Virulence factors of uropathogenic
\r\n\tMore than 20 species of Leishmania cause different forms of leishmaniasis that range in severity from skin lesions to systemic infections. The variety of leishmaniasis clinical manifestations are related to the parasite species and to the host characteristics, such as genetic background, nutritional status, as well as immunocompetence, and environmental and social factors of the host.
\r\n\tThe current treatment is still based on pentavalent antimonials, which have been in use for more than 100 years, and like the other drugs used in leishmaniasis treatment, such as amphotericin B, pentamidine, and miltefosine, induce several adverse drug effects.
\r\n\tThese characteristics are associated with the increasing number of cases of resistance to current treatments, immunosuppressed patients, and those with hepatic and renal disorders, showing the necessity for research into new therapy options that are more efficient and non-toxic.
\r\n\tThe book is intended to collect chapters on interdisciplinary approaches covering host-parasite and vector interaction, the immune response of hosts, new molecular pathways for parasite survival and persistence, new drug development, and mechanisms of antileishmanial drugs, as well as control, epidemiology, and vaccine development.
Almost four decades after the initial success [1], production of genetically modified plants still takes a central place in the experimental studies and biotechnology of plants. Genetic engineering has made possible introducing beneficial traits from unrelated plants, bacteria, viruses, fungi, or animal species, to overcome the major limitations of conventional plant breeding. Introduction of one or more genes into commercial crop species has helped boost crop yields due to increased resistance of transgenic lines to abiotic stress, pests and pathogens, and manipulation of metabolic pathways resulted in improving the nutritional or industrial value of genetically modified plants. Also, plant “factories” have been designed to produce high amounts of various pharmacologically important compounds, nutrients or other useful substances.
Genetically modified (GM) crops have been cultivated for more than twenty years and in 2019, the global area under GM crops was 190.4 million hectares, a 112-fold increase since their first commercialization in 1996 [2]. Gains from increased yields and cost savings brought net economic benefits amounting to more than $225 billion and added one hundred million tons to the global crop production without the need for using additional land for cultivation [3]. The development of insect resistant GM crops resulted in reduction of insecticides by 775.4 million kg (8.3%) and decreased the environmental impact of these chemicals by 18.5%. By cutting fuel usage associated with the production of chemical spray runs and tillage, this technology also reduced carbon dioxide emissions equivalent to removing more than 15 million cars from the roads [4]. However, wider adoption of GM crops remains the subject of biosafety concerns due to potential risks such as gene flow, evolution of resistance in insects and weeds, adverse effects on beneficial non-target organisms, or toxicity and allergenicity to humans.
A wide range of pests and pathogens (over 50 insect and about 10 nematode species, 11 viral, 6 bacterial and over 20 fungal pathogens) [5] threaten potato (
Potato is one of the few crops naturally susceptible to infection by agrobacteria, so the first report on the generation of transgenic potato plants using
Isolated in 1901 as the causative agent of silkworm disease,
When insects feed on Bt-plants, ingested Cry protoxin is solubilized and proteolytically activated in the alkaline environment of the insect midgut to the active toxin. The activated toxin goes through complex sequential binding events with an array of receptors on the surface of midgut cells, beginning with binding to cadherin, that facilitates additional protease cleavage and assembly of oligomeric forms of the toxin. The oligomers have increased binding affinity to the secondary receptors, leading to membrane insertion and lytic pore formation [16]. Such midgut tissue disruption halts insect feeding and causes subsequent mortality.
Transgenic potato lines with introduced Cry3A delta-endotoxin from
Expression of several Bt-toxins of Cry1 or Cry9 classes, that target lepidopteran pests, conferred resistance to the potato tuber moth (PTM), a major potato pest in tropical and subtropical regions. Bt-lines with variable level of PTM resistance have been obtained after potato transformation with
Moreover
Insect resistance has become a significant problem after WWII, when intensive agriculture with reliance on chemicals and uniform cultivation practices led to about 17,000 cases of insecticide resistance among 612 insect species by 2020 [9]. Since Bt-crops also provide strong and uniform selection pressure on insect populations it is hard to believe that pest problems can be solved with Bt-approach alone. By 2017, two decades after their commercialization, reduced efficacy of Bt-plants caused by field-evolved resistance has been reported in 16 out of 33 major crop pest populations, compared to only 3 reported in 2005 [31].
In Cry3A-potato, toxin was expressed at a very high level relative to the CPB susceptibility: at least 50 times as necessary to kill first instar, and at least twofold as necessary to stop third and fourth instar development or to arrest adult egg laying [17]. Although effective in short term, this high-dose strategy represents an extremely high selection pressure for developing resistance in the insect populations, and without additional management practices, it has been predicted that CPB can develop resistance to Bt-potato within 6 generations [32]. CPB resistance potential has been demonstrated in the laboratory by repeated Cry3A toxin application, resulting in about 60-fold increase in resistance ratio after 12 generations [33], and about 300-fold increase after 35 generations [34].
Developing Bt-resistance is a complex and diverse process, and populations of the same insect species of different origins may exhibit different mechanisms of resistance to the same Cry toxin [35, 36]. Two major resistance mechanisms are: alteration of midgut proteases involved in processing of Cry proteins in the insect midgut; and modification of binding sites for Bt-toxins. Other resistance mechanisms may include retention of Bt-toxin by the midgut peritrophic membrane, aggregation of toxin proteins by the midgut esterase, elevated melanization activity of the hemolymph and midgut cells, increased rate of repair or replacement of affected epithelial cells, and increased antioxidant activity [37]. Bt-resistant CPB strains exhibit at least two levels of adaptive responses that render immunity to the Cry3A toxin: the first is lower toxin binding to the receptors, probably as a consequence of reduction of binding sites within the receptor or reduction in receptor numbers, while the second one are changes in digestive enzyme profiles and specific increase in aminopeptidase activity [38]. Although this alteration of CPB digestive profile is not connected with toxin processing or its inactivation, it can be involved in modulation and amplification of signals that activate specific innate immune responses such as melanization, coagulation and defense peptide synthesis [39] – mechanisms that have been confirmed in overcoming the exposure to Bt-toxin in other insect species [35, 40].
Moreover, plasticity of its life cycle, large pool of genetic variation in life history traits and capability to effectively cope with naturally occurring host plant toxins or almost every chemical insecticide, leave no doubt that CPB can develop resistance to Bt-potato, given sufficient time. This also brings concerns on whether CPB can be prevented from developing resistance to Bt-potato – since with only a single resistance gene expressed, the high dose/refuge strategy is the only resistance management option available [41]. Although such strategy can hinder accumulation of initially rare homozygous resistance genes in Bt-exposed insect populations by decreasing selection pressure, its effectiveness is questionable in the case of CPB. While the susceptible beetles are “arrested” on Bt-potato, in the resistant strains ingestion of Cry3A toxin significantly increased both CPB larval motility and adult flight activity, whereby more physiologically resistant individuals showed higher behavioral responsiveness. Such behavioral resistance can affect gene flow between susceptible and resistant beetles, increasing distribution of resistant homozygous CPB offsprings within and between Bt-potato fields [33, 42]. In addition, effectiveness of the refuge strategy will be compromised not only when expressed toxin genes do not kill all of the heterozygous progeny, but also if resistance is non-recessive. Evidence of both the laboratory-selected [43] and field-evolved [44] resistance to Cry toxins indicates that some populations of target pests evolve dominant resistance alleles, which can be hardly defeated with the refuge strategy.
When exploring the functions of specific regions of Cry proteins, some of site-directed mutations resulted in increased binding affinity of Cry toxins to insect midgut receptors, conferring additional toxicity. For example, a triple Cry1Ab mutant protein showed up to 36-fold increase in toxicity [45], while multiple Cry3A mutations conferred 2-fold higher toxicity against CPB [46] compared to wild-type Cry toxins. Deletion of small regions of the toxin can result in increased toxicity or in toxins that could counter insect resistance to native Cry toxins. Deletion of 42 residues of the amino-terminal region resulted in an up to 6.6-fold increase in Cry2A toxicity against a lepidopteran pest [47], while Cry1AMod toxins (that due to the lack of α-helix can form oligomers in the absence of cadherin receptor) are effective against Cry1A-resistant target pests with mutations in the cadherin gene [48]. Additionally, added cadherin receptor fragment showed significant synergistic effect with Cry toxins, including 3.7-fold and 6.4-fold enhanced toxicity of Cry3Aa and Cry3Bb, respectively, to CPB [49].
The specificity of Cry proteins allows targeting a single pest or closely related insect species within the same order, but such specificity does not provide a wide range of protection. Improving or broadening the range of protection (as well as minimizing secondary pest infestations upon primary pest control) can be achieved through combining multiple resistance factors – a strategy that at the same time prevents or delays the evolution of insect resistance. The construction of hybrid Cry toxins can confer a wider target spectrum or higher toxicity than each of the parental toxins from which they are derived. Examples include hybrid Cry1Aa/Cry1Ac and Cry1Ab/Cry1C toxins, that exhibited 30- and 10-fold higher toxicity against target pests [50, 51]. Furthermore, a
For instance, concurrent use of one-toxin and pyramided two-toxin crops will enhance resistance to pyramided Bt-plants if the two-gene plants produce a similar toxin as the single-gene plants (for example, this is the case for marketed maize and cotton where the additional Bt-gene was “added” to an already existing Bt-line). Target pests can evolve a single gene resistance that overcomes both Bt genes used in the pyramiding, even if expressed Bt-toxins have different binding sites. A clear example are
In 1999, laboratory studies showed that Bt-maize pollen had deleterious effects on Monarch butterfly larvae [57], raising questions and concerns about Bt-crop impacts on non-target organisms. Additionally, since both target and non-target insect pests ingest toxin when feeding on Bt-plants, Bt-toxin may also affect beneficial predatory arthropods through consumption of target pests or by facultative feeding on transformed plants.
Riddick and Barbosa [58] showed no adverse effect on survival, fitness or predation potential of
Furthermore, studies on commercialized SmartStax maize with six Bt-genes (
Although free Bt-toxin released in root exudates and from decaying plant residues is rapidly degraded by soil microbes, it can be stabilized by binding on clays or humic substances and stay unchanged for two weeks to 6 months [67], depending on soil composition and pH, or crop species [68]. However, studies on Bt-crops have generally revealed no or minor transient effect on earthworms, nematodes, protozoans, bacteria, and fungi in soil [68].
Due to the acidic environment of the mammalian digestive tract and the absence of specific receptors, it is generally accepted that Bt-toxins do not bear substantial risk for human health. Additionally, about 60 years of history of using Bt-products as biopesticides showed that risks of toxicity or allergenic reactions to the Cry proteins are minimal. Cry3A toxin does not exhibit acute oral toxicity to mammalians in doses 10,000 times higher than its amount in potato tubers, and is rapidly digested
As a reflection of more than one hundred million years of coevolutionary “arms race”, plants developed numerous mechanisms to resist the attacks of pathogens and herbivores. Here, being part of the plant “chemical warfare” arsenal, secondary metabolites take an important place, with more than 200,000 known compounds with defensive activity. Among that broad repertoire, protein antimetabolites such as lectins, α-amylase inhibitors and especially plant protease inhibitors (PIs) are the most used for engineering crop resistance against various pests.
