Search results for: protein isolates
1729 Functional Characteristics of Chemosensory Proteins in the Sawyer Beetle Monochamus alternatus Hope
Authors: Saqib Ali, Man-Qun Wang
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The Japanese pine sawyer, Monochamus alternatus Hope (Coleoptera: Cerambycidae), is a major pest of pines and it is also the key vector of the exotic pinewood nematode in China. In the present study, we cloned, expressed, and purified a chemosensory protein (CSP) in M. alternatus. We surveyed its expression in various developmental stages of male and female adult tissues and determined its binding affinities for different pine volatiles using a competitive binding fluorescence assay. A CSP known as CSP5 in M. alternatus was obtained from an antennal cDNA library and expressed in Escherichia coli. Quantitative reverse transcription polymerase chain reaction results indicated that the CSP5 gene was mainly expressed in male and female antennae. Competitive binding assays were performed to test the binding affinity of recombinant CSP5 to 13 odour molecules of pine volatiles. The results showed that CSP5 showed very strong binding abilities to myrcene, (+)-β-pinene, and (−)-isolongifolene, whereas the volatiles 2-methoxy-4-vinylphenol, p-cymene, and (+)-limonene oxide have relatively weak binding affinity at pH 5.0. Three volatiles myrcene, (+)-β-pinene, and (−)-isolongifolene may play crucial roles in CSP5 binding with ligands, but this needs further study for confirmation. The sensitivity of insect to host plant volatiles can effectively be used to control and monitor the population through mass trapping as part of integrated pest management programs.Keywords: olfactory-specific protein, volatiles, competitive binding assay, expression characteristics, qPCR
Procedia PDF Downloads 1291728 Activity Anti-Motility Exstract Kedondong Leaf in Balb/C Strain Male Mice Invivo
Authors: Muhammad Abdul Latif, Edijanti Goenarwo , Intan Rahmania Eka
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Diarrhea is one of the leading causes of morbidity and mortality in many countries, as well as responsible for the deaths of millions of people each year. Previous research showed that the leaves, bark, and root bark of kedondong contains saponins, tannins, and flavonoids. Tannins have anti-diarrheal effects that work as the freeze of protein / astrigen, and may inhibit the secretion of chloride over the tannate bonding between protein in the intestines. Chemical compounds of flavonoids also have an effect as anti-diarrheal block receptors Cl ˉ in intestinal thus reducing the secretion of Cl ˉ to the intestinal lume. This research aims to know the anti-diarrheal activity of extracts kedondong leaf in mice Balb/C strain males in vivo. This research also proves kedondong leaves as an anti-diarrhea through trial efficacy of kedondong leaves as antisekretori and antimotilitas. This research using post-test only controlled group design. Analysis of statistical data normality and homogenity were tested by Kolmogorov Smirnov. If the data obtained homogenous then using ANOVA test. This research using ethanolic extracts kedondong leaf 200, 400 and 800 mg/kg BW to prove there is anti-motility became five treatment groups. The result showed dose of ethanolic extracts kedondong leaf 800 mg/kg BW have significant value (p < 0.005). The conclusion from this extracts kedondong leaf research 800 mg/kg BW have pharmacological effects as antimotility on Balb/C strain male mice.Keywords: anti-diarrhea, anti-motility, castrol oil, kedondong leaf
Procedia PDF Downloads 4761727 Constitutive Flo1p Expression on Strains Bearing Deletions in Genes Involved in Cell Wall Biogenesis
Authors: Lethukuthula Ngobese, Abin Gupthar, Patrick Govender
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The ability of yeast cell wall-derived mannoproteins (glycoproteins) to positively contribute to oenological properties has been a key factor that stimulates research initiatives into these industrially important glycoproteins. In addition, and from a fundamental research perspective, yeast cell wall glycoproteins are involved in a wide range of biological interactions. To date, and to the best of our knowledge, our understanding of the fine molecular structure of these mannoproteins is fairly limited. Generally, the amino acid sequences of their protein moieties have been established from structural and functional analysis of the genomic sequence of these yeasts whilst far less information is available on the glycosyl moieties of these mannoproteins. A novel strategy was devised in this study that entails the genetic engineering of yeast strains that over-express and release cell wall-associated glycoproteins into the liquid growth medium. To this end, the Flo1p mannoprotein was overexpressed in Saccharomyces cerevisiae laboratory strains bearing a specific deletion in KNR4 and GPI7 genes involved in cell wall biosynthesis that have been previously shown to extracellularly hyper-secrete cell wall-associated glycoproteins. A polymerase chain reaction (PCR) -based cloning strategy was employed to generate transgenic yeast strains in which the native cell wall FLO1 glycoprotein-encoding gene is brought under transcriptional control of the constitutive PGK1 promoter. The modified Helm’s flocculation assay was employed to assess flocculation intensities of a Flo1p over-expressing wild type and deletion mutant as an indirect measure of their abilities to release the desired mannoprotein. The flocculation intensities of the transformed strains were assessed and all the strains showed similar intensities (>98% flocculation). To assess if mannoproteins were released into the growth medium, the supernatant of each strain was subjected to the BCA protein assay and the transformed Δknr4 strain showed a considerable increase in protein levels. This study has the potential to produce mannoproteins in sufficient quantities that may be employed in future investigations to understand their molecular structures and mechanisms of interaction to the benefit of both fundamental and industrial applications.Keywords: glycoproteins, genetic engineering, flocculation, over-expression
Procedia PDF Downloads 4161726 In vivo Estimation of Mutation Rate of the Aleutian Mink Disease Virus
Authors: P.P. Rupasinghe, A.H. Farid
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The Aleutian mink disease virus (AMDV, Carnivore amdoparvovirus 1) causes persistent infection, plasmacytosis, and formation and deposition of immune complexes in various organs in adult mink, leading to glomerulonephritis, arteritis and sometimes death. The disease has no cure nor an effective vaccine, and identification and culling of mink positive for anti-AMDV antibodies have not been successful in controlling the infection in many countries. The failure to eradicate the virus from infected farms may be caused by keeping false-negative individuals on the farm, virus transmission from wild animals, or neighboring farms. The identification of sources of infection, which can be performed by comparing viral sequences, is important in the success of viral eradication programs. High mutation rates could cause inaccuracies when viral sequences are used to trace back an infection to its origin. There is no published information on the mutation rate of AMDV either in vivo or in vitro. The in vivo estimation is the most accurate method, but it is difficult to perform because of the inherent technical complexities, namely infecting live animals, the unknown numbers of viral generations (i.e., infection cycles), the removal of deleterious mutations over time and genetic drift. The objective of this study was to determine the mutation rate of AMDV on which no information was available. A