Betanin extraction from beetroot using different solvent systems.
\\n\\n
More than half of the publishers listed alongside IntechOpen (18 out of 30) are Social Science and Humanities publishers. IntechOpen is an exception to this as a leader in not only Open Access content but Open Access content across all scientific disciplines, including Physical Sciences, Engineering and Technology, Health Sciences, Life Science, and Social Sciences and Humanities.
\\n\\nOur breakdown of titles published demonstrates this with 47% PET, 31% HS, 18% LS, and 4% SSH books published.
\\n\\n“Even though ItechOpen has shown the potential of sci-tech books using an OA approach,” other publishers “have shown little interest in OA books.”
\\n\\nAdditionally, each book published by IntechOpen contains original content and research findings.
\\n\\nWe are honored to be among such prestigious publishers and we hope to continue to spearhead that growth in our quest to promote Open Access as a true pioneer in OA book publishing.
\\n\\n\\n\\n
\\n"}]',published:!0,mainMedia:null},components:[{type:"htmlEditorComponent",content:'
Simba Information has released its Open Access Book Publishing 2020 - 2024 report and has again identified IntechOpen as the world’s largest Open Access book publisher by title count.
\n\nSimba Information is a leading provider for market intelligence and forecasts in the media and publishing industry. The report, published every year, provides an overview and financial outlook for the global professional e-book publishing market.
\n\nIntechOpen, De Gruyter, and Frontiers are the largest OA book publishers by title count, with IntechOpen coming in at first place with 5,101 OA books published, a good 1,782 titles ahead of the nearest competitor.
\n\nSince the first Open Access Book Publishing report published in 2016, IntechOpen has held the top stop each year.
\n\n\n\nMore than half of the publishers listed alongside IntechOpen (18 out of 30) are Social Science and Humanities publishers. IntechOpen is an exception to this as a leader in not only Open Access content but Open Access content across all scientific disciplines, including Physical Sciences, Engineering and Technology, Health Sciences, Life Science, and Social Sciences and Humanities.
\n\nOur breakdown of titles published demonstrates this with 47% PET, 31% HS, 18% LS, and 4% SSH books published.
\n\n“Even though ItechOpen has shown the potential of sci-tech books using an OA approach,” other publishers “have shown little interest in OA books.”
\n\nAdditionally, each book published by IntechOpen contains original content and research findings.
\n\nWe are honored to be among such prestigious publishers and we hope to continue to spearhead that growth in our quest to promote Open Access as a true pioneer in OA book publishing.
\n\n\n\n
\n'}],latestNews:[{slug:"stanford-university-identifies-top-2-scientists-over-1-000-are-intechopen-authors-and-editors-20210122",title:"Stanford University Identifies Top 2% Scientists, Over 1,000 are IntechOpen Authors and Editors"},{slug:"intechopen-authors-included-in-the-highly-cited-researchers-list-for-2020-20210121",title:"IntechOpen Authors Included in the Highly Cited Researchers List for 2020"},{slug:"intechopen-maintains-position-as-the-world-s-largest-oa-book-publisher-20201218",title:"IntechOpen Maintains Position as the World’s Largest OA Book Publisher"},{slug:"all-intechopen-books-available-on-perlego-20201215",title:"All IntechOpen Books Available on Perlego"},{slug:"oiv-awards-recognizes-intechopen-s-editors-20201127",title:"OIV Awards Recognizes IntechOpen's Editors"},{slug:"intechopen-joins-crossref-s-initiative-for-open-abstracts-i4oa-to-boost-the-discovery-of-research-20201005",title:"IntechOpen joins Crossref's Initiative for Open Abstracts (I4OA) to Boost the Discovery of Research"},{slug:"intechopen-hits-milestone-5-000-open-access-books-published-20200908",title:"IntechOpen hits milestone: 5,000 Open Access books published!"},{slug:"intechopen-books-hosted-on-the-mathworks-book-program-20200819",title:"IntechOpen Books Hosted on the MathWorks Book Program"}]},book:{item:{type:"book",id:"6226",leadTitle:null,fullTitle:"MATLAB - Professional Applications in Power System",title:"MATLAB",subtitle:"Professional Applications in Power System",reviewType:"peer-reviewed",abstract:"Conventionally, the simulation of power engineering applications can be a challenge for both undergraduate and postgraduate students. 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Amyloidosis is defined as an insoluble protein fibril that is deposited, mainly, in the extracellular spaces of organs and tissues as a result of a sequence of changes in protein folding. Precursor proteins change their conformation that forms amyloid fibrils, then deposited amyloid induce organ damage with disease specific conditions.
To date, there are 27 types of amyloidosis known extracellular fibril proteins in human, and each amyloidosis is characterized amyloid protein precursor, systemic (S) or localized organ (L), and syndrome or involved tissues [1]. In the nomenclature, dialysis-related amyloidosis (DRA) is defined as β2-microglobulin-related (Aβ2M) amyloid which precursor protein is β2-microglobulin (β2-m). It is associated to dialysis, a kidney replacement therapy, and deposits in systemic (S), mainly joint tissues [1].
Long-term dialysis treatment for end-stage kidney disease often induces the Aβ2-m amyloid deposition in mainly osteoarticular tissues that induces various disorders, such as carpal tunnel syndrome (CTS), destructive spondyloarthropathy (DSA), and cystic bone lesions as well as in systemic organs such as heart and gastrointestinal tract when disease advances. Several biomolecules including β2-m as well as clinical risk factors are thought to relate with Aβ2M amyloidogenesis (Figure 1). Recent progress of dialysis therapy has improved survival of dialysis patients, however, older age and long-term dialysis treatment may increase the onset and acceleration of DRA. In this article, we described about DRA focused on pathogenesis, clinical manifestations and treatment, and showed DRA is still one of serious complications for patients undergoing long-term dialysis treatment.
Pathogenesis of dialysis-related amyloidosis (DRA). DRA is induced not only accumulation of β2-microglobulin but also several biomolecules and clinical risk factors.
