Browsing by Author "Pramod Kumar Srivastava"

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A molecular description of acid phosphatase
(2012) Asha Anand; Pramod Kumar Srivastava
Acid phosphatase is ubiquitous in distribution in various organisms. Although it catalyzes simple hydrolytic reactions, it is considered as an interesting enzyme in biological systems due to its involvement in different physiological activities. However, earlier reviews on acid phosphatase reveal some fragmentary information and do not give a holistic view on this enzyme. So, the present review summarizes studies on biochemical properties, structure, catalytic mechanism, and applications of acid phosphatase. Recent advancement of acid phosphatase in agricultural and clinical fields is emphasized where it is presented as potent agent for sustainable agricultural practices and diagnostic marker in bone metabolic disorders. Also, its significance in prostate cancer therapies as a therapeutic target has been discussed. At the end, current studies and prospects of immobilized acid phosphatase are included. © Springer Science+Business Media, LLC 2012.
A study of NADP+- linked isocitrate dehydrogenase from germinating mung bean (Vigna radiata)
(Society for Plant Biochemistry and Biotechnology, 2004) Pramod Kumar Srivastava; Kavita Pathak; Om Prakash
NADP+- linked isocitrate dehydrogenase has been purified to apparent homogeneity from 36 h germinated mung beans by ammonium sulphate fractionation, heat treatment, acid treatment, and DEAE - Cellulose column chromatography. The enzyme was purified to 150 fold with 15% recovery. The preparation showed single protein band on native PAGE and was free from bound nucleotides and coloured pigments (A280/A260 = 1.4). The molecular weight was found to be 141,000 and was made of four identical subunits (mol wt 36,000). Thermal inactivation at 50, 53, and 55°C revealed simple first order kinetics and t1/2 was found to be 38, 10, and 3 min, respectively. The enzyme exhibited absolute specificity for NADP+ and substrate. The Km for isocitrate and NADP+ was 28.57 μM and 70 μM, respectively. The enzyme appeared to be regulated by various metabolites of Krebs' cycle intermediates.
Allethrin-induced genotoxicity and oxidative stress in Swiss albino mice
(Elsevier B.V., 2012) Amit Kumar Srivastava; Pramod Kumar Srivastava; Abdulaziz A. Al-Khedhairy; Javed Musarrat; Yogeshwer Shukla
Allethrin (C19H26O3) is non-cyano-containing pyrethroid insecticide that is used extensively for controlling flies and mosquitoes. Apart from its neurotoxic effects in non-target species, allethrin is reported to be mutagenic in bacterial systems. In this study, we observed oxidative damage-mediated genotoxicity caused by allethrin in Swiss albino mice. The genotoxic potential of allethrin was evaluated using chromosome aberrations (CAs) and a micronuclei (MN) induction assay as genetic end-points. The oral intubation of allethrin (25 and 50mg/kg b.wt.) significantly induces CAs and MN in mouse bone marrow cells. The DNA-damaging potential of allethrin was estimated in mouse liver using the DNA alkaline unwinding assay (DAUA) and by measuring the levels of 8-hydroxy-2'-deoxy-guanosine (8-OH-dG). Furthermore, a dose-dependent increase in reactive oxygen species (ROS) generation and lipid peroxidation (LPO), with a concurrent decrease in superoxide dismutase (SOD) and catalase, confirm its pro-oxidant potential. The DNA-damaging potential of allethrin was found to be mediated through the modulation of p53, p21, GADD45α and MDM-2. These results confirm the genotoxic and the pro-oxidant potential of allethrin in Swiss albino mice. © 2012.
