Search results for: PTP1B in-vitro anti-diabetic assay female albino mice
Commenced in January 2007
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Paper Count: 4272

Search results for: PTP1B in-vitro anti-diabetic assay female albino mice

3042 Prognostic Value in Meningioma Patients’: A Clinical-Histopathological Study

Authors: Ilham Akbar Rahman, Aflah Dhea Bariz Yasta, Iin Fadhilah Utami Tamasse, Devina Juanita

Abstract:

Meningioma is adult brain tumors originating from the meninges covering the brain and spinal cord. The females have approximately twice higher 2:1 than male in the incidence of meningioma. This study aimed to analyze the histopathological grading and clinical aspect in predicting the prognosis of meningioma patients. An observational study with cross sectional design was used on 53 meningioma patients treated at Dr. Wahidin Sudirohusodo hospital in 2016. The data then were analyzed using SPSS 20.0. Of 53 patients, mostly 41 (77,4%) were female and 12 (22,6%) were male. The distribution of histopathology patients showed the meningothelial meningioma of 18 (43,9%) as the most type found. Fibroplastic meningioma were 8 (19,5%), while atypical meningioma and psammomatous meningioma were 6 (14,6%) each. The rest were malignant meningioma and angiomatous meningioma which found in respectively 2 (4,9%) and 1 (2,4%). Our result found significant finding that mostly male were fibroblastic meningioma (50%), however meningothelial meningioma were found in the majority of female (54,8%) and also seizure comprised only in higher grade meningioma. On the outcome of meningioma patient treated operatively, histopathological grade remained insignificant (p > 0,05). This study can be used as prognostic value of meningioma patients based on gender, histopathological grade, and clinical manifestation. Overall, the outcome of the meningioma’s patients is good and promising as long as it is well managed.

Keywords: meningioma, prognostic value, histopathological grading, clinical manifestation

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3041 Prevalence of Clostridium perfringens β2-Toxin in Type a Isolates of Sheep and Goats

Authors: Mudassar Mohiuddin, Zahid Iqbal

Abstract:

Introduction: Clostridium perfringens is an important pathogen responsible for causing enteric diseases in both human and animals. The bacteria produce several toxins. These toxins play vital role in the pathogenesis of various fatal enteric diseases and are classified into five types, on the basis of the differential production of Alpha, Beta, Epsilon and Iota toxins. In addition to the so-called major toxins, there are other toxins like beta2 toxin, produced by some strains of C. perfringens which may play a role in the pathogenesis of disease. Aim of the study: In this study a multiplex PCR assay was developed and used for detection of cpb2 gene to identify the Beta2 harboring isolates among different types of C. perfringens. Objectives: The primary objective of this study was to identify the prevalence of β2-toxin gene in local isolates of Clostridium perfringens. Methodology: This was an experimental study. Random sampling technique was used. A total of 97 sheep and goats were included in this study. All were Pakistani local breeds. The samples were collected during the period from Sep, 2014 to Mar, 2015 from selected districts of Punjab province (Pakistan). Faecal samples were cultured in cooked meat media. The identification of Clostridium perfringens was made on the basis of biochemical tests. Multiplex PCR was performed to identify the toxin genes. Results: A total of 43 C. perfringens isolates were genotyped using multiplex PCR assay. The gene encoding C. perfringens β2-toxin (cpb2) was present in more than 50% of the isolates genotyped. However, the prevalence of this gene varied between sheep and goat isolates. Conclusion: The present study suggests the high occurrence of C. perfringens b2-toxin (cpb2) in the local isolates of Pakistan. As β2-toxin is present in both healthy and diseased animals, so further studies are suggested to establish the role of β2-toxin in pathogenesis of the clostridial enteric diseases.

Keywords: beta 2 toxin gene, clostridium perfringens, enteric diseases, goats, multiplex PCR, sheep

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3040 Neuro-Epigenetic Changes on Diabetes Induced-Synaptic Fidelity in Brain

Authors: Valencia Fernandes, Dharmendra Kumar Khatri, Shashi Bala Singh

Abstract:

Background and Aim: Epigenetics are the inaudible signatures of several pathological processes in the brain. This study understands the influence of DNA methylation, a major epigenetic modification, in the prefrontal cortex and hippocampus of the diabetic brain and its notable effect on the cellular chaperones and synaptic proteins. Method: Chronic high fat diet and STZ-induced diabetic mice were studied for cognitive dysfunction, and global DNA methylation, as well as DNA methyltransferase (DNMT) activity, were assessed. Further, the cellular chaperones and synaptic proteins were examined using DNMT inhibitor, 5-aza-2′-deoxycytidine (5-aza-dC)-via intracerebroventricular injection. Moreover, % methylation of these synaptic proteins were also studied so as to correlate its epigenetic involvement. Computationally, its interaction with the DNMT enzyme were also studied using bioinformatic tools. Histological studies for morphological alterations and neuronal degeneration were also studied. Neurogenesis, a characteristic marker for new learning and memory formation, was also assessed via the BrdU staining. Finally, the most important behavioral studies, including the Morris water maze, Y maze, passive avoidance, and Novel object recognition test, were performed to study its cognitive functions. Results: Altered global DNA methylation and increased levels of DNMTs within the nucleus were confirmed in the cortex and hippocampus of the diseased mice, suggesting hypermethylation at a genetic level. Treatment with AzadC, a global DNA demethylating agent, ameliorated the protein and gene expression of the cellular chaperones and synaptic fidelity. Furthermore, the methylation analysis profile showed hypermethylation of the hsf1 protein, a master regulator for chaperones and thus, confirmed the epigenetic involvement in the diseased brain. Morphological improvements and decreased neurodegeneration, along with enhanced neurogenesis in the treatment group, suggest that epigenetic modulations do participate in learning and memory. This is supported by the improved behavioral test battery seen in the treatment group. Conclusion: DNA methylation could possibly accord in dysregulating the memory-associated proteins at chronic stages in type 2 diabetes. This could suggest a substantial contribution to the underlying pathophysiology of several metabolic syndromes like insulin resistance, obesity and also participate in transitioning this damage centrally, such as cognitive dysfunction.

Keywords: epigenetics, cognition, chaperones, DNA methylation

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3039 A Comparison of Sulfur Mustard Cytotoxic Effects on the Two Human Lung Origin Cell Lines

Authors: P. Jost, L. Muckova, M. Matula, J. Pejchal, D. Jun, R. Stetina

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Sulfur mustard (bis(2-chlorethyl) sulfide) is highly toxic, chemical warfare agent that has been used in the past in several armed conflicts. Except for the skin, respiratory tract is one of the important routes of exposure. The elucidation and understanding of the mechanism of toxicity of SM have been effort intensive research. The multiple targets character of SM caused cellular damage resulted in activation of many different mechanisms which contribute to cellular response and participate in the final cytopathology effect. In our present work, we compared time-dependent changes in sulfur mustard exposed adult human lung fibroblasts NHLF and lung epithelial alveolar cell line A-549. Cell viability (MTT assay, Calcein-AM assay, and xCELLigence - real-time cell analysis), apoptosis (flow cytometry), mitochondrial membrane potential (Δψm, flow cytometry), reactive oxygen species induction (DC and cell cycle distribution (flow cytometry) were studied. We observed significantly decreased mitochondrial membrane potential and subsequent induction of apoptosis correlating with decreased cellular viability in the sulfur mustard exposed cells. In low concentrations, sulfur mustard-induced S-phase cell cycle arrest, on the other hand, high concentrations, cell cycle phase distribution of sulfur mustard exposed cells resembled cell cycle phase distribution of control group, which implies nonspecific cell cycle inhibition. Epithelial cells A-549 was found as more sensible to sulfur mustard toxicity. Acknowledgements: This work was supported by a long-term organization development plan Medical Aspects of Weapons of Mass Destruction of the Faculty of Military Health Sciences, University of Defence.