β2-m is a polypeptide of 99 residues that has a molecular weight of 11.8 kDa. It forms the beta chain of the human leukocyte antigen (HLA) class I molecule and has a well-known β-sandwich structure that involves a 7-strand β-pleated structure stabilized with a single disulphide bond (Cys25-Cys80). β2-m changes the conformation under various in vivo or in vitro conditions, such as acidic pH [2], 2,2,2-trifluoroethanol (TFE) [3], sodium dodecyl sulfate (SDS) [4], lysophospholipids [5], non-esterified fatty acids [6], heating [7] and agitation [8]. Proper dose of those molecules induce conformational intermediate that is required for Aβ2M amyloid fibril formation/extension (see below section).
β2-m is a component of MHC class I molecules, which are present on all nucleated cells. Most β2-m is normally eliminated by the kidney via glomerular filtration and subsequent tubular catabolism with Megalin [9]. Thus, severe kidney damage induces the retention of β2-m in serum due to impaired excretion from the kidney.
Some clinical studies have attempted to identify the conformational intermediate form of circulating β2-m. Capillary electrophoresis reveals that patients undergoing hemodialysis due to end-stage kidney disease, but not healthy control subjects, have the conformational intermediate form of β2-m in serum [10]. The level of predialysis serum β2-m intermediate was 2.7±1.4 mg/L and native β2-m was 29.4±6.8 mg/L in 31 hemodialysis patients. Hemodialysis using a polymethylmethacrylate and online hemodiafiltration with a polysulfone membrane decreased the level of the native form, while any change in the intermediate form was variable [10]. These results suggest that intermediate β2-m is increased in hemodialysis patients and is difficult to remove with dialysis treatment. It may suggest that the intermediate form is immobilized in the extracellular space where Aβ2-m amyloid has a marked affinity for joint tissues (cartilage, capsule, and synovium). In addition, immunoaffinity–liquid chromatography–mass spectrometry analysis and immunoassay revealed the generation of lysine-58–cleaved and truncated β2-m (ΔK58-β2-m), which was found in serum from 20–40% HD patients but not in serum from control subjects [11]. However, this truncated form has not been demonstrated in the tissue containing Aβ2-m amyloid [12]. It is not certain whether the conformational intermediate or the truncated form of β2-m has a critical role of onset/progress of DRA, and future studies will be needed to understand the pathogenesis for Aβ2-m amyloid fibrils formation/extension.
A nucleation-dependent polymerization model explains the general mechanisms of amyloid fibril formation in vitro, in various types of amyloidosis [13-18]. This model consists of two phases, i.e., nucleation and extension phases. Nucleus formation requires a series of association steps of monomers, which are thermodynamically unfavorable, representing the rate-limiting step in amyloid fibril formation in vitro. Once the nucleus (n-mer) has been formed, further addition of monomers to the nucleus becomes thermodynamically favorable, resulting in rapid extension of amyloid fibrils according to a first-order kinetic model, i.e., via the consecutive association of precursor proteins onto the ends of existing fibrils [14, 17, 18].
β2-microglobulin-related (Aβ2M) amyloid fibril extension in vitro. Sub-micellar concentration of sodium dodecyl sulfate (SDS) induces Aβ2M amyloid fibrils extension at a neutral pH while micellar concentration induces amyloid fibril depolimerization in vitro. The amount of extended fibrils is measured by Thioflavin T (A), and observed by electron microscopy (B). Bar shows 250 nm.
In the mechanism of amyloidogenesis from β2-m, natively folded proteins, partial unfolding of protein is prerequisite to its assembly into amyloid fibrils [3, 13, 19, 20]. The extension of Aβ2M amyloid fibrils, as well as the formation of the fibrils from β2-m are greatly dependent on the pH of the reaction mixture, with the optimum pH around 2.0-3.0 [13, 14]. At pH 2.5, where the extension of Aβ2M amyloid fibrils is optimum, β2-m loses much of the secondary and tertiary structures observed at pH 7.5 [13, 19]. Aβ2M amyloid fibrils readily depolymerize into monomeric β2-m at a neutral pH [19], however, low concentration of TFE [3] and sub-micellar concentration of SDS [4] induced Aβ2M amyloid fibrils extension with changing conformation of β2-m monomer and inhibiting depolymerization of amyloid fibrils at a neutral pH in vitro (Figure 2). While TFE and SDS are organic compounds, several biomolecules could induce Aβ2M amyloidogenesis in vivo, such as apolipoprotein E, proteoglycans, glicosaminoglycans, type1 collagen, non-esterified fatty acid, and lysophospholipids. [4-6, 21, 22]. For example, some lysophospholipids, especially lysophosphatidic acid (LPA) induces both amyloid fibril formation and extension at a neutral pH [5]. The mechanism of amyloidogenesis is due to make β2-m monomer into partially unfolding the compact structure as well as stabilizing the extended fibrils in vitro. Clinically, plasma LPA concentration is higher in patients undergoing hemodialysis treatment as compare to healthy subjects [5]. It’s unclear the local concentration of lysophospholipids in the lesion that Aβ2M amyloid deposits, it may be reasonable to consider the reaction between β2-m and lysophospholipids that are increased in chronic kidney disease (CKD) undergoing dialysis treatment. Joint tissues that Aβ2M amyloid deposits at early stage in dialysis patients, contains many kinds of glycosaminoglycans and proteoglycans. Depolymerization of Aβ2M amyloid fibrils at a neutral pH in vitro was inhibited dose-dependently by the presence of some glycosaminoglycans (heparin, dermatan sulfate or heparin sulfate) or proteoglycans (biglycan, decorin or keratan sulfate proteoglycan) [21]. In addition, some glycosaminoglycans, especially heparin, enhanced the Aβ2M amyloid fibril extension induced by low concentration of TFE at a neutral pH [3]. These results suggest that some glycosaminoglycans and proteoglycans stabilize extended Aβ2M amyloid fibrils possibly by binding directly to the surface of the fibrils in vivo. Heparin is widely used for the hemodialysis treatment as an anticoagulant. Although no significant difference in the prevalence of Aβ2M amyloidosis was found between continuous ambulatory peritoneal dialysis patients and hemodialysis patients carefully matched for time on dialysis and age at the onset of dialysis [23], our study may suggest that heparin could exert a subtle effect for the development of Aβ2M amyloidosis under some clinical conditions.