Association of single nucleotide polymorphisms in CYP1B1 and COMT genes with breast cancer susceptibility in Indian women
(Hindawi Limited, 2009) Sharawan Yadav; Naveen Kumar Singhal; Virendra Singh; Neeraj Rastogi; Pramod Kumar Srivastava; Mahendra Pratap Singh
Cytochrome P450 1B1 (CYP1B1) and catechol-$O$- methyltransferase (COMT) enzymes play critical roles in estrogen metabolism. Alterations in the catalytic activity of CYP1B1 and COMT enzymes have been found associated with altered breast cancer risk in postmenopausal women in many populations. The substitution of leucine (Leu) to valine (Val) at codon 432 increases the catalytic activity of CYP1B1, however, substitution of Val to methionine (Met) at codon 158 decreases the catalytic activity of COMT. The present study was performed to evaluate the associations of CYP1B1 Leu 432Val and/or COMT Val158Met polymorphisms with total, premenopausal and postmenopausal breast cancer risks in Indian women. COMT and CYP1B1 polymorphisms in controls and breast cancer patients were analyzed employing polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) followed by gel electrophoresis. Although CYP1B1 and COMT genotypes did not exhibit statistically significant association with breast cancer risks when analyzed individually, COMT wild type (Val158Val) in combination with CYP1B1 heterozygous variant (Leu432Val) [OR: 0.21; 95% CI (0.05-0.82), p value; 0.021] and COMT heterozygous variant (Val 158Met) in combination with CYP1B1 wild type (Leu432Leu) [OR: 0.29; 95% CI (0.08-0.96), p value; 0.042] showed significant protective association with premenopausal breast cancer risk. The results demonstrate that CYP1B1 wild type in combination with COMT heterozygous or their inverse combination offer protection against breast cancer in premenopausal Indian women. © 2009 - IOS Press and the authors. All rights reserved.
Changing epidemiology of community-acquired acute kidney injury in developing countries: Analysis of 2405 cases in 26 years from eastern India
(2013) Jai Prakash; Takhellambam Brojen Singh; Biplab Ghosh; Vinay Malhotra; Surendra Singh Rathore; Rubina Vohra; Rabindra Nath Mishra; Pramod Kumar Srivastava; Usha
BackgroundThe epidemiology of acute kidney injury (AKI) differs from country to country and varies from center to center within a country. Owing to the absence of a central registry, data on overall epidemiology of AKI are scanty from India.MethodsThis study aimed at describing changes in epidemiology of community-acquired AKI (CAAKI) over a time span of 26 years in two study periods, namely, 1983-95 and 1996-2008.ResultsWe studied 2405 (1375 male and 1030 female) cases of AKI in the age range 1-95 (mean: 40.32) years. The incidence of CAAKI in 1983-95 and 1996-2008 was 1.95 and 4.14 per 1000 admission, respectively (P < 0.01). Obstetrical AKI has decreased because of the declining number of post-abortal AKI. Surgical AKI decreased from 13.8% in 1983-95 to 9.17% in 1996-2008(P < 0.01). Malarial AKI increased significantly from 4.7% in the first half of the study to 17% in the later period (P < 0.01). Diarrhea-associated AKI had significantly decreased from 36.83% in 1983-95 to 19% in 1996-2008 (P < 0.01). Sepsis-related AKI had increased from 1.57% in 1983-95 to 11.43% in 1996-2008 (P < 0.01). Nephrotoxic AKI showed an increasing trend in recent years (P < 0.01) and mainly caused by rifampicin and NSAIDs. Liver disease-related AKI increased from 1.73% in 1983-95 to 3.17% in 1996-2008 (P < 0.01). Myeloma-associated acute renal failure (ARF) accounted for 1.25% of the total number of ARF cases in the period 1996-2008. HIV infection contributed to 1.65% of ARF of the total number of AKI cases in the second period (1996-2008). Incidence of renal cortical necrosis (RCN) decreased significantly from 5.8% in 1983-95 to 1.3% in 1996-2008 of the total number of ARF cases (P < 0.01). However, during the same period ARF due to acute tubular necrosis, acute glomerulonephritis and acute interstitial nephritis remained unchanged. The mortality rate from AKI decreased significantly from 20% in 1983-95 to 10.98% in 1996-2008 (P < 0.01).ConclusionsThe epidemiological characteristics of CAAKI have changed over the past three decades. There has been an increase in the overall incidence of ARF with the changing etiology of AKI in recent years. Incidences of obstetrical, surgical and diarrheal AKI have decreased significantly, whereas those of AKI associated with malaria, sepsis, nephrotoxic drugs and liver disease have increased. RCN has decreased significantly. In contrast to developed nations, community-acquired AKI is more common in developing countries. It often affects younger individuals and is caused by single and preventable diseases. © 2013 The Author.