Keywords: apoptosis, cell cycle, cytotoxicity, sulfur mustard

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3038 Daye™ Tampon as a Tool for Vaginal Sample Collection Towards the Detection of Genital Infections

Authors: Valentina Milanova, Kalina Mihaylova, Iva Lazarova

Abstract:

The mechanisms by which female genital infections are detected are varied and include clinician-collected high vaginal swabs, clinician-collected endocervical swabs, patient-collected vaginal swabs, and first-pass urine samples. Vaginal health screening has chronically low rates of uptake. This highlights the unmet need for a screening tool with comparable diagnostic accuracy which is familiar, convenient and easy to use for people. The Daye™ medical grade tampon offers an alternative to traditional sampling methods with the potential of increasing screening uptake among people previously too embarrassed or busy to attend gynecological appointments. In this white paper, the results of stability studies and a comparative clinical trial are discussed to assess the suitability of the device for the collection of vaginal samples for various clinical assessments. The tampon has demonstrated good sample stability and comparable sample quality compared to a self-collected vaginal swab and a clinician-collected cervical swab.

Keywords: vaginal microbiome, vaginal infections, gynaecological infections, female health, menstrual tampons, in vitro diagnostics

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3037 Gynocentrism and Self-Orientalization: A Visual Trend in Chinese Fashion Photography

Authors: Zhen Sun

Abstract:

The study adopts the method of visual social semiotics to analyze a sample of fashion photos that were recently published in Chinese fashion magazines that target towards both male and female readers. It identifies a new visual trend in fashion photography, which is characterized by two features. First, the photos represent young, confident, and stylish female models with lower-class sloppy old men. The visual inharmony between the sexually desirable women and the aged men has suggested an impossibly accomplished sexuality and eroticism. Though the women are still under the male gaze, they are depicted as unreachable objects of voyeurism other than sexual objects subordinated to men. Second, the represented people are usually put in the backdrop of tasteless or vulgar Chinese town life, which is congruent with the images of men but makes the modern city girls out of place. The photographers intentionally contrast the images of women with that of men and with the background, which implies an imaginary binary division of modern Orientalism and the photographers’ self-orientalization strategy. Under the theoretical umbrella of neoliberal postfeminism, this study defines a new kind of gynocentric stereotype in Chinese fashion photography, which challenges the previous observations on gender portrayals in fashion magazines.

Keywords: fashion photography, gynocentrism, neoliberal postfeminism, self-orientalization

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3036 Characterizing and Developing the Clinical Grade Microbiome Assay with a Robust Bioinformatics Pipeline for Supporting Precision Medicine Driven Clinical Development

Authors: Danyi Wang, Andrew Schriefer, Dennis O'Rourke, Brajendra Kumar, Yang Liu, Fei Zhong, Juergen Scheuenpflug, Zheng Feng

Abstract:

Purpose: It has been recognized that the microbiome plays critical roles in disease pathogenesis, including cancer, autoimmune disease, and multiple sclerosis. To develop a clinical-grade assay for exploring microbiome-derived clinical biomarkers across disease areas, a two-phase approach is implemented. 1) Identification of the optimal sample preparation reagents using pre-mixed bacteria and healthy donor stool samples coupled with proprietary Sigma-Aldrich® bioinformatics solution. 2) Exploratory analysis of patient samples for enabling precision medicine. Study Procedure: In phase 1 study, we first compared the 16S sequencing results of two ATCC® microbiome standards (MSA 2002 and MSA 2003) across five different extraction kits (Kit A, B, C, D & E). Both microbiome standards samples were extracted in triplicate across all extraction kits. Following isolation, DNA quantity was determined by Qubit assay. DNA quality was assessed to determine purity and to confirm extracted DNA is of high molecular weight. Bacterial 16S ribosomal ribonucleic acid (rRNA) amplicons were generated via amplification of the V3/V4 hypervariable region of the 16S rRNA. Sequencing was performed using a 2x300 bp paired-end configuration on the Illumina MiSeq. Fastq files were analyzed using the Sigma-Aldrich® Microbiome Platform. The Microbiome Platform is a cloud-based service that offers best-in-class 16S-seq and WGS analysis pipelines and databases. The Platform and its methods have been extensively benchmarked using microbiome standards generated internally by MilliporeSigma and other external providers. Data Summary: The DNA yield using the extraction kit D and E is below the limit of detection (100 pg/µl) of Qubit assay as both extraction kits are intended for samples with low bacterial counts. The pre-mixed bacterial pellets at high concentrations with an input of 2 x106 cells for MSA-2002 and 1 x106 cells from MSA-2003 were not compatible with the kits. Among the remaining 3 extraction kits, kit A produced the greatest yield whereas kit B provided the least yield (Kit-A/MSA-2002: 174.25 ± 34.98; Kit-A/MSA-2003: 179.89 ± 30.18; Kit-B/MSA-2002: 27.86 ± 9.35; Kit-B/MSA-2003: 23.14 ± 6.39; Kit-C/MSA-2002: 55.19 ± 10.18; Kit-C/MSA-2003: 35.80 ± 11.41 (Mean ± SD)). Also, kit A produced the greatest yield, whereas kit B provided the least yield. The PCoA 3D visualization of the Weighted Unifrac beta diversity shows that kits A and C cluster closely together while kit B appears as an outlier. The kit A sequencing samples cluster more closely together than both the other kits. The taxonomic profiles of kit B have lower recall when compared to the known mixture profiles indicating that kit B was inefficient at detecting some of the bacteria. Conclusion: Our data demonstrated that the DNA extraction method impacts DNA concentration, purity, and microbial communities detected by next-generation sequencing analysis. Further microbiome analysis performance comparison of using healthy stool samples is underway; also, colorectal cancer patients' samples will be acquired for further explore the clinical utilities. Collectively, our comprehensive qualification approach, including the evaluation of optimal DNA extraction conditions, the inclusion of positive controls, and the implementation of a robust qualified bioinformatics pipeline, assures accurate characterization of the microbiota in a complex matrix for deciphering the deep biology and enabling precision medicine.

