Search results for: chicken protein hydrolysate
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 2544

Search results for: chicken protein hydrolysate

684 Therapeutical Role of Copper Oxide Nanoparticles (CuO NPs) for Breast Cancer Therapy

Authors: Dipranjan Laha, Parimal Karmakar

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Metal oxide nanoparticles are well known to generate oxidative stress and deregulate normal cellular activities. Among these, transition metals copper oxide nanoparticles (CuO NPs) are more compelling than others and able to modulate different cellular responses. In this work, we have synthesized and characterized CuO NPs by various biophysical methods. These CuO NPs (~30 nm) induce autophagy in human breast cancer cell line, MCF7 in a time and dose-dependent manner. Cellular autophagy was tested by MDC staining, induction of green fluorescent protein light chain 3 (GFP-LC3B) foci by confocal microscopy, transfection of pBABE-puro mCherry-EGFP-LC3B plasmid and western blotting of autophagy marker proteins LC3B, beclin1, and ATG5. Further, inhibition of autophagy by 3-Methyladenine (3-MA) decreased LD50 doses of CuO NPs. Such cell death was associated with the induction of apoptosis as revealed by FACS analysis, cleavage of PARP, dephosphorylation of Bad and increased cleavage product of caspase3. siRNA-mediated inhibition of autophagy-related gene beclin1 also demonstrated similar results. Finally, induction of apoptosis by 3-MA in CuO NPs treated cells were observed by TEM. This study indicates that CuO NPs are a potent inducer of autophagy which may be a cellular defense against the CuO NPs mediated toxicity and inhibition of autophagy switches the cellular response into apoptosis. A combination of CuO NPs with the autophagy inhibitor is essential to induce apoptosis in breast cancer cells. Acknowledgments: The authors would like to acknowledge for financial support for this research work to the Department of Biotechnology (No. BT/PR14661/NNT/28/494/2010), Government of India.

Keywords: nanoparticle, autophagy, apoptosis, siRNA-mediated inhibition

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683 Encapsulation of Probiotic Bacteria in Complex Coacervates

Authors: L. A. Bosnea, T. Moschakis, C. Biliaderis

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Two probiotic strains of Lactobacillus paracasei subsp. paracasei (E6) and Lactobacillus paraplantarum (B1), isolated from traditional Greek dairy products, were microencapsulated by complex coacervation using whey protein isolate (WPI, 3% w/v) and gum arabic (GA, 3% w/v) solutions mixed at different polymer ratio (1:1, 2:1 and 4:1). The effect of total biopolymer concentration on cell viability was assessed using WPI and GA solutions of 1, 3 and 6% w/v at a constant ratio of 2:1. Also, several parameters were examined for optimization of the microcapsule formation, such as inoculum concentration and the effect of ionic strength. The viability of the bacterial cells during heat treatment and under simulated gut conditions was also evaluated. Among the different WPI/GA weight ratios tested (1:1, 2:1, and 4:1), the highest survival rate was observed for the coacervate structures made with the ratio of 2:1. The protection efficiency at low pH values is influenced by both concentration and the ratio of the added biopolymers. Moreover, the inoculum concentration seems to affect the efficiency of microcapsules to entrap the bacterial cells since an optimum level was noted at less than 8 log cfu/ml. Generally, entrapment of lactobacilli in the complex coacervate structure enhanced the viability of the microorganisms when exposed to a low pH environment (pH 2.0). Both encapsulated strains retained high viability in simulated gastric juice (>73%), especially in comparison with non-encapsulated (free) cells (<19%). The encapsulated lactobacilli also exhibited enhanced viability after 10–30 min of heat treatment (65oC) as well as at different NaCl concentrations (pH 4.0). Overall, the results of this study suggest that complex coacervation with WPI/GA has a potential to deliver live probiotics in low pH food systems and fermented dairy products; the complexes can dissolve at pH 7.0 (gut environment), releasing the microbial cells.

Keywords: probiotic, complex coacervation, whey, encapsulation

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682 Effect of Miconazole Nitrate on Immunological Response and Its Preventive Efficacy in Labeo rohita Fingerlings against Oomycetes Saprolegnia parasitica

Authors: Mukta Singh, Ratan Kumar Saha, Himadri Saha, Paramveer Singh

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The present study evaluated the effect of sub-lethal doses of antifungal drug miconazole nitrate (MCZ) on immunological responses including immune-related gene expression and its role as a prophylactic drug against S. parasitica in Labeo rohita fingerlings. Fish were fed with sub lethal doses of MCZ i.e., T1- 6.30 mg MCZ kgBW⁻¹, T2- 12.61 mg MCZ kgBW⁻¹ and T3- 25.22 mg MCZ kgBW⁻¹ and sampling was done at different time intervals for 240 h. Immunological parameters viz. lysozyme activity, oxygen radical production and plasma anti-protease activity showed significant enhancement (p < 0.05) in fish fed with T2 and T3 doses. Significant reduction in plasma protein content was observed in all the dietary groups as compared to control. Expression of immune-relevant genes like TLR-22 and β2-M showed significantly higher expression at six h and 24 h of sampling in both liver and head-kidney. However, these genes showed a down-regulation after 120 h of sampling in both the tissues. Preventive efficacy study showed that single dose of MCZ provides protection against oomycetes up to the fourth day of infection. Significantly higher mortality was observed in control diet-fed fish as compared to fish fed with MCZ medicated diet. Thus, from the study, it can be concluded that the MCZ can act as a potent antifungal agent for preventing oomycetes infection as well as to enhance the immune response.

Keywords: antifungal, immune gene, immunological, miconazole nitrate, prophylactic

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681 WT1 Exprassion in Malignant Surface Epithelial Ovarian Tumors

Authors: Mahmoodreza Tahamtan

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Background: Malignant surface epithelial ovarian tumors (SEOT) account for approximately 90% of primary ovarian cancer. Wilms tumor gene (WT1) product was defined as a tumor suppressor gene, but today it is considered capable of performing oncogenic functions. There seems to be differences in WT1 expression patterns among SEOT subtypes. We evaluate the immunohistochemical expression of WT1 protein among different histologic subtypes of SEOT. Materials and Methods: Immunohistochemistry for WT1 was done on 35 serous cystadenocarcinomas, 9 borderline serous tumors, 3 mucinous cystadenocarcinomas, 10 borderline mucinous tumors, 7 endometrioid ovarian carcinomas, 3 clear cell carcinomas, 1 malignant Brenner tumor, 2 metastatic adenocarcinomas, and 6 endometrial adenocarcinomas. A tumor was considered negative if < 1% of tumor cells were stained.Positive reactions were graded as follows:1+,1%-24%; 2+,25%-49%; 3+,50%-74%; 4+,75%-100%. Results: Of the 35 cases of ovarian serous cystadenocarcinoma, 30(85.7%) were diffusely positive (3+,4+),4 showed reactivity of < 50% of the tumor cells (1+,2+), and one were negative. All 9 borderline serous tumors showed immunoreactivity with WT1. All the mucinous tumors(n:13), endometrioid carcinomas (n: 7), clear cell carcinomas (n: 3), metastatic adenocarcinomas (n: 2) and primary endometrial carcinomas (n:6) were negative. The single malignant Brenner tumor showed a positive reaction for WT1(4+) Conclusion: WT1 is a good marker to distinguish primary ovarian serous carcinomas from other surface epithelial tumors (especially endometrioid subtype) and metastatic carcinomas (especially endometrial serous carcinoma), other than malignant mesothelioma. We cannot rely to the degree of expression inorder to separate high grade borderline serous tumors from low grade ones.

