Search results for: protein unfolding
869 Autophagy Regulates Human Hepatocellular Carcinoma Tumorigenesis through Selective Degradation of Cyclin D1
Authors: Shan-Ying Wu, Sheng-Hui Lan, Xi-Zhang Lin, Ih-Jen Su, Ting-Fen Tsai, Chia-Jui Yen, Tsung-Hsueh Lu, Fu-Wen Liang, Huey-Jen Su, Chun-Li Su, Hsiao-Sheng Liu
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In hepatocelluar carcinoma (HCC), dysregulated expression of cyclin D1 and impaired autophagy has been reported separately. However, the relationship between them has not been explored. In this study, we demonstrated that autophagy was inversely correlated with cyclin D1 expression in 147 paired HCC patient specimens. HCC specimen with highly expression of cyclin D1 shows correlation with poor overall survival rate. Furthermore, induction of autophagy by amiodarone (antiarrhythmic drug) in Hep 3B cells, cyclin D1 was recruited into autophagosomes demonstrated by immune-gold labeling of cyclin D1 after extraction of autophagosomes. We further demonstrated that autophagy suppresses Hep 3B cell proliferation, and further analysis revealed that cell cycle was arrested at G1 phase. The interaction between LC3 (maker of autophagy) and cyclin D1 was increased after autophagy induction. In addition, ubiquitinated-cyclin D1 was also increased after autophagy induction, which is selectively degraded by autophagosome through binding with SQSTM1/p62 (an adaptor protein). In vivo study showed that amiodarone induced autophagy suppresses liver tumor formation in xenograft mouse and orthotopic rat model through decreasing cyclin D1 expression and inhibition of cell proliferation. Altogether, we reveal a novel mechanism that ubiquitinated cyclin D1 degraded by autophagic pathway by p62 and amiodarone is a promising drug for targeting cyclin D1 in liver cancer therapy.Keywords: autophagy, cyclin D1, hepatocellular carcinoma, amiodarone
Procedia PDF Downloads 295868 Phylogenetic Analyses of Newcastle Disease Virus Isolated from Unvaccinated Chicken Flocks in Kyrgyzstan from 2015 to 2016
Authors: Giang Tran Thi Huong, Hieu Dong Van, Tung Dao Duy, Saadanov Iskender, Isakeev Mairambek, Tsutomu Omatsu, Yukie Katayama, Tetsuya Mizutani, Yuki Ozeki, Yohei Takeda, Haruko Ogawa, Kunitoshi Imai
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Newcastle disease virus (NDV) is a contagious viral disease of the poultry industry and other birds throughout the world. At present, very little is known about molecular epidemiological data regarding the causes of ND outbreak in commercial poultry farms in Kyrgyzstan. In the current study, the NDV isolated from the one out of three samples from the unvaccinated flock was confirmed as NDV. Phylogenetic analysis indicated that this NDV strain is clustered in the Class II subgenotype VIId, and closely related to the Chinese NDV isolate. Phylogenetic analyses revealed that the isolated NDV strain has an origin different from the 4 NDV strains previously identified in Kyrgyzstan. According to the mean death time (MDT: 61.1 h) and a multibasic amino acid (aa) sequence at the F0 proteolytic cleavage site (¹¹²R-R-Q-K-R-F¹¹⁷), the NDV isolate was determined as mesogenic strain. Several mutations in the neutralizing epitopes (notably, ³⁴⁷E→K) and the global head were observed in the hemagglutinin-neuraminidase (HN) protein of the current isolate. The present study represents the molecular characterization of the coding gene region of NDV in Kyrgyzstan. Additionally, further study will be investigated on the antigenic characterization using monoclonal antibody.Keywords: Kyrgyzstan, Newcastle disease, genotype, genome characterization
Procedia PDF Downloads 142867 The Interplay of Dietary Fibers and Intestinal Microbiota Affects Type 2 Diabetes by Generating Short-Chain Fatty Acids
Authors: Muhammad Mazhar, Yong Zhu, Likang Qin
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Foods contain endogenous components known as dietary fibers, which are classified into soluble and insoluble forms. Dietary fibers are resistant to gut digestive enzymes, modulating anaerobic intestinal microbiota (AIM) and fabricating short-chain fatty acids (SCFAs). Acetate, butyrate, and propionate dominate in the gut, and different pathways, including Wood-Ljungdahl and acrylate pathways, generate these SCFAs. In pancreatic dysfunction, the release of insulin/glucagon is impaired, which leads to hyperglycemia. SCFAs enhance insulin sensitivity or secretion, beta-cell functions, leptin release, mitochondrial functions, and intestinal gluconeogenesis in human organs, which positively affect type 2 diabetes (T2D). Research models presented that SCFAs either enhance the release of peptide YY (PYY) and glucagon-like peptide-1 (GLP-1) from L-cells (entero-endocrine) or promote the release of leptin hormone satiation in adipose tissues through G-protein receptors, i.e., GPR-41/GPR-43. Dietary fibers are the components of foods that influence AIM and produce SCFAs, which may be offering beneficial effects on T2D. This review addresses the effectiveness of SCFAs in modulating gut AIM in the fermentation of dietary fiber and their worth against T2D.Keywords: dietary fibers, intestinal microbiota, short-chain fatty acids, fermentation, type 2 diabetes
Procedia PDF Downloads 73866 Effect of Erythropoietin Hormone Supplementation on Hypoxia-Inducible Factor1-Alpha in Rat Kidneys with Experimental Diabetic Nephropathy
Authors: Maha Deif, Alaa Eldin Hassan, Eman Shaat, Nesrine Elazhary, Eman Magdy
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Background: Erythropoietin (EPO) is a hematopoietic factor with multiple protective effects. The aim of the present study was to investigate the potential effect of EPO administration on renal functions and hypoxia inducible factor 1-alpha (HIF-1a) in diabetic rat kidneys. Methodology: The current study was carried out on 40 male albino rats divided into four groups (n= 10 in each). Group I served as normal control, group II was the diabetic control, group III rats received EPO on the same day of diagnosis of diabetes mellitus (DM), while group IV received the first dose of EPO 2 weeks after the diagnosis of DM. Results: The results showed that EPO supplementation leads to a significant decrease in serum urea, urinary protein and creatinine clearance as well as a significant increase in renal HIF-1a in group III and IV rats compared to the diabetic control group (group II). However, fasting blood glucose was significantly decreased in group III as compared to the diabetic control group in the third week, but no significant difference was reported in the fourth week among groups II, III and IV. Conclusion: EPO administration leads to the improvement of renal functions and increased levels of HIF-1a in diabetic rats.Keywords: erythropoietin, diabetic nephropathy, hypoxia-inducible factor1-alpha, renal functions
