Search results for: recombinant protein
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 2414

Search results for: recombinant protein

1574 A Novel Peptide Showing Universal Effect against Multiple Viruses in Vitro and in Vivo

Authors: Hanjun Zhao, Ke Zhang, Bojian Zheng

Abstract:

Background: So far, there is no universal antiviral agent which can inhibit multiple viral infections. More and more drug-resistant viral strains emerge after the antiviral drug application for treatment. Defensins are the front line of host innate immunity and have broad spectrum antibacterial and antiviral effects. However, there is limited data to show if these defensins have good antiviral activity in vivo and what the antiviral mechanism is. Subjects: To investigate a peptide with widespread antivirus activity in vitro and in vivo and illustrate the antiviral mechanism. Methods: Antiviral peptide library designed from mouse beta defensins was synthesized by the company. Recombinant beta defensin was obtained from E. coli. Antiviral activity in vitro was assayed by plaque assay, qPCR. Antiviral activity in vivo was detected by animal challenge with 2009 pandemic H1N1 influenza A virus. The antiviral mechanism was assayed by western blot, ELISA, and qPCR. Conclusions: We identify a new peptide which has widespread effects against multiple viruses (H1N1, H5N1, H7N9, MERS-CoV) in vitro and has efficient antivirus activity in vivo. This peptide inhibits viral entry into target cells and subsequently blocks viral replication. The in vivo study of the antiviral peptide against other viral infections and the investigation of its more detail antiviral mechanism are ongoing.

Keywords: antiviral peptide, defensin, Influenza A virus, mechanism

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1573 Amelioration of Salinity Stress in Spinach (Spinace oleracae) by Exogenous Application of Triacontanol

Authors: Ameer Khan, Iffat Jamal, Ambreen Azam

Abstract:

An experiment was conducted in the Department of Botany, University of Sargodha to observe the amelioration of salinity stress in spinach (Spinacia oleracea) by exogenous application of Triacontanol. Two spinach cultivars (Spinacea oleracea and Rumax dentatus) were obtained from the Agriculture Research institute, Faisalabad. This experiment was conducted in pots. Each pot was filled with 9kg mixture of (sand + soil). Different salinity levels (0mM, 60mM, and 120mM) were created with NaCl according to the saturation percentage of soil after two weeks of seed germination. After the two weeks of salinity treatment, different levels of Triacontanol (0µM, 10µM, 20µM) were applied as foliar spray. Triacontanol was applied along with Tween 80 as surfactant. After the two weeks of Triacontanol application different growth, physiological and biochemical parameters were collected from the experimental study. Both treatments of Triacontanol (10µM, 20µM) were effective to ameliorate the effect of salinity, but 20µM Triacontanol was more effective to increase the shoot length, shoot, root fresh and dry weight. Chlorophyll contents were (chl a, chl b, total chl). Different biochemical parameters were also collected from experimental study. Saline growth medium increased the accumulation of protein and decreased the total free amino acids, and total soluble sugar under salt stress. Application of Triacontanol increased the protein contents. Overall, Application of triacontanol mitigated the effect of salinity.

Keywords: salinity, triacontanol, spinach, biochemical, physiological

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1572 Acute Effects of Exogenous Hormone Treatments on Postprandial Acylation Stimulating Protein Levels in Ovariectomized Rats After a Fat Load

Authors: Bashair Al Riyami

Abstract:

Background: Acylation stimulating protein (ASP) is a small basic protein that was isolated based on its function as a potent lipogenic factor. The role of ASP in lipid metabolism has been described in numerous studies. Several association studies suggest that ASP may play a prominent role in female fat metabolism and distribution. Progesterone is established as a female lipogenic hormone, however the mechanisms by which progesterone exert its effects are not fully understood. AIM: Since ASP is an established potent lipogenic factor with a known mechanism of action, in this study we aim to investigate acute effects of different hormone treatments on ASP levels in vivo after a fat load. Methods: This is a longitudinal study including 24 female wister rats that were randomly divided into 4 groups including controls (n=6). The rats were ovariectomized, and fourteen days later the fasting rats were injected subcutaneously with a single dose of different hormone treatments (progesterone, estrogen and testosterone). An hour later, olive was administered by oral gavage, and plasma blood samples were collected at several time points after oil administration for ASP and triglyceride measurements. Area under the curve (TG-AUC) was calculated to represent TG clearance Results: RM-ANCOVA and post-analysis showed that only the progesterone treated group had a significant postprandial ASP increase at two hours compared to basal levels and to the controls (439.8± 62.4 vs 253.45± 59.03 ug/ml), P= 0.04. Interestingly, increased postprandial ASP levels coordinated negatively with corresponding TG levels and TG-AUC across the postprandial period most apparent in the progesterone and testosterone treated groups that behaved in an opposite manner. ASP levels were 3-fold higher in the progesterone compared to the testosterone treated group, whereas TG-AUC was significantly lower in the progesterone treated group compared to the testosterone treated group. Conclusion: These findings suggest that progesterone treatment enhances ASP production and TG clearance in a simultaneous manner. The strong association of postprandial ASP levels and TG clearance in the progesterone treated group support the notion of a stimulatory role for progesterone on ASP mediated TG clearance. This is the first functional study to demonstrate a cause-effect relationship between hormone treatment and ASP levels in vivo. These findings are promising and may contribute to further understanding the mechanism of progesterone function as a female lipogenic hormone through enhancing ASP production and plasma levels.

Keywords: ASP, lipids, sex hormones, wister rats

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1571 An Inverse Docking Approach for Identifying New Potential Anticancer Targets

Authors: Soujanya Pasumarthi

Abstract:

Inverse docking is a relatively new technique that has been used to identify potential receptor targets of small molecules. Our docking software package MDock is well suited for such an application as it is both computationally efficient, yet simultaneously shows adequate results in binding affinity predictions and enrichment tests. As a validation study, we present the first stage results of an inverse-docking study which seeks to identify potential direct targets of PRIMA-1. PRIMA-1 is well known for its ability to restore mutant p53's tumor suppressor function, leading to apoptosis in several types of cancer cells. For this reason, we believe that potential direct targets of PRIMA-1 identified in silico should be experimentally screened for their ability to inhibitcancer cell growth. The highest-ranked human protein of our PRIMA-1 docking results is oxidosqualene cyclase (OSC), which is part of the cholesterol synthetic pathway. The results of two followup experiments which treat OSC as a possible anti-cancer target are promising. We show that both PRIMA-1 and Ro 48-8071, a known potent OSC inhibitor, significantly reduce theviability of BT-474 breast cancer cells relative to normal mammary cells. In addition, like PRIMA-1, we find that Ro 48-8071 results in increased binding of mutant p53 to DNA in BT- 474cells (which highly express p53). For the first time, Ro 48-8071 is shown as a potent agent in killing human breast cancer cells. The potential of OSC as a new target for developing anticancer therapies is worth further investigation.

Keywords: inverse docking, in silico screening, protein-ligand interactions, molecular docking

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1570 Virtual Screening and in Silico Toxicity Property Prediction of Compounds against Mycobacterium tuberculosis Lipoate Protein Ligase B (LipB)

Authors: Junie B. Billones, Maria Constancia O. Carrillo, Voltaire G. Organo, Stephani Joy Y. Macalino, Inno A. Emnacen, Jamie Bernadette A. Sy

Abstract:

The drug discovery and development process is generally known to be a very lengthy and labor-intensive process. Therefore, in order to be able to deliver prompt and effective responses to cure certain diseases, there is an urgent need to reduce the time and resources needed to design, develop, and optimize potential drugs. Computer-aided drug design (CADD) is able to alleviate this issue by applying computational power in order to streamline the whole drug discovery process, starting from target identification to lead optimization. This drug design approach can be predominantly applied to diseases that cause major public health concerns, such as tuberculosis. Hitherto, there has been no concrete cure for this disease, especially with the continuing emergence of drug resistant strains. In this study, CADD is employed for tuberculosis by first identifying a key enzyme in the mycobacterium’s metabolic pathway that would make a good drug target. One such potential target is the lipoate protein ligase B enzyme (LipB), which is a key enzyme in the M. tuberculosis metabolic pathway involved in the biosynthesis of the lipoic acid cofactor. Its expression is considerably up-regulated in patients with multi-drug resistant tuberculosis (MDR-TB) and it has no known back-up mechanism that can take over its function when inhibited, making it an extremely attractive target. Using cutting-edge computational methods, compounds from AnalytiCon Discovery Natural Derivatives database were screened and docked against the LipB enzyme in order to rank them based on their binding affinities. Compounds which have better binding affinities than LipB’s known inhibitor, decanoic acid, were subjected to in silico toxicity evaluation using the ADMET and TOPKAT protocols. Out of the 31,692 compounds in the database, 112 of these showed better binding energies than decanoic acid. Furthermore, 12 out of the 112 compounds showed highly promising ADMET and TOPKAT properties. Future studies involving in vitro or in vivo bioassays may be done to further confirm the therapeutic efficacy of these 12 compounds, which eventually may then lead to a novel class of anti-tuberculosis drugs.

Keywords: pharmacophore, molecular docking, lipoate protein ligase B (LipB), ADMET, TOPKAT

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1569 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

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1568 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

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1567 Clara Cell Secretory Protein 16 Serum Level Decreases in Patients with Non-Smoking-Related Chronic Obstructive Pulmonary Diseases (COPD)

Authors: Lian Wu, Mervyn Merrilees

Abstract:

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

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1566 Estimation of Genetic Diversity in Sorghum Accessions Using Agro-Mophological and Nutritional Traits

Authors: Maletsema Alina Mofokeng, Nemera Shargie

Abstract:

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

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1565 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

Abstract:

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

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1564 In vitro Anti-Gonococcal, Anti-Inflammatory and HIV-1 Reverse Transcriptase Activities of the Herbal Mixture

Authors: T. E. Tshikalange, B. C. Mophuting

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Traditional medicine often consists of complex ingredients prepared from a mixture of plant species. These herbal mixtures are used in the treatment of various ailments such as sexually transmitted diseases including HIV. The present study was carried out to determine the biological activities of the herbal mixture used traditionally in the treatment of sexually transmitted diseases. This herbal mixture consists of four plant species from families Asteraceae, Bignoniaceae, Fabaceae, and Myrtaceae. Five crude extracts (hexane, dichloromethane, methanol, water and boiled) of the herbal mixture were investigated for anti-gonococcal, anti-inflammatory, and reverse transcriptase activities. The anti-inflammatory activity of the plant extracts was determined by measuring the extract inhibitory effect on the pro-inflammatory enzyme lipoxygenase. The extracts were also tested for anti-HIV activity against recombinant HIV-1 enzyme using non-radioactive HIV-RT colorimetric assay. The boiled extract exhibited good anti-inflammatory activity with an IC₅₀ of 87 µg/ml compared to that of the positive control quercetin (IC₅₀= 92 µg/ml). All the other extracts showed little or no activity. Hexane extract was the only extract that showed reverse transcriptase extract inhibitory effect with an IC₅₀ of 74 µg/ml. Anti-gonococcal and cytotoxicity investigations are underway. The preliminary results support the use of herbal mixture by traditional healers.