homogenate was prepared from the spleen of one naturally infected American mink (Neovison vison) from Nova Scotia, Canada (parental template). The near full-length genome of this isolate (91.6%, 4,143 bp) was bidirectionally sequenced. A group of black mink was inoculated with this homogenate (descendant mink). Spleen sampled were collected from 10 descendant mink after 16 weeks post-inoculation (wpi) and from anther 10 mink after 176 wpi, and their near-full length genomes were bi-directionally sequenced. Sequences of these mink were compared with each other and with the sequence of the parental template. The number of nucleotide substitutions at 176 wpi was 3.1 times greater than that at 16 wpi (113 vs 36) whereas the estimates of mutation rate at 176 wpi was 3.1 times lower than that at 176 wpi (2.85×10-3 vs 9.13×10-4 substitutions/ site/ year), showing a decreasing trend in the mutation rate per unit of time. Although there is no report on in vivo estimate of the mutation rate of DNA viruses in animals using the same method which was used in the current study, these estimates are at the higher range of reported values for DNA viruses determined by various techniques. These high estimates are logical based on the wide range of diversity and pathogenicity of AMDV isolates. The results suggest that increases in the number of nucleotide substitutions over time and subsequent divergence make it difficult to accurately trace back AMDV isolates to their origin when several years elapsed between the two samplings.Keywords: Aleutian mink disease virus, American mink, mutation rate, nucleotide substitution
Procedia PDF Downloads 1261725 Clara Cell Secretory Protein 16 Serum Level Decreases in Patients with Non-Smoking-Related Chronic Obstructive Pulmonary Diseases (COPD)
Authors: Lian Wu, Mervyn Merrilees
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Chronic Obstructive Pulmonary Disease (COPD) is a worldwide problem, characterized by irreversible and progressive airflow obstruction. In New Zealand, it is currently the 4th commonest cause of death and exacerbations of COPD are a frequent cause of admission to hospital. Serum levels of Clara cell secretory protein-16 (CC-16) are believed to represent Clara cell toxicity. More recently, CC-16 has been found to be associated with smoker COPD. It is produced almost exclusively by non-ciliated Clara cells in the airways, and its primary function is to protect the lungs against oxidative stress and carcinogenesis. After acute exposure to cigarette smoke, serum levels of CC-16 become elevated. CC16 is a potent natural immune-suppressor and anti-inflammatory agent. In vitro, CC16 inhibits both monocyte and polymorphonuclear neutrophils chemotaxis and phagocytosis. CC16 also inhibits fibroblast chemotaxis. However, the role of CC-16 in non-smoking related COPD is still not clear. In this study, we investigated serum CC-16 levels in non-smoking related COPD. Methods: We compared non-smoker patients with COPD (FEV1<60% of predicted, FEV1/FVC <0.7, n=100) and individuals with normal lung function FEV1≥ 80% of predicted and FEV1/FVC≥ 0.7, n=80). All subjects had no smoking history. CC-16 was measured by ELISA. Results and conclusion: Serum CC-16 levels are reduced in individuals with non-smoking related COPD, and there is a weak correlation with disease severity in non-smoking related COPD group compared to non-smoker controls.Keywords: COPD, CC-16, ELISA, non-smoking-related COPD
Procedia PDF Downloads 3811724 Estimation of Genetic Diversity in Sorghum Accessions Using Agro-Mophological and Nutritional Traits
Authors: Maletsema Alina Mofokeng, Nemera Shargie
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Sorghum is one of the most important cereal crops grown as a source of calories for many people in tropics and sub-tropics of the world. Proper characterisation and evaluation of crop germplasm is an important component for effective management of genetic resources and their utilisation in the improvement of the crop through plant breeding. The objective of the study was to estimate the genetic diversity present in sorghum accessions grown in South Africa using agro-morphological traits and some nutritional contents. The experiment was carried out in Potchefstroom. Data were subjected to correlations, principal components analysis, and hierarchical clustering using GenStat statistical software. There were highly significance differences among the accessions based on agro-morphological and nutritional quality traits. Grain yield was highly positively correlated with panicle weight. Plant height was highly significantly correlated with internode length, leaf length, leaf number, stem diameter, the number of nodes and starch content. The Principal component analysis revealed three most important PCs with a total variation of 78.6%. The protein content ranged from 7.7 to 14.7%, and starch ranged from 58.52 to 80.44%. The accessions that had high protein and starch content were AS16cyc and MP4277. There was vast genetic diversity observed among the accessions assessed that can be used by plant breeders to improve yield and nutritional traits.Keywords: accessions, genetic diversity, nutritional quality, sorghum
Procedia PDF Downloads 2631723 Morphological and Molecular Abnormalities of the Skeletal Muscle Tissue from Pediatric Patient Affected by a Rare Genetic Chaperonopathy Associated with Motor Neuropathy
Authors: Leila Noori, Rosario Barone, Francesca Rappa, Antonella Marino Gammazza, Alessandra Maria Vitale, Giuseppe Donato Mangano, Giusy Sentiero, Filippo Macaluso, Kathryn H. Myburgh, Francesco Cappello, Federica Scalia
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The neuromuscular system controls, directs, and allows movement of the body through the action of neural circuits, which include motor neurons, sensory neurons, and skeletal muscle fibers. Protein homeostasis of the involved cytotypes appears crucial to maintain the correct and prolonged functions of the neuromuscular system, and both neuronal cells and skeletal muscle fibers express significant quantities of protein chaperones, the molecular machinery responsible to maintain the protein turnover. Genetic mutations or defective post-translational modifications of molecular chaperones (i.e., genetic or acquired chaperonopathies) may lead to neuromuscular disorders called as neurochaperonopathies. The limited knowledge of the effects of the defective chaperones on skeletal muscle fibers and neurons impedes the progression of therapeutic approaches. A distinct genetic variation of CCT5 gene encoding for the subunit 5 of the chaperonin CCT (Chaperonin Containing TCP1; also known as TRiC, TCP1 Ring Complex) was recently described associated with severe distal motor neuropathy by our team. In this study, we investigated the histopathological abnormalities of the skeletal muscle biopsy of the pediatric patient affected by the mutation Leu224Val in the CCT5 subunit. We provide molecular and structural features of the diseased skeletal muscle tissue that we believe may be useful to identify undiagnosed cases of this rare genetic disorder. We investigated the histological abnormalities of the affected tissue via hematoxylin and eosin staining. Then we used immunofluorescence and qPCR techniques to explore the expression and distribution of CCT5 in diseased and healthy skeletal muscle tissue. Immunofluorescence and immunohistochemistry assays were performed to study the sarcomeric and structural proteins of skeletal muscle, including actin, myosin, tubulin, troponin-T, telethonin, and titin. We performed Western blot to examine the protein expression of CCT5 and some heat shock proteins, Hsp90, Hsp60, Hsp27, and α-B crystallin, along with the main client proteins of the CCT5, actin, and