The most important role of PIs in plants is protection from both biotic and abiotic stresses. They may also have other functions: from tissue-specific regulation of endogenous proteases – especially in storage organs such as seeds and tubers [75], to the regulation of programmed cell death [76]. About 500 plant PIs were described, and according to the protease type they inhibit, PIs are classified as cysteine, serine, aspartyl and metallo protease inhibitors [77]. Generally, the inhibition is based on PIs binding to or near the enzyme active site, forming a stable complex with a low dissociation constant. This complex is often additionally “locked” by disulphide bonds, so that upon eventual hydrolysis the inhibitor remains associated to the enzyme, effectively blocking access of the substrate [78]. The mechanism of PIs antimetabolic effect on insects has not been fully elucidated and, due to its high specificity, it is assumed that different types of PIs also have different modes of action. The simplest model implies a direct antidigestive effect due to inhibition of proteolysis [79]. The second, more accepted model, is based on compensation for the loss of proteolytic activity – proteinase hyperproduction – which by redirecting amino acid utilization reduces their availability for insect growth and development [80] which, in addition to reduced performance, often increases insect mortality. PIs can also disrupt processes such as molting, neuropeptide synthesis, water balance, and enzyme regulation [81, 82, 83] or directly interfere with insect reproductive processes [84].
The early evidence on the protective role of PIs came in mid-20th century, when it was observed that soybean products negatively affect development of red flour beetle larvae [85]. In a pioneering research, Green and Ryan [86] reported on a rapid, both local and systemic, accumulation of PIs in potato and tomato leaves upon CPB attack, demonstrating the importance of PIs in plant defense against insects. Not long after, the first PI-transformed plant, tobacco expressing cowpea trypsin inhibitor, CpTI, conferred increased resistance to several lepidopteran, coleopteran and orthopteran insect pests [87]. This initial success triggered a generation of numerous transgenic plants expressing different PIs, more or less efficient in control of target pests. However, despite this promising development, none of PI-transgenic plants have been commercialized to date. One of the reasons is the conclusive “acute mortality” efficacy of Bt-plants, similar to the chemical insecticides. By contrast, PIs often cause decrease in insect fitness on a relative level, such as a reduction in growth and reproduction or extended development, that in a time scale can significantly reduce the size of pest population (for example, prolonged larval development brings longer exposition to predators, while the reduction in body mass decreases investment in reproduction). Secondly, a more important reason are adaptive capacities of insects that can compromise this approach, clearly demonstrated in some cases. These evolutionary, diet-induced strategies include overproduction of sensitive digestive enzymes that outnumber inhibitors, switching to digestive protease complements insensitive to PI or PI degradation with non-target proteases [88].
After evidence of deleterious effects of E-64, a broad spectrum thiole cysteine PI isolated from
These results clearly demonstrate that, due to its exceptional adaptability to the different host plant protective compounds [88], CPB can hardly be controlled by a single, narrow spectrum PI. Thus, to achieve more efficient control and prevent compensatory insect responses, broadening the spectrum of inhibition by protein fusion, transgene stacking or using multidomain PIs appeared as a possible solution. However, only a slight reduction in CPB larvae performance was achieved in potato expressing stacked rice cystatins, OCI and OCII [94, 95] or with multidomain serine PI from locust (LIP), active against both trypsin and chymotrypsin [96].
In contrast to this, equistatin, a PI from the sea anemone, with one domain that inhibits cysteine and a second domain active against aspartic proteases, had detrimental effect on CPB larvae growth and significantly increased their mortality after ingestion of equistatin-coated potato leaves [97]. Unfortunately, with expression of this potent PI in potato very low resistance level against CPB was achieved: the amount of active inhibitor in leaves was considerably reduced due to its degradation by native potato proteinases [98]. The promising results came with a hybrid CDI-CCII inhibitor (fusion of CDI with maize cystatin II), also active against both aspartate and cysteine proteinases. When painted on potato leaves, CDI-CCII initially reduced CPB larvae growth and food consumption by about 50% [99], but its real effects still remain to be proved in long-term feeding assays. Finally, fungal cysteine PIs, macrocypin and clitocypin, emerged as more favorable. Exhibiting strong inhibition of CPB cysteine proteinases, these PIs, introduced in potato, reduced growth and increased development time of CPB larvae [100, 101]. Moreover, the most promising trait of macrocypin and clitocypin is the absence of CPB digestive compensatory responses [100, 101] observed for PIs derived from other sources. However, relatively low expression was achieved in transgenic potato and, since they act in dose dependent manner, it is necessary to improve macrocypin and clitocypin expression levels for more pronounced negative effects on CPB larvae.
Additionally, potato expressing serine PI (CpTI or Soybean Kunitz, C-II and PI-IV) exhibited enhanced resistance to the lepidopteran larvae with about 50% reduction in total insect biomass [82, 102].
Several approaches based on structure–function models have been used to improve the inhibitory potency of protease inhibitors against specific proteases, including site directed mutagenesis of specific amino acids, molecular phage display procedures involving random mutagenesis in specific regions of the inhibitor sequence, or activity-based functional proteomics approach. By single mutations at the positively selected amino acid sites of the tomato multicystatin SlCYS8, variants with improved inhibitory potency toward the CPB digestive proteases were generated [103], and functional proteomics approach was used for identifying variants that efficiently capture CPB digestive protease targets [104]. P2V10, the most potent variant of SlCYS8 PI, expressed in potato, significantly reduced growth of CPB larvae in a 72 h feeding assay [104]. Similarly, after 4 days of feeding on potato expressing a modified variant of cystatin from barley (HvCPI-1 C68 fi G), that targets the cathepsin B-like fraction of cysteine digestive proteolysis, CPB larvae had about 23% lower weight, probably due the metabolic cost associated with the hyperproduction of inhibited digestive proteases [105]. However, knowing the remarkable CPB larvae adaptability to adjusting their digestive profile to functionally distinct plant PIs, studies assessing the long-term detrimental effects of these engineered cystatins are needed.
Although the usefulness of recombinant PIs expressed alone still remains to be proved or improved, they can enhance Cry toxicity. Several serine protease inhibitors can increase the insecticidal activity of Cry toxins 2–20 fold [106] and delay the resistance evolution of the targeted pest [107]. Although it is not known how PIs enhance Bt-toxin activity, it is supposed that they may inhibit the inactivation of Bt-toxins by specific gut proteases, or prevent the degradation of membrane receptors, increasing binding ability of Cry toxins [108]. In such way, hybrid SN19 (
Due to the existence of targets in most organisms in nature, beside the toxic effect on the pest, recombinant PIs can directly affect the digestive proteolysis in pollinators, symbionts and/or indirectly, through prey feeding on transgenic plants, they can endanger the ecological function of predators. However, although artificial diet studies indicate that predatory insects may be susceptible to the PI, prey-mediated effects are usually not observed when cystatins or CpTI are expressed in transgenic potato. When
On the other hand, although the effects of native plant PIs, such as CpTI or OCI, on non-target organisms have been well documented, there is little evidence of effects of new-generation inhibitors with stronger effects on pest proteinases, hybrid inhibitors or combined effects of several different insecticidal proteins. The challenge, of course, is to find or devise those variants of PIs that show increased activity against the target pest proteinases and decreased activity against proteinases of the host plant or of beneficial insects. Also, cystatins that occur naturally in seeds of rice and maize, present in potato tubers or in egg-white, are not novel in the human diet, and expressed in transgenic plants should not cause public concerns [114] – but the expression of strong broad-spectrum aspartate and serine PIs may raise many questions in the future.
Widely distributed in nature, lectins are a heterogeneous group of sugar-binding proteins with numerous biological functions. In plants they are involved in the transport and utilization of carbohydrates, cell organization, division and signaling, embryomorphogenesis, phagocytosis or as mediators of plant-microorganism symbiosis [115]. However, their most distinctive role is in plant defense mechanisms against pathogens and pests. Binding to a variety of glycoproteins, plant lectins can inhibit absorption of nutrients by disruption of insect gut epithelium structure or, by interacting with targets in insect hemolymph, fat tissue and ovaries, interfere with a number of physiological processes, such as growth, development and detoxification [116]. Although they can exhibit protective roles against insect pests from different orders, lectins are particularly useful for controlling Hemiptera, that are generally less sensitive to Bt or PIs.
Snowdrop mannose-binding lectin (
However, lectins can negatively impact beneficial non-target organisms, and for instance, preys that were fed on GNA potato were less favored or resulted in smaller, shorter-lived predators or parasitoids [123, 124]. Although they are present in most plants – especially abundant in cereal and legume seeds or potato tubers – lectins are generally considered toxic to animals and humans. So even though GNA did not show considerable toxicity in rat feeding studies [125], there is no doubt that food expressing such proteins requires long-term studies to evaluate its potentially harmful effects.
After the Nobel prized discovery of RNA interference (RNAi) as a basic mechanism of post-transcriptional gene silencing by double-stranded RNA (dsRNA) [126] RNAi has become a powerful experimental tool for determining gene functions, had an immense impact on biomedical research and found its application in the management of insect pests. Evolutionarily conserved in all eukaryotes, the mechanism of RNAi is involved in different processes including internal gene regulation (micro RNA or miRNA pathway), genome protection against transposons (piwi-interacting RNA or piRNA pathway) and defence against viral infections (small interfering RNA or siRNA pathway) [127]. Althought the siRNA pathway in insects mostly represents the first line of defense against viral RNA, it can be exploited for introduction of specific dsRNA that, through mechanism of RNAi, can initiate degradation of complementary endogenous insect mRNA. Thus, selection of any target gene and delivery of its sequence-specific dsRNA to cells can lead to functional knockout of that gene – affecting insect growth and development or increasing their mortality. A first proof-of-concept came in 2007, when transgenic maize expressing V-ATPase-specific dsRNA showed significant reduction in feeding damage caused by western corn rootworm (WCR) [128]. Maize with dsRNA transcript containing a 240 bp fragment of the WCR
However, various studies showed that different insect orders differently respond to orally delivered dsRNA – coleopterans are mostly sensitive, while RNAi efficiency is low for most lepidopterans. Multiple mechanisms contribute to this variability, including instability of dsRNA upon ingestion, insufficient dsRNA internalization, endosomal entrapment, deficient function of the RNAi machinery and reduced systemic spreading. Once consumed, the dsRNA first has to avoid degradation by dsRNases (dsRNA-specific ribonucleases) on their way through insect digestive tract. Level of dsRNA degradation by saliva or midgut nucleases varies among different insect orders and, for instance, midgut stability of dsRNA is greater in the CPB (Coleoptera) than in
Once taken up in the cytoplasm, dsRNA is recognized by the core RNAi machinery and processed into 21–23 bp siRNA by the enzyme Dicer 2 (DCR-2). The siRNA are loaded onto Argonaute 2 (Ago-2) protein and incorporated into the RNA-induced Silencing Complex (RISC). Upon degradation of the passenger strand of siRNA, RNase active domain of Ago-2 cleaves the mRNA recognized by the siRNA guide strand, inducing gene silencing. One of the reasons for efficient RNAi in coleopterans is the duplication of core RNAi pathway genes, including DCR-2 and Ago-2 [133]. Additionally, in CPB, components of miRNA and piRNA pathways are also critical for effectiveness of gene silencing by the siRNA pathway, but their involvement in dsRNA-mediated RNAi needs to be further investigated in Coleoptera and other insects [134]. A particularly interesting aspect of the RNAi response in insects is its potential systemic character, whereby the silencing signal can spread from the midgut to other tissues, causing systemic RNAi. The exact nature of this signaling pathway still remains elusive, and efficient silencing of genes in midgut tissue was predominant, especially in more derived dipteran and lepidopteran species that appear to be more refractory to systemic RNAi [135].