Those molecules are picked up from the results of in vitro amyloid fibril formation, extension, and depolymerization, and further studies will be needed to analyze the histological relationship between those biomolecules and Aβ2M amyloid in the lesion.
It is not clearly understood the progress of bone disease after deposition of amyloid fibrils. Deposited amyloidosis induces compression that induced CTS and joint arthropathy. Also amyloid deposition induces local osteolysis that induced bone cysts and DSA. The progress through synovial inflammation and subsequent osteoclastogenesis and/or osteoclast activation through three possible pathways: (i) indirect action of inflammatory cytokines through the expression in osteoblasts of receptor activator of nuclear factor-κB ligand/osteoprotegerin ligand (RANKL/OPGL), (ii) direct action of inflammatory cytokines, and (iii) RANKL/OPGL expression in inflammatory cells.
Advanced CKD induces the serum level of β2-m to elevate due to the impaired metabolism and excretion in the kidney. The average serum concentration levels of β2-m in patients undergoing dialysis is significantly higher compared to those in normal subjects (25–45 vs. 1–2 mg/L) [10, 24-27]. It is clearly understood that the impairment of metabolism in the kidney is the main cause of fluid retention in HD patients; however, it is not clear whether the production of β2-m is increased with CKD and/or dialysis treatment. However, a study shows that the amount of β2-m on the surface of granulocytes, lymphocytes and monocytes in hemodialysis patients is higher than that in control subjects while mRNA expression of β2-m in blood cells is no significant difference between them [28]. This result shows the possibility that increased binding of β2-m to blood cells is one of major cause of retention of β2-m in dialysis patients. Thus continuous higher serum levels of β2-m could induced DRA after long-term dialysis treatment [24, 29].
Risk factors of DRA are (i) long-term dialysis treatment, (ii) initiation dialysis treatment in young age, (iii) hemodialysis treatment with low purity dialysate, (iv) use of low-flux dialysis membrane [30], while the pathogenesis of DRA with those risk factors is still incompletely understood. Recently it has trend to use high-flux dialysis membrane and high purity dialysate in hemodialysis treatment. However, progress of dialysis treatment as well as treatment for other diseases makes better survival of dialysis patients and older initiation of dialysis treatment. Thus long-term and old age dialysis patients increase, and DRA is still one of serious complications for patients undergoing dialysis treatment.
Serum level of β2-m, precursor protein of DRA, increase in dialysis patients and is believed most important for onset and progress of DRA. While cross sectional study shows no relation between onset of DRA and serum level of β2-m [24], DRA may be onset after accumulation of β2-m with long duration of dialysis treatment [29].
Long-term dialysis treatment for end-stage kidney disease often induces the Aβ2-m amyloid deposition in mainly osteoarticular tissues that induces various disorders, such as CTS, DSA, and cystic bone lesions as well as in rarely systemic organs such as heart [31] and gastrointestinal tract [32] when disease is advanced. CTS is induced by the deposition of Aβ2-m amyloid around synovium in carpal tunnel and the compression of median nerve. DSA is induced by the deposition of Aβ2-m amyloid and the defect of bone in spine. It is radiographically characterized by severe narrowing of the intervertebral disk space and erosions as well as cysts of the adjacent vertebral plates. DSA lesions are mostly detected in the highly mobile areas, such as C5–C7 and L3–L5 [33]. Cystic lesions occur in bones, such as carpal and femur that Aβ2-m amyloid deposition is found around them. Cystic lesions as well as mineral bone disorder associated with CKD increase the risk of bone fracture.
DRA, induce various osteo-articular disorders, is one of serious complications in patients undergoing long-term dialysis treatment even improvement of dialysis treatment such as dialysis membrane and dialysate [34, 35]. For example, we researched over a four year period 359 end-stage kidney disease patients undergoing dialysis treatment were admitted in our center for the treatment of their dialysis-related complications [34]. DSA was a major cause of hospital admission in the patients undergoing dialysis therapy for 20 years or more, and the rate increased along with the increasing duration of dialysis therapy. The incidence rate of histories of surgeries for osteoarticular disorders, related to DRA was 25.0, 66.0, and 77.8 % in 20-24 years, 25-29 years, and 30 years or more after the initiation of dialysis therapy, respectively (Figure 3). In the patients undergoing dialysis therapy for 30 years or more, the incidence rate of histories of surgeries for CTS, DSA, and joint arthropathy was 72.2%, 50.0%, and 22.2%, respectively that indicated they had analogous histories of surgeries for various osteoarticular disorders (Figure 4). These results indicate that the frequency and severity of osteoarticular disorders which may be caused by DRA accelerated with the increasing duration of dialysis therapy especially for the patients undergoing dialysis therapy for 30 years or more.
The incidence rate of histories of surgeries for osteo-articular disorders, related to dialysis-related amyloidosis (DRA). The rate is 25.0, 66.0, and 77.8 % in 20-24 years, 25-29 years, and 30 years or more after the initiation of dialysis therapy, respectively. In the patients undergoing dialysis therapy for 30 years or more, the rate for carpal tunnel syndrome (CTS), destructive spondyloarthropathy (DSA), and joint arthropathy was 72.2%, 50.0%, and 22.2%, respectively.
A case of long-term dialysis patients complicated with various osteo-articular disorders related to dialysis-related amyloidosis (DRA). A man had end-stage kidney disease due to chronic glomerulo nephritis and received hemodialysis treatment for 30 years. He had various DRA-related osteo-articlular disorders, such as carpal tunnel syndrome (CTS), joint artholopathy, and destructive spondyloartholopahty (DSA) that needed surgical treatment.