Keywords: 16S rRNA sequencing, analytical validation, bioinformatics pipeline, metagenomics

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3035 Immunostimulatory Response of Supplement Feed in Fish against Aeromonas hydrophila

Authors: Shikha Rani, Neeta Sehgal, Vipin Kumar Verma, Om Prakash

Abstract:

Introduction: Fish is an important protein source for humans and has great economic value. Fish cultures are affected due to various anthropogenic activities that lead to bacterial and viral infections. Aeromonas hydrophila is a fish pathogenic bacterium that causes several aquaculture outbreaks throughout the world and leads to huge mortalities. In this study, plants of no commercial value were used to investigate their immunostimulatory, antioxidant, anti-inflammatory, anti-bacterial, and disease resistance potential in fish against Aeromonas hydrophila, through fish feed fortification. Methods: The plant was dried at room temperature in the shade, dissolved in methanol, and analysed for biological compounds through GC-MS/MS. DPPH, FRAP, Phenolic, and flavonoids were estimated following standardized protocols. In silico molecular docking was also performed to validate its broad-spectrum activities based on binding affinity with specific proteins. Fish were divided into four groups (n=6; total 30 in a group): Group 1, non-challenged fish (fed on a non-supplemented diet); Group 2, fish challenged with bacteria (fed on a non-supplemented diet); Group 3 and 4, fish challenged with bacteria (A. hydrophila) and fed on plant supplemented feed at 2.5% and 5%. Blood was collected from the fish on 0, 7th, 14th, 21st, and 28th days. Serum was separated for glutamic-oxaloacetic transaminase (SGOT), serum glutamic pyruvic transaminase (SGPT), alkaline phosphatase assay (ALP), lysozyme activity assay, superoxide dismutase assay (SOD), lipid peroxidation assay (LPO) and molecular parameters (including cytokine levels) were estimated through ELISA. The phagocytic activity of macrophages from the spleen and head kidney, along with quantitative analysis of immune-related genes, were analysed in different tissue samples. The digestive enzymes (Pepsin, Trypsin, and Chymotrypsin) were also measured to evaluate the effect of plant-supplemented feed on freshwater fish. Results and Discussion: GC-MS/MS analysis of a methanolic extract of plant validated the presence of key compounds having antioxidant, anti-inflammatory, anti-bacterial, anti-inflammatory, and immunomodulatory activities along with disease resistance properties. From biochemical investigations like ABTS, DPPH, and FRAP, the amount of total flavonoids, phenols, and promising binding affinities towards different proteins in molecular docking analysis helped us to realize the potential of this plant that can be used for investigation in the supplemented feed of fish. Measurement liver function tests, ALPs, oxidation-antioxidant enzyme concentrations, and immunoglobulin concentrations in the experimental groups (3 and 4) showed significant improvement as compared to the positive control group. The histopathological evaluation of the liver, spleen, and head kidney supports the biochemical findings. The isolated macrophages from the group fed on supplemented feed showed a higher percentage of phagocytosis and a phagocytic index, indicating an enhanced cell-mediated immune response. Significant improvements in digestive enzymes were also observed in fish fed on supplemented feed, even after weekly challenges with bacteria. Hence, the plant-fortified feed can be recommended as a regular feed to enhance fish immunity and disease resistance against the Aeromonas hydrophila infection after confirmation from the field trial.

Keywords: immunostimulation, antipathogen, plant fortified feed, macrophages, GC-MS/MS, in silico molecular docking

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3034 Gross and Clinical Anatomy of the Skull of Adult Chinkara, Gazella bennettii

Authors: Salahud Din, Saima Masood, Hafsa Zaneb, Habib Ur Rehman, Saima Ashraf, Imad Khan, Muqader Shah

Abstract:

The objective of this study was (1) to study gross morphological, osteometric and clinical important landmarks in the skull of adult Chinkara to obtain baseline data and (2) to study sexual dimorphism in male and female adult Chinkara through osteometry. For this purpose, after performing postmortem examination, the carcass of adult Chinkara of known sex and age was buried in the locality of the Manglot Wildlife Park and Ungulate Breeding Centre, Nizampur, Pakistan; after a specific period of time, the bones were unearthed. Gross morphological features and various osteometric parameters of the skull were studied in the University of Veterinary and Animal Sciences, Lahore, Pakistan. The shape of the Chinkara skull was elongated and had thirty-two bones. The skull was comprised of the cranial and the facial part. The facial region of the skull was formed by maxilla, incisive, palatine, vomar, pterygoid, frontal, parietal, nasal, incisive, turbinates, mandible and hyoid apparatus. The bony region of the cranium of Chinkara was comprised of occipital, ethmoid, sphenoid, interparietal, parietal, temporal, and frontal bone. The foramina identified in the facial region of the skull of Chinkara were infraorbital, supraorbital foramen, lacrimal, sphenopalatine, maxillary and caudal palatine foramina. The foramina of the cranium of the skull of the Chinkara were the internal acoustic meatus, external acoustic meatus, hypoglossal canal, transverse canal, sphenorbital fissure, carotid canal, foramen magnum, stylomastoid foramen, foramen rotundum, foramen ovale and jugular foramen, and the rostral and the caudal foramina that formed the pterygoid canal. The measured craniometric parameters did not show statistically significant differences (p > 0.05) between male and female adult Chinkara except Palatine bone, OI, DO, IOCDE, OCT, ICW, IPCW, and PCPL were significantly higher (p > 0.05) in male than female Chinkara and mean values of the mandibular parameters except b and h were significantly (p < 0.5) higher in male Chinkara than female Chinkara. Sexual dimorphism exists in some of the orbital and foramen magnum parameters, while high levels of sexual dimorphism identified in mandible. In conclusion, morphocraniometric studies of Chinkara skull made it possible to identify species-specific skull and use clinical measurements during practical application.

Keywords: Chinkara, skull, morphology, morphometrics, sexual dimorphism

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3033 Let-7 Mirnas Regulate Inflammatory Cytokine Production in Bovine Endometrial Cells after Lipopolysaccharide Challenge by Targeting TNFα

Authors: S. Ibrahim, D. Salilew-Wondim, M. Hoelker, C. Looft, E. Tholen, C. Grosse-Brinkhaus, K. Schellander, C. Neuhoff, D. Tesfaye

Abstract:

Bovine endometrial cells appear to have a key role in innate immune defense of the female genital tract. A better understanding of molecular changes in microRNAs (miRNAs) and their target genes expression may identify reliable prognostic indicators for cows that will resolve inflammation and resume cyclicity. In the current study, we hypothesized that let-7 miRNAs family has a primary role in the innate immune defence of the endometrium tissue against bacterial infection, which is partly achieved via regulating mRNA stability of pro-inflammatory cytokines at the post-transcriptional level. Therefore, we conducted two experiments. In the first experiment, primary bovine endometrial cells were challenged with clinical (3.0 μg/ml) and sub-clinical (0.5 μg/ml) doses of lipopolysaccharide (LPS) for 24h. In the 2nd experiment, we have investigated the potential role of let-7 miRNAs (let-7a and let-7f) using gain and loss of function approaches. Additionally, tumor necrosis factor alpha (TNFα), transforming growth factor beta 1 induced transcript 1 (TGFB1I1) and serum deprivation response (SDPR) genes were validated using reporter assay. Here we addressed for the first time that let-7 miRNAs have a precise role in bovine endometrium, where LPS dysregulated let-7 miRNAs family expression was associated with an increased pro-inflammatory cytokine level by directly/indirectly targeting the TNFα, interleukin 6 (IL6), nuclear factor kappa-light-chain enhancer of activated B cells (NFκB), TGFβ1I1 and SDPR genes. To our knowledge, this is the first study showing that TNFα, TGFβ1I1 and SDPR were identified and validated as novel let-7 miRNAs targets and could have a distinct role in inflammatory immune response of LPS challenged bovine endometrial cells. Our data represent a new finding by which uterine homeostasis is maintained through functional regulation of let-7a by down-regulation of pro-inflammatory cytokines expression (TNFα and IL6) at the mRNA and protein levels. These findings suggest that LPS serves as a negative regulator of let-7 miRNAs expression and provides a mechanism for the persistent pro-inflammatory phenotype, which is a hallmark of bovine subclinical endometritis.