tubulin. Our findings revealed muscular atrophy, abnormal morphology, and different sizes of muscle fibers in affected tissue. The swollen nuclei and wide interfiber spaces were seen. Expression of CCT5 had been decreased and showed a different distribution pattern in the affected tissue. Altered expression, distribution, and bandage pattern were detected by confocal microscopy for the interested muscular proteins in tissue from the patient compared to the healthy control. Protein levels of the studied Hsps normally located at the Z-disk were reduced. Western blot results showed increased levels of the actin and tubulin proteins in the diseased skeletal muscle biopsy compared to healthy tissue. Chaperones must be expressed at high levels in skeletal muscle to counteract various stressors such as mechanical, oxidative, and thermal crises; therefore, it seems relevant that defects of molecular chaperones may result in damaged skeletal muscle fibers. So far, several chaperones or cochaperones involved in neuromuscular disorders have been defined. Our study shows that alteration of the CCT5 subunit is associated with the damaged structure of skeletal muscle fibers and alterations of chaperone system components and paves the way to explore possible alternative substrates of chaperonin CCT. However, further studies are underway to investigate the CCT mechanisms of action to design applicable therapeutic strategies.Keywords: molecular chaperones, neurochaperonopathy, neuromuscular system, protein homeostasis
Procedia PDF Downloads 711722 Genetic Advance versus Environmental Impact toward Sustainable Protein, Wet Gluten and Zeleny Sedimentation in Bread and Durum Wheat
Authors: Gordana Branković, Dejan Dodig, Vesna Pajić, Vesna Kandić, Desimir Knežević, Nenad Đurić
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The wheat grain quality properties are influenced by genotype, environmental conditions and genotype × environment interaction (GEI). The increasing request of more nutritious wheat products will direct future breeding programmes. Therefore, the aim of investigation was to determine: i) variability of the protein content (PC), wet gluten content (WG) and Zeleny sedimentation volume (ZS); ii) components of variance, heritability in a broad sense (hb2), and expected genetic advance as percent of mean (GAM) for PC, WG, and ZS; iii) correlations between PC, WG, ZS, and most important agronomic traits; in order to assess expected breeding success versus environmental impact for these quality traits. The plant material consisted of 30 genotypes of bread wheat (Triticum aestivum L. ssp. aestivum) and durum wheat (Triticum durum Desf.). The trials were sown at the three test locations in Serbia: Rimski Šančevi, Zemun Polje and Padinska Skela during 2010-2011 and 2011-2012. The experiments were set as randomized complete block design with four replications. The plot consisted of five rows of 1 m2 (5 × 0.2 m × 1 m). PC, WG and ZS were determined by the use of Near infrared spectrometry (NIRS) with the Infraneo analyser (Chopin Technologies, France). PC, WG and ZS, in bread wheat, were in the range 13.4-16.4%, 22.8-30.3%, and 39.4-67.1 mL, respectively, and in durum wheat, in the range 15.3-18.1%, 28.9-36.3%, 37.4-48.3 mL, respectively. The dominant component of variance for PC, WG, and ZS, in bread wheat, was genotype with the genetic variance/GEI variance (VG/VG × E) relation of 3.2, 2.9 and 1.0, respectively, and in durum wheat was GEI with the VG/VG × E relation of 0.70, 0.69 and 0.49, respectively. hb2 and GAM values for PC, WG and ZS, in bread wheat, were 94.9% and 12.6%, 93.7% and 18.4%, and 86.2% and 28.1%, respectively, and in durum wheat, 80.7% and 7.6%, 79.7% and 10.2%, and 74% and 11.2%, respectively. The most consistent through six environments, statistically significant correlations, for bread wheat, were between PC and spike length (-0.312 to -0.637); PC, WG, ZS and grain number per spike (-0.320 to -0.620; -0.369 to -0.567; -0.301 to -0.378, respectively); PC and grain thickness (0.338 to 0.566), and for durum wheat, were between PC, WG, ZS and yield (-0.290 to -0.690; -0.433 to -0.753; -0.297 to -0.660, respectively); PC and plant height (-0.314 to -0.521); PC, WG and spike length (-0.298 to -0.597; -0.293 to -0.627, respectively); PC, WG and grain thickness (0.260 to 0.575; 0.269 to 0.498, respectively); PC, WG and grain vitreousness (0.278 to 0.665; 0.357 to 0.690, respectively). Breeding success can be anticipated for ZS in bread wheat due to coupled high values for hb2 and GAM, suggesting existence of additive genetic effects, and also for WG in bread wheat, due to very high hb2 and medium high GAM. The small, and medium, negative correlations between PC, WG, ZS, and yield or yield components, indicate difficulties to select simultaneously for high quality and yield, depending on linkage for particular genetic arrangements to be broken by recombination.Keywords: bread and durum wheat, genetic advance, protein and wet gluten content, Zeleny sedimentation volume
Procedia PDF Downloads 2541721 Design and Preliminary Evaluation of Benzoxazolone-Based Agents for Targeting Mitochondrial-Located Translocator Protein
Authors: Nidhi Chadha, A. K. Tiwari, Marilyn D. Milton, Anil K. Mishra
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Translocator protein (18 kDa) TSPO is highly expressed during microglia activation in neuroinflammation. Although a number of PET ligands have been developed for the visualization of activated microglia, one of the advantageous approaches is to develop potential optical imaging (OI) probe. Our study involves computational screening, synthesis and evaluation of TSPO ligand through various imaging modalities namely PET/SPECT/Optical. The initial computational screening involves pharmacophore modeling from the library designing having oxo-benzooxazol-3-yl-N-phenyl-acetamide groups and synthesis for visualization of efficacy of these compounds as multimodal imaging probes. Structure modeling of monomer, Ala147Thr mutated, parallel and anti-parallel TSPO dimers was performed and docking analysis was performed for distinct binding sites. Computational analysis showed pattern of variable binding profile of known diagnostic ligands and NBMP via interactions with conserved residues along with TSPO’s natural polymorphism of Ala147→Thr, which showed alteration in the binding affinity due to considerable changes in tertiary structure. Preliminary in vitro binding studies shows binding affinity in the range of 1-5 nm and selectivity was also certified by blocking studies. In summary, this skeleton was found to be potential probe for TSPO imaging due to ease in synthesis, appropriate lipophilicity and reach to specific region of brain.Keywords: TSPO, molecular modeling, imaging, docking
Procedia PDF Downloads 4621720 Magnetic Field Effects on Seed Germination of Phaseolus Vulgaris, Early Seedling Growth, and Chemical Composition
Authors: Farzad Tofigh, Saeideh Najafi, Reza Heidari, Rashid Jamei
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In order to study the effects of magnetic field on the root system and growth of Phaseolus vulgaris, an experiment was conducted in 2012. The possible involvement of magnetic field (MF) pretreatment in physiological factors of Phaseolus vulgaris was investigated. Seeds were subjected to 10 days with 1.8 mT of magnetic field for 1h per day. MF pretreatment decreased the plant height, fresh and dry weight, length of root and length of shoot, Chlorophyll a, Chlorophyll b and carotenoid in 10 days old seedling. In addition, activity of enzymes such as Catalase and Guaiacol peroxidase was decreased due to MF exposure. Also, the total Protein and DPPH content of the treated by magnetic field was not significantly changed in compare to control groups, while the flavonoid, Phenol and prolin content of the treated of the treated by magnetic field was significantly changed in compare to control groups. Lateral branches of roots and secondary roots increased with MF. The results suggest that pretreatment of this MF plays important roles in changes in crop productivity. In all cases there was observed a slight stimulating effect of the factors examined. The growth dynamics were weakened. The plants were shorter. Moreover, the effect of a magnetic field on the crop of Phaseolus vulgaris and its structure was small.Keywords: carotenoid, chlorophyll a, chlorophyll b, DPPH, enzymes, flavonoid, germination, growth, phenol, proline, protein, magnetic field
