Search results for: immune organs
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 1174

Search results for: immune organs

334 Effect of Synthetic L-Lysine and DL-Methionine Amino Acids on Performance of Broiler Chickens

Authors: S. M. Ali, S. I. Mohamed

Abstract:

Reduction of feed cost for broiler production is at most importance in decreasing the cost of production. The objectives of this study were to evaluate the use of synthetic amino acids (L-lysine – DL-methionine) instead of super concentrate and groundnut cake versus meat powder as protein sources. A total of 180 male broiler chicks (Cobb – strain) at 15 day of age (DOA) were selected according to their average body weight (380 g) from a broiler chicks flock at Elbashair Farm. The chicks were randomly divided into six groups of 30 chicks. Each group was further sub divided into three replicates with 10 birds. Six experimental diets were formulated. The first diet contained groundnut cake and super concentrate as the control (GNC + C); in the second diet, meat powder and super concentrate (MP + C) were used. The third diet contained groundnut cake and amino acids (GNC + AA); the forth diet contained meat powder and amino acids (MP + AA). The fifth diet contained groundnut cake, meat powder and super concentrate (GNC + MP + C) and the sixth diet contained groundnut cake, meat powder and amino acids (GNC + MP + AA). The formulated rations were randomly assigned for the different sub groups in a completely randomized design of six treatments and three replicates. Weekly feed intake, body weight and mortality were recorded and body weight gain and feed conversion ratio were calculated. At the end of the experiment (49 DOA), nine birds from each treatment were slaughtered. Live body weight, carcass weight, head, shank, and some internal organs (gizzard, heart, liver, small intestine, and abdominal fat pad) weights were taken. For the overall experimental period the (GNC + C +MP) consumed significantly (P≤0.01) the highest cumulative feed while the (MP + AA) group consumed the lowest amount of feed. The (GNC + C) and the (GNC + AA) groups had the heaviest live body weight while (MP + AA) had the lowest live body weight. The overall FCR was significantly (P≤0.01) the best for (GNC + AA) group while the (MP + AA) reported the worst FCR. However, the (GNC + AA) had significantly (P≤0.01) the lowest AFP. The (GNC + MP + Con) group had the highest dressing % while the (MP + AA) group had the lowest dressing %. It is concluded that amino acids can be used instead of super concentrate in broiler feeding with perfect performance and less cost and that meat powder is not advisable to be used with amino acids.

Keywords: broiler chickens, DL-lysine, methionine, performance

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333 Glyco-Conjugated Gold Nanorods Based Biosensor for Optical Detection and Photothermal Ablation of Food Borne Bacteria

Authors: Shimayali Kaushal, Nitesh Priyadarshi, Nitin Kumar Singhal

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Food borne bacterial species have been identified as major pathogens in most of the severe pathogen-related diseases among humans which result in great loss to human health and food industry. Conventional methods like plating and enzyme-linked immune sorbent assay (ELISA) are time-consuming, laborious and require specialized instruments. Nanotechnology has emerged as a great field in case of rapid detection of pathogens in recent years. The AuNRs material has good electro-optical properties due to its larger light absorption band and scattering in surface plasmon resonance wavelength regions. By exploiting the sugar-based adhesion properties of microorganism, we can use the glycoconjugates capped gold nanorods as a potential nanobiosensor to detect the foodborne pathogen. In the present study, polyethylene glycol (PEG) coated gold nanorods (AuNRs) were prepared and functionalized with different types of carbohydrates and further characterized by UV-Visible spectrophotometry, dynamic light scattering (DLS), transmission electron microscopy (TEM). The reactivity of above said nano-biosensor was probed by lectin binding assay and also by different strains of foodborne bacteria by using spectrophotometric and microscopic techniques. Due to the specific interaction of probe with foodborne bacteria (Escherichia coli, Pseudomonas aeruginosa), our nanoprobe has shown significant and selective ablation of targeted bacteria. Our findings suggest that our nanoprobe can be an ideal candidate for selective optical detection of food pathogens and can reduce loss to the food industry.

Keywords: glyco-conjugates, gold nanorods, nanobiosensor, nanoprobe

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332 Effects of Hydroxysafflor Yellow a (HSYA) on UVA-Induced Damage in HaCaT Keratinocytes

Authors: Szu-Chieh Yu, Pei-Chin Chiand, Chih-Yi Lin, Yi-Wen Chien

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UV radiation from sunlight cause numbers of acute and chronic skin damage which can result in inflammation, immune changes, physical changes and DNA damage that facilitates skin aging and the development of skin carcinogenesis. Reactive oxygen species (ROS) are generated by excessive solar UV radiation, resulting in oxidative damage to cellar components, proteins, lipids, and nucleic acids. Thus, antioxidation plays an important role that protects skin against ROS-induced injury. Safflower (Carthamus tinctorius L.) is an important Chinese medicine contained abundance flavones and hydroxysafflor yellow A (HSYA) which is main active ingredient. HSYA is part of quinochalcone and has unique structures of hydroxy groups that provided the antioxidant effect. In this study, the aim was to investigate the protective role of HYSA in human keratinocytes (HaCaT) against UVA-induced oxidative damage and the possible mechanism. The HaCaT cells were UVA-irradiated and the effects of HYSA on cell viability, reactive oxygen species generation, DNA fragmentation and lipid peroxidation were measured. The mRNA expression of matrix metalloproteinase Ι (MMP Ι), cyclooxygenase-2 (COX-2) were determined by RT-PCR. In this study, UVA exposure lead to decrease in cell viability and increase in reactive oxygen species generation in HaCaT cells. HYSA could effectively increase the viability of HaCaT cells after UVA exposure and protect them from UVA-induced oxidative stress. Moreover, HYSA can reduce inflammation through inhibition the mRNA expression of MMP Ι and COX-2. Our results suggest that HSYA can act as a free radical scavenger while keratinocytes were photodamaged. HYSA could be a useful natural medicine for the protection of epidermal cells from UVA-induced damage and will be developed into products for skin care.