Keywords: bovine endometrial cells, let-7, lipopolysaccharide, pro-inflammatory cytokines

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3032 Motif Search-Aided Screening of the Pseudomonas syringae pv. Maculicola Genome for Genes Encoding Tertiary Alcohol Ester Hydrolases

Authors: M. L. Mangena, N. Mokoena, K. Rashamuse, M. G. Tlou

Abstract:

Tertiary alcohol ester (TAE) hydrolases are a group of esterases (EC 3.1.1.-) that catalyze the kinetic resolution of TAEs and as a result, they are sought-after for the production of optically pure tertiary alcohols (TAs) which are useful as building blocks for number biologically active compounds. What sets these enzymes apart is, the presence of a GGG(A)X-motif in the active site which appears to be the main reason behind their activity towards the sterically demanding TAEs. The genome of Pseudomonas syringae pv. maculicola (Psm) comprises a multitude of genes that encode esterases. We therefore, hypothesize that some of these genes encode TAE hydrolases. In this study, Psm was screened for TAE hydrolase activity using the linalyl acetate (LA) plate assay and a positive reaction was observed. As a result, the genome of Psm was screened for esterases with a GGG(A)X-motif using the motif search tool and two potential TAE hydrolase genes (PsmEST1 and 2, 1100 and 1000bp, respectively) were identified, PsmEST1 was amplified by PCR and the gene sequenced for confirmation. Analysis of the sequence data with the SingnalP 4.1 server revealed that the protein comprises a signal peptide (22 amino acid residues) on the N-terminus. Primers specific for the gene encoding the mature protein (without the signal peptide) were designed such that they contain NdeI and XhoI restriction sites for directional cloning of the PCR products into pET28a. The gene was expressed in E. coli JM109 (DE3) and the clones screened for TAE hydrolase activity using the LA plate assay. A positive clone was selected, overexpressed and the protein purified using nickel affinity chromatography. The activity of the esterase towards LA was confirmed using thin layer chromatography.

Keywords: hydrolases, tertiary alcohol esters, tertiary alcohols, screening, Pseudomonas syringae pv., maculicola genome, esterase activity, linalyl acetate

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3031 Synthesis and Characterization of Fibrin/Polyethylene Glycol-Based Interpenetrating Polymer Networks for Dermal Tissue Engineering

Authors: O. Gsib, U. Peirera, C. Egles, S. A. Bencherif

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In skin regenerative medicine, one of the critical issues is to produce a three-dimensional scaffold with optimized porosity for dermal fibroblast infiltration and neovascularization, which exhibits high mechanical properties and displays sufficient wound healing characteristics. In this study, we report on the synthesis and characterization of macroporous sequential interpenetrating polymer networks (IPNs) combining skin wound healing properties of fibrin with the excellent physical properties of polyethylene glycol (PEG). Fibrin fibers serve as a provisional biologically active network to promote cell adhesion and proliferation while PEG provides the mechanical stability to maintain the entire 3D construct. After having modified both PEG and Serum Albumin (used for promoting enzymatic degradability) by adding methacrylate residues (PEGDM and SAM, respectively), Fibrin/PEGDM-SAM sequential IPNs were synthesized as follows: Macroporous sponges were first produced from PEGDM-SAM hydrogels by a freeze-drying technique and then rehydrated by adding the fibrin precursors. Environmental Scanning Electron Microscopy (ESEM) and Confocal Laser Scanning Microscopy (CLSM) were used to characterize their microstructure. Human dermal fibroblasts were cultivated during one week in the constructs and different cell culture parameters (viability, morphology, proliferation) were evaluated. Subcutaneous implantations of the scaffolds were conducted on five-week old male nude mice to investigate their biocompatibility in vivo. We successfully synthesized interconnected and macroporous Fibrin/PEGDM-SAM sequential IPNs. The viability of primary dermal fibroblasts was well maintained (above 90%) after 2 days of culture. Cells were able to adhere, spread and proliferate in the scaffolds suggesting the suitable porosity and intrinsic biologic properties of the constructs. The fibrin network adopted a spider web shape that covered partially the pores allowing easier cell infiltration into the macroporous structure. To further characterize the in vitro cell behavior, cell proliferation (EdU incorporation, MTS assay) is being studied. Preliminary histological analysis of animal studies indicated the persistence of hydrogels even after one-month post implantation and confirmed the absence of inflammation response, good biocompatibility and biointegration of our scaffolds within the surrounding tissues. These results suggest that our Fibrin/PEGDM-SAM IPNs could be considered as potential candidates for dermis regenerative medicine. Histological analysis will be completed to further assess scaffold remodeling including de novo extracellular matrix protein synthesis and early stage angiogenesis analysis. Compression measurements will be conducted to investigate the mechanical properties.

Keywords: fibrin, hydrogels for dermal reconstruction, polyethylene glycol, semi-interpenetrating polymer network

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3030 In Vitro Antibacterial Activity of Some Medicinal Plants Against Biofilm-Forming Methicillin-Resistant Staphylococcus aureus

Authors: Tesleem Adewale Ibrahim

Abstract:

Introduction: The prevalence of methicillin-resistant Staphylococcus aureus (MRSA) has been slowly rising in Nigeria for the past few decades. Therefore, novel classes of antibiotics are indispensable to combat the increased incidence of newly emerging multidrug-resistant bacteria like MRSA. Plants have been commonly used in popular medicine of most cultures for the treatment of disease. The in vitro antibacterial activity of some Nigerian common medicinal plants used in traditional medicine has been reported. The aim of this study was to investigate the antibacterial and anti-biofilm of these native plants (Entada abysinnica (leaves), Croton macrostachyus (leaves), Bridelia speciosa (seeds, bark), and Aframomum melegueta (leaves, seeds, and stem) collected in Southwestern Nigeria against a panel of seven biofilm-forming MRSA. Methods: Minimum inhibitory concentrations (MIC) and minimum bactericidal concentrations (MBC) of the plant extracts against MRSA were determined by the broth dilution method, and the anti-biofilm assay of the most potent plant extract was performed. Result: The results revealed that, of the four plants, water extracts of leaves of Entada abysinnica, leaves of Croton macrostachyus, seeds and bark Bridelia speciosa, and seeds of Aframomum melegueta exhibited significant antibacterial activity. Based on the MIC/MBC ratio, the extracts of these plants were determined to be bacteriostatic in nature. Anti-biofilm assay showed that the extract of seeds of Aframomum melegueta and leaves of Croton macrostachyus fairly inhibited the growth of MRSA in the preformed biofilm matrix. Conclusion: These four medicinal plant species may represent a source of alternative drugs derived from plant extracts based on folklore use and ethnobotanical knowledge from southwest Nigeria.