Procedia PDF Downloads 286865 Simultaneous Targeting of MYD88 and Nur77 as an Effective Approach for the Treatment of Inflammatory Diseases
Authors: Uzma Saqib, Mirza S. Baig
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Myeloid differentiation primary response protein 88 (MYD88) has long been considered a central player in the inflammatory pathway. Recent studies clearly suggest that it is an important therapeutic target in inflammation. On the other hand, a recent study on the interaction between the orphan nuclear receptor (Nur77) and p38α, leading to increased lipopolysaccharide-induced hyperinflammatory response, suggests this binary complex as a therapeutic target. In this study, we have designed inhibitors that can inhibit both MYD88 and Nur77 at the same time. Since both MYD88 and Nur77 are an integral part of the pathways involving lipopolysaccharide-induced activation of NF-κB-mediated inflammation, we tried to target both proteins with the same library in order to retrieve compounds having dual inhibitory properties. To perform this, we developed a homodimeric model of MYD88 and, along with the crystal structure of Nur77, screened a virtual library of compounds from the traditional Chinese medicine database containing ~61,000 compounds. We analyzed the resulting hits for their efficacy for dual binding and probed them for developing a common pharmacophore model that could be used as a prototype to screen compound libraries as well as to guide combinatorial library design to search for ideal dual-target inhibitors. Thus, our study explores the identification of novel leads having dual inhibiting effects due to binding to both MYD88 and Nur77 targets.Keywords: drug design, Nur77, MYD88, inflammation
Procedia PDF Downloads 305864 Developing Drought and Heat Stress Tolerant Chickpea Genotypes
Authors: Derya Yucel, Nigar Angın, Dürdane Mart, Meltem Turkeri, Volkan Catalkaya, Celal Yucel
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Chickpea (Cicer arietinum L.) with high protein content is a vital food, especially in under-developed and developing countries for the people who do not consume enough meat due to low-income level. The objective of the proposed study is to evaluate growing, yield and yield components of chickpea genotypes under Mediterranean condition so determine tolerance of chickpea genotypes against drought and heat stress. For this purpose, a total of 34 chickpea genotypes were used as material. The experiment was conducted according to factorial randomized complete block design with 3 reps at the Eastern Mediterranean Research Institute, Adana, TURKEY for 2014-15 growing season under three different growing conditions (Winter sowing, irrigated-late sowing and non-irrigated- late sowing). According to results of this experiment, vegetative period, flowering time, poding time, maturity time, plant height, height of first pod, seed yield and 100 seed weight were ranged between 68.33 to 78.77 days, 94.22 to 85.00 days, 94.11 to 106.44 days, 198.56 to 214.44 days, 37.18 to 64.89 cm, 18.33 to 34.83 cm, 417.1 to 1746.4 kg/ha and 14.02 to 45.02 g, respectively. Among the chickpea genotypes, the Aksu, Arda, Çakır, F4 09 (X 05 TH 21-16189), FLIP 03-108 were least affected by drought and heat stress. Therefore, these genotypes can be used as sources of drought and heat tolerance in further breeding programme for evolving the drought and heat tolerant genotypes in chickpea.Keywords: chickpea, drought stress, heat stress, yield
Procedia PDF Downloads 229863 Effect of Nicorandil in Bile Duct Ligation-Induced Liver Fibrosis in Rats: Role of Hepatic Stellate Cells
Authors: Y. S. Mohamed, L. A. Ahmed, H. A. Salem, A. M. Agha
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Liver Fibrosis is one of the most serious conditions that affect the Egyptian society. In the present study, the effect of nicorandil was investigated in experimentally-induced liver fibrosis by bile duct ligation in rats. Nicorandil (3mg/kg/day) was given orally 24 h after bile duct ligation for 14 days till the end of the experiment. Nicorandil group showed a significant improvement in liver function tests (ALT and ALP) as well as a significant decrease in oxidative stress biomarkers (TBARS and GSH), area of fibrosis and activity of hepatic stellate cells as indicated by decreased expression of alpha smooth muscle actin.Moreover, nicorandil treatment decreased HSCs proliferation due to its inhibitory effects on protein kinase C(PKC) and Platelet derived growth factor (PDGF) . Oral administration of either glibenclamide (10 mg/kg/day)(a KATP channel blocker) or L-NAME (30 mg/kg/day) (an inhibitor of nitric oxide synthase) blocked the protective effects of nicorandil. However, nicorandil and L-NAME treated group showed more or less results similar to that of untreated bile duct ligated group. In conclusion, nicorandil was effective against the development of bile duct ligated-induced liver fibrosis in rats where activation of the NO pathway plays an important role in the protective effect nicorandil.Keywords: hepatic stellate cells, nicorandil, nitric oxide donor, liver fibrosis
Procedia PDF Downloads 611862 Sustainable Improvement in Soil Properties and Maize Performance by Organic Fertilizers at Different Levels
Authors: Shahid Iqbal, Haroon Z. Khan, Muhammad Arif
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A sustainable agricultural system involving the improvement in soil properties and crop performance cannot be developed without organic fertilizer use. The effects of poultry manure compost (PMC) and pressmud compost (PrMC) at different levels on improving the soil properties and maize performance has not been yet described by any study comprehensively. Thus, field experiments (2011 and 2012) were conducted at Agronomy Research Area, University of Agriculture Faisalabad (31°26'5" N and 73°4'6" E) in sandy loam soil to determine the improvement in soil properties and maize performance due to application of PMC and PrMC each at five different levels (2, 4, 6, 8 and 10 t ha-1). A control (unamended) treatment was also included for comparison. The results indicated that performance of PMC levels was superior to PrMC levels. Increasing both composts levels improved soil properties, maize growth, and stover yield. Results showed that during both years’ highest rates of PMC i.e. 10 and 8 t ha-1 improved the soil properties: ECe, pH, inorganic N, OM, and WHC higher than other treatments. While, 10 and 8 t PMC ha-1 also significantly increased leaf area index (LAI), crop growth rate (CGR) and net assimilation rate (NAR), and stover yield. Similarly, 10 and 8 t PMC ha-1 also improved the grain protein content, but contrarily, grain oil was lowest for 10 and 8 t ha-1 PMC during both years. Moreover, in both years highest gross and net income, and benefit cost ratio was also achieved by 10 and 8 t ha-1 PMC. It is concluded that PMC at rate of 10 and 8 t ha-1 sustainably improved soil properties and maize performance.Keywords: compost, soil, maize, growth, yield