Procedia PDF Downloads 5041719 In-Situ Fabrication of ZnO PES Membranes for Treatment of Pharmaceuticals
Authors: Oranso T. Mahlangi, Bhekie B. Mamba
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The occurrence of trace organic compounds (TOrCs) in water has raised health concerns for living organisms. The majority of TorCs, including pharmaceuticals and volatile organic compounds, are poorly monitored, partly due to the high cost of analysis and less strict water quality guidelines in South Africa. Therefore, the removal of TorCs is important to guarantee safe potable water. In this study, ZnO nanoparticles were fabricated in situ in polyethersulfone (PES) polymer solutions. This was followed by membrane synthesis using the phase inversion technique. Techniques such as FTIR, Raman, SEM, AFM, EDS, and contact angle measurements were used to characterize the membranes for several physicochemical properties. The membranes were then evaluated for their efficiency in treating pharmaceutical wastewater and resistance to organic (sodium alginate) and protein (bovine serum albumin) fouling. EDS micrographs revealed uniform distribution of ZnO nanoparticles within the polymer matrix, while SEM images showed uniform fingerlike structures. The addition of ZnO increased membrane roughness as well as hydrophilicity (which in turn improved water fluxes). The membranes poorly rejected monovalent and divalent salts (< 10%), making them resistant to flux decline due to concentration polarization effects. However, the membranes effectively removed carbamazepine, caffeine, sulfamethoxazole, ibuprofen, and naproxen by over 50%. ZnO PES membranes were resistant to organic and protein fouling compared to the neat membrane. ZnO PES ultrafiltration membranes may provide a solution in the reclamation of wastewater.Keywords: trace organic compounds, pharmaceuticals, membrane fouling, wastewater reclamation
Procedia PDF Downloads 1411718 Potential Biosorption of Rhodococcus erythropolis, an Isolated Strain from Sossego Copper Mine, Brazil
Authors: Marcela dos P. G. Baltazar, Louise H. Gracioso, Luciana J. Gimenes, Bruno Karolski, Ingrid Avanzi, Elen A. Perpetuo
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In this work, bacterial strains were isolated from environmental samples from a copper mine and three of them presented potential for bioremediation of copper. All the strains were identified by mass spectrometry (MALDI-TOF-Biotyper) and grown in three diferent media supplemented with 100 ppm of copper chloride in flasks of 500mL and it was incubated at 28 °C and 180 rpm. Periodically, samples were taken and monitored for cellular growth and copper biosorption by spectrophotometer UV-Vis (600 nm) and Inductively Coupled Plasma Optical Emission Spectrometry (ICP-OES), respectively. At the end of exponential phase of cellular growth, the biomass was utilized to construct a correlation curve between absorbance and dry mass of the cells. Among the three isolates with potential for biorremediation, 1 strain exhibit capacity the most for bioremediation of effluents contaminated by copper being identified as Rhodococcus erythropolis.Keywords: bioprocess, bioremediation, biosorption, copper
Procedia PDF Downloads 3881717 Study of Germs Responsible of Nosocomial Infections in Hospital of Guelma
Authors: Wissem Abdaoui, Ilhem Mokhtari, Adel Gouri, Benouareth Djamel Eddine
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Contracted in a health facility, hospital-acquired infections are a major public health problem in recent years. The increase of nosocomial infections is partly related to diagnostic and therapeutic advances in medicine. The aim of our study was to isolate and diagnose some types of bacteria that are circulating in the hospital by performing different samples at two medical services: Pulmonary and Infectious Diseases. The antibiotic susceptibility tests were performed for bacterial isolates. The results have shown that there is a predominance of enterobacteria followed by the staphylococcus with its two species epidermidis ans saprophyticus. The study of the antibiogramme identified that some of these bacteria have a resistant profile against all the tested antibiotics. The fight against nosocomial infections is difficult because it must act on several factors: quality of care, safety of the hospital environment, hygiene, wearing gloves etc. are all areas that should be of heightened vigilance and preventive measures.Keywords: nosocomial infection, isolation, identification, sensitivity and resistance to antibiotics
Procedia PDF Downloads 3811716 Molecular Characterization, Host Plant Resistance and Epidemiology of Bean Common Mosaic Virus Infecting Cowpea (Vigna unguiculata L. Walp)
Authors: N. Manjunatha, K. T. Rangswamy, N. Nagaraju, H. A. Prameela, P. Rudraswamy, M. Krishnareddy
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The identification of virus in cowpea especially potyviruses is confusing. Even though there are several studies on viruses causing diseases in cowpea, difficult to distinguish based on symptoms and serological detection. The differentiation of potyviruses considering as a constraint, the present study is initiated for molecular characterization, host plant resistance and epidemiology of the BCMV infecting cowpea. The etiological agent causing cowpea mosaic was identified as Bean Common Mosaic Virus (BCMV) on the basis of RT-PCR and electron microscopy. An approximately 750bp PCR product corresponding to coat protein (CP) region of the virus and the presence of long flexuous filamentous particles measuring about 952 nm in size typical to genus potyvirus were observed under electron microscope. The characterized virus isolate genome had 10054 nucleotides, excluding the 3’ terminal poly (A) tail. Comparison of polyprotein of the virus with other potyviruses showed similar genome organization with 9 cleavage sites resulted in 10 functional proteins. The pairwise sequence comparison of individual genes, P1 showed most divergent, but CP gene was less divergent at nucleotide and amino acid level. A phylogenetic tree constructed based on multiple sequence alignments of the polyprotein nucleotide and amino acid sequences of cowpea BCMV and potyviruses showed virus is closely related to BCMV-HB. Whereas, Soybean variant of china (KJ807806) and NL1 isolate (AY112735) showed 93.8 % (5’UTR) and 94.9 % (3’UTR) homology respectively with other BCMV isolates. This virus transmitted to different leguminous plant species and produced systemic symptoms under greenhouse conditions. Out of 100 cowpea genotypes screened, three genotypes viz., IC 8966, V 5 and IC 202806 showed immune reaction in both field and greenhouse conditions. Single marker analysis (SMA) was revealed out of 4 SSR markers linked to BCMV resistance, M135 marker explains 28.2 % of phenotypic variation (R2) and Polymorphic information content (PIC) value of these markers was ranged from 0.23 to 0.37. The correlation and regression analysis showed rainfall, and minimum temperature had significant negative impact and strong relationship with aphid population, whereas weak correlation was observed with disease incidence. Path coefficient analysis revealed most of the weather parameters exerted their indirect contributions to the aphid population and disease incidence except minimum temperature. This study helps to identify specific gaps in knowledge for researchers who may wish to further analyse the science behind complex interactions between vector-virus and host in relation to the environment. The resistant genotypes identified are could be effectively used in resistance breeding programme.Keywords: cowpea, epidemiology, genotypes, virus
