Search results for: cell biology
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 3917

Search results for: cell biology

737 Hesperidin through Acting on Proliferating Cell Nuclear Antigen and Follicle Stimulating Hormone Receptor Expression Decreased Ovarian Toxicity Induced by Malathion

Authors: Mahnaz Zarein, Hamed Shoorei

Abstract:

Background: Malathion is one of the most toxic agents widely used in agriculture throughout the world. This agent has adverse effects on the functions of multiple organs such as the reproductive system in both male and female genders. On the one hand, daily use of antioxidant supplementations such as hesperidin is capable to neutralize the deleterious impacts of malathion. Therefore, in this experimental study, the protective effects of hesperidin against ovarian toxicity induced by malathion were investigated. Material & Methods: Balb/c adult mice (n=32) were randomly divided into 4 groups including 1) the control group, treated with normal saline, 2) the Mal group, treated with 30mg/kg malathion, daily for 1 month, 3) Mal + Hes group, treated with 20 mg/kg malathion and 20 mg/kg hesperidin, daily for 1 month, and 5) Hes group, treated with 20 mg/kg hesperidin. At the end of the experimental period, mice were anesthetized and their drops of blood were collected to the assessment of some hormones such as LH, FSH, E2, and P4. Also, the right ovaries were used to H&E staining, and the left ovaries were used for IHC staining (PCNA and FSHR). Results: Histopathological assessments showed that the number of follicles, i.e. primordial, primary, and secondary, significantly decreased, while the atretic follicle counts remarkably increased compared to the control group (p<0.05). Hormonal levels revealed that the production of all mentioned hormones decreased in the Mal group in comparison with the control group (p<0.05). The expression of PCNA, as a proliferative marker, and FHSR, as a marker showing maturation, decreased when mice received malathion compared to the control group (p<0.05). Interestingly, treatment with hesperidin significantly neutralized the adverse effects of malathion on all mentioned parameters. Conclusion: Daily use of antioxidant supplementations such as hesperidin could alleviate the ovarian toxicity induced by malathion.

Keywords: malathion, ovary, antioxidant hesperidin, FSHR PCNA, ovary

Procedia PDF Downloads 65
736 In vitro Effects of Salvia officinalis on Bovine Spermatozoa

Authors: Eva Tvrdá, Boris Botman, Marek Halenár, Tomáš Slanina, Norbert Lukáč

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In vitro storage and processing of animal semen represents a risk factor to spermatozoa vitality, potentially leading to reduced fertility. A variety of substances isolated from natural sources may exhibit protective or antioxidant properties on the spermatozoon, thus extending the lifespan of stored ejaculates. This study compared the ability of different concentrations of the Salvia officinalis extract on the motility, mitochondrial activity, viability and reactive oxygen species (ROS) production by bovine spermatozoa during different time periods (0, 2, 6 and 24 h) of in vitro culture. Spermatozoa motility was assessed using the Computer-assisted sperm analysis (CASA) system. Cell viability was examined using the metabolic activity MTT assay, the eosin-nigrosin staining technique was used to evaluate the sperm viability and ROS generation was quantified using luminometry. The CASA analysis revealed that the motility in the experimental groups supplemented with 0.5-2 µg/mL Salvia extract was significantly lower in comparison with the control (P<0.05; Time 24 h). At the same time, a long-term exposure of spermatozoa to concentrations ranging between 0.05 µg/mL and 2 µg/mL had a negative impact on the mitochondrial metabolism (P<0.05; Time 24 h). The viability staining revealed that 0.001-1 µg/mL Salvia extract had no effects on bovine male gametes, however 2 µg/mL Salvia had a persisting negative effect on spermatozoa (P<0.05). Furthermore 0.05-2 µg/mL Salvia exhibited an immediate ROS-promoting effect on the sperm culture (P>0.05; Time 0 h and 2 h), which remained significant throughout the entire in vitro culture (P<0.05; Time 24 h). Our results point out to the necessity to examine specific effects the biomolecules present in Salvia officinalis may have individually or collectively on the in vitro sperm vitality and oxidative profile.

Keywords: bulls, CASA, MTT test, reactive oxygen species, sage, Salvia officinalis, spermatozoa

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735 Real-Time Quantitative Polymerase Chain Reaction Assay for the Detection of microRNAs Using Bi-Directional Extension Sequences

Authors: Kyung Jin Kim, Jiwon Kwak, Jae-Hoon Lee, Soo Suk Lee

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MicroRNAs (miRNA) are a class of endogenous, single-stranded, small, and non-protein coding RNA molecules typically 20-25 nucleotides long. They are thought to regulate the expression of other genes in a broad range by binding to 3’- untranslated regions (3’-UTRs) of specific mRNAs. The detection of miRNAs is very important for understanding of the function of these molecules and in the diagnosis of variety of human diseases. However, detection of miRNAs is very challenging because of their short length and high sequence similarities within miRNA families. So, a simple-to-use, low-cost, and highly sensitive method for the detection of miRNAs is desirable. In this study, we demonstrate a novel bi-directional extension (BDE) assay. In the first step, a specific linear RT primer is hybridized to 6-10 base pairs from the 3’-end of a target miRNA molecule and then reverse transcribed to generate a cDNA strand. After reverse transcription, the cDNA was hybridized to the 3’-end which is BDE sequence; it played role as the PCR template. The PCR template was amplified in an SYBR green-based quantitative real-time PCR. To prove the concept, we used human brain total RNA. It could be detected quantitatively in the range of seven orders of magnitude with excellent linearity and reproducibility. To evaluate the performance of BDE assay, we contrasted sensitivity and specificity of the BDE assay against a commercially available poly (A) tailing method using miRNAs for let-7e extracted from A549 human epithelial lung cancer cells. The BDE assay displayed good performance compared with a poly (A) tailing method in terms of specificity and sensitivity; the CT values differed by 2.5 and the melting curve showed a sharper than poly (A) tailing methods. We have demonstrated an innovative, cost-effective BDE assay that allows improved sensitivity and specificity in detection of miRNAs. Dynamic range of the SYBR green-based RT-qPCR for miR-145 could be represented quantitatively over a range of 7 orders of magnitude from 0.1 pg to 1.0 μg of human brain total RNA. Finally, the BDE assay for detection of miRNA species such as let-7e shows good performance compared with a poly (A) tailing method in terms of specificity and sensitivity. Thus BDE proves a simple, low cost, and highly sensitive assay for various miRNAs and should provide significant contributions in research on miRNA biology and application of disease diagnostics with miRNAs as targets.

Keywords: bi-directional extension (BDE), microRNA (miRNA), poly (A) tailing assay, reverse transcription, RT-qPCR

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734 Effect of Zidovudine on Hematological and Virologic Parameters among Female Sex Workers Receiving Antiretroviral Therapy (ART) in North-Western Nigeria

Authors: N. M. Sani, E. D. Jatau, O. S. Olonitola, M. Y. Gwarzo, P. Moodley, N. S. Mujahid

Abstract:

Haemoglobin (HB) indicates anaemia level and by extension may reflect the nutritional level and perhaps the immunity of an individual. Some antiretroviral drugs like zidovudine are known to cause anaemia in People living with HIV/AIDS (PLWHA). A cross-sectional study using demographic data and blood specimen from 218 female commercial sex workers attending antiretroviral therapy (ART) clinics was conducted between December 2009 and July 2011 to assess the effect of zidovudine on haematologic and RNA viral load of female sex workers receiving antiretroviral treatment in north-western Nigeria. Anaemia is a common and serious complication of both HIV infection and its treatment. In the setting of HIV infection, anaemia has been associated with decreased quality of life, functional status, and survival. Antiretroviral therapy, particularly the highly active antiretroviral therapy (HAART), has been associated with a decrease in the incidence and severity of anaemia in HIV-infected patients who have received a HAART regimen for at least 1 year. In this study, result has shown that out of 218 patients, 26 with haemoglobin count between 5.1–10 g/dl were observed to have the highest viral load count of 300,000–350,000 copies/ml. It was also observed that most patients (190) with HB of 10.1–15.0 g/dl had viral load count of 200,000–250,000 copies/ml. An inverse relationship therefore exists, i.e. the lower the haemoglobin level, the higher the viral load count, even though the test statistics did not show any significance between the two (P=0.206). This shows that multivariate logistic regression analysis demonstrated that anaemia was associated with a CD4+ cell count below 50/µL in female sex workers with a viral load above 100,000 copies/mL who use zidovudine. Severe anaemia was less prevalent in this study population than in historical comparators; however, mild to moderate anaemia rates remain high. The study, therefore, recommends that hematological and virologic parameters be monitored closely in patients receiving first line ART regimen.