Next, we researched 102 patients undergoing dialysis treatment for 30 years or more in our related hospitals, and their complication of osteoarticular disorders. The age at initiation of dialysis therapy was 27.3 ± 7.7 years, and the duration of dialysis therapy was 32.3 ± 1.8 years. The surgery for CTS was done for 80 patients (76%) in 21.6 ± 5.5 years after the initiation of dialysis therapy. All of those patients received the surgeries more than 2 times, furthermore, some of them received more than 4 times for 30 years or more. The surgery for DSA was done for 17 patients (16%) in 27.1 ± 4.7 years after initiation of dialysis treatment. There is dissociation in the incidence of history of surgery for DSA between in our center [34] and in our related hospitals while that for CTS was similar. The main reason may be that our center is a university teaching hospital and severe patients are referred to our hospital. This is a major limitation, however, it should be remembered that one of the main causes for admission was DSA in the patients undergoing long-term dialysis therapy. Our results may suggest that DRA is one of most serious complications in the patients undergoing dialysis therapy for 30 years or more. Further study will be needed about the detail of DRA in the in long-term dialysis patients. In our studies, the main factor associated with osteoarticular disorders which may be caused by DRA was the duration of dialysis therapy despite the younger age at initiation of dialysis therapy [34]. Other risk factors, such as dialyzer and dialysate could not be considered because of the consistent improvements in the technologies from year to year. For example, long-term dialysis patients had used the low-flux dialyzer for several years since initiating therapy, but now use a high-flux dialyzer. Short-term patients however, have used the high-flux dialyzer since the initiation of dialysis therapy.
In summary of our clinical research, the frequency and seriousness of osteoarticular disorders which may be caused by DRA were accelerated with the duration of dialysis therapy, especially in cases treated for 30 years or more.
Main purpose of treatment for DRA is a) to prevent the deposition of Aβ2-m amyloid fibrils in the lesions, and b) to relieve symptoms induced by Aβ2-m amyloid deposition. Remove β2-m with dialysis treatment and suppression of systemic/local inflammation are beneficial to prevent the deposition of Aβ2-m amyloid fibrils. Practically, it should be used biocompatible high-flux dialysis membrane and high purity dialysate in hemodialysis treatment. In addition, hemofiltration, hemodiafiltration, and use of β2-m adsorption column have much effect to reduce β2-m and to improve symptoms [36]. Non-steroidal anti-inflammatory drugs or low dose of steroid sometimes show relief of symptoms induced by Aβ2-m amyloid deposition while use of steroid for long duration has risk to induce adverse effect, such as infection and osteoporosis. Surgical treatments are needed when Aβ2-m amyloid deposition induces severe osteoarticular symptoms.
The use of high-flux dialyzer membrane leads to a reduction in the serum level of β2-m as compared to using low-flux dialyzer membrane. In the HEMO Study [26], the predialysis serum β2-m level was lower in the high-flux membrane group than in the low-flux membrane group. In another study, switching of dialyzer from conventional to high-flux membrane reduced the predialysis serum β2-m level [37]. Clinically, Küchle et al [38]examined the effect of polysulfone high-flux dialysis membrane in hemodialysis treatment, and showed less onset of CTS, arthropathy and bone cysts as well as lower concentration of serum β2-m as compare to use of low-flux dialysis membrane. The reason why high-flux membrane produces a lower level of serum β2-m is not only that it promotes better clearance, but that it also increases the binding of β2-m to blood cells, such as granulocytes, lymphocytes and monocytes [28]. High purity dialysate with low endotoxin reduced serum β2-m, pentosidine, C-reactive protein, and interleukin-6 [39] that probably accelerates Aβ2-m amyloid deposition.
Hemodiafiltration has better clearance of middle size molecules than HD and is known to reduce the risk of progression of DRA. A recent multicenter prospective randomized study revealed that on line HDF showed greater efficiency than HD with low-flux membrane in reducing the basal level of β2-m [40].
A β2-m adsorption column has been developed as a way to directly eliminate serum β2-m. This adsorption column system is designed for direct hemoperfusion (Figure 5). Adsorption of β2-m by this column is a result both of hydrophobic and molecular size-dependent interactions between the ligand in the column and β2-m molecule. The effects of this column show the reduction rate; 60.0-78.9 %, the amount of adsorption; 157-300 mg, serum β2-m after treatment; 6.8-13.5 mg/L with single treatment [41-43].
A schematic representation of hemodialysis treatment with β2-microglobulin (β2-m) adsorption column. The β2-m adsorption column is placed in series with the dialyzer, with blood flowing through the column first.
According to a prospective multicenter study, a β2-m adsorption column that was placed in series with a polysulfone dialyzer increased serum β2-m reduction in patients undergoing hemodialysis as compare to hemodialysis treatment without β2-m adsorption column [42]. This study also showed improvements of DRA-related symptoms, such as joint pain and activity of daily living, and it may suggest that the column absorbs not only β2-m, but also other molecules related to inflammation. Furthermore, a clinical study showed shrink the size of bone cysts when they are checked by X-ray [43].
A significant inverse relationship is observed between residual renal function and serum β2-m level [44]. This suggests that peritoneal dialysis may keep lower serum levels of β2-m because of better maintenance of intrinsic renal function, but not peritoneal function, than hemodialysis. However the prevalence of histological DRA in peritoneal dialysis patients is not significantly different from that observed in a group of hemodialysis patients matched for age and dialysis duration [23]. End-stage kidney disease patients can do peritoneal dialysis only for 5-10 years, and it is difficult to discuss which treatment shows benefit to prevent DRA. A radical approach for DRA is kidney transplantation that reduces serum β2-m, improves symptoms related to DRA and inhibits the progression [45]. The effects of kidney transplantation on DRA probably due to not only recover of kidney function but also effect of immunosuppression therapy.