Keywords: extract, MRSA, antibacterial, biofilm, medicinal plants

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3029 Chaotic Representation: Translating Gender in Cantonese Opera Performances

Authors: Kar Yue Chan

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Cantonese opera is a valuable heritage originated from South China, and started to span its influence across the area to Hong Kong, and became extremely popular back in the 1950s to the 1970s. It has also been honoured and recognized as one treasurable item on the Intangible Cultural Heritage of Humanity on the Representative List of UNESCO since 2009. A certain level of difficulty is encountered when one identifies the gender roles and representations from a usual performance of Cantonese opera, as conventional practices of Cantonese opera display to the audience that many of the male hero roles are played by female upon the prior knowledge of all audience, and it is understandable for them as well because in the past there were insufficient male actors and performers on the market. Female actresses, in some senses, are more capable to sing near-male voices, and their appearances in heroic operatic attires are more appreciated by general audience. Therefore, perspectives of 'feminine representation' and the 'Reception Theory' in literature are conducive to analyzing such phenomenon. In spite of some 'normal' performances with romantic love stories or historical accounts involving often a talented intellectual and a beautiful wise lady (in Chinese caizi jiaren 才子佳人), in which the male role is actually male and the female role is actually female, there have still been some opera titles specifically manifesting these extreme gender associations by putting together displacement of gender roles in the same performance in view of such chaotic complication. On top of all other factors, translators dealing with any operatic texts face plenty of challenges upon transferring Cantonese operatic performances into English. It is found that translators need to deal with cultural elements embedded in the lyrics; the form (which is as delicate as those deriving from classical Chinese poetry); the gender misplacements that affect the mood and tone of the lyrics that much when they are in the process of translating. Some lyrics and tunes are specifically designed for a particular gender role to perform, while some others are more generic; both of which require different and specific translation strategies. After scrutinizing the various sources of reference, readers of this paper should be well informed of a significance which lies in the refined nature of the poetic form and content that signifies in the way the distinguished gender voice segregation of the discourse from which the lyrics are derived, and definitely also through the on-stage performability aspect of the task. In order to produce a relatively short and concise translated version which fits performance needs, all of the above factors will be looked at in this paper with relevant examples and analysis.

Keywords: Cantonese opera, translation, chaotic gender, performance

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3028 Ending the Gender Gap in Educational Leadership: A U.S. Goal for a Balanced Administration by 2030

Authors: S. Dodd

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This presentation examines the gender gap in leadership positions at colleges and universities within the United States. Despite the fact that women now outnumber men in earning doctorate degrees, women continue to hold far fewer positions of educational leadership, and still, earn less money than men do at every level. Considering the lack of female representation in positions of leadership, there are clearly outside variables preventing women from attaining these positions, despite their educational attainment. Following this study, the American Council on Education (ACE) set a goal to achieve an equal percentage of females holding college presidency positions by the year 2030. This goal is particularly ambitious, especially when considering the gender disparity at all ranks in higher education. Men still hold nearly 70% of all full professorships at degree-granting institutions. Even when women are equally represented in numbers, men typically hold a higher rank and are more likely to be tenured. Across all four-year colleges and universities in the United States, men earn more money than women at every rank and in every discipline. There are over twice as many men than women represented on governing boards, who help formed and uphold campus policies. The fact that the low percentage of female presidents has remained static for many years deepens the challenge for the ACE. Although emphasizing the need to create greater opportunities for women in educational administration is admirable, it is difficult to simplify the social forces that create and uphold the status quo of male leadership. When aiming to ensure 'women' hold 50% of all college presidency positions, it is important to consider how the intersections of race, social class, and other factors also correlate with lower job status. This presentation explores how gendered notions of leadership begin in a child’s early years and are carried into future careers, and how these conceptualizations impact the creation and upholding of educational policies at every academic level. Current research that emphasizes the importance establishing a bottom-up approach to a gender equity infrastructure for children early in their educational careers will be discussed. A top-down approach starting with female college presidents is incomplete and insufficient if the mindsets of the youth who will one day be entering those institutions of higher education are not also taken into consideration. Although ACE has established this lofty goal for female college presidencies by the year 2030, a road map for this will ensue, has not yet been provided. The talent pool of women who are educated and experienced for such positions is vast, but acknowledging the social barriers existing for women in these positions will be crucial to making the changes necessary for these leadership opportunities to be long lasting and successful.

Keywords: equity, higher education, leadership, women

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3027 Role of Molecular Changes and Immunohistochamical in Early Detection of Colon Cancer

Authors: Fatimah Alhomaid

Abstract:

The present study was planned to investigate the role of molecular changes and immunohistochemical in early detection of colon cancer in Saudi patients. Our results were carried out on 48 patients colon cancer. We obtained our data from laboratory in King Khalid university hospital. The specimens were taken (48) patients with colon cancer 34 male and 14 female and 2 control. The average age of varied from 37-85 years. The tumor was diagnosed as I in tow patients (male and female) and grade 2 in 42 patients (29 male and 13 female) while the grade 3 in 4 patients (all males). The specimens were processed for haematoxylin and eosin staining , immunohistochemical technique and flow cytometry analysis. Our study noted that most patients had adenocarcinoma which characterized by presence of signet-ring cells were very clear in advanced patients of adenocarcinoma. Our sections in adenocarcinoma in grade 2 and stage 3 had an increase in signet ring cells,an increase in the acini of glands and an increase in number of lymphocytes which spread to the muscularis layer. With advancing the disease, there were haemorge in blood and increase in lymphocytes and increase number of nuclei in the tubular glands. Our study was carried on 48 patients, immunohistochemical diagnosis (CK20,PCNA,P53) and the analysis of DNA content by flow cytometry technique. Our study indicated that the presence of correlation between the immunohistochemical analysis for P53 and the grades. The reaction of P53 appeared as strong in nucleus in grades &stage 3 and appeared in other sections as dark brown pigment. Our study indicated that the absence of correlation between the immunohistochemical analysis for pcan and the grades. In our sections, there were strong reactions in the more 80% of nuclei in grade 1& stage 2. Our study indicated that the presence of correlation between the immunohistochemical analysis for CK20 and the grades. Our results indicated the presence of positive reaction in cytoplasm varied from weak to moderate in grade 3 & stage 4. Concerning the Flow cytometry technique our results indicated that the presence of correlation between the DNA and different stages of colon cancer.

Keywords: DNA-CK20, PCNA, P53, colon cancer

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3026 Meditation Based Brain Painting Promotes Foreign Language Memory through Establishing a Brain-Computer Interface

Authors: Zhepeng Rui, Zhenyu Gu, Caitilin de Bérigny

Abstract:

In the current study, we designed an interactive meditation and brain painting application to cultivate users’ creativity, promote meditation, reduce stress, and improve cognition while attempting to learn a foreign language. User tests and data analyses were conducted on 42 male and 42 female participants to better understand sex-associated psychological and aesthetic differences. Our method utilized brain-computer interfaces to import meditation and attention data to create artwork in meditation-based applications. Female participants showed statistically significantly different language learning outcomes following three meditation paradigms. The art style of brain painting helped females with language memory. Our results suggest that the most ideal methods for promoting memory attention were meditation methods and brain painting exercises contributing to language learning, memory concentration promotion, and foreign word memorization. We conclude that a short period of meditation practice can help in learning a foreign language. These findings provide new insights into meditation, creative language education, brain-computer interface, and human-computer interactions.

Keywords: brain-computer interface, creative thinking, meditation, mental health

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3025 Gender Identity: Omani College Students Negotiate Their Cultural Expectations

Authors: Mohammed Alkharusi

Abstract:

This study addresses issues of gender identity faced by female and male Omani students studying at educational higher institutions. The study interviewed 16 male and female students to understand how cultural expectations of gender influence these students’ communication, and as a result how these students negotiate their gender identity to facilitate communication practices (or not) with the opposite sex. The context, focus, and theoretical underpinnings of the study are presented. Given that the researcher is also an Omani Arab, methodological and ethical challenges (e.g., recruiting and engaging with participants, and conducting semi-structured face-to-face interviews) will be discussed reflexively. The analysis found that students continued to following cultural expectations. They kept minimum interaction with the opposite sex that was illustrated by preferring to work with the same sex in group assignments only, avoiding sitting alone with the opposite sex, and not participating in academic activities. In the social context, the students started negotiating their gender identity and adopted communication practices that facilitated their social communication with the opposite sex. For example, they accepted to work with the opposite sex in different social mixed activities. In conclusion, students desired to maintain their cultural expectations but adopted certain communication practices to interact with the opposite sex.