Keywords: HaCaT keratinocytes, hydroxysafflor yellow A (HSYA), MMP Ι, oxidative stress

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331 Impact of CYP3A5 Polymorphism on Tacrolimus to Predict the Optimal Initial Dose Requirements in South Indian Renal Transplant Recipients

Authors: S. Sreeja, Radhakrishnan R. Nair, Noble Gracious, Sreeja S. Nair, M. Radhakrishna Pillai

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Background: Tacrolimus is a potent immunosuppressant clinically used for the long term treatment of antirejection of transplanted organs in liver and kidney transplant recipients though dose optimization is poorly managed. However, So far no study has been carried out on the South Indian kidney transplant patients. The objective of this study is to evaluate the potential influence of a functional polymorphism in CYP3A5*3 gene on tacrolimus physiological availability/dose ratio in South Indian renal transplant patients. Materials and Methods: Twenty five renal transplant recipients receiving tacrolimus were enrolled in this study. Their body weight, drug dosage, and therapeutic concentration of Tacrolimus were observed. All patients were on standard immunosuppressive regime of Tacrolimus-Mycophenolate mofetil along with steroids on a starting dose of Tac 0.1 mg/kg/day. CYP3A5 genotyping was performed by PCR followed with RFLP. Conformation of RFLP analysis and variation in the nucleotide sequence of CYP3A5*3 gene were determined by direct sequencing using a validated automated generic analyzer. Results: A significant association was found between tacrolimus per dose/kg/d and CYP3A5 gene (A6986G) polymorphism in the study population. The CYP3A5 *1/*1, *1/*3 and *3/*3 genotypes were detected in 5 (20 %), 5 (20 %) and 15 (60 %) of the 25 graft recipients, respectively. CYP3A5*3 genotypes were found to be a good predictor of tacrolimus Concentration/Dose ratio in kidney transplant recipients. Significantly higher L/D was observed among non-expressors 9.483 ng/mL(4.5- 14.1) as compared with the expressors 5.154 ng/mL (4.42-6.5 ) of CYP3A5. Acute rejection episodes were significantly higher for CYP3A5*1 homozygotes compared to patients with CYP3A5*1/*3 and CYP3A5*3/*3 genotypes (40 % versus 20 % and 13 %, respectively ). The dose normalized TAC concentration (ng/ml/mg/kg) was significantly lower in patients having CYP3A5*1/*3 polymorphism. Conclusion: This is the first study to extensively determine the effect of CYP3A5*3 genetic polymorphism on tacrolimus pharmacokinetics in South Indian renal transplant recipients and also shows that majority of our patients carry mutant allele A6986G in CYP3A5*3 gene. Identification of CYP3A5 polymorphism prior to transplantation could contribute to evaluate the appropriate initial dosage of tacrolimus for each patient.

Keywords: kidney transplant patients, CYP3A5 genotype, tacrolimus, RFLP

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330 Aberrant Genome‐Wide DNA Methylation Profiles of Peripheral Blood Mononuclear Cells from Patients Hospitalized with COVID-19

Authors: Inam Ridha, Christine L. Kuryla, Madhuranga Thilakasiri Madugoda Ralalage Don, Norman J. Kleiman, Yunro Chung, Jin Park, Vel Murugan, Joshua LaBaer

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To date, more than 275 million people worldwide have been diagnosed with COVID-19 and the rapid spread of the omicron variant suggests many millions more will soon become infected. Many infections are asymptomatic, while others result in mild to moderate illness. Unfortunately, some infected individuals exhibit more serious symptoms including respiratory distress, thrombosis, cardiovascular disease, multi-organ failure, cognitive difficulties, and, in roughly 2% of cases, death. Studies indicate other coronaviruses can alter the host cell's epigenetic profile and lead to alterations in the immune response. To better understand the mechanism(s) by which SARS-CoV-2 infection causes serious illness, DNA methylation profiles in peripheral blood mononuclear cells (PBMCs) from 90 hospitalized severely ill COVID-19 patients were compared to profiles from uninfected control subjects. Exploratory epigenome-wide DNA methylation analyses were performed using multiplexed methylated DNA immunoprecipitation (MeDIP) followed by pathway enrichment analysis. The findings demonstrated significant DNA methylation changes in infected individuals as compared to uninfected controls. Pathway analysis indicated that apoptosis, cell cycle control, Toll-like receptors (TLR), cytokine interactions, and T cell differentiation were among the most affected metabolic processes. In addition, changes in specific gene methylation were compared to SARS-CoV-2 induced changes in RNA expression using published RNA-seq data from 3 patients with severe COVID-19. These findings demonstrate significant correlations between differentially methylated and differentially expressed genes in a number of critical pathways.

Keywords: COVID19, epigenetics, DNA mathylation, viral infection

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329 Study on Accumulation of Heavy Metals in Sweet Potato, Grown in Industrially Polluted Regions

Authors: Violina Angelova, Galina Pevicharova

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A comparative research had been carried out to allow us to determine the quantities and the centers of accumulation of Pb, Cu, Zn and Cd in the vegetative and reproductive organs of the sweet potatoes and to ascertain the possibilities for growing them on soils, polluted with heavy metals. The experiments were performed on agricultural fields contaminated by the (1) Non-Ferrous-Metal Works near Plovdiv, (2) Lead and Zinc Complex near Kardjali and (3) a copper smelter near Pirdop, Bulgaria. The soils used in this experiment were characterized by acid, neutral and slightly alkaline reaction, loamy texture and a moderate content of organic matter. The total content of Zn, Pb, and Cd was high and exceeded the limit value in agriculture soils. Sweet potatoes were in a 2-year rotation scheme on three blocks in the experimental field. On reaching commercial ripeness the sweet potatoes were gathered and the contents of heavy metals in their different parts – root, tuber (peel and core), leaves and stems, were determined after microwave mineralization. The quantitative measurements were carried out with inductively coupled plasma atomic emission spectroscopy. The contamination of the sweet potatoes was due mainly to the presence of heavy metals in the soil, which entered the plants through their root system, as well as by diffusion through the peel. Pb, Cu, Zn, and Cd were selectively accumulated in the underground parts of the sweet potatoes, and most of all in the root system and the peel. Heavy metals have an impact on the development and productivity of the sweet potatoes. The high anthropogenic contamination leads to an increased assimilation of heavy metals which reduces the yield and the quality of the production of sweet potatoes, as well as leads to decrease of the absolute dry substance and the quantity of sugars in sweet potatoes. Sweet potatoes could be grown on soils, which are light to medium polluted with lead, zinc, and cadmium, as they do not accumulate these elements. On heavily polluted soils, however, (Pb – 1504 mg/kg, Zn – 3322 mg/kg, Cd – 47 mg/kg) the growing of sweet potatoes is not allowed, as the accumulation of Pb and Cd in the core of the potatoes exceeds the Maximum Acceptable Concentration. Acknowledgment: The authors gratefully acknowledge the financial support by the Bulgarian National Science Fund (Project DFNI DH04/9).