Keywords: WT1, ovary, epithelial tumors, malignant

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680 Level of IGF-I and IGFBP-3 in Gingival Crevicular Fluid and Plasma in Patients with Aggressive Periodontitis

Authors: Youjeong Hwang

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Purpose: Insulin-like growth factor-I (IGF-I) promotes B-cell development, immunoglobulin formation, and interleukin-6 (IL-6) production, then regulate the immune response and inflammation. As IGF-I and their receptor also exist in the periodontal tissue, they may affect the immune response caused by periodontal pathogens in aggressive periodontitis (AgP) patients. The function of IGF is regulated by IGF binding proteins (IGFBPs), and IGFBP-3 is known to most abundant in plasma. The aim of the present study was to assess the concentration of IGF-I and IGFBP-3 in plasma and gingival crevicular fluid (GCF) in AgP patients and to find out their association. Methods: Nine patients with AgP (test group) and nine healthy subjects (control group) were included in this study. None of the subjects had a history of systemic disease, smoking or steroids medication. GCF samples were collected by microcapillary pipettes and plasma samples were obtained by venipuncture. Probing pocket depth (PD), clinical attachment level (CAL) and bleeding on probing (BOP) were recorded. Samples were assayed for IGF-I and IGFBP-3 levels using ELISA. Results: Mean IGF-I level in GCF was higher in the test group than control. Mean IGF-I level in plasma and IGFBP-3 level in GCF and plasma in control group were higher than that of the test group. However, there was no statistical significance (p > 0.05). The mean level of IGF-I and IGFBP-3 in GCF was lower than those in plasma. Mean IGF-I level in plasma showed a negative correlation with PD and CAL (p < 0.05) in both groups. The levels of IGF-I and IGFBP-3 in GCF seemed to be negatively correlated with BOP in the test group (p < 0.05). Conclusions: The difference in the level of IGF-I and IGFBP-3 between AgP and healthy subjects was not significant. Further studies that explain the mechanism of the protective role of IGF-I with more samples are needed.

Keywords: aggressive periodontitis, pathogenesis, insulin-like growth factor, insulin-like growth factor binding protein

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679 Nutritional Status of Morbidly Obese Patients Prior to Bariatric Surgery

Authors: Azadeh Mottaghi, Reyhaneh Yousefi, Saeed Safari

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Background: Bariatric surgery is widely proposed as the most effective approach to mitigate the growing pace of morbid obesity. As bariatric surgery candidates suffer from pre-existing nutritional deficiencies, it is of great importance to assess nutritional status of candidates before surgery in order to establish appropriate nutritional interventions. Objectives: The present study assessed and represented baseline data according to the nutritional status among candidates for bariatric surgery. Methods: A cross-sectional analysis of pre-surgery data was collected on 170 morbidly obese patients undergoing bariatric surgery between October 2017 and February 2018. Dietary intake data (evaluated through 147-item food frequency questionnaire), anthropometric measures and biochemical parameters were assessed. Results: Participants included 145 females (25 males) with average age of 37.3 ± 10.2 years, BMI of 45.7 ± 6.4 kg/m² and reported to have a total of 72.3 ± 22.2 kg excess body weight. The most common nutritional deficiencies referred to iron, ferritin, transferrin, albumin, vitamin B12, and vitamin D, the prevalence of which in the study population were as followed; 6.5, 6.5, 3, 2, 17.6 and 66%, respectively. Mean energy, protein, fat, and carbohydrate intake were 3887.3 ± 1748.32 kcal/day, 121.6 ± 57.1, 144.1 ± 83.05, and 552.4 ± 240.5 gr/day, respectively. The study population consumed lower levels of iron, calcium, folic acid, and vitamin B12 compared to the Dietary Reference Intake (DRI) recommendations (2, 26, 2.5, and 13%, respectively). Conclusion: According to the poor dietary quality of bariatric surgery candidates, leading to nutritional deficiencies pre-operatively, close monitoring and tailored supplementation pre- and post-bariatric surgery are required.

Keywords: bariatric surgery, food frequency questionnaire, obesity, nutritional status

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678 Collagen Scaffold Incorporated with Macrotyloma uniflorum Plant Extracts as a–Burn/Wound Dressing Material, in Vitro and in Vivo Evaluation

Authors: Thangavelu Muthukumar, Thotapalli Parvathaleswara Sastry

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Collagen is the most abundantly available connective tissue protein, which is being used as a biomaterial for various biomedical applications. Presently, fish wastes are disposed improperly which is causing serious environmental pollution resulting in offensive odour. Fish scales are promising source of Type I collagen. Medicinal plants have been used since time immemorial for treatment of various ailments of skin and dermatological disorders especially cuts, wounds, and burns. Developing biomaterials from the natural sources which are having wound healing properties within the search of a common man is the need of hour, particularly in developing and third world countries. With these objectives in view we have developed a wound dressing material containing fish scale collagen (FSC) incorporated with Macrotyloma uniflorum plant extract (PE). The wound dressing composite was characterized for its physiochemical properties using conventional methods. SEM image revealed that the composite has fibrous and porous surface which helps in transportation of oxygen as well as absorbing wound fluids. The biomaterial has shown 95% biocompatibility with required mechanical strength and has exhibited antimicrobial properties. This biomaterial has been used as a wound dressing material in experimental wounds of rats. The healing pattern was evaluated by macroscopic observations, panimetric studies, biochemical, histopathological observations. The results showed faster healing pattern in the wounds treated with CSPE compared to the other composites used in this study and untreated control. These experiments clearly suggest that CSPE can be used as wound/burn dressing materials.

