Search results for: whey protein isolated
Commenced in January 2007
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Search results for: whey protein isolated

2333 Efficacy of Mixed Actinomycetes against Fusarium Wilt Caused by Fusarium oxysporum f.sp. cubense

Authors: Jesryl B. Paulite, Irene Alcantara-Papa, Teofila O. Zulaybar, Jocelyn T. Zarate, Virgie Ugay

Abstract:

Banana is one of the major fruits in the Philippines in terms of volume of production and export earnings. The Philippines export of fresh Cavendish banana ranked No.1 with 22% share. One major threat to the industry is Fusarium wilt caused by Fusarium oxysporum f. sp. cubense. It tops as a major concern today affecting the Philippine banana industry since 2002 up to the present in Mindanao. Because of environmental and health issues concerning the use of chemical pesticides in the control of diseases, utilization of microorganisms has been significant in recent years as a promising alternative. This study aims to evaluate the potential of actinomycetes to control Fusarium wilt in Cavendish banana. The in-vitro experiments was carried out in Complete Randomized Design (CRD) while field experiment was laid out in a Randomized Complete Block Design (RCBD) with three treatments and three replications. Actinomycetes were isolated from mangrove soils in areas in Quezon and Bataan, Philippines. A total of 199 actinomycetes were isolated and 82 actinomycetes showed activity against the local Fusarium oxysporum (Foc) by agar plug assay. The test for antagonisms (AQ6, AQ30, and AQ121) of three best isolates Foc to were selected inhibiting Foc by 21.0mm, 22.0mm and 20.5mm, respectively. The same actinomycetes inhibited well Foc Tropical Race 4 showing 24.6 mm, 20.2mm and 19.0 mm zones of inhibition by agar plug assay, respectively. Combinations of the three isolates yielded an inhibition of 13.5 mm by cup cylinder assay. These findings led to the formulation of the mixed actinomycetes as biocontrol agents against Foc. A field experiment to evaluate the formulated mixed actinomycetes against Foc in a Foc infested field in Kinamayan, Sto Tomas, Davao Del Norte, Philippines. was conducted. Results showed that preventive method of application of the mixed actinomycetes against Foc showed promising results. A 56.66% mortality was observed in control set-up (no biocontrol agent added) compared to 33.33% mortality in preventive method. Further validation of the effectiveness of the mixed actinomycetes as biocontrol agent is presently being conducted in Asuncion, Davao Del Norte, Philippines.

Keywords: actinomycetes, biocontrol agents, cavendish banana, Fusarium oxysporum f. sp. cubense

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2332 Utilizing Bario Rice, a Natural Red-Pigmented Rice from Sarawak, Malaysia, in the Development of Gluten-Free Bread

Authors: Macdalyna Esther Ronie, Hasmadi Mamat, Ahmad Hazim Abdul Aziz, Mohamad Khairi Zainol

Abstract:

Current trends in gluten-free food development are increasingly leaning towards the utilization of pigmented rice flour, with a particular focus on Bario Merah Sederhana (BMS), a red-pigmented rice native to Sarawak, Malaysia. This study delves into the evaluation of the nutritional, textural, and sensory attributes of gluten-free rice bread produced from a blend of BMS rice flour and potato starch. The resulting samples are denoted as F1 (100% BMS rice flour), F2 (90% BMS rice flour and 10% potato starch), F3 (80% BMS rice flour and 20% potato starch), and F4 (70% BMS rice flour and 30% potato starch). Comparatively, these gluten-free rice bread formulations exhibit higher levels of ash and crude fiber, along with lower carbohydrate content when juxtaposed with conventional wheat bread. Notably, the crude protein content of the rice bread diminishes significantly (p<0.05) as the proportion of rice flour decreases, primarily due to the higher protein content found in wheat flour. The crumb of the rice bread appears darker owing to the red pigment in the rice flour, while the crust is lighter than that of the control sample, possibly attributable to a reduced Maillard reaction. Among the various rice bread formulations, F4 stands out with the least dough and bread hardness, accompanied by the highest levels of stickiness and springiness in both dough and bread, respectively. In sensory evaluations, wheat bread garners the highest rating (p<0.05). However, within the realm of rice breads, F4 emerges as a viable and acceptable formulation, as indicated by its commendable scores in color (7.03), flavor (5.73), texture (6.03), and overall acceptability (6.18). These findings underscore the potential of BMS in the creation of gluten-free rice breads, with the formulation consisting of 70% rice flour and 30% potato starch emerging as a well-received and suitable option.

Keywords: gluten-free bread, bario rice, proximate composition, sensory evaluation

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2331 Actinomycetes from Protected Forest Ecosystems of Assam, India: Diversity and Antagonistic Activity

Authors: Priyanka Sharma, Ranjita Das, Mohan C. Kalita, Debajit Thakur

Abstract:

Background: Actinomycetes are the richest source of novel bioactive secondary metabolites such as antibiotics, enzymes and other therapeutically useful metabolites with diverse biological activities. The present study aims at the antimicrobial potential and genetic diversity of culturable Actinomycetes isolated from protected forest ecosystems of Assam which includes Kaziranga National Park (26°30˝-26°45˝N and 93°08˝-93°36˝E), Pobitora Wildlife Sanctuary (26º12˝-26º16˝N and 91º58˝-92º05˝E) and Gibbon Wildlife Sanctuary (26˚40˝-26˚45˝N and 94˚20˝-94˚25˝E) which are located in the North-eastern part of India. Northeast India is a part of the Indo-Burma mega biodiversity hotspot and most of the protected forests of this region are still unexplored for the isolation of effective antibiotic-producing Actinomycetes. Thus, there is tremendous possibility that these virgin forests could be a potential storehouse of novel microorganisms, particularly Actinomycetes, exhibiting diverse biological properties. Methodology: Soil samples were collected from different ecological niches of the protected forest ecosystems of Assam and Actinomycetes were isolated by serial dilution spread plate technique using five selective isolation media. Preliminary screening of Actinomycetes for an antimicrobial activity was done by spot inoculation method and the secondary screening by disc diffusion method against several test pathogens, including multidrug resistant Staphylococcus aureus (MRSA). The strains were further screened for the presence of antibiotic synthetic genes such as type I polyketide synthases (PKS-I), type II polyketide synthases (PKS-II) and non-ribosomal peptide synthetases (NRPS) genes. Genetic diversity of the Actinomycetes producing antimicrobial metabolites was analyzed through 16S rDNA-RFLP using Hinf1 restriction endonuclease. Results: Based on the phenotypic characterization, a total of 172 morphologically distinct Actinomycetes were isolated and screened for antimicrobial activity by spot inoculation method on agar medium. Among the strains tested, 102 (59.3%) strains showed activity against Gram-positive bacteria, 98 (56.97%) against Gram-negative bacteria, 92 (53.48%) against Candida albicans MTCC 227 and 130 (75.58%) strains showed activity against at least one of the test pathogens. Twelve Actinomycetes exhibited broad spectrum antimicrobial activity in the secondary screening. The taxonomic identification of these twelve strains by 16S rDNA sequencing revealed that Streptomyces was found to be the predominant genus. The PKS-I, PKS-II and NRPS genes detection indicated diverse bioactive products of these twelve Actinomycetes. Genetic diversity by 16S rDNA-RFLP indicated that Streptomyces was the dominant genus amongst the antimicrobial metabolite producing Actinomycetes. Conclusion: These findings imply that Actinomycetes from the protected forest ecosystems of Assam, India, are a potential source of bioactive secondary metabolites. These areas are as yet poorly studied and represent diverse and largely unscreened ecosystem for the isolation of potent Actinomycetes producing antimicrobial secondary metabolites. Detailed characterization of the bioactive Actinomycetes as well as purification and structure elucidation of the bioactive compounds from the potent Actinomycetes is the subject of ongoing investigation. Thus, to exploit Actinomycetes from such unexplored forest ecosystems is a way to develop bioactive products.

