clearly demonstrate that IgGs of all four subclasses are very heterogeneous and can consist of different sets of catalytic subfractions of polyclonal IgG having quite distinct pH dependencies. Figure 17 shows the relative influence of PMSF and EDTA on the MBP-hydrolyzing activity of different IgGs. The nonfractionated IgGs and lambda-IgGs demonstrated lower inhibition by PMSF than that for EDTA (Figure 17). The inhibition of serine-like and metal-dependent activities of kappa-IgGs were comparable. PMSF suppressed MBP-hydrolyzing activity of IgG3, IgG2, and IgG1 by 13–17%, while the decrease of this activity by EDTA was significantly greater, 30–45%. There was no noticeable PMSF effect on the IgG4 activity, while EDTA decreased its activity by ~65% (Figure 17). Thus, IgG1–IgG4, kappa-IgGs, and lambda-IgGs are characterized by specific ratios of metal-dependent and serine-like proteolytic activities.
\nThe cleavage site specificity of different IgG preparations in the case of four oligopeptides corresponding to four antigenic determinants of MBP was analyzed [76]. Overall, kappa-IgGs and lambda-IgGs, as well as IgG1–IgG4 demonstrated either different patterns of four oligopeptides cleavage, or at least stimulate the accumulation of the same products of the hydrolysis with different efficiency.
\nThe dialysis of IgGs caused a more pronounced decrease in the activity of kappa-IgGs than of lambda-IgGs [76]. Addition to the reaction mixtures of Ca2+ + Mg2+ or Ca2+ + Co2+ led to approximately comparable increase in the RAs of dialyzed lambda-IgG (1.6- to 1.7-fold), kappa-IgG (2.0- to 2.3-fold), and nonfractionated IgGs (1.7- to 1.8-fold). Ca2++Co2+ together cannot activate IgG1, while in the presence of Ca2+ + Mg2+ its activity increased by a factor of 1.6. Ca2+ + Co2+ increased the activity of IgG2 (~2.9-fold), IgG3 (~6.4-fold), and IgG4 (~6.0-fold). A significant increases in the RAs were revealed for Ca2+ + Mg2+ in the case of IgG3 (~3.5-fold), IgG4 (~4.4-fold), and IgG2 (~5.7-fold). While the Ca2+ + Mg2+ combination was the best for the activation of IgG2 and IgG1, IgG4, and IgG3 showed the highest activity in the presence of Ca2+ + Co2+. The ratios of RAs of all IgG preparations before and after their dialysis against EDTA, as well as in the presence of different metal ions, were individual for every preparation analyzed. These data indicate for an extreme Me2+-dependence diversity of different subclasses SLE IgGs hydrolyzing MBP.
\nThe extraordinary diversity of polyclonal abzymes with DNase, RNase, and proteolytic activities was shown not only in the case of SLE, but also other diseases [13–22]. Very unexpected enzyme properties have been discovered in the case of monoclonal abzymes of patients with SLE.
\nFor analysis of MBP-hydrolyzing activity of Abs, we have used the same phagemid library of kappa light chains [118–120] as for analysis of MLChs with DNase activity [111, 112]. The phage particles containing MLChs with different for MBP were separated by affinity chromatography on MBP-Sepharose (Figure 18A).
\nAffinity chromatography on MBP-Sepharose of phage particles: (− −) and (—) absorbance at 280 nm of particles corresponding plasmid respectively without and with kappa light chains cDNA (A) [
The pool of phage particles was distributed between 10 peaks eluted from the sorbent and all MLChs of fractions of 10 new small pools efficiently hydrolyzed MBP and four oligopeptides (OPs) corresponding to four immunodominant MBP sequences containing cleavage sites (Figure 18B). However, there were no any detectable particles peaks having considerable affinity for MBP after similar chromatography of phage particles with pCANTAB plasmid containing no cDNA of light chains (Figure 18A). Thus, the MLChs pools of all 10 phage particles fractions having different affinity to MBP contain both inactive and catalytically active light chains hydrolyzing MBP. Similar distribution all over the chromatography profiles was observed for polyclonal IgGs from SLE and MS patients in the case of their chromatography on MBP-Sepharose [76, 77, 86, 87].
\nPhage particles eluted from MBP-Sepharose with 0.5 M NaCl (peak 7, Figure 18A) were used for preparation of individual colonies. Overall, 72 of 440 individual colonies choosing in a random way were used for study of MBP-hydrolyzing activity. MLChs of 22 of 72 single colonies (~30%) possess MBP-hydrolyzing activity. All 22 recombinant catalytically active MLChs containing a sequence of 6 histidine residues interacting with Ni2+ ions and 5 MLChs without activity were purified by chromatography on charged with Ni2+ ions HiTrap chelating Sepharose and by following FPLC gel filtration. Then a mixture of equal amounts of 22 catalytically active monoclonal MLChs (act-MLChmix) and second mixture of five preparations without activity (inact-MLChmix) were prepared. The electrophoretical homogeneity of ~26- to 27-kDa inact-MLChmix and act-MLChmix was shown by SDS-PAGE with silver staining (Figure 19A, lane 1).
\nSDS-PAGE analysis of proteolytic activity (A) and homogeneity of act-MLChmix (7 µg) (B, lane 1) using a 5–16% gradient gel with following silver staining; the arrows indicate the positions of protein markers (B, lane 2) [
MLChmix was subjected to SDS-PAGE; its proteolytic activity was revealed after extraction of proteins from the separated gel slices only in the band corresponding to the MLCh (Figure 19A and B). Act-MLChmix demonstrated activity in the hydrolysis of MBP (Figure 19C, lane 4), while inact-MLChmix had no activity (Figure 19C, lane 2). Moreover, in contrast to canonical proteases cleaving all proteins, act-MLChmix hydrolyzes only MBP (Figure 19C, lane 4) but no other control proteins (Figure 19C, lanes 5–8). All 22 act-MLChs and 5 inact-MLChs showed positive answer with mouse Abs (conjugated with horseradish peroxidase) against light chains of human Abs at Western blotting and positive ELISA response using plates with immobilized MBP.
\nThe RAs in the hydrolysis of four different OPs were analyzed by TLC. Figure 19(D) and (E) demonstrates several typical examples of the OP19 and OP21 hydrolysis by different MLChs [118]. Initially, we have assumed that every of 22 MLChs corresponds to IgGs to one of four known specific MBP immunodominant sequences and that each MLCh can bind and hydrolyze only one of four OPs. At the same time, unexpected results were obtained. The RAs for 22 MLCh are summarized in Figure 19(F). All 22 MLChs hydrolyzed only three or four OPs and with significantly different efficiency in the case of every OP. Hydrolysis of OP17 MBP was very weak (~1–1.5%) except seven MLChs: 15 ≥ 10 ≥ 12 ≥ 1 ≥ 16 ≥ 20 ≥ 8 (1.6–7.1%) (Figure 19F). All MLChs except MLCh-22 hydrolyzed efficiently OP21 and several other OPs, while six other MLChs (8, 9, 10, 12, 13, and 14) demonstrated high activity only in the cleavage of OP21. Several MLChs (1–7, and 11) efficiently hydrolyzed OP19 and OP21, while MLCh-18 and 20 cleaved OP21 and OP25. Four recombinant MLChs (15, 17, 19, and 21) cleaved three OPs with relatively high efficiency, while MLCh-16 hydrolyzed all four OPs (Figure 19E). The ratios of the RAs in the hydrolysis of four OPs were specific for every MLCh (Figure 19E). OP21 and OP19 were shown to be the best substrates for most MLChs, while 15–22 MLChs better hydrolyzed OP25.
\nIn contrast to MS IgGs [76, 77, 84–87], SLE polyclonal abzymes with MBP-hydrolyzing activity are less sensitive to PMSF than to EDTA. The effect of PMSF and EDTA on the RAs of 22 different MLChs was analyzed [118]. Figure 20(A) shows that the 12 MLChs (1, 2, 3, 5, 7, 8, 12, 13, 15, 16, 17, and 19) are metal-dependent proteases; they cannot not remarkably decrease their proteolytic activity after incubation with PMSF, while EDTA significantly suppresses their MBP-hydrolyzing activity.
\nThe RAs of 22 MLChs in hydrolysis of MBP after Abzs preincubation with specific inhibitors of proteases of different type. MLChs (0.1 mg/ml) were preincubated without of other components (black bars), with 50 mM EDTA (gray bars) or with 1 mM PMSF (white bars); then aliquots of these mixtures were added to standard reaction mixtures (A and B) [
Four MLChs (4, 6, 9, and 11) demonstrate serine-like proteolytic activity; PMSF suppressed their activity, but there was no noticeable effect of EDTA (Figure 20B). PMSF suppressed protease activity of three MLChs (20, 21, and 22) by ~40%, and their inhibition by EDTA was to some extent comparable, 40–60% (Figure 20B). Thus, three MLChs (20–22) are characterized to some extent comparable ratios of metal-dependent and serine-like protease activities. A very intriguing situation was observed for three MLChs (18, 14, and 10); EDTA and PMSF do not remarkably decreased their proteolytic activity (Figure 20B). No significant suppression (5–15%) of MS and SLE polyclonal MBP-hydrolyzing abzymes by specific inhibitors of thiol proteases was revealed previously [76, 77, 84–87]. However, iodoacetamide inhibited integrase hydrolyzing activities of all polyclonal IgG and IgM preparations from HIV/AIDS patients by 12–99% [97, 98]. Proteolytic activities of three MLChs (18, 14, and 10) not inhibited by EDTA and PMSF were significantly suppressed by iodoacetamide, while there was no effect on the most of MLChs with metal-dependent and serine-like activities (for example, Figure 20C). Thus, these three MLChs (18, 14, and 10) are thiol proteases. Interestingly, but iodoacetamide significantly suppressed the activities of MLChs 17 and 12 (Figure 20C), which were also significantly inhibited by EDTA (Figure 20A). One can suppose that MLChs 17 and 12 may be MLChs, the active sites of which contain amino acid residues corresponding to metal-dependent and thiol proteases. A very surprising data were obtained for MLCh-22; its activity was significantly suppressed not only by EDTA and PMSF (Figure 20B), by also iodoacetamide (Figure 20C). The relative number of MLChs, which activity depend on iodoacetamide is only approximately 27% of all 22 MLChs, while at the same time, several of them possess metal-dependent and serine-like activities. Therefore, the relative contribution of thiol-like protease activity to a total MBP-hydrolyzing activity of polyclonal SLE and MS abzymes may be significantly lower than of Abzs with metal-dependent and serine-like proteolytic activities and, therefore, depending on the patient a relative contribution of thiol-like protease to the total activity may be about 5–15%, as found previously for polyclonal Abzs [76, 77, 84–87]. The effects of various metal ions on the protease activities of 22 MLChs were compared (Figure 21A and B; B is a continuation of A).
\nEffect of various metal ions on the RAs of 22 MLChs in the hydrolysis of MBP (A and B) [
Seven different metal ions did not effect on the activity of MLChs with serine-like (9, 6, and 4) and thiol-dependent (18, 14, and 10) activities. Five MLChs (19, 17, 13, 8, and 2) were only slightly activated by several Me2+ ions, while Ca2+ was the best activator. Two MLCh preparations (5 and 3) were Co2+ dependent, but preparation 15 was better stimulated by Ni2+, MLCh-16 and MLCh-20 were respectively Mn2+- and Zn2+-dependent (Figure 21). MLChs 22 and 12 were activated by two different metal ions, Zn2+ and Ca2+. Two MLChs were activated by three different Me2 ions: MLCh-7 (Ca2+ > Zn2+ > Co2+) and MLCh-1 (Ca2+ > Ni2+ > Mg2+). In addition, MLCh-21 was activated by four (Cu2+ > Ca2+ > Co2+ > Zn2+) metal ions. These data show the extreme Me2+-dependence diversity of IgGs from SLE patients and their light chains in the hydrolysis of MBP [118].
\nAll 22 MLCh preparations hydrolyzed efficiently MBP within a wide range of pHs (5.0–10), but in contrast to polyclonal SLE IgGs, they show mainly only one pH optimum [118]. Only the pH profile for preparation 4 demonstrates optimal pH at 5.7–5.9 and pronounced shoulder at pHs 6.5–7.5 (Figure 21C). The apparent
MLCh number | \nOptimal pHa | \nOptimal Me2+ cofactor | \nApparent |
---|---|---|---|
1 | \n8.0–8.2 | \n0.14 ± 0.01b | |
2 | \n7.6–7.8 | \n0.12 ± 0.01 | |
3 | \n7.6–7.8 | \n0.09 ± 0.007 | |
4 | \n5.7–5.9 | \nMe2+-independent | \n0.12 ± 0.008 |
5 | \n7.5–7.7 | \nCo2+ | \n0.24 ± 0.02 |
6 | \n7.0–7.2 | \nMe2+-independent | \n0.01 ± 0.001 |
7 | \n7.2–7.4 | \n0.07 ± 0.004 | |
8 | \n7.4–7.6 | \n0.17 ± 0.003 | |
9 | \n7.2–7.4 | \nMe2+-independent | \n0.03 ± 0.001 |
10 | \n8.1–8.3 | \nMe2+-independent | \n0.05 ± 0.002 |
11 | \n7.7–7.8 | \nMe2+-independent | \n0.09 ± 0.006 |
12 | \n7.8–8.0 | \n0.19 ± 0.001 | |
13 | \n6.1–6.3 | \n0.16 ± 0.001 | |
14 | \n7.0–7.2 | \nMe2+-independent | \n0.06 ± 0.0003 |
15 | \n7.0–7.2 | \n0.17 ± 0.007 | |
16 | \n6.9–7.1 | \n0.12 ± 0.001 | |
17 | \n8.2–8.4 | \n0.17 ± 0.001 | |
18 | \n6.2–6.4 | \nMe2+-independent | \n0.05 ± 0.0007 |
19 | \n6.7–6.9 | \n0.1 ± 0.01 | |
20 | \n8.2–8.4 | \n0.09 ± 0.006 | |
21 | \n6.9–7.1 | \n0.18 ± 0.007 | |
22 | \n7.8–8.0 | \n0.11 ± 0.006 |
The optimal pH values, optimal metal cofactors, and apparent
For each value, a mean ± S.E. of two/three measurements is reported.
Optimal pH of reaction mixtures and optimal metal cofactor (given in bold) were used for every of MLCh preparations; the apparent
The best metal activator is given in bold, while alternative cofactors demonstrating relatively high activation are given in parenthesis.
On the next step we analyzed in more detail three additional MLChs (numbers 23–25) corresponding to peak 7 eluted from MBP-Sepharose with 0.5 M NaCl (Figure 18A) [119, 120]. These three MLChs were purified and characterized in detail exactly similar to above described 22 preparations [118]. The DNA sequence of NGTA1-Me-pro demonstrated high identity to germline VL genes of IgLV8-61*02, IgLV8-61*01, and IgLV8-61*03IGKV1 (90% of identity) [120]. DNA sequence of NGTA2-Me-pro-Tr indicated high identity with germline VL gene IGKJ1*01 (100%), IGKJ4*01 (95.7%), IGKJ4*02 (91.2%), IGKV1-5*03 (87.9%), IGKV1-5*01 (86.2% of identity), and IGKV1-5*02 (85.6%) [119]. DNA sequence of NGTA3-pro-DNase has similarity with germline DNA sequence of light chains of several IgGs: IGKJ1*01 (100% of identity), IGKJ4*01 (95.7%), IGKJ4*02 (91.2%); IGKV1-5*03 (79.8% of identity), IGKV1-5*02 (78.4%), and IGKV1-5*01 (78.4%) [Timofeeva, Nevinsky, personal communication]. Thus, all three MLChs were shown to be typical light chain of Abs [119, 120, personal communication].
\nNGTA1-Me-pro was shown to be a specific metalloprotease; only EDTA efficiently inhibits its activity, while specific inhibitors of thiol-, serine-, and acidic-like proteases did not suppress its MBP-hydrolyzing activity (Figure 22A) [120].
\nThe RAs of NGTA1-Me-pro in the hydrolysis of MBP before and after its preincubation with specific inhibitors of various type proteases [
Seven various metal ions increase NGTA1-Me-pro activity in the following order: Ca2+ > Mg2+ > Ni2+ ≥ Zn2+ ≥ Co2+ ≥ Mn2+ > Cu2+ (Figure 22B). NGTA1-Me-pro demonstrated two different very well expressed pH optima at pH 6.0 and 8.5 (Figure 22C). Figure 22(D) indicates that at pH 6.0 MLCh has optimum at ~6 mM, when at pH 8.5 at 1 mM CaCl2. The apparent values of
MLCh NGTA2-Me-pro-Tr demonstrated two different activities: trypsin-like and metalloprotease. Figure 23(A) shows that NGTA2-Me-pro-Tr is not sensitive to pepstatin and iodoacetamide [119]. Preincubation of this MLCh with specific inhibitor of serine-like proteases results in a decrease of its activity for 42 ± 4%.
\nThe RAs of the activity of NGTA2-Me-pro-Tr in the hydrolysis of MBP after its preincubation with specific inhibitors of different type proteases (A) [
Intact polyclonal Abs interact with various metal ions and they do not lose completely intrinsically bound ions during the standard purification procedures [121]. Addition of EDTA to NGTA2-Me-pro-Tr containing only intrinsically bound Me2+-ions results in a decrease in its activity for 58 ± 5% (Figure 23A) [119]. Average serine-like activity of NGTA2-Me-pro-Tr containing only intrinsically bound Me2+ ions was ~1.4-fold lower than its Me2+-dependent protease activity. Seven various external metal ions activate this MLCh in the following order: Ca2+ ≥ Mn2+ ≥ Mg2+ > Co2+> Ni2+ ≥ Cu2+ ≥ Zn2+ (Figure 23B). After NGTA2-Me-pro-Tr treatment with PMSF, its metalloprotease activity demonstrated pH optimum at 6.5–6.6 (Figure 23C). After dialysis of this MLCh against EDTA or in the presence of EDTA, serine-like protease activity showed pH optimum at 7.4–7.5. Figure 23(D) demonstrates that the increase in PMSF concentration results in a complete suppression of the activity at pH 7.5 in the presence of 50 mM EDTA, conditions corresponding to serine-like activity. NGTA2-Me-pro-Tr containing no intrinsic metal ions demonstrated in the absence of external metal ions at pH 7.5
It should be emphasized that all recombinant MLChs were obtained using affinity chromatography of phage particles on MBP-Sepharose and all electrophoretically homogeneous preparations of MLChs have affinity for MBP-Sepharose; similar to phage particles homogeneous MLChs were eluded from the sorbet by 0.5 M NaCl. Taking this into account, a very unexpected result was obtained from the analysis of enzymatic activities of NGTA3-pro-DNase [Timofeeva and Nevinsky, personal communication].
\nThe homogeneity of ~26–27-kDa NGTA3-pro-DNase was confirmed using SDS-PAGE with following silver staining (Figure 24B, lane 1). NGTA3-pro-DNase demonstrated positive answer with horseradish peroxidase conjugated with mouse IgGs against human Abs light chains at Western blotting and positive ELISA answer using plates with immobilized MBP and DNA.