Keywords: anaemia, female sex worker, haemoglobin, Zidovudine

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733 Tumour Radionuclides Therapy: in vitro and in vivo Dose Distribution Study

Authors: Rekaya A. Shabbir, Marco Mingarelli, Glenn Flux, Ananya Choudhury, Tim A. D. Smith

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Introduction: Heterogeneity of dose distributions across a tumour is problematic for targeted radiotherapy. Gold nanoparticles (AuNPs) enhance dose-distributions of targeted radionuclides. The aim of this study is to demonstrate if tumour dose-distribution of targeted AuNPs radiolabelled with either of two radioisotopes (¹⁷⁷Lu and ⁹⁰Y) in breast cancer cells produced homogeneous dose distributions. Moreover, in vitro and in vivo studies were conducted to study the importance of receptor level on cytotoxicity of EGFR-targeted AuNPs in breast and colorectal cancer cells. Methods: AuNPs were functionalised with DOTA and OPPS-PEG-SVA to optimise labelling with radionuclide tracers and targeting with Erbitux. Radionuclides were chelated with DOTA, and the uptake of the radiolabelled AuNPs and targeted activity in vitro in both cell lines measured using liquid scintillation counting. Cells with medium (HCT8) and high (MDA-MB-468) EGFR expression were incubated with targeted ¹⁷⁷Lu-AuNPs for 4h, then washed and allowed to form colonies. Nude mice bearing tumours were used to study the biodistribution by injecting ¹⁷⁷Lu-AuNPs or ⁹⁰Y-AuNPs via the tail vein. Heterogeneity of dose-distribution in tumours was determined using autoradiography. Results: Colony formation (% control) was 81 ± 4.7% (HCT8) and 32 ± 9% (MDA-MB-468). High uptake was observed in the liver and spleen, indicating hepatobiliary excretion. Imaging showed heterogeneity in dose-distributions for both radionuclides across the tumours. Conclusion: The cytotoxic effect of EGFR-targeted AuNPs is greater in cells with higher EGFR expression. Dose-distributions for individual radiolabelled nanoparticles were heterogeneous across tumours. Further strategies are required to improve the uniformity of dose distribution prior to clinical trials.

Keywords: cancer cells, dose distributions, radionuclide therapy, targeted gold nanoparticles

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732 The Evaluation of Adjuvant Effects of CD154 in a Subunit Vaccine against Classical Swine Fever Virus

Authors: Yu-Chieh Chen, Li-Yun Wang, Chi-Chih Chen, Huy Hùng Đào, Ya-Mei Chen, Ming-Chu Cheng, Wen-Bin Chung, Hso-Chi Chaung, Guan-Ming Ke

Abstract:

Many recent researches have demonstrated that CD154, a protein primarily expressed on activated T cell molecules, has potentially acted as a molecular adjuvant to improve the immunogenicity of subunit vaccines against viral infections. Classical swine fever (CSF) affects the swine industry worldwide that is one of the most devastating and highly contagious pig diseases. It is listed by the World Organization for Animal Health (OIE) as an infectious animal disease that must be reported. Although pigs vaccinated with subunit vaccines can be differentially diagnosed from those infected animals, subunit vaccines usually need adjuvants to enhance and elicit immune responses. In this study, CD154 was linked with CSFV E2 sequences and then expressed in CHO cells to produce the fusion protein as E2-CD154. The porcine specific CpG adjuvant was also used in one of the formulations. The specific pathogen-free pigs (SPF) at the age of 4-week-old were randomly separated into four groups, vaccinated with E2-CpG, E2-CD154, E2-CD154-CpG or the commercial Bayovac® CSF-E2 vaccine and boosted two weeks after primary vaccination. The results showed that the percentages of CD4+ and CD4+IL2+ in peripheral blood mononuclear cells (PBMC) in E2-CD154 vaccinated piglets seven days after primary vaccination were gained by 1-5% relative to the control group. In addition, the percentages of CD4+IFNγ+ T cells had slightly edged up 0.1-0.3% compared with the control group. Also, increased E2-specific IFNγ levels had edged up CD4+CD8+ T cells found in E2-CD154 and E2-CD154-CpG groups, particularly in the E2-CD154-CpG group. These results implicate that CD154 may enhance cellular immunity and synergistically act with species-specific CpG adjuvant as a dual-phase adjuvant. Therefore, the CD154 may be beneficial as a promising adjuvant in subunit vaccines.

Keywords: CD154, CpG adjuvant, cellular immunity, subunit vaccine, pig

Procedia PDF Downloads 56
731 Performance, Yolk and Serum Cholesterol of Shaver-Brown Layers Fed Moringa Leaf Meal and Sun Dried Garlic Powder

Authors: Anselm Onyimonyi, A. Abaponitus

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One hundred and ninety two Shaver-Brown layers aged 40 weeks were used in a 10 weeks feeding trial to investigate the effect of supplementary moringa leaf meal and sun-dried garlic powder (MOGA) on the performance, egg yolk and serum cholesterol profiles of the birds. The birds were randomly assigned to four treatments in a 2 x 2 factorial in a Completely Randomized Design with 48 birds per treatment. Each treatment had 24 replicates with 2 birds, each separately housed in a cell in a battery cage. Birds on treatment 1 received a standard layers mash (16.5% CP and 3000 kcalME/kg) without any MOGA. Treatment 2 birds received the control diet with 5 g moringa leaf meal/kg of feed, treatment 3 received the control diet with 5 g sun-dried garlic powder/kg of feed, treatment 4 had a combination of 5 g each of moringa leaf meal and sun dried garlic powder/kg of feed. Data were kept on daily egg production, egg weight and feed intake. 10 eggs were collected per treatment at the end of the study for yolk cholesterol determination. Blood samples from four birds per treatment were collected and used for the serum cholesterol and triglycerides determination. Results showed that bird on treatment 3 (5% moringa leaf meal/kg of feed) had significantly higher (P < 0.05) Hen Day Egg Production record of 83.3% as against 78.75%, 65.05% and 66.67% recorded for the control, T2 and T4 birds, respectively. Egg weight of 56.39 g recorded for the same birds on treatment 3 was significantly (P< 0.05) lower than the values of 62.61 g, 60.99 g and 59.33 g recorded for birds on T4, T1 and T2, respectively. Yolk and serum cholesterol profiles of the moringa leaf meal fed birds were significantly (P<0.05) lowered when compared to those of the other treatments. Comparatively, the birds on the MOGA diets had significantly reduced yolk and serum cholesterol than the control. It is concluded that supplementation of moringa leaf meal and sun dried garlic powder at the levels used in this study will result in the production of nutritionally healthier eggs with less yolk and serum cholesterol.