Procedia PDF Downloads 2371715 Preventive Effect of Zinc on Nickel Hepatotoxicity and Nephrotoxicity in Albino (Wistar) Rats
Authors: Zine Kechrid, Samira Bouhalit
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Aim: We studied the effect of intraperitonial zinc treatment on nickel sulphate-induced hepatotoxicity and nephrotoxicity in Wistar strain male albino rats. Materials and Methods: Liver and kidney dysfunction parameters represented by aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), blood glucose, serum total protein, serum urea, serum creatinine, and serum belurebin were estimated. Liver glutathione level, catalase and GPx activities were also determined in liver as indicators of oxidative damage. Result: Nickel treatment led to high serum glucose concentration and produced hepatotoxicity and nephrotoxicity characterized by increasing GPT, GOT and alkaline phosphatase activities, serum total protein, serum urea, serum creatinine and serum belurebin concentrations. In addition, liver glutathione level, catalase and GSH-Px activities diminished due to high lipid peroxidation. The simultaneous administration of zinc with nickel sulphate resulted in a remarkable improvement of the previous parameters compared with rats treated with nickel alone. Conclusion: In conclusion, nickel sulphate led to liver and kidney dysfunctions and hepatic lipid peroxidation in animals, but simultaneous treatment with zinc offers a relative protection against nickel induced hepatotoxicity, nephrotoxicity and lipid peroxidation.Keywords: nickel, zinc, rats, GOT, GPT, nephrotoxicity, hepatotoxicity
Procedia PDF Downloads 4511714 Preparation of Pegylated Interferon Alpha-2b with High Antiviral Activity Using Linear 20 KDa Polyethylene Glycol Derivative
Authors: Ehab El-Dabaa, Omnia Ali, Mohamed Abd El-Hady, Ahmed Osman
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Recombinant human interferon alpha 2 (rhIFN-α2) is FDA approved for treatment of some viral and malignant diseases. Approved pegylated rhIFN-α2 drugs have highly improved pharmacokinetics, pharmacodynamics and therapeutic efficiency compared to native protein. In this work, we studied the pegylation of purified properly refolded rhIFN-α2b using linear 20kDa PEG-NHS (polyethylene glycol- N-hydroxysuccinimidyl ester) to prepare pegylated rhIFN-α2b with high stability and activity. The effect of different parameters like rhIFN-α2b final concentration, pH, rhIFN-α2b/PEG molar ratios and reaction time on the efficiency of pegylation (high percentage of monopegylated rhIFN-α2b) have been studied in small scale (100µl) pegylation reaction trials. Study of the percentages of different components of these reactions (mono, di, polypegylated rhIFN-α2b and unpegylated rhIFN-α2b) indicated that 2h is optimum time to complete the reaction. The pegylation efficiency increased at pH 8 (57.9%) by reducing the protein concentration to 1mg/ml and reducing the rhIFN-α2b/PEG ratio to 1:2. Using larger scale pegylation reaction (65% pegylation efficiency), ion exchange chromatography method has been optimized to prepare and purify the monopegylated rhIFN-α2b with high purity (96%). The prepared monopegylated rhIFN-α2b had apparent Mwt of approximately 65 kDa and high in vitro antiviral activity (2.1x10⁷ ± 0.8 x10⁷ IU/mg). Although it retained approximately 8.4 % of the antiviral activity of the unpegylated rhIFN-α2b, its activity is high compared to other pegylated rhIFN-α2 developed by using similar approach or higher molecular weight branched PEG.Keywords: antiviral activity, rhIFN-α2b, pegylation, pegylation efficiency
Procedia PDF Downloads 1791713 Isolation and Molecular Identification of Two Fungal Strains Capable of Degrading hydrocarbon Contaminants on Saudi Arabian Environment
Authors: Amr A. EL Hanafy, Yasir Anwar, Saleh A. Mohamed, Saleh Mohamed Saleh Al-Garni, Jamal S. M. Sabir , Osama A. H. Abu Zinadah, Mohamed Morsi Ahmed
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In the vicinity of the red sea about 15 fungi species were isolated from oil contaminated sites. On the basis of aptitude to degrade the crude oil and DCPIP assay, two fungal isolates were selected amongst 15 oil degrading strains. Analysis of ITS-1, ITS-2 and amplicon pyrosequencing studies of fungal diversity revealed that these strains belong to Penicillium and Aspergillus species. Two strains that proved to be the most efficient in degrading crude oil was Aspergillus niger (54 %) and Penicillium commune (48 %) Subsequent to two weeks of cultivation in BHS medium the degradation rate were recorded by using spectrophotometer and GC-MS. Hence, it is cleared that these fungal strains has the capability of degradation and can be utilized for cleaning the Saudi Arabian environment.Keywords: fungal strains, hydrocarbon contaminants, molecular identification, biodegradation, GC-MS
Procedia PDF Downloads 5231712 In silico Designing and Insight into Antimalarial Potential of Chalcone-Quinolinylpyrazole Hybrids by Preclinical Study in Mice
Authors: Deepika Saini, Sandeep Jain, Ajay Kumar
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The quinoline scaffold is one of the most widely studied in the discovery of derivatives with various heterocyclic moieties due to its potential antimalarial activities. In the present study, a chalcone series of quinoline derivatives clubbed with pyrazole were synthesized to evaluate their antimalarial property by in vitro schizont maturation inhibition assay against both chloroquine sensitive, 3D7 and chloroquine resistant, RKL9 strain of Plasmodium falciparum. Further, top five compounds were studied for in vivo preclinical study for antimalarial potential against P. berghei in Swiss albino mice. To understand the mechanism of synthesized analogues, they were screened computationally by molecular docking techniques. Compounds were docked into the active site of a protein receptor, Plasmodium falciparum Cysteine Protease Falcipain-2. The compounds were successfully synthesized, and structural confirmation was performed by FTIR, 1H-NMR, mass spectrometry and elemental analysis. In vitro study suggested that the compounds 5b, 5g, 5l, 5s and 5u possessed best antimalarial activity and further tested for in vivo screening. Compound 5u (CH₃ on both rings) with EC₅₀ 0.313 & 0.801 µg/ml against CQ-S & CQ-R strains of P. falciparum respectively and 78.01% suppression of parasitemia. The molecular docking studies of the compounds helped in understanding the mechanism of action against falcipain-2. The present study reveals the binding signatures of the synthesized ligands within the active site of the protein, and it explains the results from in vitro study in their EC₅₀ values and percentage parasitemia.Keywords: antimalarial activity, chalcone, docking, quinoline
Procedia PDF Downloads 4091711 Diversity of Halophilic Archaea from Ezzemoul sabkha in Algeria
Authors: Karima Kharroub, Mohamed Amine Gomri, Mercedes Monteoliva-Sanchez