\nSDS-PAGE analysis of MBP- and DNA-hydrolyzing activities (A) and homogeneity of NGTA3-pro-DNase (7 μg) using a reducing 5–16% gradient gel followed by silver staining (B, lane 1); the arrows (B, lane 2) indicate the positions of protein markers. After SDS-PAGE the gel was incubated using conditions for renaturation of NGTA3-pro-DNase. The relative MBP- and DNA-hydrolyzing activity (%) was revealed using the extracts of 2- to 3-mm gel fragments (A). The activity of NGTA3-pro-DNase corresponding to a complete hydrolysis of 0.5 mg/ml MBP (or 18 nM scDNA) after 24 h of incubation of 25 μl reaction mixture containing 10 μl of the gel extracts was taken for 100%. SDS-PAGE analysis of hydrolysis of MBP by inact-MLChmix (lane 1) or NGTA3-pro-DNase (lanes 2 and 3, different time of incubation) (C). Hydrolysis of control proteins (0.5 mg/ml) by inact-MLChmix and NGTA3-pro-DNase was analyzed: human albumin (lanes 4 and 5) and lactoferrin from human milk (lanes 6 and 7) (C). The mixtures were incubated for 6 h with inact-MLChmix (lanes 4 and 6), or NGTA3-pro-DNase (lanes 5 and 7). All lanes C correspond to different proteins incubated alone without MLChs, while lane C1- to standard protein markers. DNase activity of NGTA3-pro-DNase and two control MLCh1 and MLCh2 (10 nM) was analyzed in the presence of 5 mM MgCl2 (D); lane C corresponds to scDNA incubated alone.
After SDS-PAGE, MBP-hydrolyzing activity was revealed only in the band corresponding to the light chains in the presence of CaCl2 (o) and in the absence of external metal ions (□); the positions of proteolytic (o, □) and DNase (x) activities of MLCh are coincided (Figure 24A). NGTA3-pro-DNase hydrolyzed specifically only MBP and not hydrolyzed foreign control proteins (Figure 24C).
\nOnly one (NGTA3-pro-DNase) of 25 recombinant MLChs analyzed by us efficiently hydrolyzed not only MBP, but also DNA (for example, Figure 24D). DNase activity of NGTA3-pro-DNase was determined
NGTA3-pro-DNase containing intrinsic metal ions was not sensitive to treatment with iodoacetamide and pepstatin, while its preincubation with PMSF led to decrease in the activity for 67 ± 5% (Figure 25A).
\nThe RAs of MBP-hydrolyzing activity of NGTA3-pro-DNase after its preincubation with specific inhibitors of different types proteases (A). MLCh (0.1 mg/ml) was preincubated alone (control), in the presence of iodoacetamide, PMSF, pepstatin, or EDTA, and then 1.5 μl added to 29 μl of standard reaction mixture (A). The RA of NGTA1-Me-pro before its preincubation with various inhibitors was taken as 100%. Effect of EDTA and different metal ions (2 mM) on the RA of MLCh is shown (B). Dependence of RA of MBP-hydrolyzing activity of NGTA1-Me-pro on pH of reaction mixture before and after its treatment with EDTA and PMSF is given (C).
The dialysis of NGTA3-pro-DNase containing only intrinsically bound Me2+ ions against EDTA or addition of EDTA to reaction mixture led to a decrease in its activity for 33 ± 3% (Figure 25A). And average Me2+-dependent protease activity of MLCh containing only intrinsically bound Me2+ ions was approximately 2.0-fold lower (Figure 25A), but after addition of external Ca2+ ions became to be 2.2-fold higher than its serine-like activity (Figure 25B). Seven various external metal ions activate NGTA3-pro-DNase in the following order: Ca2+ ≥ Ni2+ > Co2+ ~ Mn2+ ≥ Cu2+ ~ Zn2+ ≥ Mg2+ (Figure 25B). An optimal concentration of CaCl2, which is the best activator of this MLCh, was 3 mM. NGTA3-pro-DNase demonstrates two different optimal pHs (Figure 25C). After treatment of MLCh with PMSF, its metalloprotease activity was maximal at pH 8.6, while in the presence of EDTA, serine-like protease activity demonstrated pH optimum at 7.0 (Figure 25B). NGTA3-pro-DNase treated with PMSF in the presence of 3 mM CaCl2 (pH 7.0) demonstrated
It is known that Mg2+ (10 mM) is optimal cofactor of DNase I, while other Me2+ ions very weakly activate DNase I [109, 110]. Optimal concentration for Mn2+, Mg2+, and Ni2+ in activation NGTA3-pro-DNase was ~4–5 mM, for Ca2+ and Zn2+ 2 mM, while Co2+ and Cu2+ activate MLCh up to 10 mM concentration. DNase activity increased in the presence of metal ions in the following order: Mn2+ ≈ Co2+ ≥ Mg2+ > Cu2+ ≈ Ni2+ ≥ Ca2+ > Zn2+), which is completely different in comparing with that for DNases I and other recombinant MLChs analyzed.
\nDNase activity for NGTA3-pro-DNase in the presence of Mg2+ or Mn2+ at fixed concentration (5 mM) was increased at optimal concentrations of NaCl or KCl (30–40 mM) for only 27–28%. In optimal conditions, NGTA3-pro-DNase demonstrated well expressed optima at pH 6.5–6.6. The
In several articles, it was demonstrated that polyclonal RNA-, DNA-, MBP- integrase-, and oligosaccharide-hydrolyzing antibodies of different classes and subclasses from patients with SLE, MS, AIDS, and other diseases are very catalytically heterogeneous. These abzymes can contain lambda- and kappa- types of light chains, may be of different subclasses (IgG1–IgG4), can demonstrate different affinity for specific sorbents and free antigens-substrates, very different pH optima, and may be independent or dependent on metal ions. Different abzymes can catalyze the hydrolysis of MBP, HIV integrase, and other proteins as serine-, thiol-, and acidic-like or metalloproteases. Various IgGs of four subclasses (IgG1–IgG4) and/or IgAs and IgMs from the sera of patients with autoimmune and viral diseases are catalytically active in the hydrolysis of RNA, DNA, oligosaccharides, and various proteins with their different contribution to the total activity of the Abs in the hydrolysis of these substrates in the case of every individual patient.
\nAt the same time, the analysis of polyclonal antibodies does not allow to obtain detail characteristics of monoclonal abzymes entering to small pools of polyclonal antibodies separated by affinity chromatography on sorbents with different immobilized antigens-substrates. As it was shown on the example of polyclonal IgGs with DNase and MBP-hydrolyzing activities from sera of SLE and MS patients, elution of Abs by a NaCl concentration gradient leads to their distribution all over the chromatography profiles. In this case, each eluted Ab fraction contains abzymes with comparable affinity for immobilized ligand, but demonstrating a significant diversity of various enzymatic properties described above. These data are strong evidence of exceptional diversity abzymes in the blood of some patients with SLE, MS, and other diseases. In this regard, it should be mentioned that theoretically immune system of human can produce up to 106 different Abs against one antigen. It is evident that all theoretically possible variants of antibodies are in reality not realized and much less than one million. However, in the case of some patients, a possible number of abzymes can be very large. In our studies, we used a cDNA library only kappa light chains of Abs from three patients with SLE [111, 112, 118–120]. We have analyzed only 45 of 451 single of colonies corresponding one peak eluted from DNA-cellulose with 0.5 M NaCl and 33 of 687 colonies of peak eluded with acidic buffer. In the first case 15 of 45 (~33%) [111] and in the second 19 of 33 MLChs (58%) demonstrated DNase activity [112]. For analysis of MBP-hydrolyzing activity, we have used 72 of 440 individual colonies corresponding to phage particles eluted from MBP-Sepharose with 0.5 M NaCl; 25 of 72 MLChs (~35%) effectively hydrolyzed MBP [118–120]. Since we analyzed abzymes corresponding only one or two of ≥10 phage particles, it is obvious that the number of MLChs with DNase and MBP-hydrolyzing activity with very different enzymatic properties may be at least ≥ 500–1000.
\nThe question is why many abzyme with nuclease and protease activities exist in SLE and other AI patients. First, immunization of autoimmune mice leads to a dramatically higher incidence of Abzs with a higher activity comparing to conventionally used normal mouse strains [51, 52]. The immune response to RNA and DNA and their complexes with histones and other proteins only partially depends on the length and sequence of nucleic acid [123, 124]. In addition, antiidiotypic Abs against the active centres of different DNA- and RNA-dependent enzymes can also possess catalytic activity. We have shown that polyclonal nuclease abzymes of autoimmune patients are usually different cocktails of Abzs against DNA and RNA and their complexes with proteins as well as antiidiotypic Abzs to active centers of various DNA- and RNA-hydrolyzing enzymes [13–22].
\nIt is possible to explain to some extent in a similar way the exceptional diversity of abzymes hydrolyzing MBP and other proteins. At the same time, possible ways of production of monoclonal abzymes having two or even three different catalytic centers have a special interest. It should be noted that the known antigenic determinants of different proteins are usually relatively long and the active centers of some abzymes with two activities can correspond at once to variable parts of the antibodies to different contiguous parts of these determinants. One cannot exclude that metal-dependent active centers may be against specific part of protein antigenic determinants bound with one or several metal ions.
\nThe second question is why NGTA3-pro-DNase against MBP can hydrolyze DNA. It is believed that MBP and anti-MBP Abs cannot interact with DNA or RNA. However, it was recently shown that anti-MBP IgGs can efficiently interact with nucleic acids [125]. Using quenching of MBP tryptophan fluorescence emission, we have shown that MBP bind oligonucleotides showing two
As mentioned above, DNA-hydrolyzing Bence-Jones proteins [60] and DNase abzymes of patients with SLE [59] and MS [16] are dangerous since they are cytotoxic, can penetrate to cell nuclear, and hydrolyze nuclear DNA resulting in cell apoptosis. Abzymes against vasoactive peptide are harmful since they decrease in the concentration of the peptide and have an important negative role in pathogenesis of patients with asthma [126]. RAs of DNase abzymes of patients with Hashimoto thyroiditis well correlate with different immunological and biochemical indices of this disease including a concentration of thyroid hormones, while decrease in their activity is related to decrease in thyroid gland damage and improvement of the clinical status [105]. Protease IgGs of patients with sepsis participate in the control of disseminated microvascular thrombosis and play important role in recovery from the disease [127]. Thus, various abzymes can play both a negative and positive role in the pathogenesis of SLE and other autoimmune or viral diseases. Meanwhile, it should be mentioned that in the later stages of SLE, MS, and other diseases, the blood of these patients contains abzymes not only with DNase and MBP-hydrolyzing activities, but also hydrolyzing other proteins, oligosaccharides, and lipids [13–22].
\nAs it was shown in the example of Hashimoto thyroiditis production of harmful abzymes can be suppressed by using therapy with suppressing immune system drug plaquenil [102]. In MS and SLE, anti-MBP abzymes with proteolytic activity can attack MBP of the myelin-proteolipid sheath of axons. The established MS drug Copaxone was shown to be a specific inhibitor of abzymes with MBP-hydrolyzing activity [128]. One cannot exclude that the same drugs can be used for the treatment of SLE and other autoimmune diseases, which characterized by high level of abzymes with nuclease and MBP-hydrolyzing activities.
\nThe presence of anti-DNA Abs is known as the main important diagnostic index for SLE. High-affinity anti-DNA Abs was recently shown to be major component of the intrathecal IgG in cerebrospinal fluid and brain of MS patients [48]. Moreover, DNase abzymes from SLE and MS patients are cytotoxic and induce cell death by apoptosis [16, 59]. The sera of patients with SLE and MS patients contain different free light chains [61, 62]. Therefore, we propose that exceptional diverse of intact antibodies and their free light chains hydrolyzing DNA, MBP, nucleotides, and polysaccharides may cooperatively all together promote important neuropathologic pathogenic mechanisms in SLE and MS.
\nOur data on the study of abzymes production in SLE patients associated with the change in profile differentiation of brain stem cells seem to be very important for understanding possible mechanisms of various autoimmune diseases development.
\nThis research was made possible by grant from the Russian Science Foundation (no 16-15-10103 to G.A. Nevinsky).
\nAbs | Antibodies |
Abzs | Abzymes, or catalytically active antibodies |
AI | Autoimmune |
AD | Autoimmune disease |
AIDS | Acquired immunodeficiency syndrome |
a/u | Arbitrary units |
BSA | Bovine serum albumin |
HSCs | Hematopoietic stem cells |
CBA | (CBAxC57BL)F1 mice |
MBP | Human myelin basic protein |
OP-17, OP-19, OP-21, and OP-25 | 17–25mer oligopeptides corresponding to four known MBP cleavage sites |
MS | Multiple sclerosis |
m-BSA | Methylated BSA |
nat-DNA and den-DNA | Native and denatured DNA, respectively |
OP | Oligopeptide |
SLE | Systemic lupus erythematosus |
SDS-PAGE | SDS-polyacrylamide gel electrophoresis |
RA | Relative activity |
“Assessment is a central component of the teaching and learning” process. It is defined as “the systematic collection and analysis of information to improve student learning” [1]. Test (exam) is a part of student assessment and should be “An objective and standardized measure of a sample of behavior” [2]. Item analysis is a post-examination evaluation and can provide information about the quality of tests.
Item analysis is a statistical analysis of the student’s responses on a test. Collection and summarization of students’ responses can provide quantitative objective information that is useful in deciding the quality of the test items and increasing the assessment’s efficiency [3, 4]. Also, Item analysis “investigates the performance of items considered individually either in relation to some external criterion or the remaining items on the test” [5].
Any educational test should measure students’ achievement in content material. Also, it leads to an overall assessment of students’ development to decide their academic status [6, 7].
The importance of item analysis is determined by the objective of the assessment [8]. In summative assessment, the assessment results should be reliable and valid because incorrect decisions about the academic status will lead to negative consequences [9]. While for the formative evaluation where the target is students learning, the item analysis has no much importance in giving feedback about items construction to test composers.
In literature, many reasons were reported for the conduction of item analysis, including examining if the item is functioning as intended, did it assess the required concepts (content)?, did it discriminate between those who master the content material and those who were not? was it within the acceptable level of difficulty?, whether the distracters are functioning or not? [10, 11].
Many factors can affect item analysis and hence its interpretation [8]. Difficulty and discrimination indices were constantly changing per administration and influenced by the ability and number of the examinee, the number of items, and the quality of instructions [8, 12].
Whatever the exam or test blueprinting (item selection) method, exam items remain a sample of the needed content material. The number of items (item sampling) carries excellent importance because one cannot ask about all contents. With a too-small number of items, the results may not be enough to reflect true student ability [8]. Technical item flaws are divided into two major types, test wiseness, and irrelevant difficulty. Test wiseness flaws can result in more easy items. Faults related to irrelevant difficulty can result in more challenging items unrelated to the content under assessment. It was reported that item analysis of exam with 200 examinees is stable, and with fewer than 100 examinees should be interpreted with caution (item difficulty or item discrimination index). While Downing and Yudkowsky described that even for a small number of the examinee (e.g., 30) still, the item analysis can provide a piece of a helpful information to improve item [13, 14].
The item or psychometric analysis parameters include difficulty index, reliability, discrimination index, distractor efficiency [2]. The descriptive statistics of the exam are important and can provide helpful generalized information [2]. The descriptive statistics include scores frequency, the mean, the mode, the median, and the standard deviation.
Cronbach’s alpha (KR20) is widely accepted and used estimate of test reliability (the internal consistency) and reported to be superior to the split-half estimate [15, 16]. Although validity and reliability are closely associated, the reliability of an assessment does not depend on its validity [16, 17]. Coefficient alpha is known to be equal to Kr-20 if the item has a single answer, such as in the case of type A MCQs or binary [18, 19, 20, 21].
Coefficient alpha reflects the degree to which item response scores correlate with total test scores [15]. It also describes the degree to which items in the exam measure the same concept or construct [22]. Therefore, it is connected to the inter-relatedness and dimensionality of the items within the exam [16, 20]. Cronbach’s alpha is affected by exam time, the number and inter-relation of the items (dimensionality) and easy or hard, poorly written or confusing items, Variations in examinee responses, curriculum content not reflected in the test, Testing conditions, and Errors in recording or scoring [22, 23, 24]. The value of alpha is decreased in the exam with fewer items and increased if items assessing the same concept (unidimensionality of the exam) [16]. Other factors were reported to impact alpha value, such as item difficulty, number of the examinee, and student performance in the exam time. It was argued that very high alpha values could indicate lengthy exams, parallel items, or a narrow coverage of the content material [22].
The alpha value of the exam can be increased by increasing the number of items with a high p-value (difficulty index). It was reported that items with moderate difficulty could maximize alpha value and while those with zero difficulties or 100 can minimize it [15]. In the same way, deletion of faulty items can increase alpha value. It should be considered that repetition of items in the same exam or using items assessing the same concept can increase alpha value.
The interpretation of reliability is the correlation of the test with itself. When the estimate of reliability increases, the portion of a test score related to the error will decrease. Wise interpretation of alpha needs an understanding of the interrelatedness of items and whether the items measure a single latent trait or construct. Exam or test with different content materials such as integrated courses, for example, in the musculoskeletal system course, although is dominated by anatomy it contains other subjects of basic medical and clinical sciences that have different contains. Therefore, interpretation of such a course exam needs deep looks beyond the alpha figure. It was reported that KR20 of 0.7 is acceptable to short test (less than 50 items) and KR20 of 0.8 for an extended test (more than 50 item-test) [25]. Moreover, it was documented that a multidimensional exam does not have a lower (Table 1) alpha value than a unidimensional one [26].
Author | Interpretation of Cronbach’s alpha (KR20) |
---|---|
Robinson, Shaver et al. [29] | ≥0.80 Exemplary |
0.70–0.79 Extensive | |
0.60–0.69 Moderate | |
<0.60 Minimal | |
Cicchetti [30] | <0.70 Unacceptable |
0.70–0.80 Fair | |
0.80–0.90 Good | |
< 0 .90 Excellent | |
Axelson and Kreiter [31] | >0.90 is needed for very high stakes tests (e.g., licensure, certification exams) |
0.80–0.89 is acceptable for moderate stakes tests (e.g., end-of-year summative exams in medical school, end-of-course exams) | |
0.70–0.79 would be acceptable for lower stakes assessments (e.g., formative or summative classroom-type assessments created and administered by local faculty | |
<0.70 might be useful as one component of an overall composite score. | |
Obon and Rey [12] | >0.90 Excellent reliability |
0.80–0.90 Very good for a classroom test | |
0.70–0.80 good for a classroom test | |
0.60–0.70 Somewhat low (The test needs to be supplemented by other measure) | |
0.50–0.60 Suggests need for revision of test (unless it is quite short, ten or fewer Items). | |
0.50 < Questionable reliability. | |
Hassan and Hod [32] | > 0.7 is excellent |
0.6–0.7 is acceptable | |
−0.5-0.6 is poor | |
< 0.5 is unacceptable | |
< 0.30 is unreliable |
Reference values and interpretation of Cronbach’s alpha (KR20).