Identification and characterization of cellular locus of limonin biotransforming enzyme in Pseudomonas putida
(2010) Jay Prakash Verma; Siddhartha Singh; Moushumi Ghosh; Pramod Kumar Srivastava
Limonin is a highly oxygenated triterpene derivative of class of limonoids which causes delayed bitterness in citrus. Pseudomonas putida (P. putida) is able to reduce 47% and 63% of limonin from juice serum extract and standard limonin, respectively. Biochemical studies with P. putida indicate that probably two metabolic pathways viz. 17-dehydrolimonoid and deoxylimonoid pathway exists in the test organism. Experimental results indicate that the enzyme limonoate dehydrogenase which is found to be localized in periplasmic space of P. putida plays a major role in conversion of limonin to 17-dehydrolimonoate A-ring lactone. Enzymatic studies have shown a 72% reduction in limonin. The experimental results show 9 folds reduction in limonin content in presence of NAD as cofactor. The molecular weight of one of its polypeptide is found to be 66 kDa. The optimum pH and temperature for enzyme activity is 8.5 and 30°C, respectively. © 2010 The Authors. Journal compilation © 2010 Institute of Food Science and Technology.
Immobilization and applications of glucose-6-phosphate dehydrogenase: A review
(2013) Pramod Kumar Srivastava; Siddhartha Singh
Immobilized enzymes have been used extensively in the fields of food industry, materials processing, textiles, detergents, biochemical and chemical industries, biotechnology, and pharmaceuticals. Studies on immobilization of glucose-6-phosphate dehydrogenase have been less extensive than those for other industrially applicable enzymes. Immobilization of glucose-6-phosphate dehydrogenase has been carried out for the formation of biosensors for the estimation of glucose, ATP, phosphate, and so on. The present review deals with the attempts made for immobilization of glucose-6-phosphate dehydrogenase and its applications for various purposes. © 2013 Taylor & Francis Group, LLC.
Immobilization of acid phosphatase from Vigna aconitifolia seeds on chitosan beads and its characterization
(Elsevier B.V., 2014) Pramod Kumar Srivastava; Asha Anand
Acid phosphatase isolated from Vigna aconitifolia seeds was immobilized onto glutaraldehyde activated chitosan beads by crosslinking method. Chitosan beads activated with 2% of glutaraldehyde have demonstrated maximum immobilization yield (~83%). The immobilized enzyme showed optimum activity at pH 7.0, while soluble form was maximally active in acidic range (pH 5.0). With respect to free form, immobilized acid phosphatase showed better activity in alkaline range. On the other side, immobilization does not affect the optimum temperature range i.e., both, soluble and immobilized acid phosphatase exhibited maximum activity at 60°C. The Km and Vmax values for the immobilized enzyme were calculated to be 0.37mM and 13.5U/mg. The immobilization on chitosan beads enhanced the shelf life of acid phosphatase. The immobilized enzyme retained its more than 50% hydrolytic activity for approximately two months. The immobilized acid phosphatase was reusable for more than 40 cycles of reaction. © 2013 Elsevier B.V.
In vivo efficacy and synergistic interaction of 16α-hydroxycleroda-3, 13 (14) Z-dien-15, 16-olide, a clerodane diterpene from Polyalthia longifolia against methicillin-resistant Staphylococcus aureus
(2013) Vivek Kumar Gupta; Surjeet Verma; Anirban Pal; Santosh Kumar Srivastava; Pramod Kumar Srivastava; Mahendra P. Darokar
The Staphylococcus aureus bacterium, a nosocomial pathogen often causing untreatable and lethal infection in patients, mutated to become resistant to all the first-line drugs. The present study details the potential of clerodane diterpene 16α-hydroxycleroda-3, 13 (14) Z-dien-15, 16-olide (CD) isolated from Polyalthia longifolia against methicillin-resistant S. aureus (MRSA) through in vitro and in vivo assays. Minimum inhibitory concentration (MIC) of CD exhibited significant anti-MRSA activity (15.625-31.25 mg/l) against reference strain and seven clinical isolates, while time kill assays at graded MICs indicated 2.78-9.59- and 2.9-6.18-fold reduction in growth of reference strain and clinical isolates of S. aureus, respectively. The combined effect of the CD and 7.5 % NaCl resulted in significant reduction in microbial count within 24 h, indicating the loss of the salt tolerance ability of S. aureus. Further, release of 260-nm absorbing material and flow cytometric analysis revealed an increased uptake of propidium iodide. These assays may indicate the membrane-damaging potential of CD. The molecule CD was found to interact synergistically with clinically used antibiotics (FICI ≤ 0.5) against all clinical isolates. In infected mice, CD significantly (P < 0.001) lowered the systemic microbial load in blood, liver, kidney, lung and spleen tissues and did not exhibit any significant toxicity at 100 mg/kg body weight. © 2013 Springer-Verlag Berlin Heidelberg.