Keywords: collagen, wound dressing, Macrotyloma uniflorum, burn dressing

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677 Hippocampus Proteomic of Major Depression and Antidepressant Treatment: Involvement of Cell Proliferation, Differentiation, and Connectivity

Authors: Dhruv J. Limaye, Hanga Galfalvy, Cheick A. Sissoko, Yung-yu Huang, Chunanning Tang, Ying Liu, Shu-Chi Hsiung, Andrew J. Dwork, Gorazd B. Rosoklija, Victoria Arango, Lewis Brown, J. John Mann, Maura Boldrini

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Memory and emotion require hippocampal cell viability and connectivity and are disrupted in major depressive disorder (MDD). Applying shotgun proteomics and stereological quantification of neural progenitor cells (NPCs), intermediate neural progenitors (INPs), and mature granule neurons (GNs), to postmortem human hippocampus, identified differentially expressed proteins (DEPs), and fewer NPCs, INPs and GNs, in untreated MDD (uMDD) compared with non-psychiatric controls (CTRL) and antidepressant-treated MDD (MDDT). DEPs lower in uMDD vs. CTRL promote mitosis, differentiation, and prevent apoptosis. DEPs higher in uMDD vs. CTRL inhibit the cell cycle, and regulate cell adhesion, neurite outgrowth, and DNA repair. DEPs lower in MDDT vs. uMDD block cell proliferation. We observe group-specific correlations between numbers of NPCs, INPs, and GNs and an abundance of proteins regulating mitosis, differentiation, and apoptosis. Altered protein expression underlies hippocampus cellular and volume loss in uMDD, supports a trophic effect of antidepressants, and offers new treatment targets.

Keywords: proteomics, hippocampus, depression, mitosis, migration, differentiation, mitochondria, apoptosis, antidepressants, human brain

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676 Organotin (IV) Based Complexes as Promiscuous Antibacterials: Synthesis in vitro, in Silico Pharmacokinetic, and Docking Studies

Authors: Wajid Rehman, Sirajul Haq, Bakhtiar Muhammad, Syed Fahad Hassan, Amin Badshah, Muhammad Waseem, Fazal Rahim, Obaid-Ur-Rahman Abid, Farzana Latif Ansari, Umer Rashid

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Five novel triorganotin (IV) compounds have been synthesized and characterized. The tin atom is penta-coordinated to assume trigonal-bipyramidal geometry. Using in silico derived parameters; the objective of our study is to design and synthesize promiscuous antibacterials potent enough to combat resistance. Among various synthesized organotin (IV) complexes, compound 5 was found as potent antibacterial agent against various bacterial strains. Further lead optimization of drug-like properties was evaluated through in silico predictions. Data mining and computational analysis were utilized to derive compound promiscuity phenomenon to avoid drug attrition rate in designing antibacterials. Xanthine oxidase and human glucose- 6-phosphatase were found as only true positive off-target hits by ChEMBL database and others utilizing similarity ensemble approach. Propensity towards a-3 receptor, human macrophage migration factor and thiazolidinedione were found as false positive off targets with E-value 1/4> 10^-4 for compound 1, 3, and 4. Further, displaying positive drug-drug interaction of compound 1 as uricosuric was validated by all databases and docked protein targets with sequence similarity and compositional matrix alignment via BLAST software. Promiscuity of the compound 5 was further confirmed by in silico binding to different antibacterial targets.

Keywords: antibacterial activity, drug promiscuity, ADMET prediction, metallo-pharmaceutical, antimicrobial resistance

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675 The Production of Collagen and Collagen Peptides from Nile Tilapia Skin Using Membrane Technology

Authors: M. Thuanthong, W. Youravong, N. Sirinupong

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Nile tilapia (Oreochromis niloticus) is one of fish species cultured in Thailand with a high production volume. A lot of skin is generated during fish processing. In addition, there are many research reported that fish skin contains abundant of collagen. Thus, the use of Nile tilapia skin as collagen source can increase the benefit of industrial waste. In this study, Acid soluble collagen (ASC) was extracted at 5, 15 or 25 ˚C with 0.5 M acetic acid then the acid was removed out and collagen was concentrated by ultrafiltration-diafiltration (UFDF). The triple helix collagen from UFDF process was used as substrate to produce collagen peptides by alcalase hydrolysis in an enzymatic membrane reactor (EMR) coupling with 1 kDa molecular weight cut off (MWCO) polysulfone hollow fiber membrane. The results showed that ASC extracted at high temperature (25 ˚C) with 0.5 M acetic acid for 5 h still preserved triple helix structure. In the UFDF process, the acid removal was higher than 90 % without any effect on ASC properties, particularly triple helix structure as indicated by circular dichroism spectrum. Moreover, Collagen from UFDF was used to produce collagen peptides by EMR. In EMR, collagen was pre-hydrolyzed by alcalase for 60 min before introduced to membrane separation. The EMR operation was operated for 10 h and provided a good of protein conversion stability. The results suggested that there is a successfulness of UF in application for acid removal to produce ASC with desirable preservation of its quality. In addition, the EMR was proven to be an effective process to produce low molecular weight peptides with ACE-inhibitory activity properties.

Keywords: acid soluble collagen, ultrafiltration-diafiltration, enzymatic membrane reactor, ace-inhibitory activity

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674 Hybrid-Nanoengineering™: A New Platform for Nanomedicine

Authors: Mewa Singh

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Nanomedicine, a fusion of nanotechnology and medicine, is an emerging technology ideally suited to the targeted therapies. Nanoparticles overcome the low selectivity of anti-cancer drugs toward the tumor as compared to normal tissue and hence result-in less severe side-effects. Our new technology, HYBRID-NANOENGINEERING™, uses a new molecule (MR007) in the creation of nanoparticles that not only helps in nanonizing the medicine but also provides synergy to the medicine. The simplified manufacturing process will result in reduced manufacturing costs. Treatment is made more convenient because hybrid nanomedicines can be produced in oral, injectable or transdermal formulations. The manufacturing process uses no protein, oil or detergents. The particle size is below 180 nm with a narrow distribution of size. Importantly, these properties confer great stability of the structure. The formulation does not aggregate in plasma and is stable over a wide range of pH. The final hybrid formulation is stable for at least 18 months as a powder. More than 97 drugs, including paclitaxel, docetaxel, tamoxifen, doxorubicinm prednisone, and artemisinin have been nanonized in water soluble formulations. Preclinical studies on cell cultures of tumors show promising results. Our HYBRID-NANOENGINEERING™ platform enables the design and development of hybrid nano-pharmaceuticals that combine efficacy with tolerability, giving patients hope for both extended overall survival and improved quality of life. This study would discuss or present this new discovery of HYBRID-NANOENGINEERING™ which targets drug delivery, synergistic, and potentiating effects, and barriers of drug delivery and advanced drug delivery systems.