Keywords: heavy metals, polluted soils, sweet potatoes, uptake

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328 Effect of Supplementing Different Sources and Levels of Phytase Enzyme to Diets on Productive Performance for Broiler Chickens

Authors: Sunbul Jassim Hamodi, Muna Khalid Khudayer, Firas Muzahem Hussein

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The experiment was conducted to study the effect of supplement sources of Phytase enzyme (bacterial, fungal, enzymes mixture) using levels (250, 500, 750) FTY/ kg feed to diets compared with control on the performance for one thousand fifty broiler chicks (Ross 308) from 1day old with initial weight 39.78 gm till 42 days. The study involved 10 treatments, three replicates per treatment (35 chicks/replicate). Treatments were as follows: T1: control diet (without any addition). T2: added bacterial phytase enzyme 250FTY/ kg feed. T3: added bacterial phytase enzyme 500FTY/ kg feed. T4: added bacterial phytase enzyme 750FTY/ kg feed. T5: added fungal phytase enzyme 250FTY/ kg feed. T6: added fungal phytase enzyme 500FTY/ kg feed. T7: added fungal phytase enzyme 750FTY/ kg feed. T8 added enzymes mixture 250U/ kg feed. T9: added enzymes mixture 500U/ kg feed. T10: added enzymes mixture 750U/ kg feed. The results revealed that supplementing 750 U from enzymes mixture to broiler diet increased significantly (p <0.05) body weight compared with (250 FTY bacterial phytase/Kgfeed), (750 FTY bacterial phytase/Kg feed), (750FTY fungal phytase/Kgfeed) at 6 weeks, also supplemented different sources and levels from phytase enzyme improved a cumulative weight gain for (500 FTY bacterial phytase/Kgfeed), (250FTY fungal phytase/Kgfeed), (500FTY fungal phytase/Kgfeed), (250 Uenzymes mixture/Kgfeed), (500 Uenzymes mixture/Kgfeed) and (750 U enzymes mixture/Kgfeed) treatments compared with (750 FTY fungal phytase/Kgfeed)treatment, about accumulative feed consumption (500 FTY fungal phytase/Kgfeed) and (250 Uenzymes mixture/Kgfeed) increased significantly compared with control group and (750FTY fungal phytase/Kgfeed) during 1-6 weeks. There were significantly improved in cumulative feed conversion for (500U enzymes mixture/Kgfeed) compared with the worse feed conversion ratio that recorded in (250 FTY bacterial phytase/Kgfeed). No significant differences between treatments in internal organs relative weights, carcass cuts, dressing percentage and production index. Mortality was increased in (750FTY fungal phytase/Kgfeed) compared with other treatments.

Keywords: phytase, phytic acid, broiler, productive performance

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327 Redirection of Cytokine Production Patterns by Dydrogesterone, an Orally-Administered Progestogen

Authors: Raj Raghupathy

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Recurrent Spontaneous Miscarriage (RSM) is a common form of pregnancy loss, 50% of which are due to ‘unexplained’ causes. Evidence exists to suggest that RSM may be caused by immunologic factors such as cytokines which are critical molecules of the immune system, with an impressive array of capabilities. An association appears to exist between Th2-type reactivity (mediated by Th2 or anti-inflammatory cytokines) and normal, successful pregnancy, and between unexplained RSM and Th1 cytokine dominance. If pro-inflammatory cytokines are indeed associated with pregnancy loss, the suppression of these cytokines, and thus the ‘redirection’ of maternal reactivity, may help prevent cytokine-mediated pregnancy loss. The objective of this study was to explore the possibility of modulating cytokine production using Dydrogesterone (Duphaston®), an orally-administered progestogen. Peripheral blood mononuclear cells from 34 women with a history of at least 3 unexplained recurrent miscarriages were stimulated in vitro with a mitogen (to elicit cytokine production) in the presence and absence of dydrogesterone. Levels of selected pro- and anti-inflammatory cytokines produced by peripheral blood mononuclear cells were measured after exposure to these progestogens. Dydrogesterone down-regulates the production of pro-inflammatory cytokines and up-regulates the production of anti-inflammatory cytokines. The ratios of Th2 to Th1 cytokines are markedly elevated in the presence of dydrogesterone, indicating a shift from potentially harmful maternal Th1 reactivity to a more pregnancy-conducive Th2 profile. We used a progesterone receptor antagonist to show that this cytokine-modulating effect of dydrogesterone is mediated via the progesterone receptor. Dydrogesterone also induces the production of the Progesterone-Induced Blocking Factor (PIBF); lymphocytes exposed to PIBF produce higher levels of Th2 cytokines, affecting a Th1 → Th2 cytokine shift which could be favourable to the success of pregnancy. We conclude that modulation of maternal cytokine production profiles is possible with dydrogesterone which has the merits that it can be administered orally and that it is safe.

Keywords: cytokines, dydrogesterone, progesterone, recurrent spontaneous miscarriage

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326 Human Endogenous Retrovirus Link With Multiple Sclerosis Disease Progression

Authors: Sina Mahdavi

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Background and Objective: Multiple sclerosis (MS) is an inflammatory autoimmune disease of the CNS that affects the myelination process in the central nervous system (CNS). Complex interactions of various "environmental or infectious" factors may act as triggers in autoimmunity and disease progression. The association between viral infections, especially human endogenous retrovirus (HERV) and MS is one potential cause that is not well understood. This study aims to summarize the available data on HERV infection in MS disease progression. Materials and Methods: For this study, the keywords "Multiple sclerosis", "Human endogenous retrovirus", and "central nervous system" in the databases PubMed, Google Scholar, Sid, and MagIran between 2016 and 2022 were searched and 14 articles chosen, studied, and analyzed. Results: In the leptomeningeal cells of MS patients, a retrovirus-like element associated with reverse transcriptase (RT) activity called multiple sclerosis-associated retroviruses (MSRV) has been identified. HERVs are expressed in the human CNS despite mechanisms to suppress their expression. External factors, especially viral infections such as influenza virus, Epstein-Barr virus, and herpes simplex virus type 1, can activate HERV gene expression. The MSRV coat protein is activated by activating TLR4 at the brain surface, particularly in oligodendroglial progenitor cells and macrophages, leading to immune cascades followed by the downregulation of myelin protein expression. The HERV-K18 envelope gene (env) acts as a superantigen and induces inflammatory responses in patients with MS. Conclusion: There is a high expression of endogenous retroviruses during the course of MS, which indicates the relationship between HERV and MS, that this virus can play a role in the development of MS by creating an inflammatory state. Therefore, measures to modulate the expression of endogenous retroviruses may be effective in reducing inflammatory processes in demyelinated areas of MS patients.