Keywords: Actinomycetes, antimicrobial activity, forest ecosystems, RFLP

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2330 Enhanced Anti-Obesity Effect of Soybean by Fermentation with Lactobacillus plantarum P1201 in 3T3-L1 Adipocyte

Authors: Chengliang Xie, Jinhyun Ryu, Hyun Joon Kim, Gyeong Jae Cho, Wan Sung Choi, Sang Soo Kang, Kye Man Cho, Dong Hoon Lee

Abstract:

Obesity has become a global health problem and a source of major metabolic diseases like type-2 diabetes, hypertension, heart disease, nonalcoholic fatty liver and cancer. Synthetic anti-obesity drugs are effective but very costly and with undesirable side effects, so natural products such as soybean are needed as an alternative for obesity treatment. Lactobacillus Plantarum P1201is a probiotic bacterial strain reported to produce conjugated linoleic acid (CLA) and increase the ratio of aglycone-isoflavone of soybean, both of which have anti-obesity effect. In this study, the anti-obesity effect of the fermented soybean extract with P1201 (FSE) will be evaluated compared with that of the soybean extract (SE) by 3T3-L1 cells as an in vitro model of adipogenesis. 3T3-L1 cells were treated with SE and FSE during the nine days of the differentiation, lipid accumulation was evaluated by oil-red staining and triglyceride content and the mRNA expression level of adipogenic or lipogenic genes were analyzed by RT-PCR and qPCR. The results showed that formation of lipid droplets in differentiated 3T3-L1 cells was inhibited and triglyceride content was reduced by 23.1% after treated with 1000 μg/mL of FSE compared with control. For SE-treated groups, no delipidating effect was observed. The effect of FSE on adipogenesis inhibition can be attributed to the down-regulation of mRNA expressionof CCAAT/enhancer binding protein (C/EBP-α), lipoprotein lipase (LPL), adiponectin, adipocyte fatty acid-binding protein (aP2), fatty acid synthesis (FAS) and CoA carboxylase (ACC). Our results demonstrated that the anti-obesity effect of soybean can be improved by fermentation with P1201, and P1201can be used as a potential probiotic bacterial strain to produce natural anti-obesity food.

Keywords: fermentation, Lactobacillus plantarum P1201, obesity, soybean

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2329 Fatty Acid Binding Protein 3 Gene Polymorphisms and Their Associations with Growth Traits and Blood Parameters in Two Iranian Sheep Breeds

Authors: Sahar Javadi-Novashnagh, Mohammad Moradi-Shahrbabak, Mostafa Sadeghi, Katarzyna Ropka-Molik, Hossein Moradi-Shahrbabak, Maria Consuelo Mura

Abstract:

The objective of this study was to investigate two single nucleotide polymorphisms located in exon 2 (g.939A > G) and intron 3 (g.4349A > G) of fatty acid binding protein 3 (FABP3) gene in two Iranian sheep breeds, Lori-Bakhtiari and Zel, using polymerase chain reaction -restriction fragment length polymorphism (PCR-RFLP) approach. The association of the polymorphisms with growth traits and blood parameters was also examined. Results revealed a g.939A > G SNP (single nucleotide polymorphism) in the exon 2 exhibiting three genotypes: AA, AG, and GG. Statistical analysis indicated that this polymorphism significantly influenced blood triglyceride (P < 0.05) and cholesterol (P < 0.08) levels as well as weaning weight (P < 0.05). Animals with AG genotype had the highest blood triglyceride level and weaning weight while the highest amount of blood cholesterol was observed in animals with GG genotype. On the other hand, no significant effect was observed on birth and fat-tail weight traits. The intron 3 (g.4349A > G) was monomorphic across the studied samples. Lori-Bakhtiari breed showed significantly higher blood triglyceride and cholesterol levels, as also birth and weaning weight compared to Zel breed (P < 0.01). Considering that the literature is bereft of any report on the association study between FABP3 SNPs and sheep growth traits and blood parameters, our findings suggest that the investigated polymorphism might be one of the main genetic factors affecting growth and physiological traits in sheep.

Keywords: FABP3 gene, fatness, weaning weight, blood triglyceride, cholesterol, Zel, Lori-Bakhtiari

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2328 Phytoremediation of Heavy Metals by Phragmites Australis at Oeud Meboudja Annaba Algeria

Authors: Kleche Myriam, Ziane Nadia, Berrebbah Houria, Djebar Mohammed Reda

Abstract:

The Phytoremediation has now become a necessity. Thus, in our work, we are interested in the biological wastewater treatment of Oued Meboudja. The physicochemical analysis of water after treatment showed a significant reduction of suspended matter, COD and BOD5 and rate of metals in roots for example iron and zinc. We also highlighted some significant changes in biometric and physiological parameters such as increasing the number of roots and increased respiratory metabolism through the oxygen consumption in isolated roots of Phragmites australis, placed in a polluted environment.

Keywords: phragmites australis, roots, phytoremediation, iron, zinc

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2327 Evaluation of Some Serum Proteins as Markers for Myeloma Bone Disease

Authors: V. T. Gerov, D. I. Gerova, I. D. Micheva, N. F. Nazifova-Tasinova, M. N. Nikolova, M. G. Pasheva, B. T. Galunska

Abstract:

Multiple myeloma (MM) is the most frequent plasma cell (PC) dyscrasia that involves the skeleton. Myeloma bone disease (MBD) is characterized by osteolytic bone lesions as a result of increased osteoclasts activity not followed by reactive bone formation due to osteoblasts suppression. Skeletal complications cause significant adverse effects on quality of life and lead to increased morbidity and mortality. Last decade studies revealed the implication of different proteins in osteoclast activation and osteoblast inhibition. The aim of the present study was to determine serum levels of periostin, sRANKL and osteopontin and to evaluate their role as bone markers in MBD. Materials and methods. Thirty-two newly diagnosed MM patients (mean age: 62.2 ± 10.7 years) and 33 healthy controls (mean age: 58.9 ± 7.5 years) were enrolled in the study. According to IMWG criteria 28 patients were with symptomatic MM and 4 with monoclonal gammopathy of undetermined significance (MGUS). In respect to their bone involvement all symptomatic patients were divided into two groups (G): 9 patients with 0-3 osteolytic lesions (G1) and 19 patients with >3 osteolytic lesions and/or pathologic fractures (G2). Blood samples were drawn for routine laboratory analysis and for measurement of periostin, sRANKL and osteopontin serum levels by ELISA kits (Shanghai Sunred Biological Technology, China). Descriptive analysis, Mann-Whitney test for assessment the differences between groups and non-parametric correlation analysis were performed using GraphPad Prism v8.01. Results. The median serum levels of periostin, sRANKL and osteopontin of ММ patients were significantly higher compared to controls (554.7pg/ml (IQR=424.0-720.6) vs 396.9pg/ml (IQR=308.6-471.9), p=0.0001; 8.9pg/ml (IQR=7.1-10.5) vs 5.6pg/ml (IQR=5.1-6.4, p<0.0001 and 514.0ng/ml (IQR=469.3-754.0) vs 387.0ng/ml (IQR=335.9-441.9), p<0.0001, respectively). for assessment of differences between groups and non-parametric correlation analysis were performed using GraphPad Prism v8.01. Statistical significance was found for all tested bone markers between symptomatic MM patients and controls: G1 vs controls (p<0.03), G2 vs controls (p<0.0001) for periostin; G1 vs controls (p<0.0001), G2 vs controls (p<0.0001) for sRANKL; G1 vs controls (p=0.002), G2 vs controls (p<0.0001) for osteopontin, as well between symptomatic MM patients and MGUS patients: G1 vs MGUS (p<0.003), G2 vs MGUS (p=0.003) for periostin; G1 vs MGUS (p<0.05), G2 vs MGUS (p<0.001) for sRANKL; G1 vs MGUS (p=0.011), G2 vs MGUS (p=0.0001) for osteopontin. No differences were detected between MGUS and controls and between patients in G1 and G2 groups. Spearman correlation analysis revealed moderate positive correlation between periostin and beta-2-microglobulin (r=0.416, p=0.018), percentage bone marrow myeloma PC (r=0.432, p=0.014), and serum total protein (r=0.427, p=0.015). Osteopontin levels were also positively related to beta-2-microglobulin (r=0.540, p=0.0014), percentage bone marrow myeloma PC (r=0.423, p=0.016), and serum total protein (r=0.413, p=0.019). Serum sRANKL was only related to beta-2-microglobulin levels (r=0.398, p=0.024). Conclusion: In the present study, serum levels of periostin, sRANKL and osteopontin in newly diagnosed MM patients were evaluated. They gradually increased from MGUS to more advanced stages of MM reflecting the severity of bone destruction. These results support the idea that some new protein markers could be used in monitoring the MBD as a most severe complication of MM.