Keywords: nano-medicine, nano-particles, drug delivery system, pharmaceuticals

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673 hsa-miR-1204 and hsa-miR-639 Prominent Role in Tamoxifen's Molecular Mechanisms on the EMT Phenomenon in Breast Cancer Patients

Authors: Mahsa Taghavi

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In the treatment of breast cancer, tamoxifen is a regularly prescribed medication. The effect of tamoxifen on breast cancer patients' EMT pathways was studied. In this study to see if it had any effect on the cancer cells' resistance to tamoxifen and to look for specific miRNAs associated with EMT. In this work, we used continuous and integrated bioinformatics analysis to choose the optimal GEO datasets. Once we had sorted the gene expression profile, we looked at the mechanism of signaling, the ontology of genes, and the protein interaction of each gene. In the end, we used the GEPIA database to confirm the candidate genes. after that, I investigated critical miRNAs related to candidate genes. There were two gene expression profiles that were categorized into two distinct groups. Using the expression profile of genes that were lowered in the EMT pathway, the first group was examined. The second group represented the polar opposite of the first. A total of 253 genes from the first group and 302 genes from the second group were found to be common. Several genes in the first category were linked to cell death, focal adhesion, and cellular aging. Two genes in the second group were linked to cell death, focal adhesion, and cellular aging. distinct cell cycle stages were observed. Finally, proteins such as MYLK, SOCS3, and STAT5B from the first group and BIRC5, PLK1, and RAPGAP1 from the second group were selected as potential candidates linked to tamoxifen's influence on the EMT pathway. hsa-miR-1204 and hsa-miR-639 have a very close relationship with the candidates genes according to the node degrees and betweenness index. With this, the action of tamoxifen on the EMT pathway was better understood. It's important to learn more about how tamoxifen's target genes and proteins work so that we can better understand the drug.

Keywords: tamoxifen, breast cancer, bioinformatics analysis, EMT, miRNAs

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672 Biophysical Consideration in the Interaction of Biological Cell Membranes with Virus Nanofilaments

Authors: Samaneh Farokhirad, Fatemeh Ahmadpoor

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Biological membranes are constantly in contact with various filamentous soft nanostructures that either reside on their surface or are being transported between the cell and its environment. In particular, viral infections are determined by the interaction of viruses (such as filovirus) with cell membranes, membrane protein organization (such as cytoskeletal proteins and actin filament bundles) has been proposed to influence the mechanical properties of lipid membranes, and the adhesion of filamentous nanoparticles influence their delivery yield into target cells or tissues. The goal of this research is to integrate the rapidly increasing but still fragmented experimental observations on the adhesion and self-assembly of nanofilaments (including filoviruses, actin filaments, as well as natural and synthetic nanofilaments) on cell membranes into a general, rigorous, and unified knowledge framework. The global outbreak of the coronavirus disease in 2020, which has persisted for over three years, highlights the crucial role that nanofilamentbased delivery systems play in human health. This work will unravel the role of a unique property of all cell membranes, namely flexoelectricity, and the significance of nanofilaments’ flexibility in the adhesion and self-assembly of nanofilaments on cell membranes. This will be achieved utilizing a set of continuum mechanics, statistical mechanics, and molecular dynamics and Monte Carlo simulations. The findings will help address the societal needs to understand biophysical principles that govern the attachment of filoviruses and flexible nanofilaments onto the living cells and provide guidance on the development of nanofilament-based vaccines for a range of diseases, including infectious diseases and cancer.

Keywords: virus nanofilaments, cell mechanics, computational biophysics, statistical mechanics

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671 Development of Standard Thai Appetizer in Rattanakosin Era‘s Standard: Case Study of Thai Steamed Dumpling

Authors: Nunyong Fuengkajornfung, Pattama Hirunyophat, Tidarat Sanphom

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The objectives of this research were: To study of the recipe standard of Thai steamed dumpling, to study the ratio of modified starch in Thai steamed dumpling, to study chemical elements analyzing and Escherichia coli in Thai steamed dumpling. The experimental processes were designed in two stages as follows: To study the recipe standard of Thai steamed dumpling and to study the ratio of rice flour: modify starch by three levels 90:10, 73:30, and 50:50. The evaluation test used 9 Points Hedonic Scale method by the sensory evaluation test such as color, smell, taste, texture and overall liking. An experimental by Randomized Complete Block Design (RCBD). The statistics used in data analyses were means, standard deviation, one-way ANOVA and Duncan’s New Multiple Range Test. Regression equation, at a statistically significant level of .05. The results showed that the recipe standard was studied from three recipes by the sensory evaluation test such as color, odor, taste, spicy, texture and total acceptance. The result showed that the recipe standard of second was suitably to development. The ratio of rice flour: modified starch had 3 levels 90:10, 73:30, and 50:50 which the process condition of 50:50 had well scores (like moderately to like very much; used 9 Points Hedonic Scale method for the sensory test). Chemical elements analyzing, it showed that moisture 58.63%, fat 5.45%, protein 4.35%, carbohydrate 30.45%, and Ash 1.12%. The Escherichia coli is not found in lab testing.

Keywords: Thai snack in Rattanakosin era, Thai steamed dumpling, modify starch, recipe standard

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670 Biodegradable and Bioactive Scaffold for Bone Tissue Engineering

Authors: A. M. Malagon Escandon, J. A. Arenas Alatorre, C. P. Chaires Rosas, N. A. Vazquez Torres, B. Hernandez Tellez, G. Pinon Zarate, M. Herrera Enriquez, A. E. Castell Rodriguez

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The current approach to the treatment of bone defects involves the use of scaffolds that provide a biological and mechanically stable niche to favor tissue repair. Despite the significant progress in the field of bone tissue engineering, several main problems associated are attributed to giving a low biodegradation degree, does not promote osseointegration and regeneration, if the bone is not healing as well as expected or fails to heal, will not be given a proper ossification or new bone formation. The actual approaches of bone tissue regeneration are directed to the use of decellularized native extracellular matrices, which are able of retain their own architecture, mechanic properties, biodegradability and promote new bone formation because they are capable of conserving proteins and other factors that are founded in physiological concentrations. Therefore, we propose an extracellular matrix-based bioscaffolds derived from bovine cancellous bone, which is processed by decellularization, demineralization, and hydrolysis of the collagen protein, these protocols have been successfully carried out in other organs and tissues; the effectiveness of its biosafety has also been previously evaluated in vivo and Food and Drug Administration (FDA) approved. In the specific case of bone, a more complex treatment is needed in comparison with other organs and tissues because is necessary demineralization and collagen denaturalization. The present work was made in order to obtain a temporal scaffold that succeed in degradation in an inversely proportional way to the synthesis of extracellular matrix and the maturation of the bone by the cells of the host.