Keywords: communication, cultural expectations, gender, identity, negotiation

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3024 Coping Strategies of Female English Teachers and Housewives to Face the Challenges Associated to the COVID-19 Pandemic Lockdown

Authors: Lisseth Rojas Barreto, Carlos Muñoz Hernández

Abstract:

The COVID-19 pandemic led to many abrupt changes, including a prolonged lockdown, which brought about work and personal challenges to the population worldwide. Among the most affected populations are women who are workers and housewives at the same time, and especially those who are also parenting. These women were faced with the challenge to perform their usual varied roles during the lockdown from the same physical space, which inevitably had strong repercussions for each of them. This paper will present some results of a research study whose main objective was to examine the possible effects that the COVID-19 pandemic lockdown may have caused in the work, social, family, and personal environments of female English teachers who are also housewives and, by extension in the teaching and learning processes that they lead. Participants included five female English language teachers of a public foreign language school, they are all married, and two of them have children. Similarly, we examined some of the coping strategies these teachers used to tackle the pandemic-related challenges in their different roles, especially those used for their language teaching role; coping strategies are understood as a repertoire of behaviors in response to incidents that can be stressful for the subject, possible challenging events or situations that involve emotions with behaviors and decision-making of people which are used in order to find a meaning or positive result (Lazarus &Folkman, 1986) Following a qualitative-case study design, we gathered the data through a survey and a focus group interview with the participant teachers who work at a public language school in southern Colombia. Preliminary findings indicate that the circumstances that emerged as a result of the pandemic lockdown affected the participants in different ways, including financial, personal, family, health, and work-related issues. Among the strategies that participants found valuable to deal with the novel circumstances, we can highlight the reorganization of the household and work tasks and the increased awareness of time management for the household, work, and leisure. Additionally, we were able to evidence that the participants faced the circumstances with a positive view. Finally, in order to cope with their teaching duties, some participants acknowledged their lack of computer or technology literacy in order to deliver their classes online, which made them find support from their students or more knowledgeable peers to cope with it. Others indicated that they used strategies such as self-learning in order to get acquainted and be able to use the different technological tools and web-based platforms available.

Keywords: coping strategies, language teaching, female teachers, pandemic lockdown

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3023 Satureja bachtiarica Bunge Induce Apoptosis via Mitochondrial Intrinsic Pathway and G1 Cell Cycle Arrest

Authors: Hamed Karimian, Noraziah Nordin, Mohamad Ibrahim Noordin, Syam Mohan, Mahboubeh Razavi, Najihah Mohd Hashim, Happipah Mohd Ali

Abstract:

Satureja bachtiarica Bunge is a perennial medicinal plant belonging to the Lamiaceae family and endemic species in Iran. Satureja bachtiarica Bunge with the local name of Marzeh koohi is edible vegetable use as flavoring agent, anti-bacterial and to relieve cough and indigestion. In this study, the anti-cancer effect of Satureja bachtiarica Bunge on the MDA-MB-231 cell line as an Breast cancer cell model has been analyzed for the first time. Satureja bachtiarica Bunge was extracted using different solvents in the order of increasing polarity. Cytotoxicity activity of hexane extract of Satureja bachtiarica Bunge (SBHE) was observed using MTT assay. Acridine orange/Propidium iodide staining was used to detect early apoptosis; Annexin-V-FITC assay was carried out to observe the detection of cell-surface Phosphatidylserine (PS), with Annexin-Vserving as a marker for apoptotic cells. Caspase 3/7, 8 and-9 assays showed significantly activation of caspase-9 where lead intrinsic mitochondrial pathway. Bcl-2/Bax expressions and cell cycle arrest were also investigated. SBHE had exhibited significantly higher cytotoxicity against MDA-MB-231 Cell line compare to other cell lines. A significant increase in chromatin condensation in the cell nucleus was observed by fluorescence analysis. Treatment of MDA-MB-231 cells with SBHE encouraged apoptosis, by down-regulating Bcl-2 and up-regulating Bax, which lead the activation of caspase 9. Moreover, SBHE treatment significantly arrested MDA-MB-231 cells in the G1 phase. Together, the results presented in this study demonstrated that SBHE inhibited the proliferation of MDA-MB-231 cells, leading cell cycle arrest and programmed cell death, which was confirmed to be through the mitochondrial pathway.

Keywords: Satureja bachtiarica Bunge, MDA-MB-231, apoptosis, annexin-V, cell cycle

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3022 Association Analysis of Putative Loci with Coronary Artery Disease

Authors: Asma Naseer Cheema, Attya Bhatti, Jabar Ali, John Peter

Abstract:

Background: High cholesterol levels, endothelial dysfunction, inefficient coagulation cascade and hyper inflammatory response all are the basis of coronary artery disease (CAD). Several studies are carried out to see the genetic influence of these factors on disease outcome. Objective: The objective of our study was to see the association of 10 putative loci with coronary artery disease in our population. Materials & Methods: We screened our population for 10 putative loci of CAD showing significant association (p < 5x10-8) with candidate genes (regulating the cholesterol metabolism, endothelial function, coagulation cascade and inflammatory response of body). Hardy-Weinberg equilibrium and linkage disequilibrium in cases and controls s were estimated separately. Approximately 5-10 ng of dried DNA in 384 well plate format was used to genotype each sample on the Sequenom iPLEX assay at University of Pittsburgh Genomics and Proteomics Core Laboratories. It was built on single-base primer extension with the MALDI-TOF MS detection possessing high sensitivity and specificity. The SNPs were genotyped through Taqman assay. Hardy Weinberg test was applied. The 10 SNPs were selected as genetic markers for this study (rs579459, rs1561198, rs2954029, rs1122608, rs17114036, rs9515203, rs10947789, rs7173743, rs2895811, rs2075650). Results: Mean age of the patient was 52 ± 11 years. Blood pressure and positive family history was found a significant risk factor for CAD. None of the selected SNPs showed significant association with coronary artery disease in our population (p>0.05). Conclusion: rs579459, rs1561198, rs2954029, rs1122608, rs17114036, rs9515203, rs10947789, rs7173743, rs2895811, rs2075650 are not significant genetic markers for CAD in our population.

Keywords: CAD, genetic markers, loci, risk factors

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3021 Paradigm Shift in Classical Drug Research: Challenges to Mordern Pharmaceutical Sciences

Authors: Riddhi Shukla, Rajeshri Patel, Prakruti Buch, Tejas Sharma, Mihir Raval, Navin Sheth

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Many classical drugs are claimed to have blood sugar lowering properties that make them valuable for people with or at high risk of type 2 diabetes. Vijaysar (Pterocarpus marsupium) and Gaumutra (Indian cow urine) both have been shown antidiabetic property since primordial time and both shows synergistic effect in combination for hypoglycaemic activity. The study was undertaken to investigate the hypoglycaemic and anti-diabetic effects of the combination of Vijaysar and Gaumutra which is a classical preparation mentioned in Ayurveda named as Pramehari ark. Rats with Type 2 diabetes which is induced by streptozotocin (STZ, 35mg/kg) given a high-fat diet for one month and compared with normal rats. Diabetic rats showed raised level of body weight, triglyceride (TG), total cholesterol, HDL, LDL, and D-glucose concentration and other serum, cardiac and hypertrophic parameters in comparison of normal rats. After treatment of different doses of drug the level of parameters like TG, total cholesterol, HDL, LDL, and D-glucose concentration found to be decreased in standard as well as in treatment groups. In addition treatment groups also found to be decreased in the level of serum markers, cardiac markers, and hypertrophic parameters. The findings demonstrated that Pramehari ark prevented the pathological progression of type 2 diabetes in rats.