Keywords: myeloma bone disease, periostin, sRANKL, osteopontin

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2326 Chemical Composition of the Essential Oil of Citrus aurantium Isolated by Solvent Free Microwave Assisted Extraction and Hydrodistillation Extraction

Authors: Masume Rezaie, Mohammad H. Farjam

Abstract:

Chemical composition of Citrus aurantium was studied by solvent free microwave extraction (SFME) and hydrodistillation (HD) methods. Limonene (76.06% SFME and 67.04% HD), Linalool (4.91% SFME and 10.08% HD) and Linalyl Acetate (8.52% SFME and 5.10% HD) were the major compounds that obtained by SFME and hydrodistillation, respectively.

Keywords: microwave-assisted, GC-MS, essential oils, hydrodistillation, citrus aurantium

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2325 Molecular Characterization and Arsenic Mobilization Properties of a Novel Strain IIIJ3-1 Isolated from Arsenic Contaminated Aquifers of Brahmaputra River Basin, India

Authors: Soma Ghosh, Balaram Mohapatra, Pinaki Sar, Abhijeet Mukherjee

Abstract:

Microbial role in arsenic (As) mobilization in the groundwater aquifers of Brahmaputra river basin (BRB) in India, severely threatened by high concentrations of As, remains largely unknown. The present study, therefore, is a molecular and ecophysiological characterization of an indigenous bacterium strain IIIJ3-1 isolated from As contaminated groundwater of BRB and application of this strain in several microcosm set ups differing in their organic carbon (OC) source and terminal electron acceptors (TEA), to understand its role in As dissolution under aerobic and anaerobic conditions. Strain IIIJ3-1 was found to be a new facultative anaerobic, gram-positive, endospore-forming strain capable of arsenite (As3+) oxidation and dissimilatory arsenate (As5+) reduction. The bacterium exhibited low genomic (G+C)% content (45 mol%). Although, its 16S rRNA gene sequence revealed a maximum similarity of 99% with Bacillus cereus ATCC 14579(T) but the DNA-DNA relatedness of their genomic DNAs was only 49.9%, which remains well below the value recommended to delimit different species. Abundance of fatty acids iC17:0, iC15:0 and menaquinone (MK) 7 though corroborates its taxonomic affiliation with B. cereus sensu-lato group, presence of hydroxy fatty acids (HFAs), C18:2, MK5 and MK6 marked its uniqueness. Besides being highly As resistant (MTC=10mM As3+, 350mM As5+), metabolically diverse, efficient aerobic As3+ oxidizer; it exhibited near complete dissimilatory reduction of As5+ (1 mM). Utilization of various carbon sources with As5+ as TEA revealed lactate to serve as the best electron donor. Aerobic biotransformation assay yielded a lower Km for As3+ oxidation than As5+ reduction. Arsenic homeostasis was found to be conferred by the presence of arr, arsB, aioB, and acr3(1) genes. Scanning electron microscopy (SEM) coupled with energy dispersive X-ray (EDX) analysis of this bacterium revealed reduction in cell size upon exposure to As and formation of As-rich electron opaque dots following growth with As3+. Incubation of this strain with sediment (sterilised) collected from BRB aquifers under varying OC, TEA and redox conditions revealed that the strain caused highest As mobilization from solid to aqueous phase under anaerobic condition with lactate and nitrate as electron donor and acceptor, respectively. Co-release of highest concentrations of oxalic acid, a well known bioweathering agent, considerable fold increase in viable cell counts and SEM-EDX and X-ray diffraction analysis of the sediment after incubation under this condition indicated that As release is consequent to microbial bioweathering of the minerals. Co-release of other elements statistically proves decoupled release of As with Fe and Zn. Principle component analysis also revealed prominent role of nitrate under aerobic and/or anaerobic condition in As release by strain IIIJ3-1. This study, therefore, is the first to isolate, characterize and reveal As mobilization property of a strain belonging to the Bacillus cereus sensu lato group isolated from highly As contaminated aquifers of Brahmaputra River Basin.

Keywords: anaerobic microcosm, arsenic rich electron opaque dots, Arsenic release, Bacillus strain IIIJ3-1

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2324 Contribution of mTOR to Oxidative/Nitrosative Stress via NADPH Oxidase System Activation in Zymosan-Induced Systemic Inflammation in Rats

Authors: Seyhan Sahan-Firat, Meryem Temiz-Resitoglu, Demet Sinem Guden, Sefika Pinar Kucukkavruk, Bahar Tunctan, Ayse Nihal Sari, Zumrut Kocak

Abstract:

We hypothesized that mTOR inhibition may prevent the multiple organ failures following severe multiple tissue injury associated with increased NADPH oxidase system activity occur in zymosan-induced systemic inflammation. Therefore, we investigated the role of mTOR in oxidative/nitrosative stress associated with increase in NADPH oxidase activity in zymosan-induced systemic inflammation model in rats. Male Wistar rats received saline (4 ml/kg, i.p.) and zymosan (500 mg/kg, i.p.) at time 0. Saline, or zymosan-treated rats were given rapamycin (1 mg/kg, i.p.) 1 h after saline or zymosan injections. Rats were sacrified 4 h after zymosan challenge and kidney, heart, thoracic aorta, and superior mesenteric artery were collected. NADPH oxidase activity, p22phox, gp91phox, and p47phox protein expression and nitrotyrosine levels were measured in tissue samples. Zymosan administration caused an increase in NADPH oxidase activity, p22phox, gp91phox, and p47phox protein expression and nitrotyrosine levels in kidney, heart, thoracic aorta, and superior mesenteric artery. These changes caused by zymosan reversed by rapamycin, a selective mTOR inhibitor. Rapamycin alone had no effect on the parameters measured. Our results demonstrated that zymosan-induced oxidative/nitrosative stress presumably due to enhanced activity of NADPH oxidase, expression of p22phox, gp91phox, and p47phox and production of peroxynitrite were mediated by mTOR. [This work was financially supported by Research Foundation of Mersin University (2016-2-AP3-1900)].