Keywords: bioactive, biodegradable, bone, extracellular matrix-based bioscaffolds, stem cells, tissue engineering

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669 Assessment of Breeding Soundness by Comparative Radiography and Ultrasonography of Rabbit Testes

Authors: Adenike O. Olatunji-Akioye, Emmanual B Farayola

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In order to improve the animal protein recommended daily intake of Nigerians, there is an upsurge in breeding of hitherto shunned food animals one of which is the rabbit. Radiography and ultrasonography are tools for diagnosing disease and evaluating the anatomical architecture of parts of the body non-invasively. As the rabbit is becoming a more important food animal, to achieve improved breeding of these animals, the best of the species form a breeding stock and will usually depend on breeding soundness which may be evaluated by assessment of the male reproductive organs by these tools. Four male intact rabbits weighing between 1.2 to 1.5 kg were acquired and acclimatized for 2 weeks. Dorsoventral views of the testes were acquired using a digital radiographic machine and a 5 MHz portable ultrasound scanner was used to acquire images of the testes in longitudinal, sagittal and transverse planes. Radiographic images acquired revealed soft tissue images of the testes in all rabbits. The testes lie in individual scrotal sacs sides on both sides of the midline at the level of the caudal vertebrae and thus are superimposed by caudal vertebrae and the caudal limits of the pelvic girdle. The ultrasonographic images revealed mostly homogenously hypoechogenic testes and a hyperechogenic mediastinum testis. The dorsal and ventral poles of the testes were heterogeneously hypoechogenic and correspond to the epididymis and spermatic cord. The rabbit is unique in the ability to retract the testes particularly when stressed and so careful and stressless handling during the procedures is of paramount importance. The imaging of rabbit testes can be safely done using both imaging methods but ultrasonography is a better method of assessment and evaluation of soundness for breeding.

Keywords: breeding soundness, rabbit, radiography, ultrasonography

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668 Design and Synthesis of Some Oxadiazole Bearing Benzimidazole Derivatives as Potential Epidermal Growth Factor Receptor Inhibitors

Authors: Ismail Celik, Gulgun Ayhan Kilcigil, Berna Guven, Zumra Kara, Arzu Onay-Besikci

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Epidermal Growth Factor Receptor is the cell-surface receptor of the ErbB (erythroblastic leukemia viral oncogene homologue receptors) family of tyrosine kinases. It plays a vital role in regulating the proliferation and differentiation of cells. However, a variety of mechanisms, such as EGFR expression, mutation, and ligand-dependent receptor dimerization, are associated with the development of various activated EGFR tumors. EGFR is highly expressed in most solid tumors, including breast, head and neck cancer, non-small cell lung cancer (NSCLC), renal, ovarian, and colon cancers. Thus, specific EGFR inhibition plays one of the key roles in cancer treatment. The compounds used in the treatment as tyrosine kinase inhibitors are known to contain the benzimidazole isosterium indole, pazopanib, and axitinibin indazole rings. In addition, benzimidazoles have been shown to exhibit protein kinase inhibitory activity in addition to their different biological activities.Based on these data, it was planned and synthesized of some oxadiazole bearing benzimidazole derivatives [N-cyclohexyl-5-((2-phenyl/substitutedphenyl-1H-benzo[d]imidazole-1-yl) methyl)-1,3,4-oxadiazole-2-amine]. EGFR kinase inhibitory efficiency of the synthesized compounds was determined by comparing them with a known kinase inhibitor erlotinib in vitro, and two of the compounds bearing phenyl (19a) and 3,4-dibenzyloxyphenyl (21a) ring exhibited significant activities.

Keywords: benzimidazole, EGFR kinase inhibitory, oxadiazole, synthesis

Procedia PDF Downloads 139
667 Value Addition of Quinoa (Chenopodium Quinoa Willd.) Using an Indigenously Developed Saponin Removal Machine

Authors: M.A. Ali, M. Matloob, A. Sahar, M. Yamin, M. Imran, Y.A. Yusof

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Quinoa (Chenopodium quinoa Willd.) is known as pseudocereal was originated in South America's Andes. Quinoa is a good source of protein, amino acids, micronutrients and bioactive components. The lack of gluten makes it suitable for celiac patients. Saponins, the leading ant-nutrient, are found in the pericarp, which adheres to the seed and transmits the bitter flavor to the quinoa grain. It is found in varying amounts in quinoa from 0.1% to 5%. This study was planned to design an indigenous machine to remove saponin from quinoa grains at the farm level to promote entrepreneurship. The machine consisted of a feeding hopper, rotating shaft, grooved stone, perforated steel cylinder, V-belts, pulleys, electric motor and mild steel angle iron and sheets. The motor transmitted power to the shaft with a belt drive. The shaft on which the grooved stone was attached rotated inside the perforated cylinder having a clearance of 2 mm and was removed saponin by an abrasion mechanism. The saponin-removed quinoa was then dipped in water to determine the presence of saponin as it produced foam in water and data were statistically analyzed. The results showed that the raw seed feeding rate of 25 g/s and milling time of 135 s completely removed saponin from seeds with minimum grain losses of 2.85% as compared to the economic analysis of the machine showed that its break-even point was achieved after one and half months with 18,000 s and a production capacity of 33 g/s.

Keywords: quinoa seeds, saponin, abrasion mechanism, stone polishing, indigenous machine

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666 The Effect of Electrical Discharge Plasma on Inactivation of Escherichia Coli MG 1655 in Pure Culture

Authors: Zoran Herceg, Višnja Stulić, Anet Režek Jambrak, Tomislava Vukušić

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Electrical discharge plasma is a new non-thermal processing technique which is used for the inactivation of contaminating and hazardous microbes in liquids. Plasma is a source of different antimicrobial species including UV photons, charged particles, and reactive species such as superoxide, hydroxyl radicals, nitric oxide and ozone. Escherichia coli was studied as foodborne pathogen. The aim of this work was to examine inactivation effects of electrical discharge plasma treatment on the Escherichia coli MG 1655 in pure culture. Two types of plasma configuration and polarity were used. First configuration was with titanium wire as high voltage needle and another with medical stainless steel needle used to form bubbles in treated volume and titanium wire as high voltage needle. Model solution samples were inoculated with Escerichia coli MG 1655 and treated by electrical discharge plasma at treatment time of 5 and 10 min, and frequency of 60, 90 and 120 Hz. With the first configuration after 5 minutes of treatment at frequency of 120 Hz the inactivation rate was 1.3 log₁₀ reduction and after 10 minutes of treatment the inactivation rate was 3.0 log₁₀ reduction. At the frequency of 90 Hz after 10 minutes inactivation rate was 1.3 log₁₀ reduction. With the second configuration after 5 minutes of treatment at frequency of 120 Hz the inactivation rate was 1.2 log₁₀ reduction and after 10 minutes of treatment the inactivation rate was also 3.0 log₁₀ reduction. In this work it was also examined the formation of biofilm, nucleotide and protein leakage at 260/280 nm, before and after treatment and recuperation of treated samples. Further optimization of method is needed to understand mechanism of inactivation.