Although there is a vast number of essential genes in insect genomes, the choice of the target gene can significantly affect the efficiency of RNAi – but the factors making one essential gene a better target than another one are not currently understood. Variation in transcriptional activity, mechanisms of expression regulation, mRNA stability and its accumulation level may play an important role in defining a particular gene susceptibility to dsRNA, and screening of a larger number of potential target genes for RNAi efficiency remains the only reliable method of choice.
The availability of the CPB transcriptome [136] allows specific targeting of CPB genes critical for normal physiological processes and numerous studies demonstrated successful knockdown of target genes in dsRNA-fed CPB. Silencing the expression of genes that are crucial for maintaining physiological functions, such as actin and V-ATPase genes, or genes coding components involved in protein transportation (
Reduction in CPB juvenile hormone (JH) titer, that regulates metamorphosis and reproduction in insects, was achieved by knockdown of JHAMT (JH acid methyltransferase), the last rate-limiting enzyme in JH biosynthesis. Feeding on transgenic potato plants expressing ds
On the other hand, transformation of chloroplast DNA has potential for overcoming the constraints of nuclear transformation in dsRNA-mediated pest control. First advantage of transplastomic plants are markedly high gene expression levels, that due to tissue specificity, occur predominantly where functional plastids are present. An example is expression of Cry2Aa2 protoxin in tobacco chloroplasts in 20- to 30-fold higher levels than current commercial nuclear transgenic plants, which is lethal for both susceptible and Bt-resistant target insects [149]. Secondly, a great advantage of plastid transformation is the stability of dsRNA in plastids, as chloroplasts do not have the RNAi machinery. Among about 130 genes encoded by the chloroplast genome, none is Dicer-like or Argonaute-like, and there is no evidence of import of these nuclear-encoded proteins in chloroplasts [150]. Three recent studies demonstrated that when expressed from chloroplast genome, hp/dsRNA can confer a high level of protection against either lepidopteran (
Furthermore, chloroplast genome transformation also offers other advantages over nuclear transformation, including introduction of multiple genes in a single transformation event and lack of gene silencing, position or pleiotropic effects. Additionally, maternal inheritance excludes plastid genes and therefore reduces dispersion of the transgene by pollen transmission, increasing the biosafety of transgenic plants. However, plastid transformation is still much more challenging than nuclear transformation and limited by the methods of DNA delivery, homologous recombination efficiency and the methods for efficient selection and regeneration of transformants [154].
Numerous studies have shown that under long-term pressure of control strategies such as chemical insecticides or Bt-toxin, insects can rapidly evolve resistance, and there is no reason to believe that it would be differently with RNAi. Theoretically, there are three possible sources of resistance: mutations in the sequence of the target gene, mutations inactivating the RNAi machinery and mutations that affect the stability and/or uptake of ingested dsRNAs in the insect digestive tract. First two mechanisms are unlikely to become source of resistance. For instance, in CPB the mismatch rate of
With perfect sequence homology between dsRNA and mRNA only target gene suppression is expected, but it appears that siRNAs operate within cells with a certain level of “freedom” among targets. Mutation analyses showed that RNAi can be efficiently triggered with >80% sequence identity between siRNA and mRNA [158] but this mismatching tolerance can vary with insect species, target gene and dsRNA concentration [159, 160]. Moreover, dsRNA can provoke responses independently of its sequence, affecting insect antiviral immunity, gene expression and performance [158, 160]. Although not fully understood, these effects are particularly pronounced for dsRNA administered at high concentrations, supposing that high levels of siRNA may saturate the core RNAi machinery [161]. Given the small sizes of siRNAs, off-target effects that can appear in RNAi are probably quite common [162] and not considered as a concern in target organisms, but off-target binding in non-target organisms can represent a hazard if they are sufficiently exposed to the RNAi. To date, question how dsRNAs affect target and off-target genes in non-target organisms has received little attention, and existing studies indicate that the insecticidal effects of
dsRNAs exhibit low persistence in environment and microbial degradation of nucleic acids has been shown to be a key driver for such lack of stability. Biological activity of
Vertebrates are exposed to dietary intake of a number of various dsRNAs from animal, plant or microbial origin. Some are completely complementary to human or animal genes [168] and capable of initiating the RNAi pathway if they reach a target cell. However, there are numerous biological barriers, including nucleases along the digestive tract, and in bloodstream, series of cellular membrane barriers and endosomes significantly reduce dsRNAs to the levels insufficient for mediating RNAi. In 28-day repeat oral toxicity study in mice with
Gene flow is the transfer of genetic material from one organism to another, including inheritance (vertical gene transfer) or transfer between unrelated species (horizontal gene transfer). Althought horizontal gene transfer can contribute in “shaping” genomes of both prokaryotes and eukaryotes, there are almost no evolutionary examples of gene transfer from eukaryotes to bacteria [171]. Transfer of plant DNA to bacteria has been demonstrated at a very low frequency under artificial conditions, and the only genes from GM plants that are likely to be successfully transferred are other bacterial genes, commonly used for selection in transformation [172]. More than 90% of transgenic plants that have been generated in different laboratories carry one of the three genes used for selection (resistance to antibiotics kanamycin or hygromycin, or herbicide phosphinothricin) [173], all of bacterial origin. Antibiotics are the most effective selection system for potato transformation, increasing its efficiency from 0.2%–4.5% under non-selective conditions to over 80% [174, 175]. However, they generally have no use after the selection phase of transformation, and can be completely removed or excised by different approaches, including segregation from the gene of interest after co-transformation, and different site-specific or homologous recombination systems [176]. In this way, using self-crossing segregation or inducible self-excision by the Cre-loxP system, selectable marker-free transgenic potato lines with increased resistance to pest or pathogens were created [177, 178], alleviating possibility of horizontal gene transfer.
On the other hand, vertical gene transfer, especially mediated by pollen, raises more concern. Transgene escapes have been documented for cotton, maize, soybean, oilseed rape, rice and wheat, indicating global dimensions of this problem [179]. In the case of Bt-plants, crop-to-crop gene flow can cause seed contamination, decrease efficiency of refuge strategies, or interfere with conventional or organic crop production. For instance, in Mexico where GM maize was not allowed for commercial cultivation, transgene escapes (
Cross-pollination between GM and non-GM potato should be less worrying, since vegetative propagation by tubers (rather than true seeds) is the dominant reproduction strategy of potato, and tubers are not affected by the plant fertilization with “foreign” pollen. Outcrossing has been observed to occur only between adjacent potato fields, with rapid decreasing rate with distance, and no cross-pollination detection when the pollen-receiving plants were separated by more than 20 meters from the GM plants [182]. Additionally, majority of modern cultivars that evolved from complex hybridizations among several diploid and polyploid potato species, suffer from different types of male sterility and produce little or no viable pollen. Also,
Crop improvement by genetic engineering requires obtaining transgenic lines with adequate expression of the heterologous gene and simultaneous preservation of all elite parental genetic attributes. One of the main limitations in achieving these requirements is the emergence of atypical plants – most often as a result of insertional mutagenesis or somaclonal variations that may occur in the tissue culture itself and/or during transformation.
In many plant species, including potato [187], the frequency of heterologous DNA insertions within coding or regulatory gene sequences exceeds 50% upon genetic transformation. Additionally, insertion-site mutations can alter the expression patterns of neighboring genes, especially if the heterologous gene is under the control of a strong promoter [188]. Another type of mutation, related to the transformation process itself, can occur in any part of the plant genome (genome-wide mutations) and is reflected in DNA polymorphism between transgenic and non-transgenic plants [189]. These latter changes are of epigenetic nature: the transformation process can activate transposon elements (TEs – whose activity is normally prevented by DNA hypermethylation), which then increase mutation rates and genomic rearrangements [190]. It is assumed that the same mechanism – activation of TEs – underlies somaclonal variations, a phenomenon associated with
Insertional mutagenesis is not expected to be manifested in potato, being autotetraploid and possessing three other alleles that can potentially compensate for the insertional effect of a gene functional deletion. Even when insertional mutagenesis produces visible phenotypic changes due to the high heterozygosity of commercial potato cultivars, such phenomena are considered an extremely rare event [192]. On the other hand potato is quite susceptible to somaclonal variations in tissue culture even in the absence of transformation [193]. The incidence of atypical plants attributed to somaclonal variations, ranges between 15% and 80% in the population of transgenic potato lines, depending on cultivar [192, 194]. These are often manifested as reduced growth, deformed leaf shape, lower yield and other changes in development, clearly visible in changing field conditions, rather than in uniform ones such as greenhouses or
Beside insertional mutagenesis or somaclonal variations, the unexpected changes in transgenic lines may be a consequence of the transgene expression itself. It is especially expected with PIs, that may interact with plant endogenous protease targets structurally and functionally related to insect digestive proteases, bringing both positive and negative pleiotropic effects
Unintended traits have been identified in commercial GM crops, including insect or herbicide resistant maize, cotton, soybean and oilseed rape – that can exhibit different agronomic and compositional changes relative to their non-GM parental lines [201]. For example, Mon810 maize, carrying
Owing to public concern and reserved acceptance of transgenic crops in many parts of the world, two approaches, cisgenesis and intragenesis, are designed as an alternative to “old” transgene technology. Both concepts include introduction of genetic material derived from the species itself (intragenesis) or closely related, cross compatible species (cisgenesis). Although they use a genetic transformation step, the modified crop genome is designed to not contain any foreign gene, including selectable markers. Therefore, crops developed using these techniques correspond to plants generated through conventional breeding, but without unintentional introduction of undesired genetic elements. Intragenesis has been successfully used for developing potato with high amylopectin content by silencing of the granule-bound starch synthase gene,
Genome editing is the latest and most potent molecular technology. Using programmable endonucleases (Zincfinger, TALENs or CRISPR-Cas), alterations can be made at precise locations in the genome, including targeted insertion, replacement or disruption of genes in plants. Because of their precision, these techniques can produce fewer unintended effects, and therefore “edited” crops are considered potentially safer than those generated by random mutagenesis or insertion. In case of potato, both TALENs and CRISPR/Cas9 technologies have been mainly used to improve tuber quality (glycoalkaloids reduction, low acrylamide content and altered starch metabolism) or for herbicide resistance [211], but despite unlimited potential in genetic engineering, no pest-resistance gene incorporation has been reported yet. Importantly, CRISPR-based gene drives could be implemented to spread desirable genetic elements through pest populations themselves. For CPB, there is only one such report to date, where CRISPR/Cas9 was used for
For the growing world population that is expected to reach 10 billion by 2050, food production should be increased by 25–70% and, at the same time, it is necessary to reduce nutritional losses, greenhouse gas emissions from agriculture, pesticide overuse and address other environmental concerns [213].