Keywords: oxidative stress, mTOR, nitrosative stress, zymosan

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2323 Identification of Analogues to EGCG for the Inhibition of HPV E7: A Fundamental Insights through Structural Dynamics Study

Authors: Murali Aarthy, Sanjeev Kumar Singh

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High risk human papillomaviruses are highly associated with the carcinoma of the cervix and the other genital tumors. Cervical cancer develops through the multistep process in which increasingly severe premalignant dysplastic lesions called cervical intraepithelial neoplastic progress to invasive cancer. The oncoprotein E7 of human papillomavirus expressed in the lower epithelial layers drives the cells into S-phase creating an environment conducive for viral genome replication and cell proliferation. The replication of the virus occurs in the terminally differentiating epithelium and requires the activation of cellular DNA replication proteins. To date, no suitable drug molecule is available to treat HPV infection whereas identification of potential drug targets and development of novel anti-HPV chemotherapies with unique mode of actions are expected. Hence, our present study aimed to identify the potential inhibitors analogous to EGCG, a green tea molecule which is considered to be safe to use for mammalian systems. A 3D similarity search on the natural small molecule library from natural product database using EGCG identified 11 potential hits based on their similarity score. The structure based docking strategies were implemented in the potential hits and the key interacting residues of protein with compounds were identified through simulation studies and binding free energy calculations. The conformational changes between the apoprotein and the complex were analyzed with the simulation and the results demonstrated that the dynamical and structural effects observed in the protein were induced by the compounds and indicated the dominance to the oncoprotein. Overall, our study provides the basis for the structural insights of the identified potential hits and EGCG and hence, the analogous compounds identified can be potent inhibitors against the HPV 16 E7 oncoprotein.

Keywords: EGCG, oncoprotein, molecular dynamics simulation, analogues

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2322 Whole Exome Sequencing Data Analysis of Rare Diseases: Non-Coding Variants and Copy Number Variations

Authors: S. Fahiminiya, J. Nadaf, F. Rauch, L. Jerome-Majewska, J. Majewski

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Background: Sequencing of protein coding regions of human genome (Whole Exome Sequencing; WES), has demonstrated a great success in the identification of causal mutations for several rare genetic disorders in human. Generally, most of WES studies have focused on rare variants in coding exons and splicing-sites where missense substitutions lead to the alternation of protein product. Although focusing on this category of variants has revealed the mystery behind many inherited genetic diseases in recent years, a subset of them remained still inconclusive. Here, we present the result of our WES studies where analyzing only rare variants in coding regions was not conclusive but further investigation revealed the involvement of non-coding variants and copy number variations (CNV) in etiology of the diseases. Methods: Whole exome sequencing was performed using our standard protocols at Genome Quebec Innovation Center, Montreal, Canada. All bioinformatics analyses were done using in-house WES pipeline. Results: To date, we successfully identified several disease causing mutations within gene coding regions (e.g. SCARF2: Van den Ende-Gupta syndrome and SNAP29: 22q11.2 deletion syndrome) by using WES. In addition, we showed that variants in non-coding regions and CNV have also important value and should not be ignored and/or filtered out along the way of bioinformatics analysis on WES data. For instance, in patients with osteogenesis imperfecta type V and in patients with glucocorticoid deficiency, we identified variants in 5'UTR, resulting in the production of longer or truncating non-functional proteins. Furthermore, CNVs were identified as the main cause of the diseases in patients with metaphyseal dysplasia with maxillary hypoplasia and brachydactyly and in patients with osteogenesis imperfecta type VII. Conclusions: Our study highlights the importance of considering non-coding variants and CNVs during interpretation of WES data, as they can be the only cause of disease under investigation.

Keywords: whole exome sequencing data, non-coding variants, copy number variations, rare diseases

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2321 DeepLig: A de-novo Computational Drug Design Approach to Generate Multi-Targeted Drugs

Authors: Anika Chebrolu

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Mono-targeted drugs can be of limited efficacy against complex diseases. Recently, multi-target drug design has been approached as a promising tool to fight against these challenging diseases. However, the scope of current computational approaches for multi-target drug design is limited. DeepLig presents a de-novo drug discovery platform that uses reinforcement learning to generate and optimize novel, potent, and multitargeted drug candidates against protein targets. DeepLig’s model consists of two networks in interplay: a generative network and a predictive network. The generative network, a Stack- Augmented Recurrent Neural Network, utilizes a stack memory unit to remember and recognize molecular patterns when generating novel ligands from scratch. The generative network passes each newly created ligand to the predictive network, which then uses multiple Graph Attention Networks simultaneously to forecast the average binding affinity of the generated ligand towards multiple target proteins. With each iteration, given feedback from the predictive network, the generative network learns to optimize itself to create molecules with a higher average binding affinity towards multiple proteins. DeepLig was evaluated based on its ability to generate multi-target ligands against two distinct proteins, multi-target ligands against three distinct proteins, and multi-target ligands against two distinct binding pockets on the same protein. With each test case, DeepLig was able to create a library of valid, synthetically accessible, and novel molecules with optimal and equipotent binding energies. We propose that DeepLig provides an effective approach to design multi-targeted drug therapies that can potentially show higher success rates during in-vitro trials.

Keywords: drug design, multitargeticity, de-novo, reinforcement learning

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2320 Nephroprotective Effect of Aqueous Extract of Plectranthus amboinicus (Roxb.) Leaves in Adriamycin Induced Acute Renal Failure in Wistar Rats: A Biochemical and Histopathological Assessment

Authors: Ampe Mohottige Sachinthi Sandaruwani Amarasiri, Anoja Priyadarshani Attanayake, Kamani Ayoma Perera Wijewardana Jayatilaka, Lakmini Kumari Boralugoda Mudduwa

Abstract:

The search for alternative pharmacological therapies based on natural extracts for renal failure has become an urgent need, due to paucity of effective pharmacotherapy. The current study was undertaken to evaluate the acute nephroprotective effect of aqueous leaf extract of Plectranthus amboinicus (Roxb.) (Family: Lamiaceae), a medicinal plant used in traditional Ayurvedic medicine for the management of renal diseases in Sri Lanka. The study was performed in adriamycin (ADR) induced nephrotoxic in Wistar rats. Wistar rats were randomly divided into four groups each with six rats. A single dose of ADR (20 mg/kg body wt., ip) was used for the induction of nephrotoxicity in all groups of rats except group one. The treatments were started 24 hours after induction of nephrotoxicity and continued for three days. Group one and two served as healthy and nephrotoxic control rats and were administered equivalent volumes of normal saline (0.9% NaCl) orally. Group three and four nephrotoxic rats were administered the lyophilized powder of the aqueous extract of P. amboinicus (400 mg/ kg body wt.; equivalent human therapeutic dose) and the standard drug, fosinopril sodium (0.09 mg/ kg body wt.) respectively. Urine and blood samples were collected from rats in each group at the end of the period of intervention for the estimation of selected renal parameters. H and E stained sections of the kidney tissues were examined for histopathological changes. Rats treated with the plant extract showed significant improvement in biochemical parameters and histopathological changes compared to ADR induced nephrotoxic group. The elevation of serum concentrations of creatinine and β2-microglobulin were decreased by 38%, and 66% in plant extract treated nephrotoxic rats respectively (p < 0.05). In addition, serum concentrations of total protein and albumin were significantly increased by 25% and 14% in rats treated with P. amboinicus respectively (p < 0.05). The results of β2 –microglobulin and serum total protein demonstrated a significant reduction in the elevated values in rats administered with the plant extract (400 mg/kg) compared to that of fosinopril (0.09 mg/kg). Urinary protein loss in 24hr urine samples was significantly decreased in rats treated with both fosinopril (86%) and P. ambonicus (56%) at the end of the intervention (p < 0.01). Accordingly, an attenuation of morphological destruction was observed in the H and E stained sections of the kidney with the treatments of plant extract and fosinopril. The results of the present study revealed that the aqueous leaf extract of P. amboinicus possesses significant nephroprotective activity at the equivalent therapeutic dose of 400 mg/ kg against adriamycin induced acute nephrotoxicity.