Keywords: electrical discharge plasma, escherichia coli MG 1655, inactivation, point-to-plate electrode configuration

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665 Effects of Exhaust Gas Emitted by the Fleet on Public Health in the Region of Annaba (Algeria): Ecotoxicological Test on Durum Wheat (Triticum durum Desf.)

Authors: Aouissi Nora, Meksem Leila

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This work focused on the study of air pollution generated by the transport sector in the region of Annaba. Our study is based on two parts: the first one concerns an epidemiological investigation in the area of Annaba situated in the east Algerian coast, which deals with the development of the fleet and its impact on public health. To get a more precise idea of the impact of road traffic on public health, we consulted the computing center office of the National Social Insurance Fund. The information we were given by this office refers to the number of reported asthma and heart disease after medical examination during the period 2006-2010. The second part was devoted to the study of the toxicity of exhaust gases on some physical and biochemical parameters of durum wheat (Triticum durum Desf.). After germination and three-leaf stage, the pots are placed in a box of volume (0,096 m3) having an input which is linked directly to the exhaust pipe of a truck, and an outlet to prevent asphyxiation plant. The experience deals with 30 pots: 10 pots are exposed for 5 minutes to exhaust smoke; the other 10 are exposed for 15 minutes, and the remaining 10 for 30 minutes. The epidemiological study shows that the levels of pollutants emitted by the fleet are responsible for the increase of people respiratory and cardiovascular diseases. As for biochemical analyses of vegetation, they clearly show the toxicity of pollutants emitted by the exhaust gases, with an increase in total protein, proline and stimulation of detoxification enzyme (catalase).

Keywords: air pollution, toxicity, epidemiology, biochemistry

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664 Critical Role of Lipid Rafts in Influenza a Virus Binding to Host Cell

Authors: Dileep Kumar Verma, Sunil Kumar Lal

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Influenza still remains one of the most challenging diseases posing significant threat to public health causing seasonal epidemics and pandemics. Influenza A Virus (IAV) surface protein hemagglutinin is known to play an important role in viral attachment to the host sialic acid receptors and concentrate in lipid rafts for efficient viral fusion. Selective nature of Influenza A virus to utilize rafts micro-domain for efficient virus assembly and budding has been explored in depth. However, the detailed mechanism of IAV binding to host cell membrane and entry into the host remains elusive. In the present study we investigated the role of lipid rafts in early life cycle events of IAV. Role of host lipid rafts was studied using raft disruption method by extraction of cholesterol by Methyl-β-Cyclodextrin. Using GM1, a well-known lipid raft marker, we were able to observe co-localization of IAV on lipid rafts on the host cell membrane. This experiment suggests a direct involvement of lipid rafts in the initiation of the IAV life cycle. Upon disruption of lipid rafts by Methyl-b-cyclodextrin, we observed a significant reduction in IAV binding on the host cell surface indicating a significant decrease in virus attachment to coherent membrane rafts. Our results provide proof that host lipid rafts and their constituents play an important role in the adsorption of IAV. This study opens a new avenues in IAV virus-host interactions to combat infection at a very early steps of the viral lifecycle.

Keywords: lipid raft, adsorption, cholesterol, methyl-β-cyclodextrin, GM1

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663 Processing Methods for Increasing the Yield, Nutritional Value and Stability of Coconut Milk

Authors: Archana G. Lamdande, Shyam R. Garud, K. S. M. S. Raghavarao

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Coconut has two edible parts, that is, a white kernel (solid endosperm) and coconut water (liquid endosperm). The white kernel is generally used in fresh or dried form for culinary purposes. Coconut testa, is the brown skin, covering the coconut kernel. It is removed by paring of wet coconut and obtained as a by-product in coconut processing industries during the production of products such as desiccated coconut, coconut milk, whole coconut milk powder and virgin coconut oil. At present, it is used as animal feed component after drying and recovering the residual oil (by expelling). Experiments were carried out on expelling of coconut milk for shredded coconut with and without testa removal, in order to explore the possibility of increasing the milk yield and value addition in terms of increased polyphenol content. The color characteristics of coconut milk obtained from the grating without removal of testa were observed to be L* 82.79, a* 0.0125, b* 6.245, while that obtained from grating with removal of testa were L* 83.24, a* -0.7925, b* 3.1. A significant increase was observed in total phenol content of coconut milk obtained from the grating with testa (833.8 µl/ml) when compared to that from without testa (521.3 µl/ml). However, significant difference was not observed in protein content of coconut milk obtained from the grating with and without testa (4.9 and 5.0% w/w, respectively). Coconut milk obtained from grating without removal of testa showed higher milk yield (62% w/w) when compared to that obtained from grating with removal of testa (60% w/w). The fat content in coconut milk was observed to be 32% (w/w), and it is unstable due to such a high fat content. Therefore, several experiments were carried out for examining its stability by adjusting the fat content at different levels (32, 28, 24, and 20% w/w). It was found that the coconut milk was more stable with a fat content of 24 % (w/w). Homogenization and ultrasonication and their combinations were used for exploring the possibility of increasing the stability of coconut milk. The microscopic study was carried out for analyzing the size of fat globules and the degree of their uniform distribution.

Keywords: coconut milk, homogenization, stability, testa, ultrasonication

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662 Network Pharmacological Evaluation of Holy Basil Bioactive Phytochemicals for Identifying Novel Potential Inhibitors Against Neurodegenerative Disorder

Authors: Bhuvanesh Baniya

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Alzheimer disease is illnesses that are responsible for neuronal cell death and resulting in lifelong cognitive problems. Due to their unclear mechanism, there are no effective drugs available for the treatment. For a long time, herbal drugs have been used as a role model in the field of the drug discovery process. Holy basil in the Indian medicinal system (Ayurveda) is used for several neuronal disorders like insomnia and memory loss for decades. This study aims to identify active components of holy basil as potential inhibitors for the treatment of Alzheimer disease. To fulfill this objective, the Network pharmacology approach, gene ontology, pharmacokinetics analysis, molecular docking, and molecular dynamics simulation (MDS) studies were performed. A total of 7 active components in holy basil, 12 predicted neurodegenerative targets of holy basil, and 8063 Alzheimer-related targets were identified from different databases. The network analysis showed that the top ten targets APP, EGFR, MAPK1, ESR1, HSPA4, PRKCD, MAPK3, ABL1, JUN, and GSK3B were found as significant target related to Alzheimer disease. On the basis of gene ontology and topology analysis results, APP was found as a significant target related to Alzheimer’s disease pathways. Further, the molecular docking results to found that various compounds showed the best binding affinities. Further, MDS top results suggested could be used as potential inhibitors against APP protein and could be useful for the treatment of Alzheimer’s disease.