A low alpha value can be due to a smaller number of items, reduced interrelatedness between items, or heterogeneous constructs [22]. A high value of alpha can suggest exam reliability, and some items are non-functional as they are testing the same content but in a different guise or repeated ones [16, 22]. Also, a high value indicates items with high interrelatedness, indicating a limited coverage of the content materials [22].
Adding new items with an acceptable difficulty index, high discrimination power and distractor efficiency can increase the test reliability [22, 27, 28]. In addition, deletion of faulty items or those with low or very high p-value can improve Cronbach’s alpha. Items with poor correlation or are not related should be revised or discarded from the exam.
Commonly are formed of a stem with or without leading question and five or four alternatives (type A MCQs). Among item’s alternatives, only one is the key answer and others are called distractors [4]. Distractors should carry or convey a miss concept about the key answer and appear plausible. The distractors should appear similar to the key answer in terms of the used words, grammatical form, style, and length [19]. Distractor efficiency (DE) is the ability of incorrect answers to distract the students [12].
A functional distractor (FD) the distractor that is selected by 5% or more of the examinee [4, 33]. At the same time, those chosen by less than 5% of the examinee are considered non-functional (NFD) [4]. In comparison, other authors reported 1% of the examinee as the demarcation of functional distractors [34, 35]. Commonly items are categorized based on the numbers of NFDs in the item (Table 2) [12, 29, 36, 37].
Number of NFD | Percentage | Interpritation |
---|---|---|
3 | 0 | Poor |
2 | 33.3 | Moderate |
1 | 66.6 | Good |
0 | 100 | Excellent |
Classification of items according to thee number of the nonfunctional distractors (NFD).
The occurrence of NFD makes the item easier and reduces its discrimination power, while FD distractors are making it more difficult [36, 38]. It was reported that non-functional distractors are negatively correlating with reliability [38]. The presence of non-functional distractors can be related to two main causes. First is the training and construction ability of the item writer or composer. Second, the miss-match between the target content and the possible number of a distractor created. Thus, training and more effort in item writing and construction can decrease NFDs [36]. Other causes were related to NFDs, including the low cognitive level of the item, irrelevant or limited number of plausible distractors, or presence of logic cues [39]. Another possibility of NFDs is mastering the content material of the item, and students can identify the distractor as the wrong one. If no other cause (s) for NFDs, they should be removed or changed with a more plausible option because it has no contribution to the measurement of the test [12]. If a distractor is selected more frequently than the key answer by a higher-scoring examinee, this may indicate poor constructions or a misleading question or miss or double-keyed [12, 40]. In this, concerning the use of three options is more practical than four, does not affect reliability, and does not affect the discrimination index significantly [29, 35, 36, 37].
Furthermore, it was reported that there is no psychometric reason that all items in the exam should have the same number of distractors [29, 41]. The required number of options in an item should be considered according to the content material from which plausible distractors can be developed [33, 40, 42].
Reducing the number of options/distractors will result in other important benefits such as reducing the answering time of the test and safe time can be used to cover more content material, reduce the burden on item composers, and have items with more acceptable parameters [43, 44].
Puthiaparampil et al. reported a non-high significant negative and positive correlation between the number of functional distractors and difficulty and discrimination indices, respectively [34]. While a significant positive correlation was reported between the DIF and the number of NFDs [45].
Many authors concluded that no predictable relationship between DE and difficulty index and discrimination index [27, 29, 40, 46, 47]. In addition Licona-Chávez et al. did not find a parallel performance between DE and other parameters of item analysis including Cronbach alpha [46]. In contrast, some authors claimed that low DE decreases the difficulty index [47, 48].
Restoring the optimal DE of the item can be achieved by identifying flaws related to the NFDs and correcting them or removing the NFDs from the item [39].
The item difficulty (easiness, facility index, P-value) is the percentage of students who answered an item correctly [6, 40]. The difficulty index ranges from 0 to 100, whereas the higher the values indicate, the easier the question and the low value represents the difficulty of hard items. The ideal (optimal) difficulty levels for type A MCQs is varying according to the number of the options (Table 3) [49, 50]. The range of items difficulty can be categorized into difficult, moderate, and easy. Easy and difficult items were reported to have very little discrimination power [48]. Item difficulty is related to the item and the examinee that took the test in the given time [24]. Thus, reusing of the item depending on its difficulty index should be controlled. Some authors found that difficulty indices of items assessing high cognitive levels in Bloom’s taxonomy such as evaluation, explanation, analysis, and synthesis are lower than those assessing remembering, understanding, and applying [51, 52].
The ideal (optimal) difficulty level (for tests with 100 items).
Author | Difficulty index | Interpretation |
---|---|---|
Uddin et al. [50] | >80% | Easy |
30–80% | Moderate | |
<30% | Difficult | |
Kaur, Singla et al. [56] | >80 | Easy |
40–80 | Moderate | |
<39 | Difficult | |
Sugianto [6] | 90 | Easy |
50 | Moderate | |
10 | Difficult | |
Date, Borkar et al. [37] and Kumar, Jaipurkar et al. [36] | <30 | Too difficult |
>70% | Too easy | |
50–60% | Excellent/ideal | |
30–70% | Good/acceptable/average | |
Obon and Rey [12] | > 0.76 | Easy (Revise or Discard) |
0.26–0.75 | Right difficult (Retain) | |
0–0.25 | Difficult (Revise or Discard) | |
Bhat and Prasad [57] | >70% | Easy |
30–70% | Good | |
<30% | Difficult |
Reference values and interpretation of difficulty index (p-value).
During item or exam construction, the constructor should aim for acceptably level of difficulty [6]. Sugianto reported that items within the exam could be distributed according to difficulty to moderate level (40%), easy and challenging levels (20%), and easier and more challenging levels (10%) [6]. Other authors reported that most items should be of moderate difficulty or 5% should be in the difficult range [50, 53]. Some authors found that difficulty indices of items assessing high cognitive levels in Bloom’s taxonomy such as evaluation, explanation, analysis, and synthesis are lower than those assessing remembering, understanding, and applying [51, 52]. Regarding the general arrangement of test or examination, easy items start first then are followed by difficult ones. At the same time, in the case of diagnostic assessment, the sequence of the learning material is more important [6, 7].
Easy and difficult items affect the item’s ability to discriminate between students and show low discrimination power. Some reports described a negative correlation between exam reliability and difficult and easy items [38]. Oermann et al. reported that educationalists must be careful in deleting items with poor DIF because the number of items has more effect on test validity [54]. It is recommended that difficult items should be reviewed for the possible technical and content causes [50]. Possible causes of low difficulty index include uncovered (taught) content material, challenging items, missed key or no correct answer among the item options [55]. Easy items (high
In literature including medical education, many ranges of difficulty indices were reported (Table 4).
Item discrimination (DI) is the ability of an item to discriminate between higher achiever (good) students and low ones. It was defined as “stated that item discrimination is a statistic that indicates the degree to which an item separates the students who performed well from those who did poorly on the test as a whole” [6]. The discrimination power of an item is calculated by categorizing the examinee into upper 27% and lower 27% according to their total test score. The difference between the upper and lower group is divided by the number of the examinee in the upper group or the larger group or by half of the total number of the examinee or even by the total number [4, 6, 58, 59]. Obon and Rey [12] calculated the discrimination index as the difference of difficulty index between the upper and lower groups [12]. In literature, both 25 and 27% were reported as possible percentages of examinee categorization [60, 61]. The 27% is commonly used to maximize differences in normal distributions and increase the number of examinees in each category. The discrimination index range from 1.0 to −1.0. The positive discrimination index indicates that high achievers answer the item correctly more than those in the lower ones, which is desirable. The negative discrimination index reflects that lower achiever examinees answer the item more correctly, while zero discrimination indicates equal numbers of students in the upper and lower groups [36, 37]. Negative discrimination is thought to be due to item flaws or inefficient distractors, miss keys, ambiguous wording, gray areas of opinion, and areas of controversy [12, 62]. Nevid and McClelland [52] reported that items assessing evaluation and explanation domains could discriminate between high and low performers, while Kim et al. [51] comments that items assessing remembering and understanding levels have low discrimination power [52, 54].
It was reported that discrimination indices are positively associated with difficulty index and distractor efficiency [39, 63]. The discrimination power of the item is reduced by the increased number of non-functional distractors [36].
A test with poor discriminating power will not provide a reliable interpretation of the examinee’s actual ability [6, 64]. In addition, discrimination power will not indicate item validity, and deletion of items with poor discrimination power negatively impacts validity due to a decrease in the item number [65].
Discrimination coefficients can evaluate item discrimination. The discrimination coefficients include point biserial correlation, biserial correlation, and phi coefficient. Although point biserial correlation is used interchangeably with the discrimination index, discrimination coefficients are considered superior to the discrimination index [24]. The superiority came from the fact that discrimination coefficients are calculated using all examinees’ responses in the item rather than only 54% of the examinees such as in the discrimination index.
The difference between Point-biserial correlation (rBP) and discrimination indexes is that rBP is the correlation between an item in the exam and the overall student score [2, 66]. In cases of highly discriminating items, the examinees who responded to the item correctly also did well on the test. In general, the examinees who responded to the item incorrectly also tended to perform poorly on the overall test. It was suggested that point biserial can express the predictive validity better than Biserial correlation coefficients [61, 67].
Discrimination power of items more than 0.15 was reported as evidence of item validity [50, 53]. While any item with less than 0.15 or negative should be reviewed [50] (Table 5).
Author | Discrimination power | Interpretation |
---|---|---|
Elfaki, Bahamdan et al. [53] | ≥0.35 | Excellent |
0.25–0.34 | Good | |
0.21–0.24 | Acceptable | |
≤ 0.20 | Poor | |
Obon and Rey [12] | ≥ 0.50 | Very Good Item (Definitely Retain) |
0.40–0.49 | Good Item (Very Usable) | |
0.30–0.39 | Fair Quality (Usable Item) | |
0.20–0.29 | Potentially Poor Item (Consider Revising) | |
≤ 0.20 | Potentially Very Poor (Possibly Revise Substantially or Discard) | |
Bhat and Prasad [57] | > 0.35 | Excellent |
0.2–0.35 | Good | |
< 0.2 | Poor | |
Sugianto [6] | >0.40 | Very good |
0.30–0.39 | Reasonably good possibly need to improvement | |
0.20–0.29 | Marginal item usually needing and being to improvement | |
<0.19 | Poor item rejected or improved by revision | |
Aljehani, Pullishery et al. [66] and Sharma [4] | ≥ 0.40 | Very discriminating, very good item(Keep) |
0.30–0.39 | Discriminating item, good item (Keep) | |
0.20–0.29 | Moderately discriminating, fair item (Keep) | |
< 0.20 | Not discriminating item, marginal item (Revise/Discard) | |
Negative | Worst/ defective item (Definitely Discard) | |
Ramzan, Imran et al. [63] | > 0.30 | Excellent discrimination |
0.20–0.29 | Good discrimination | |
0–0.19 | Poor discrimination | |
00 | Defective | |
Uddin et al. [50] | ≥ 0.35 | Excellent |
0.25–0.34 | Good | |
0.21–0.24 | Acceptable | |
< 0.20 | Poor |
Reference values and interpretation of discrimination index (power).
When interpreting the discrimination power of an item to decide about, especial consideration should be related to its difficulty. Items with a high difficulty index (most of the examinee answer it right) and those with low difficulty index (most of the examinee answer it wrong) commonly have low discrimination power [35, 63]. In both cases, such items will not discriminate examines as the majority are on one side. Thus items with a moderate difficulty index are more likely to have good discrimination power.
The common causes of poor discrimination power of item include technical or writing flaws, untaught or not well covered content material, ambiguous wording, gray areas of opinion and controversy, and wrong keys [12, 50, 62, 66].
In general, the statistical data obtained from item analysis can help item constructors and exam composers to detect defective items. The decision to revise an item or distractors must be based on the difficulty index, discrimination index, and distractor efficiency. Revision of items can lead to modification in the teaching method or the content material [68].
Standard item analysis of mid-course examination. The total number of items is 40, and the total number of the examinee is 21. The KR20 is 0.82. Pt.Biserial: Point biserial correlation, Disc Index: discrimination index, Correct: number and percentage of the correct answer (difficulty index), Pct. Incorrect: percentage of an incorrect answer.
In this Example 1.
The number of examinees was 21.
The number of test items (Total possible) is 40.
The highest and lowest scores were 38 and 14 respectively.
The class average (mean) (30.3) is more than the class median (30) which represents a positively skewed distribution of examinee scores. Despite this, examinee scores may show normal ball-shape distribution. If the median is larger than the average (mean), the examinee scores will be negatively skewed distribution. Average equals median, the examinees’ scores are symmetrically (zero skewed) and normally distributed with ball-shaped.
The KR20 (Cronbach’s alpha) is 0.82 which is an acceptable value for most of the authors. Such value of internal consistency of exam allows deciding pass/fail. Lower values put the exam in questionable status.
Item 1: the difficulty index is 85.7% (easy). Although it has high discrimination power (DE = 0.6, Pbiserial = 0.58), two distractors are non-functional (B, C).
Comment: the item needs reediting. Distractors B and C need to be revised or changed by more plausible ones before being re-used.
Item 2: the difficulty index is 100% (easy). It has low discrimination power (DE = 00, Pbiserial = 00), all distractors are non-functional.
Comment: the item needs major revision or rewriting. This item is absolutely easy with no difficulty or discrimination index. Such items should be removed from the question bank and removal from the exam is considered valid.
Item 6: the difficulty index is 66.7% (moderate). It has high discrimination power (DE = 0.6, Pbiserial = 0.43) and all the distractors are functional.
Comment: The item has acceptable indices. Such items can be saved in the question bank for further use. The distractors need to be updated to have more efficiency.
Item 7: the difficulty index is 28.6% (difficult). Although it has high discrimination power (DE = 0.67, Pbiserial = 0.39), all the distractors are functional.
Comment: The item has acceptable indices. Such items can be saved in the question bank for further use. The distractors need to be updated to have more efficiency.
Item 8: the difficulty index is 76.2% (moderate). This item has a negative discrimination index (−0.33) and poor Pbiserial (0.04). Only one distractor is functional (C). The negative discrimination index is caused by the increased number of students in the lower account (27%) than those in the upper account (27%).
Comment: although the item has a moderate difficulty index, but is poorly discriminating. Such an item needs major revision.
Standard item analysis of Mid-course examination. The total number of items is 40, and the total number of examinee is 25. The KR20 is 0.74. Pt.Biserial: Point biserial correlation, Disc Index: discrimination index, Correct: number and percentage of the correct answer (difficulty index), Pct. Incorrect: percentage of an incorrect answer.
In this Example 2.
The number of examinees was 25.
The number of test items is 40.
The highest and lowest scores were 33 and 13 respectively.
The class average (mean) (24.6) is more than the class median (25), distribution of examinee scores is skewed to the left. Despite this, examinee scores may show normal ball shape distribution.
The KR20 (Cronbach’s alpha) is 0.74 which is an acceptable value for most of the authors. Such a value of internal consistency is suitable for class tests.
Item 8: the difficulty index is 4.0% (difficult). It has negative discrimination power (DE = -0.17, Pbiserial = −0.06), one distractors is non-functional (C).
Comment: the correct answer is (A) while most of the examinees chose (B). According to distractor analysis, this item is miss-keyed rather than an implausible distractor.
Item 9: the difficulty index is 20% (difficult). It has low discrimination power (DE = 0.17, Pbiserial = 0.09), all distractors are functional.
Comment: distractor analysis show option number (A) and (B) are more selected by examinees. This can be due to implausible. The presence of implausible can affect the item difficulty index. Distractors in this item should be revised or changed with plausible ones.
Item 11: the difficulty index is 44.0% (moderate). It has low discrimination power (DE = 0.0, Pbiserial = 0.01) and only one the distractors is non-functional.
Comment: The item has an acceptable difficulty index. Distractor (D) is more selected by upper examinee such as the key answer. Such a situation can favor missed key or implausible distractors. The distractors need to be updated to have more efficiency.
Book - collection of Works distributed in a book format, whose selection, coordination, preparation, and arrangement has been performed and published by IntechOpen, and in which the Work is included in its entirety in an unmodified form along with one or more other contributions, each constituting separate and independent sections, but together assembled into a collective whole.
",metaTitle:"Attribution Policy",metaDescription:"DEFINITION OF TERMS",metaKeywords:null,canonicalURL:"/page/attribution-policy",contentRaw:'[{"type":"htmlEditorComponent","content":"Work - a book Chapter (as well as Conference Papers), including any and all content, graphics, images and/or other materials forming part of, or accompanying, the Chapter/Conference Paper.
\\n\\nAttribution – appropriate credit for the used Work or book.
\\n\\nCreative Commons licenses – enable licensors to retain copyright while allowing others to use their Works in an appropriate way.
\\n\\nWith the purpose of protecting Authors' copyright and the transparent reuse of OA (Open Access) content, IntechOpen has developed Rules of Attribution of Works licensed under Creative Commons licenses.
\\n\\nIn case you reuse or republish any of the Works licensed under CC licenses, you must abide by the guidelines outlined below:
\\n\\nAll rights to Books and other compilations published on the IntechOpen platform and in print are reserved by IntechOpen. The Copyright to Books and other compilations is subject to a separate Copyright from any that exists in the included Works.
\\n\\nA Book in its entirety or a significant part of a Book cannot be translated freely without specific written consent by the publisher. Further information can be obtained at permissions@intechopen.com.
\\n\\nIn instances where permission is obtained from the publisher for reusing or republishing the Book, or significant parts of the Book, all of the following conditions apply:
\\n\\nEvery single Work that is used has to be attributed in the way described. If you are unsure about proper attribution, please write to permissions@intechopen.com.
\\n\\nIndividual Works originally published in IntechOpen books are licensed under Creative Commons licenses and can be freely used under terms of the respective CC license, if properly attributed. In order to properly attribute the Work you must respect all the conditions outlined below:
\\n\\nEvery single Work that is used has to be attributed in the way as described. If you are unsure about proper attribution, please contact Us at permissions@intechopen.com.
\\n\\nIn the event that you use more than one of IntechOpen's Works published in one or more books (but not a significant part of the book that is under separate Copyright), each of these have to be properly attributed in the way described.
\\n\\nIntechOpen does not have any claims on newly created copyrighted Works, but the Works originally published by IntechOpen must be properly attributed.
\\n\\nAll these rules apply to BOTH online and offline use.
\\n\\nParts of the Rules of Attribution are based on Work Attributing Creative Commons Materials published by the Australian Research Council Centre of Excellence for Creative Industries and Innovation, in partnership with Creative Commons Australia, which can be found at creativecommons.org.au licensed under Creative Commons Attribution 2.5 Australia license, and Best practices for attribution published by Creative Commons, which can be found at wiki.creativecommons.org under the Creative Commons Attribution 4.0 license.
\\n\\nAll the above rules are subject to change, IntechOpen reserves the right to take appropriate action if any of the conditions outlined above are not met.
\\n\\nPolicy last updated: 2016-06-09
\\n"}]'},components:[{type:"htmlEditorComponent",content:'Work - a book Chapter (as well as Conference Papers), including any and all content, graphics, images and/or other materials forming part of, or accompanying, the Chapter/Conference Paper.