Keywords: performance, cholesterol, moringa, garlic

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730 Dwindling the Stability of DNA Sequence by Base Substitution at Intersection of COMT and MIR4761 Gene

Authors: Srishty Gulati, Anju Singh, Shrikant Kukreti

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The manifestation of structural polymorphism in DNA depends on the sequence and surrounding environment. Ample of folded DNA structures have been found in the cellular system out of which DNA hairpins are very common, however, are indispensable due to their role in the replication initiation sites, recombination, transcription regulation, and protein recognition. We enumerate this approach in our study, where the two base substitutions and change in temperature embark destabilization of DNA structure and misbalance the equilibrium between two structures of a sequence present at the overlapping region of the human COMT gene and MIR4761 gene. COMT and MIR4761 gene encodes for catechol-O-methyltransferase (COMT) enzyme and microRNAs (miRNAs), respectively. Environmental changes and errors during cell division lead to genetic abnormalities. The COMT gene entailed in dopamine regulation fosters neurological diseases like Parkinson's disease, schizophrenia, velocardiofacial syndrome, etc. A 19-mer deoxyoligonucleotide sequence 5'-AGGACAAGGTGTGCATGCC-3' (COMT19) is located at exon-4 on chromosome 22 and band q11.2 at the intersection of COMT and MIR4761 gene. Bioinformatics studies suggest that this sequence is conserved in humans and few other organisms and is involved in recognition of transcription factors in the vicinity of 3'-end. Non-denaturating gel electrophoresis and CD spectroscopy of COMT sequences indicate the formation of hairpin type DNA structures. Temperature-dependent CD studies revealed an unusual shift in the slipped DNA-Hairpin DNA equilibrium with the change in temperature. Also, UV-thermal melting techniques suggest that the two base substitutions on the complementary strand of COMT19 did not affect the structure but reduces the stability of duplex. This study gives insight about the possibility of existing structurally polymorphic transient states within DNA segments present at the intersection of COMT and MIR4761 gene.

Keywords: base-substitution, catechol-o-methyltransferase (COMT), hairpin-DNA, structural polymorphism

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729 Optimization and Kinetic Analysis of the Enzymatic Hydrolysis of Oil Palm Empty Fruit Bunch To Xylose Using Crude Xylanase from Trichoderma Viride ITB CC L.67

Authors: Efri Mardawati, Ronny Purwadi, Made Tri Ari Penia Kresnowati, Tjandra Setiadi

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EFB are mainly composed of cellulose (≈ 43%), hemicellulose (≈ 23%) and lignin (≈20%). The palm oil empty fruit bunches (EFB) is the lignosellulosic waste from crude palm oil industries mainly compose of (≈ 43%), hemicellulose (≈ 23%) and lignin (≈20%). Xylan, a polymer made of pentose sugar xylose and the most abundant component of hemicellulose in plant cell wall. Further xylose can be used as a raw material for production of a wide variety of chemicals such as xylitol, which is extensively used in food, pharmaceutical and thin coating applications. Currently, xylose is mostly produced from xylan via chemical hydrolysis processes. However, these processes are normally conducted at a high temperature and pressure, which is costly, and the required downstream processes are relatively complex. As an alternative method, enzymatic hydrolysis of xylan to xylose offers an environmentally friendly biotechnological process, which is performed at ambient temperature and pressure with high specificity and at low cost. This process is catalysed by xylanolytic enzymes that can be produced by some fungal species such as Aspergillus niger, Penicillium crysogenum, Tricoderma reseei, etc. Fungal that will be used to produce crude xylanase enzyme in this study is T. Viride ITB CC L.67. It is the purposes of this research to study the influence of pretreatment of EFB for the enzymatic hydrolysis process, optimation of temperature and pH of the hydrolysis process, the influence of substrate and enzyme concentration to the enzymatic hydrolysis process, the dynamics of hydrolysis process and followingly to study the kinetics of this process. Xylose as the product of enzymatic hydrolysis process analyzed by HPLC. The results show that the thermal pretreatment of EFB enhance the enzymatic hydrolysis process. The enzymatic hydrolysis can be well approached by the Michaelis Menten kinetic model, and kinetic parameters are obtained from experimental data.

Keywords: oil palm empty fruit bunches (EFB), xylose, enzymatic hydrolysis, kinetic modelling

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728 Effect of Engineered Low Glycemic Foods on Cancer Progression and Healthy State

Authors: C. Panebianco, K. Adamberg, S. Adamberg, C. Saracino, M. Jaagura, K. Kolk, A. Di Chio, P. Graziano, R. Vilu, V. Pazienza

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Background/Aims: Despite recent advances in treatment options, a modest impact on the outcome of the pancreatic cancer (PC) is observed so far. Short-term fasting cycles have the potential to improve the efficacy of chemotherapy against PC. However, diseased people may refuse to follow the fasting regimen and fasting may worsen the weight loss often occurring in cancer patients. Therefore, alternative approaches are needed. The aim of this study was to assess the effect of Engineered Low glycemic food ELGIF mimicking diet on growth of cancer cell lines in vitro and in an in vivo pancreatic cancer mouse xenograft model. Materials and Methods: BxPC-3, MiaPaca-2 and Panc-1 cells were cultured in control and ELGIF mimicking diet culturing condition to evaluate the tumor growth and proliferation pathways. Pancreatic cancer xenograft mice were subjected to ELGIF to assess the tumor volume and weight as compared to mice fed with control diet. Results: Pancreatic cancer cells cultured in ELGIF mimicking medium showed decreased levels of proliferation as compared to those cultured in the standard medium. Consistently, xenograft pancreatic cancer mice subjected to ELGIF diet displayed a significant decrease in tumor growth. Conclusion: A positive effect of ELGIF diet on proliferation in vitro is associated with the decrease of tumor progression in the in vivo PC xenograft mouse model. These results suggest that engineered dietary interventions could be supportive as synergistic approach to enhance the efficacy of existing cancer treatments in pancreatic cancer patients.

Keywords: functional food, microbiota, mouse model, pancreatic cancer

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727 Genetic and Virulence Diversity among Alternaria carthami Isolates of India

Authors: Garima Anand, Rupam Kapoor

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Alternaria leaf spot caused by Alternaria carthami is one of the most devastating diseases of safflower. It has resulted in huge losses in crop production and cultivation leading to a fall out of India’s rank as the leading producer of safflower in the world. Understanding the diversity of any pathogen is essential for its management and for the development of disease control strategies. The diversity of A. carthami was therefore analysed on the basis of biochemical, pathogenicity and genetic lines using ISSR markers. Collections and isolations of 95 isolates of A. carthami were made from major safflower producing states of India. Virulence was analysed to evaluate the pathogenic potential of these isolates. The isolates from Bijapur, Dharwad districts (Karnataka), and Parbhani and Solapur districts (Maharashtra) were found to be highly virulent. The virulence assays showed low virulence levels (42%) for the largest part of the population. Biochemical characterization to assess aggressiveness of these isolates was done by estimating the activity of cell wall degrading enzymes where isolates from districts Dharwad, Bijapur of Karnataka and districts Parbhani and Latur of Maharashtra were found to be most aggressive. Genetic diversity among isolates of A. carthami was determined using eighteen ISSR markers. Distance analysis using neighbour joining method and PCoA analysis of the ISSR profiles divided the isolates into three sub-populations. The most virulent isolates clustered in one group in the dendrogram. The study provided no evidence for geographical clustering indicating that isolates are randomly spread across the states, signifying the high potential of the fungus to adapt to diverse regions. The study can, therefore, aid in the breeding and deployment of A. carthami resistant safflower varieties and in the management of Alternaria leaf spot disease.