Use of steroid shows beneficial effect for the pain induced by DRA while surgical treatment will be needed for advanced CTS and DSA. However, DSA induces serious neurological symptoms and it is sometimes hard to relief them with surgery. For example, 95 of 865 patients undergoing dialysis treatment had surgeries for DSA, while rate of post-operative complications, such as infection and cardiac events, was much higher than those without DSA [46].
Chronic kidney disease-mineral and bone disorder (CKD-MBD) is a systemic disorder, which consists from abnormal levels of mineral-related bicochemistries, bone abnormalities, and soft tissue calcification [47]. Various types of bone abnormalities are observed in CKD patients, such as high-turnover bone disease and osteomalacia. DRA causes bone abnormalities in patients undergoing dialysis treatment. Bone cyst, joint arthropathy, and DSA are frequency and specifically found in patients especially undergoing long-term hemodialysis therapy [48]. It remains controversial whether DRA and related osteopathy should be included in CKD-MBD. However, DRA is at least closely involved with CKD-MBD, from the view point of preventing osteoarticular complications in dialysis patients (Figure 6).
Dialysis-related amyloidosis (DRA) in chronic kidney disease-mineral bone disorder (CKD-MBD). CKD-MBD as well as DRA contains various types of bone abnormalities. Furthermore, β2-microglobulin/DRA may be involved with cardiovascular disease, bone fracture, and mortality which are clinical outcomes of CKD-MBD. In the view point of bone abnormalities, DRA may be related strongly with CKD-MBD.
CKD- MBD contains various types of bone abnormalities, such as high-turnover bone disease, osteomalacia, and adyanmic bone disease. DRA, such as bone cyst, joint arthropathy and DSA, also causes bone abnormalities and is included in renal osteodystorphy. Recently, some groups reported the relation between serum levels of β2-m and atherosclerosis [49] or mortality [26], thus β2-m/DRA may be involved with cardiovascular disease, bone fracture, and mortality which are clinical outcomes of CKD-MBD. In the view point of bone abnormalities, DRA related osteopathy may enhance the serious bone disorder, such as bone fractures and DSA, in the presence of other bone abnormalities, such as high-turnover bone disease and osteomalacia. DRA is a serious complication in patients who are receiving long-term dialysis therapy and obviously seems more harmful than other osteodystrophy in terms of maintenance of their ADL and quality of life. Further studies will be needed for this assumption.
DRA is still one of major and serious complications in end-stage kidney disease patients undergoing long-term dialysis treatment. Several biomolecules that may relate to Aβ2M amyloidogenesis are raised from in vitro studies, and that will be needed to investigate the involvement in amyloid deposition in vivo. These findings will develop more beneficial prevention and treatment for DRA as well as improvement of dialysis treatment.
Vegetable beetroot (Beta vulgaris L.) has the notable scientific interest, because it is a rich source of nitrate (NO3−), a compound with advantageous cardiovascular health effects through the endogen production of nitric oxide (NO) [1, 2]. There are two classes of pigment in plants, i.e., betalains and anthocyanins. Beetroots are the chief sources of betalains which is a water-soluble nitrogen pigment with heterocyclic ring, which can be further subdivided into two classes depending on chemical structure: betaxanthins comprising indicaxanthin; vulgaxanthin I and II, accountable for orange-yellow coloring; and betacyanins, such as betanin, isobetanin, neobetanin, and prebetanin, accountable for red-violet coloring [3, 4]. The most abundant betacyanin is betanin (betanidin 5-O-β-
Chemical structure of betanin.
According to experimental studies, raw beetroot generally contains water (87.1%), carbohydrate (7.6%), protein (1.7%), fat (0.1%), and betanin (0.03–0.06%) [7]. In addition to natural food colorant property, betalain also exhibits antimicrobial, antiviral, and antioxidant activities [8]. Moreover, beetroot dye has nutrient value along with nontoxic nature; therefore, it even finds application in dyeing industry where the health aspect is a foremost criterion. Also, this natural dye extracted from beetroot is eco-friendly in nature and does not cause any environmental problems in contrast to the commercially available synthetic dyes [7].
The first report on the crystallization of betanin was communicated by two independent groups Schmidt and Schonleben [9] and Wyler and Dreiding [10]. These two groups employed an electrophoretic strategy for betanin purification. Wyler and Dreiding [11] recognized three products which were formed by the alkaline degradation of betanidin (Figure 2); these were 4-methylpyridine-2,6-dicarboxylic acid, formic acid, and S-cyclodopa(5,6-dihydroxy-2,3-dihydroindole 2-carboxylic acid). When these three products are placed in correct relationship with one another, they form the betanidin’s carbon skeleton and also the configuration of the second carbon [12].
Alkaline degradation of betanidin.
To identify the position of β-
Reactions that revealed the glucosyl residue position in betanin.
The yellow-colored compound, i.e., 5,6-di-O-methylneobetanidin trimethyl ester, converted into colorless compound, i.e., 5,6-di-O-methyl-2,3-dehydro-11,12-dihydro-betanidin trimethyl ester, by the palladium-catalyzed disproportionation reaction which confirmed the existence of vinylene connecting group (Figure 4) in the derivatives of neobetanidin [12].
Reaction which confirmed the presence of vinylene connecting group.
Further, betanin can also be interconverted into betanidin. To prepare betanidin, betanin is initially reacted with the excess of
The drawbacks of conventional approaches, such as time-consuming methods, safety risks with some hazardous solvent systems, contaminated product, and comparatively less yields, have increased the demand for the novel processing methods [16, 17]. To improve the betalain extraction, some of the pretreatment methods have been proposed, such as pulsed electric fields [18], gamma irradiation [8], and low-direct current electric fields [19]. But, such methods are quite expensive when compared to the solvent extraction techniques. Further, certain nonthermal techniques, like ultrasound (sonication) processing, and microwave-assisted extractions are also significantly productive in order to enhance the extraction yields of bioactive molecules with minimum degradation [17].