Potato is now the world’s third most important crop for human food consumption, after wheat and rice, but its production in the last 10 years stopped between 360 and 370 million tons annually [214]. Additionally, yield potential of potato has remained relatively unchanged, despite intensive breeding efforts [215], and century-old varieties (i.e., Russet Burbank and Bintje) are still cultivated due to lack of significant genetic improvements in potato. Narrow genetic base as a result of clonal propagation, multiple constraints such as inbreeding depression, self-incompatibility and incorporation of undesirable traits, limit the progress in conventional development of inbred potato lines [216]. On the other hand, genetic engineering has shown potential for fast, feasible, economic and environment-friendly introduction of resistance (and other beneficial) traits in commercially grown crops. However, to make full use of that potential it is necessary to improve existing and bring about new, more sustainable and cleaner gene manipulation technologies. By optimization of transgene expression level, its temporal or spatial programming (i.e., by use of wound-inducible or tissue-specific promoters), generating marker-free modified plants and exploiting new approaches such as cisgenesis/intragenesis or genome editing – it is possible to both decrease unintended effects and increase efficiency and public acceptability of transgenic crops.
For potato, there are no GM varieties with insect resistance traits in the markets, and strategies that rely on insecticides cannot be avoided – as well as their failure in pest control. For instance, imidacloprid, a neonicotinoid successfully used for almost 10 years in CPB control, started being ineffective at the beginning of this century [217]. On the other hand, as global population continues to expand, food production, including potato, has to increase by many folds and with wild potato varieties as only source of resistance traits and their introduction by breeding, that seems unattainable. Also it is questionable whether all potato pests, CPB especially, could be stopped by resistance factors existing in
As a crucial concept of insect resistance management, refuges are essential for durability of both stacked and single-toxin crops, and where resistance is rare, 20% (or at least 10% for stacked traits) of a pest host plant refuge may be sufficient to delay resistance by a decade or more [31]. Smaller refuges are insecure even under highly effective toxins (or other traits) and all cases of field-evolved resistance are associated with low refuge presence, as one of the main factors [221]. Additionally, within IPM context, refuges also provide better support to populations of natural enemies, that are not only important in target pest control, but to prevent non-target secondary pest outbreaks that can seriously reduce benefits from introduced traits and bring production back to running on the insecticide treadmill [222]. Adding pheromone disruption, mass trapping or intercropping arrangements – integrated into scientifically supported management and adapted to the pest biology – can efficiently reduce pest population size, keeping damage below the economical threshold. Experiences with combination of the simplest practices in potato fields in some parts of the USA, such as rational use of chemical insecticides, trap rows and crop rotation [223] proved a potential of well-structured IPM approach to balance one technology with other complementary strategies. Such avoidance of relying on only one means of control would require complex pest adaptations that are less likely to happen compared to the occurrence and fixation of random single gene mutations that can render resistance to insecticides, Bt-toxin, PIs, RNAi or any other measure that may be implemented in the future.
The benefits of pest-resistant GM crops, incorporated in well-balanced IPM strategies, are clear – but it is also necessary to define and understand their limitations and risks. Heavy dependence and overuse of insecticides undoubtedly had many consequences: food poisoning, reduction in biodiversity, negative effects on non-target species and other formidable impacts on environment – and genetic engineering provides a chance to not repeat all those mistakes. However, we cannot expect that Bt or other pest-resistant modified crops will not have long-term ecological or evolutionary consequences, as well as that small or substantial compositional changes, as intended (or unintended) quantitative or qualitative alterations of metabolites, nutrients or toxins, cannot impact ecological interactions and/or food or feed safety. Such risks are present and inevitable, can vary depending on traits introduced and strategy used for its introduction – and with a precautionary approach, at least some of them can be avoided or mitigated. Additionally, every generated crop line is created in a unique event and should be evaluated for risks, benefits and sustainability only on a case-by-case basis.
So, taking all together, is genetic modification of plants a thrift or a threat? It only depends on how carefully and advisedly we use that tool in our hands.
This work was supported by the Ministry of Education, Science and Technological Development of the Republic of Serbia, contract number 451-03-9/2021-14/200007.
The authors declare that they have no conflict of interest.
Uropathogenic
Adhesins are adhesive organelles, notably fimbriae, that promote bacterial colonization. Some adhesins also promote bacterial invasion of the host cell. Adhesins are thought to be the most important virulence-associated molecules which function in UPEC pathogenicity. The adhesins can also directly trigger host and bacterial cell signaling pathways. They can also facilitate the delivery of other bacterial products to the host tissues [17]. Prominent bacterial cell surface virulence factors, which play significant roles in UPEC pathogenicity include type 1 fimbriae [11]; Class I, Class II, and Class III of P-fimbriae [18, 19, 20]; Dr. family of adhesins for binding to the decay-accelerating factor (DAF) [21]; Curli fimbriae which functions as binding factor and biofilm producer [22]; and S-fimbriae [14, 23, 24]. Type 1 fimbriae have the most significant effects in UTIs as they enhance bacterial survival and growth, enhance inflammatory reaction at the mucosa, bacterial invasion, and control biofilm production [7]. P-fimbriae have the second most prominent role in UPEC-associated pathogenesis of human ascending UTIs and pyelonephritis. They promote UPEC adherence to the matrix of the mucosa and tissues and trigger cytokine production [25, 26, 27, 28, 29, 30].
UPEC secrete several virulence toxins which are responsible for the damage of the host cells and host inflammatory response. α-hemolysin (HlyA) is the most virulent toxin produced by UPEC. The effects of HlyA in UTIs are dependent on its dosage produced by UPEC. At high concentration, HlyA destroys the erythrocytes and allow UPEC to break through the mucosal barriers, damage immune system, and depletes iron stores of the host [31, 32, 33, 34]. At low concentration, HlyA induces cell death in the bladder using proinflammatorycaspase-1/caspase-4. This causes kidney damage and scarring; oscillations of Ca2+; ascension and colonization of ureters and kidney parenchyma in the renal tubule epithelia resulting in the disruption of normal flow of urine [35, 36, 37, 38]. The stimulation of
Urinary tract has limited iron. However, UPEC are able to produce small iron chelator molecules, known as siderophores, to scavenge ferric iron (Fe3+) in the host. The most prominent ones are yersiniabactin, salmochelin, and aerobactin [48, 49]. The yersiniabactin and its receptor, FyuA, are encoded in a PAI [50, 51]. It has also been reported that for efficient biofilm formation by UPEC, FyuA is required [52]. UPEC also secretes another important hydroxamate siderophore called aerobactin. This is produced from the condensation of two lysine and a citrate molecules. During UPEC invasion, the bacterium secretes salmochelin. Its outer membrane siderophore receptor (IroN) transports different catechol siderophores, including N-(2,3-dihydroxybenzoy)-L-serine and enterochelin also called enterobactin [53]. Enterobactin has less solubility and stability than aerobactin [54, 55, 56] but has higher iron affinity than aerobactin in aqueous [55, 57]. UPEC also uses enterobactin for Fe3+ scavenging in the urinary tract [9]. However, enterobactin can be inactivated by the host proteins such as serum albumin and siderocalin thereby preventing its uptake [58]. UPEC overcomes this instability by modifying the enterobactin to salmochelin by glucosylation through the enzymatic action of glucosyltransferase and prevents it from being recognized by the host proteins [9]. Also, UPEC has another iron acquisition system called haemin uptake system consisting of Ton-B dependent receptor (ChuA) and heavy metal associated (
UPEC-associated fitness and virulence. Adapted from the work by Servin [
Lipopolysaccharide (LPS) is a major part of the cell wall which has highly conserved lipid A-core and repeating O-antigen subunits which vary in different strains of
Capsule is made up of polysaccharides and it covers and protects UPEC from various harsh environmental conditions [66]. The capsule helps UPEC to resist phagocytosis and bactericidal effects of complements in the host. It also confers antimicrobial resistance and antiserum activity to UPEC [54, 61]. Capsules like K1 and K5 interfere with the proper response of the humoral immunity of the infected host [66]. The K1 polysaccharide plays a significant role in intracellular bacterial community (IBC) development and the pathogenesis of several UTI stages [54, 67].
Toll receptor (TIR)/interleukin1 (IL-1) receptor domain-containing protein (TcpC) is a novel class of virulence factors that destabilize TIR signaling for UPEC to survive during UTIs [68]. Interaction of TcpC with myeloid differentiation primary response 88 (MyD88) found in the host ends the downstream signaling pathways mediated by TLRs [69].
UPEC produces outer membrane protease T (OmpT) that catalyzes plasminogen activation to plasmin [70]. OmpT helps UPEC to persist in the urinary tract when protamine and other cation peptides cleave with antibiotic activity [71, 72]. UPEC also decreases cytokines production by blocking nuclear factor kappa-light-chain-enhancer of activated B cells (NF-ĸB) [68]. In Table 1, prominent UPEC virulence factors, their role and genetic markers are presented.