Keywords: holy basil, network pharmacology, neurodegeneration, active phytochemicals, molecular docking and simulation

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661 Growth Performance and Nutrient Digestibility of Cirrhinus mrigala Fingerlings Fed on Sunflower Meal Based Diet Supplemented with Phytase

Authors: Syed Makhdoom Hussain, Muhammad Afzal, Farhat Jabeen, Arshad Javid, Tasneem Hameed

Abstract:

A feeding trial was conducted with Cirrhinus mrigala fingerlings to study the effects of microbial phytase with graded levels (0, 500, 1000, 1500, and 2000 FTUkg-1) by sunflower meal based diet on growth performance and nutrient digestibility. The chromic oxide was added as an indigestible marker in the diets. Three replicate groups of 15 fish (Average wt 5.98 g fish-1) were fed once a day and feces were collected twice daily. The results of present study showed improved growth and feed performance of Cirrhinus mrigala fingerlings in response to phytase supplementation. Maximum growth performance was obtained by the fish fed on test diet-III having 1000 FTU kg-1 phytase level. Similarly, nutrient digestibility was also significantly increased (p<0.05) by phytase supplementation. Digestibility coefficients for sunflower meal based diet increased 15.76%, 17.70%, and 12.70% for crude protein, crude fat and apparent gross energy as compared to the reference diet, respectively at 1000 FTU kg-1 level. Again, maximum response of nutrient digestibility was recorded at the phytase level of 1000 FTU kg-1 diet. It was concluded that the phytase supplementation to sunflower meal based diet at 1000 FTU kg-1 level is optimum to release adequate chelated nutrients for maximum growth performance of C. mrigala fingerlings. Our results also suggested that phytase supplementation to sunflower meal based diet can help in the development of sustainable aquaculture by reducing the feed cost and nutrient discharge through feces in the aquatic ecosystem.

Keywords: sunflower meal, Cirrhinus mrigala, growth, nutrient digestibility, phytase

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660 Subclinical Renal Damage Induced by High-Fat Diet in Young Rats

Authors: Larissa M. Vargas, Julia M. Sacchi, Renata O. Pereira, Lucas S. Asano, Iara C. Araújo, Patricia Fiorino, Vera Farah

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The aim of this study was to evaluate the occurrence of subclinical organ injuries induced by high-fat diet. Male wistar rats (n=5/group) were divided in control diet group (CD), commercial rat chow, and hyperlipidic diet (30% lipids) group (HD) administrated during 8 weeks, starting after weaning. All the procedures followed the rules of the Committee of Research and Ethics of the Mackenzie University (CEUA Nº 077/03/2011). At the end of protocol the animals were euthanized by anesthesia overload and the left kidney was removed. Intrarenal lipid deposition was evaluated by histological analyses with oilred. Kidney slices were stained with picrosirius red to evaluate the area of the Bowman's capsule (AB) and space (SB), and glomerular tuft area (GT). The renal expression of sterol regulatory element–binding protein (SREBP-2) was performed by Western Blotting. Creatinine concentration (serum and urine) and lipid profile were determined by colorimetric kit (Labtest). At the end of the protocol there was no differences in body weight between the groups, however the HD showed a marked increase in lipid deposits, glomeruli and tubules, and biochemical analysis for cholesterol and triglycerides. Moreover, in the kidney, the high-fat diet induced a reduction in the AB (13%), GT (18%) and SB (17%) associated with a reduction in glomerular filtration rate (creatinine clearance). The renal SRBP2 expression was increased in HD group. These data suggests that consumption of high-fat diet starting in childhood is associated with subclinical renal damage and function.

Keywords: high-fat diet, kidney, intrarenal lipid deposition, SRBP2

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659 Impact of Environmental Changes on Blood Parameters in the Pelophylax ridibundus

Authors: Murat Tosunoglu, Cigdem Gul, Nurcihan Hacioglu, Nurdan Tepeova

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Amphibian and Reptilian species are influenced by pollution and habitat destruction. Blood parameters of Amphibia species were particularly affected by the negative environmental conditions. Studied frog samples 36 clinically normal Pelophylax ridibundus individuals were captured along the Biga Stream between April–June 2014. When comparing our findings with the Turkish legislation (Water pollution control regulation), the 1. Locality of the Biga stream in terms of total coliform classified as "high quality water" (Coliform: 866.66 MPN/100 mL), while the 2. locality was a "contaminated water" (Coliform: 53266.66 MPN/100 mL). Blood samples of the live specimens were obtained in the laboratory within one day of their capture. The blood samples were taken from the etherized frogs by means of ventriculus punctures, via heparinized hematocrit capillaries. Hematological and biochemical analyses based on high quality water and contaminated water, respectively, are as follows: Red blood cell count (444210.52-426846.15 per cubic millimeter of blood), white blood cell count (4215.78-4684.61 per cubic millimeter of blood), hematocrit value (29.25-29.43 %), hemoglobin concentration (7.76-7.22 g/dl), mean corpuscular volume (637.64-719.99 fl), mean corpuscular hemoglobin (184.78-174.75 pg), mean corpuscular hemoglobin concentration (29.44-24.82 %), glucose (103.74-124.13 mg/dl), urea (87.68-81.72 mg/L), cholesterol (148.20-197.39 mg/dl), creatinine (0.29-0.28 mg/dl), uric acid (10.26-7.55 mg/L), albumin (1.13-1.39 g/dl), calcium (11.45-9.70 mg/dl), triglyceride (135.23-155.85 mg/dl), total protein (4.26-3.73 g/dl), phosphorus (6.83-17.86 mg/dl), and magnesium (0.95-1.06 mg/dl). The some hematological parameters in P. ridibundus specimens are given for the first time in this study. No water quality dependent variation was observed in clinic hematology parameters measured.