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Sabkhas and chotts are examples of hypersaline environments inhabited by halophilic microorganisms. In the present study, a culture-dependent methodology was used to determine the archaeal diversity present within Ezzemoul sabkha located in the north-eastern of Algeria. It was assessed using different media with different substrates in attempt to initiate the growth of wide spectrum of halophiles. Several strains were isolated and subjected to a polyphasic taxonomic characterization. The isolates tested placed within the family Halobacteriaceae, a group of extremely halophilic, aerobic archaea that have a salinity tolerance of 3 to 4M and associated with six genera Halorubrum, Halobacterium, Haloterrigena, Haloferax, Halomicrobium and Haloarcula. Halorubrum species were found to be the dominant archaea community as indicated by the presence of different strains recovered from this sabkha. In the other hand, two new species within Halobacteriaceae family; Halorubrum ezzemolulense sp. nov. and Halomicrobium katesii sp. nov. were described.Keywords: sabkha, archaea, halophilic, hypersaline environments
Procedia PDF Downloads 3151710 Nutritional Value and Forage Quality Indicators in Some Rangeland’s Species at Different Vegetation Forms
Authors: Reza Dehghani Bidgoli
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Information on different rangeland plants’ nutritive values at various phonological stages is important in rangelands management. This information helps rangeland managers to choose proper grazing times to achieve higher animal performance without detrimental effects on the rangeland vegetations. Effects of various plant parts’ phonological stages and vegetation types on reserve carbohydrates and forage quality indicators were investigated during the 2009 and 2010. Plant samples were collected in a completely randomized block (CRB) design. The species included, grasses (Secale montanum and Festuco ovina), forbs (Lotus corniculatus and Sanguisorba minor), and shrubs (Kochia prosterata and Salsola rigida). Aerial plant parts’ samples were oven-dried at 80oC for 24 hours, then analyzed for soluble carbohydrates, crude protein (CP), acid detergent fiber (ADF), dry matter digestible (DMD), and metabolizable energy (ME). Results showed that plants at the seedling stage had more reserve carbohydrates and from the three vegetation types (grass, forbs, and shrub), forbs contained more soluble carbohydrates compared to the other two (grasses and shrubs). Differences in soluble carbohydrate contents of different species at various phonological stages in 2 years were statistically significant. The forage quality indicators (CP, ADF, DMD, and ME) in different species, in different vegetation types, in the 2 years were statistically significant, except for the CP.Keywords: grazing, soluble carbohydrate, protein, fiber, metabolizeable energy
Procedia PDF Downloads 2831709 The Proximate Composition and Phytochemical Screening of Momordica Balsamina (Balsam Apple) Fruit and Leaves
Authors: Viruska Jaichand, John Jason Mellem, Viresh Mohanlall
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Malnutrition is a global issue that affects both children and adults, irrespective of their socio-economic status. It is, therefore, important to find various means to tackle malnutrition. This is especially important as undernutrition and overnutrition can be linked to a variety of non-communicable diseases (NCDs). This study aimed to gather more insight into the nutritional and phytochemical quality of Momordica balsamina leaves and fruit (fruit pericarp, fruit flesh and seeds). Results showed that Momordica balsamina had a nutritional composition that would be advantageous to the human diet. The nutritional quality is verified by the presence of a high protein percentage across all samples (19.72%-29.08%), with the leaves containing the highest protein content (29.08%±0.77). There was also a low-fat content present across all samples, which ranged from 1.03% to 2.40%. The ash content indicated the presence of total minerals to be adequate (2.93%-21.16%), where the pericarp had the highest ash quantity (21.16%±0.09). The moisture levels were low (7.11%-13.40%). Momordica balsamina seeds had the highest carbohydrate content (67.84%±0.30). Rich in the major phytoconstituents, Momordica balsamina extracts were found to contain alkaloids, saponins, cardiac glycosides, steroids and triterpenoids. Based on these findings, it can thus be said that the incorporation of Momordica balsamina into an individual’s diet could prevent diseases associated with malnutrition, as well as it could be used to supplement the human diet in managing certain NCDs. Even though there were a number of bioactive compounds detected, further studies which would correlate the phytochemical constituents detected in Momordica balsamina and its effectiveness in treating various diseases are recommended.Keywords: momordica balsamina, nutrients, proximate composition, bioactive compounds, phytoconstituents
Procedia PDF Downloads 751708 Characterization of a Lipolytic Enzyme of Pseudomonas nitroreducens Isolated from Mealworm's Gut
Authors: Jung-En Kuan, Whei-Fen Wu
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In this study, a symbiotic bacteria from yellow mealworm's (Tenebrio molitor) mid-gut was isolated with characteristics of growth on minimal-tributyrin medium. After a PCR-amplification of its 16s rDNA, the resultant nucleotide sequences were then analyzed by schemes of the phylogeny trees. Accordingly, it was designated as Pseudomonas nitroreducens D-01. Next, by searching the lipolytic enzymes in its protein data bank, one of those potential lipolytic α/β hydrolases was identified, again using PCR-amplification and nucleotide-sequencing methods. To construct an expression of this lipolytic gene in plasmids, the target-gene primers were then designed, carrying the C-terminal his-tag sequences. Using the vector pET21a, a recombinant lipolytic hydrolase D gene with his-tag nucleotides was successfully cloned into it, of which the lipolytic D gene is under a control of the T7 promoter. After transformation of the resultant plasmids into Eescherichia coli BL21 (DE3), an IPTG inducer was used for the induction of the recombinant proteins. The protein products were then purified by metal-ion affinity column, and the purified proteins were found capable of forming a clear zone on tributyrin agar plate. Shortly, its enzyme activities were determined by degradation of p-nitrophenyl ester(s), and the substantial yellow end-product, p-nitrophenol, was measured at O.D.405 nm. Specifically, this lipolytic enzyme efficiently targets p-nitrophenyl butyrate. As well, it shows the most reactive activities at 40°C, pH 8 in potassium phosphate buffer. In thermal stability assays, the activities of this enzyme dramatically drop when the temperature is above 50°C. In metal ion assays, MgCl₂ and NH₄Cl induce the enzyme activities while MnSO₄, NiSO₄, CaCl₂, ZnSO₄, CoCl₂, CuSO₄, FeSO₄, and FeCl₃ reduce its activities. Besides, NaCl has no effects on its enzyme activities. Most organic solvents decrease the activities of this enzyme, such as hexane, methanol, ethanol, acetone, isopropanol, chloroform, and ethyl acetate. However, its enzyme activities increase when DMSO exists. All the surfactants like Triton X-100, Tween 80, Tween 20, and Brij35 decrease its lipolytic activities. Using Lineweaver-Burk double reciprocal methods, the function of the enzyme kinetics were determined such as Km = 0.488 (mM), Vmax = 0.0644 (mM/min), and kcat = 3.01x10³ (s⁻¹), as well the total efficiency of kcat/Km is 6.17 x10³ (mM⁻¹/s⁻¹). Afterwards, based on the phylogenetic analyses, this lipolytic protein is classified to type IV lipase by its homologous conserved region in this lipase family.Keywords: enzyme, esterase, lipotic hydrolase, type IV