Procedia PDF Downloads 364861 Feasibility of Chicken Feather Waste as a Renewable Resource for Textile Dyeing Processes
Authors: Belayihun Missaw
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Cotton cationization is an emerging area that solves the environmental problems associated with the reactive dyeing of cotton. In this study, keratin hydrolysate cationizing agent from chicken feather was extracted and optimized to eliminate the usage of salt during dyeing. Cationization of cotton using the extracted keratin hydrolysate and dyeing of the cationized cotton without salt was made. The effect of extraction parametric conditions like concentration of caustic soda, temperature and time were studied on the yield of protein from chicken feather and colour strength (K/S) values, and these process conditions were optimized. The optimum extraction conditions were. 25g/l caustic soda, at 500C temperature and 105 minutes with average yield = 91.2% and 4.32 colour strength value. The effect of salt addition, pH and concentration of cationizing agent on yield colour strength was also studied and optimized. It was observed that slightly acidic condition with 4% (% owf) concentration of cationizing agent gives a better dyeability as compared to normal cotton reactive dyeing. The physical properties of cationized-dyed fabric were assessed, and the result reveals that the cationization has a similar effect as normal dyeing of cotton. The cationization of cotton with keratin extract was found to be successful and economically viable.Keywords: cotton materials, cationization, reactive dye, keratin hydrolysate
Procedia PDF Downloads 63860 Chitosan Modified Halloysite Nanomaterials for Efficient and Effective Vaccine Delivery in Farmed Fish
Authors: Saji George, Eng Khuan Seng, Christof Luda
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Nanotechnology has been recognized as an important tool for modern agriculture and has the potential to overcome some of the pressing challenges faced by aquaculture industry. A strategy for optimizing nanotechnology-based therapeutic delivery platform for immunizing farmed fish was developed. Accordingly, a compositional library of nanomaterials of natural chemistry (Halloysite (clay), Chitosan, Hydroxyapatite, Mesoporous Silica and a composite material of clay-chitosan) was screened for their toxicity and efficiency in delivering models antigens in cellular and zebrafish embryo models using high throughput screening platforms. Through multi-parametric optimization, chitosan modified halloysite (clay) nanomaterial was identified as an optimal vaccine delivery platform. Further, studies conducted in juvenile seabass showed the potential of clay-chitosan in delivering outer membrane protein of Tenacibaculum maritimum- TIMA (pathogenic bacteria) to and its efficiency in eliciting immune responses in fish. In short, as exemplified by this work, the strategy of using compositional nanomaterial libraries and their biological profiling using high-throughput screening platform could fasten the discovery process of nanomaterials with potential applications in food and agriculture.Keywords: nanotechnology, fish-vaccine, drug-delivery, halloysite-chitosan
Procedia PDF Downloads 282859 Phosphoinositide 3-Kinase-Dependent CREB Activation is Required for the Induction of Aromatase in Tamoxifen-Resistant Breast Cancer
Authors: Ji Hye Im, Nguyen T. T. Phuong, Keon Wook Kang
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Estrogens are important for the development and growth of estrogen receptor (ER)-positive breast cancer, for which anti-estrogen therapy is one of the most effective treatments. However, its efficacy can be limited by either de novo or acquired resistance. Aromatase is a key enzyme for the biosynthesis of estrogens, and inhibition of this enzyme leads to profound hypoestrogenism. Here, we found that the basal expression and activity of aromatase were significantly increased in tamoxifen (TAM)-resistant human breast cancer (TAMR-MCF-7) cells compared to control MCF-7 cells. We further revealed that aromatase immunoreactivity in tumor tissues was increased in recurrence group after TAM therapy compared to non-recurrence group after TAM therapy. Phosphorylation of Akt, extracellular signal-regulated kinase (ERK), and p38 kinase were all increased in TAMR-MCF-7 cells. Inhibition of phosphoinositide 3-kinase (PI3K) suppressed the transactivation of the aromatase gene and its enzyme activity. Furthermore, we have also shown that PI3K/Akt-dependent cAMP-response element binding protein (CREB) activation was required for the enhanced expression of aromatase in TAMR-MCF-7 cells. Our findings suggest that aromatase expression is up-regulated in TAM-resistant breast cancer via PI3K/Akt-dependent CREB activation.Keywords: TAMR-MCF-7, CREB, estrogen receptor, aromatase
Procedia PDF Downloads 412858 Genome Sequencing of the Yeast Saccharomyces cerevisiae Strain 202-3
Authors: Yina A. Cifuentes Triana, Andrés M. Pinzón Velásco, Marío E. Velásquez Lozano
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In this work the sequencing and genome characterization of a natural isolate of Saccharomyces cerevisiae yeast (strain 202-3), identified with potential for the production of second generation ethanol from sugarcane bagasse hydrolysates is presented. This strain was selected because its capability to consume xylose during the fermentation of sugarcane bagasse hydrolysates, taking into account that many strains of S. cerevisiae are incapable of processing this sugar. This advantage and other prominent positive aspects during fermentation profiles evaluated in bagasse hydrolysates made the strain 202-3 a candidate strain to improve the production of second-generation ethanol, which was proposed as a first step to study the strain at the genomic level. The molecular characterization was carried out by genome sequencing with the Illumina HiSeq 2000 platform paired end; the assembly was performed with different programs, finally choosing the assembler ABYSS with kmer 89. Gene prediction was developed with the approach of hidden Markov models with Augustus. The genes identified were scored based on similarity with public databases of nucleotide and protein. Records were organized from ontological functions at different hierarchical levels, which identified central metabolic functions and roles of the S. cerevisiae strain 202-3, highlighting the presence of four possible new proteins, two of them probably associated with the positive consumption of xylose.Keywords: cellulosic ethanol, Saccharomyces cerevisiae, genome sequencing, xylose consumption