Keywords: multiple sclerosis, human endogenous retrovirus, central nervous system, MSRV

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325 Molecular Pathogenesis of NASH through the Dysregulation of Metabolic Organ Network in the NASH-HCC Model Mouse Treated with Streptozotocin-High Fat Diet

Authors: Bui Phuong Linh, Yuki Sakakibara, Ryuto Tanaka, Elizabeth H. Pigney, Taishi Hashiguchi

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NASH is an increasingly prevalent chronic liver disease that can progress to hepatocellular carcinoma and now is attracting interest worldwide. The STAM™ model is a clinically-correlated murine NASH model which shows the same pathological progression as NASH patients and has been widely used for pharmacological and basic research. The multiple parallel hits hypothesis suggests abnormalities in adipocytokines, intestinal microflora, and endotoxins are intertwined and could contribute to the development of NASH. In fact, NASH patients often exhibit gut dysbiosis and dysfunction in adipose tissue and metabolism. However, the analysis of the STAM™ model has only focused on the liver. To clarify whether the STAM™ model can also mimic multiple pathways of NASH progression, we analyzed the organ crosstalk interactions between the liver and the gut and the phenotype of adipose tissue in the STAM™ model. NASH was induced in male mice by a single subcutaneous injection of 200 µg streptozotocin 2 days after birth and feeding with high-fat diet after 4 weeks of age. The mice were sacrificed at NASH stage. Colon samples were snap-frozen in liquid nitrogen and stored at -80˚C for tight junction-related protein analysis. Adipose tissue was prepared into paraffin blocks for HE staining. Blood adiponectin was analyzed to confirm changes in the adipocytokine profile. Tight junction-related proteins in the intestine showed that expression of ZO-1 decreased with the progression of the disease. Increased expression of endotoxin in the blood and decreased expression of Adiponectin were also observed. HE staining revealed hypertrophy of adipocytes. Decreased expression of ZO-1 in the intestine of STAM™ mice suggests the occurrence of leaky gut, and abnormalities in adipocytokine secretion were also observed. Together with the liver, phenotypes in these organs are highly similar to human NASH patients and might be involved in the pathogenesis of NASH.

Keywords: Non-alcoholic steatohepatitis, hepatocellular carcinoma, fibrosis, organ crosstalk, leaky gut

Procedia PDF Downloads 157
324 Newspaper Coverage and the Prevention of Child Sexual Abuse in Nigeria

Authors: Grace Iember Anweh, Er Shipp

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Child Sexual Abuse (CSA) has been a contending issue across the globe. The menace of child sexual violence cuts across all continents. From 0 - 13 years, children have been sexually abused – some to the extent that their reproductive organs have been permanently damaged. The research in view is timely, as it will contribute data on CSA and media role to the communication parlance. This study believes that the adverse effects of this menace can hinder children who are potential leaders of tomorrow from harnessing their potentials to contribute to the growth and development of societies due to the psychological, health, and social effects of sex abuse. Where government policies, the law, cultural beliefs, and bottle necks surrounding processes of fighting child sexual abuse have failed, this study assumes that adequate coverage by the mass media, especially the newspapers known for their in-depth coverage and reporting, can help to eradicate or reduce to its barest minimum, the menace of CSA. Therefore, this study aims at assessing the coverage of newspapers – their policies and content towards preventive strategies, and how the public access and receive the messages to the extent they take action to forestall the persistence of sexual violation of children in Nigeria. Methodologically, the study has adopted qualitative and quantitative methods to answer the problem. The study used in-depth interview method to find out from journalists and editors of newspapers the policies that define the production of news content on sexual gender-based violence. In addition, selected National Daily newspapers are content - analysed to determine the focus of media coverage and whether the contents are preventive-based or case-based. In addition, caregivers of the reproductive ages from 16 years and above, ranging from parents, guardians, and school management, will form the study population through a survey using the questionnaire. The aim is to determine their views regarding mass media coverage of sexual violence against children and the effectiveness of the content, to the extent of prompting them to keep the child safe from sexual molesters. Findings from the content analysis so far show that newspapers in Nigeria are not engaged in preventive content of CSA. Their contents are rather case-based.

Keywords: newspaper, coverage, prevention, child, sexual abuse

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323 Performance of Non-Deterministic Structural Optimization Algorithms Applied to a Steel Truss Structure

Authors: Ersilio Tushaj

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The efficient solution that satisfies the optimal condition is an important issue in the structural engineering design problem. The new codes of structural design consist in design methodology that looks after the exploitation of the total resources of the construction material. In recent years some non-deterministic or meta-heuristic structural optimization algorithms have been developed widely in the research community. These methods search the optimum condition starting from the simulation of a natural phenomenon, such as survival of the fittest, the immune system, swarm intelligence or the cooling process of molten metal through annealing. Among these techniques the most known are: the genetic algorithms, simulated annealing, evolution strategies, particle swarm optimization, tabu search, ant colony optimization, harmony search and big bang crunch optimization. In this study, five of these algorithms are applied for the optimum weight design of a steel truss structure with variable geometry but fixed topology. The design process selects optimum distances and size sections from a set of commercial steel profiles. In the formulation of the design problem are considered deflection limitations, buckling and allowable stress constraints. The approach is repeated starting from different initial populations. The design problem topology is taken from an existing steel structure. The optimization process helps the engineer to achieve good final solutions, avoiding the repetitive evaluation of alternative designs in a time consuming process. The algorithms used for the application, the results of the optimal solutions, the number of iterations and the minimal weight designs, will be reported in the paper. Based on these results, it would be estimated, the amount of the steel that could be saved by applying structural analysis combined with non-deterministic optimization methods.