Procedia PDF Downloads 1331707 Identification of Potential Small Molecule Regulators of PERK Kinase
Authors: Ireneusz Majsterek, Dariusz Pytel, J. Alan Diehl
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PKR-like ER kinase (PERK) is serine/threonie endoplasmic reticulum (ER) transmembrane kinase activated during ER-stress. PERK can activate signaling pathways known as unfolded protein response (UPR). Attenuation of translation is mediated by PERK via phosphorylation of eukaryotic initiation factor 2α (eIF2α), which is necessary for translation initiation. PERK activation also directly contributes to activation of Nrf2 which regulates expression of anti-oxidant enzymes. An increased phosphorylation of eIF2α has been reported in Alzheimer disease (AD) patient hippocampus, indicating that PERK is activated in this disease. Recent data have revealed activation of PERK signaling in non-Hodgkins lymphomas. Results also revealed that loss of PERK limits mammary tumor cell growth in vitro and in vivo. Consistent with these observations, activation of UPR in vitro increases levels of the amyloid precursor protein (APP), the peptide from which beta-amyloid plaques (AB) fragments are derived. Finally, proteolytic processing of APP, including the cleavages that produce AB, largely occurs in the ER, and localization coincident with PERK activity. Thus, we expect that PERK-dependent signaling is critical for progression of many types of diseases (human cancer, neurodegenerative disease and other). Therefore, modulation of PERK activity may be a useful therapeutic target in the treatment of different diseases that fail to respond to traditional chemotherapeutic strategies, including Alzheimer’s disease. Our goal will be to developed therapeutic modalities targeting PERK activity.Keywords: PERK kinase, small molecule inhibitor, neurodegenerative disease, Alzheimer’s disease
Procedia PDF Downloads 4831706 Influence of Protein Malnutrition and Different Stressful Conditions on Aluminum-Induced Neurotoxicity in Rats: Focus on the Possible Protection Using Epigallocatechin-3-Gallate
Authors: Azza A. Ali, Asmaa Abdelaty, Mona G. Khalil, Mona M. Kamal, Karema Abu-Elfotuh
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Background: Aluminium (Al) is known as a neurotoxin environmental pollutant that can cause certain diseases as Dementia, Alzheimer's disease, and Parkinsonism. It is widely used in antacid drugs as well as in food additives and toothpaste. Stresses have been linked to cognitive impairment; Social isolation (SI) may exacerbate memory deficits while protein malnutrition (PM) increases oxidative damage in cortex, hippocampus and cerebellum. The risk of cognitive decline may be lower by maintaining social connections. Epigallocatechin-3-gallate (EGCG) is the most abundant catechin in green tea and has antioxidant, anti-inflammatory and anti-atherogenic effects as well as health-promoting effects in CNS. Objective: To study the influence of different stressful conditions as social isolation, electric shock (EC) and inadequate Nutritional condition as PM on neurotoxicity induced by Al in rats as well as to investigate the possible protective effect of EGCG in these stressful and PM conditions. Methods: Rats were divided into two major groups; protected group which was daily treated during three weeks of the experiment by EGCG (10 mg/kg, IP) or non-treated. Protected and non-protected groups included five subgroups as following: One normal control received saline and four Al toxicity groups injected daily for three weeks by ALCl3 (70 mg/kg, IP). One of them served as Al toxicity model, two groups subjected to different stresses either by isolation as mild stressful condition (SI-associated Al toxicity model) or by electric shock as high stressful condition (EC- associated Al toxicity model). The last was maintained on 10% casein diet (PM -associated Al toxicity model). Isolated rats were housed individually in cages covered with black plastic. Biochemical changes in the brain as acetyl cholinesterase (ACHE), Aβ, brain derived neurotrophic factor (BDNF), inflammatory mediators (TNF-α, IL-1β), oxidative parameters (MDA, SOD, TAC) were estimated for all groups. Histopathological changes in different brain regions were also evaluated. Results: Rats exposed to Al for three weeks showed brain neurotoxicity and neuronal degenerations. Both mild (SI) and high (EC) stressful conditions as well as inadequate nutrition (PM) enhanced Al-induced neurotoxicity and brain neuronal degenerations; the enhancement induced by stresses especially in its higher conditions (ES) was more pronounced than that of inadequate nutritional conditions (PM) as indicated by the significant increase in Aβ, ACHE, MDA, TNF-α, IL-1β together with the significant decrease in SOD, TAC, BDNF. On the other hand, EGCG showed more pronounced protection against hazards of Al in both stressful conditions (SI and EC) rather than in PM .The protective effects of EGCG were indicated by the significant decrease in Aβ, ACHE, MDA, TNF-α, IL-1β together with the increase in SOD, TAC, BDNF and confirmed by brain histopathological examinations. Conclusion: Neurotoxicity and brain neuronal degenerations induced by Al were more severe with stresses than with PM. EGCG can protect against Al-induced brain neuronal degenerations in all conditions. Consequently, administration of EGCG together with socialization as well as adequate protein nutrition is advised especially on excessive Al-exposure to avoid the severity of its neuronal toxicity.Keywords: environmental pollution, aluminum, social isolation, protein malnutrition, neuronal degeneration, epigallocatechin-3-gallate, rats
Procedia PDF Downloads 3911705 A Molecular Modelling Approach for Identification of Lead Compound from Rhizomes of Glycosmis Pentaphylla for Skin Cancer Treatment
Authors: Rahul Shrivastava, Manish Tripathi, Mohmmad Yasir, Shailesh Singh
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Life style changes and depletion in atmospheric ozone layer in recent decades lead to increase in skin cancer including both melanoma and nonmelanomas. Natural products which were obtained from different plant species have the potential of anti skin cancer activity. In regard of this, present study focuses the potential effect of Glycosmis pentaphylla against anti skin cancer activity. Different Phytochemical constituents which were present in the roots of Glycosmis pentaphylla were identified and were used as ligands after sketching of their structures with the help of ACD/Chemsketch. These ligands are screened for their anticancer potential with proteins which are involved in skin cancer effects with the help of pyrx software. After performing docking studies, results reveal that Noracronycine secondary metabolite of Glycosmis pentaphylla shows strong affinity of their binding energy with Ribosomal S6 Kinase 2 (2QR8) protein. Ribosomal S6 Kinase 2 (2QR8) has an important role in the cell proliferation and transformation mediated through by N-terminal kinase domain and was induced by the tumour promoters such as epidermal growth factor. It also plays a key role in the neoplastic transformation of human skin cells and in skin cancer growth. Noracronycine interact with THR-493 and MET-496 residue of Ribosomal S6 Kinase 2 protein with binding energy ΔG = -8.68 kcal/mole. Thus on the basis of this study we can say that Noracronycine which present in roots of Glycosmis pentaphylla can be used as lead compound against skin cancer.Keywords: glycosmis pentaphylla, pyrx, ribosomal s6 kinase, skin cancer
Procedia PDF Downloads 3031704 YHV-Responsive Gene Expression under the Influence of PmRelish Regulation
Authors: Suwattana Visetnan, Premruethai Supungul, Sureerat Tang, Ikuo Hirono, Anchalee Tassanakajon, Vichien Rimphanitchayakit