Procedia PDF Downloads 320857 Bioinformatic Study of Follicle Stimulating Hormone Receptor (FSHR) Gene in Different Buffalo Breeds
Authors: Hamid Mustafa, Adeela Ajmal, Kim EuiSoo, Noor-ul-Ain
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World wild, buffalo production is considered as most important component of food industry. Efficient buffalo production is related with reproductive performance of this species. Lack of knowledge of reproductive efficiency and its related genes in buffalo species is a major constraint for sustainable buffalo production. In this study, we performed some bioinformatics analysis on Follicle Stimulating Hormone Receptor (FSHR) gene and explored the possible relationship of this gene among different buffalo breeds and with other farm animals. We also found the evolution pattern for this gene among these species. We investigate CDS lengths, Stop codon variation, homology search, signal peptide, isoelectic point, tertiary structure, motifs and phylogenetic tree. The results of this study indicate 4 different motif in this gene, which are Activin-recp, GS motif, STYKc Protein kinase and transmembrane. The results also indicate that this gene has very close relationship with cattle, bison, sheep and goat. Multiple alignment (MA) showed high conservation of motif which indicates constancy of this gene during evolution. The results of this study can be used and applied for better understanding of this gene for better characterization of Follicle Stimulating Hormone Receptor (FSHR) gene structure in different farm animals, which would be helpful for efficient breeding plans for animal’s production.Keywords: buffalo, FSHR gene, bioinformatics, production
Procedia PDF Downloads 532856 Investigating the Role of Dystrophin in Neuronal Homeostasis
Authors: Samantha Shallop, Hakinya Karra, Tytus Bernas, Gladys Shaw, Gretchen Neigh, Jeffrey Dupree, Mathula Thangarajh
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Abnormal neuronal homeostasis is considered a structural correlate of cognitive deficits in Duchenne Muscular Dystrophy. Neurons are highly polarized cells with multiple dendrites but a single axon. Trafficking of cellular organelles are highly regulated, with the cargo in the somatodendritic region of the neuron not permitted to enter the axonal compartment. We investigated the molecular mechanisms that regular organelle trafficking in neurons using a multimodal approach, including high-resolution structural illumination, proteomics, immunohistochemistry, and computational modeling. We investigated the expression of ankyrin-G, the master regulator controlling neuronal polarity. The expression of ankyrin G and the morphology of the axon initial segment was profoundly abnormal in the CA1 hippocampal neurons in the mdx52 animal model of DMD. Ankyrin-G colocalized with kinesin KIF5a, the anterograde protein transporter, with higher levels in older mdx52 mice than younger mdx52 mice. These results suggest that the functional trafficking from the somatodendritic compartment is abnormal. Our data suggests that dystrophin deficiency compromised neuronal homeostasis via ankyrin-G-based mechanisms.Keywords: neurons, axonal transport, duchenne muscular dystrophy, organelle transport
Procedia PDF Downloads 95855 Dietary Pattern and Risk of Breast Cancer Among Women:a Case Control Study
Authors: Huma Naqeeb
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Epidemiological studies have shown the robust link between breast cancer and dietary pattern. There has been no previous study conducted in Pakistan, which specifically focuses on dietary patterns among breast cancer women. This study aims to examine the association of breast cancer with dietary patterns among Pakistani women. This case-control research was carried in multiple tertiary care facilities. Newly diagnosed primary breast cancer patients were recruited as cases (n = 408); age matched controls (n = 408) were randomly selected from the general population. Data on required parameters were systematically collected using subjective and objective tools. Factor and Principal Component Analysis (PCA) techniques were used to extract women’s dietary patterns. Four dietary patterns were identified based on eigenvalue >1; (i) veg-ovo-fish, (ii) meat-fat-sweet, (iii) mix (milk and its products, and gourds vegetables) and (iv) lentils - spices. Results of the multiple regressions were displayed as adjusted odds ratio (Adj. OR) and their respective confidence intervals (95% CI). After adjusted for potential confounders, veg-ovo-fish dietary pattern was found to be robustly associated with a lower risk of breast cancer among women (Adj. OR: 0.68, 95%CI: (0.46-0.99, p<0.01). The study findings concluded that attachment to the diets majorly composed of fresh vegetables, and high quality protein sources may contribute in lowering the risk of breast cancer among women.Keywords: breast cancer, dietary pattern, women, principal component analysis
Procedia PDF Downloads 123854 Use of High Hydrostatic Pressure as an Alternative Preservation Method for Fresh Dates, Rutab
Authors: Salah Mohammed Al-Eid, Siddig Hussein Hamad, Fahad Mohammed Aljassas