Isolation and enzymatic properties of a nonspecific acid phosphatase from Vigna aconitifolia seeds
(Wiley-Blackwell Publishing Ltd, 2014) Asha Anand; Pramod Kumar Srivastava
Acid phosphatase (EC 3.1.3.2) from Vigna aconitifolia seeds was purified to apparent homogeneity by using ammonium sulfate fractionation and cation-exchange chromatography [carboxymethyl (CM) cellulose]. The enzyme was 228-fold purified with 14.6% recovery. Analytical gel filtration chromatography on Sephadex G-200 column showed that Mr of native enzyme was 58 kDa and denaturing PAGE demonstrated that it was made up of two subunits of 24 and 27 kDa. The enzyme showed its optimum activity at pH 5.0 and 60°C. It exhibited broad substrate specificity and showed a higher specificity constant for para-nitrophenyl phosphate, Na β-naphthyl phosphate, and adenosine monophosphate (AMP). Cu2+, Mo6+, Fe3+, phosphate, and fluoride ions were reported as strong inhibitors for the enzyme. Active site study for the enzyme demonstrated that tryptophan and aspartic acid may be important for the catalysis. © 2013 International Union of Biochemistry and Molecular Biology, Inc.
Mancozeb-induced genotoxicity and apoptosis in cultured human lymphocytes
(Elsevier Inc., 2012) Amit Kumar Srivastava; Wahid Ali; Richa Singh; Kulpreet Bhui; Shilpa Tyagi; Abdulaziz A. Al-Khedhairy; Pramod Kumar Srivastava; Javed Musarrat; Yogeshwer Shukla
Aims: Mancozeb is a dithiocarbamate fungicide known to be genotoxic and induces tumors in rodents at various sites. There is no report in the literature about its genotoxicity in humans. Here, we investigated the association between mancozeb exposure and induction of genotoxic and proapoptotic changes in cultured human lymphocytes (CHLs). Main methods: Lymphocytes were isolated from peripheral blood of healthy non-smoking donors. Induction of micronuclei and chromosomal aberrations was recorded both by conventional and flow cytometric methods. Annexin-V FITC was used for the differentiation of apoptotic and necrotic cells by flow cytometry. Key findings: Mancozeb exposure (0.5, 2 and 5 μg/ml) to CHLs leads to significant induction in the frequency of chromosomal aberrations (CAs) and micronuclei (MN), in a dose-dependent manner. Concomitantly, pro-oxidant potential of mancozeb was also recorded, by increase in the levels of reactive oxygen species (ROS) generation. Our results demonstrated that ROS plays a critical role in the initiation of mancozeb induced apoptosis in CHLs through two ways, primarily through mitochondria-mediated pathway including induction of ROS, decrease in mitochondrial membrane potential (ΔΨm), along with cytochrome c release from mitochondria, and activation of the caspase cascade. The other pathway includes increase in ROS, which resulted in activation of NF-κB, expression of FasL and triggered FasL-dependent pathway, which also involves caspase-8. Therefore, exposure to mancozeb can lead to induction of apoptosis in CHLs through both mechanisms. Significance: The results of study confirm that mancozeb exposure can induce genotoxicity and apoptosis in CHLs, thus pose a potential risk to exposed human population. © 2012 Elsevier Inc. All rights reserved.