Keywords: structural optimization, non-deterministic methods, truss structures, steel truss

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322 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

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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

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321 Ecofriendly Approach for the Management of Red Cotton Bug Dysdercus koenigii by Botanicals

Authors: S: Kayesth, K. K. Gupta

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The indiscriminate use of insecticides causes environmental contamination, adversely affects non-target organisms and develops resistance among insects and pests. There has always been felt a need for methods of control which can overcome these environmental and other ecological issues. The present study was designed to evaluate the effect of different plants volatiles on survival, longevity, growth, development and reproduction of Dysdercus koenigii. The hexane extract of three different plants (Catharanthus roseus, Ocimum sanctum and Lantana camara) was used. The fifth instars were exposed to hexane extract with concentrations of 10%, 5%, 2.5%, 1.25%, 0.1%, 0.5%, 0.25%, 0.13% and 0.06% while adults were treated with 10%, 5%, 2.5% and 1.25%. 1-ml of each of these concentrations was used to make a thin film in sterilized glass jars of 500 ml capacity. Fifteen newly emerged fifth instar nymphs and ten pairs of adult bugs were treated separately with the extracts for 24 hour exposure to the plant volatiles. The effect of these plant extract was observed and readings were recorded for 23 days. Survival and longevity of both fifth instars and adults were in correlation with the concentrations of the plant extracts. The extracts did not influence growth of fifth instars significantly but impaired their development significantly at higher concentrations. The treated nymphs at higher concentrations either could not moult or died and those which could moult moulted into supranumery instars, adultoids or adults with wing deformities. The supranumery insects retained the nymphal characters except increased body size and wing pads. The adultoids had wing deformities and non-functional reproductive organs. Adultoids exhibited courtship and mounting attempts but were not able to mate. At lower concentrations from 0.1 to 0.06% the fifth instars developed into adults with fewer deformities. At these concentrations, the fecundity and fertility of these adults were drastically reduced. On the contrary, the treated adults also had reduced fecundity and fertility compared to control. Among three plant extracts Ocimcum was most toxic for both fifth instars and adults in terms of survival and longevity. Catharanthus, Ocimum and Lantana appeared to have potential molecules which possessed insect juvenile hormone like activity. Potential application of these plant extracts in IPM was discussed.

Keywords: Catharanthus, Ocimum, Lantana, Dysdercus koenigii

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320 Companies and Transplant Tourists to China

Authors: Pavel Porubiak, Lukas Kudlacek

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Introduction Transplant tourism is a controversial method of obtaining an organ, and that goes all the more for a country such as China, where sources of evidence point out to the possibility of organs being harvested illegally. This research aimed at listing the individual countries these tourists come from, or which medical companies sell transplant related products in there, with China being used as an example. Materials and methods The methodology of scoping study was used for both parts of the research. The countries from which transplant tourists come to China were identified by a search through existing medical studies in the NCBI PubMed database, listed under the keyword ‘transplantation in China’. The search was not limited by any other criteria, but only the studies available for free – directly on PubMed or a linked source – were used. Other research studies on this topic were considered as well. The companies were identified through multiple methods. The first was an online search focused on medical companies and their products. The Bloomberg Service, used by stock brokers worldwide, was then used to identify the revenue of these companies in individual countries – if data were available – as well as their business presence in China. A search through the U.S. Securities and Exchange Commission was done in the same way. Also a search on the Chinese internet was done, and to obtain more results, a second online search was done as well. The results and discussion The extensive search has identified 14 countries with transplant tourists to China. The search for a similar studies or reports resulted in finding additional six countries. The companies identified by our research also amounted to 20. Eight of them are sourcing China with organ preservation products – of which one is just trying to enter the Chinese market, six with immunosuppressive drugs, four with transplant diagnostics, one with medical robots which Chinese doctors use for transplantation as well, and another one trying to enter the Chinese market with a consumable-type product also related to transplantation. The conclusion The question of the ethicality of transplant tourism may be very pressing, since as the research shows, just the sheer amount of participating countries, sourcing transplant tourists to another one, amounts to 20. The identified companies are facing risks due to the nature of transplantation business in China, as officially executed prisoners are used as sources, and widely cited pieces of evidence point out to illegal organ harvesting. Similar risks and ethical questions are also relevant to the countries sourcing the transplant tourists to China.

Keywords: China, illegal organ harvesting, transplant tourism, organ harvesting technology

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319 Immunomodulatory Activity of Polysaccharide-Protein Complex Isolated from the Sclerotia of Polyporus Rhinocerus in Murine Macrophages

Authors: Chaoran Liu

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Bioactive polysaccharides and polysaccharide-protein complex derived from mushrooms and fungi have a wide range of immunomodulatory activity with low side-effects and have therefore the potential to be developed as an adjuvant in cancer therapies. Mushrooms sclerotium is rich in polysaccharides and the polysaccharides isolated from the sclerotium of Polyporus rhinocerus have shown potent in vivo and in vitro immunomodulatory effects. Macrophages are considered to be an important component of the innate immune response against bacterial infection and cancer. To better understanding the immunomodulatory effects and its underlying mechanisms of sclerotial water-soluble polysaccharides extracted from P. rhinocerus on macrophages, the objectives of this study are to purify the water-soluble novel sclerotial polysaccharides and to characterize the structure and properties as well as to study the detailed molecular mechanisms of the in vitro immunomodulating effects in murine macrophages. The hot water-soluble fraction PRW from the sclerotium of P. rhinocerus was obtained using solvent extraction. PRW was further fractionated by membrane ultrafiltration to a give a fraction (PRW1) with molecular mass less than 50 kDa. PRW1 was characterized to be a polysaccharide-protein complex composed of 45.7% polysaccharide and 44.2% protein. The chemical structure of the carbohydrate moiety of PRW1 was elucidated by GC and FTIR to be mainly beta-D-glucan with trace amount of galactose and mannose. The immunomodulatory effects of PRW1 on murine RAW 264.7 macrophages were demonstrated in terms of the increase in nitric oxide production and cytokine production. Mechanistically, PRW1 initiates ERK phosphorylation to activate macrophages within 15 min and significantly improves the expression level of inducible NOS (iNOS) from 6 h after treatment. In summary, this study indicates that PRW1 is a potent immunomodulatory agent for macrophages and suggests that mushroom sclerotia from Polyporus rhinocerus requires for further investigation in cancer research.

Keywords: Polyporus rhinocerus, mushroom sclerotia, Polysaccharide-Protein Complex, macrophage activation

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318 Effect of Co-Infection With Intestinal Parasites on COVID-19 Severity: A Prospective Observational Cohort Study