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The effects of high hydrostatic pressure (HHP) treatments on microbial contamination, chemical and physical properties of fresh dates (Rutab stage) were studied. Khalas, Barhi and Hilali cultivars were treated at 200, 250, 300 and 350 MPa using HHP research apparatus. The objective of such treatments was to preserve fresh dates without adversely affecting its properties. Treating fresh dates at 300 MPa for 5 minutes at 40°C reduced microbial contamination in about 2.5 log cycles. Applying 250 MPa was enough to control Rutab contamination with molds, yeasts, and coliforms. Both treatments were enough to reduce Rutab microbial contamination to acceptable levels. HHP caused no significant effect on Rutab chemical properties (moisture, sugars, protein, pectin and acidity). However, a slight decrease in moisture contents due to HHP was observed. Rutab lightness (L*) significantly decreased due to the application of HHP. Only Rutab treated at 300 MPs gave lower redness (a*) values compared with an untreated sample. The effect of 300 MPa on increasing yellowness (b*) was observed for Barhi and Hilali but decreasing for Khalas. The hardness of all Rutab cultivars significantly decreased as a result of HHP application. In fact, the pressure applied at 300 MPa had an adverse effect on texture, which may limit its suitability for use in Rutab preservation.Keywords: high hydrostatic pressure, fresh dates (Rutab), microbial contamination, color, texture
Procedia PDF Downloads 293853 The Transcriptional Regulation of Human LRWD1 through DNA Methylation
Authors: Yen-Ni Teng, Hsing-Yi Chen, Hsien-An Pan, Yung-Ming Lin, Hany A. Omar, Jui-Hsiang Hung
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Leucine-rich repeats and WD repeat domain containing 1 (LRWD1) is highly expressed in the testes of healthy males. On the other hand, LRWD1 is significantly down-regulated in the testicular tissues of patients with severe spermatogenic defects. In our study, the downregulation of LRWD1 expression by shRNA caused a significant reduction of cell growth and mitosis and a noteworthy increase in the cell microtubule atrophy rate. Here, we used EMBOSS CpG plot analysis to explore the promoter region of LRWD1 gene. We found that CpG islands are located between positions -253 to +5 nucleotides upstream from the LRWD1 transcription start site. Luciferase reporter assay revealed that the hypermethylation of the LRWD1 promoter reduced the transcription activity in cells. In addition, quantitative methylation-specific PCR and immunostaining showed that the methylation inhibitor, 5-Aza-2'-deoxycytidine, increased LRWD1 promoter activity, LRWD1 mRNA, protein expression and cell viability. Whereas, the methylation activator, S-adenosylmethionine, caused opposite effects. The overexpression of p53 and Nrf2 in NT2/D1 cells increased LRWD1 promoter activity while 5-fluorodeoxyuridine decreased it. In conclusion, this study highlights evidence that the methylation status of LRWD1 promoter is associated with LRWD1 expression. Since the expression level of LRWD1 plays an important role in spermatogenesis, the methylation status of LRWD1 may serve as a novel molecular diagnostic or therapeutic approach in male's infertility.Keywords: LRWD1, DNA methylation, p53, Nrf2
Procedia PDF Downloads 148852 Discovery of New Inhibitors for Colorectal Cancer Treatment
Authors: Kai-Cheng Hsu, Tzu-Ying Sung, Jinn-Moon Yang
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Colorectal cancer (CRC) is one of the main causes of cancer death in the world. Although several drugs have been developed to treat colorectal cancer, such as Regorafenib and 5-FU, their efficacy is often limited by the development of drug resistance. Therefore, development of new drugs with new scaffolds is necessary to treat CRC. Here, we used site-moiety maps to identify inhibitors against PIM1, LIMK1, SRC, and mTOR, which are often overexpressed in CRC. A site-moiety map represents physicochemical properties and moiety preferences of a binding site through anchors. An anchor contains three elements: (1) conserved interacting residues of a binding pocket; (2) moiety preference of the binding pocket; and (3) the type (e.g., hydrogen-bonding or van der Waals interactions) of interaction between the moieties and the binding pocket. Then, we performed a structure-based virtual screening of ~260,000 compounds and selected compound candidates with high site-moiety map scores for bioassays. Among these candidates, compound 1 and compound 2 inhibited the growth of CRC cells with IC50 values of <10 μM. The experimental result of enzyme-based assays indicated that compound 1 is a dual inhibitor against PIM1 (IC50 6 μM) and LIMK1(IC50 11 μM). Compound 2 was predicted as a SRC inhibitor and will be further validated. The compounds inhibited different protein targets compared to the current drugs. We believe that the compounds provide a starting point to design new drugs for CRC treatment.Keywords: colorectal cancer, drug discovery, site-moiety map, virtual screening, PIM1, LIMK1
Procedia PDF Downloads 246851 Effect of Sweet Potato (Ipomoea batatas) Leaves on Wheat Offal Replacement for Chicks Feed Production
Authors: C. C. Okafor, T. M. Ezeh
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The effect of addition of sweet potato leaves in replacement of wheat offal in the production of broiler chicks feed was studied. 72 day-old marshal strain chicks were used and brooded for two weeks with a normal commercial feed in Nigeria called top feed and weighed separately at the end of the two weeks, complete randomized design (CRD) was used. The weighed broiler chicks were randomly allocated to four dietary treatments. Each treatment was replicated to twice with eighteen birds per replicate. The four dietary treatment identified as T1, T2, T3 and T4. T1 served as control diet with 21% crude protein content, while T2 was prepared with Enzyme and in T3 and T4, wheat offal was replaced with sweet potato leaves and in T4 with inclusion of enzyme. Growth performance was studied using the following daily feed intake, daily weight gain and feed efficiency. The result in daily weight gain showed that chicks fed with T2 feed had the highest weight gain (93.75) while chicks fed with T3 had the least weight gain of (34.5 gm). In daily feed intake chicks fed with T4 fed more (53.06 gm) than chicks fed with T2 (51.08 gm). In feed efficiency T3 had the highest value of 30% while the T2 had the least efficiency of 22%. There was no significant difference (P≥ 0.05) in all the three parameter tested. Sweet potato leaves can replace wheat offal in broiler feed production without any adverse effect on the growth performance.Keywords: broiler, diet, dietary, potato leaves, wheat offal
Procedia PDF Downloads 524850 Nutritional Composition of Crackers Produced from Blend of Sprouted Pigeon Pea (Cajanus cajan), Unripe Plantain (Musa parasidiaca), and Brewers’ Spent Grain Flour and Blood Glucose Level of Diabetic Rats Fed the Biscuit
Authors: Nneka N. Uchegbu, Charles N. Ishiwu