Neagu and Barbu [20] studied the betanin extraction from beetroot using different solvent systems by solid-liquid extraction technique (liquid/solid ratio is 5:1). Table 1 presents the different extraction solvents used in this study. Results revealed that the highest betanin content of about 20 mg/g of beetroot was obtained with the use of weak acid solution (i.e., V8, using 0.5% citric acid +0.1% ascorbic acid). Also, they extracted the considerable amount of betanins by using ascorbic acid added solutions. Thus, it is clear that the acidic medium influences positively during the low-temperature extraction process.
Variants | Solvents |
---|---|
V1 | Distilled water |
V2 | 1% citric acid |
V3 | 0.5% citric acid |
V4 | 0.2% citric acid |
V5 | 0.1% ascorbic acid |
V6 | 50% ethanol |
V7 | 20% ethanol |
V8 | 0.5% citric acid + 0.1% ascorbic acid |
V9 | 0.2% citric acid + 0.1% ascorbic acid |
V10 | 20% ethanol + 1% citric acid |
V11 | 20% ethanol + 0.5% citric acid |
Betanin extraction from beetroot using different solvent systems.
In case of ultrasound (sonication) processing, ultrasonic-assisted extraction approach requires the use of ultrasound (with 20–2000 kHz frequencies range) which generally increases the cell wall permeability and generates the cavitations. Because of cavitations, cell membrane breaks down, and internal materials (color and oil) come out [21].
On the other hand, microwave-assisted extractions are also contributed significantly to speed up the sample digestion and extraction of bioactive molecules from matrices. Here, the microwave energy has been utilized and employed in this extraction process. Moreover, this microwave energy induces molecular motion by the rotations of dipoles and migration of ions without varying the structure of the molecules provided the temperature of the system is not too high [21].
The biosynthetic pathways for the betanin molecule are depicted in Figure 5 [22]. Three enzymes such as 4,5-DOPA (dihydroxyphenylalanine)-extradiol-dioxygenase, tyrosinase, and betanidin-glucosyltransferase were involved in the biosynthesis of betalains in the cytoplasm [23]. From arogenic acid, the amino acid
Biosynthesis of betanin.
Finally, betanidin was formed by the formation of imine bond between cyclo-DOPA and betalamic acid which is then converted to betanin using betanidin-5-O-glucosyltransferase enzyme by connecting glucose unit of uridine diphosphate-glucose (UDP-G) to the hydroxyl group in position 5 [31]. But this reaction can be reversed in the presence of β-glucosidase [32]. Additionally, it is also concluded that enzyme cyclo-DOPA-5-O-glucosyltransferase catalyzes the transport of glucose molecule on cyclo-DOPA, by which the cyclo-DOPA-glucoside condense with betalamic acid to yield the betanin [33].
The chemical synthesis of betanidin is illustrated in Figure 6 [34]. For the synthesis of betanidin, 4-hydroxypyridine-2,6-dicarboxylic acid is used as the starting material which upon hydrogenation and followed by esterification yields all products in cis form. Further, Pfitzner-Moffatt oxidation reaction converted the secondary hydroxyl group to a ketone. In the next step, it is converted to semicarbazide using Horner-Wittig reagent. Then, the obtained semicarbazide is further hydrolyzed to give unsaturated ketone, which is converted to betalamic acid by Pfitzner-Moffatt oxidation. Reacting betalamic acid with
Chemical synthesis of betanidin.
The UV-vis spectra of beetroot extracts in different solvents (ethanol, methanol, and water) are depicted in Figure 7 [35]. Strong absorption band was observed at around 530 nm in the visible range for the red beet juice which was attributed to the betanin pigment. As the solvent changed from ethanol (532 nm) to water (542 nm) and to methanol (544 nm), absorption maximum shifted towards longer wavelengths. These results showed that methanolic and aqueous extracts mainly contain betanidin, whereas ethanolic extract mainly contains betanin. The intensity of absorption maxima for the aqueous and methanolic extracts is approximately equal, higher than the ethanolic extract.
The UV-vis spectra of beetroot extracts in different solvents (ethanol, methanol, and water).
Further, partially overlapped two absorption bands were observed in case of aqueous and methanolic extracts. For aqueous extract the second band has an absorption maximum at 515 nm and for aqueous extract at 509 nm. These bands are only observed in high concentrated extract, and it vanishes as the solution diluted. Hence, it can be attributed to the formation of supramolecular structure in the concentrated solutions.
Different characteristic absorption bands corresponding to the functional groups of betanins were observed, and its FT-IR spectrum is shown in Figure 8 [36]. The absorption band around 3359 cm−1 was ascribed to the ▬OH bond stretching vibration [37]; on the other hand, the absorption band around 1624 cm−1 was ascribed to the C〓N bond stretching vibration [38, 39]. The next absorption band located at 1378 cm−1 was ascribed to the C▬H bond extension stretching vibration, while the absorption band at 1243 cm−1 was ascribed to the C▬O bond of the carboxylic acid stretching vibration [38, 39]. Another absorption band centered at 1073 cm−1 was ascribed to the C▬O▬C linked symmetric stretching vibration [40], the absorption band at 945 cm−1 was ascribed to the C▬H bond deformation, and lastly the absorption band at 879 cm−1 was ascribed to the C▬COOH bond stretching vibrations [41].
FT-IR spectrum of betanin (scale range, 400–4000 cm−1).
The 1H and 13C NMR data of betanin was obtained by dissolving it in D2O, and its LC-1H NMR data was also obtained by dissolving it in acetonitrile (MeCN)/D2O/0.05% trifluoroacetic acid (TFA) using 500 MHz frequency at 25°C. Figure 9 represents the LC-1H NMR spectrum of betanin and followed by Table 2 which represents the 1H, 13C, and LC-1H NMR data of betanin [42].
LC-1H NMR spectrum of betanin.