\n\nAttribution – appropriate credit for the used Work or book.
\n\nCreative Commons licenses – enable licensors to retain copyright while allowing others to use their Works in an appropriate way.
\n\nWith the purpose of protecting Authors' copyright and the transparent reuse of OA (Open Access) content, IntechOpen has developed Rules of Attribution of Works licensed under Creative Commons licenses.
\n\nIn case you reuse or republish any of the Works licensed under CC licenses, you must abide by the guidelines outlined below:
\n\nAll rights to Books and other compilations published on the IntechOpen platform and in print are reserved by IntechOpen. The Copyright to Books and other compilations is subject to a separate Copyright from any that exists in the included Works.
\n\nA Book in its entirety or a significant part of a Book cannot be translated freely without specific written consent by the publisher. Further information can be obtained at permissions@intechopen.com.
\n\nIn instances where permission is obtained from the publisher for reusing or republishing the Book, or significant parts of the Book, all of the following conditions apply:
\n\nEvery single Work that is used has to be attributed in the way described. If you are unsure about proper attribution, please write to permissions@intechopen.com.
\n\nIndividual Works originally published in IntechOpen books are licensed under Creative Commons licenses and can be freely used under terms of the respective CC license, if properly attributed. In order to properly attribute the Work you must respect all the conditions outlined below:
\n\nEvery single Work that is used has to be attributed in the way as described. If you are unsure about proper attribution, please contact Us at permissions@intechopen.com.
\n\nIn the event that you use more than one of IntechOpen's Works published in one or more books (but not a significant part of the book that is under separate Copyright), each of these have to be properly attributed in the way described.
\n\nIntechOpen does not have any claims on newly created copyrighted Works, but the Works originally published by IntechOpen must be properly attributed.
\n\nAll these rules apply to BOTH online and offline use.
\n\nParts of the Rules of Attribution are based on Work Attributing Creative Commons Materials published by the Australian Research Council Centre of Excellence for Creative Industries and Innovation, in partnership with Creative Commons Australia, which can be found at creativecommons.org.au licensed under Creative Commons Attribution 2.5 Australia license, and Best practices for attribution published by Creative Commons, which can be found at wiki.creativecommons.org under the Creative Commons Attribution 4.0 license.
\n\nAll the above rules are subject to change, IntechOpen reserves the right to take appropriate action if any of the conditions outlined above are not met.
\n\nPolicy last updated: 2016-06-09
\n'}]},successStories:{items:[]},authorsAndEditors:{filterParams:{},profiles:[{id:"58592",title:"Dr.",name:"Arun",middleName:null,surname:"Shanker",slug:"arun-shanker",fullName:"Arun Shanker",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/58592/images/1664_n.jpg",biography:"Arun K. Shanker is serving as a Principal Scientist (Plant Physiology) with the Indian Council of Agricultural Research (ICAR) at the Central Research Institute for Dryland Agriculture in Hyderabad, India. He is working with the ICAR as a full time researcher since 1993 and has since earned his Advanced degree in Crop Physiology while in service. He has been awarded the prestigious Member of the Royal Society of Chemistry (MRSC), by the Royal Society of Chemistry, London in 2015. Presently he is working on systems biology approach to study the mechanism of abiotic stress tolerance in crops. His main focus now is to unravel the mechanism of drought and heat stress response in plants to tackle climate change related threats in agriculture.",institutionString:null,institution:{name:"Indian Council of Agricultural Research",country:{name:"India"}}},{id:"4782",title:"Prof.",name:"Bishnu",middleName:"P",surname:"Pal",slug:"bishnu-pal",fullName:"Bishnu Pal",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/4782/images/system/4782.jpg",biography:"Bishnu P. Pal is Professor of Physics at Mahindra École\nCentrale Hyderabad India since July 1st 2014 after retirement\nas Professor of Physics from IIT Delhi; Ph.D.’1975 from IIT\nDelhi; Fellow of OSA and SPIE; Senior Member IEEE;\nHonorary Foreign Member Royal Norwegian Society for\nScience and Arts; Member OSA Board of Directors (2009-\n11); Distinguished Lecturer IEEE Photonics Society (2005-\n07).",institutionString:null,institution:{name:"Indian Institute of Technology Delhi",country:{name:"India"}}},{id:"69653",title:"Dr.",name:"Chusak",middleName:null,surname:"Limsakul",slug:"chusak-limsakul",fullName:"Chusak Limsakul",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Prince of Songkla University",country:{name:"Thailand"}}},{id:"23804",title:"Dr.",name:"Hamzah",middleName:null,surname:"Arof",slug:"hamzah-arof",fullName:"Hamzah Arof",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/23804/images/5492_n.jpg",biography:"Hamzah Arof received his BSc from Michigan State University, and PhD from the University of Wales. Both degrees were in electrical engineering. His current research interests include signal processing and photonics. Currently he is affiliated with the Department of Electrical Engineering, University of Malaya, Malaysia.",institutionString:null,institution:{name:"University of Malaya",country:{name:"Malaysia"}}},{id:"41989",title:"Prof.",name:"He",middleName:null,surname:"Tian",slug:"he-tian",fullName:"He Tian",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"East China University of Science and Technology",country:{name:"China"}}},{id:"33351",title:null,name:"Hendra",middleName:null,surname:"Hermawan",slug:"hendra-hermawan",fullName:"Hendra Hermawan",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/33351/images/168_n.jpg",biography:null,institutionString:null,institution:{name:"Institut Teknologi Bandung",country:{name:"Indonesia"}}},{id:"11981",title:"Prof.",name:"Hiroshi",middleName:null,surname:"Ishiguro",slug:"hiroshi-ishiguro",fullName:"Hiroshi Ishiguro",position:null,profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002bRglaQAC/Profile_Picture_1626411846553",biography:"Hiroshi Ishiguro is an award-winning roboticist and innovator. As the Director of the Intelligent Robotics Laboratory, which is part of the Department of Systems Innovation in the Graduate School of Engineering Science at Osaka University, Japan, Ishiguro concentrates on making robots that are similar as possible to humans to understand the human species. A notable project of his laboratory is the Actroid, a humanoid robot with a lifelike appearance and observable behavior such as facial movements. (Sources: http://www.geminoid.jp/en/index.html, https://en.wikipedia.org/wiki/Hiroshi_Ishiguro)",institutionString:null,institution:{name:"Osaka University",country:{name:"Japan"}}},{id:"45747",title:"Dr.",name:"Hsin-I",middleName:null,surname:"Chang",slug:"hsin-i-chang",fullName:"Hsin-I Chang",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"National Chiayi University",country:{name:"Taiwan"}}},{id:"61581",title:"Dr.",name:"Joy Rizki Pangestu",middleName:null,surname:"Djuansjah",slug:"joy-rizki-pangestu-djuansjah",fullName:"Joy Rizki Pangestu Djuansjah",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/61581/images/237_n.jpg",biography:null,institutionString:null,institution:{name:"University of Technology Malaysia",country:{name:"Malaysia"}}},{id:"94249",title:"Prof.",name:"Junji",middleName:null,surname:"Kido",slug:"junji-kido",fullName:"Junji Kido",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Yamagata University",country:{name:"Japan"}}},{id:"12009",title:"Dr.",name:"Ki Young",middleName:null,surname:"Kim",slug:"ki-young-kim",fullName:"Ki Young Kim",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/12009/images/system/12009.jpg",biography:"Http://m80.knu.ac.kr/~doors",institutionString:null,institution:{name:"National Cheng Kung University",country:{name:"Taiwan"}}},{id:"132595",title:"Prof.",name:"Long",middleName:null,surname:"Wang",slug:"long-wang",fullName:"Long Wang",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Peking University",country:{name:"China"}}}],filtersByRegion:[{group:"region",caption:"North America",value:1,count:6583},{group:"region",caption:"Middle and South America",value:2,count:5888},{group:"region",caption:"Africa",value:3,count:2381},{group:"region",caption:"Asia",value:4,count:12511},{group:"region",caption:"Australia and Oceania",value:5,count:1006},{group:"region",caption:"Europe",value:6,count:17529}],offset:12,limit:12,total:12514},chapterEmbeded:{data:{}},editorApplication:{success:null,errors:{}},ofsBooks:{filterParams:{topicId:"156"},books:[],filtersByTopic:[{group:"topic",caption:"Agricultural and Biological Sciences",value:5,count:43},{group:"topic",caption:"Biochemistry, Genetics and Molecular Biology",value:6,count:11},{group:"topic",caption:"Business, Management and Economics",value:7,count:6},{group:"topic",caption:"Chemistry",value:8,count:21},{group:"topic",caption:"Computer and Information Science",value:9,count:21},{group:"topic",caption:"Earth and Planetary Sciences",value:10,count:17},{group:"topic",caption:"Engineering",value:11,count:59},{group:"topic",caption:"Environmental Sciences",value:12,count:8},{group:"topic",caption:"Immunology and Microbiology",value:13,count:10},{group:"topic",caption:"Materials Science",value:14,count:27},{group:"topic",caption:"Mathematics",value:15,count:9},{group:"topic",caption:"Medicine",value:16,count:122},{group:"topic",caption:"Nanotechnology and Nanomaterials",value:17,count:9},{group:"topic",caption:"Neuroscience",value:18,count:3},{group:"topic",caption:"Pharmacology, Toxicology and Pharmaceutical Science",value:19,count:7},{group:"topic",caption:"Physics",value:20,count:11},{group:"topic",caption:"Psychology",value:21,count:10},{group:"topic",caption:"Robotics",value:22,count:4},{group:"topic",caption:"Social Sciences",value:23,count:8},{group:"topic",caption:"Veterinary Medicine and Science",value:25,count:4}],offset:12,limit:12,total:0},popularBooks:{featuredBooks:[{type:"book",id:"10584",title:"Engineered Wood Products for Construction",subtitle:null,isOpenForSubmission:!1,hash:"421757c56a3735986055250821275a51",slug:"engineered-wood-products-for-construction",bookSignature:"Meng Gong",coverURL:"https://cdn.intechopen.com/books/images_new/10584.jpg",editors:[{id:"274242",title:"Dr.",name:"Meng",middleName:null,surname:"Gong",slug:"meng-gong",fullName:"Meng Gong"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"10222",title:"Demyelination Disorders",subtitle:null,isOpenForSubmission:!1,hash:"b6c26ceccacdde70c41c587361bd5558",slug:"demyelination-disorders",bookSignature:"Stavros J. Baloyannis, Fabian H. Rossi and Welwin Liu",coverURL:"https://cdn.intechopen.com/books/images_new/10222.jpg",editors:[{id:"156098",title:"Emeritus Prof.",name:"Stavros J.",middleName:"J.",surname:"Baloyannis",slug:"stavros-j.-baloyannis",fullName:"Stavros J. Baloyannis"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"9544",title:"Global Trade in the Emerging Business Environment",subtitle:null,isOpenForSubmission:!1,hash:"fb8cb09b9599246add78d508a98273d5",slug:"global-trade-in-the-emerging-business-environment",bookSignature:"Muhammad Mohiuddin, Jingbin Wang , Md. Samim Al Azad and Selim Ahmed",coverURL:"https://cdn.intechopen.com/books/images_new/9544.jpg",editors:[{id:"418514",title:"Dr.",name:"Muhammad",middleName:null,surname:"Mohiuddin",slug:"muhammad-mohiuddin",fullName:"Muhammad Mohiuddin"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"10979",title:"Parenting",subtitle:"Challenges of Child Rearing in a Changing Society",isOpenForSubmission:!1,hash:"6f345ebcf4fd61e73643c69063a12c7b",slug:"parenting-challenges-of-child-rearing-in-a-changing-society",bookSignature:"Sayyed Ali Samadi",coverURL:"https://cdn.intechopen.com/books/images_new/10979.jpg",editors:[{id:"52145",title:"Dr.",name:"Sayyed Ali",middleName:null,surname:"Samadi",slug:"sayyed-ali-samadi",fullName:"Sayyed Ali Samadi"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"9808",title:"Contemporary Topics in Patient Safety",subtitle:"Volume 1",isOpenForSubmission:!1,hash:"fb6371607c2c6c02c6a2af8892765aba",slug:"contemporary-topics-in-patient-safety-volume-1",bookSignature:"Stanislaw P. Stawicki and Michael S. Firstenberg",coverURL:"https://cdn.intechopen.com/books/images_new/9808.jpg",editors:[{id:"181694",title:"Dr.",name:"Stanislaw P.",middleName:null,surname:"Stawicki",slug:"stanislaw-p.-stawicki",fullName:"Stanislaw P. Stawicki"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"10681",title:"Biodegradation Technology of Organic and Inorganic Pollutants",subtitle:null,isOpenForSubmission:!1,hash:"9a6e10e02788092872fd249436898e97",slug:"biodegradation-technology-of-organic-and-inorganic-pollutants",bookSignature:"Kassio Ferreira Mendes, Rodrigo Nogueira de Sousa and Kamila Cabral Mielke",coverURL:"https://cdn.intechopen.com/books/images_new/10681.jpg",editors:[{id:"197720",title:"Ph.D.",name:"Kassio",middleName:null,surname:"Ferreira Mendes",slug:"kassio-ferreira-mendes",fullName:"Kassio Ferreira Mendes"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"10764",title:"Antenna Systems",subtitle:null,isOpenForSubmission:!1,hash:"2fbf1c7a5d92723f08198fc9b526a8ad",slug:"antenna-systems",bookSignature:"Hussain Al-Rizzo and Said Abushamleh",coverURL:"https://cdn.intechopen.com/books/images_new/10764.jpg",editors:[{id:"153384",title:"Prof.",name:"Hussain",middleName:null,surname:"Al-Rizzo",slug:"hussain-al-rizzo",fullName:"Hussain Al-Rizzo"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"10668",title:"Sustainability of Concrete With Synthetic and Recycled Aggregates",subtitle:null,isOpenForSubmission:!1,hash:"55856c6a8bc3a5b21dae5a1af09a56b6",slug:"sustainability-of-concrete-with-synthetic-and-recycled-aggregates",bookSignature:"Hosam M. Saleh",coverURL:"https://cdn.intechopen.com/books/images_new/10668.jpg",editors:[{id:"144691",title:"Prof.",name:"Hosam",middleName:null,surname:"Saleh",slug:"hosam-saleh",fullName:"Hosam Saleh"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"10803",title:"Reactive Oxygen Species",subtitle:null,isOpenForSubmission:!1,hash:"176adcf090fdd1f93cb8ce3146e79ca1",slug:"reactive-oxygen-species",bookSignature:"Rizwan Ahmad",coverURL:"https://cdn.intechopen.com/books/images_new/10803.jpg",editors:[{id:"40482",title:null,name:"Rizwan",middleName:null,surname:"Ahmad",slug:"rizwan-ahmad",fullName:"Rizwan Ahmad"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"9032",title:"Corporate Social Responsibility",subtitle:null,isOpenForSubmission:!1,hash:"f609bf3251d7cc7bae0099a4374adfc3",slug:"corporate-social-responsibility",bookSignature:"Beatrice Orlando",coverURL:"https://cdn.intechopen.com/books/images_new/9032.jpg",editors:[{id:"232969",title:"Prof.",name:"Beatrice",middleName:null,surname:"Orlando",slug:"beatrice-orlando",fullName:"Beatrice Orlando"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"10351",title:"Enhanced Liposuction",subtitle:"New Perspectives and Techniques",isOpenForSubmission:!1,hash:"f08ed6de16da357614586c5b58ed4dfa",slug:"enhanced-liposuction-new-perspectives-and-techniques",bookSignature:"Diane Irvine Duncan",coverURL:"https://cdn.intechopen.com/books/images_new/10351.jpg",editors:[{id:"279869",title:"Dr.",name:"Diane Irvine",middleName:null,surname:"Duncan",slug:"diane-irvine-duncan",fullName:"Diane Irvine Duncan"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"10779",title:"21st Century Nanostructured Materials",subtitle:"Physics, Chemistry, Classification, and Emerging Applications in Industry, Biomedicine, and Agriculture",isOpenForSubmission:!1,hash:"72c67f97f9bef68200df115b5fd79884",slug:"21st-century-nanostructured-materials-physics-chemistry-classification-and-emerging-applications-in-industry-biomedicine-and-agriculture",bookSignature:"Phuong V. Pham",coverURL:"https://cdn.intechopen.com/books/images_new/10779.jpg",editors:[{id:"236073",title:"Dr.",name:"Phuong",middleName:"Viet",surname:"Pham",slug:"phuong-pham",fullName:"Phuong Pham"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter"}}],offset:12,limit:12,total:4386},hotBookTopics:{hotBooks:[],offset:0,limit:12,total:null},publish:{},publishingProposal:{success:null,errors:{}},books:{featuredBooks:[{type:"book",id:"10584",title:"Engineered Wood Products for Construction",subtitle:null,isOpenForSubmission:!1,hash:"421757c56a3735986055250821275a51",slug:"engineered-wood-products-for-construction",bookSignature:"Meng Gong",coverURL:"https://cdn.intechopen.com/books/images_new/10584.jpg",publishedDate:"April 28th 2022",numberOfDownloads:3665,editors:[{id:"274242",title:"Dr.",name:"Meng",middleName:null,surname:"Gong",slug:"meng-gong",fullName:"Meng Gong"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"10222",title:"Demyelination Disorders",subtitle:null,isOpenForSubmission:!1,hash:"b6c26ceccacdde70c41c587361bd5558",slug:"demyelination-disorders",bookSignature:"Stavros J. Baloyannis, Fabian H. Rossi and Welwin Liu",coverURL:"https://cdn.intechopen.com/books/images_new/10222.jpg",publishedDate:"May 4th 2022",numberOfDownloads:1713,editors:[{id:"156098",title:"Emeritus Prof.",name:"Stavros J.",middleName:"J.",surname:"Baloyannis",slug:"stavros-j.-baloyannis",fullName:"Stavros J. Baloyannis"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"9544",title:"Global Trade in the Emerging Business Environment",subtitle:null,isOpenForSubmission:!1,hash:"fb8cb09b9599246add78d508a98273d5",slug:"global-trade-in-the-emerging-business-environment",bookSignature:"Muhammad Mohiuddin, Jingbin Wang , Md. Samim Al Azad and Selim Ahmed",coverURL:"https://cdn.intechopen.com/books/images_new/9544.jpg",publishedDate:"April 28th 2022",numberOfDownloads:2481,editors:[{id:"418514",title:"Dr.",name:"Muhammad",middleName:null,surname:"Mohiuddin",slug:"muhammad-mohiuddin",fullName:"Muhammad Mohiuddin"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"10979",title:"Parenting",subtitle:"Challenges of Child Rearing in a Changing Society",isOpenForSubmission:!1,hash:"6f345ebcf4fd61e73643c69063a12c7b",slug:"parenting-challenges-of-child-rearing-in-a-changing-society",bookSignature:"Sayyed Ali Samadi",coverURL:"https://cdn.intechopen.com/books/images_new/10979.jpg",publishedDate:"May 4th 2022",numberOfDownloads:1107,editors:[{id:"52145",title:"Dr.",name:"Sayyed Ali",middleName:null,surname:"Samadi",slug:"sayyed-ali-samadi",fullName:"Sayyed Ali Samadi"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"9808",title:"Contemporary Topics in Patient Safety",subtitle:"Volume 1",isOpenForSubmission:!1,hash:"fb6371607c2c6c02c6a2af8892765aba",slug:"contemporary-topics-in-patient-safety-volume-1",bookSignature:"Stanislaw P. Stawicki and Michael S. Firstenberg",coverURL:"https://cdn.intechopen.com/books/images_new/9808.jpg",publishedDate:"April 20th 2022",numberOfDownloads:3307,editors:[{id:"181694",title:"Dr.",name:"Stanislaw P.",middleName:null,surname:"Stawicki",slug:"stanislaw-p.