Virulence factor | Role | Genetic markers/gene name | References |
---|---|---|---|
Afimbrial adhesions | Binding factor | [23, 24, 54] | |
Cytotoxic necrotizing factor 1 | Toxin | [38, 39] | |
Curlifimbriae | Binding factor | [22] | |
Dr family of adhesions | Binding factor | [21] | |
Haemin | Iron uptake and biofilm formation | [59, 60, 61] | |
Type 1 fimbriae | Binding factor | [8] | |
Ferric yersiniabactin uptake receptor | Iron uptake and biofilm formation | [62] | |
α-hemolysin | Lyses red blood cells | [33] | |
Salmochelin | Siderophore receptor | [51] | |
Aerobactin | Iron chelation and uptake | [50] | |
Outer membrane protease T | Outer membrane protease production to degrade protamine peptides | [73, 74] | |
Uropathogen specific protein | Movement of UPEC from the urinary tract to the bloodstream | [42] | |
Class I, Class II, and Class III P-fimbriae | For binding to the uroepithelial cells | [18, 20, 21] | |
Serine-protease autotransporter toxin | Vacuolation and tissue damage | [73, 74] | |
S-fimbrial family | Binding factor | [8, 23, 24] |
Virulence factors of uropathogenic
Apart from possessing virulence factors, for the medical importance of
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Radiotherapy and Nuclear Medicine Technology has always been my aspiration and my life. As years passed I accumulated a tremendous amount of skills and knowledge in Radiotherapy and Nuclear Medicine, Conventional Radiology, Radiation Protection, Bioinformatics Technology, PACS, Image processing, clinically and lecturing that will enable me to provide a valuable service to the community as a Researcher and Consultant in this field. My method of translating this into day to day in clinical practice is non-exhaustible and my habit of exchanging knowledge and expertise with others in those fields is the code and secret of success.",institutionString:null,institution:{name:"Majmaah University",country:{name:"Saudi Arabia"}}},{id:"313277",title:"Dr.",name:"Bartłomiej",middleName:null,surname:"Płaczek",slug:"bartlomiej-placzek",fullName:"Bartłomiej Płaczek",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/313277/images/system/313277.jpg",biography:"Bartłomiej Płaczek, MSc (2002), Ph.D. (2005), Habilitation (2016), is a professor at the University of Silesia, Institute of Computer Science, Poland, and an expert from the National Centre for Research and Development. His research interests include sensor networks, smart sensors, intelligent systems, and image processing with applications in healthcare and medicine. He is the author or co-author of more than seventy papers in peer-reviewed journals and conferences as well as the co-author of several books. He serves as a reviewer for many scientific journals, international conferences, and research foundations. Since 2010, Dr. Placzek has been a reviewer of grants and projects (including EU projects) in the field of information technologies.",institutionString:"University of Silesia",institution:{name:"University of Silesia",country:{name:"Poland"}}},{id:"35000",title:"Prof.",name:"Ulrich H.P",middleName:"H.P.",surname:"Fischer",slug:"ulrich-h.p-fischer",fullName:"Ulrich H.P Fischer",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/35000/images/3052_n.jpg",biography:"Academic and Professional Background\nUlrich H. P. has Diploma and PhD degrees in Physics from the Free University Berlin, Germany. He has been working on research positions in the Heinrich-Hertz-Institute in Germany. Several international research projects has been performed with European partners from France, Netherlands, Norway and the UK. He is currently Professor of Communications Systems at the Harz University of Applied Sciences, Germany.\n\nPublications and Publishing\nHe has edited one book, a special interest book about ‘Optoelectronic Packaging’ (VDE, Berlin, Germany), and has published over 100 papers and is owner of several international patents for WDM over POF key elements.\n\nKey Research and Consulting Interests\nUlrich’s research activity has always been related to Spectroscopy and Optical Communications Technology. Specific current interests include the validation of complex instruments, and the application of VR technology to the development and testing of measurement systems. He has been reviewer for several publications of the Optical Society of America\\'s including Photonics Technology Letters and Applied Optics.\n\nPersonal Interests\nThese include motor cycling in a very relaxed manner and performing martial arts.",institutionString:null,institution:{name:"Charité",country:{name:"Germany"}}},{id:"341622",title:"Ph.D.",name:"Eduardo",middleName:null,surname:"Rojas Alvarez",slug:"eduardo-rojas-alvarez",fullName:"Eduardo Rojas Alvarez",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/341622/images/15892_n.jpg",biography:null,institutionString:null,institution:{name:"University of Cuenca",country:{name:"Ecuador"}}},{id:"215610",title:"Prof.",name:"Muhammad",middleName:null,surname:"Sarfraz",slug:"muhammad-sarfraz",fullName:"Muhammad Sarfraz",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/215610/images/system/215610.jpeg",biography:"Muhammad Sarfraz is a professor in the Department of Information Science, Kuwait University. His research interests include computer graphics, computer vision, image processing, machine learning, pattern recognition, soft computing, data science, intelligent systems, information technology, and information systems. Prof. Sarfraz has been a keynote/invited speaker on various platforms around the globe. He has advised various students for their MSc and Ph.D. theses. He has published more than 400 publications as books, journal articles, and conference papers. He is a member of various professional societies and a chair and member of the International Advisory Committees and Organizing Committees of various international conferences. Prof. Sarfraz is also an editor-in-chief and editor of various international journals.",institutionString:"Kuwait University",institution:{name:"Kuwait University",country:{name:"Kuwait"}}},{id:"32650",title:"Prof.",name:"Lukas",middleName:"Willem",surname:"Snyman",slug:"lukas-snyman",fullName:"Lukas Snyman",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/32650/images/4136_n.jpg",biography:"Lukas Willem Snyman received his basic education at primary and high schools in South Africa, Eastern Cape. He enrolled at today's Nelson Metropolitan University and graduated from this university with a BSc in Physics and Mathematics, B.Sc Honors in Physics, MSc in Semiconductor Physics, and a Ph.D. in Semiconductor Physics in 1987. After his studies, he chose an academic career and devoted his energy to the teaching of physics to first, second, and third-year students. After positions as a lecturer at the University of Port Elizabeth, he accepted a position as Associate Professor at the University of Pretoria, South Africa.\r\n\r\nIn 1992, he motivates the concept of 'television and computer-based education” as means to reach large student numbers with only the best of teaching expertise and publishes an article on the concept in the SA Journal of Higher Education of 1993 (and later in 2003). The University of Pretoria subsequently approved a series of test projects on the concept with outreach to Mamelodi and Eerste Rust in 1993. In 1994, the University established a 'Unit for Telematic Education ' as a support section for multiple faculties at the University of Pretoria. In subsequent years, the concept of 'telematic education” subsequently becomes well established in academic circles in South Africa, grew in popularity, and is adopted by many universities and colleges throughout South Africa as a medium of enhancing education and training, as a method to reaching out to far out communities, and as a means to enhance study from the home environment.\r\n\r\nProfessor Snyman in subsequent years pursued research in semiconductor physics, semiconductor devices, microelectronics, and optoelectronics.\r\n\r\nIn 2000 he joined the TUT as a full professor. Here served for a period as head of the Department of Electronic Engineering. Here he makes contributions to solar energy development, microwave and optoelectronic device development, silicon photonics, as well as contributions to new mobile telecommunication systems and network planning in SA.\r\n\r\nCurrently, he teaches electronics and telecommunications at the TUT to audiences ranging from first-year students to Ph.D. level.\r\n\r\nFor his research in the field of 'Silicon Photonics” since 1990, he has published (as author and co-author) about thirty internationally reviewed articles in scientific journals, contributed to more than forty international conferences, about 25 South African provisional patents (as inventor and co-inventor), 8 PCT international patent applications until now. Of these, two USA patents applications, two European Patents, two Korean patents, and ten SA patents have been granted. A further 4 USA patents, 5 European patents, 3 Korean patents, 3 Chinese patents, and 3 Japanese patents are currently under consideration.\r\n\r\nRecently he has also published an extensive scholarly chapter in an internet open access book on 'Integrating Microphotonic Systems and MOEMS into standard Silicon CMOS Integrated circuitry”.\r\n\r\nFurthermore, Professor Snyman recently steered a new initiative at the TUT by introducing a 'Laboratory for Innovative Electronic Systems ' at the Department of Electrical Engineering. The model of this laboratory or center is to primarily combine outputs as achieved by high-level research with lower-level system development and entrepreneurship in a technical university environment. Students are allocated to projects at different levels with PhDs and Master students allocated to the generation of new knowledge and new technologies, while students at the diploma and Baccalaureus level are allocated to electronic systems development with a direct and a near application for application in industry or the commercial and public sectors in South Africa.