Keywords: alternaria leaf spot, genetic diversity, pathogenic potential, virulence

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726 Topology Enhancement of a Straight Fin Using a Porous Media Computational Fluid Dynamics Simulation Approach

Authors: S. Wakim, M. Nemer, B. Zeghondy, B. Ghannam, C. Bouallou

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Designing the optimal heat exchanger is still an essential objective to be achieved. Parametrical optimization involves the evaluation of the heat exchanger dimensions to find those that best satisfy certain objectives. This method contributes to an enhanced design rather than an optimized one. On the contrary, topology optimization finds the optimal structure that satisfies the design objectives. The huge development in metal additive manufacturing allowed topology optimization to find its way into engineering applications especially in the aerospace field to optimize metal structures. Using topology optimization in 3d heat and mass transfer problems requires huge computational time, therefore coupling it with CFD simulations can reduce this it. However, existed CFD models cannot be coupled with topology optimization. The CFD model must allow creating a uniform mesh despite the initial geometry complexity and also to swap the cells from fluid to solid and vice versa. In this paper, a porous media approach compatible with topology optimization criteria is developed. It consists of modeling the fluid region of the heat exchanger as porous media having high porosity and similarly the solid region is modeled as porous media having low porosity. The switching from fluid to solid cells required by topology optimization is simply done by changing each cell porosity using a user defined function. This model is tested on a plate and fin heat exchanger and validated by comparing its results to experimental data and simulations results. Furthermore, this model is used to perform a material reallocation based on local criteria to optimize a plate and fin heat exchanger under a constant heat duty constraint. The optimized fin uses 20% fewer materials than the first while the pressure drop is reduced by about 13%.

Keywords: computational methods, finite element method, heat exchanger, porous media, topology optimization

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725 Bulk Modification of Poly(Dimethylsiloxane) for Biomedical Applications

Authors: A. Aslihan Gokaltun, Martin L. Yarmush, Ayse Asatekin, O. Berk Usta

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In the last decade microfabrication processes including rapid prototyping techniques have advanced rapidly and achieved a fairly matured stage. These advances encouraged and enabled the use of microfluidic devices by a wider range of users with applications in biological separations, and cell and organoid cultures. Accordingly, a significant current challenge in the field is controlling biomolecular interactions at interfaces and the development of novel biomaterials to satisfy the unique needs of the biomedical applications. Poly(dimethylsiloxane) (PDMS) is by far the most preferred material in the fabrication of microfluidic devices. This can be attributed its favorable properties, including: (1) simple fabrication by replica molding, (2) good mechanical properties, (3) excellent optical transparency from 240 to 1100 nm, (4) biocompatibility and non-toxicity, and (5) high gas permeability. However, high hydrophobicity (water contact angle ~108°±7°) of PDMS often limits its applications where solutions containing biological samples are concerned. In our study, we created a simple, easy method for modifying the surface chemistry of PDMS microfluidic devices through the addition of surface-segregating additives during manufacture. In this method, a surface segregating copolymer is added to precursors for silicone and the desired device is manufactured following the usual methods. When the device surface is in contact with an aqueous solution, the copolymer self-organizes to expose its hydrophilic segments to the surface, making the surface of the silicone device more hydrophilic. This can lead to several improved performance criteria including lower fouling, lower non-specific adsorption, and better wettability. Specifically, this approach is expected to be useful for the manufacture of microfluidic devices. It is also likely to be useful for manufacturing silicone tubing and other materials, biomaterial applications, and surface coatings.

Keywords: microfluidics, non-specific protein adsorption, PDMS, PEG, copolymer

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724 Optimization of Platinum Utilization by Using Stochastic Modeling of Carbon-Supported Platinum Catalyst Layer of Proton Exchange Membrane Fuel Cells

Authors: Ali Akbar, Seungho Shin, Sukkee Um

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The composition of catalyst layers (CLs) plays an important role in the overall performance and cost of the proton exchange membrane fuel cells (PEMFCs). Low platinum loading, high utilization, and more durable catalyst still remain as critical challenges for PEMFCs. In this study, a three-dimensional material network model is developed to visualize the nanostructure of carbon supported platinum Pt/C and Pt/VACNT catalysts in pursuance of maximizing the catalyst utilization. The quadruple-phase randomly generated CLs domain is formulated using quasi-random stochastic Monte Carlo-based method. This unique statistical approach of four-phase (i.e., pore, ionomer, carbon, and platinum) model is closely mimic of manufacturing process of CLs. Various CLs compositions are simulated to elucidate the effect of electrons, ions, and mass transport paths on the catalyst utilization factor. Based on simulation results, the effect of key factors such as porosity, ionomer contents and Pt weight percentage in Pt/C catalyst have been investigated at the represented elementary volume (REV) scale. The results show that the relationship between ionomer content and Pt utilization is in good agreement with existing experimental calculations. Furthermore, this model is implemented on the state-of-the-art Pt/VACNT CLs. The simulation results on Pt/VACNT based CLs show exceptionally high catalyst utilization as compared to Pt/C with different composition ratios. More importantly, this study reveals that the maximum catalyst utilization depends on the distance spacing between the carbon nanotubes for Pt/VACNT. The current simulation results are expected to be utilized in the optimization of nano-structural construction and composition of Pt/C and Pt/VACNT CLs.

Keywords: catalyst layer, platinum utilization, proton exchange membrane fuel cell, stochastic modeling

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723 Study of COVID-19 Intensity Correlated with Specific Biomarkers and Environmental Factors

Authors: Satendra Pal Singh, Dalip Kr. Kakru, Jyoti Mishra, Rajesh Thakur, Tarana Sarwat

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COVID-19 is still an intrigue as far as morbidity or mortality is concerned. The rate of recovery varies from person to person, & it depends upon the accessibility of the healthcare system and the roles played by the physicians and caregivers. It is envisaged that with the passage of time, people would become immune to this virus, and those who are vulnerable would sustain themselves with the help of vaccines. The proposed study deals with the severeness of COVID-19 is associated with some specific biomarkers linked to correlate age and gender. We will be assessing the overall homeostasis of the persons who were affected by the coronavirus infection and also of those who recovered from it. Some people show more severe effects, while others show very mild symptoms, however, they show low CT values. Thus far, it is unclear why the new strain of Covid has different effects on different people in terms of age, gender, and ABO blood typing. According to data, the fatality rate with heart disease was 10.5 percent, 7.3 percent were diabetic, and 6 percent who are already infected from other comorbidities. However, some COVID-19 cases are worse than others & it is not fully explainable as of date. Overall data show that the ABO blood group is effective or prone to the risk of SARS-COV2 infection, while another study also shows the phenotypic effects of the blood group related to covid. It is an accepted fact that females have more strong immune systems than males, which may be related to the fact that females have two ‘X’ chromosomes, which might contain a more effective immunity booster gene on the X chromosome, and are capable to protect the female. Also specific sex hormones also induce a better immune response in a specific gender. This calls for in-depth analysis to be able to gain insight into this dilemma. COVID-19 is still not fully characterized, and thus we are not very familiar with its biology, mode of infection, susceptibility, and overall viral load in the human body. How many virus particles are needed to infect a person? How, then, comorbidity contribute to coronavirus infection? Since the emergence of this virus in 2020, a large number of papers have been published, and seemingly, vaccines have been prepared. But still, a large number of questions remain unanswered. The proneness of humans for infection by covid-19 needs to be established to be able to develop a better strategy to fight this virus. Our study will be on the Impact of demography on the Severity of covid-19 infection & at the same time, will look into gender-specific sensitivity of Covid-19 and the Operational variation of different biochemical markers in Covid-19 positive patients. Besides, we will be studying the co-relation, if any, of COVID severity & ABO Blood group type and the occurrence of the most common blood group type amongst positive patience.