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The nutritional composition and hypoglycaemic effect of crackers produced from a blend of sprouted pigeon pea, unripe plantain, and brewers’ spent grain and fed to Alloxan induced diabetic rat was investigated. Crackers were produced from different blends of sprouted pigeon pea, unripe plantain and brewers’ spent grain. The crackers were evaluated for proximate composition, amino acid profile and antinutritional factors. Blood glucose levels of normal and diabetic rats fed with the control sample and different formulations of cracker were measured. The protein content of the samples were significantly different (p < 0.05) from each other with sample A having the lowest value and sample B with the highest value. The values obtained showed that the samples contained most of the amino acids that are found in plant proteins. The levels of antinutritional factor determined were generally low. Administration of the formulated cracker meals led to a significant reduction in the fasting blood glucose level in the diabetic rats. The present study concluded that consumption of crackers produced from this composite flour can be recommended for the diabetics and those who are sceptical about the disease.Keywords: crackers, diabetics rat, sprouted pigeon pea, unripe plantain and brewers’ spent grain
Procedia PDF Downloads 440849 Use of Apple Pomace as a Source of Dietary Fibre in Mutton Nuggets
Authors: Aamina B. Hudaa, Rehana Akhtera, Massarat Hassana, Mir Monisab
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Mutton nuggets produced with the addition of apple pomace at the levels of 0% (Control), 5% (Treatment 1), 10% (Treatment 2), and 15% (Treatment 3) were evaluated for emulsion stability, cooking yield, pH, proximate composition, texture analysis and sensory properties. Apple pomace addition resulted in significantly higher (p ≤ 0.05) emulsion stability and cooking yield of treatments in comparison to control and pH values were significantly higher (p ≤ 0.05) for the control as compared to treatments. Among the treatments, the product with 15% apple pomace had significantly (p ≤ 0.05) highest moisture content, and protein, ash and fat contents were significantly (p ≤ 0.05) higher in control than treatment groups. Crude fiber content of control was found significantly (p ≤ 0.05) lower in comparison to nuggets formulated with 5%, 10% and 15% apple pomace and was found to increase significantly (p ≤ 0.05) with the increasing levels of apple pomace. Hardness of the products significantly (p ≤ 0.05) decreased with addition of apple pomace, whereas springiness, cohesiveness, chewiness and gumminess showed a non-significant (p ≥ 0.05) decrease with the levels of apple pomace. Sensory evaluation showed significant (p ≤ 0.05) reduction in texture, flavour and overall acceptability scores of treatment products; however the scores were in the range of acceptability and T-1 showed better acceptability among apple pomace incorporated treatments.Keywords: Mutton nuggets, apple pomace, textural properties, sensory evaluation
Procedia PDF Downloads 324848 Production, Optimization, Characterization, and Kinetics of a Partially Purified Laccase from Pleurotus citrinopileatus and Its Application in Swift Bioremediation of Azo Dyes
Authors: Ankita Kushwaha, M. P. Singh
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Background: In the present investigation the efficiency of laccase (benzenediol: oxygen oxidoreductase, EC 1.10.3.2) from Pleurotus citrinopileatus was assessed for the decolorization of azo dyes. Aim: Enzyme production, characterization and kinetics of a partially purified laccase from Pleurotus citrinopileatus were determined for its application in bioremediation of azo dyes. Methods & Results: Laccase has been partially purified by using 80% ammonium sulphate solution. Total activity, total protein, specific activity and purification fold for partially purified laccase were found to be 40.38U, 293.33mg/100ml, 0.91U/mg and 2.84, respectively. The pH and temperature optima of laccase were 5.0 and 50ºC, respectively, while the enzyme was most stable at pH 4.0 and temperature 30ºC when exposed for one hour. The Km of the partially purified laccase for substrates guaiacol, DMP (2,6-dimethoxyphenol) and syringaldazine (3,5-dimethoxy-4-hydroxybenzaldehyde azine) were 60, 95 and 26, respectively. This laccase has been tested for the use in the bioremediation of azo dyes in the absence of mediator molecules. Two dyes namely congo red and bromophenol blue were tested. Discussion: It was observed that laccase enzyme was very effective in the decolorization of these two dyes. More than 80% decolorization was observed within half an hour even in the absence of mediator and their lower Km value indicates that efficiency of the enzyme is very high. The results were promising due to quicker decolorization in the absence of mediators showing that it can be used as a valuable biocatalyst for quick bioremediation of azo dyes. Conclusion: The enzymatic properties of laccase from P. citrinopileatus should be considered for a potential environmental (biodegradation and bioremediation) or industrial applications.Keywords: azo dyes, decolorization, laccase, P.citrinopileatus
Procedia PDF Downloads 220847 Punica granatum (Pomegranate) of a Libyan Variety Exhibits in vitro Anti-Inflammatory Potential
Authors: Lamees A. Ben Saad, Kah Hwi Kim, Chin Chew Quah, Mustafa Shahimi
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Background: Punica granatum (pomegranate) was used as a traditional medicine in different parts of the world. It has been used in the treatment of pain and inflammatory conditions such as peptic ulcer. The numerous risks associated with nonsteroidal anti-inflammatory drugs (NSAIDs) for the treatment of pain and inflammation give rise to using medicinal herbs as alternative therapies. This study aimed to evaluate the anti-inflammatory effect of the ethyl acetate pomegranate fraction (EtOAc) by determination of its inhibitory effects on lipopolysaccharide (LPS), stimulated nitric oxide (NO), prostaglandin E2 (PGE-2), interleukin-6 (IL-6) and cyclooxxgenase-2 (COX2) release from RAW264.7cells. Methods: The inhibitory effect of EtOAc was evaluated on (LPS) induced NO production, PGE2, and IL-6 quantified by immunoassay kit and prostaglandin E2 competitive ELISA kit. COX2 production is an in vitro indication of possible anti-inflammatory activity and was estimated by Western blotting. Results: EtOAc potentially inhibited LPS-induced nitric oxide, prostaglandin, and IL-6 production. With these findings, it was evident that the EtOAc could reduce the LPS-induced cyclooxygenase-2 (COX-2) at the protein level in a dose-dependent manner as determined by Western blotting. Conclusion: The results emphasize potential therapeutic applications of Punica granatum in the treatment of inflammation.Keywords: inflammation, Punica granatum, cytotoxicity, cytokines