\r\n\r\nProfessor Snyman received the WIRSAM Award of 1983 and the WIRSAM Award in 1985 in South Africa for best research papers by a young scientist at two international conferences on electron microscopy in South Africa. He subsequently received the SA Microelectronics Award for the best dissertation emanating from studies executed at a South African university in the field of Physics and Microelectronics in South Africa in 1987. In October of 2011, Professor Snyman received the prestigious Institutional Award for 'Innovator of the Year” for 2010 at the Tshwane University of Technology, South Africa. This award was based on the number of patents recognized and granted by local and international institutions as well as for his contributions concerning innovation at the TUT.",institutionString:null,institution:{name:"University of South Africa",country:{name:"South Africa"}}},{id:"317279",title:"Mr.",name:"Ali",middleName:"Usama",surname:"Syed",slug:"ali-syed",fullName:"Ali Syed",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/317279/images/16024_n.png",biography:"A creative, talented, and innovative young professional who is dedicated, well organized, and capable research fellow with two years of experience in graduate-level research, published in engineering journals and book, with related expertise in Bio-robotics, equally passionate about the aesthetics of the mechanical and electronic system, obtained expertise in the use of MS Office, MATLAB, SolidWorks, LabVIEW, Proteus, Fusion 360, having a grasp on python, C++ and assembly language, possess proven ability in acquiring research grants, previous appointments with social and educational societies with experience in administration, current affiliations with IEEE and Web of Science, a confident presenter at conferences and teacher in classrooms, able to explain complex information to audiences of all levels.",institutionString:null,institution:{name:"Air University",country:{name:"Pakistan"}}},{id:"75526",title:"Ph.D.",name:"Zihni Onur",middleName:null,surname:"Uygun",slug:"zihni-onur-uygun",fullName:"Zihni Onur Uygun",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/75526/images/12_n.jpg",biography:"My undergraduate education and my Master of Science educations at Ege University and at Çanakkale Onsekiz Mart University have given me a firm foundation in Biochemistry, Analytical Chemistry, Biosensors, Bioelectronics, Physical Chemistry and Medicine. After obtaining my degree as a MSc in analytical chemistry, I started working as a research assistant in Ege University Medical Faculty in 2014. In parallel, I enrolled to the MSc program at the Department of Medical Biochemistry at Ege University to gain deeper knowledge on medical and biochemical sciences as well as clinical chemistry in 2014. In my PhD I deeply researched on biosensors and bioelectronics and finished in 2020. Now I have eleven SCI-Expanded Index published papers, 6 international book chapters, referee assignments for different SCIE journals, one international patent pending, several international awards, projects and bursaries. In parallel to my research assistant position at Ege University Medical Faculty, Department of Medical Biochemistry, in April 2016, I also founded a Start-Up Company (Denosens Biotechnology LTD) by the support of The Scientific and Technological Research Council of Turkey. Currently, I am also working as a CEO in Denosens Biotechnology. The main purposes of the company, which carries out R&D as a research center, are to develop new generation biosensors and sensors for both point-of-care diagnostics; such as glucose, lactate, cholesterol and cancer biomarker detections. My specific experimental and instrumental skills are Biochemistry, Biosensor, Analytical Chemistry, Electrochemistry, Mobile phone based point-of-care diagnostic device, POCTs and Patient interface designs, HPLC, Tandem Mass Spectrometry, Spectrophotometry, ELISA.",institutionString:null,institution:{name:"Ege University",country:{name:"Turkey"}}},{id:"267434",title:"Dr.",name:"Rohit",middleName:null,surname:"Raja",slug:"rohit-raja",fullName:"Rohit Raja",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/267434/images/system/267434.jpg",biography:"Dr. Rohit Raja received Ph.D. in Computer Science and Engineering from Dr. CVRAMAN University in 2016. His main research interest includes Face recognition and Identification, Digital Image Processing, Signal Processing, and Networking. Presently he is working as Associate Professor in IT Department, Guru Ghasidas Vishwavidyalaya (A Central University), Bilaspur (CG), India. He has authored several Journal and Conference Papers. He has good Academics & Research experience in various areas of CSE and IT. He has filed and successfully published 27 Patents. He has received many time invitations to be a Guest at IEEE Conferences. He has published 100 research papers in various International/National Journals (including IEEE, Springer, etc.) and Proceedings of the reputed International/ National Conferences (including Springer and IEEE). He has been nominated to the board of editors/reviewers of many peer-reviewed and refereed Journals (including IEEE, Springer).",institutionString:"Guru Ghasidas Vishwavidyalaya",institution:{name:"Guru Ghasidas Vishwavidyalaya",country:{name:"India"}}},{id:"246502",title:"Dr.",name:"Jaya T.",middleName:"T",surname:"Varkey",slug:"jaya-t.-varkey",fullName:"Jaya T. Varkey",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/246502/images/11160_n.jpg",biography:"Jaya T. Varkey, PhD, graduated with a degree in Chemistry from Cochin University of Science and Technology, Kerala, India. She obtained a PhD in Chemistry from the School of Chemical Sciences, Mahatma Gandhi University, Kerala, India, and completed a post-doctoral fellowship at the University of Minnesota, USA. She is a research guide at Mahatma Gandhi University and Associate Professor in Chemistry, St. Teresa’s College, Kochi, Kerala, India.\nDr. Varkey received a National Young Scientist award from the Indian Science Congress (1995), a UGC Research award (2016–2018), an Indian National Science Academy (INSA) Visiting Scientist award (2018–2019), and a Best Innovative Faculty award from the All India Association for Christian Higher Education (AIACHE) (2019). She Hashas received the Sr. Mary Cecil prize for best research paper three times. She was also awarded a start-up to develop a tea bag water filter. \nDr. Varkey has published two international books and twenty-seven international journal publications. She is an editorial board member for five international journals.",institutionString:"St. Teresa’s College",institution:null},{id:"250668",title:"Dr.",name:"Ali",middleName:null,surname:"Nabipour Chakoli",slug:"ali-nabipour-chakoli",fullName:"Ali Nabipour Chakoli",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/250668/images/system/250668.jpg",biography:"Academic Qualification:\r\n•\tPhD in Materials Physics and Chemistry, From: Sep. 2006, to: Sep. 2010, School of Materials Science and Engineering, Harbin Institute of Technology, Thesis: Structure and Shape Memory Effect of Functionalized MWCNTs/poly (L-lactide-co-ε-caprolactone) Nanocomposites. Supervisor: Prof. Wei Cai,\r\n•\tM.Sc in Applied Physics, From: 1996, to: 1998, Faculty of Physics & Nuclear Science, Amirkabir Uni. of Technology, Tehran, Iran, Thesis: Determination of Boron in Micro alloy Steels with solid state nuclear track detectors by neutron induced auto radiography, Supervisors: Dr. M. Hosseini Ashrafi and Dr. A. Hosseini.\r\n•\tB.Sc. in Applied Physics, From: 1991, to: 1996, Faculty of Physics & Nuclear Science, Amirkabir Uni. of Technology, Tehran, Iran, Thesis: Design of shielding for Am-Be neutron sources for In Vivo neutron activation analysis, Supervisor: Dr. M. Hosseini Ashrafi.\r\n\r\nResearch Experiences:\r\n1.\tNanomaterials, Carbon Nanotubes, Graphene: Synthesis, Functionalization and Characterization,\r\n2.\tMWCNTs/Polymer Composites: Fabrication and Characterization, \r\n3.\tShape Memory Polymers, Biodegradable Polymers, ORC, Collagen,\r\n4.\tMaterials Analysis and Characterizations: TEM, SEM, XPS, FT-IR, Raman, DSC, DMA, TGA, XRD, GPC, Fluoroscopy, \r\n5.\tInteraction of Radiation with Mater, Nuclear Safety and Security, NDT(RT),\r\n6.\tRadiation Detectors, Calibration (SSDL),\r\n7.\tCompleted IAEA e-learning Courses:\r\nNuclear Security (15 Modules),\r\nNuclear Safety:\r\nTSA 2: Regulatory Protection in Occupational Exposure,\r\nTips & Tricks: Radiation Protection in Radiography,\r\nSafety and Quality in Radiotherapy,\r\nCourse on Sealed Radioactive Sources,\r\nCourse on Fundamentals of Environmental Remediation,\r\nCourse on Planning for Environmental Remediation,\r\nKnowledge Management Orientation Course,\r\nFood Irradiation - Technology, Applications and Good Practices,\r\nEmployment:\r\nFrom 2010 to now: Academic staff, Nuclear Science and Technology Research Institute, Kargar Shomali, Tehran, Iran, P.O. Box: 14395-836.\r\nFrom 1997 to 2006: Expert of Materials Analysis and Characterization. Research Center of Agriculture and Medicine. Rajaeeshahr, Karaj, Iran, P. O. Box: 31585-498.",institutionString:"Atomic Energy Organization of Iran",institution:{name:"Atomic Energy Organization of Iran",country:{name:"Iran"}}},{id:"248279",title:"Dr.",name:"Monika",middleName:"Elzbieta",surname:"Machoy",slug:"monika-machoy",fullName:"Monika Machoy",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/248279/images/system/248279.jpeg",biography:"Monika Elżbieta Machoy, MD, graduated with distinction from the Faculty of Medicine and Dentistry at the Pomeranian Medical University in 2009, defended her PhD thesis with summa cum laude in 2016 and is currently employed as a researcher at the Department of Orthodontics of the Pomeranian Medical University. She expanded her professional knowledge during a one-year scholarship program at the Ernst Moritz Arndt University in Greifswald, Germany and during a three-year internship at the Technical University in Dresden, Germany. She has been a speaker at numerous orthodontic conferences, among others, American Association of Orthodontics, European Orthodontic Symposium and numerous conferences of the Polish Orthodontic Society. She conducts research focusing on the effect of orthodontic treatment on dental and periodontal tissues and the causes of pain in orthodontic patients.",institutionString:"Pomeranian Medical University",institution:{name:"Pomeranian Medical University",country:{name:"Poland"}}},{id:"252743",title:"Prof.",name:"Aswini",middleName:"Kumar",surname:"Kar",slug:"aswini-kar",fullName:"Aswini Kar",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/252743/images/10381_n.jpg",biography:"uploaded in cv",institutionString:null,institution:{name:"KIIT University",country:{name:"India"}}},{id:"204256",title:"Dr.",name:"Anil",middleName:"Kumar",surname:"Kumar Sahu",slug:"anil-kumar-sahu",fullName:"Anil Kumar Sahu",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/204256/images/14201_n.jpg",biography:"I have nearly 11 years of research and teaching experience. I have done my master degree from University Institute of Pharmacy, Pt. Ravi Shankar Shukla University, Raipur, Chhattisgarh India. I have published 16 review and research articles in international and national journals and published 4 chapters in IntechOpen, the world’s leading publisher of Open access books. I have presented many papers at national and international conferences. I have received research award from Indian Drug Manufacturers Association in year 2015. My research interest extends from novel lymphatic drug delivery systems, oral delivery system for herbal bioactive to formulation optimization.",institutionString:null,institution:{name:"Chhattisgarh Swami Vivekanand Technical University",country:{name:"India"}}},{id:"253468",title:"Dr.",name:"Mariusz",middleName:null,surname:"Marzec",slug:"mariusz-marzec",fullName:"Mariusz Marzec",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/253468/images/system/253468.png",biography:"An assistant professor at Department of Biomedical Computer Systems, at Institute of Computer Science, Silesian University in Katowice. Scientific interests: computer analysis and processing of images, biomedical images, databases and programming languages. He is an author and co-author of scientific publications covering analysis and processing of biomedical images and development of database systems.",institutionString:"University of Silesia",institution:{name:"University of Silesia",country:{name:"Poland"}}},{id:"212432",title:"Prof.",name:"Hadi",middleName:null,surname:"Mohammadi",slug:"hadi-mohammadi",fullName:"Hadi Mohammadi",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/212432/images/system/212432.jpeg",biography:"Dr. Hadi