Keywords: biochemical assessment, histopathological assessment, nephroprotective activity, Plectranthus amboinicus

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2319 Isolation, Identification and Screening of Pectinase Producing Fungi Isolated from Apple (Malus Domestica)

Authors: Shameel Pervez, Saad Aziz Durrani, Ibatsam Khokhar

Abstract:

Pectinase is an enzyme that breaks down pectin, a compound responsible for structural integrity of the plant. Pectin is difficult to break down mechanically and the cost is very high, that is why many industries including food industries use pectinase enzyme produced by microbes for pectin breakdown. Apple (Malus domestica) is an important fruit in terms of market value. Every year, millions of apples are wasted due to post-harvest rot caused by fungi. Fungi are natural decomposers of our ecosystem and are infamous for post-harvest rot of apple fruit but at the same time they are prized for their high production of valuable extracellular enzymes such as pectinase. In this study, fungi belonging to different genus were isolated from rotten apples. Rotten samples of apple were picked from different markets of Lahore. After surface sterilization, the rotten parts were cut into small pieces and placed onto MEA media plates for three days. Afterwards, distinct colonies were picked and purified by sub-culturing. The isolates were identified to genus level through the study of basic colony morphology and microscopic features. The isolates were then subjected to screening for pectinase activity on MS media to compare pectinase production and were then subsequently tested for pathogenic activity through wound suspension method to evaluate the pathogenic activity of isolates in comparison with their pectinolytic activity. A total of twelve fungal strains were isolates from rotten apples. They were belonging to genus Penicillium, Alternaria, Paecilomyces and Rhizopus. Upon screening for pectinolytic activity, isolates Pen 1, Pen 4, and Rz showed high pectinolytic activity and were further subjected to DNA isolation and partial sequencing for species identification. The results of partial sequencing were combined with in-depth study of morphological features revealing Pen 1 as Penicillium janthinellum, Pen 4 as Penicillium griseofulvum, and Rz as Rhizopus microsporus. Pathogenic activity of all twelve isolates was evaluated. Penicillium spp. were highly pathogenic and destructive and same was the case with Paecilomyces sp. and Rhizopus sp. However, Alternaria spp. were found to be more consistent in their pathogenic activity, on all types of apples.

Keywords: apple, pectinase, fungal pathogens, penicillium, rhizopus

Procedia PDF Downloads 42
2318 Seed Yield and Quality of Late Planted Rabi Wheat Crop as Influenced by Basal and Foliar Application of Urea

Authors: Omvati Verma, Shyamashrre Roy

Abstract:

A field experiment was conducted with three basal nitrogen levels (90, 120 and 150 kg N/ha) and five foliar application of urea (absolute control, water spray, 3% urea spray at anthesis, 7 and 14 days after anthesis) at G.B. Pant University of Agriculture & Technology, Pantnagar, U.S. Nagar (Uttarakhand) during rabi season in a factorial randomized block design with three replications. Results revealed that nitrogen application of 150 kg/ha produced the highest seed yield, straw and biological yield and it was significantly superior to 90 kg N/ha and was at par with 120 kg N/ha. The number of tillers increased significantly with increase in nitrogen doses up to 150 kg N/ha. Spike length, number of grains per spike, grain weight per spike and thousand seed weight showed significantly higher values with 120 kg N/ha than 90 kg N/ha and were at par with that of 150 kg N/ha. Also, plant height showed similar trend. Leaf area index and chlorophyll content showed significant increase with an increase in nitrogen levels at different stages. In the case of foliar spray treatments, urea spray at anthesis showed highest value for yield and yield attributes. In case of spike length and thousand seed weight, it was similar with the urea spray at 7 and 14 days after anthesis, but for rest of the yield attributes, it was significantly higher than rest of the treatments. Among seed quality parameters protein and sedimentation value showed significant increase due to increase in nitrogen rates whereas, starch and hectolitre weight had a decreasing trend. Wet gluten content was not influenced by nitrogen levels. Foliar urea spray at anthesis resulted in highest value of protein and hectolitre weight whereas, urea spray at 7 days after anthesis showed highest value of sedimentation value and wet gluten content.

Keywords: foliar application, nitrogenous fertilizer, seed quality, yield

Procedia PDF Downloads 257
2317 Nutrition Intervention for Spinal Cord Injury in Critical Care

Authors: Dina Muharib

Abstract:

Specific metabolic challenges are present following spinal cord injury. The acute stage is characterized by a reduction in metabolic activity, as well as a negative nitrogen balance that cannot be corrected, even with aggressive nutritional support. Metabolic demands need to be accurately monitored to avoid overfeeding. Enteral feeding is the optimal route following SCI. When oral feeding is not possible, nasogastric, followed by nasojejunal, then by percutaneous endoscopic gastrostomy, if necessary, is suggested.

Keywords: SCI, energy, protein, nutrition assessment, eneral feeding, nitrogen balance

Procedia PDF Downloads 448
2316 Proliferative Effect of Some Calcium Channel Blockers on the Human Embryonic Kidney Cell Line

Authors: Lukman Ahmad Jamil, Heather M. Wallace

Abstract:

Introduction: Numerous epidemiological studies have shown a positive as well as negative association and no association in some cases between chronic use of calcium channel blockers and the increased risk of developing cancer. However, these associations were enmeshed with controversies in the absence of laboratory based studies to back up those claims. Aim: The aim of this study was to determine in mechanistic terms the association between the long-term administration of nifedipine and diltiazem and increased risk of developing cancer using the human embryonic kidney (HEK293) cell line. Methods: Cell counting using the Trypan blue dye exclusion and 3-4, 5-Dimethylthiazol-2-yl-2, 5-diphenyl-tetrazolium bromide (MTT) assays were used to investigate the effect of nifedipine and diltiazem on the growth pattern of HEK293 cells. Protein assay using modified Lowry method and analysis of intracellular polyamines concentration using Liquid Chromatography – Tandem Mass Spectrometry (LC-MS) were performed to ascertain the mechanism through which chronic use of nifedipine increases the risk of developing cancer. Results: Both nifedipine and diltiazem significantly increased the proliferation of HEK293 cells dose and time dependently. This proliferative effect after 24, 48 and 72-hour incubation period was observed at 0.78, 1.56 and 25 µM for nifedipine and 0.39, 1.56 and 25 µM for diltiazem, respectively. The increased proliferation of the cells was found to be statistically significantly (p<0.05). Furthermore, the increased proliferation of the cells induced by nifedipine was associated with the increase in the protein content and elevated intracellular polyamines concentration level. Conclusion: The chronic use of nifedipine is associated with increased proliferation of cells with concomitant elevation of polyamines concentration and elevated polyamine levels have been implicated in many malignant transformations and hence, these provide a possible explanation on the link between long term use of nifedipine and development of some human cancers. Further studies are needed to evaluate the cause of this association.