Keywords: coronavirus, ABO blood group, age, gender

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722 Synthesis of a Hybrid of PEG-b-PCL and G1-PEA Dendrimer Based Six-Armed Star Polymer for Nano Delivery of Vancomycin

Authors: Calvin A. Omolo, Rahul S. Kalhapure, Mahantesh Jadhav, Sanjeev Rambharose, Chunderika Mocktar, Thirumala Govender

Abstract:

Treatment of infections is compromised by limitations of conventional dosage forms and drug resistance. Nanocarrier system is a strategy to overcome these challenges and improve therapy. Thus, the development of novel materials for drug delivery via nanocarriers is essential. The aim of the study was to synthesize a multi-arm polymer (6-mPEPEA) for enhanced activity of vancomycin (VM) against susceptible and resistant Staphylococcus aureus (MRSA). The synthesis steps of the star polymer followed reported procedures. The synthesized 6-mPEPEA was characterized by FTIR, ¹H and ¹³CNMR and MTT assays. VM loaded micelles were prepared from 6-mPEPEA and characterized for size, polydispersity index (PI) and surface charge (ZP) (Dynamic Light Scattering), morphology by TEM, drug loading (UV Spectrophotometry), drug release (dialysis bag), in vitro and in vivo efficacy against sensitive and resistant S. aureus. 6-mPEPEA was synthesized, and its structure was confirmed. MTT assays confirmed its nontoxic nature with a high cell viability (77%-85%). Unimolecular spherical micelles were prepared. Size, PI, and ZP was 52.48 ± 2.6 nm, 0.103 ± 0.047, -7.3 ± 1.3 mV, respectively and drug loading was 62.24 ± 3.8%. There was a 91% drug release from VCM-6-mPEPEA after 72 hours. In vitro antibacterial test revealed that VM-6-mPEPEA had 8 and 16-fold greater activity against S. aureus and MRSA when compared to bare VM. Further investigations using flow cytometry showed that VM-6-mPEPEA had 99.5% killing rate of MRSA at the MIC concentration. In vivo antibacterial activity revealed that treatment with VM-6-mPEPEA had a 190 and a 15-fold reduction in the MRSA load in untreated and VM treated respectively. These findings confirmed the potential of 6-mPEPEA as a promising bio-degradable nanocarrier for antibiotic delivery to improve treatment of bacterial infections.

Keywords: biosafe, MRSA, nanocarrier, resistance, unimolecular-micelles

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721 SUMOylation Enhances Nurr1/1a Mediated Transactivation in a Neuronal Cell Type

Authors: Jade Edey, Andrew Bennett, Gareth Hathway

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Nuclear receptor-related 1 protein (also known as Nurr1 or NR4A2) is an orphan nuclear receptor which plays a vital role in the development, survival and maintenance of dopaminergic (DA) neurons particularly in the substantia nigra (SN). Increasing research has investigated Nurr1’s additional role within microglia and astrocytes where it has been suggested to act as a negative regulator of inflammation; potentially offering neuroprotection. Considering both DA neurodegeneration and neuroinflammation are commonly accepted constituents of Parkinson’s Disease (PD), understanding the mechanisms by which Nurr1 regulates inflammatory processes could provide an attractive therapeutic target. Nurr1 regulates inflammation via a transrepressive mechanism possibly dependent upon SUMOylation. In addition, Nurr1 can transactivate numerous genes involved in DA synthesis, such as Tyrosine Hydroxylase (TH). A C-terminal splice variant of Nurr1, Nurr-1a, has been reported in both neuronal and glial cells. However, research into its transcriptional activity is minimal. We employed in vitro methods such as SUMO-Pulldown experiments alongside Luciferase reporter assays to investigate the SUMOylation status and transactivation capabilities of Nurr1 and Nurr-1a respectively. The SUMO-Pulldown assay demonstrated Nurr-1a undergoes significantly more SUMO modification than its full-length variant. Consequently, despite having less transcriptional activation than Nurr1, Nurr1a may play a more prominent role in repression of microglial inflammation. Contrary to published literature we also identified that SUMOylation enhances transcriptional activation by Nurr1 and Nurr1a. SUMOylation-dependent increases in Nurr1 and Nurr1a transcriptional activation were only evident in neuronal SHSY5Y cells but not in HEK293 cells. This research provides novel insight into the regulation of Nurr-1a and indicates differential effects of SUMOylation dependent regulation in neuronal and inflammatory cells.

Keywords: nuclear receptors, Parkinson’s disease, inflammation, transcriptional regulation

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720 Genetic Association and Functional Significance of Matrix Metalloproteinase-14 Promoter Variants rs1004030 and rs1003349 in Gallbladder Cancer Pathogenesis

Authors: J. Vinay , Kusumbati Besra, Niharika Pattnaik, Shivaram Prasad Singh, Manjusha Dixit

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Gallbladder cancer (GBC) is rare but highly malignant cancer; its prevalence is more in certain geographical regions and ethnic groups, which include the Northern and Eastern states of India. Previous studies in India have reported genetic predisposition as one of the risk factors in GBC pathogenesis. Although the matrix metalloproteinase-14 (MMP14) is a well-known modulator of the tumor microenvironment and tumorigenesis and TCGA data also suggests its upregulation yet, its role in the genetic predisposition for GBC is completely unknown. We elucidated the role of MMP14 promoter variants as genetic risk factors and their implications in expression modulation. We screened MMP14 promoter variants association with GBC using Sanger’s sequencing in approximately 300 GBC and 300 control subjects and 26 GBC tissue samples of Indian ethnicity. The immunohistochemistry was used to check the MMP14 protein expression in GBC tissue samples. The role of promoter variants on expression levels was elucidated using a luciferase reporter assay. The variants rs1004030 (p-value = 0.0001) and rs1003349 (p-value = 0.0008) were significantly associated with gallbladder cancer. The luciferase assay in two different cell lines, HEK-293 (p = 0.0006) and TGBC1TKB (p = 0.0036) showed a significant increase in relative luciferase activity in the presence of risk alleles for both the single nucleotide polymorphisms (SNPs). Similarly, genotype-phenotype correlation in patients samples confirmed that the presence of risk alleles at rs1004030 and rs1003349 increased MMP14 expression. Overall, this study unravels the genetic association of MMP14 promoter variants with gallbladder cancer, which may contribute to pathogenesis by increasing its expression.

Keywords: gallbladder cancer, matrix metalloproteinase-14, single nucleotide polymorphism, case control study, genetic association study

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719 Hyponatremia in Community-Acquired Pneumonia

Authors: Emna Ketata, Wafa Farhat

Abstract:

Introduction: Hyponatremia is defined by a blood sodium level of ≤ 136 mmol/L; it is associated with a high risk of morbidity and mortality in the emergency room. This was explained by transit disorders, including diarrhea and inappropriate antidiuretic hormone secretion (Syndrome of inappropriate antidiuretic hormone secretion). Pneumonia can cause dyspnea, stress-causing SIADH and digestive symptoms (diarrhea and vomiting). Aim: The purpose of this study was to determine the link between pneumonia and hyponatremia as a predictor of patient’s prognosis and intra-hospital mortality. Methodology: This is a prospective observational study over a period of 3 years in the emergency department. Inclusion :patients (age > 14 years), with clinical signs in favor of pneumonia. Natremia was measured. Natremia was classified as mild to moderate with a blood sodium level between 121 and 135 mmol/L and as severe with a blood sodium level ≤ 120 mmol/L. Results: This study showed an average serum sodium value of 135 mmol/L (range 114–159 mmol/L) in these patients. Hyponatremia was observed in 123 patients (43.6%), 115 patients (97,8%) had mild to moderate hyponatremia and 2,8% had severe hyponatremia. The mean age was 65±17 years with a sex ratio of 1.05. The main reason for consultation in patients with hyponatremia was cough in 58 patients (47.2%), and digestive symptoms were present in 25 patients (20.3. An altered state of consciousness was observed in 11 patients (3%). Patients with hyponatremia had greater heart rate (p=0.02),white blood cell count (p=0.009) , plasmatic lactate (p=0.002) and higher rate of pneumonia recurrence (p=0.001) .In addition, 80% of them have a positive CURB65 score (>=2). hyponatremia had higher rates of use of oxygen therapy compared to patients with normo-natremia (54% vs. 45%). The analytical study showed that hyponatremia is significantly associated with intra-hospital mortality with( p=0.01), severe hyponatremia p=0.04. Conclusion: Hyponatremia is a predictor of mortality and worse prognosis. Recognition of the pathophysiological mechanisms of hyponatremia in pneumonia will probably allow better management of it.