Keywords: Pelophylax ridibundus, hematological parameters, biochemistry, freshwater quality

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658 Distributing Complementary Food Supplement - Yingyangbao Reducing the Anemia in Young Children in a County of Sichuan Province after Wenchuan Earthquake

Authors: Lijuan Wang, Junsheng Huo, Jing Sun, Wenxian Li, Jian Huang, Lin Ling, Yiping Zhou, Chengyu Huang, Jifang Hu

Abstract:

Backgrounds and Objective: This study aimed to evaluate the impact of highly nutrient-dense complementary food supplement-Yingyangbao, at the time of 3 months after Wenchuan earthquake, on the anemia of young children in a county in Sichuan province. Methods: The young children aged 6-23 months in the county were fed one sachet Yingyangbao per day. Yingyangbao were distributed for 15 months for free. The children entering 6 months age would be included. The length, weight and hemoglobin of the children aged 6-29 months were assessed at baseline (n=257) and Yingyangbao intervention for 6 (n=218) and 15 months (n=253) by cluster sampling. Growth status has not been described in the paper. The analysis was conducted based on 6-11, 12-17, 18-23 and 24-29 months. Results: It showed that the hemoglobin concentration in each group among the 4 groups increased by 4.9, 6.4, 8.0, 9.5 g/L after 6 months and 12.7, 11.4, 16.7, 15.7 g/L after 15 months compared to the baseline, respectively. The total anemia prevalence in each group was significantly lower after 6 and 15 months than the baseline (P<0.001), except the 6-11 months group after 6 months because of fewer Yingyangbao consumption. Total moderate anemia rate decreased from 18.3% to 5.5% after 6 months, and kept decreasing to 0.8% after another 9 months. The hemoglobin concentration was significantly correlated with the amount of Yingyangbao consumption(P<0.001) The anemia rate was significantly different based on the Yingyangbao compliance (P<0.001). Conclusion: It was concluded that Yingyangbao which contains quality protein, vitamins and micronutrients intervened 15 months could be effective for the improvement of anemia of young children. The study provides the support that the application of the complementary food supplements to reduce the anemia of young children in the emergency of natural disaster.

Keywords: young children, anemia, nutrition intervention, complementary food supplements, Yingyangbao

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657 Effects of Ethylene Scavengering Packaging on the Shelf Life of Edible Mushroom

Authors: Majid Javanmard

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Edible mushrooms are those agricultural products which contain high quantity of protein and can have special role in human diet. So search for methods to increase their shelf life is important. One of these strategies can be use of active packaging for absorb the ethylene which has been studied in present study. In this study, initially, production of impregnating zeolite with potassium permanganate has been studied with zeolite clinoptiolite available in iran. After that, these ethylene scavengers were placed in the package of edible mushrooms and then transferred to the refrigerator with temperature 4c for a period of 20 days. Each 5 days, several experiments accomplished on edible mushrooms such as weight loss, moisture content, color, texture, bacterial experiments and sensory evaluation. After production of impregnating zeolite with potassium permanganate (with a concentration of %2.5, %5, %7.5, %10 and %12.5) by zeolite type clinoptiolite (with mesh 35 and 60), samples have been analyzed with gas chromatography and titration with sodium oxalate. The results showed that zeolite by concentration of %5, %7.5 and %10 potassium permanganate and mesh 60 have a higher efficiency. Results from the experiments on edible mushrooms proved that impregnated zeolite with potassium permanganate have a meaningful influence in prevent the weight loss, decrease of moisture content and L-value, increase of a-value and overall color change (ΔE) and decrease of firmness texture of mushrooms. In addition, these absorbents can influence on decrease microbial load (mesophilic bacteria) rather than control. Generally, concluded that the impregnated zeolite with 10% permanganate potassium has a high efficiency on increase the shelf life of fresh edible mushrooms.

Keywords: active packaging, ethylene scavenger, zeolite clinoptiolite, permanganate potassium, shelf life

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656 Effects of Microbial Biofertilization on Nodulation, Nitrogen Fixation, and Yield of Lablab purpureus

Authors: Benselama Amel, Ounane S. Mohamed, Bekki Abdelkader

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A collection of 20 isolates from fresh Nodules of the legume plant Lablab purpureus was isolated. These isolates have been authenticated by seedling inoculation grown in jars containing sand. The results obtained after two months of culture have revealed that the 20 isolates (100% of the isolates) are able to nodulate their host plants. The results obtained were analyzed statistically by ANOVA using the software statistica and had shown that the effect of the inoculation has significantly improved all the growth parameters (the height of the plant and the dry weight of the aerial parts and roots, and the number of nodules). We have evaluated the tolerance of all strains of the collection to the major stress factors as the salinity, pH and extreme temperature. The osmotolerance reached a concentration up to 1710mm of NaCl. The strains were also able to grow on a wide range of pH, ranging from 4.5 to 9.5, and temperature, between 4°C and 40°C. Also, we tested the effect of the acidity, aluminum and ferric deficit on the Lablab-rhizobia symbiosis. Lablab purpureus has not been affected by the presence of high concentrations of aluminum. On the other hand, iron deficiency has caused a net decrease in the dry biomass of the aerial part. The results of all the phenotypic characters have been treated by the statistical Minitab software, the numerical analysis had shown that these bacterial strains are divided into two distinct groups at a level of similarity of 86 %. The SDS-PAGE was carried out to determine the profile of the total protein of the strains. The coefficients of similarity of polypeptide bands between the isolates and strains reference (Bradyrhizobium, Mesorizobium sp.) confirm that our strain belongs to the groups of rhizobia.

Keywords: SDS-PAGE, rhizobia, symbiosis, phenotypic characterization, Lablab purpureus

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655 Humoral and Cytokine Responses to Major Human Cytomegalovirus Antigens in Mouse Model

Authors: Sahar Essa, Hussain A. Safar, Raj Raghupathy

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Human cytomegalovirus (CMV) continues to be a source of severe complications in immunologically immature and immunocompromised hosts. Effective CMV vaccines that help diminish CMV disease in transplant patients and avoid congenital infection are of great importance. Though the exact roles of defense mechanisms are unidentified, viral-specific antibodies and cytokine responses are known to be involved in controlling CMV infections. CMV envelope glycoprotein B (UL55/gB), matrix proteins (UL83/pp65, UL99/pp28, UL32/pp150), and assembly protein UL80a/pp38 are known to be targets of antiviral immune responses. We immunized mice intraperitoneally with these five CMV-related proteins (commercial) for their ability to induce specific antibody responses (in-house immunoassay) and cytokine production (commercial assay) in a mouse model. We observed a significant CMV-antigen-specific antibody response to pp38 and pp65 (E/C ˃2.0, p˂0.001). Mice immunized with pp38 had significantly higher concentrations of GM-CSF, IFN-α, IL-2 IL-4, IL-5, and IL-17A (p˂0.05). Mice immunized with pp65 showed significantly higher concentrations of GM-CSF, IFN-γ, IL-2 IL-4, IL-10, IL-12, IL-17A, and TNF-α. Th1 to Th2 cytokines ratios revealed a Th1 cytokine bias in mice immunized with pp38, pp65, pp150, and gB. We suggest that stimulation with multiple CMV-related proteins, which include pp38, pp65, and gB antigens, will allow both humoral and cellular immune responses to be efficiently activated, thus serving as appropriate CMV antigens for future vaccines.

Keywords: cytomegalovirus, UL99/pp28, UL80a/pp38, UL83/pp65, UL32/pp150, UL55/gB, CMV-antigen-specific antibody, CMV antigen-specific cytokine responses

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