Keywords: cancer, nifedipine, polyamine, proliferation

Procedia PDF Downloads 180
2315 Virulence Factors and Drug Resistance of Enterococci Species Isolated from the Intensive Care Units of Assiut University Hospitals, Egypt

Authors: Nahla Elsherbiny, Ahmed Ahmed, Hamada Mohammed, Mohamed Ali

Abstract:

Background: The enterococci may be considered as opportunistic agents particularly in immunocompromised patients. It is one of the top three pathogens causing many healthcare associated infections (HAIs). Resistance to several commonly used antimicrobial agents is a remarkable characteristic of most species which may carry various genes contributing to virulence. Objectives: to determine the prevalence of enterococci species in different intensive care units (ICUs) causing health care-associated infections (HAIs), intestinal carriage and environmental contamination. Also, to study the antimicrobial susceptibility pattern of the isolates with special reference to vancomycin resistance. In addition to phenotypic and genotypic detection of gelatinase, cytolysin and biofilm formation among isolates. Patients and Methods: This study was carried out in the infection control laboratory at Assiut University Hospitals over a period of one year. Clinical samples were collected from 285 patients with various (HAIs) acquired after admission to different ICUs. Rectal swabs were taken from 14 cases for detection of enterococci carriage. In addition, 1377 environmental samples were collected from the surroundings of the patients. Identification was done by conventional bacteriological methods and confirmed by analytical profile index (API). Antimicrobial sensitivity testing was performed by Kirby Bauer disc diffusion method and detection of vancomycin resistance was done by agar screen method. For the isolates, phenotypic detection of cytolysin, gelatinase production and detection of biofilm by tube method, Congo red method and microtiter plate. We performed polymerase chain reaction (PCR) for detection of some virulence genes (gelE, cylA, vanA, vanB and esp). Results: Enterococci caused 10.5% of the HAIs. Respiratory tract infection was the predominant type (86.7%). The commonest species were E.gallinarum (36.7%), E.casseliflavus (30%), E.faecalis (30%), and E.durans (3.4 %). Vancomycin resistance was detected in a total of 40% (12/30) of those isolates. The risk factors associated with acquiring vancomycin resistant enterococci (VRE) were immune suppression (P= 0.031) and artificial feeding (P= 0.008). For the rectal swabs, enterococci species were detected in 71.4% of samples with the predominance of E. casseliflavus (50%). Most of the isolates were vancomycin resistant (70%). Out of a total 1377 environmental samples, 577 (42%) samples were contaminated with different microorganisms. Enterococci were detected in 1.7% (10/577) of total contaminated samples, 50% of which were vancomycin resistant. All isolates were resistant to penicillin, ampicillin, oxacillin, ciprofloxacin, amikacin, erythromycin, clindamycin and trimethoprim-sulfamethaxazole. For the remaining antibiotics, variable percentages of resistance were reported. Cytolysin and gelatinase were detected phenotypically in 16% and 48 % of the isolates respectively. The microtiter plate method showed the highest percentages of detection of biofilm among all isolated species (100%). The studied virulence genes gelE, esp, vanA and vanB were detected in 62%, 12%, 2% and 12% respectively, while cylA gene was not detected in any isolates. Conclusions: A significant percentage of enterococci was isolated from patients and environments in the ICUs. Many virulence factors were detected phenotypically and genotypically among isolates. The high percentage of resistance, coupled with the risk of cross transmission to other patients make enterococci infections a significant infection control issue in hospitals.

Keywords: antimicrobial resistance, enterococci, ICUs, virulence factors

Procedia PDF Downloads 265
2314 Development of a Passive Solar Tomato Dryer with Movable Heat Storage System

Authors: Jacob T. Liberty, Wilfred I. Okonkwo

Abstract:

The present study designed and constructed a post-harvest passive solar tomato dryer of dimension 176 x 152 x 54cm for drying tomato. Quality of the dried crop was evaluated and compared with the fresh ones. The solar dryer consist of solar collector (air heater), 110 x 61 x 10 x 10cm, the drying chamber, 102 x54cm, removal heat storage unit, 40 x 35 x 13cm and drying trays, 43 x 42cm. The physicochemical properties of this crop were evaluated before and after drying. Physicochemical properties evaluated includes moisture, protein, fat, fibre, ash, carbohydrate and vitamin C, contents. The fresh, open and solar dried samples were analysed for their proximate composition using the recommended method of AOAC. Also, statistical analysis of the data was conducted using analysis of variance (ANOVA) using completely Randomize Design (CRD) and means were separated by Duncan’s New Multiple Range test (DNMRT). Proximate analysis showed that solar dried tomato had significantly (P < 0.05) higher protein, fibre, ash, carbohydrate and vitamin C except for the fat content that was significantly (P < 0.05) higher for all the open sun dried samples than the solar dried and fresh product. The nutrient which is highly affected by sun drying is vitamin C. Result indicates that moisture loss in solar dried tomato was faster and lower than the open dried samples and as such makes the solar dried products of lesser tendency to mould and bacterial growth. Also, the open sun dried samples had to be carried into the sheltered place each time it rained. The solar dried produce is of high quality. Further processing of the dried crops will involve packaging for commercial purposes. This will also help in making these agricultural product available in a relatively cheap price in off season and also avert micronutrient deficiencies in diet especially among the low-income groups in Nigeria.

Keywords: tomato, passive solar dryer, physicochemical properties, removal heat storage

Procedia PDF Downloads 291
2313 Functional Analysis of Variants Implicated in Hearing Loss in a Cohort from Argentina: From Molecular Diagnosis to Pre-Clinical Research

Authors: Paula I. Buonfiglio, Carlos David Bruque, Lucia Salatino, Vanesa Lotersztein, Sebastián Menazzi, Paola Plazas, Ana Belén Elgoyhen, Viviana Dalamón

Abstract:

Hearing loss (HL) is the most prevalent sensorineural disorder affecting about 10% of the global population, with more than half due to genetic causes. About 1 in 500-1000 newborns present congenital HL. Most of the patients are non-syndromic with an autosomal recessive mode of inheritance. To date, more than 100 genes are related to HL. Therefore, the Whole-exome sequencing (WES) technique has become a cost-effective alternative approach for molecular diagnosis. Nevertheless, new challenges arise from the detection of novel variants, in particular missense changes, which can lead to a spectrum of genotype-to-phenotype correlations, which is not always straightforward. In this work, we aimed to identify the genetic causes of HL in isolated and familial cases by designing a multistep approach to analyze target genes related to hearing impairment. Moreover, we performed in silico and in vivo analyses in order to further study the effect of some of the novel variants identified in the hair cell function using the zebrafish model. A total of 650 patients were studied by Sanger Sequencing and Gap-PCR in GJB2 and GJB6 genes, respectively, diagnosing 15.5% of sporadic cases and 36% of familial ones. Overall, 50 different sequence variants were detected. Fifty of the undiagnosed patients with moderate HL were tested for deletions in STRC gene by Multiplex ligation-dependent probe amplification technique (MLPA), leading to 6% of diagnosis. After this initial screening, 50 families were selected to be analyzed by WES, achieving diagnosis in 44% of them. Half of the identified variants were novel. A missense variant in MYO6 gene detected in a family with postlingual HL was selected to be further analyzed. A protein modeling with AlphaFold2 software was performed, proving its pathogenic effect. In order to functionally validate this novel variant, a knockdown phenotype rescue assay in zebrafish was carried out. Injection of wild-type MYO6 mRNA in embryos rescued the phenotype, whereas using the mutant MYO6 mRNA (carrying c.2782C>A variant) had no effect. These results strongly suggest the deleterious effect of this variant on the mobility of stereocilia in zebrafish neuromasts, and hence on the auditory system. In the present work, we demonstrated that our algorithm is suitable for the sequential multigenic approach to HL in our cohort. These results highlight the importance of a combined strategy in order to identify candidate variants as well as the in silico and in vivo studies to analyze and prove their pathogenicity and accomplish a better understanding of the mechanisms underlying the physiopathology of the hearing impairment.