Atom numbering | 1H NMR δ-[ppm], mult, J [Hz] | 13C NMR δ-[ppm], mult, J [Hz] | LC-1H NMR δ-[ppm], mult, J [Hz] |
---|---|---|---|
2 | 4.92, dd, 3.1, 10.3 | 65.0 | 5.23, dd, 2.0, 10.2 |
3a/b | 3.53, dd, 11.5, 16.9 3.10, dd, 4.3, 16.8 | 32.7 | 3.66, dd, 10.4, 16.7 3.31, dd, 2.0, 16.7 |
4 | 7.06, s | 113.9 | 7.13, s |
5 | — | 144.0 | — |
6 | — | 146.1 | — |
7 | 6.98, bs | 100.0 | 7.12, bs |
8 | — | 137.4 | — |
9 | — | 124.1 | — |
10 | — | 175.8 | — |
11 | 8.19, bs (d, 12.6)b | 144.4 | 8.38, bd, ≈11 |
12 | 5.84, bs (d, 12.6)b | 106.9 | 6.04, bd, ≈11 |
13 | — | —a | — |
14a/b | 3.20, bm 3.12, bm | 26.5 | 3.21, dd, 7.4, 17.2 3.41–3.59 (overlap) |
15 | 4.40, bt, 7.1 | 53.1 | Overlapped by D2O |
17 | — | —a | — |
18 | 6.22, bs | —a | 6.34, bs |
19 | — | —a | — |
20 | — | —a | — |
1′ | 4.98, d, 7.4 | 101.4 | 5.00, d, 6.9 |
2′ | 3.55 (overlap) | 75.7 | 3.41–3.59 (overlap) |
3′ | 3.55 (overlap) | 73.9 | 3.41–3.59 (overlap) |
4′ | 3.41 (overlap) | 69.3 | 3.41–3.59 (overlap) |
5′ | 3.52 (overlap) | 76.2 | 3.41–3.59 (overlap) |
6′a/b | 3.85, dd, 1.6, 12.3 3.70, dd, 5.3, 12.3 | 60.6 | 3.86, dd, 1.4, 12.6 3.70, dd, 5.5, 12.6 |
1H, 13C, and LC-1H NMR data of betanin.
Chemical shifts were not observable.bAfter acidification (TFA) to pH 2, s = singlet, d = doublet, dd = doublet of doublet, bs = broad singlet, bd = broad doublet, bt = broad triplet, bm = broad multiplet.
The mass spectrometry of betanin in the positive ionization mode exhibited a molecular ion (m/z 551, [M+H]+, 100%). Figure 10 represents the obtained mass spectrum of betanin molecule [43].
Mass spectrum of betanin.
The dynamic thermogravimetric (TG) analysis was conducted on fresh betanin and dried betanin (Figure 11a and b) [44]. Since fresh betanin contains water, immediate mass loss is noted in the temperature range of 40–100°C. Further, the degradation temperature of betanin dye is noted at the temperature of about 204°C. Furthermore, the dried sample has also exhibited similar behavior except the mass loss at the beginning (in the temperature range of 40–100°C).
(a) TG curves and (b) derivative TG curves of fresh and dried betanin.
pH: in the buffers of pH 2–9, betanin was stored at 4°C for 7 days and measured the visible spectra at both starting and end of this time span. No shifts in the absorption maxima were noted in between pH 4 and pH 7. A shift of about 2 nm towards a shorter wavelength with a decreased absorbance intensity was observed in case of the buffer which has pH less than 4. In the 575–650 nm region, the spectrum has slightly increased absorbance, and the solution color changed to red-violet from red. While in case of the solutions which has pH value of above 7, i.e., at pH 9, the absorption maximum moved to a longer wavelength region (544 nm) by decreasing the intensity. In the 575–650 nm and 400–450 nm wavelength regions, the absorbance increased to a considerable extent, and the solution color changed to violet from red. These results showed that between pH 3 and pH 7, storage had no effect on betanin solutions, and above and below of these pH values causes the considerable losses of betanin. Visible spectra of betanin compound at pH 2, 5, and 9 are illustrated in Figure 12 [12, 45].
Temperature: on heating the red color of betanin solutions starts to diminish, and finally it turns to brown color. The color loss was followed by the betanin assay, and the rate indicates that it follows first-order kinetics. The graph indicates that at 100°C, the degradation rate at pH 5 is still less than it is at pH 3 and pH 7. Further, the betanin compound is more stable between pH 4 and 5. However, betanin in beet juice is far more stable at pH 5, which reveals a protective effect by the constituents of juice. The rates of degradation for betanin molecule in a system at 100°C at pH 3, 5, and 7 are depicted in Figure 13 [12, 45].
Light: at 15°C and pH 7, the presence of light increased the rate of degradation by 15.6% and air (rather than nitrogen) by 14.6%. Both light and air together increased the rate by 28.6% [12, 42].
Visible spectra of betanin compound at pH 2, 5, and 9.
Rates of degradation for betanin molecule in a system at 100°C at pH 3, 5, and 7.
Pure betanin dye can compete with synthetic dyes in color depth shade properties and in color fastness properties. Guesmi et al. [43] studied the dyeing of betanin on modified acrylic fabrics and evaluated the effect of dye bath pH, salt concentration, dyeing time, and temperature on dyeing. In a dye bath having sodium chloride (0–15 g/L) and a dye of 30 mg/L concentration with the 40:1 liquor ratio, modified acrylic fabric was dyed using conventional heating.
Over the pH range 1–5, increase in pH increases the adsorption of betanin onto acrylic fabric. Color strength decreases as the pH increased above 5. Generally, amino functional groups of acrylic fibers get protonated as the pH value decreases. Thus, ion-ion forces induced with ionized carboxyl groups in betanin. Betanin may exist in cationized or on monoanion form in a strongly acidic environment which results in the lower depth of dyeing at pH less than 4, and also it is due to the betanin stability loss at low pH [43].
At pH 5 maximal color strength was observed, whereas at pH 4, little decrease in color strength was observed; this is attributed to the increased carboxyl groups in this range and to the high thermal stability of betanin molecule. The number of protonated terminal amino functional groups of fabric decreases at pH > 5, which causes the decreased ionic interaction between the carboxylate anion of the dye and acrylic fibers, thus lowering its dye ability. The structures of betanin molecule as a pH varied are depicted in Figure 14 [43].