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In animals, infection by Gram-negative bacteria and certain viruses activates the Imd signaling pathway wherein the a NF-κB transcription factor, Relish, is a key regulatory protein for the synthesis of antimicrobial proteins. Infection by yellow head virus (YHV) activates the Imd pathway. To investigate the expression of genes involved in YHV infection and under the influence of PmRelish regulation, RNA interference and suppression subtractive hybridization (SSH) are employed. The genes in forward library expressed in shrimp after YHV infection and under the activity of PmRelish were obtained by subtracting the cDNAs from YHV-infected and PmRelish-knockdown shrimp with cDNAs from YHV-infected shrimp. Opposite subtraction gave a reverse library whereby an alternative set of genes under YHV infection and no PmRelish expression was obtained. Sequencing of 252 and 99 cDNA clones from the respective forward and reverse libraries were done and annotated through blast search against the GenBank sequences. Genes involved in defense and homeostasis were abundant in both libraries, 31% and 23% in the forward and reverse libraries, respectively. They were predominantly antimicrobial proteins, proteinases and proteinase inhibitors. The expression of antimicrobial protein genes, ALFPm3, crustinPm1, penaeidin3 and penaeidin5 were tested under PmRelish silencing and Gram-negative bacterium V. harveyi infection. Together with the results previously reported, the expression of penaeidin5 and also penaeidin3 but not ALFPm3 and crustinPm1 were under the regulation of PmRelish in the Imd pathway.Keywords: relish, yellow head virus, penaeus monodon, antimicrobial proteins
Procedia PDF Downloads 2121703 Insights Into Serotonin-Receptor Binding and Stability via Molecular Dynamics Simulations: Key Residues for Electrostatic Interactions and Signal Transduction
Authors: Arunima Verma, Padmabati Mondal
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Serotonin-receptor binding plays a key role in several neurological and biological processes, including mood, sleep, hunger, cognition, learning, and memory. In this article, we performed molecular dynamics simulation to examine the key residues that play an essential role in the binding of serotonin to the G-protein-coupled 5-HT₁ᴮ receptor (5-HT₁ᴮ R) via electrostatic interactions. An end-point free energy calculation method (MM-PBSA) determines the stability of the 5-HT1B R due to serotonin binding. The single-point mutation of the polar or charged amino acid residues (Asp129, Thr134) on the binding sites and the calculation of binding free energy validate the importance of these residues in the stability of the serotonin-receptor complex. Principal component analysis indicates the serotonin-bound 5-HT1BR is more stabilized than the apo-receptor in terms of dynamical changes. The difference dynamic cross-correlations map shows the correlation between the transmembrane and mini-Go, which indicates signal transduction happening between mini-Go and the receptor. Allosteric communication reveals the key nodes for signal transduction in 5-HT1BR. These results provide useful insights into the signal transduction pathways and mutagenesis study to regulate the functionality of the complex. The developed protocols can be applied to study local non-covalent interactions and long-range allosteric communications in any protein-ligand system for computer-aided drug design.Keywords: allostery, CADD, MD simulations, MM-PBSA
Procedia PDF Downloads 871702 Probiotics’ Antibacterial Activity on Beef and Camel Minced Meat at Altered Ranges of Temperature
Authors: Rania Samir Zaki
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Because of their inhibitory effects, selected probiotic Lactobacilli may be used as antimicrobial against some hazardous microorganisms responsible for spoilage of fresh minced beef (cattle) minced meat and camel minced meat. Lactic acid bacteria were isolated from camel meat. These included 10 isolates; 1 Lactobacillus fermenti, 4 Lactobacillus plantarum, 4 Lactobacillus pulgaricus, 3 Lactobacillus acidophilus and 1 Lactobacillus brevis. The most efficient inhibitory organism was Lactobacillus plantarum which can be used as a propiotic with antibacterial activity. All microbiological analyses were made at the time 0, first day and the second day at altered ranges of temperature [4±2 ⁰C (chilling temperature), 25±2 ⁰C, and 38±2 ⁰C]. Results showed a significant decrease of pH 6.2 to 5.1 within variant types of meat, in addition to reduction of Total Bacterial Count, Enterococci, Bacillus cereus and Escherichia coli together with the stability of Coliforms and absence of Staphylococcus aureus.Keywords: antibacterial, camel meat, inhibition, probiotics
Procedia PDF Downloads 2991701 Histone Deacetylases Inhibitor - Valproic Acid Sensitizes Human Melanoma Cells for alkylating agent and PARP inhibitor
Authors: Małgorzata Drzewiecka, Tomasz Śliwiński, Maciej Radek
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The inhibition of histone deacetyles (HDACs) holds promise as a potential anti-cancer therapy because histone and non-histone protein acetylation is frequently disrupted in cancer, leading to cancer initiation and progression. Additionally, histone deacetylase inhibitors (HDACi) such as class I HDAC inhibitor - valproic acid (VPA) have been shown to enhance the effectiveness of DNA-damaging factors, such as cisplatin or radiation. In this study, we found that, using of VPA in combination with talazoparib (BMN-637 – PARP1 inhibitor – PARPi) and/or Dacarabazine (DTIC - alkylating agent) resulted in increased DNA double strand break (DSB) and reduced survival (while not affecting primary melanocytes )and proliferation of melanoma cells. Furthermore, pharmacologic inhibition of class I HDACs sensitizes melanoma cells to apoptosis following exposure to DTIC and BMN-637. In addition, inhibition of HDAC caused sensitization of melanoma cells to dacarbazine and BMN-637 in melanoma xenografts in vivo. At the mRNA and protein level histone deacetylase inhibitor downregulated RAD51 and FANCD2. This study provides that combining HDACi, alkylating agent and PARPi could potentially enhance the treatment of melanoma, which is known for being one of the most aggressive malignant tumors. The findings presented here point to a scenario in which HDAC via enhancing the HR-dependent repair of DSBs created during the processing of DNA lesions, are essential nodes in the resistance of malignant melanoma cells to methylating agent-based therapies.Keywords: melanoma, hdac, parp inhibitor, valproic acid
Procedia PDF Downloads 821700 Faults Diagnosis by Thresholding and Decision tree with Neuro-Fuzzy System
Authors: Y. Kourd, D. Lefebvre
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The monitoring of industrial processes is required to ensure operating conditions of industrial systems through automatic detection and isolation of faults. This paper proposes a method of fault diagnosis based on a neuro-fuzzy hybrid structure. This hybrid structure combines the selection of threshold and decision tree. The validation of this method is obtained with the DAMADICS benchmark. In the first phase of the method, a model will be constructed that represents the normal state of the system to fault detection. Signatures of the faults are obtained with residuals analysis and selection of appropriate thresholds. These signatures provide groups of non-separable faults. In the second phase, we build faulty models to see the flaws in the system that cannot be isolated in the first phase. In the latest phase we construct the tree that isolates these faults.Keywords: decision tree, residuals analysis, ANFIS, fault diagnosis
Procedia PDF Downloads 628