Keywords: diagnosis, genetics, hearing loss, in silico analysis, in vivo analysis, WES, zebrafish

Procedia PDF Downloads 74
2312 Resistance of Mycobacterium tuberculosis to Daptomycin

Authors: Ji-Chan Jang

Abstract:

Tuberculosis is still major health problem because there is an increase of multidrug-resistant and extensively drug-resistant forms of the disease. Therefore, the most urgent clinical need is to discover potent agents and develop novel drug combination capable of reducing the duration of MDR and XDR tuberculosis therapy. Three reference strains H37Rv, CDC1551, W-Beijing GC1237 and six clinical isolates of MDRTB were tested to daptomycin in the range of 0.013 to 256 mg/L. Daptomycin is resistant to all tested M. tuberculosis strains not only laboratory strains but also clinical MDR strains that were isolated at different source. Daptomycin will not be an antibiotic of choice for treating infection of Gram positive atypical slowly growing M. tuberculosis.

Keywords: tuberculosis, daptomycin, resistance, Mycobacterium tuberculosis

Procedia PDF Downloads 355
2311 Clinico-Microbiological Study of S. aureus from Various Clinical Samples with Reference to Methicillin Resistant S. aureus (MRSA)

Authors: T. G. Pathrikar, A. D. Urhekar, M. P. Bansal

Abstract:

To find out S. aureus from patient samples on the basis of coagulase test. We have evaluated slide coagulase (n=46 positive), tube coagulase (n=48 positive) and DNase test (n=44, positive) , We have isolated and identified MRSA from various clinical samples and specimens by disc diffusion method determined the incidence of MRSA 50% in patients. Found out the in vitro antimicrobial susceptibility pattern of MRSA isolates and also the MIC of MRSA of oxacillin by E-Test.

Keywords: cefoxitin disc diffusion MRSA detection, e – test, S. aureus devastating pathogen, tube coagulase confirmation

Procedia PDF Downloads 475
2310 Potential Hydrocarbon Degraders Present in Oil from WWII Wrecks in the Pacific

Authors: Awei Bainivalu, Joachim Larsen, Logesh Panneerselvan, Toby Mills, Brett Neilan, Megharaj Mallavarapu

Abstract:

World War II (WWII) shipwrecks harbour up to 20 million tonnes of oil. More than 3000 wrecks are in the Pacific Ocean; 300 are oil tankers. Compared to other oil removal methods, bioremediation is environmentally friendly and cost-effective. Oil's microbial community and hydrocarbon properties from the Pacific WWII wrecks were identified. Dominant phyla are Proteobacteria, Actinobacteria, and Firmicutes. Native marine bacteria oil-degraders were isolated for bioremediation. Petroleum degradation data from the bacterial consortium will be analyzed over the next three months.

Keywords: oil bioremediation, marine bacteria, WWII shipwrecks, pacific

Procedia PDF Downloads 109
2309 Extremophilic Amylases of Mycelial Fungi Strains Isolated in South Caucasus for Starch Processing

Authors: T. Urushadze, R. Khvedelidze, L. Kutateladze, M. Jobava, T. Burduli, T. Alexidze

Abstract:

There is an increasing interest in reliable, wasteless, ecologically friendly technologies. About 40% of enzymes produced all over the world are used for production of syrups with high concentration of glucose-fructose. One of such technologies complies obtaining fermentable sugar glucose from raw materials containing starch by means of amylases. In modern alcohol-producing factories this process is running in two steps, involving two enzymes of different origin: bacterial α-amylase and fungal glucoamylase, as generally fungal amylases are less thermostable as compared to bacterial amylases. Selection of stable and operable at 700С and higher temperatures enzyme preparation with both α- and glucoamylase activities will allow conducting this process in one step. S. Durmishidze Institute of Biochemistry and Biotechnology owns unique collection of mycelial fungi, isolated from different ecological niches of Caucasus. As a result of screening our collection 39 strains poducing amylases were revealed. Most of them belong to the genus Aspergillus. Optimum temperatures of action of selected amylases from three producers were estableshed to be within the range 67-80°C. A. niger B-6 showed higher α-amylase activity at 67°C, and glucoamylase activity at 62°C, A. niger 6-12 showed higher α-amylase activity at 72°C, and glucoamylase activity at 65°C, Aspergillus niger p8-3 showed higher activities at 82°C and 70°C, for α-amylase and glucoamylase activities, respectively. Exhaustive hydrolysis process of starch solutions of different concentrations (3, 5, 15, and 30 %) with cultural liquid and technical preparation of Aspergillus niger p8-3 enzyme was studied. In case of low concentrations exhaustive hydrolysis of starch lasts 40–60 minutes, in case of high concentrations hydrolysis takes longer time. 98, 6% yield of glucose can be reached at incubation during 12 hours with enzyme cultural liquid and 8 hours incubation with technical preparation of the enzyme at gradual increase of temperature from 50°C to 82°C during the first 20 minutes and further decrease of temperature to 70°C. Temperature setting for high yield of glucose and high hydrolysis (pasteurizing), optimal for activity of these strains is the prerequisite to be able to carry out hydrolysis of starch to glucose in one step, and consequently, using one strain, what will be economically justified.

Keywords: amylase, glucose hydrolisis, stability, starch

Procedia PDF Downloads 335
2308 The Interplay between Autophagy and Macrophages' Polarization in Wound Healing: A Genetic Regulatory Network Analysis

Authors: Mayada Mazher, Ahmed Moustafa, Ahmed Abdellatif

Abstract:

Background: Autophagy is a eukaryotic, highly conserved catabolic process implicated in many pathophysiologies such as wound healing. Autophagy-associated genes serve as a scaffolding platform for signal transduction of macrophage polarization during the inflammatory phase of wound healing and tissue repair process. In the current study, we report a model for the interplay between autophagy-associated genes and macrophages polarization associated genes. Methods: In silico analysis was performed on 249 autophagy-related genes retrieved from the public autophagy database and gene expression data retrieved from Gene Expression Omnibus (GEO); GSE81922 and GSE69607 microarray data macrophages polarization 199 DEGS. An integrated protein-protein interaction network was constructed for autophagy and macrophage gene sets. The gene sets were then used for GO terms pathway enrichment analysis. Common transcription factors for autophagy and macrophages' polarization were identified. Finally, microRNAs enriched in both autophagy and macrophages were predicated. Results: In silico prediction of common transcription factors in DEGs macrophages and autophagy gene sets revealed a new role for the transcription factors, HOMEZ, GABPA, ELK1 and REL, that commonly regulate macrophages associated genes: IL6,IL1M, IL1B, NOS1, SOC3 and autophagy-related genes: Atg12, Rictor, Rb1cc1, Gaparab1, Atg16l1. Conclusions: Autophagy and macrophages' polarization are interdependent cellular processes, and both autophagy-related proteins and macrophages' polarization related proteins coordinate in tissue remodelling via transcription factors and microRNAs regulatory network. The current work highlights a potential new role for transcription factors HOMEZ, GABPA, ELK1 and REL in wound healing.