Keywords: sexually transmitted diseases, lipoxygenase, anti-inflammatory, herbal mixture

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1563 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

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1562 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

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1561 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

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1560 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

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1559 Molecular Profiles of Microbial Etiologic Agents Forming Biofilm in Urinary Tract Infections of Pregnant Women by RTPCR Assay

Authors: B. Nageshwar Rao

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Urinary tract infection (UTI) represents the most commonly acquired bacterial infection worldwide, with substantial morbidity, mortality, and economic burden. The objective of the study is to characterize the microbial profiles of uropathogenic in the obstetric population by RTPCR. Study design: An observational cross-sectional study was performed at a single tertiary health care hospital among 50 pregnant women with UTIs, including asymptomatic and symptomatic patients attending the outpatient department and inpatient department of Obstetrics and Gynaecology.Methods: Serotyping and genes detection of various uropathogens were studied using RTPCR. Pulse filed gel electrophoresis methods were used to determine the various genetic profiles. Results: The present study shows that CsgD protein, involved in biofilm formation in Escherichia coli, VIM1, IMP1 genes for Klebsiella were identified by using the RTPCR method. Our results showed that the prevalence of VIM1 and IMP1 genes and CsgD protein in E.coli showed a significant relationship between strong biofilm formation, and this may be due to the prevalence of specific genes. Finally, the genetic identification of RTPCR results for both bacteria was correlated with each other and concluded that the above uropathogens were common isolates in producing Biofilm in the pregnant woman suffering from urinary tract infection in our hospital observational study.

Keywords: biofilms, Klebsiella, E.coli, urinary tract infection

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1558 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

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1557 In silico Designing and Insight into Antimalarial Potential of Chalcone-Quinolinylpyrazole Hybrids by Preclinical Study in Mice

Authors: Deepika Saini, Sandeep Jain, Ajay Kumar

Abstract:

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

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1556 Nutritional Value and Forage Quality Indicators in Some Rangeland’s Species at Different Vegetation Forms

Authors: Reza Dehghani Bidgoli

Abstract:

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

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1555 Proteomics Application in Disease Diagnosis and Reproduction İmprovement in Cow

Authors: Abdollah Sobhani, Hossein Vaseghi-Dodaran

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Proteomics is defined as the study of the component of a cell, tissue and biological fluid. This technique has the potential to identify protein biomarkers of a disease states. In this study which was performed on bovine ovarian follicular cysts (BOFC), eight proteins are over expressed in BOFC that these proteins could be useful biomarkers for BOFC. The difference between serum proteome pattern cows affected by postpartum endometritis with healthy cows revealed that concentrations orosomucoid was decreased in endometritis. The comparison proteome of brucella abortus between laboratory adapted strains and clinical isolates could be useful to better understand this disease and vaccine development. Proteomics experiments identified new proteins and pathways that may be important in future hypothesis-driven studies of glucocorticoid-induced immunosuppression. Understanding the molecular mechanisms of effective parameters on male fertility is essential for obtaining high reproductive efficiency by decreasing cost and time. The investigations on proteome of high fertility spermatozoa indicated that expression of some proteins such as casein kinase 2 (CKII) prime poly peptide and tyrosine kinase in high fertility spermatozoa was higher compared to low fertility spermatozoa. Also, some evidence has indicated that variation in protein types and amounts in seminal fluid regulates fertility indexes in dairy bull. In conclusion, proteomics is a useful technique for discovering drugs, vaccine development, and diagnosis disease by biomarkers and improvement of reproduction efficiency.

Keywords: proteomics, reproduction, biomarker, immunity

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1554 The Proximate Composition and Phytochemical Screening of Momordica Balsamina (Balsam Apple) Fruit and Leaves

Authors: Viruska Jaichand, John Jason Mellem, Viresh Mohanlall

Abstract:

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

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1553 Identification of Potential Small Molecule Regulators of PERK Kinase

Authors: Ireneusz Majsterek, Dariusz Pytel, J. Alan Diehl

Abstract:

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

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1552 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

Abstract:

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

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1551 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

Abstract:

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

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1550 YHV-Responsive Gene Expression under the Influence of PmRelish Regulation