Retraction notice to “Allethrin-induced genotoxicity and oxidative stress in Swiss albino mice” [MUTGEN 747 (1) (2012) 22–28](S1383571812000733)(10.1016/j.mrgentox.2012.03.003)
(Elsevier B.V., 2020) Amit Kumar Srivastava; Pramod Kumar Srivastava; Abdulaziz A. Al-Khedhairy; Javed Musarrat; Yogeshwer Shukla
This article has been retracted: please see Elsevier Policy on Article Withdrawal (https://www.elsevier.com/about/our-business/policies/article-withdrawal). This article has been retracted at the request of the Editor-in-Chief. In this article Figure 2 is described as depicting flow cytometry data of micronuclei induced by allethrin at different doses, but the figure contains duplications which affect the conclusions of the article. The authors have not been able to provide any justification for this image manipulation. As such this article represents a misuse of the scientific publishing system. The scientific community takes a very strong view on this matter and apologies are offered to readers of the journal that this was not detected during the submission process. © 2019
Retraction notice to “Mancozeb-induced genotoxicity and apoptosis in cultured human lymphocytes” [Life Sci. 90/21-22 (2012) 815 - 824] (Life Sciences (2012) 90(21–22) (815–824), (S0024320512000021), (10.1016/j.lfs.2011.12.013))
(Elsevier Inc., 2022) Amit Kumar Srivastava; Wahid Ali; Richa Singh; Kulpreet Bhui; Shilpa Tyagi; Abdulaziz A. Al-Khedhairy; Pramod Kumar Srivastava; Javed Musarrat; Yogeshwer Shukla
This article has been retracted: please see Elsevier Policy on Article Withdrawal (https://www.elsevier.com/about/our-business/policies/article-withdrawal). This article has been retracted at the request of the Editor-in-Chief. Multiple figures in this article appear to be falsified/fabricated. Fig. 2 shows manipulated dot plots for two different treatment groups (2 μl/ml and 5 μl/ml). Fig. 6 (caspase 9 blot) is very similar to fig. 5 (total ERK1/2) of a previous paper from this group published in Cancer Letters, 282, (2009) 67–176, https://doi.org/10.1016/j.canlet.2009.03.003. In Fig. 4, Lanes 4 and 5 are identical for most of their lengths, with only cosmetic changes along the edges of the upper half. Fig. 6 contains similar portions of Fig. 2(a) of a paper published by the same group in Food and Chemical Toxicology, 49, (2011) 1511–1520, https://doi.org/10.1016/j.fct.2011.03.040 The authors have not been able to provide any justification for this image manipulation. © 2022 Elsevier Inc.
Rodent models and contemporary molecular techniques: Notable feats yet incomplete explanations of Parkinson's disease pathogenesis
(Humana Press Inc., 2012) Sharawan Yadav; Anubhuti Dixit; Sonal Agrawal; Ashish Singh; Garima Srivastava; Anand Kumar Singh; Pramod Kumar Srivastava; Om Prakash; Mahendra Pratap Singh
Rodent models and molecular tools, mainly omics and RNA interference, have been rigorously used to decode the intangible etiology and pathogenesis of Parkinson's disease (PD). Although convention of contemporary molecular techniques and multiple rodent models paved imperative leads in deciphering the role of putative causative factors and sequential events leading to PD, complete and clear-cut mechanisms of pathogenesis are still hard to pin down. The current article reviews the implications and pros and cons of rodent models and molecular tools in understanding the molecular and cellular bases of PD pathogenesis based on the existing literature. Probable rationales for short of comprehensive leads and future possibilities in spite of the extensive applications of molecular tools and rodent models have also been discussed. © Springer Science+Business Media, LLC 2012.
Role of metabolites and significance of SH groups in the action of NADP+-linked isocitrate dehydrogenase of urdbean seeds (Phaseolus mungo L.)
(2011) Pramod Kumar Srivastava; Govind Kant Srivastava; Indra Mani; Sharawan Yadav; Asha Anand
NADP+-linked-isocitrate dehydrogenase (EC 1.1.1.42) is a key enzyme of the Tricarboxylic Acid Cycle (TCA) and has been purified from urdbean seeds and it is inhibited by ATP in a competitive manner having inhibitor constant (Ki 1.32 mM. Phosphoenol-pyruvate, an energy rich compound plays an important role in the regulation of this enzyme and this metabolite inhibited the enzyme activity of NADP+-linked-isocitrate dehydrogenase of urdbean with inhibitor constant (Ki 2.66 mM in a competitive manner. The mode of inhibition by various metabolites of Krebs cycle has been carried out and found that oxaloacetate and succinate inhibit ICDH urdbean enzyme in a competitive manner with respect to isocitrate and their Ki values are found to be 7.27 and 10.67 mM, respectively. Citrate inhibits the urdbean ICDH enzyme non competitively with Ki revalue equal to 3.33 mM. The SH groups play a important role in the activity of NADP+-linked-isocitrate dehydrogenase and blocking of this group with SH-reagents, leads to inactivation of urdbean ICDH enzyme. With excess iodoacetamide (1.00 mM) and N-ethylmaleimide (4.0 mM) inhibition of this enzyme follows first order kinetics, suggesting that there are four reactive SH groups per mole of enzyme which are equally reactive and there is no site- site interaction among the tetrameric isoicitrate dehydrogenase of urdbean. © 2011 Academic Journals Inc.