Keywords: cow urine, hypoglycemic effect, synergic effect, type 2 diabetes, vijaysar

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3020 Cryotopic Macroporous Polymeric Matrices for Regenerative Medicine and Tissue Engineering Applications

Authors: Archana Sharma, Vijayashree Nayak, Ashok Kumar

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Three-dimensional matrices were fabricated from blend of natural-natural polymers like carrageenan-gelatin and synthetic -natural polymers such as PEG- gelatin (PEG of different molecular weights (2,000 and 6,000) using two different crosslinkers; glutaraldehyde and EDC-NHS by cryogelation technique. Blends represented a feasible approach to design 3-D scaffolds with controllable mechanical, physical and biochemical properties without compromising biocompatibility and biodegradability. These matrices possessed interconnected porous structure, good mechanical strength, biodegradable nature, constant swelling kinetics, ability to withstand high temperature and visco-elastic behavior. Hemocompatibility of cryogel matrices was determined by coagulation assays and hemolytic activity assay which demonstrated that these cryogels have negligible effects on coagulation time and have excellent blood compatibility. In vitro biocompatibility (cell-matrix interaction) inferred good cell adhesion, proliferation, and secretion of ECM on matrices. These matrices provide a microenvironment for the growth, proliferation, differentiation and secretion of ECM of different cell types such as IMR-32, C2C12, Cos-7, rat bone marrow derived MSCs and human bone marrow MSCs. Hoechst 33342 and PI staining also confirmed that the cells were uniformly distributed, adhered and proliferated properly on the cryogel matrix. An ideal scaffold used for tissue engineering application should allow the cells to adhere, proliferate and maintain their functionality. Neurotransmitter analysis has been done which indicated that IMR-32 cells adhered, proliferated and secreted neurotransmitters when they interacted with these matrices which showed restoration of their functionality. The cell-matrix interaction up to molecular level was also evaluated so to check genotoxicity and protein expression profile which indicated that these cryogel matrices are non-genotoxic and maintained biofunctionality of cells growing on these matrices. All these cryogels, when implanted subcutaneously in balb/c mice, showed no adverse systemic or local toxicity effects at implantation site. There was no significant increase in inflammatory cell count has otherwise been observed after scaffold implantation. These cryogels are supermacroporous and this porous structure allows cell infiltration and proliferation of host cells. This showed the integration and presence of infiltrated cells into the cryogel implants. Histological analysis confirmed that the implanted cryogels do not have any adverse effect in spite of host immune system recognition at the site of implantation, on its surrounding tissues and other vital host organs. In vivo biocompatibility study after in vitro biocompatibility analysis has also concluded that these synthesized cryogels act as important biological substitutes, more adaptable and appropriate for transplantation. Thus, these cryogels showed their potential for soft tissue engineering applications.

Keywords: cryogelation, hemocompatibility, in vitro biocompatibility, in vivo biocompatibility, soft tissue engineering applications

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3019 Prevalence of Metabolic Syndrome According to Different Criteria in Population over 20 Years Old in Ahvaz

Authors: Armaghan Moravej Aleali, Hajieh Shahbazian, Seyed Mahmoud Latifi, Leila Yazdanpanah

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Objective: Metabolic syndrome or insulin resistance syndrome or syndrome X is a collection of abdominal obesity, hypertension, glucose intolerance and lipid abnormalities (elevated triglycerides, elevated LDL, and decrease the amount of HDL). That increases the incidence of diabetes and risk of cardiovascular disease. The aim of this study is to investigate the prevalence of metabolic syndrome in people over 20 years of Ahvaz according to IDF, ATPIII, Harmonized I and Harmonized II. Material & Methods: A cross-sectional study with a random cluster sampling in six health centers in Ahvaz was done. After obtaining informed consent, questionnaire for each person filled up including demographic data and examinations, including blood pressure in sitting position, weight, height, waist circumference, and waist circumference measurement. Results: From all participating 912 people, (434 (2/47%) male and 478 (2/52%) female) were evaluated. Mean age was 42/27± 14years (44/2±14/26 for male and 40/5±13/5 for female). Prevalence of metabolic syndrome was 22/8%, 28/4%, 30/9% and 16/9% according to ATPIII, IDF, Harmonized I and Harmonized II criteria respectively and increased with age in both sexes. IDF and Harmonized I had most kappa coordination (0/94). Conclusion: The results show a high prevalence of metabolic syndrome in Ahvaz. So, identification of the risk factors should be attempted to prevent metabolic syndrome.

Keywords: metabolic syndrome, IDF, ATP III, prevalence

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3018 Psychosocial Correlates of Sexual Violence Among Students in Higher Institutions in Cameroon

Authors: Agbor Ekama Prisca Anne

Abstract:

Current data on the prevalence and psychosocial correlates of sexual violence in the Cameroon is lacking, with the most recent sexual abuse and violence survey dating back to 2001. The current study sought to identify what proportion of University students have experienced sexual violence, if there are sex differences in exposure to different forms of sexual violence, and to what extent different forms of sexual violence are associated with adverse psychosocial outcomes. A nationally representative sample of University students (N = 1,020) completed self-report measures of history of sexual violence and mental health. Approximately one-in-three (34.4%) students experienced some form of sexual violence, including 14.8% who were sexually assaulted (raped) and 31.1% who were sexually harassed. Female students were significantly more likely than men to have experienced all forms of sexual violence (ps < .001), with the exception of sexual assault by teachers or guardian. All forms of sexual violence were associated with an increased likelihood of serious mental health problems, with sexual assault by a teacher associated with several other psychosocial outcomes in life, including education achievement, and behavior disorder. Sexual violence is a common experience in the general population and female students are disproportionately affected (1-in-2 girls versus 1-in-5 boys). Additional resources to increase mental health care among survivors of sexual violence is urgently needed.