Authors: Teklay Gebrecherkos, Dawit Wolday, Muhamud Abdulkader

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Background: COVID-19 symptomatology in Africa appears significantly less serious than in the industrialized world. Our hypothesis for this phenomenon, being a different, more activated immune system due to parasite infections contributes to reduced COVID-19 outcome. We investigated this hypothesis in an endemic area in sub sub-saharan Africa. Methods: Ethiopian COVID-19 patients were enrolled and screened for intestinal parasites, between July 2020 and March 2021. The primary outcome was the proportion of patients with severe COVID-19. SARS-CoV-2 infection was confirmed by RT-PCR on samples obtained from nasopharyngeal swabs, while direct microscopic examination, modified Ritchie concentration, and Kato-Katz methods were used to identify parasites and ova from a fresh stool sample. Ordinal logistic regression models were used to estimate the association between parasite infection and COVID-19 severity. Models were adjusted for sex, age, residence, education level, occupation, body mass index, and comorbidities. Data were analyzed using STATA version 14. P-value <0.05 was considered statistically significant. Results: A total of 751 SARS-CoV-2 infected patients were enrolled, of whom 284 (37•8%) had an intestinal parasitic infection. Only 27/255 (10•6%) severe COVID-19 patients were co-infected with intestinal parasites, while 257/496 (51•8%) non-severe COVID-19 patients appeared parasite positive (p<0.0001). Patients co-infected with parasites had lower odds of developing severe COVID-19, with an adjusted odds ratio (AOR) of 0•14 (95% CI 0•09–0•24; p<0•0001) for all parasites, AOR 0•20 ([95% CI 0•11–0•38]; p<0•0001) for protozoa, and AOR 0•13 ([95% CI 0•07–0•26]; p<0•0001) for helminths. When stratified by species, co-infection with Entamoeba spp., Hymenolopis nana, and Schistosoma mansoni implied a lower probability of developing severe COVID-19. There were 11 deaths (1•5%), and all were among patients without parasites (p=0•009). Conclusions: Parasite co-infection is associated with a reduced risk of severe COVID-19 in African patients. Parasite-driven immunomodulatory responses may mute hyper-inflammation associated with severe COVID-19.

Keywords: COVID-19, SARS-COV-2, intestinal parasite, RT-PCR, co-infection

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317 Carbendazim Toxicity and Ameliorative Effect of Vitamin E in African Giant Rats

Authors: A. O. Omonona, T. A. Jarikre

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Increase specialization in agriculture and use of pesticides may inadvertently cause ecosystem degradation and eventually loss of biodiversity. The populations of numerous wildlife species have undergone a precipitous decline. Many of these problems have been attributed directly to habitat loss and over exploitation resulting from unregulated pesticide uses. Carbendazim a broad spectrum benzimidazole fungicide and a metabolite of benomyl, is used to control plant disease in cereals and fruit. The effect of carbendazim exposure and the ameliorative effect of tocopherol (vitamin E) were assessed on African giant rat AGR. Hematological, biochemical and histological changes were used to determine the health condition of the animals exposed to pesticide. Sixteen AGR were stabilized, weighed and then divided into four experimental groups (A to D). Two groups were pretreated with vitamin. Group A was exposed to carbendazim only, B- carbendazim + vitamin, C- vitamin only, and D- blank (control). Packed cell volume PCV was estimated by the microhematocrit method, Leucocyte and Platelet counts were determined using the hemocytometric method. Cholinesterase (AchE) and markers of oxidative stress were quantified, and tissue changes examined microscopically. There were no behavioral changes observed in the animals, but there was a decrease in body weight and abortion after 23 days of exposure to carbendazim. There was significant differences in the packed cell volume, the hemoglobin concentration and the red blood cell counts (p < 0.05). The increases in malonyl aldehyde MDA was significant (p < 0.05) in the pesticide intoxicated rats compared to control. Vitamin E supplementation reduced MDA level significantly (p < 0.05). There was a sharp remarkable decrease in acetylcholinesterase levels in the pesticide intoxicated rats (p < 0.05). Vitamin E supplementation normalise the AchE levels comparable to that in control. Grossly, the vital organs appeared normal in the pesticide exposed and control groups except moderate pulmonary congestion. Microscopically, there was severe diffuse hepatocellular swelling in carbendazim exposed group. The severity of hepatocellular injury was reduced in the rats with vitamin E. This study ascertained the toxic effect of carbendazim and antioxidative properties of vitamins in the Africa giant rat.

Keywords: African giant rat, antioxidant, carbendazim, pesticides, toxicity

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316 Transcriptional Evidence for the Involvement of MyD88 in Flagellin Recognition: Genomic Identification of Rock Bream MyD88 and Comparative Analysis

Authors: N. Umasuthan, S. D. N. K. Bathige, W. S. Thulasitha, I. Whang, J. Lee

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The MyD88 is an evolutionarily conserved host-expressed adaptor protein that is essential for proper TLR/ IL1R immune-response signaling. A previously identified complete cDNA (1626 bp) of OfMyD88 comprised an ORF of 867 bp encoding a protein of 288 amino acids (32.9 kDa). The gDNA (3761 bp) of OfMyD88 revealed a quinquepartite genome organization composed of 5 exons (with the sizes of 310, 132, 178, 92 and 155 bp) separated by 4 introns. All the introns displayed splice signals consistent with the consensus GT/AG rule. A bipartite domain structure with two domains namely death domain (24-103) coded by 1st exon, and TIR domain (151-288) coded by last 3 exons were identified through in silico analysis. Moreover, homology modeling of these two domains revealed a similar quaternary folding nature between human and rock bream homologs. A comprehensive comparison of vertebrate MyD88 genes showed that they possess a 5-exonic structure. In this structure, the last three exons were strongly conserved, and this suggests that a rigid structure has been maintained during vertebrate evolution. A cluster of TATA box-like sequences were found 0.25 kb upstream of cDNA starting position. In addition, putative 5'-flanking region of OfMyD88 was predicted to have TFBS implicated with TLR signaling, including copies of NFB1, APRF/ STAT3, Sp1, IRF1 and 2 and Stat1/2. Using qPCR technique, a ubiquitous mRNA expression was detected in liver and blood. Furthermore, a significantly up-regulated transcriptional expression of OfMyD88 was detected in head kidney (12-24 h; >2-fold), spleen (6 h; 1.5-fold), liver (3 h; 1.9-fold) and intestine (24 h; ~2-fold) post-Fla challenge. These data suggest a crucial role for MyD88 in antibacterial immunity of teleosts.