-stawicki",fullName:"Stanislaw P. Stawicki"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"10681",title:"Biodegradation Technology of Organic and Inorganic Pollutants",subtitle:null,isOpenForSubmission:!1,hash:"9a6e10e02788092872fd249436898e97",slug:"biodegradation-technology-of-organic-and-inorganic-pollutants",bookSignature:"Kassio Ferreira Mendes, Rodrigo Nogueira de Sousa and Kamila Cabral Mielke",coverURL:"https://cdn.intechopen.com/books/images_new/10681.jpg",publishedDate:"April 20th 2022",numberOfDownloads:3266,editors:[{id:"197720",title:"Ph.D.",name:"Kassio",middleName:null,surname:"Ferreira Mendes",slug:"kassio-ferreira-mendes",fullName:"Kassio Ferreira Mendes"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"10764",title:"Antenna Systems",subtitle:null,isOpenForSubmission:!1,hash:"2fbf1c7a5d92723f08198fc9b526a8ad",slug:"antenna-systems",bookSignature:"Hussain Al-Rizzo and Said Abushamleh",coverURL:"https://cdn.intechopen.com/books/images_new/10764.jpg",publishedDate:"April 28th 2022",numberOfDownloads:1868,editors:[{id:"153384",title:"Prof.",name:"Hussain",middleName:null,surname:"Al-Rizzo",slug:"hussain-al-rizzo",fullName:"Hussain Al-Rizzo"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"10668",title:"Sustainability of Concrete With Synthetic and Recycled Aggregates",subtitle:null,isOpenForSubmission:!1,hash:"55856c6a8bc3a5b21dae5a1af09a56b6",slug:"sustainability-of-concrete-with-synthetic-and-recycled-aggregates",bookSignature:"Hosam M. Saleh",coverURL:"https://cdn.intechopen.com/books/images_new/10668.jpg",publishedDate:"May 4th 2022",numberOfDownloads:856,editors:[{id:"144691",title:"Prof.",name:"Hosam",middleName:null,surname:"Saleh",slug:"hosam-saleh",fullName:"Hosam Saleh"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"10803",title:"Reactive Oxygen Species",subtitle:null,isOpenForSubmission:!1,hash:"176adcf090fdd1f93cb8ce3146e79ca1",slug:"reactive-oxygen-species",bookSignature:"Rizwan Ahmad",coverURL:"https://cdn.intechopen.com/books/images_new/10803.jpg",publishedDate:"April 28th 2022",numberOfDownloads:1704,editors:[{id:"40482",title:null,name:"Rizwan",middleName:null,surname:"Ahmad",slug:"rizwan-ahmad",fullName:"Rizwan Ahmad"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"9032",title:"Corporate Social Responsibility",subtitle:null,isOpenForSubmission:!1,hash:"f609bf3251d7cc7bae0099a4374adfc3",slug:"corporate-social-responsibility",bookSignature:"Beatrice Orlando",coverURL:"https://cdn.intechopen.com/books/images_new/9032.jpg",publishedDate:"March 16th 2022",numberOfDownloads:7489,editors:[{id:"232969",title:"Prof.",name:"Beatrice",middleName:null,surname:"Orlando",slug:"beatrice-orlando",fullName:"Beatrice Orlando"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter"}}],latestBooks:[{type:"book",id:"10251",title:"Plankton Communities",subtitle:null,isOpenForSubmission:!1,hash:"e11e441ca2d2d5f631b1b4704505cfb6",slug:"plankton-communities",bookSignature:"Leonel Pereira and Ana Marta Gonçalves",coverURL:"https://cdn.intechopen.com/books/images_new/10251.jpg",editedByType:"Edited by",publishedDate:"May 18th 2022",editors:[{id:"279788",title:"Dr.",name:"Leonel",middleName:null,surname:"Pereira",slug:"leonel-pereira",fullName:"Leonel Pereira"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"10830",title:"Animal Feed Science and Nutrition",subtitle:"Production, Health and Environment",isOpenForSubmission:!1,hash:"79944fc8fbbaa329aed6fde388154832",slug:"animal-feed-science-and-nutrition-production-health-and-environment",bookSignature:"Amlan Kumar Patra",coverURL:"https://cdn.intechopen.com/books/images_new/10830.jpg",editedByType:"Edited by",publishedDate:"May 18th 2022",editors:[{id:"310962",title:"Dr.",name:"Amlan",middleName:"Kumar",surname:"Patra",slug:"amlan-patra",fullName:"Amlan Patra"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"10853",title:"Recent Advances in Polynomials",subtitle:null,isOpenForSubmission:!1,hash:"9e8671bae09ccaa8b8e276c639a737fc",slug:"recent-advances-in-polynomials",bookSignature:"Kamal Shah",coverURL:"https://cdn.intechopen.com/books/images_new/10853.jpg",editedByType:"Edited by",publishedDate:"May 18th 2022",editors:[{id:"231748",title:"Dr.",name:"Kamal",middleName:null,surname:"Shah",slug:"kamal-shah",fullName:"Kamal Shah"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"10871",title:"Computed-Tomography (CT) Scan",subtitle:null,isOpenForSubmission:!1,hash:"966d8cf74fa27eea1b9cbc9a6ee94993",slug:"computed-tomography-ct-scan",bookSignature:"Reda R. Gharieb",coverURL:"https://cdn.intechopen.com/books/images_new/10871.jpg",editedByType:"Edited by",publishedDate:"May 18th 2022",editors:[{id:"225387",title:"Prof.",name:"Reda R.",middleName:"R.",surname:"Gharieb",slug:"reda-r.-gharieb",fullName:"Reda R. Gharieb"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"10903",title:"Genetically Modified Plants and Beyond",subtitle:null,isOpenForSubmission:!1,hash:"4d7ed4faab99c92cd4d676dc86501df9",slug:"genetically-modified-plants-and-beyond",bookSignature:"Idah Sithole Niang",coverURL:"https://cdn.intechopen.com/books/images_new/10903.jpg",editedByType:"Edited by",publishedDate:"May 18th 2022",editors:[{id:"90172",title:"Prof.",name:"Idah",middleName:null,surname:"Sithole-Niang",slug:"idah-sithole-niang",fullName:"Idah Sithole-Niang"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"10904",title:"Fusarium",subtitle:"An Overview of the Genus",isOpenForSubmission:!1,hash:"49d9063e43f94bd1517d65fbc58b93c3",slug:"fusarium-an-overview-of-the-genus",bookSignature:"Seyed Mahyar Mirmajlessi",coverURL:"https://cdn.intechopen.com/books/images_new/10904.jpg",editedByType:"Edited by",publishedDate:"May 18th 2022",editors:[{id:"100573",title:"Dr.",name:"Seyed Mahyar",middleName:null,surname:"Mirmajlessi",slug:"seyed-mahyar-mirmajlessi",fullName:"Seyed Mahyar Mirmajlessi"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"10654",title:"Brain-Computer Interface",subtitle:null,isOpenForSubmission:!1,hash:"a5308884068cc53ed31c6baba756857f",slug:"brain-computer-interface",bookSignature:"Vahid Asadpour",coverURL:"https://cdn.intechopen.com/books/images_new/10654.jpg",editedByType:"Edited by",publishedDate:"May 18th 2022",editors:[{id:"165328",title:"Dr.",name:"Vahid",middleName:null,surname:"Asadpour",slug:"vahid-asadpour",fullName:"Vahid Asadpour"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"10676",title:"Recent Applications in Graph Theory",subtitle:null,isOpenForSubmission:!1,hash:"900c60742d224080732bd16bd25ccba8",slug:"recent-applications-in-graph-theory",bookSignature:"Harun Pirim",coverURL:"https://cdn.intechopen.com/books/images_new/10676.jpg",editedByType:"Edited by",publishedDate:"May 18th 2022",editors:[{id:"146092",title:"Dr.",name:"Harun",middleName:null,surname:"Pirim",slug:"harun-pirim",fullName:"Harun Pirim"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"11196",title:"New Updates in E-Learning",subtitle:null,isOpenForSubmission:!1,hash:"6afaadf68e2a0a4b370ac5ceb5ca89c6",slug:"new-updates-in-e-learning",bookSignature:"Eduard Babulak",coverURL:"https://cdn.intechopen.com/books/images_new/11196.jpg",editedByType:"Edited by",publishedDate:"May 18th 2022",editors:[{id:"10086",title:"Prof.",name:"Eduard",middleName:null,surname:"Babulak",slug:"eduard-babulak",fullName:"Eduard Babulak"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"9974",title:"E-Learning and Digital Education in the Twenty-First Century",subtitle:null,isOpenForSubmission:!1,hash:"88b58d66e975df20425fc1dfd22d53aa",slug:"e-learning-and-digital-education-in-the-twenty-first-century",bookSignature:"M. Mahruf C. Shohel",coverURL:"https://cdn.intechopen.com/books/images_new/9974.jpg",editedByType:"Edited by",publishedDate:"May 18th 2022",editors:[{id:"94099",title:"Dr.",name:"M. Mahruf C.",middleName:null,surname:"Shohel",slug:"m.-mahruf-c.-shohel",fullName:"M. Mahruf C. Shohel"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}}]},subject:{topic:{id:"785",title:"Geomatics",slug:"geomatics",parent:{id:"118",title:"Environmental Engineering",slug:"engineering-environmental-engineering"},numberOfBooks:2,numberOfSeries:0,numberOfAuthorsAndEditors:40,numberOfWosCitations:18,numberOfCrossrefCitations:13,numberOfDimensionsCitations:23,videoUrl:null,fallbackUrl:null,description:null},booksByTopicFilter:{topicId:"785",sort:"-publishedDate",limit:12,offset:0},booksByTopicCollection:[{type:"book",id:"7465",title:"Trends in Geomatics",subtitle:"An Earth Science Perspective",isOpenForSubmission:!1,hash:"a32ff40d6c47e2b545b92d5075508c9c",slug:"trends-in-geomatics-an-earth-science-perspective",bookSignature:"Rifaat Abdalla",coverURL:"https://cdn.intechopen.com/books/images_new/7465.jpg",editedByType:"Edited by",editors:[{id:"222877",title:"Dr.",name:"Rifaat",middleName:null,surname:"Abdalla",slug:"rifaat-abdalla",fullName:"Rifaat Abdalla"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"5246",title:"Geospatial Technology",subtitle:"Environmental and Social Applications",isOpenForSubmission:!1,hash:"79ea6b081cf0704d747c97857464d3fd",slug:"geospatial-technology-environmental-and-social-applications",bookSignature:"Pasquale Imperatore and Antonio Pepe",coverURL:"https://cdn.intechopen.com/books/images_new/5246.jpg",editedByType:"Edited by",editors:[{id:"4222",title:"Dr.",name:"Pasquale",middleName:null,surname:"Imperatore",slug:"pasquale-imperatore",fullName:"Pasquale Imperatore"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}}],booksByTopicTotal:2,seriesByTopicCollection:[],seriesByTopicTotal:0,mostCitedChapters:[{id:"51565",doi:"10.5772/64214",title:"Collaborative Uses of Geospatial Technology to Support Climate Change Adaptation in Indigenous Communities of the Circumpolar North",slug:"collaborative-uses-of-geospatial-technology-to-support-climate-change-adaptation-in-indigenous-commu",totalDownloads:1457,totalCrossrefCites:2,totalDimensionsCites:5,abstract:"A literature review is conducted of geospatial technologies in community-based research on ice and mobility among Indigenous people of the circumpolar north. Numerous studies explore the use of traditional knowledge in the Arctic on sea ice, but limited evidence of community-based research in sub-Arctic communities and in freshwater ice systems is found. Geographical Information Systems (GIS) and remote sensing tools have been applied in a variety of ways in support of community adaptations. These include the production of living memory maps, ice classification systems, and geodatabases that reflect the relationship-building nature of collaborations between Indigenous traditional knowledge holders and scientists. Satellite imagery—particularly synthetic aperture radar (SAR)—is widely used to characterize traditional understandings of ice to help tailor geospatial tools, climate research, and early warning systems, so that they may be used more effectively to address community interests and needs. As numerous mapping platforms have been developed in the circumpolar north, there are important considerations with respect to data management, Indigenous rights, and data sharing. We see opportunities for further research in lake and river ice, and in further developing early warning systems to address the growing problem of unpredictable ice regimes in Arctic and sub-Arctic regions.",book:{id:"5246",slug:"geospatial-technology-environmental-and-social-applications",title:"Geospatial Technology",fullTitle:"Geospatial Technology - Environmental and Social Applications"},signatures:"Megan Sheremata, Leonard J.S. Tsuji and William A. Gough",authors:[{id:"182233",title:"Ph.D. Student",name:"Megan",middleName:null,surname:"Sheremata",slug:"megan-sheremata",fullName:"Megan Sheremata"},{id:"182238",title:"Prof.",name:"William A.",middleName:null,surname:"Gough",slug:"william-a.-gough",fullName:"William A. Gough"},{id:"182241",title:"Prof.",name:"Leonard J. S.",middleName:null,surname:"Tsuji",slug:"leonard-j.-s.-tsuji",fullName:"Leonard J. S. Tsuji"}]},{id:"51625",doi:"10.5772/64303",title:"Participatory Mapping to Disrupt Unjust Urban Trajectories in Lima",slug:"participatory-mapping-to-disrupt-unjust-urban-trajectories-in-lima",totalDownloads:1643,totalCrossrefCites:3,totalDimensionsCites:4,abstract:"This chapter shares the experience of two action research projects ReMap Lima and cLIMA sin Riego, where mapping has been used with three main objectives: to make visible what is otherwise ‘invisible’; to open up dialogue between different stakeholders in the city and to arrive at concrete actions, collectively negotiated between citizens and policy makers. Two case study sites were chosen in Lima, Peru: Barrios Altos (BA) in the historic centre and José Carlos Mariátegui (JCM) at the edge of the city. The approach adopted applies a participatory action methodology based on grounded applications and advanced technologies for community-led mapping and visualisation. The chapter reflects upon three interrelated sites of the mapping process: the reading, writing and audiencing of maps and explores how these can provide opportunities to break away from the polar positions often established between Claimant/ marginalised group and the state, thus aiming to contribute to a process of spatial co-learning across typically confronted actors. The two case studies show different possibilities for interrogating the city to provide a spatially and socially grounded way of co-producing knowledge for action that can contribute to the planning of just urban futures.",book:{id:"5246",slug:"geospatial-technology-environmental-and-social-applications",title:"Geospatial Technology",fullTitle:"Geospatial Technology - Environmental and Social Applications"},signatures:"Rita Lambert and Adriana Allen",authors:[{id:"183462",title:"Dr.",name:"Adriana",middleName:null,surname:"Allen",slug:"adriana-allen",fullName:"Adriana Allen"},{id:"184062",title:"Dr.",name:"Rita",middleName:null,surname:"Lambert",slug:"rita-lambert",fullName:"Rita Lambert"}]},{id:"51416",doi:"10.5772/64250",title:"Satellite SAR Interferometry for Earth’s Crust Deformation Monitoring and Geological Phenomena Analysis",slug:"satellite-sar-interferometry-for-earth-s-crust-deformation-monitoring-and-geological-phenomena-analy",totalDownloads:1441,totalCrossrefCites:2,totalDimensionsCites:4,abstract:"Synthetic aperture radar interferometry (InSAR) and the related processing techniques provide a unique tool for the quantitative measurement of the Earth’s surface deformation associated with certain geophysical processes (such as volcanic eruptions, landslides and earthquakes), thus making possible long-term monitoring of surface deformation and analysis of relevant geodynamic phenomena. This chapter provides an application-oriented perspective on the spaceborne InSAR technology with emphasis on subsequent geophysical investigations. First, the fundamentals of radar interferometry and differential interferometry, as well as error sources, are briefly introduced. Emphasis is then placed on the realistic simulation of the underlying geophysics processes, thus offering an unfolded perspective on both analytical and numerical approaches for modeling deformation sources. Finally, various experimental investigations conducted by acquiring SAR multitemporal observations on areas subject to deformation processes of particular geological interest are presented and discussed.",book:{id:"5246",slug:"geospatial-technology-environmental-and-social-applications",title:"Geospatial Technology",fullTitle:"Geospatial Technology - Environmental and Social Applications"},signatures:"Giuseppe Solaro, Pasquale Imperatore and Antonio Pepe",authors:[{id:"4222",title:"Dr.",name:"Pasquale",middleName:null,surname:"Imperatore",slug:"pasquale-imperatore",fullName:"Pasquale Imperatore"},{id:"99269",title:"Dr.",name:"Antonio",middleName:null,surname:"Pepe",slug:"antonio-pepe",fullName:"Antonio Pepe"},{id:"182234",title:"Ph.D.",name:"Giuseppe",middleName:null,surname:"Solaro",slug:"giuseppe-solaro",fullName:"Giuseppe Solaro"}]},{id:"51731",doi:"10.5772/64527",title:"Estimation and Uncertainty Assessment of Surface Microclimate Indicators at Local Scale Using Airborne Infrared Thermography and Multispectral Imagery",slug:"estimation-and-uncertainty-assessment-of-surface-microclimate-indicators-at-local-scale-using-airbor",totalDownloads:1469,totalCrossrefCites:1,totalDimensionsCites:2,abstract:"A precise estimation and the characterization of the spatial variability of microclimate conditions (MCCs) are essential for risk assessment and site-specific management of vector-borne diseases and crop pests. The objective of this study was to estimate at local scale, and assess the uncertainties of Surface Microclimate Indicators (SMIs) derived from airborne infrared thermography and multispectral imaging. SMIs including Surface Temperature (ST) were estimated in southern Quebec, Canada. The formulation of their uncertainties was based on in-situ observations and the law of propagation of uncertainty. SMIs showed strong local variability and intra-plot variability of MCCs in the study area. The ST values ranged from 290 K to 331 K. They varied more than 17 K on vegetable crop fields. The correlation between ST and in-situ observations was very high (r = 0.99, p = 0.010). The uncertainty and the bias of ST compared to in-situ observations were 0.73 K and ±1.42 K respectively. This study demonstrated that very high spatial resolution multispectral imaging and infrared thermography present a good potential for the characterization of the MCCs that govern the abundance and the behavior of disease vectors and crop pests in a given area.",book:{id:"5246",slug:"geospatial-technology-environmental-and-social-applications",title:"Geospatial Technology",fullTitle:"Geospatial Technology - Environmental and Social Applications"},signatures:"Serge Olivier Kotchi, Nathalie Barrette, Alain A. Viau, Jae-Dong\nJang, Valéry Gond and Mir Abolfazl Mostafavi",authors:[{id:"31099",title:"Prof.",name:"Mir Abolfazl",middleName:null,surname:"Mostafavi",slug:"mir-abolfazl-mostafavi",fullName:"Mir Abolfazl Mostafavi"},{id:"181360",title:"Ph.D.",name:"Serge Olivier",middleName:null,surname:"Kotchi",slug:"serge-olivier-kotchi",fullName:"Serge Olivier Kotchi"},{id:"188549",title:"Dr.",name:"Nathalie",middleName:null,surname:"Barrette",slug:"nathalie-barrette",fullName:"Nathalie Barrette"},{id:"188550",title:"Dr.",name:"Alain A.",middleName:null,surname:"Viau",slug:"alain-a.