Structures of betanin molecule as a pH varied [46].
Color strength decreases as the salt concentration increases, hence dyeing without salt addition is the best condition [43].
As the temperature of dyeing increases, the color strength increases up to 50°C, and further by increasing the dyeing temperature, the color strength decreased slowly which is attributed to the decrease in stability of dye at higher temperatures [43].
Color strength increases as the dyeing time increases up to 30 min; from 30 to 45 min, there is no change in color strength, and then it started to decrease as the time increases [43].
The fastness properties of betanin-dyed acrylic fabrics are shown in Table 3. The fastness properties of the dyed samples were examined according to ISO standard methods, the specific tests conducted for color fastness to rubbing is as per ISO 105-X12:1987, the color fastness to water is as per ISO 105-E01:1989, the color fastness to washing is as per ISO 105-C02:1989, and the color fastness to light is as per ISO 105-B02:1988 (carbon arc) [43]. It was noted that rubbing, washing, and water fastness of unmordanted acrylic fabrics exhibited significantly good property. But, the light fastness of unmordanted acrylic fabrics was found to be bad. However, light fastness was found to increase from rating 3* to 4* in premordanted fabrics using manganese sulfate and ferrous sulfate, and light fastness was increased from rating 3* to 5* by using cobalt sulfate. Nevertheless, the other mordants did not affect the light fastness of premordanted fabrics. It was found that for the improvements of color strength and light fastness, cobalt sulfate was established as the best mordant [43].
Fastness to dry rubbing | Fastness to wet rubbing | Fastness to washing | Fastness to water | Fastness to light | |
---|---|---|---|---|---|
None | 5* | 5* | 4*–5* | 4*–5* | 3* |
Manganese sulfate | 5* | 5* | 4* | 4* | 4* |
Ferrous sulfate | 4* | 4* | 3* | 3* | 3* |
Zinc sulfate | 5* | 5* | 4*–5* | 4*–5* | 4* |
Aluminum potassium sulfate | 4* | 4* | 3* | 3* | 3* |
Cobalt sulfate | 5* | 5* | 4*–5* | 4*–5* | 5* |
The fastness properties of betanin-dyed acrylic fabrics represented with the rating scale using star (*).
Similarly, Guesmi et al. [46] in the year 2013 studied the dyeing of wool fabric using betanin and chlorophyll-a as biomordant. In a dye bath having sodium chloride (0–5 g/L) and a dye with 40:1 liquor ratio, wool fabric was dyed using conventional heating. Results revealed that the increase in the concentration of biomordant increases the color strength values. They also investigated the effect of variables on the color of dyed fibers and noted that from pH 3.5 to pH 4.5, the color strength considerably increases, the color strength was found to be better without salt than with salt, and the color strength of dyed wool increases as the increase in temperature was up to 40°C and starts to decrease slowly till 50°C. Further increase in temperature, the color strength decreases in pronounced manner. According to the authors, color strength increases with the time span (up to 45 min) of dyeing, and then it starts to decrease because betanin losses thermal stability, and also it starts to escape from the fiber. Dye exhaustion was examined in both ultrasonic and conventional dyeing approaches. It was exhibited that in a shorter time span of dyeing, sonication increases the dye exhaustion from rating of 30% to rating of 60%. The fastness properties of dyed wool were studied against wet rubbing, light, washing, and dry rubbing. Unmordanted and mordanted samples have good fastness properties too.
Antioxidant activity of betanin in biological lipid domain has been exhibited in human macromolecules, like lipoproteins of low density, whole cells, and membranes [2]. Moreover, betanin has attracted researchers because of its anti-inflammatory activities and hepatic safety activities in whole human cells [47]. In cultured endothelia cells, this molecule regulates the redox-mediated signal transduction pathways which is required in responses during inflammation, and betanin also showed antiproliferative effects on tumor cell lines in human [48, 49, 50]. In both tumoral and healthy hepatic cell lines in the human body, betanin translocates the antioxidant response element (erythroid 2-related factor 2 (Nrf2)) from the place of cytosol to the place of nuclear domain, which regulates m-RNA and protein levels of antioxidant/detoxifying enzymes, which includes heme oxygenase-1, NAD(P)H quinone dehydrogenase-1, and glutathione S-transferase and, in these cells, bears anticarcinogenic and hepatoprotective effects [51]. Also, it exhibits antidiabetes properties by controlling the activities of liver markers enzymes [52, 53, 54].
Betanin is the oldest and most abundant red food colorant which has been established in the market, which is noted as E-162 in the European Union and in the United Sates; it is known as 73.40 in the twenty-first chapter of the Code of Federal Regulations (CFR) section of the Food and Drug Earth Administration [2, 5, 6].
Betanins are most commonly used for coloring of ice cream and powdered soft drink beverages. Additionally, betanin is used in some of the sugar confectionery, like sugar coatings, fruit or ice cream fillings, fondants, and sugar strands. At the final part of the processing, it can be added while preparing hot processed candies. Also, it is used in soups as well as bacon and tomato.
In this chapter, the first and second section covered the chemistry of betanin which contains reactions that revealed the glucosyl residue position and presence of vinylene connecting group in betanin. Further, the third section narrated the extraction techniques which mainly included the microwave- and ultrasonic-assisted extraction method. Furthermore, the fourth and fifth sections elucidated the biosynthesis of betanin molecule and chemical synthesis of betanidin molecule, respectively. In addition, different characterization techniques were also explicated in the sixth section which includes UV-Vis, FT-IR, 1H NMR, 13C NMR, LC-1H NMR, mass spectrum, and thermogravimetric analysis of betanin. Also, the factors effecting the stability of betanin were explained in the seventh section which covers the effect of pH, temperature, and light on the stability of betanin. Lastly, the applicability of betanin was taken into account in the eighth section which comprised of dyeing of acrylic fabric, dyeing of wool fabric, and medicinal and food colorant applications of betanin.
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