Mohammadi is a biomedical engineer with hands-on experience in the design and development of many engineering structures and medical devices through various projects that he has been involved in over the past twenty years. Dr. Mohammadi received his BSc. and MSc. degrees in Mechanical Engineering from Sharif University of Technology, Tehran, Iran, and his PhD. degree in Biomedical Engineering (biomaterials) from the University of Western Ontario. He was a postdoctoral trainee for almost four years at University of Calgary and Harvard Medical School. He is an industry innovator having created the technology to produce lifelike synthetic platforms that can be used for the simulation of almost all cardiovascular reconstructive surgeries. He’s been heavily involved in the design and development of cardiovascular devices and technology for the past 10 years. He is currently an Assistant Professor with the University of British Colombia, Canada.",institutionString:"University of British Columbia",institution:{name:"University of British Columbia",country:{name:"Canada"}}},{id:"254463",title:"Prof.",name:"Haisheng",middleName:null,surname:"Yang",slug:"haisheng-yang",fullName:"Haisheng Yang",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/254463/images/system/254463.jpeg",biography:"Haisheng Yang, Ph.D., Professor and Director of the Department of Biomedical Engineering, College of Life Science and Bioengineering, Beijing University of Technology. He received his Ph.D. degree in Mechanics/Biomechanics from Harbin Institute of Technology (jointly with University of California, Berkeley). Afterwards, he worked as a Postdoctoral Research Associate in the Purdue Musculoskeletal Biology and Mechanics Lab at the Department of Basic Medical Sciences, Purdue University, USA. He also conducted research in the Research Centre of Shriners Hospitals for Children-Canada at McGill University, Canada. Dr. Yang has over 10 years research experience in orthopaedic biomechanics and mechanobiology of bone adaptation and regeneration. He earned an award from Beijing Overseas Talents Aggregation program in 2017 and serves as Beijing Distinguished Professor.",institutionString:null,institution:{name:"Beijing University of Technology",country:{name:"China"}}},{id:"89721",title:"Dr.",name:"Mehmet",middleName:"Cuneyt",surname:"Ozmen",slug:"mehmet-ozmen",fullName:"Mehmet Ozmen",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/89721/images/7289_n.jpg",biography:null,institutionString:null,institution:{name:"Gazi University",country:{name:"Turkey"}}},{id:"265335",title:"Mr.",name:"Stefan",middleName:"Radnev",surname:"Stefanov",slug:"stefan-stefanov",fullName:"Stefan Stefanov",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/265335/images/7562_n.jpg",biography:null,institutionString:null,institution:{name:"Medical University Plovdiv",country:{name:"Bulgaria"}}},{id:"242893",title:"Ph.D. Student",name:"Joaquim",middleName:null,surname:"De Moura",slug:"joaquim-de-moura",fullName:"Joaquim De Moura",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/242893/images/7133_n.jpg",biography:"Joaquim de Moura received his degree in Computer Engineering in 2014 from the University of A Coruña (Spain). In 2016, he received his M.Sc degree in Computer Engineering from the same university. He is currently pursuing his Ph.D degree in Computer Science in a collaborative project between ophthalmology centers in Galicia and the University of A Coruña. His research interests include computer vision, machine learning algorithms and analysis and medical imaging processing of various kinds.",institutionString:null,institution:{name:"University of A Coruña",country:{name:"Spain"}}},{id:"294334",title:"B.Sc.",name:"Marc",middleName:null,surname:"Bruggeman",slug:"marc-bruggeman",fullName:"Marc Bruggeman",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/294334/images/8242_n.jpg",biography:"Chemical engineer graduate, with a passion for material science and specific interest in polymers - their near infinite applications intrigue me. \n\nI plan to continue my scientific career in the field of polymeric biomaterials as I am fascinated by intelligent, bioactive and biomimetic materials for use in both consumer and medical applications.",institutionString:null,institution:null},{id:"255757",title:"Dr.",name:"Igor",middleName:"Victorovich",surname:"Lakhno",slug:"igor-lakhno",fullName:"Igor Lakhno",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/255757/images/system/255757.jpg",biography:"Igor Victorovich Lakhno was born in 1971 in Kharkiv (Ukraine). \nMD – 1994, Kharkiv National Medical Univesity.\nOb&Gyn; – 1997, master courses in Kharkiv Medical Academy of Postgraduate Education.\nPh.D. – 1999, Kharkiv National Medical Univesity.\nDSC – 2019, PL Shupik National Academy of Postgraduate Education \nProfessor – 2021, Department of Obstetrics and Gynecology of VN Karazin Kharkiv National University\nHead of Department – 2021, Department of Perinatology, Obstetrics and gynecology of Kharkiv Medical Academy of Postgraduate Education\nIgor Lakhno has been graduated from international training courses on reproductive medicine and family planning held at Debrecen University (Hungary) in 1997. Since 1998 Lakhno Igor has worked as an associate professor in the department of obstetrics and gynecology of VN Karazin National University and an associate professor of the perinatology, obstetrics, and gynecology department of Kharkiv Medical Academy of Postgraduate Education. Since June 2019 he’s been a professor in the department of obstetrics and gynecology of VN Karazin National University and a professor of the perinatology, obstetrics, and gynecology department. He’s affiliated with Kharkiv Medical Academy of Postgraduate Education as a Head of Department from November 2021. Igor Lakhno has participated in several international projects on fetal non-invasive electrocardiography (with Dr. J. A. Behar (Technion), Prof. D. Hoyer (Jena University), and José Alejandro Díaz Méndez (National Institute of Astrophysics, Optics, and Electronics, Mexico). He’s an author of about 200 printed works and there are 31 of them in Scopus or Web of Science databases. Igor Lakhno is a member of the Editorial Board of Reproductive Health of Woman, Emergency Medicine, and Technology Transfer Innovative Solutions in Medicine (Estonia). He is a medical Editor of “Z turbotoyu pro zhinku”. Igor Lakhno is a reviewer of the Journal of Obstetrics and Gynaecology (Taylor and Francis), British Journal of Obstetrics and Gynecology (Wiley), Informatics in Medicine Unlocked (Elsevier), The Journal of Obstetrics and Gynecology Research (Wiley), Endocrine, Metabolic & Immune Disorders-Drug Targets (Bentham Open), The Open Biomedical Engineering Journal (Bentham Open), etc. He’s defended a dissertation for a DSc degree “Pre-eclampsia: prediction, prevention, and treatment”. Three years ago Igor Lakhno has participated in a training course on innovative technologies in medical education at Lublin Medical University (Poland). Lakhno Igor has participated as a speaker in several international conferences and congresses (International Conference on Biological Oscillations April 10th-14th 2016, Lancaster, UK, The 9th conference of the European Study Group on Cardiovascular Oscillations). His main scientific interests: are obstetrics, women’s health, fetal medicine, and cardiovascular medicine. \nIgor Lakhno is a consultant at Kharkiv municipal perinatal center. He’s graduated from training courses on endoscopy in gynecology. He has 28 years of practical experience in the field.",institutionString:null,institution:null},{id:"244950",title:"Dr.",name:"Salvatore",middleName:null,surname:"Di Lauro",slug:"salvatore-di-lauro",fullName:"Salvatore Di Lauro",position:null,profilePictureURL:"https://intech-files.s3.amazonaws.com/0030O00002bSF1HQAW/ProfilePicture%202021-12-20%2014%3A54%3A14.482",biography:"Name:\n\tSALVATORE DI LAURO\nAddress:\n\tHospital Clínico Universitario Valladolid\nAvda Ramón y Cajal 3\n47005, Valladolid\nSpain\nPhone number: \nFax\nE-mail:\n\t+34 983420000 ext 292\n+34 983420084\nsadilauro@live.it\nDate and place of Birth:\nID Number\nMedical Licence \nLanguages\t09-05-1985. Villaricca (Italy)\n\nY1281863H\n474707061\nItalian (native language)\nSpanish (read, written, spoken)\nEnglish (read, written, spoken)\nPortuguese (read, spoken)\nFrench (read)\n\t\t\nCurrent position (title and company)\tDate (Year)\nVitreo-Retinal consultant in ophthalmology. Hospital Clinico Universitario Valladolid. Sacyl. National Health System.\nVitreo-Retinal consultant in ophthalmology. Instituto Oftalmologico Recoletas. Red Hospitalaria Recoletas. Private practise.\t2017-today\n\n2019-today\n\t\n\t\nEducation (High school, university and postgraduate training > 3 months)\tDate (Year)\nDegree in Medicine and Surgery. University of Neaples 'Federico II”\nResident in Opthalmology. Hospital Clinico Universitario Valladolid\nMaster in Vitreo-Retina. IOBA. University of Valladolid\nFellow of the European Board of Ophthalmology. Paris\nMaster in Research in Ophthalmology. University of Valladolid\t2003-2009\n2012-2016\n2016-2017\n2016\n2012-2013\n\t\nEmployments (company and positions)\tDate (Year)\nResident in Ophthalmology. Hospital Clinico Universitario Valladolid. Sacyl.\nFellow in Vitreo-Retina. IOBA. University of Valladolid\nVitreo-Retinal consultant in ophthalmology. Hospital Clinico Universitario Valladolid. Sacyl. National Health System.\nVitreo-Retinal consultant in ophthalmology. Instituto Oftalmologico Recoletas. Red Hospitalaria Recoletas. \n\t2012-2016\n2016-2017\n2017-today\n\n2019-Today\n\n\n\t\nClinical Research Experience (tasks and role)\tDate (Year)\nAssociated investigator\n\n' FIS PI20/00740: DESARROLLO DE UNA CALCULADORA DE RIESGO DE\nAPARICION DE RETINOPATIA DIABETICA BASADA EN TECNICAS DE IMAGEN MULTIMODAL EN PACIENTES DIABETICOS TIPO 1. Grant by: Ministerio de Ciencia e Innovacion \n\n' (BIO/VA23/14) Estudio clínico multicéntrico y prospectivo para validar dos\nbiomarcadores ubicados en los genes p53 y MDM2 en la predicción de los resultados funcionales de la cirugía del desprendimiento de retina regmatógeno. Grant by: Gerencia Regional de Salud de la Junta de Castilla y León.\n' Estudio multicéntrico, aleatorizado, con enmascaramiento doble, en 2 grupos\nparalelos y de 52 semanas de duración para comparar la eficacia, seguridad e inmunogenicidad de SOK583A1 respecto a Eylea® en pacientes con degeneración macular neovascular asociada a la edad' (CSOK583A12301; N.EUDRA: 2019-004838-41; FASE III). Grant by Hexal AG\n\n' Estudio de fase III, aleatorizado, doble ciego, con grupos paralelos, multicéntrico para comparar la eficacia y la seguridad de QL1205 frente a Lucentis® en pacientes con degeneración macular neovascular asociada a la edad. (EUDRACT: 2018-004486-13). Grant by Qilu Pharmaceutical Co\n\n' Estudio NEUTON: Ensayo clinico en fase IV para evaluar la eficacia de aflibercept en pacientes Naive con Edema MacUlar secundario a Oclusion de Vena CenTral de la Retina (OVCR) en regimen de tratamientO iNdividualizado Treat and Extend (TAE)”, (2014-000975-21). Grant by Fundacion Retinaplus\n\n' Evaluación de la seguridad y bioactividad de anillos de tensión capsular en conejo. Proyecto Procusens. Grant by AJL, S.A.\n\n'Estudio epidemiológico, prospectivo, multicéntrico y abierto\\npara valorar la frecuencia de la conjuntivitis adenovírica diagnosticada mediante el test AdenoPlus®\\nTest en pacientes enfermos de conjuntivitis aguda”\\n. National, multicenter study. Grant by: NICOX.