Keywords: oxygenotherapy, mortality, recurrence, positif curb65

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718 Analyzing the Relationship between Physical Fitness and Academic Achievement in Chinese High School Students

Authors: Juan Li, Hui Tian, Min Wang

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In China, under the considerable pressure of 'Gaokao' –the highly competitive college entrance examination, high school teachers and parents often worry that doing physical activity would take away the students’ precious study time and may have a negative impact on the academic grades. There was a tendency to achieve high academic scores at the cost of physical exercise. Therefore, the purpose of this study was to examine the relationship between the physical fitness and academic achievement of Chinese high school students. The participants were 968 grade one (N=457) and grade two students (N=511) with an average age of 16 years from three high schools of different levels in Beijing, China. 479 were boys, and 489 were girls. One of the schools is a top high school in China, another is a key high school in Beijing, and the other is an ordinary high school. All analyses were weighted using SAS 9.4 to ensure the representatives of the sample. The weights were based on 12 strata of schools, sex, and grades. Physical fitness data were collected using the scores of the National Physical Fitness Test, which is an annual official test administered by the Ministry of Education in China. It includes 50m run, sits and reach test, standing long jump, 1000m run (for boys), 800m run (for girls), pull-ups for 1 minute (for boys), and bent-knee sit-ups for 1 minute (for girls). The test is an overall evaluation of the students’ physical health on the major indexes of strength, endurance, flexibility, and cardiorespiratory function. Academic scores were obtained from the three schools with the students’ consent. The statistical analysis was conducted with SPSS 24. Independent-Samples T-test was used to examine the gender group differences. Spearman’s Rho bivariate correlation was adopted to test for associations between physical test results and academic performance. Statistical significance was set at p<.05. The study found that girls obtained higher fitness scores than boys (p=.000). The girls’ physical fitness test scores were positively associated with the total academic grades (rs=.103, p=.029), English (rs=.096, p=.042), physics (rs=.202, p=.000) and chemistry scores (rs=.131, p=.009). No significant relationship was observed in boys. Cardiorespiratory fitness had a positive association with physics (rs=.196, p=.000) and biology scores (rs=.168, p=.023) in girls, and with English score in boys (rs=.104, p=.029). A possible explanation for the greater association between physical fitness and academic achievement in girls rather than boys was that girls showed stronger motivation in achieving high scores in whether academic tests or fitness tests. More driven by the test results, girls probably tended to invest more time and energy in training for the fitness test. Higher fitness levels were associated with an academic benefit among girls generally in Chinese high schools. Therefore, physical fitness needs to be given greater emphasis among Chinese adolescents and gender differences need to be taken into consideration.

Keywords: physical fitness; adolescents; academic achievement; high school

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717 Anti-Parasite Targeting with Amino Acid-Capped Nanoparticles Modulates Multiple Cellular Processes in Host

Authors: Oluyomi Stephen Adeyemi, Kentaro Kato

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Toxoplasma gondii is the etiological agent of toxoplasmosis, a common parasitic disease capable of infecting a range of hosts, including nearly one-third of the human population. Current treatment options for toxoplasmosis patients are limited. In consequence, toxoplasmosis represents a large global burden that is further enhanced by the shortcomings of the current therapeutic options. These factors underscore the need for better anti-T. gondii agents and/or new treatment approach. In the present study, we sought to find out whether preparing and capping nanoparticles (NPs) in amino acids, would enhance specificity toward the parasite versus the host cell. The selection of amino acids was premised on the fact that T. gondii is auxotrophic for some amino acids. The amino acid-nanoparticles (amino-NPs) were synthesized, purified and characterized following established protocols. Next, we tested to determine the anti-T. gondii activity of the amino-NPs using in vitro experimental model of infection. Overall, our data show evidence that supports enhanced and excellent selective action against the parasite versus the host cells by amino-NPs. The findings are promising and provide additional support that warrants exploring the prospects of NPs as alternative anti-parasite agents. In addition, the anti-parasite action by amino-NPs indicates that nutritional requirement of parasite may represent a viable target in the development of better alternative anti-parasite agents. Furthermore, data suggest the anti-parasite mechanism of the amino-NPs involves multiple cellular processes including the production of reactive oxygen species (ROS), modulation of hypoxia-inducing factor-1 alpha (HIF-1α) as well as the activation of kynurenine pathway. Taken together, findings highlight further, the prospects of NPs as alternative source of anti-parasite agents.

Keywords: drug discovery, infectious diseases, mode of action, nanomedicine

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716 Aspirin Loaded Poly-L-Lactic Acid Nanofibers and Their Potentials as Small Diameter Vascular Grafts

Authors: Mahboubeh Kabiri, Saba Aslani

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Among various approaches used for the treatment of cardiovascular diseases, the occlusion of the small-diameter vascular graft (SDVG) is still an unresolved problem which seeks further research to address them. Though autografts are now the gold standards to be replaced for blocked coronary arteries, they suffer from inadequate quality and quantity. On the other hand, the major problems of the tissue engineered grafts are thrombosis and intimal hyperplasia. Provision of a suitable spatiotemporal release pattern of anticoagulant agents such as heparin and aspirin can be a step forward to overcome such issues . Herein, we fabricated electrospun scaffolds from FDA (Food and Drug Administration) approved poly-L-lactic acid (PLLA) with aspirin loaded into the nanofibers. Also, we surface coated the scaffolds with Amniotic Membrane lysate as a source for natural elastic polymers and a mimic of endothelial basement membrane. The scaffolds were characterized thoroughly structurally and mechanically for their morphology, fiber orientation, tensile strength, hydrophilicity, cytotoxicity, aspirin release and cell attachment support. According to the scanning electron microscopy (SEM) images, the size of fibers ranged from 250 to 500 nm. The scaffolds showed appropriate tensile strength expected for vascular grafts. Cellular attachment, growth, and infiltration were proved using SEM and MTT (3-(4,5-Dimethylthiazol-2-Yl)-2,5-Diphenyltetrazolium Bromide) assay. Drug-loaded scaffolds showed a sustained release profile of aspirin in 7 days. An enhanced cytocompatibility was observed in AM-coated electrospun PLLA fibers compared to uncoated scaffolds. Our results together indicated that AM lysate coated ASA releasing scaffolds have promising potentials for development of a biocompatible SDVG.

Keywords: vascular tissue engineering, vascular grafts, anticoagulant agent, aspirin, amniotic membrane

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715 Development of Monoclonal Antibodies against the Acute Hepatopancreatic Necrosis Disease Toxins

Authors: Naveen Kumar B. T., Anuj Tyagi, Niraj Kumar Singh, Visanu Boonyawiwat, Shanthanagouda A. H., Orawan Boodde, Shankar K. M., Prakash Patil, Shubhkaramjeet Kaur

Abstract:

Since 2009, Acute Hepatopancreatic Necrosis Disease (AHPND) outbreaks have increased rapidly, and these have led to the major economic losses to the global shrimp industry. In comparison to other treatments, passive immunity and monoclonal antibody (MAb) based farmer level kit have proved their importance in controlling and treating the diseases in the shrimp industry. In the present study, MAbs were produced against the recombinant PirB protein Vibrio parahaemolyticus strain causing AHPND. Briefly, Balb/C mice were immunized with rPirB at 15 days interval, and antibody titer was determined by ELISA. Spleen cells from mice showing high antibody titer were fused with SP2O myeloma cells for hybridoma production. Among 130 hybridomas, four showed high antibody titer and positive reactivity in an immunoblot assay. In Western blot assay, three out of four MAbs (4C4, 2C2 and 4G3) showed reactivity to rPirB protein. However, in the natural host, only Mab clone 4G3 show strong reactivity (with a strain of V. parahemolyticus causing EMS/AHPND). These clones also showed reactivity with less than 20 kDa proteins in AHPND free V. parahaemolyticus (Thailand stain). Further, on from MAb 4G3 clone, four panels of single cell MAbs clones (G3F5, G3B8, G3H2, and G3D6) were produced of which three showed strong positive reactivity to rPirB protein in the Western blot. These MAbs have potential for controlling and prevention of the AHPND through passive immunity and development of filed level rapid diagnostic kits.