Authors: Suwattana Visetnan, Premruethai Supungul, Sureerat Tang, Ikuo Hirono, Anchalee Tassanakajon, Vichien Rimphanitchayakit

Abstract:

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

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1549 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

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1548 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

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1547 Improving the Utilization of Telfairia occidentalis Leaf Meal with Cellulase-Glucanase-Xylanase Combination and Selected Probiotic in Broiler Diets

Authors: Ayodeji Fasuyi

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Telfairia occidentalis is a leafy vegetable commonly grown in the tropics for nutritional benefits. The use of enzymes and probiotics is becoming prominent due to the ban on antibiotics as growth promoters in many parts of the world. It is conceived that with enzymes and probiotics additives, fibrous leafy vegetables can be incorporated into poultry feeds as protein source. However, certain antinutrients were also found in the leaves of Telfairia occidentalis. Four broiler starter and finisher diets were formulated for the two phases of the broiler experiments. A mixture of fiber degrading enzymes, Roxazyme G2 (combination of cellulase, glucanase and xylanase) and probiotics (Turbotox), a growth promoter, were used in broiler diets at 1:1. The Roxazyme G2/Turbotox mixtures were used in diets containing four varying levels of Telfairia occidentalis leaf meal (TOLM) at 0, 10, 20 and 30%. Diets 1 were standard broiler diets without TOLM and Roxazyme G2 and Turbotox additives. Diets 2, 3 and 4 had enzymes and probiotics additives. Certain mineral elements such as Ca, P, K, Na, Mg, Fe, Mn, Cu and Zn were found in notable quantities viz. 2.6 g/100 g, 1.2 g/100 g, 6.2 g/100 g, 5.1 g/100 g, 4.7 g/100 g, 5875 ppm, 182 ppm, 136 ppm and 1036 ppm, respectively. Phytin, phytin-P, oxalate, tannin and HCN were also found in ample quantities viz. 189.2 mg/100 g, 120.1 mg/100 g, 80.7 mg/100 g, 43.1 mg/100 g and 61.2 mg/100 g, respectively. The average weight gain was highest at 46.3 g/bird/day for birds on 10% TOLM diet but similar (P > 0.05) to 46.2 g/bird/day for birds on 20% TOLM. The feed conversion ratio (FCR) of 2.27 was the lowest and optimum for birds on 10% TOLM although similar (P > 0.05) to 2.29 obtained for birds on 20% TOLM. FCR of 2.61 was the highest at 2.61 for birds on 30% TOLM diet. The lowest FCR of 2.27 was obtained for birds on 10% TOLM diet although similar (P > 0.05) to 2.29 for birds on 20% TOLM diet. Most carcass characteristics and organ weights were similar (P > 0.05) for the experimental birds on the different diets except for kidney, gizzard and intestinal length. The values for kidney, gizzard and intestinal length were significantly higher (P < 0.05) for birds on the TOLM diets. The nitrogen retention had the highest value of 72.37 ± 0.10% for birds on 10% TOLM diet although similar (P > 0.05) to 71.54 ± 1.89 obtained for birds on the control diet without TOLM and enzymes/probiotics mixture. There was evidence of a better utilization of TOLM as a plant protein source. The carcass characteristics and organ weights all showed evidence of uniform tissue buildup and muscles development particularly in diets containing 10% of TOLM level. There was also better nitrogen utilization in birds on the 10% TOLM diet. Considering the cheap cost of TOLM, it is envisaged that its introduction into poultry feeds as a plant protein source will ultimately reduce the cost of poultry feeds.