\n\nEuropean multicentric trial: 'Evaluation of clinical outcomes following the use of Systane Hydration in patients with dry eye”. Study Phase 4. Grant by: Alcon Labs'\n\nVLPs Injection and Activation in a Rabbit Model of Uveal Melanoma. Grant by Aura Bioscience\n\nUpdating and characterization of a rabbit model of uveal melanoma. Grant by Aura Bioscience\n\nEnsayo clínico en fase IV para evaluar las variantes genéticas de la vía del VEGF como biomarcadores de eficacia del tratamiento con aflibercept en pacientes con degeneración macular asociada a la edad (DMAE) neovascular. Estudio BIOIMAGE. IMO-AFLI-2013-01\n\nEstudio In-Eye:Ensayo clínico en fase IV, abierto, aleatorizado, de 2 brazos,\nmulticçentrico y de 12 meses de duración, para evaluar la eficacia y seguridad de un régimen de PRN flexible individualizado de 'esperar y extender' versus un régimen PRN según criterios de estabilización mediante evaluaciones mensuales de inyecciones intravítreas de ranibizumab 0,5 mg en pacientes naive con neovascularización coriodea secunaria a la degeneración macular relacionada con la edad. CP: CRFB002AES03T\n\nTREND: Estudio Fase IIIb multicéntrico, randomizado, de 12 meses de\nseguimiento con evaluador de la agudeza visual enmascarado, para evaluar la eficacia y la seguridad de ranibizumab 0.5mg en un régimen de tratar y extender comparado con un régimen mensual, en pacientes con degeneración macular neovascular asociada a la edad. CP: CRFB002A2411 Código Eudra CT:\n2013-002626-23\n\n\n\nPublications\t\n\n2021\n\n\n\n\n2015\n\n\n\n\n2021\n\n\n\n\n\n2021\n\n\n\n\n2015\n\n\n\n\n2015\n\n\n2014\n\n\n\n\n2015-16\n\n\n\n2015\n\n\n2014\n\n\n2014\n\n\n\n\n2014\n\n\n\n\n\n\n\n2014\n\nJose Carlos Pastor; Jimena Rojas; Salvador Pastor-Idoate; Salvatore Di Lauro; Lucia Gonzalez-Buendia; Santiago Delgado-Tirado. Proliferative vitreoretinopathy: A new concept of disease pathogenesis and practical\nconsequences. Progress in Retinal and Eye Research. 51, pp. 125 - 155. 03/2016. DOI: 10.1016/j.preteyeres.2015.07.005\n\n\nLabrador-Velandia S; Alonso-Alonso ML; Di Lauro S; García-Gutierrez MT; Srivastava GK; Pastor JC; Fernandez-Bueno I. Mesenchymal stem cells provide paracrine neuroprotective resources that delay degeneration of co-cultured organotypic neuroretinal cultures.Experimental Eye Research. 185, 17/05/2019. DOI: 10.1016/j.exer.2019.05.011\n\nSalvatore Di Lauro; Maria Teresa Garcia Gutierrez; Ivan Fernandez Bueno. Quantification of pigment epithelium-derived factor (PEDF) in an ex vivo coculture of retinal pigment epithelium cells and neuroretina.\nJournal of Allbiosolution. 2019. ISSN 2605-3535\n\nSonia Labrador Velandia; Salvatore Di Lauro; Alonso-Alonso ML; Tabera Bartolomé S; Srivastava GK; Pastor JC; Fernandez-Bueno I. Biocompatibility of intravitreal injection of human mesenchymal stem cells in immunocompetent rabbits. Graefe's archive for clinical and experimental ophthalmology. 256 - 1, pp. 125 - 134. 01/2018. DOI: 10.1007/s00417-017-3842-3\n\n\nSalvatore Di Lauro, David Rodriguez-Crespo, Manuel J Gayoso, Maria T Garcia-Gutierrez, J Carlos Pastor, Girish K Srivastava, Ivan Fernandez-Bueno. A novel coculture model of porcine central neuroretina explants and retinal pigment epithelium cells. Molecular Vision. 2016 - 22, pp. 243 - 253. 01/2016.\n\nSalvatore Di Lauro. Classifications for Proliferative Vitreoretinopathy ({PVR}): An Analysis of Their Use in Publications over the Last 15 Years. Journal of Ophthalmology. 2016, pp. 1 - 6. 01/2016. DOI: 10.1155/2016/7807596\n\nSalvatore Di Lauro; Rosa Maria Coco; Rosa Maria Sanabria; Enrique Rodriguez de la Rua; Jose Carlos Pastor. Loss of Visual Acuity after Successful Surgery for Macula-On Rhegmatogenous Retinal Detachment in a Prospective Multicentre Study. Journal of Ophthalmology. 2015:821864, 2015. DOI: 10.1155/2015/821864\n\nIvan Fernandez-Bueno; Salvatore Di Lauro; Ivan Alvarez; Jose Carlos Lopez; Maria Teresa Garcia-Gutierrez; Itziar Fernandez; Eva Larra; Jose Carlos Pastor. Safety and Biocompatibility of a New High-Density Polyethylene-Based\nSpherical Integrated Porous Orbital Implant: An Experimental Study in Rabbits. Journal of Ophthalmology. 2015:904096, 2015. DOI: 10.1155/2015/904096\n\nPastor JC; Pastor-Idoate S; Rodríguez-Hernandez I; Rojas J; Fernandez I; Gonzalez-Buendia L; Di Lauro S; Gonzalez-Sarmiento R. Genetics of PVR and RD. Ophthalmologica. 232 - Suppl 1, pp. 28 - 29. 2014\n\nRodriguez-Crespo D; Di Lauro S; Singh AK; Garcia-Gutierrez MT; Garrosa M; Pastor JC; Fernandez-Bueno I; Srivastava GK. Triple-layered mixed co-culture model of RPE cells with neuroretina for evaluating the neuroprotective effects of adipose-MSCs. Cell Tissue Res. 358 - 3, pp. 705 - 716. 2014.\nDOI: 10.1007/s00441-014-1987-5\n\nCarlo De Werra; Salvatore Condurro; Salvatore Tramontano; Mario Perone; Ivana Donzelli; Salvatore Di Lauro; Massimo Di Giuseppe; Rosa Di Micco; Annalisa Pascariello; Antonio Pastore; Giorgio Diamantis; Giuseppe Galloro. Hydatid disease of the liver: thirty years of surgical experience.Chirurgia italiana. 59 - 5, pp. 611 - 636.\n(Italia): 2007. ISSN 0009-4773\n\nChapters in books\n\t\n' Salvador Pastor Idoate; Salvatore Di Lauro; Jose Carlos Pastor Jimeno. PVR: Pathogenesis, Histopathology and Classification. Proliferative Vitreoretinopathy with Small Gauge Vitrectomy. Springer, 2018. ISBN 978-3-319-78445-8\nDOI: 10.1007/978-3-319-78446-5_2. \n\n' Salvatore Di Lauro; Maria Isabel Lopez Galvez. Quistes vítreos en una mujer joven. Problemas diagnósticos en patología retinocoroidea. Sociedad Española de Retina-Vitreo. 2018.\n\n' Salvatore Di Lauro; Salvador Pastor Idoate; Jose Carlos Pastor Jimeno. iOCT in PVR management. OCT Applications in Opthalmology. pp. 1 - 8. INTECH, 2018. DOI: 10.5772/intechopen.78774.\n\n' Rosa Coco Martin; Salvatore Di Lauro; Salvador Pastor Idoate; Jose Carlos Pastor. amponadores, manipuladores y tinciones en la cirugía del traumatismo ocular.Trauma Ocular. Ponencia de la SEO 2018..\n\n' LOPEZ GALVEZ; DI LAURO; CRESPO. OCT angiografia y complicaciones retinianas de la diabetes. PONENCIA SEO 2021, CAPITULO 20. (España): 2021.\n\n' Múltiples desprendimientos neurosensoriales bilaterales en paciente joven. Enfermedades Degenerativas De Retina Y Coroides. SERV 04/2016. \n' González-Buendía L; Di Lauro S; Pastor-Idoate S; Pastor Jimeno JC. Vitreorretinopatía proliferante (VRP) e inflamación: LA INFLAMACIÓN in «INMUNOMODULADORES Y ANTIINFLAMATORIOS: MÁS ALLÁ DE LOS CORTICOIDES. RELACION DE PONENCIAS DE LA SOCIEDAD ESPAÑOLA DE OFTALMOLOGIA. 10/2014.",institutionString:null,institution:null},{id:"243698",title:"Dr.",name:"Xiaogang",middleName:null,surname:"Wang",slug:"xiaogang-wang",fullName:"Xiaogang Wang",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/243698/images/system/243698.png",biography:"Dr. Xiaogang Wang, a faculty member of Shanxi Eye Hospital specializing in the treatment of cataract and retinal disease and a tutor for postgraduate students of Shanxi Medical University, worked in the COOL Lab as an international visiting scholar under the supervision of Dr. David Huang and Yali Jia from October 2012 through November 2013. Dr. Wang earned an MD from Shanxi Medical University and a Ph.D. from Shanghai Jiao Tong University. Dr. Wang was awarded two research project grants focused on multimodal optical coherence tomography imaging and deep learning in cataract and retinal disease, from the National Natural Science Foundation of China. He has published around 30 peer-reviewed journal papers and four book chapters and co-edited one book.",institutionString:null,institution:null},{id:"7227",title:"Dr.",name:"Hiroaki",middleName:null,surname:"Matsui",slug:"hiroaki-matsui",fullName:"Hiroaki Matsui",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"University of Tokyo",country:{name:"Japan"}}},{id:"312999",title:"Dr.",name:"Bernard O.",middleName:null,surname:"Asimeng",slug:"bernard-o.-asimeng",fullName:"Bernard O. Asimeng",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"University of Ghana",country:{name:"Ghana"}}},{id:"318905",title:"Prof.",name:"Elvis",middleName:"Kwason",surname:"Tiburu",slug:"elvis-tiburu",fullName:"Elvis Tiburu",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"University of Ghana",country:{name:"Ghana"}}},{id:"336193",title:"Dr.",name:"Abdullah",middleName:null,surname:"Alamoudi",slug:"abdullah-alamoudi",fullName:"Abdullah Alamoudi",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Majmaah University",country:{name:"Saudi Arabia"}}},{id:"318657",title:"MSc.",name:"Isabell",middleName:null,surname:"Steuding",slug:"isabell-steuding",fullName:"Isabell Steuding",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Harz University of Applied Sciences",country:{name:"Germany"}}},{id:"318656",title:"BSc.",name:"Peter",middleName:null,surname:"Kußmann",slug:"peter-kussmann",fullName:"Peter Kußmann",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Harz University of Applied Sciences",country:{name:"Germany"}}}]}},subseries:{item:{id:"19",type:"subseries",title:"Animal Science",keywords:"Animal Science, Animal Biology, Wildlife Species, Domesticated Animals",scope:"The Animal Science topic welcomes research on captive and wildlife species, including domesticated animals. The research resented can consist of primary studies on various animal biology fields such as genetics, nutrition, behavior, welfare, and animal production, to name a few. Reviews on specialized areas of animal science are also welcome.",coverUrl:"https://cdn.intechopen.com/series_topics/covers/19.jpg",hasOnlineFirst:!0,hasPublishedBooks:!0,annualVolume:11415,editor:{id:"259298",title:"Dr.",name:"Edward",middleName:null,surname:"Narayan",slug:"edward-narayan",fullName:"Edward Narayan",profilePictureURL:"https://mts.intechopen.com/storage/users/259298/images/system/259298.jpeg",biography:"Dr. Edward Narayan graduated with Ph.D. degree in Biology from the University of the South Pacific and pioneered non-invasive reproductive and stress endocrinology tools for amphibians - the novel development and validation of non-invasive enzyme immunoassays for the evaluation of reproductive hormonal cycle and stress hormone responses to environmental stressors. \nDr. Narayan leads the Stress Lab (Comparative Physiology and Endocrinology) at the University of Queensland. A dynamic career research platform which is based on the thematic areas of comparative vertebrate physiology, stress endocrinology, reproductive endocrinology, animal health and welfare, and conservation biology. \nEdward has supervised 40 research students and published over 60 peer reviewed research.",institutionString:null,institution:{name:"University of Queensland",institutionURL:null,country:{name:"Australia"}}},editorTwo:null,editorThree:null,series:{id:"13",title:"Veterinary Medicine and Science",doi:"10.5772/intechopen.73681",issn:"2632-0517"},editorialBoard:[{id:"258334",title:"Dr.",name:"Carlos Eduardo",middleName:null,surname:"Fonseca-Alves",slug:"carlos-eduardo-fonseca-alves",fullName:"Carlos Eduardo Fonseca-Alves",profilePictureURL:"https://mts.intechopen.com/storage/users/258334/images/system/258334.jpg",institutionString:null,institution:{name:"Universidade Paulista",institutionURL:null,country:{name:"Brazil"}}},{id:"191123",title:"Dr.",name:"Juan 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