Procedia PDF Downloads 660846 Impact of Saline Water and Water Restriction in Laying Hens
Authors: Reza Vakili
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This experiment was conducted to investigate the effect of duration water restriction of drinking water and salinity level on production performance, egg quality and biochemical and hematological blood indices of laying hens. A total of 240 Hy-Line laying hens were used in a completely randomized design with a 2 × 2 factorial arrangement of treatments. Experimental treatments were: 1) free access to drinking water and a low level of salinity (TDS below 500 mg/L) (FAW+LS), 2) free access to water and a high level of salinity (TDS above 1500 mg/L), (FAW+HS), 3) 12 h nightly water restriction and a low level of salinity (LAW+LS), and 4) 12 h water restriction and a high level of salinity (LAW+HS). Intake of feed, percentage of egg production and egg weight and mass were not affected by water restriction or salinity level (P > 0.05), however, a trend (P < 0.01) for lower water consumption was detected in water-restricted hens, regardless of salinity level (213 vs 187). A tendency for lower eggshell and yolk weights was observed in hens that had limited access to water with high salinity compared to those had free access to high saline water (P = 0.08). Serum total protein and glucose concentrations significantly reduced (P < 0.05) in hens drank high salinity water, regardless of water restriction. Moreover, saline water increased the concentration of uric acid, creatinine, and cholesterol when compared to low salinity drank-hens (P < 0.05). The concentrations of ALT and AST increased with salinity level (P < 0.05) and water restriction caused an increment in AST content (P < 0.05). In conclusion, Hy-Line laying hens could withstand water restriction, whilst could not tolerate water salinity of about 1500 mg/L.Keywords: chemical pollutants, eggs, laying hens, salinity, water quality
Procedia PDF Downloads 23845 The Role of Vitamin D Supplementation in Augmenting IFN-γ Production in Response to Mycobacterium Tuberculosis Infection: A Randomized Controlled Trial
Authors: Muhammad Imran Hussain, Ramisha Ibtisam, Tayyaba Fatima, Huba Khalid, Ayesha Aziz, Khansa, Adan Sitara, Anam Shahzad, Aymen Jabeen
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Vitamin D supports the immune system fight TB by inhibiting Interferon-gamma (IFN-γ) and lowering host inflammation. The purpose of the research was to see if giving the vitamin D supplements to TB patients affected their prognosis. A randomized placebo control study of 200 TB patients was performed among which 106 received 400,000 IU of injectable vitamin D3 and 94 received placebo for 2 doses. Assessment was carried out at the end of every month for 3 months. IFN-γ responses to whole blood stimulation generated by the Mycobacterium tuberculosis sonicate (MTBs) antigen and early secreted and T cell activated 6 kDa (ESAT6) were assessed at 0 and 12 weeks. The statistical analysis used descriptive statistics (mean and standard deviation), Friedman's test and Fisher's test. The vitamin D group gained significantly more weight (+3.90 pounds) and had less persistent lung disease on imaging (1.33 zones vs. 1.84 zones). They also had a 50% decrease in cavity size. Additionally, patients with low baseline serum concentrations of 25-(OH)D had a significant increase in MTB-induced IFN-γ production after taking vitamin D supplements. Vitamin D administration in large amounts can hasten the recovery of TB patients. The findings point is a therapeutically useful activity of Vitamin D's in the management for tuberculosis.Keywords: tuberculosis, vitamin D, interferon gamma, protein, infection
Procedia PDF Downloads 52844 Formulation, Acceptability, and Characteristics of Instant Surabi Based on Composite Rice-Soybean Flour and Supplemented with Torbangun Powder for Attention Deficit Hyperactivity Disorder Children
Authors: Dewi Hapsari Ratna Muninggar, M. Rizal Martua Damanik
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The purpose of this study was to develop a formulation of instant Indonesian traditional pancake (Surabi) based on composite rice and soybean flour and supplemented with Torbangun (Coleus amboinicus Lour) powder as an alternative snack for ADHD (Attention Deficit Hyperactivity Disorder) children. Completely randomised factorial design by two factors which were the ratio of composite rice and soybean flour (75:25; 70:30; 65:35) as well as the addition of Torbangun powder (3%; 5%; 7%) was used in this study. This study revealed that the best formula was instant surabi with 65:35 composite rice and soybean flour and 5% addition of Torbangun powder by considering hedonic test result, functional aspect and nutrients contribution. Then, both chemical and physical characteristics from the best formula of instant surabi were measured. Nutrients content of the chosen instant surabi per 100 g wet basis were 62.68 g moisture, 1.30 g ash, 6.81 g protein, 0.75 g fat, 28.47 g carbohydrate, 88.62 mg calcium, 4.14 mg iron, and 144 kcal energy while physical characteristics, such as water activity, cohesiveness, and hardness were 0.97, 0.569, 5582.2 g force consecutively. The results of this research suggested that instant surabi which can be possibly beneficial for ADHD children had 65:35 for rice and soybean flour ratio as well as 5% for the addition of Torbangun powder.Keywords: ADHD children, instant surabi, soybean, torbangun
Procedia PDF Downloads 150843 Condition Optimization for Trypsin and Chymotrypsin Activities in Economic Animals
Authors: Mallika Supa-Aksorn, Buaream Maneewan, Jiraporn Rojtinnakorn