Keywords: shrimp, economic loss, AHPND, MAb

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714 Astronomy in the Education Area: A Narrative Review

Authors: Isabella Lima Leite de Freitas

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The importance of astronomy for humanity is unquestionable. Despite being a robust science, capable of bringing new discoveries every day and quickly increasing the ability of researchers to understand the universe more deeply, scientific research in this area can also help in various applications outside the domain of astronomy. The objective of this study was to review and conduct a descriptive analysis of published studies that presented the importance of astronomy in the area of education. A narrative review of the literature has been performed, considering the articles published in the last five years. As astronomy involves the study of physics, chemistry, biology, mathematics and technology, one of the studies evaluated presented astronomy as the gateway to science, demonstrating the presence of astronomy in 52 school curricula in 37 countries, with celestial movement the dominant content area. Another intervention study, evaluating individuals aged 4-5 years, demonstrated that the attribution of personal characteristics to cosmic bodies, in addition to the use of comprehensive astronomy concepts, favored the learning of science in preschool-age children, considering the use of practical activities of accompaniment and free drawing. Aiming to measure scientific literacy, another study developed in Turkey, motivated the authorities of this country to change the teaching materials and curriculum of secondary schools after the term “astronomy” appeared as one of the most attractive subjects for young people aged 15 to 24. There are also reports in the literature of the use of pedagogical tools, such as the representation of the Solar System on a human scale, where students can walk along the orbits of the planets while studying the laws of dynamics. The use of this tool favored the teaching of the relationship between distance, duration and speed over the period of the planets, in addition to improving the motivation and well-being of students aged 14-16. An important impact of astronomy on education was demonstrated in the study that evaluated the participation of high school students in the Astronomical Olympiads and the International Astronomy Olympiad. The study concluded that these Olympics have considerable influence on students who pursue a career in teaching or research later on, many of whom are in the area of astronomy itself. In addition, the literature indicates that the teaching of astronomy in the digital age has facilitated the availability of data for researchers, but also for the general population. This fact can increase even more the curiosity that the astronomy area has always instilled in people and promote the dissemination of knowledge on an expanded scale. Currently, astronomy has been considered an important ally in strengthening the school curricula of children, adolescents and young adults. This has been used as teaching tools, in addition to being extremely useful for scientific literacy, being increasingly used in the area of education.

Keywords: astronomy, education area, teaching, review

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713 Sudanese Dietitian’s Role in the Provision of Parenteral Nutrition: The Past, Present, and Future

Authors: Reem Osama Yousif Ali, Osama Yousif Ali Al Gibali

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Introduction: Balanced nutrition is undeniably essential for maintaining health, body functions, and integrity of cell metabolism; however, some sick patients cannot tolerate oral or enteral feeding to meet their nutritional needs, so partial or total parenteral nutrition (PN) may be the most suitable alternative route in such situations. Dietitians are fundamental personnel among the medical team to ensure the proper provision of PN service, which was introduced in Sudan in the 1980s. Objective: The study aimed to recognize the dietitians' awareness of parenteral nutrition and their role in providing this service in Sudan – Khartoum State. Methodology: Formulated questionnaire forms composed of twelve questions were distributed to the dietitians working in four tertiary level hospitals. Results: The majority (75%) of the responded dietitians had reasonable knowledge about the importance of PN, its advantages, and its indications. Sixty percent of them were mindful of the PN side effects. Most of the dietitians were aware of the different assessment measurements and PN calculations and were exposed in their clinical practice to patients who were in need of PN, but only a few of them (about 30%) had the actual chance to participate in the formulation and application of PN therapy. The unavailability of the multidisciplinary team, lack of the required equipment and financial support, and associated complications were basic obstacles to the provision of long-term PN service in Khartoum state hospitals. Conclusion: Although dietitians in Khartoum state hospitals have good information about PN definition, indications, accesses, and assessment measures, they do not have enough knowledge and clinical exposure that make them confident to provide the PN service. Establishing a few models of parenteral nutrition units in tertiary hospitals will be of great help, as well as providing the dietitian's training in the area of parenteral nutrition. Further study can explore more requirements to run this service.

Keywords: nutrition support, dietitian, Sudan, parenteral nutrition, nutrition support team

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712 DNA Methylation 6mA and Histone Methylation Involved in Multi-/Trans-Generational Reproductive Effects in Caenorhabditis elegans Induced by Atrazine

Authors: Jiechen Yin, Xiang Hong, Ran Liu

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Atrazine (ATR), a widely used triazine herbicide, is an environmental endocrine disruptor that can cause health problems. However, whether there are multi/trans-generational reproductive impacts of ATR have not been studied to our best knowledge. Therefore, in this study, Caenorhabditis elegans was used as a preferable model organism to identify the multi/trans-generational reproductive toxicity of ATR. L1 larvae were exposed to different concentrations (0.0004–40 mg/L) of ATR for 48 h. Successive generations (F1 to F5) were fed without ATR and consecutive exposure. The results showed that ATR exposure during P0 decreased fecundity, including a reduction in fertilized eggs, oocytes, and ovulation rate, delayed gonadal development, and decreased the relative area of the gonad arm and germ cell number. Furthermore, continuous ATR exposure (P0–F5) causes a significant increase in reproductive toxicity in subsequent generations, although no significant toxicity occurred in the P0 generation after exposure to environmental-related concentrations, suggesting that ATR exposure might have cumulative effects. Likewise, parental exposure to ATR caused transgenerational toxicity impairments. Interestingly, reproductive toxicity not development toxicity was transmitted to several generations (F1–F4), and the F2 generation showed the most notable changes. QRT-PCR results showed that genes related to DNA methylation 6mA (damt-1, nmad-1) and histone H3 methylation (mes-4, met-2, set-25, set-2, and utx-1) can also be passed on to offspring. The function of H3K4 and H3K9 methylation were explored by using loss-of-function mutants for set-2, set-25, and met-2. Transmissible reproductive toxicity was absent in met-2(n4256), set-2(ok952), and set-25(n5021) mutants, which suggests that the histone methyltransferases H3K4 and H3K9 activity are indispensable for the transgenerational effect of ATR. Finally, the downstream genes of DNA methylation and histone H3 methylation were determined. ATR upregulated the expression of ZC317.7, hsp-6, and hsp-60. Mitochondrial stress in parental generation dependent transcription 6mA modifiers may establish these epigenetic marks in progeny.