-viau",fullName:"Alain A. Viau"},{id:"188551",title:"Dr.",name:"Jae-Dong",middleName:null,surname:"Jang",slug:"jae-dong-jang",fullName:"Jae-Dong Jang"},{id:"188552",title:"Dr.",name:"Valery",middleName:null,surname:"Gond",slug:"valery-gond",fullName:"Valery Gond"}]},{id:"51786",doi:"10.5772/64528",title:"GIS Applications in Agronomy",slug:"gis-applications-in-agronomy",totalDownloads:3022,totalCrossrefCites:1,totalDimensionsCites:2,abstract:"Agronomy is a branch of agriculture that deals with soil and crop. Soil varies in space and is responsible for variation in the growth and yield of crops on the field. This variation in the yields of crops planted and monitored on the same parcel of land under the same environmental conditions has been a great concern to farmers. Spatial variations of soil nutrients status, as caused by topography, soil texture and management practices, have been observed across the fields. Hence, the need to separate the field into site specific management units using geographical information systems (GIS) for effective soil and crop management in order to obtain optimum productivity. Over the years, field sizes, farming direction, locations of fences, rotations and fertility programmes have changed the nutritional status of the farms. Consequently, the productivity of the soil has equally been affected. In spite of these factors, conventional agriculture treats an entire field uniformly with respect to the application of fertiliser, pesticides, soil amendments and other chemical application. The use of GIS will help farmers to overcome over- or under-applications of fertiliser and other agrochemical applications. The potential of GIS application in agronomy is obviously large. However, the GIS user community in the field of agronomy is rather small compared to other business sectors. To advance the use of GIS in agronomic studies, this Chapter in book tends to explore the applications of GIS to some fields in agronomy.",book:{id:"5246",slug:"geospatial-technology-environmental-and-social-applications",title:"Geospatial Technology",fullTitle:"Geospatial Technology - Environmental and Social Applications"},signatures:"Suarau O. Oshunsanya and OrevaOghene Aliku",authors:[{id:"175778",title:"Dr.",name:"Suarau",middleName:null,surname:"Oshunsanya",slug:"suarau-oshunsanya",fullName:"Suarau Oshunsanya"},{id:"176082",title:"Mr.",name:"OrevaOghene",middleName:null,surname:"Aliku",slug:"orevaoghene-aliku",fullName:"OrevaOghene Aliku"}]}],mostDownloadedChaptersLast30Days:[{id:"62267",title:"Identification of Karst Forms Using LiDAR Technology: Cozumel Island, Mexico",slug:"identification-of-karst-forms-using-lidar-technology-cozumel-island-mexico",totalDownloads:949,totalCrossrefCites:1,totalDimensionsCites:2,abstract:"Morphological relief analysis allows the identification of geomorphological forms and cartographic-environmental studies make extensive use of the medium (1:50,000) and large scale (1:250,000), where the topographical contrast is evident. However, at a detailed scale (<1:20,000) and for territories where the contrast of relief does not exceed 10 m in height, the morphological analyses must be adapted accordingly, because they contribute information to altimetry studies and to the topographic configuration of units. Thus, through visual interpretation and manipulation of high-resolution topographical LiDAR data from Cozumel Island, a relief analysis is presented at a detailed scale for the purpose of recognizing the geomorphological units of karst origin, using altimetry and slope cartography, digital models of elevation, and shading that permits the identification of 109 new exokarstic doline and uvala formations.",book:{id:"7465",slug:"trends-in-geomatics-an-earth-science-perspective",title:"Trends in Geomatics",fullTitle:"Trends in Geomatics - An Earth Science Perspective"},signatures:"Oscar Frausto-Martínez, Norma Angelica Zapi-Salazar and Orlando\nColin-Olivares",authors:[{id:"185429",title:"Dr.",name:"Oscar",middleName:null,surname:"Frausto-Martinez",slug:"oscar-frausto-martinez",fullName:"Oscar Frausto-Martinez"},{id:"258227",title:"MSc.",name:"Orlando",middleName:null,surname:"Colin - Olivares",slug:"orlando-colin-olivares",fullName:"Orlando Colin - Olivares"},{id:"258228",title:"MSc.",name:"Norma Angélica",middleName:null,surname:"Zapi - Salazar",slug:"norma-angelica-zapi-salazar",fullName:"Norma Angélica Zapi - Salazar"}]},{id:"64109",title:"Mathematical Analysis of Some Typical Problems in Geodesy by Means of Computer Algebra",slug:"mathematical-analysis-of-some-typical-problems-in-geodesy-by-means-of-computer-algebra",totalDownloads:922,totalCrossrefCites:0,totalDimensionsCites:1,abstract:"There are many complicated and fussy mathematical analysis processes in geodesy, such as the power series expansions of the ellipsoid’s eccentricity, high order derivation of complex and implicit functions, operation of trigonometric function, expansions of special functions and integral transformation. Taking some typical mathematical analysis processes in geodesy as research objects, the computer algebra analysis are systematically carried out to bread, deep and detailed extent with the help of computer algebra analysis method and the powerful ability of mathematical analysis of computer algebra system. The forward and inverse expansions of the meridian arc in geometric geodesy, the nonsingular expressions of singular integration in physical geodesy and the series expansions of direct transformations between three anomalies in satellite geodesy are established, which have more concise form, stricter theory basis and higher accuracy compared to traditional ones. The breakthrough and innovation of some mathematical analysis problems in the special field of geodesy are realized, which will further enrich and perfect the theoretical system of geodesy.",book:{id:"7465",slug:"trends-in-geomatics-an-earth-science-perspective",title:"Trends in Geomatics",fullTitle:"Trends in Geomatics - An Earth Science Perspective"},signatures:"Hou-pu Li and Shao-feng Bian",authors:[{id:"141289",title:"Prof.",name:"Shao-Feng",middleName:null,surname:"Bian",slug:"shao-feng-bian",fullName:"Shao-Feng Bian"},{id:"141296",title:"Dr.",name:"Hou-Pu",middleName:null,surname:"Li",slug:"hou-pu-li",fullName:"Hou-Pu Li"}]},{id:"51786",title:"GIS Applications in Agronomy",slug:"gis-applications-in-agronomy",totalDownloads:3022,totalCrossrefCites:1,totalDimensionsCites:2,abstract:"Agronomy is a branch of agriculture that deals with soil and crop. Soil varies in space and is responsible for variation in the growth and yield of crops on the field. This variation in the yields of crops planted and monitored on the same parcel of land under the same environmental conditions has been a great concern to farmers. Spatial variations of soil nutrients status, as caused by topography, soil texture and management practices, have been observed across the fields. Hence, the need to separate the field into site specific management units using geographical information systems (GIS) for effective soil and crop management in order to obtain optimum productivity. Over the years, field sizes, farming direction, locations of fences, rotations and fertility programmes have changed the nutritional status of the farms. Consequently, the productivity of the soil has equally been affected. In spite of these factors, conventional agriculture treats an entire field uniformly with respect to the application of fertiliser, pesticides, soil amendments and other chemical application. The use of GIS will help farmers to overcome over- or under-applications of fertiliser and other agrochemical applications. The potential of GIS application in agronomy is obviously large. However, the GIS user community in the field of agronomy is rather small compared to other business sectors. To advance the use of GIS in agronomic studies, this Chapter in book tends to explore the applications of GIS to some fields in agronomy.",book:{id:"5246",slug:"geospatial-technology-environmental-and-social-applications",title:"Geospatial Technology",fullTitle:"Geospatial Technology - Environmental and Social Applications"},signatures:"Suarau O. Oshunsanya and OrevaOghene Aliku",authors:[{id:"175778",title:"Dr.",name:"Suarau",middleName:null,surname:"Oshunsanya",slug:"suarau-oshunsanya",fullName:"Suarau Oshunsanya"},{id:"176082",title:"Mr.",name:"OrevaOghene",middleName:null,surname:"Aliku",slug:"orevaoghene-aliku",fullName:"OrevaOghene Aliku"}]},{id:"51468",title:"Increasing the Adaptive Capacity of Indigenous People to Environmental Change: The Potential Use of an Innovative, Web-Based, Collaborative-Geomatics Informatics Tool to Reduce the Degree of Exposure of First Nations Cree to Hazardous Travel Routes",slug:"increasing-the-adaptive-capacity-of-indigenous-people-to-environmental-change-the-potential-use-of-a",totalDownloads:1433,totalCrossrefCites:0,totalDimensionsCites:0,abstract:"The arctic and subarctic regions of Canada are experiencing amplified climate change impacts, which are disproportionately impacting Canadian indigenous populations’ ability to safely travel on land to acquire resources. Less predictable and more dangerous travel conditions are impacting not only the health and safety of individuals but also the traditional lifestyles that are vital to the cultural well-being of these indigenous communities. The University of Waterloo’s Computer Systems Group has developed a novel decision-support tool termed “Collaborative-Geomatics.” This web-based informatics tool can allow for the community to monitor, in real-time, the safety of travel routes. Using handheld GPS tracking systems, the utility of the geomatics system to present real-time travel conditions was carried out in a Canadian First Nations community, located along the Western James Bay coast. The results of this study showed that the collaborative-geomatics tool offers the potential to monitor and store information on the safety of travel routes, helping to promote adaptive capacity and aid in knowledge transfer within arctic and subarctic indigenous communities.",book:{id:"5246",slug:"geospatial-technology-environmental-and-social-applications",title:"Geospatial Technology",fullTitle:"Geospatial Technology - Environmental and Social Applications"},signatures:"Christine D. Barbeau, Donald Cowan and Leonard J.S. Tsuji",authors:[{id:"181883",title:"Ph.D. Student",name:"Christine",middleName:null,surname:"Barbeau",slug:"christine-barbeau",fullName:"Christine Barbeau"},{id:"186196",title:"Dr.",name:"Don",middleName:null,surname:"Cowan",slug:"don-cowan",fullName:"Don Cowan"}]},{id:"64719",title:"Architectural Design and Prototyping of Co-PPGIS: A Groupware-Based Online Synchronous Collaborative PPGIS to Support Municipality Development and Planning Management Workflows",slug:"architectural-design-and-prototyping-of-co-ppgis-a-groupware-based-online-synchronous-collaborative-",totalDownloads:1064,totalCrossrefCites:1,totalDimensionsCites:1,abstract:"Co-PPGIS has a wide variety of applications like municipal planning, emergency response, public health and security, etc. The main focus of this chapter is on the development and design of a Web Collaborative PPGIS (Co-PPGIS) infrastructure. As part of municipality’s planning and management services, Co-PPGIS is developed for real-time map sharing application system. Co-PPGIS is an effective and essential online meeting system for supporting group collaborations on geographic information such as maps and imageries, and capturing and sharing of local/domain knowledge in real time. Co-PPGIS permits amalgamation of geospatial data and collaborator’s input in the form of geo-referenced notations. It incorporates coherent components as map sharing, real-time chat, video conferencing, geo-referenced textual and graphical notations. The study aims to focus on public participation and geo-collaboration facilitated with information sharing, interactive geo-conferencing, real-time map, and data sharing with tools to draw features or add annotation to the map while discussions, uploading documents, and live communication. Co-PPGIS provides an efficient and reliable platform that will significantly reduce the time to acquire, process, and analyze data. The significance of this study is to contribute to existing public participation practices, to municipal planning, to decision-making, or to geographic information science.",book:{id:"7465",slug:"trends-in-geomatics-an-earth-science-perspective",title:"Trends in Geomatics",fullTitle:"Trends in Geomatics - An Earth Science Perspective"},signatures:"Muhammad A. Butt, Syed Amer Mahmood, Javed Sami, Jahanzeb\nQureshi, Muhammad Kashif Nazir, Amer Masood, Khadija Waheed\nand Aysha Khalid",authors:[{id:"3899",title:"Mr.",name:"Syed Amer",middleName:null,surname:"Mahmood",slug:"syed-amer-mahmood",fullName:"Syed Amer Mahmood"},{id:"242037",title:"Dr.",name:"M. Atif",middleName:null,surname:"Butt",slug:"m.-atif-butt",fullName:"M. Atif Butt"},{id:"269239",title:"Mr.",name:"Javed",middleName:null,surname:"Sami",slug:"javed-sami",fullName:"Javed Sami"},{id:"269240",title:"Mr.",name:"Jahanzeb",middleName:null,surname:"Qureshi",slug:"jahanzeb-qureshi",fullName:"Jahanzeb Qureshi"},{id:"269241",title:"Mr.",name:"Amer",middleName:null,surname:"Masood",slug:"amer-masood",fullName:"Amer Masood"},{id:"270009",title:"Mr.",name:"Muhammad Kashif",middleName:null,surname:"Nazir",slug:"muhammad-kashif-nazir",fullName:"Muhammad Kashif Nazir"}]}],onlineFirstChaptersFilter:{topicId:"785",limit:6,offset:0},onlineFirstChaptersCollection:[],onlineFirstChaptersTotal:0},preDownload:{success:null,errors:{}},subscriptionForm:{success:null,errors:{}},aboutIntechopen:{},privacyPolicy:{},peerReviewing:{},howOpenAccessPublishingWithIntechopenWorks:{},sponsorshipBooks:{sponsorshipBooks:[],offset:8,limit:8,total:0},allSeries:{pteSeriesList:[{id:"14",title:"Artificial Intelligence",numberOfPublishedBooks:8,numberOfPublishedChapters:87,numberOfOpenTopics:6,numberOfUpcomingTopics:0,issn:"2633-1403",doi:"10.5772/intechopen.79920",isOpenForSubmission:!0},{id:"7",title:"Biomedical Engineering",numberOfPublishedBooks:12,numberOfPublishedChapters:98,numberOfOpenTopics:3,numberOfUpcomingTopics:0,issn:"2631-5343",doi:"10.5772/intechopen.71985",isOpenForSubmission:!0}],lsSeriesList:[{id:"11",title:"Biochemistry",numberOfPublishedBooks:27,numberOfPublishedChapters:286,numberOfOpenTopics:4,numberOfUpcomingTopics:0,issn:"2632-0983",doi:"10.5772/intechopen.72877",isOpenForSubmission:!0},{id:"25",title:"Environmental Sciences",numberOfPublishedBooks:1,numberOfPublishedChapters:9,numberOfOpenTopics:4,numberOfUpcomingTopics:0,issn:"2754-6713",doi:"10.5772/intechopen.100362",isOpenForSubmission:!0},{id:"10",title:"Physiology",numberOfPublishedBooks:11,numberOfPublishedChapters:139,numberOfOpenTopics:4,numberOfUpcomingTopics:0,issn:"2631-8261",doi:"10.5772/intechopen.72796",isOpenForSubmission:!0}],hsSeriesList:[{id:"3",title:"Dentistry",numberOfPublishedBooks:8,numberOfPublishedChapters:129,numberOfOpenTopics:0,numberOfUpcomingTopics:2,issn:"2631-6218",doi:"10.5772/intechopen.71199",isOpenForSubmission:!1},{id:"6",title:"Infectious Diseases",numberOfPublishedBooks:13,numberOfPublishedChapters:106,numberOfOpenTopics:3,numberOfUpcomingTopics:1,issn:"2631-6188",doi:"10.5772/intechopen.71852",isOpenForSubmission:!0},{id:"13",title:"Veterinary Medicine and Science",numberOfPublishedBooks:9,numberOfPublishedChapters:101,numberOfOpenTopics:3,numberOfUpcomingTopics:0,issn:"2632-0517",doi:"10.5772/intechopen.73681",isOpenForSubmission:!0}],sshSeriesList:[{id:"22",title:"Business, Management and Economics",numberOfPublishedBooks:1,numberOfPublishedChapters:11,numberOfOpenTopics:2,numberOfUpcomingTopics:1,issn:null,doi:"10.5772/intechopen.100359",isOpenForSubmission:!0},{id:"23",title:"Education and Human Development",numberOfPublishedBooks:0,numberOfPublishedChapters:0,numberOfOpenTopics:2,numberOfUpcomingTopics:0,issn:null,doi:"10.5772/intechopen.100360",isOpenForSubmission:!1},{id:"24",title:"Sustainable Development",numberOfPublishedBooks:0,numberOfPublishedChapters:9,numberOfOpenTopics:4,numberOfUpcomingTopics:1,issn:null,doi:"10.5772/intechopen.100361",isOpenForSubmission:!0}],testimonialsList:[{id:"13",text:"The collaboration with and support of the technical staff of IntechOpen is fantastic. The whole process of submitting an article and editing of the submitted article goes extremely smooth and fast, the number of reads and downloads of chapters is high, and the contributions are also frequently cited.",author:{id:"55578",name:"Antonio",surname:"Jurado-Navas",institutionString:null,profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002bRisIQAS/Profile_Picture_1626166543950",slug:"antonio-jurado-navas",institution:{id:"720",name:"University of Malaga",country:{id:null,name:"Spain"}}}},{id:"6",text:"It is great to work with the IntechOpen to produce a worthwhile collection of research that also becomes a great educational resource and guide for future research endeavors.",author:{id:"259298",name:"Edward",surname:"Narayan",institutionString:null,profilePictureURL:"https://mts.intechopen.com/storage/users/259298/images/system/259298.jpeg",slug:"edward-narayan",institution:{id:"3",name:"University of Queensland",country:{id:null,name:"Australia"}}}}]},series:{item:{id:"11",title:"Biochemistry",doi:"10.5772/intechopen.72877",issn:"2632-0983",scope:"Biochemistry, the study of chemical transformations occurring within living organisms, impacts all areas of life sciences, from molecular crystallography and genetics to ecology, medicine, and population biology. Biochemistry examines macromolecules - proteins, nucleic acids, carbohydrates, and lipids – and their building blocks, structures, functions, and interactions. Much of biochemistry is devoted to enzymes, proteins that catalyze chemical reactions, enzyme structures, mechanisms of action and their roles within cells. Biochemistry also studies small signaling molecules, coenzymes, inhibitors, vitamins, and hormones, which play roles in life processes. Biochemical experimentation, besides coopting classical chemistry methods, e.g., chromatography, adopted new techniques, e.g., X-ray diffraction, electron microscopy, NMR, radioisotopes, and developed sophisticated microbial genetic tools, e.g., auxotroph mutants and their revertants, fermentation, etc. More recently, biochemistry embraced the ‘big data’ omics systems. Initial biochemical studies have been exclusively analytic: dissecting, purifying, and examining individual components of a biological system; in the apt words of Efraim Racker (1913 –1991), “Don’t waste clean thinking on dirty enzymes.” Today, however, biochemistry is becoming more agglomerative and comprehensive, setting out to integrate and describe entirely particular biological systems. The ‘big data’ metabolomics can define the complement of small molecules, e.g., in a soil or biofilm sample; proteomics can distinguish all the comprising proteins, e.g., serum; metagenomics can identify all the genes in a complex environment, e.g., the bovine rumen. This Biochemistry Series will address the current research on biomolecules and the emerging trends with great promise.",coverUrl:"https://cdn.intechopen.com/series/covers/11.jpg",latestPublicationDate:"May 18th, 2022",hasOnlineFirst:!0,numberOfPublishedBooks:27,editor:{id:"31610",title:"Dr.",name:"Miroslav",middleName:null,surname:"Blumenberg",slug:"miroslav-blumenberg",fullName:"Miroslav Blumenberg",profilePictureURL:"https://mts.intechopen.com/storage/users/31610/images/system/31610.jpg",biography:"Miroslav Blumenberg, Ph.D., was born in Subotica and received his BSc in Belgrade, Yugoslavia. He completed his Ph.D. at MIT in Organic Chemistry; he followed up his Ph.D. with two postdoctoral study periods at Stanford University. Since 1983, he has been a faculty member of the RO Perelman Department of Dermatology, NYU School of Medicine, where he is codirector of a training grant in cutaneous biology. Dr. Blumenberg’s research is focused on the epidermis, expression of keratin genes, transcription profiling, keratinocyte differentiation, inflammatory diseases and cancers, and most recently the effects of the microbiome on the skin. He has published more than 100 peer-reviewed research articles and graduated numerous Ph.D. and postdoctoral students.",institutionString:null,institution:{name:"New York University Langone Medical Center",institutionURL:null,country:{name:"United States of America"}}},editorTwo:null,editorThree:null},subseries:{paginationCount:4,paginationItems:[{id:"14",title:"Cell and Molecular Biology",coverUrl:"https://cdn.intechopen.com/series_topics/covers/14.jpg",isOpenForSubmission:!0,editor:{id:"165627",title:"Dr.",name:"Rosa María",middleName:null,surname:"Martínez-Espinosa",slug:"rosa-maria-martinez-espinosa",fullName:"Rosa María Martínez-Espinosa",profilePictureURL:"https://mts.intechopen.com/storage/users/165627/images/system/165627.jpeg",biography:"Dr. Rosa María Martínez-Espinosa has been a Spanish Full Professor since 2020 (Biochemistry and Molecular Biology) and is currently Vice-President of International Relations and Cooperation development and leader of the research group 'Applied Biochemistry” (University of Alicante, Spain). Other positions she has held at the university include Vice-Dean of Master Programs, Vice-Dean of the Degree in Biology and Vice-Dean for Mobility and Enterprise and Engagement at the Faculty of Science (University of Alicante). She received her Bachelor in Biology in 1998 (University of Alicante) and her PhD in 2003 (Biochemistry, University of Alicante). She undertook post-doctoral research at the University of East Anglia (Norwich, U.K. 2004-2005; 2007-2008).