Keywords: MyD88, innate immunity, flagellin, genomic analysis

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315 Control of Lymphatic Remodelling by miR-132

Authors: Valeria Arcucci, Musarat Ishaq, Steven A. Stacker, Greg J. Goodall, Marc G. Achen

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Metastasis is the lethal aspect of cancer for most patients. Remodelling of lymphatic vessels associated with a tumour is a key initial step in metastasis because it facilitates the entry of cancer cells into the lymphatic vasculature and their spread to lymph nodes and distant organs. Although it is clear that vascular endothelial growth factors (VEGFs), such as VEGF-C and VEGF-D, are key drivers of lymphatic remodelling, the means by which many signaling pathways in endothelial cells are coordinately regulated to drive growth and remodelling of lymphatics in cancer is not understood. We seek to understand the broader molecular mechanisms that control cancer metastasis, and are focusing on microRNAs, which coordinately regulate signaling pathways involved in complex biological responses in health and disease. Here, using small RNA sequencing, we found that a specific microRNA, miR-132, is upregulated in expression in lymphatic endothelial cells (LECs) in response to the lymphangiogenic growth factors. Interestingly, ectopic expression of miR-132 in LECs in vitro stimulated proliferation and tube formation of these cells. Moreover, miR-132 is expressed in lymphatic vessels of a subset of human breast tumours which were previously found to express high levels of VEGF-D by immunohistochemical analysis on tumour tissue microarrays. In order to dissect the complexity of regulation by miR-132 in lymphatic biology, we performed Argonaute HITS-CLIP, which led us to identify the miR-132-mRNA interactome in LECs. We found that this microRNA in LECs is involved in the control of many different pathways mainly involved in cell proliferation and regulation of the extracellular matrix and cell-cell junctions. We are now exploring the functional significance of miR-132 targets in the biology of LECs using biochemical techniques, functional in vitro cell assays and in vivo lymphangiogenesis assays. This project will ultimately define the molecular regulation of lymphatic remodelling by miR-132, and thereby identify potential therapeutic targets for drugs designed to restrict the growth and remodelling of tumour lymphatics resulting in metastatic spread.

Keywords: argonaute HITS-CLIP, cancer, lymphatic remodelling, miR-132, VEGF

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314 Treating Complex Pain and Addictions with Bioelectrode Therapy: An Acupuncture Point Stimulus Method for Relieving Human Suffering

Authors: Les Moncrieff

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In a world awash with potent opioids flaming an international crisis, the need to explore safe alternatives has never been more urgent. Bio-electrode Therapy is a novel adjunctive treatment method for relieving acute opioid withdrawal symptoms and many types of complex acute and chronic pain (often the underlying cause of opioid dependence). By combining the science of developmental bioelectricity with Traditional Chinese Medicine’s theory of meridians, rapid relief from pain is routinely being achieved in the clinical setting. Human body functions are dependent on electrical factors, and acupuncture points on the body are known to have higher electrical conductivity than surrounding skin tissue. When tiny gold- and silver-plated electrodes are secured to the skin at specific acupuncture points using established Chinese Medicine principles and protocols, an enhanced microcurrent and electrical field are created between the electrodes, influencing the entire meridian and connecting meridians. No external power source or electrical devices are required. Endogenous DC electric fields are an essential fundamental component for development, regeneration, and wound healing. Disruptions in the normal ion-charge in the meridians and circulation of blood will manifest as pain and development of disease. With the application of these simple electrodes (gold acting as cathode and silver as anode) according to protocols, the resulting microcurrent is directed along the selected meridians to target injured or diseased organs and tissues. When injured or diseased cells have been stimulated by the microcurrent and electrical fields, the permeability of the cell membrane is affected, resulting in an immediate relief of pain, a rapid balancing of positive and negative ions (sodium, potassium, etc.) in the cells, the restoration of intracellular fluid levels, replenishment of electrolyte levels, pH balance, removal of toxins, and a re-establishment of homeostasis.

Keywords: bioelectricity, electrodes, electrical fields, acupuncture meridians, complex pain, opioid withdrawal management

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313 Hydrogel Hybridizing Temperature-Cured Dissolvable Gelatin Microspheres as Non-Anchorage Dependent Cell Carriers for Tissue Engineering Applications

Authors: Dong-An Wang

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All kinds of microspheres have been extensively employed as carriers for drug, gene and therapeutic cell delivery. Most therapeutic cell delivery microspheres rely on a two-step methodology: fabrication of microspheres and subsequent seeding of cells onto them. In this study, we have developed a novel one-step cell encapsulation technique using a convenient and instant water-in-oil single emulsion approach to form cell-encapsulated gelatin microspheres. This technology is adopted for hyaline cartilage tissue engineering, in which autologous chondrocytes are used as therapeutic cells. Cell viability was maintained throughout and after the microsphere formation (75-100 µm diameters) process that avoids involvement of any covalent bonding reactions or exposure to any further chemicals. Further encapsulation of cell-laden microspheres in alginate gels were performed under 4°C via a prompt process. Upon the formation of alginate constructs, they were immediately relocated into CO2 incubator where the temperature was maintained at 37°C; under this temperature, the cell-laden gelatin microspheres dissolved within hours to yield similarly sized cavities and the chondrocytes were therefore suspended within the cavities inside the alginate gel bulk. Hence, the gelatin cell-laden microspheres served two roles: as cell delivery vehicles which can be removable through temperature curing, and as porogens within an alginate hydrogel construct to provide living space for cell growth and tissue development as well as better permeability for mutual diffusions. These cell-laden microspheres, namely “temperature-cured dissolvable gelatin microsphere based cell carriers” (tDGMCs), were further encapsulated in a chondrocyte-laden alginate scaffold system and analyzed by WST-1, gene expression analyses, biochemical assays, histology and immunochemistry stains. The positive results consistently demonstrated the promise of tDGMC technology in delivering these non-anchorage dependent cells (chondrocytes). It can be further conveniently translated into delivery of other non-anchorage dependent cell species, including stem cells, progenitors or iPS cells, for regeneration of tissues in internal organs, such as engineered hepatogenesis or pancreatic regeneration.

Keywords: biomaterials, tissue engineering, microsphere, hydrogel, porogen, anchorage dependence

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312 Pellet Feed Improvements through Vitamin C Supplementation for Snakehead (Channa striata) Culture in Vietnam

Authors: Pham Minh Duc, Tran Thi Thanh Hien, David A. Bengtson

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Laboratory feeding trial: the study was conducted to find out the optimal dietary vitamin C, or ascorbic acid (AA) levels in terms of the growth performance of snakehead. The growth trial included six treatments with five replications. Each treatment contained 0, 125, 250, 500, 1000 and 2000 mg AA equivalent kg⁻¹ diet which included six iso-nitrogenous (45% protein), iso-lipid (9% lipid) and isocaloric (4.2 Kcal.g¹). Eighty snakehead fingerlings (6.24 ± 0.17 g.fish¹) were assigned randomly in 0.5 m³ composite tanks. Fish were fed twice daily on demand for 8 weeks. The result showed that growth rates increased, protein efficiency ratio increased and the feed conversion ratio decreased in treatments with AA supplementation compared with control treatment. The survival rate of fish tends to increase with increase AA level. The number of RBCs, lysozyme in treatments with AA supplementation tended to rise significantly proportional to the concentration of AA. The number of WBCs of snakehead in treatments with AA supplementation was higher 2.1-3.6 times. In general, supplementation of AA in the diets for snakehead improved growth rate, feed efficiency and immune response. Hapa on-farm trial: based on the results of the laboratory feeding trial, the effects of AA on snakehead in hapas to simulate farm conditions, was tested using the following treatments: commercial feed; commercial feed plus hand mixed AA at 500; 750 and 1000 mg AA.kg⁻¹; SBM diet without AA; SBM diet plus 500; 750 and 1000 mg AA.kg⁻¹. The experiment was conducted in two experimental ponds (only SBM diet without AA placed in one pond and the rest in the other pond) with four replicate hapa each. Stocking density was 150 fish.m² and culture period was 5 months until market size was attained. The growth performance of snakehead and economic aspects were examined in this research.