Keywords: ATR, Caenorhabditis elegans, multi-/trans-generation, reproductive toxicity

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711 Soluble CD36 and Cardiovascular Risk in Middle-Aged Subjects

Authors: Mohammad Alkhatatbeh, Nehad Ayoub, Nizar Mhaidat, Nesreen Saadeh, Lisa Lincz

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CD36 is involved in the development of atherosclerosis by enhancing macrophage endocytosis of oxidized-low density lipoproteins and foam cell formation. Soluble CD36 (sCD36) was found to be elevated in type 2 diabetic patients and was supposed to act as a marker of insulin resistance and atherosclerosis. In young subjects, sCD36 was associated with cardiovascular risk factors including obesity and hypertriglyceridemia. This study was conducted to further investigate the relationship between plasma sCD36 and cardiovascular risk factors among middle-aged patients with metabolic syndrome (MetS) and healthy controls. SCD36 concentrations were determined by enzyme-linked immunosorbent assays (ELISA) for 41 patients with MetS and 36 healthy controls. Data for other variables were obtained from patients' medical records. SCD36 concentrations were relatively low compared to most other studies and were not significantly different between the MetS group and controls (P-value=0.17). SCD36 was also not correlated with age, body mass index, glucose, lipid profile, serum electrolytes and blood counts. SCD36 was not significantly different between subjects with obesity, hyperglycemia, dyslipidemia, hypertension or cardiovascular disease and those without these abnormalities (P-value > 0.05). The inconsistency between results reported in this study and other studies may be unique to the study population or be a result of the lack of a reliable standardized method for determining absolute sCD36 concentrations. However, further investigations are required to assess CD36 tissue expression in the study population and to assess the accuracy of various commercially available sCD36 ELISA kits. Thus, the availability of a standardized simple sCD36 ELISA that could be performed in any basic laboratory would be more favorable to the specialized flow cytometry methods that detect CD36+ microparticles if it was to be used as a biomarker.

Keywords: metabolic syndrome, CD36, cardiovascular risk, obesity, type 2 diabetes mellitus

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710 Epigenetic and Archeology: A Quest to Re-Read Humanity

Authors: Salma A. Mahmoud

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Epigenetic, or alteration in gene expression influenced by extragenetic factors, has emerged as one of the most promising areas that will address some of the gaps in our current knowledge in understanding patterns of human variation. In the last decade, the research investigating epigenetic mechanisms in many fields has flourished and witnessed significant progress. It paved the way for a new era of integrated research especially between anthropology/archeology and life sciences. Skeletal remains are considered the most significant source of information for studying human variations across history, and by utilizing these valuable remains, we can interpret the past events, cultures and populations. In addition to archeological, historical and anthropological importance, studying bones has great implications in other fields such as medicine and science. Bones also can hold within them the secrets of the future as they can act as predictive tools for health, society characteristics and dietary requirements. Bones in their basic forms are composed of cells (osteocytes) that are affected by both genetic and environmental factors, which can only explain a small part of their variability. The primary objective of this project is to examine the epigenetic landscape/signature within bones of archeological remains as a novel marker that could reveal new ways to conceptualize chronological events, gender differences, social status and ecological variations. We attempted here to address discrepancies in common variants such as methylome as well as novel epigenetic regulators such as chromatin remodelers, which to our best knowledge have not yet been investigated by anthropologists/ paleoepigenetists using plethora of techniques (biological, computational, and statistical). Moreover, extracting epigenetic information from bones will highlight the importance of osseous material as a vector to study human beings in several contexts (social, cultural and environmental), and strengthen their essential role as model systems that can be used to investigate and construct various cultural, political and economic events. We also address all steps required to plan and conduct an epigenetic analysis from bone materials (modern and ancient) as well as discussing the key challenges facing researchers aiming to investigate this field. In conclusion, this project will serve as a primer for bioarcheologists/anthropologists and human biologists interested in incorporating epigenetic data into their research programs. Understanding the roles of epigenetic mechanisms in bone structure and function will be very helpful for a better comprehension of their biology and highlighting their essentiality as interdisciplinary vectors and a key material in archeological research.

Keywords: epigenetics, archeology, bones, chromatin, methylome

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709 Effect of Smoking on Tear Break-Up Time and Basal Tear Secretion

Authors: Kalsoom Rani

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Tobacco contains nicotine, which causes addiction to many toxic chemicals. In the world, people consume it in the form of smoke, chew, and sniffing, smoke of it is composed of almost 7000 active chemicals, which are very harmful to human health as well as for eye health, inhalation of tobacco smoke and fumes can accelerate and cause many blinding eye diseases. Dry eye and smoking have not been covered extensively in researches; more studies are required to unveil the relationship between smoking and dry eye. This study was conducted to determine the quantity and quality of tears in smokers. 60 subjects participated in the study, which was divided into two groups on the basis of consumption of cigarettes per day with age matched non smokers of 15-50 years. All participants have gone through a study based questioner, eye examination, and diagnostic 'Dry Eye Tests' for evaporative tears evaluation and measurement of basal tear secretion. Subjects were included in the criteria of 10 cigarettes per day with a minimum duration of 1 year; passive smokers for control groups were excluded. The study was carried out in a Medina Teaching Hospital, Faisalabad, Pakistan, ophthalmology department for the duration of 8 months. Mean values for tear break up time (TBUT), was reported 10sec with SD of +3.74 in controlled group, 5sec with SD + 2.32 in smokers and 4sec SD +3.77 heavy smokers in right eye (RE) and left eye (LE) 10.35sec with SD of +3.88 in controlled 5sec with SD + 2.3 in smokers and much reduced TBUT in heavy smokers was 3.85sec SD+2.20. Smoking has a very strong association with TRUT with a significance of P=.00 both eyes. Mean Schirmer-I value of the subjects was reported 12.6mm with SD + 8.37 in RE and 12.59mm with SD + 8.96 LE. The mean Schirmer-II test value was reported in the right, and left eye with a mean value for control was 20.23mm with SD + 8.93, 20.75mm with SD + 8.84 respectively, and in Smokers 9.90mm with SD + 5.74, and 10.07mm with SD + 6.98, and in heavy smokers 7.7mm, SD + 3.22 and 6.9, SD + 3.50 mm, association with smoking showed p=.001 in RE and .003 in LE. Smoking has deteriorated effect on both evaporative tear and aqueous tear secretion and causing symptoms of dry eye burning, itching, redness, and watering with epithelial cell damage.

Keywords: tear break-up time, basal tear secretion, smokers, dry eye

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708 Occurence And Management Of Coliform Bacteria On Tomatoes

Authors: Cho Achidi

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Tomato is a crucial food crop significantly contributes to global food and nutrition security. However, postharvest losses severely limit its role. Therefore, it is necessary to develop sustainable strategies to minimize these losses and improve the shelf-life of tomato fruits. One of the major concerns is bacterial infections, particularly by faecal coliform bacteria, which can cause food poisoning and illnesses like diarrhoea and dysentery. This study seeks to identify the presence of coliform bacteria on tomato fruits in fields and markets in Muea, Buea Municipality. The study also evaluated different management strategies to reduce the bacterial incidence and load on tomato fruits. A total of 200 fruits were sampled for both the coliform survey and shelf-life analysis. Ten farmers and traders provided samples, including asymptomatic and symptomatic tomato fruits. The samples designated for shelf-life analysis were treated with Aquatab, warm water, lemon, and onion. The results indicated that out of the 80 symptomatic samples collected, 12.5% contained faecal and total coliform species. Among the ten farms sampled, 14% were infected with coliform bacteria, with the highest infestation rate of 60% recorded in field 4. Furthermore, 15% of the asymptomatic tomato fruits were found to be infected by coliform bacteria. Regarding the management strategies, Aquatabs exhibited the highest efficacy in reducing the incidence of coliform bacteria on tomato fruits, followed by onion and lemon extracts. Although hot water treatment effectively removed bacteria from the fruits, damaging the cell wall negatively affected their shelf-life. Overall, this study emphasizes the severity of coliform bacterial pathogens in the Muea area, particularly their occurrence on asymptomatic tomatoes, which poses a significant concern for plant quarantine services. It also demonstrates potential options for mitigating this bacterial challenge.

Keywords: tomato, shelf-life analysis, food and nutrition security, coliform bbacteria

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