Keywords: Telfairia occidentalis leaf meal, enzymes, probiotics, additives

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1546 Medical Nutritional Therapy in Human Immunodeficiency Virus Infection with Tuberculosis and Severe Malnutrition: A Case Report

Authors: Lista Andriyati, Nurpudji A Taslim

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The human immunodeficiency virus (HIV) patients have potential nutritional and metabolic problems. HIV is a virus that attacks cells T helper and impairs the function of immune cells. Infected individuals gradually become immunodeficient, results in increased susceptibility to a wide range of infections such as tuberculosis (TB). Malnutrition has destructive effects on the immune system and host defense mechanisms. Effective and proper nutritional therapies are important to improve medical outcomes and quality of life, which is associated with functional improvement. A case of 38-years old man admitted to hospital with loss of consciousness and was diagnosed HIV infection and relapse lung TB with severe malnutrition, fever, oral candidiasis, anemia (6.3 g/dL), severe hypoalbuminemia (1.9 g/dL), severe hypokalemia (2.2 mmol/L), immune depletion (1085 /µL) and elevated liver enzyme (ALT 1198/AST 375 U/L). Nutritional intervention by giving 2300 kcal of energy, protein 2 g/IBW/day, carbohydrate 350 g, fat 104 g through enteral and parenteral nutrition. Supplementations administered are zinc, vitamin A, vitamin B1, vitamin B6, vitamin B12, vitamin C, vitamin D, and snakehead fish extract high content of protein albumin (Pujimin®). After 46 days, there are clinical and metabolic improvement in Hb (6.3 to 11.2 g/dL), potassium (2.2 to 3.4 mmol/L), albumin (1.9 to 2.3 g/dL), ALT 1198 to 47/AST 375 to 68 U/L) and improved awareness. In conclusion, nutritional therapy in HIV infection with adequate macronutrients and micronutrients fulfillment and immunonutrition is very important to avoid cachexia and to improve nutritional status and immune disfunction.

Keywords: HIV, hypoalbuminemia, malnutrition, tuberculosis

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1545 Deep Learning for Qualitative and Quantitative Grain Quality Analysis Using Hyperspectral Imaging

Authors: Ole-Christian Galbo Engstrøm, Erik Schou Dreier, Birthe Møller Jespersen, Kim Steenstrup Pedersen

Abstract:

Grain quality analysis is a multi-parameterized problem that includes a variety of qualitative and quantitative parameters such as grain type classification, damage type classification, and nutrient regression. Currently, these parameters require human inspection, a multitude of instruments employing a variety of sensor technologies, and predictive model types or destructive and slow chemical analysis. This paper investigates the feasibility of applying near-infrared hyperspectral imaging (NIR-HSI) to grain quality analysis. For this study two datasets of NIR hyperspectral images in the wavelength range of 900 nm - 1700 nm have been used. Both datasets contain images of sparsely and densely packed grain kernels. The first dataset contains ~87,000 image crops of bulk wheat samples from 63 harvests where protein value has been determined by the FOSS Infratec NOVA which is the golden industry standard for protein content estimation in bulk samples of cereal grain. The second dataset consists of ~28,000 image crops of bulk grain kernels from seven different wheat varieties and a single rye variety. In the first dataset, protein regression analysis is the problem to solve while variety classification analysis is the problem to solve in the second dataset. Deep convolutional neural networks (CNNs) have the potential to utilize spatio-spectral correlations within a hyperspectral image to simultaneously estimate the qualitative and quantitative parameters. CNNs can autonomously derive meaningful representations of the input data reducing the need for advanced preprocessing techniques required for classical chemometric model types such as artificial neural networks (ANNs) and partial least-squares regression (PLS-R). A comparison between different CNN architectures utilizing 2D and 3D convolution is conducted. These results are compared to the performance of ANNs and PLS-R. Additionally, a variety of preprocessing techniques from image analysis and chemometrics are tested. These include centering, scaling, standard normal variate (SNV), Savitzky-Golay (SG) filtering, and detrending. The results indicate that the combination of NIR-HSI and CNNs has the potential to be the foundation for an automatic system unifying qualitative and quantitative grain quality analysis within a single sensor technology and predictive model type.

Keywords: deep learning, grain analysis, hyperspectral imaging, preprocessing techniques

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