\nHer multidisciplinary research focuses on investigating archaea and their potential applications in biotechnology. She has an H-index of 21. She has authored one patent and has published more than 70 indexed papers and around 60 book chapters.\nShe has contributed to more than 150 national and international meetings during the last 15 years. Her research interests include archaea metabolism, enzymes purification and characterization, gene regulation, carotenoids and bioplastics production, antioxidant\ncompounds, waste water treatments, and brines bioremediation.\nRosa María’s other roles include editorial board member for several journals related\nto biochemistry, reviewer for more than 60 journals (biochemistry, molecular biology, biotechnology, chemistry and microbiology) and president of several organizing committees in international meetings related to the N-cycle or respiratory processes.",institutionString:null,institution:{name:"University of Alicante",institutionURL:null,country:{name:"Spain"}}},editorTwo:null,editorThree:null},{id:"15",title:"Chemical Biology",coverUrl:"https://cdn.intechopen.com/series_topics/covers/15.jpg",isOpenForSubmission:!0,editor:{id:"441442",title:"Dr.",name:"Şükrü",middleName:null,surname:"Beydemir",slug:"sukru-beydemir",fullName:"Şükrü Beydemir",profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0033Y00003GsUoIQAV/Profile_Picture_1634557147521",biography:"Dr. Şükrü Beydemir obtained a BSc in Chemistry in 1995 from Yüzüncü Yıl University, MSc in Biochemistry in 1998, and PhD in Biochemistry in 2002 from Atatürk University, Turkey. He performed post-doctoral studies at Max-Planck Institute, Germany, and University of Florence, Italy in addition to making several scientific visits abroad. He currently works as a Full Professor of Biochemistry in the Faculty of Pharmacy, Anadolu University, Turkey. Dr. Beydemir has published over a hundred scientific papers spanning protein biochemistry, enzymology and medicinal chemistry, reviews, book chapters and presented several conferences to scientists worldwide. He has received numerous publication awards from various international scientific councils. He serves in the Editorial Board of several international journals. Dr. Beydemir is also Rector of Bilecik Şeyh Edebali University, Turkey.",institutionString:null,institution:{name:"Anadolu University",institutionURL:null,country:{name:"Turkey"}}},editorTwo:{id:"13652",title:"Prof.",name:"Deniz",middleName:null,surname:"Ekinci",slug:"deniz-ekinci",fullName:"Deniz Ekinci",profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002aYLT1QAO/Profile_Picture_1634557223079",biography:"Dr. Deniz Ekinci obtained a BSc in Chemistry in 2004, MSc in Biochemistry in 2006, and PhD in Biochemistry in 2009 from Atatürk University, Turkey. He studied at Stetson University, USA, in 2007-2008 and at the Max Planck Institute of Molecular Cell Biology and Genetics, Germany, in 2009-2010. Dr. Ekinci currently works as a Full Professor of Biochemistry in the Faculty of Agriculture and is the Head of the Enzyme and Microbial Biotechnology Division, Ondokuz Mayıs University, Turkey. He is a member of the Turkish Biochemical Society, American Chemical Society, and German Genetics society. Dr. Ekinci published around ninety scientific papers, reviews and book chapters, and presented several conferences to scientists. He has received numerous publication awards from several scientific councils. Dr. Ekinci serves as the Editor in Chief of four international books and is involved in the Editorial Board of several international journals.",institutionString:null,institution:{name:"Ondokuz Mayıs University",institutionURL:null,country:{name:"Turkey"}}},editorThree:null},{id:"17",title:"Metabolism",coverUrl:"https://cdn.intechopen.com/series_topics/covers/17.jpg",isOpenForSubmission:!0,editor:{id:"138626",title:"Dr.",name:"Yannis",middleName:null,surname:"Karamanos",slug:"yannis-karamanos",fullName:"Yannis Karamanos",profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002g6Jv2QAE/Profile_Picture_1629356660984",biography:"Yannis Karamanos, born in Greece in 1953, completed his pre-graduate studies at the Université Pierre et Marie Curie, Paris, then his Masters and Doctoral degree at the Université de Lille (1983). He was associate professor at the University of Limoges (1987) before becoming full professor of biochemistry at the Université d’Artois (1996). He worked on the structure-function relationships of glycoconjugates and his main project was the investigations on the biological roles of the de-N-glycosylation enzymes (Endo-N-acetyl-β-D-glucosaminidase and peptide-N4-(N-acetyl-β-glucosaminyl) asparagine amidase). From 2002 he contributes to the understanding of the Blood-brain barrier functioning using proteomics approaches. He has published more than 70 papers. His teaching areas are energy metabolism and regulation, integration and organ specialization and metabolic adaptation.",institutionString:null,institution:{name:"Artois University",institutionURL:null,country:{name:"France"}}},editorTwo:null,editorThree:null},{id:"18",title:"Proteomics",coverUrl:"https://cdn.intechopen.com/series_topics/covers/18.jpg",isOpenForSubmission:!0,editor:{id:"200689",title:"Prof.",name:"Paolo",middleName:null,surname:"Iadarola",slug:"paolo-iadarola",fullName:"Paolo Iadarola",profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002bSCl8QAG/Profile_Picture_1623568118342",biography:"Paolo Iadarola graduated with a degree in Chemistry from the University of Pavia (Italy) in July 1972. He then worked as an Assistant Professor at the Faculty of Science of the same University until 1984. In 1985, Prof. Iadarola became Associate Professor at the Department of Biology and Biotechnologies of the University of Pavia and retired in October 2017. Since then, he has been working as an Adjunct Professor in the same Department at the University of Pavia. His research activity during the first years was primarily focused on the purification and structural characterization of enzymes from animal and plant sources. During this period, Prof. Iadarola familiarized himself with the conventional techniques used in column chromatography, spectrophotometry, manual Edman degradation, and electrophoresis). Since 1995, he has been working on: i) the determination in biological fluids (serum, urine, bronchoalveolar lavage, sputum) of proteolytic activities involved in the degradation processes of connective tissue matrix, and ii) on the identification of biological markers of lung diseases. In this context, he has developed and validated new methodologies (e.g., Capillary Electrophoresis coupled to Laser-Induced Fluorescence, CE-LIF) whose application enabled him to determine both the amounts of biochemical markers (Desmosines) in urine/serum of patients affected by Chronic Obstructive Pulmonary Disease (COPD) and the activity of proteolytic enzymes (Human Neutrophil Elastase, Cathepsin G, Pseudomonas aeruginosa elastase) in sputa of these patients. More recently, Prof. Iadarola was involved in developing techniques such as two-dimensional electrophoresis coupled to liquid chromatography/mass spectrometry (2DE-LC/MS) for the proteomic analysis of biological fluids aimed at the identification of potential biomarkers of different lung diseases. He is the author of about 150 publications (According to Scopus: H-Index: 23; Total citations: 1568- According to WOS: H-Index: 20; Total Citations: 1296) of peer-reviewed international journals. He is a Consultant Reviewer for several journals, including the Journal of Chromatography A, Journal of Chromatography B, Plos ONE, Proteomes, International Journal of Molecular Science, Biotech, Electrophoresis, and others. He is also Associate Editor of Biotech.",institutionString:null,institution:{name:"University of Pavia",institutionURL:null,country:{name:"Italy"}}},editorTwo:{id:"201414",title:"Dr.",name:"Simona",middleName:null,surname:"Viglio",slug:"simona-viglio",fullName:"Simona Viglio",profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002bRKDHQA4/Profile_Picture_1630402531487",biography:"Simona Viglio is an Associate Professor of Biochemistry at the Department of Molecular Medicine at the University of Pavia. She has been working since 1995 on the determination of proteolytic enzymes involved in the degradation process of connective tissue matrix and on the identification of biological markers of lung diseases. She gained considerable experience in developing and validating new methodologies whose applications allowed her to determine both the amount of biomarkers (Desmosine and Isodesmosine) in the urine of patients affected by COPD, and the activity of proteolytic enzymes (HNE, Cathepsin G, Pseudomonas aeruginosa elastase) in the sputa of these patients. Simona Viglio was also involved in research dealing with the supplementation of amino acids in patients with brain injury and chronic heart failure. She is presently engaged in the development of 2-DE and LC-MS techniques for the study of proteomics in biological fluids. The aim of this research is the identification of potential biomarkers of lung diseases. She is an author of about 90 publications (According to Scopus: H-Index: 23; According to WOS: H-Index: 20) on peer-reviewed journals, a member of the “Società Italiana di Biochimica e Biologia Molecolare,“ and a Consultant Reviewer for International Journal of Molecular Science, Journal of Chromatography A, COPD, Plos ONE and Nutritional Neuroscience.",institutionString:null,institution:{name:"University of Pavia",institutionURL:null,country:{name:"Italy"}}},editorThree:null}]},overviewPageOFChapters:{paginationCount:49,paginationItems:[{id:"80495",title:"Iron in Cell Metabolism and Disease",doi:"10.5772/intechopen.101908",signatures:"Eeka Prabhakar",slug:"iron-in-cell-metabolism-and-disease",totalDownloads:0,totalCrossrefCites:null,totalDimensionsCites:null,authors:null,book:{title:"Iron Metabolism - Iron a Double‐Edged Sword",coverURL:"https://cdn.intechopen.com/books/images_new/10842.jpg",subseries:{id:"17",title:"Metabolism"}}},{id:"81799",title:"Cross Talk of Purinergic and Immune Signaling: Implication in Inflammatory and Pathogenic Diseases",doi:"10.5772/intechopen.104978",signatures:"Richa Rai",slug:"cross-talk-of-purinergic-and-immune-signaling-implication-in-inflammatory-and-pathogenic-diseases",totalDownloads:7,totalCrossrefCites:0,totalDimensionsCites:0,authors:null,book:{title:"Purinergic System",coverURL:"https://cdn.intechopen.com/books/images_new/10801.jpg",subseries:{id:"17",title:"Metabolism"}}},{id:"81764",title:"Involvement of the Purinergic System in Cell Death in Models of Retinopathies",doi:"10.5772/intechopen.103935",signatures:"Douglas Penaforte Cruz, Marinna Garcia Repossi and Lucianne Fragel Madeira",slug:"involvement-of-the-purinergic-system-in-cell-death-in-models-of-retinopathies",totalDownloads:4,totalCrossrefCites:0,totalDimensionsCites:0,authors:null,book:{title:"Purinergic System",coverURL:"https://cdn.intechopen.com/books/images_new/10801.jpg",subseries:{id:"17",title:"Metabolism"}}},{id:"81756",title:"Alteration of Cytokines Level and Oxidative Stress Parameters in COVID-19",doi:"10.5772/intechopen.104950",signatures:"Marija Petrusevska, Emilija Atanasovska, Dragica Zendelovska, Aleksandar Eftimov and Katerina Spasovska",slug:"alteration-of-cytokines-level-and-oxidative-stress-parameters-in-covid-19",totalDownloads:8,totalCrossrefCites:0,totalDimensionsCites:0,authors:null,book:{title:"Chemokines Updates",coverURL:"https://cdn.intechopen.com/books/images_new/11672.jpg",subseries:{id:"18",title:"Proteomics"}}}]},overviewPagePublishedBooks:{paginationCount:27,paginationItems:[{type:"book",id:"7006",title:"Biochemistry and Health Benefits of Fatty Acids",subtitle:null,coverURL:"https://cdn.intechopen.com/books/images_new/7006.jpg",slug:"biochemistry-and-health-benefits-of-fatty-acids",publishedDate:"December 19th 2018",editedByType:"Edited by",bookSignature:"Viduranga Waisundara",hash:"c93a00abd68b5eba67e5e719f67fd20b",volumeInSeries:1,fullTitle:"Biochemistry and Health Benefits of Fatty Acids",editors:[{id:"194281",title:"Dr.",name:"Viduranga Y.",middleName:null,surname:"Waisundara",slug:"viduranga-y.-waisundara",fullName:"Viduranga Y. Waisundara",profilePictureURL:"https://mts.intechopen.com/storage/users/194281/images/system/194281.jpg",biography:"Dr. Viduranga Waisundara obtained her Ph.D. in Food Science and Technology from the Department of Chemistry, National University of Singapore, in 2010. She was a lecturer at Temasek Polytechnic, Singapore from July 2009 to March 2013. She relocated to her motherland of Sri Lanka and spearheaded the Functional Food Product Development Project at the National Institute of Fundamental Studies from April 2013 to October 2016. She was a senior lecturer on a temporary basis at the Department of Food Technology, Faculty of Technology, Rajarata University of Sri Lanka. She is currently Deputy Principal of the Australian College of Business and Technology – Kandy Campus, Sri Lanka. She is also the Global Harmonization Initiative (GHI) Ambassador to Sri Lanka.",institutionString:"Australian College of Business & Technology",institution:null}]},{type:"book",id:"6820",title:"Keratin",subtitle:null,coverURL:"https://cdn.intechopen.com/books/images_new/6820.jpg",slug:"keratin",publishedDate:"December 19th 2018",editedByType:"Edited by",bookSignature:"Miroslav Blumenberg",hash:"6def75cd4b6b5324a02b6dc0359896d0",volumeInSeries:2,fullTitle:"Keratin",editors:[{id:"31610",title:"Dr.",name:"Miroslav",middleName:null,surname:"Blumenberg",slug:"miroslav-blumenberg",fullName:"Miroslav Blumenberg",profilePictureURL:"https://mts.intechopen.com/storage/users/31610/images/system/31610.jpg",biography:"Miroslav Blumenberg, Ph.D., was born in Subotica and received his BSc in Belgrade, Yugoslavia. He completed his Ph.D. at MIT in Organic Chemistry; he followed up his Ph.D. with two postdoctoral study periods at Stanford University. Since 1983, he has been a faculty member of the RO Perelman Department of Dermatology, NYU School of Medicine, where he is codirector of a training grant in cutaneous biology. Dr. Blumenberg’s research is focused on the epidermis, expression of keratin genes, transcription profiling, keratinocyte differentiation, inflammatory diseases and cancers, and most recently the effects of the microbiome on the skin. He has published more than 100 peer-reviewed research articles and graduated numerous Ph.D. and postdoctoral students.",institutionString:null,institution:{name:"New York University Langone Medical Center",institutionURL:null,country:{name:"United States of America"}}}]},{type:"book",id:"7978",title:"Vitamin A",subtitle:null,coverURL:"https://cdn.intechopen.com/books/images_new/7978.jpg",slug:"vitamin-a",publishedDate:"May 15th 2019",editedByType:"Edited by",bookSignature:"Leila Queiroz Zepka, Veridiana Vera de Rosso and Eduardo Jacob-Lopes",hash:"dad04a658ab9e3d851d23705980a688b",volumeInSeries:3,fullTitle:"Vitamin A",editors:[{id:"261969",title:"Dr.",name:"Leila",middleName:null,surname:"Queiroz Zepka",slug:"leila-queiroz-zepka",fullName:"Leila Queiroz Zepka",profilePictureURL:"https://mts.intechopen.com/storage/users/261969/images/system/261969.png",biography:"Prof. Dr. Leila Queiroz Zepka is currently an associate professor in the Department of Food Technology and Science, Federal University of Santa Maria, Brazil. She has more than fifteen years of teaching and research experience. She has published more than 550 scientific publications/communications, including 15 books, 50 book chapters, 100 original research papers, 380 research communications in national and international conferences, and 12 patents. She is a member of the editorial board of five journals and acts as a reviewer for several national and international journals. Her research interests include microalgal biotechnology with an emphasis on microalgae-based products.",institutionString:"Universidade Federal de Santa Maria",institution:{name:"Universidade Federal de Santa Maria",institutionURL:null,country:{name:"Brazil"}}}]},{type:"book",id:"7953",title:"Bioluminescence",subtitle:"Analytical Applications and Basic