Keywords: autophagy related proteins, integrated network analysis, macrophages polarization M1 and M2, tissue remodelling

Procedia PDF Downloads 131
2307 Formulation, Nutritive Value Assessment And Effect On Weight Gain Of Infant Formulae Prepared From Locally Available Materia

Authors: J. T. Johnson, R. A. Atule, E. Gbodo

Abstract:

The widespread problem of infant malnutrition in developing countries has stirred efforts in research, development and extension by both local and international organizations. As a result, the formulation and development of nutritious weaning foods from local and readily available raw materials which are cost effective has become imperative in many developing countries. Thus, local and readily available raw materials where used to compound and develop nutritious new infant formulae. The materials used for this study include maize, millet, cowpea, pumpkin, fingerlings, and fish bone. The materials where dried and blended to powder. The powders were weighed in the ratio of 4:4:4:3:1:1 respectively and were then mixed properly. Analysis of nutritive value was conducted on the formulae and compared with NAN-2 standard and results reveals that the formulae had reasonable amount of moisture, lipids, carbohydrate, protein, and fibre. Although NAN-2 was superior in both carbohydrate and protein, the new infant formula was higher in mineral elements, vitamins, fibre, and lipids. All the essentials vitamins and both macro and micro minerals where found in appreciable quantity capable of meeting the biochemical and physiological demand of the body while the anti-nutrients composition were significantly below FAO and WHO safe limits. Finally, the compounded infant formulae was feed to a set of albino Wistar rats while some other set of rats was feed with NAN-2 for the period of twenty seven (27) days and body weight was measure at three days intervals. The results of body weight changes was spectacular as their body weight over shot or almost double that of those animals that were feed with NAN-2 at each point of measurement. The results suggest that the widespread problem of infant malnutrition in the developing world especially among the low income segment of the society can now be reduced if not totally eradicated since nutritive and cost effective weaning formulae can be prepared locally from common readily available materials.

Keywords: formulation, nutritive value, local, materials

Procedia PDF Downloads 366
2306 Anti-Obesity Activity of Garcinia xanthochymus: Biochemical Characterization and In vivo Studies in High Fat Diet-Rat Model

Authors: Mahesh M. Patil, K. A. Anu-Appaiah

Abstract:

Overweight and obesity is a serious medical problem, increasing in prevalence, and affecting millions worldwide. Investigators have been trying from decades to articulate the burden of obesity and related risk factors. To answer this problem, we suggest a new therapeutic anti-obesity compounds from Garcinia xanthochymus fruit. However, there is little published scientific information on non-hydroxycitric acid Garcinia species. Our findings include biochemical characterization of the fruit; in vivo toxicity and bio-efficacy study of G. xanthochymus in high fat diet wistar rat model. We observed that Garcinia pericarp is a rich source of organic acids, polyphenols, mono- (40.63%) and poly-unsaturated fatty acids (16.45%; omega-3: 10.02%). Toxicological studies have showed that Garcinia is safe and had no observed adverse effect level up to 400 mg/kg/day. Body weight and food intake was significantly (P<0.05) reduced in oral gavage treated rats (sonicated Garcinia powder) in 13 weeks. Subcutaneous fat was significantly (P<0.05) reduced in Garcinia treated rats. Hepatocytes significantly (p<0.05) overexpressed sterol regulatory element binding protein 2, liver X receptor- α, liver X receptor- β, lipoprotein lipase and monoacylglycerol lipase. Fatty acid binding protein 1 and peroxisome proliferator activated receptor- α were down regulated as assessed by real time qPCR. Currently our research is focused on the adipocyte obesity related gene expressions, effect of Garcinia on 3T3-adipocyte cell lines and high fat diet induced mice model. This in vivo pre-clinical data suggests that G. xanthochymus may have clinical utility for the treatment of obesity. However, further studies are required to establish its potency.

Keywords: Garcinia xanthochymus, anti-obesity, high fat diet, real time qPCR

Procedia PDF Downloads 239
2305 Impact of Two Herbal Seeds Supplementation on Growth Performance and Some Biochemical Blood and Tissue Parameters of Broiler Chickens

Authors: Hamada A. Ahmed, Kadry M. Sadek, Ayman E. Taha

Abstract:

The effects of basil and/or chamomile seed supplementation on the growth of Hubbard broiler chicks were evaluated. The antioxidant effects of these supplements were also assessed. One hundred and twenty 1-day-old broiler chicks were randomly divided into four equal groups. The control group (group 1) was fed a basal diet (BD) without supplementation. Groups 2, 3, and 4 were fed the BD supplemented with 10g basil, 10g chamomile, and 5g basil plus 5g chamomile per kg of food, respectively. Basil supplementation alone or in combination with chamomile non-significantly (P≥0.05) increased final body weight (3.2% and 0.3%, respectively) and weight gain (3.5% and 3.6%, respectively) over the experimental period. Chamomile supplementation alone non-significantly (P≥0.05) reduced final body weight and weight gain over the experimental period by 1.7% and 1.7%, respectively. In comparison to the control group, herbal seed supplementation reduced feed intake and improved the feed conversion and protein efficiency ratios. In general, basil seed supplementation stimulated chicken growth and improved the feed efficiency more effectively than chamomile seed supplementation. The antioxidant activities of basil and/or chamomile supplementation were examined in the thymus, bursa, and spleen. In chickens that received supplements, the level of malondialdehyde was significantly decreased, whereas the activities of glutathione, superoxide dismutase, and catalase were significantly increased (P<0.05). Supplementation of basil and/or chamomile did not affect blood protein levels, but had lipid-lowering effects as evidenced by reduced serum levels of total lipids, triglycerides, and cholesterol. In conclusion, supplementation of basil and/or chamomile improved growth parameters in broiler chicks and had antioxidant and blood lipid-lowering effects. These beneficial effects of basil and/or chamomile supplementation resulted in economically viable production of high-quality white meat containing no harmful residues.

Keywords: herbal additives, basil, chamomile, broiler, growth performance, antioxidant

Procedia PDF Downloads 527
2304 Heamatological and Biochemical Changes in Cockerels Fed Graded Levels of Wild Sunflower Leaf Meal

Authors: Siyanbola Mojisola Funmilayo, Amao Emmanuel Ayodele

Abstract:

The poultry industry in Nigeria has been played by a variety of problems, which include the search for feed ingredients that are not competed for by man. This has resulted in a reduced interest of farmers in the industry leading to a reduction in animal protein availability for human consumption as a consequence of a high cost of production. The incorporation of wild sunflower meal (Tithonia diversfolia, Hemsl A. Gray) (WSF Meal) and some others in poultry diets have been reported to result in compounded feed with nutrient profiles that compare favourable with feeds of conventional feedstuff and reduce feed cost as they reduce competition with humans. A 98-day feeding trial was used to evaluate the effect of Wild sunflower leaf (WSL) at varying levels on the hematology and biochemistry of cockerels. A total of one hundred and twenty(120) cockerel birds were randomly allotted into four experimental diets with three replicates per experimental diet (ten birds per replicate). Wild sunflower leaf was included in four graded levels ; 0, 5, 10, and 15%. Packed cell volume, Red blood cell count, White blood cell count, Hemoglobin count, Lymphocyte count, Neutrophil count, Platelets, Mean Corpuscular Hemoglobin Concentration (MCHC), Mean Corpuscular Hemoglobin (MCH), Aspartate aminotransferase (AST), Glucose, Urea, Chloride, Sodium, and Potassium ion values were significantly different (p<0.05) among the treatments. Mean values obtained for Creatinine, Total Protein, Alanine aminotransferase (ALT), Albumin, and Mean Corpuscular Volume (MCV) were not significantly different (p>0.05) in all the treatment. WSL could be included up to 15% in the diet of cockerel without any deleterious effect on the birds. Based on the results, up to 15% Wild sunflower meal (WSL) can be included in the diet of cockerel without any adverse effect on the hematology and biochemical indices of birds.

Keywords: biochemical changes, cockerels, hematology, wild sunflower leaf

Procedia PDF Downloads 429