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For animals, trypsin and chymotrypsin are the 2 proteases that play the important role in protein digestion and involving in growth rate. In many animals, these two enzymes are indicated as growth parameter by feed. Although enzyme assay at optimal condition is significant for its accuracy activity determination. There is less report of trypsin and chymotrypsin. Therefore, in this study, optimization of pH and temperature for trypsin (T) and chymotrypsin (C) in economic species; i.e. Nile tilapia (Oreochromis niloticus), sand goby (Oxyeleotoris marmoratus), giant freshwater prawn (Macrobachium rosenberchii) and native chicken (Gallus gallus) were investigated. Each enzyme of each species was assaying for its specific activity with variation of pH in range of 2-12 and temperature in range of 30-80 °C. It revealed that, for Nile tilapia, T had optimal condition at pH 9 and temperature 50-80 °C, whereas C had optimal condition at pH 8 and temperature 60 °C. For sand goby, T had optimal condition at pH 7 and temperature of 50 °C, while C had optimal condition at pH 11 and temperature of 70-75 °C. For juvenile freshwater prawn, T had optimal condition at pH 10-11 and temperature of 60-65 °C, C had optimal condition at pH 8 and temperature of 70°C. For starter native chicken, T has optimal condition at pH 7 and temperature of 70 °C, whereas C had o optimal condition at pH 8 and temperature of 60°C. This information of optimal conditions will be high valuable in further for, actual enzyme measurement of T and C activities that benefit for growth and feed analysis.Keywords: trypsin, chymotrypsin, Oreochromis niloticus, Oxyeleotoris marmoratus, Macrobachium rosenberchii, Gallus gallus
Procedia PDF Downloads 259842 Evaluation of Hancornia speciosa Gomes Lyophilization at Different Stages of Maturation
Authors: D. C. Soares, J. T. S. Santos, D. G. Costa, A. K. S. Abud, T. P. Nunes, A. V. D. Figueiredo, A. M. de Oliveira Junior
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Mangabeira (Hancornia speciosa Gomes), a native plant in Brazil, is found growing spontaneously in various regions of the country. The high perishability of tropical fruits such as mangaba, causes it to be necessary to use technologies that promote conservation, aiming to increase the shelf life of this fruit and add value. The objective of this study was to compare the mangabas lyophilisation curves behaviours with different sizes and maturation stages. The fruits were freeze-dried for a period of approximately 45 hours at lyophilizer Liotop brand, model L -108. It has been considered large the fruits between 38 and 58 mm diameter and small, between 23 and 28 mm diameter and the two states of maturation, intermediate and mature. Large size mangabas drying curves in both states of maturation were linear behaviour at all process, while the kinetic drying curves related to small fruits, independent of maturation state, had a typical behaviour of drying, with all the well-defined steps. With these results it was noted that the time of lyophilisation was suitable for small mangabas, a fact that did not happen with the larger one. This may indicate that the large mangabas require a longer time to freeze until reaches the equilibrium level, as it happens with the small fruits, going to have constant moisture at the end of the process. For both types of fruit were analysed water activity, acidity, protein, lipid, and vitamin C before and after the process.Keywords: freeze dryer, mangaba, conservation, chemical characteristics
Procedia PDF Downloads 300841 Differential Proteomic Profile and Terpenoid Production in Somatic Embryos of Jatropha curcas
Authors: Anamarel Medina-Hernandez, Teresa Ponce-Noyola, Ileana Vera-Reyes, Ana C. Ramos-Valdivia
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Somatic embryos reproduce original seed characteristics and could be implemented in biotechnological studies. Jatropha curcas L. is an important plant for biodiesel production, but also is used in traditional medicine. Seeds from J. curcas are toxic because contain diterpenoids called phorbol esters, but in Mexico exist a non-toxic variety. Therefore, somatic embryos suspension cultures from non-toxic J. curcas variety were induced. In order to investigate the characteristics of somatic embryos, a differential proteomic analysis was made between pre-globular and globular stages by 2-D gel electrophoresis. 108 spots were differentially expressed (p<0.02), and 20 spots from globular somatic embryos were sequenced by MALDI-TOF-TOF mass spectrometry. A comparative analysis of terpenoids production between the two stages was made by RP-18 TLC plates. The sequenced proteins were related to energy production (68%), protein destination and storage (9%), secondary metabolism (9%), signal transduction (5%), cell structure (5%) and aminoacid metabolism (4%). Regarding terpenoid production, in pre-globular and globular somatic embryos were identified sterols and triterpenes of pharmacological interest (alpha-amyrin and betulinic acid) but also it was found compounds that were unique to each stage. The results of this work are the basis to characterize at different levels the J. curcas somatic embryos so that this system can be used efficiently in biotechnological processes.Keywords: Jatropha curcas, proteomics, somatic embryo, terpenoids
Procedia PDF Downloads 256840 Novel Pyrimidine Based Semicarbazones: Confirmation of Four Binding Site Pharmacophoric Model Hypothesis for Antiepileptic Activity
Authors: Harish Rajak, Swati Singh
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A series of novel pyrimidine based semicarbazone were designed and synthesized on the basis of semicarbazone based pharmacophoric model to satisfy the structural prerequisite crucial for antiepileptic activity. The semicarbazones based pharmacophoric model consists of following four essential binding sites: (i) An aryl hydrophobic binding site with halo substituent; (ii) A hydrogen bonding domain; (iii) An electron donor group and (iv) Another hydrophobic-hydrophilic site controlling the pharmacokinetic features of the anticonvulsant. The aryl semicarbazones has been recognized as a structurally novel class of compounds with remarkable anticonvulsant activity. In the present study, all the test semicarbazones were subjected to molecular docking using Glide v5.8. Some of the compounds were found to interact with ARG192, GLU270 and THR353 residues of 1OHV protein, present in GABA-AT receptor. The chemical structures of the synthesized molecules were characterized by elemental and spectral (IR, 1H NMR, 13C NMR and MS) analysis. The anticonvulsant activities of the compounds were investigated using maximal electroshock seizure (MES) and subcutaneous pentylenetrtrazole (scPTZ) models. The neurotoxicity was evaluated in mice by the rotorod test. The attempts were also made to establish structure-activity relationships among synthesized compounds. The results of the present study confirmed that the pharmacophore model with four binding sites is essential for antiepileptic activity.Keywords: pyrimidine, semicarbazones, anticonvulsant activity, neurotoxicity
Procedia PDF Downloads 253