Role of secondary mediators in caffeine-mediated neuroprotection in maneb- and paraquat-induced Parkinson's disease phenotype in the mouse
(2012) Sharawan Yadav; Satya Prakash Gupta; Garima Srivastava; Pramod Kumar Srivastava; Mahendra Pratap Singh
Maneb and paraquat are known to induce Parkinson's disease (PD) phenotype, however, caffeine offers neuroprotection. Nitric oxide (NO) acts an important mediator in PD phenotype and tyrosine kinase (TK), nuclear factor kappa B (NF-kB), p38 mitogen activated protein kinase (p38 MAPK) are known to regulate its production. The present study aimed to elucidate the role of caffeine in the regulation of NO production and microglial activation and their subsequent contribution in dopaminergic neuroprotection. The animals were treated with caffeine and/or maneb and paraquat along with controls. In a few sets of experiments, the animals were also treated with aminoguanidine, an inhibitor of inducible NO synthase, pyrrolidine dithiocarbamate (PDTC), an inhibitor of NF-kB, genistein, an inhibitor of TK or SB202190, an inhibitor of p38 MAPK. Tyrosine hydroxylase (TH)-immunoreactivity and anti-integrin αM (OX-42) staining were performed to assess the number of dopaminergic neurons and activation of microglia, respectively. NO was measured in terms of nitrite, however, the expressions of p38 MAPK, interleukin (IL)-1β, NF-kB and TK were checked by western blot analyses. Maneb and paraquat induced the number of degenerating dopaminergic neurons, microglial cells, nitrite content, expressions of IL-1β, p38 MAPK, NF-kB and TK and caffeine co-treatment reduced the level of such alterations. Reductions were more pronounced in the animals co-treated with aminoguanidine, PDTC, genistein or SB202190. The results obtained thus demonstrate that caffeine down-regulates NO production, neuroinflammation and microglial activation, which possibly contribute to neuroprotection. © Springer Science+Business Media, LLC 2011.
The inhibitory effect of metals and other ions on acid phosphatase activity from vigna aconitifolia seeds
(Taylor and Francis Inc., 2015) Pramod Kumar Srivastava; Asha Anand
Sensitivity of acid phosphatase from Vigna aconitifolia seeds to metal ions, fluoride, and phosphate was examined. All the effectors had different degree of inhibitory effect on the enzyme. Among metal ions, molybdate and ferric ion were observed to be most potent inhibitors and both exhibited mixed type of inhibition. Acid phosphatase activity was inhibited by Cu2+ in a noncompetitive manner. Zn and Mn showed mild inhibition on the enzyme activity. Inhibition kinetics analysis explored molybdate as a potent inhibitor for acid phosphatase in comparison with other effectors used in this study. Fluoride was the next most strong inhibitor for the enzyme activity, and caused a mixed type of inhibition. Phosphate inhibited the enzyme competitively, which demonstrates that inhibition due to phosphate is one of the regulatory factors for enzyme activity. © 2015 Taylor and Francis Group, LLC.
Understanding the language of vitamin C
(2009) Nandini Sarkar; Pramod Kumar Srivastava; Vikash Kumar Dubey
Vitamin C (L-ascorbate) is a good antioxidant. Because of its water soluble nature it can work both inside and outside the cells to combat free radical damage. It has several applications starting from application in cancer therapeutics to treatment of common cold. Human, primates and guinea pigs can not synthesize this nutrient and must have to take this nutrient with diet. The current review brings together information available about the applications of Vitamin C with emphasis on antioxidant property and application in cancer therapy. The contradicting reports about application of Vitamin C in cancer therapy are also discussed. © 2009 Bentham Science Publishers Ltd.