Biology",coverURL:"https://cdn.intechopen.com/books/images_new/7953.jpg",slug:"bioluminescence-analytical-applications-and-basic-biology",publishedDate:"September 25th 2019",editedByType:"Edited by",bookSignature:"Hirobumi Suzuki",hash:"3a8efa00b71abea11bf01973dc589979",volumeInSeries:4,fullTitle:"Bioluminescence - Analytical Applications and Basic Biology",editors:[{id:"185746",title:"Dr.",name:"Hirobumi",middleName:null,surname:"Suzuki",slug:"hirobumi-suzuki",fullName:"Hirobumi Suzuki",profilePictureURL:"https://mts.intechopen.com/storage/users/185746/images/system/185746.png",biography:"Dr. Hirobumi Suzuki received his Ph.D. in 1997 from Tokyo Metropolitan University, Japan, where he studied firefly phylogeny and the evolution of mating systems. He is especially interested in the genetic differentiation pattern and speciation process that correlate to the flashing pattern and mating behavior of some fireflies in Japan. He then worked for Olympus Corporation, a Japanese manufacturer of optics and imaging products, where he was involved in the development of luminescence technology and produced a bioluminescence microscope that is currently being used for gene expression analysis in chronobiology, neurobiology, and developmental biology. Dr. Suzuki currently serves as a visiting researcher at Kogakuin University, Japan, and also a vice president of the Japan Firefly Society.",institutionString:"Kogakuin University",institution:null}]}]},openForSubmissionBooks:{paginationCount:6,paginationItems:[{id:"11667",title:"Marine Pollution - Recent Developments",coverURL:"https://cdn.intechopen.com/books/images_new/11667.jpg",hash:"e524cd97843b075a724e151256773631",secondStepPassed:!0,currentStepOfPublishingProcess:3,submissionDeadline:"April 20th 2022",isOpenForSubmission:!0,editors:[{id:"318562",title:"Dr.",name:"Monique",surname:"Mancuso",slug:"monique-mancuso",fullName:"Monique Mancuso"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null},{id:"11664",title:"Recent Advances in Sensing Technologies for Environmental Control and Monitoring",coverURL:"https://cdn.intechopen.com/books/images_new/11664.jpg",hash:"cf1ee76443e393bc7597723c3ee3e26f",secondStepPassed:!0,currentStepOfPublishingProcess:3,submissionDeadline:"May 4th 2022",isOpenForSubmission:!0,editors:[{id:"24687",title:"Dr.",name:"Toonika",surname:"Rinken",slug:"toonika-rinken",fullName:"Toonika Rinken"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null},{id:"11662",title:"Limnology - The Importance of Monitoring and Correlations of Lentic and Lotic Waters",coverURL:"https://cdn.intechopen.com/books/images_new/11662.jpg",hash:"f1043cf6b1daae7a7b527e1d162ca4a8",secondStepPassed:!0,currentStepOfPublishingProcess:3,submissionDeadline:"May 10th 2022",isOpenForSubmission:!0,editors:[{id:"315689",title:"Dr.",name:"Carmine",surname:"Massarelli",slug:"carmine-massarelli",fullName:"Carmine Massarelli"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null},{id:"10845",title:"Marine Ecosystems - Biodiversity, Ecosystem Services and Human Impacts",coverURL:"https://cdn.intechopen.com/books/images_new/10845.jpg",hash:"727e7eb3d4ba529ec5eb4f150e078523",secondStepPassed:!0,currentStepOfPublishingProcess:3,submissionDeadline:"May 12th 2022",isOpenForSubmission:!0,editors:[{id:"320124",title:"Dr.",name:"Ana M.M.",surname:"Gonçalves",slug:"ana-m.m.-goncalves",fullName:"Ana M.M. Gonçalves"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null},{id:"11665",title:"Recent Advances in Wildlife Management",coverURL:"https://cdn.intechopen.com/books/images_new/11665.jpg",hash:"73da0df494a1a56ab9c4faf2ee811899",secondStepPassed:!1,currentStepOfPublishingProcess:2,submissionDeadline:"May 25th 2022",isOpenForSubmission:!0,editors:[{id:"75563",title:"Dr.",name:"Farzana Khan",surname:"Perveen",slug:"farzana-khan-perveen",fullName:"Farzana Khan Perveen"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null},{id:"11666",title:"Soil Contamination - Recent Advances and Future Perspectives",coverURL:"https://cdn.intechopen.com/books/images_new/11666.jpg",hash:"c8890038b86fb6e5af16ea3c22669ae9",secondStepPassed:!1,currentStepOfPublishingProcess:2,submissionDeadline:"June 9th 2022",isOpenForSubmission:!0,editors:[{id:"299110",title:"Dr.",name:"Adnan",surname:"Mustafa",slug:"adnan-mustafa",fullName:"Adnan Mustafa"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null}]},onlineFirstChapters:{paginationCount:43,paginationItems:[{id:"81796",title:"Apoptosis-Related Diseases and Peroxisomes",doi:"10.5772/intechopen.105052",signatures:"Meimei Wang, Yakun Liu, Ni Chen, Juan Wang and Ye Zhao",slug:"apoptosis-related-diseases-and-peroxisomes",totalDownloads:2,totalCrossrefCites:0,totalDimensionsCites:0,authors:null,book:{title:"The Metabolic Role of Peroxisome in Health and Disease",coverURL:"https://cdn.intechopen.com/books/images_new/10837.jpg",subseries:{id:"11",title:"Cell Physiology"}}},{id:"81723",title:"Peroxisomal Modulation as Therapeutic Alternative for Tackling Multiple Cancers",doi:"10.5772/intechopen.104873",signatures:"Shazia Usmani, Shadma Wahab, Abdul Hafeez, Shabana Khatoon and Syed Misbahul Hasan",slug:"peroxisomal-modulation-as-therapeutic-alternative-for-tackling-multiple-cancers",totalDownloads:3,totalCrossrefCites:0,totalDimensionsCites:0,authors:null,book:{title:"The Metabolic Role of Peroxisome in Health and Disease",coverURL:"https://cdn.intechopen.com/books/images_new/10837.jpg",subseries:{id:"11",title:"Cell Physiology"}}},{id:"81638",title:"Aging and Neuropsychiatric Disease: A General Overview of Prevalence and Trends",doi:"10.5772/intechopen.103102",signatures:"Jelena Milić",slug:"aging-and-neuropsychiatric-disease-a-general-overview-of-prevalence-and-trends",totalDownloads:14,totalCrossrefCites:0,totalDimensionsCites:0,authors:null,book:{title:"Senescence",coverURL:"https://cdn.intechopen.com/books/images_new/10935.jpg",subseries:{id:"11",title:"Cell Physiology"}}},{id:"81566",title:"New and Emerging Technologies for Integrative Ambulatory Autonomic Assessment and Intervention as a Catalyst in the Synergy of Remote Geocoded Biosensing, Algorithmic Networked Cloud Computing, Deep Learning, and Regenerative/Biomic Medicine: Further Real",doi:"10.5772/intechopen.104092",signatures:"Robert L. Drury",slug:"new-and-emerging-technologies-for-integrative-ambulatory-autonomic-assessment-and-intervention-as-a-",totalDownloads:9,totalCrossrefCites:0,totalDimensionsCites:0,authors:null,book:{title:"Autonomic Nervous System - Special Interest Topics",coverURL:"https://cdn.intechopen.com/books/images_new/10835.jpg",subseries:{id:"12",title:"Human Physiology"}}},{id:"81576",title:"Carotenoids in Thermal Adaptation of Plants and Animals",doi:"10.5772/intechopen.104537",signatures:"Ivan M. Petyaev",slug:"carotenoids-in-thermal-adaptation-of-plants-and-animals",totalDownloads:15,totalCrossrefCites:0,totalDimensionsCites:0,authors:[{name:"Ivan",surname:"Petyaev"}],book:{title:"Carotenoids - New Perspectives and Application",coverURL:"https://cdn.intechopen.com/books/images_new/10836.jpg",subseries:{id:"13",title:"Plant Physiology"}}},{id:"81358",title:"New Insights on Carotenoid Production by Gordonia alkanivorans Strain 1B",doi:"10.5772/intechopen.103919",signatures:"Tiago P. Silva, Susana M. Paixão, Ana S. Fernandes, José C. Roseiro and Luís Alves",slug:"new-insights-on-carotenoid-production-by-gordonia-alkanivorans-strain-1b",totalDownloads:13,totalCrossrefCites:0,totalDimensionsCites:0,authors:null,book:{title:"Carotenoids - New Perspectives and Application",coverURL:"https://cdn.intechopen.com/books/images_new/10836.jpg",subseries:{id:"13",title:"Plant Physiology"}}},{id:"81298",title:"Roles of Extracellular Vesicles in Cancer Metastasis",doi:"10.5772/intechopen.103798",signatures:"Eman Helmy Thabet",slug:"roles-of-extracellular-vesicles-in-cancer-metastasis",totalDownloads:20,totalCrossrefCites:0,totalDimensionsCites:0,authors:null,book:{title:"Extracellular Vesicles - Role in Diseases Pathogenesis and Therapy",coverURL:"https://cdn.intechopen.com/books/images_new/10796.jpg",subseries:{id:"11",title:"Cell Physiology"}}},{id:"81290",title:"Musculoskeletal Abnormalities Caused by Cystic Fibrosis",doi:"10.5772/intechopen.104591",signatures:"Mark Lambrechts",slug:"musculoskeletal-abnormalities-caused-by-cystic-fibrosis",totalDownloads:12,totalCrossrefCites:0,totalDimensionsCites:0,authors:null,book:{title:"Advances in Skeletal Muscle Health and Disease",coverURL:"https://cdn.intechopen.com/books/images_new/11675.jpg",subseries:{id:"11",title:"Cell Physiology"}}},{id:"81286",title:"Potassium Derangements: A Pathophysiological Review, Diagnostic Approach, and Clinical Management",doi:"10.5772/intechopen.103016",signatures:"Sairah Sharif and Jie Tang",slug:"potassium-derangements-a-pathophysiological-review-diagnostic-approach-and-clinical-management",totalDownloads:24,totalCrossrefCites:0,totalDimensionsCites:0,authors:null,book:{title:"Potassium in Human Health",coverURL:"https://cdn.intechopen.com/books/images_new/10794.jpg",subseries:{id:"12",title:"Human Physiology"}}},{id:"81246",title:"Role of Carotenoids in Cardiovascular Disease",doi:"10.5772/intechopen.102750",signatures:"Arslan Ahmad, Sakhawat Riaz, Muhammad Shahzaib Nadeem, Umber Mubeen and Khadija Maham",slug:"role-of-carotenoids-in-cardiovascular-disease",totalDownloads:26,totalCrossrefCites:0,totalDimensionsCites:0,authors:null,book:{title:"Carotenoids - New Perspectives and Application",coverURL:"https://cdn.intechopen.com/books/images_new/10836.jpg",subseries:{id:"13",title:"Plant Physiology"}}}]},subseriesFiltersForOFChapters:[{caption:"Plant Physiology",value:13,count:5,group:"subseries"},{caption:"Human Physiology",value:12,count:13,group:"subseries"},{caption:"Cell Physiology",value:11,count:25,group:"subseries"}],publishedBooks:{paginationCount:0,paginationItems:[]},subseriesFiltersForPublishedBooks:[],publicationYearFilters:[],authors:{paginationCount:25,paginationItems:[{id:"429683",title:"Dr.",name:"Bilal",middleName:null,surname:"Khalid",slug:"bilal-khalid",fullName:"Bilal Khalid",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/429683/images/system/429683.png",biography:"Dr. Bilal Khalid received a Ph.D. in Industrial Business Administration from KMITL Business School, Bangkok, in 2021, and a master’s in International Business Management from Stamford International University, Bangkok, in 2017. Dr. Khalid\\'s research interests include leadership and negotiations, digital transformations, gamification, eLearning, blockchain, Big Data, and management of information technology. Dr. Bilal Khalid also serves as an academic editor at Education Research International and a reviewer for international journals.",institutionString:"KMITL Business School",institution:{name:"King Mongkut's Institute of Technology Ladkrabang",country:{name:"Thailand"}}},{id:"418514",title:"Dr.",name:"Muhammad",middleName:null,surname:"Mohiuddin",slug:"muhammad-mohiuddin",fullName:"Muhammad Mohiuddin",position:null,profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0033Y000038UqSfQAK/Profile_Picture_2022-05-13T10:39:03.jpg",biography:"Dr. Muhammad Mohiuddin is an Associate Professor of International Business at Laval University, Canada. He has taught at Thompson Rivers University, Canada; University of Paris-Est, France; Osnabruck University of Applied Science, Germany; and Shanghai Institute of Technology and Tianjin University of Technology, China. He has published research in Research Policy, Applied Economics, Review of Economic Philosophy, Strategic Change, International Journal of Logistics, Sustainability, Journal of Environmental Management, Journal of Global Information Management, Journal of Cleaner Production, M@N@GEMENT, and more. He is a member of CEDIMES Institut (France), Academy of International Business (AIB), Strategic Management Society (SMS), Academy of Management (AOM), Administrative Science Association of Canada (ASAC), and Canadian council of small business and entrepreneurship (CCSBE). He is currently the director of the Research Group on Contemporary Asia (GERAC) at Laval University. He is also co-managing editor of Transnational Corporations Review and a guest editor for Electronic Commerce Research and Journal of Internet Technology.",institutionString:null,institution:{name:"Université Laval",country:{name:"Canada"}}},{id:"189147",title:"Dr.",name:"Hailan",middleName:null,surname:"Salamun",slug:"hailan-salamun",fullName:"Hailan Salamun",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/189147/images/19274_n.jpeg",biography:"Hailan Salamun, (Dr.) was born in Selangor, Malaysia and graduated from Tunku Ampuan Jamaah Religious High School at Shah Alam. Obtained a degree from the International Islamic University (UIA), Gombak in the field of Islamic Revealed Knowledge and Heritage. Next, I furthered my studies to the professional level to obtain a Diploma in Education at UIA. After serving for several years in school, I furthered my studies to the Master of Dakwah and Leadership at Universiti Kebangsaan Malaysia (UKM), Bangi. I graduated with a Doctor of Philosophy in Principalship Leadership from the University of Malaya (UM) in 2010. I am currently a senior lecturer in the Department of Nationalism and Civilization, Center for Basic and Continuing Education, Universiti Malaysia Terengganu. Prior to that, I had served in several educational institutions such as schools, the Institute of Teacher Education (IPG), and also the University of Malaya. I am also actively involved in paper presentation, writing and publishing. My research interests are focused on leadership, education, society and Islamic civilization. This area of research requires a detailed understanding of Islamic studies and research studies in leadership. Another research interest that I have explored recently is the politics of the Malay community and also the leadership of the mosque.",institutionString:null,institution:{name:"Universiti Malaysia Terengganu",country:{name:"Malaysia"}}},{id:"442081",title:"Dr.",name:"Audrey",middleName:null,surname:"Addy",slug:"audrey-addy",fullName:"Audrey Addy",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Kwame Nkrumah University of Science and Technology",country:{name:"Ghana"}}},{id:"437993",title:"Mr.",name:"Job",middleName:null,surname:"Jackson",slug:"job-jackson",fullName:"Job Jackson",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Management College of Southern Africa",country:{name:"South Africa"}}},{id:"428495",title:"Prof.",name:"Asyraf",middleName:null,surname:"Ab Rahman",slug:"asyraf-ab-rahman",fullName:"Asyraf Ab Rahman",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Universiti Malaysia Terengganu",country:{name:"Malaysia"}}},{id:"429650",title:"Dr.",name:"Jacqueline",middleName:null,surname:"Kareem",slug:"jacqueline-kareem",fullName:"Jacqueline Kareem",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Christ University",country:{name:"India"}}},{id:"421041",title:"Dr.",name:"Sunil",middleName:null,surname:"Kumar Ramdas",slug:"sunil-kumar-ramdas",fullName:"Sunil Kumar Ramdas",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Jain University",country:{name:"India"}}},{id:"421833",title:"Mr.",name:"Eugene",middleName:null,surname:"Owusu-Acheampong",slug:"eugene-owusu-acheampong",fullName:"Eugene Owusu-Acheampong",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"University of Ghana",country:{name:"Ghana"}}},{id:"239876",title:"Ph.D.",name:"Luciana",middleName:null,surname:"Mourão",slug:"luciana-mourao",fullName:"Luciana Mourão",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Universidade Salgado de Oliveira",country:{name:"Brazil"}}},{id:"421735",title:"Dr.",name:"elizabeth",middleName:null,surname:"addy",slug:"elizabeth-addy",fullName:"elizabeth addy",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:null},{id:"442083",title:"Dr.",name:"James",middleName:null,surname:"Addy",slug:"james-addy",fullName:"James Addy",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:null},{id:"437991",title:"Prof.",name:"Muhammad",middleName:null,surname:"Hoque",slug:"muhammad-hoque",fullName:"Muhammad Hoque",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:null},{id:"421006",title:"Dr.",name:"Anna",middleName:null,surname:"Uster",slug:"anna-uster",fullName:"Anna Uster",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:null},{id:"470243",title:"Dr.",name:"Md Samim",middleName:null,surname:"Al Azad",slug:"md-samim-al-azad",fullName:"Md Samim Al Azad",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:null},{id:"470244",title:"Dr.",name:"Slimane",middleName:null,surname:"Ed-dafali",slug:"slimane-ed-dafali",fullName:"Slimane Ed-dafali",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:null},{id:"421011",title:"Dr.",name:"Afatakpa",middleName:null,surname:"Fortune",slug:"afatakpa-fortune",fullName:"Afatakpa Fortune",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:null},{id:"446057",title:"Mr.",name:"Okedare",middleName:null,surname:"David Olubukunmi",slug:"okedare-david-olubukunmi",fullName:"Okedare David Olubukunmi",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:null},{id:"421778",title:"Dr.",name:"Fatimah",middleName:"Saeed",surname:"AlAhmari",slug:"fatimah-alahmari",fullName:"Fatimah AlAhmari",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:null},{id:"421024",title:"Prof.",name:"Harold Andrew",middleName:null,surname:"Patrick",slug:"harold-andrew-patrick",fullName:"Harold Andrew Patrick",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:null},{id:"421065",title:"Ms.",name:"Euzália",middleName:null,surname:"do Rosário Botelho Tomé",slug:"euzalia-do-rosario-botelho-tome",fullName:"Euzália do Rosário Botelho Tomé",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:null},{id:"421053",title:"Associate Prof.",name:"Ken",middleName:null,surname:"Kalala Ndalamba",slug:"ken-kalala-ndalamba",fullName:"Ken Kalala Ndalamba",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:null},{id:"421826",title:"Dr.",name:"Inusah",middleName:null,surname:"Salifu",slug:"inusah-salifu",fullName:"Inusah Salifu",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:null},{id:"420823",title:"Prof.",name:"Gardênia da Silva",middleName:null,surname:"Abbad",slug:"gardenia-da-silva-abbad",fullName:"Gardênia da Silva Abbad",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:null},{id:"437613",title:"MSc.",name:"Juliana",middleName:null,surname:"Legentil",slug:"juliana-legentil",fullName:"Juliana Legentil",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:null}]}},subseries:{item:{id:"86",type:"subseries",title:"Business and Management",keywords:"Demographic shifts, Innovation, Technology, Next-gen leaders, Worldwide environmental issues and clean technology, Uncertainty and political risks, Radical adjacency, Emergence of new business ecosystem type, Emergence of different leader and leader values types, Universal connector, Elastic enterprise, Business platform, Supply chain complexity",scope:"