Keywords: psychosocial, effects sexual, violence, females, students

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3017 Inhibition of Influenza Replication through the Restrictive Factors Modulation by CCR5 and CXCR4 Receptor Ligands

Authors: Thauane Silva, Gabrielle do Vale, Andre Ferreira, Marilda Siqueira, Thiago Moreno L. Souza, Milene D. Miranda

Abstract:

The exposure of A(H1N1)pdm09-infected epithelial cells (HeLa) to HIV-1 viral particles, or its gp120, enhanced interferon-induced transmembrane protein (IFITM3) content, a viral restriction factor (RF), resulting in a decrease in influenza replication. The gp120 binds to CCR5 (R5) or CXCR4 (X4) cell receptors during HIV-1 infection. Then, it is possible that the endogenous ligands of these receptors also modulate the expression of IFITM3 and other cellular factors that restrict influenza virus replication. Thus, the aim of this study is to analyze the role of cellular receptors R5 and X4 in modulating RFs in order to inhibit the replication of the influenza virus. A549 cells were treated with 2x effective dose (ED50) of endogenous R5 or X4 receptor agonists, CCL3 (20 ng/ml), CCL4 (10 ng/ml), CCL5 (10 ng/ml) and CXCL12 (100 ng/mL) or exogenous agonists, gp120 Bal-R5, gp120 IIIB-X4 and its mutants (5 µg/mL). The interferon α (10 ng/mL) and oseltamivir (60 nM) were used as a control. After 24 h post agonists exposure, the cells were infected with virus influenza A(H3N2) at 2 MOI (multiplicity of infection) for 1 h. Then, 24 h post infection, the supernatant was harvested and, the viral titre was evaluated by qRT-PCR. To evaluate IFITM3 and SAM and HD domain containing deoxynucleoside triphosphate triphosphohydrolase 1 (SAMHD1) protein levels, A549 were exposed to agonists for 24 h, and the monolayer was lysed with Laemmli buffer for western blot (WB) assay or fixed for indirect immunofluorescence (IFI) assay. In addition to this, we analyzed other RFs modulation in A549, after 24 h post agonists exposure by customized RT² Profiler Polymerase Chain Reaction Array. We also performed a functional assay in which SAMHD1-knocked-down, by single-stranded RNA (siRNA), A549 cells were infected with A(H3N2). In addition, the cells were treated with guanosine to assess the regulatory role of dNTPs by SAMHD1. We found that R5 and X4 agonists inhibited influenza replication in 54 ± 9%. We observed a four-fold increase in SAMHD1 transcripts by RFs mRNA quantification panel. After 24 h post agonists exposure, we did not observe an increase in IFITM3 protein levels through WB or IFI assays, but we observed an upregulation up to three-fold in the protein content of SAMHD1, in A549 exposed to agonists. Besides this, influenza replication enhanced in 20% in cell cultures that SAMDH1 was knockdown. Guanosine treatment in cells exposed to R5 ligands further inhibited influenza virus replication, suggesting that the inhibitory mechanism may involve the activation of the SAMHD1 deoxynucleotide triphosphohydrolase activity. Thus, our data show for the first time a direct relationship of SAMHD1 and inhibition of influenza replication, and provides perspectives for new studies on the signaling modulation, through cellular receptors, to induce proteins of great importance in the control of relevant infections for public health.

Keywords: chemokine receptors, gp120, influenza, virus restriction factors

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3016 Learning Performance of Sports Education Model Based on Self-Regulated Learning Approach

Authors: Yi-Hsiang Pan, Ching-Hsiang Chen, Wei-Ting Hsu

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The purpose of this study was to compare the learning effects of the sports education model (SEM) to those of the traditional teaching model (TTM) in physical education classes in terms of students learning motivation, action control, learning strategies, and learning performance. A quasi-experimental design was utilized in this study, and participants included two physical educators and four classes with a total of 94 students in grades 5 and 6 of elementary schools. Two classes implemented the SEM (n=47, male=24, female=23; age=11.89, SD=0.78) and two classes implemented the TTM (n=47, male=25, female=22, age=11.77; SD=0.66). Data were collected from these participants using a self-report questionnaire (including a learning motivation scale, action control scale, and learning strategy scale) and a game performance assessment instrument, and multivariate analysis of covariance was used to conduct statistical analysis. The findings of the study revealed that the SEM was significantly better than the TTM in promoting students learning motivation, action control, learning strategies, and game performance. It was concluded that the SEM could promote the mechanics of students self-regulated learning process, and thereby improve students movement performance.

Keywords: self-regulated learning theory, learning process, curriculum model, physical education

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3015 Gendered Effects on Productivity Gap Due to Information Asymmetry

Authors: Shruti Sengupta

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According to the nationally representative data, about 73% of India's rural workforce is engaged in agriculture. While women make significant contributions to total agriculture production, they contribute to about one-third in India. In terms of gender composition, about 80% of the female and 69% of the male workforce is engaged in agriculture in rural India. Still, it is common to find gender differences in plot management within the household. In the last two and half years, India's agri-food system has undergone several changes due to this pandemic, both the demand and supply side, making agriculture more information and knowledge-intensive. Therefore, this paper investigates, using a nationally representative sample, how information asymmetry affects the net returns per hectare of land between female and male farm managers. Empirical results show that information intensity has a significant positive effect on net farm returns per hectare. Results suggest that if females have the same access to technical information as their male counterparts, their farm income can go up by .96 pp compared to male-headed farms. Results also indicate that literate females have higher farm incomes than non-literate females. The study contributes to the literature by employing gender differentials in farm income due to the information gap.

Keywords: agriculture, gender, information asymmetry, farm income, social bias

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3014 An Antidiabetic Dietary Defence Weapon: Oats and Milk Based Probiotic Fermented Product

Authors: Rameshwar Singh Seema

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In today’s world where diabetes has become an epidemic, our aim was to potentiate the effect of probiotics by integrating probiotics with cereals to formulate composite foods using Lactobacillus rhamnosus GG (LGG) and Lactobacillus casei NCDC19 against type 2 diabetes. After optimizing the product by Response Surface Methodology, it was studied for their effect on induction and progression of type 2 diabetes in HFD-fed Wistar rats. After 9 weeks study, best results were shown by the group fed with oat and milk based product fermented with LGG and L. casei NCDC19 which resulted in a significant decrease in blood glucose, HBA1c, improved OGTT, oxidative stress, cholesterol and triglycerides level during progression study of type 2 diabetes. During induction study also, there was significant reduction in blood glucose level, oxidative stress, cholesterol level and triglycerides level but slightly less as compared to progression study. Real time PCR gene expression studies were done for 5 genes (GLUT-4, IRS-2, ppar-γ, TNF-α, IL-6) whose expression is directly related to type 2 diabetes. The relative fold change expression was increased in case of GLUT-4, IRS-2, ppar-γ and decreased in case of TNF-α and IL-6 during both induction and progression study of diabetes but more significantly during progression study. Hence it was concluded that oat and milk based probiotic fermented product showed the synergistic effect of probiotics and oats especially in case of progression of type 2 diabetes. The benefits of these probiotic formulations may be further validated by clinical trials.

Keywords: type 2 diabetes, LGG, L.casei NCDC19, food science

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3013 Nurturing Green Creativity in Women Intrapreneurs through Green HRM: Testing Moderated Mediation Model: A Step Towards Saudi Vision 2030

Authors: Tahira Iram, Ahmad Raza Bilal

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In 2016, the Kingdom of Saudi Arabia (KSA) initiated Saudi Vision 2030, an ambitious plan to lessen the country's dependency on fossil fuels and increase economic diversification. The Vision 2030 framework strives to establish a thriving economy, a vibrant society, and an ambitious nation. This study aims to investigate the role of green service innovation (SI) and green work engagement (WE) in mediating the nexus between green HRM and green creativity (GC) under the conditional role of spiritual leadership (SL). A survey was done of 300 female intrepreneurs working in the organization within Saudi Arabia. This study has collected data via a stratified random sampling technique. The framework was tested using PLS-SEM software. The findings reveal that WE fully intervenes in the nexus between green HRM and GC. Moreover, SL positively moderates the nexus between green HRM and SI. Thus based on findings, it is recommended that female intrapreneurs prioritize environmentally responsible operations to gain and sustain a competitive edge over rivals in the Saudi competitive market.

Keywords: green HRM, spiritual leadership, Vision 2030, women intrapreneurs, green service innovation behavior, green creativity

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