Keywords: fish health, growth rate, snakehead, Vitamin C

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311 Prevalence and Distribution of Verocytotoxigenic Escherichia coli (Vtec) Non-O157 Serotypes in Cattle in Abuja, Nigeria

Authors: S. I. Enem, S. I. Oboegbulem

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Objective: The most frequently implicated E. coli serotype causing haemorrhagic colitis and haemorrhagic uraemic syndrome (HUS) is VTEC 0157. However, non-O157 VTEC is now known to be as prevalent as VETC O157 infection (or even more) in most parts of the world. The objective of the study was to establish the occurrence of non-O157 VTEC serotypes in cattle in the Federal Capital Territory (FCT) Abuja, Nigeria. The level of significance of the infection with sex, age and season were also tested. Methods: The study was carried out in the FCT, Abuja, Nigeria which is located between latitude 8o and 90 25` North of the equator and longitude 60 45` and 7045` East of the Greenwich meridian. The cross sectional epidemiological method and multi-staged sampling technique were used in this study. Samples were collected from the freshly voided faeces of both apparently healthy and diarrhoeic cattle in selected abattoirs and cattle herds. Enriched samples were analyzed bacteriologically and biochemically after which they were characterised using commercially prepared latex agglutination test kits. Results: A total of 718 faecal samples from cattle were analyzed for the presence of VTEC non-O157. Thirty eight (5.23%) were positive for non-O157. There was no significant association (p > 0.05) between sex and infection with non-O157 VTEC in cattle. There was a significant association (P < 0.05) between age and infection with non-O157 VTEC in cattle. Calves were more associated than the adults. There was also a significant association (P < 0.05) between season and infection with non-O157 VTEC in cattle. The dry season was more associated than the wet season. Conclusion: The study established the occurrence and prevalence of non-O157 VTEC in cattle in FCT, Abuja, Nigeria. As a major food animal in Nigeria, infection in cattle provides an epidemiological causal association to the infection in humans. The result showed that warmer seasons (dry season) stimulate the presence of VTEC infection in animals and thus, as a consequence, increases the number of human cases. The prevalence was also higher in younger calves (< 6 months) probably as a result of undeveloped immune system.

Keywords: prevalence, distribution, Verocytotoxigenic escherichia coli (VTEC), non-O157 serotypes, cattle

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310 Effect of Oxidative Stress on Glutathione Reductase Activity of Escherichia coli Clinical Isolates from Patients with Urinary Tract Infection

Authors: Fariha Akhter Chowdhury, Sabrina Mahboob, Anamika Saha, Afrin Jahan, Mohammad Nurul Islam

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Urinary tract infection (UTI) is frequently experienced by the female population where the prevalence increases with aging. Escherichia coli, one of the most common UTI causing organisms, retains glutathione defense mechanism that aids the organism to withstand the harsh physiological environment of urinary tract, host oxidative immune response and even to affect antibiotic-mediated cell death and the emergence of resistance. In this study, we aimed to investigate the glutathione reductase activity of uropathogenic E. coli (UPEC) by observing the reduced glutathione (GSH) level alteration under stressful condition. Urine samples of 58 patients with UTI were collected. Upon isolation and identification, 88% of the samples presented E. coli as UTI causing organism among which randomly selected isolates (n=9), obtained from urine samples of female patients, were considered for this study. E. coli isolates were grown under normal and stressful conditions where H₂O₂ was used as the stress-inducing agent. GSH level estimation of the isolates in both conditions was carried out based on the colorimetric measurement of 5,5'-dithio-bis (2-nitrobenzoic acid) (DTNB) and GSH reaction product using microplate reader assay. The GSH level of isolated E. coli sampled from adult patients decreased under stress compared to normal condition (p = 0.011). On the other hand, GSH production increased markedly in samples that were collected from elderly subjects (p = 0.024). A significant partial correlation between age and change of GSH level was found as well (p = 0.007). This study may help to reveal ways for better understanding of E. coli pathogenesis of UTI prevalence in elderly patients.

Keywords: Escherichia coli, glutathione reductase activity, oxidative stress, reduced glutathione (GSH), urinary tract infection (UTI)

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309 Microfiber Release During Laundry Under Different Rinsing Parameters

Authors: Fulya Asena Uluç, Ehsan Tuzcuoğlu, Songül Bayraktar, Burak Koca, Alper Gürarslan

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Microplastics are contaminants that are widely distributed in the environment with a detrimental ecological effect. Besides this, recent research has proved the existence of microplastics in human blood and organs. Microplastics in the environment can be divided into two main categories: primary and secondary microplastics. Primary microplastics are plastics that are released into the environment as microscopic particles. On the other hand, secondary microplastics are the smaller particles that are shed as a result of the consumption of synthetic materials in textile products as well as other products. Textiles are the main source of microplastic contamination in aquatic ecosystems. Laundry of synthetic textiles (34.8%) accounts for an average annual discharge of 3.2 million tons of primary microplastics into the environment. Recently, microfiber shedding from laundry research has gained traction. However, no comprehensive study was conducted from the standpoint of rinsing parameters during laundry to analyze microfiber shedding. The purpose of the present study is to quantify microfiber shedding from fabric under different rinsing conditions and determine the effective rinsing parameters on microfiber release in a laundry environment. In this regard, a parametric study is carried out to investigate the key factors affecting the microfiber release from a front-load washing machine. These parameters are the amount of water used during the rinsing step and the spinning speed at the end of the washing cycle. Minitab statistical program is used to create a design of the experiment (DOE) and analyze the experimental results. Tests are repeated twice and besides the controlled parameters, other washing parameters are kept constant in the washing algorithm. At the end of each cycle, released microfibers are collected via a custom-made filtration system and weighted with precision balance. The results showed that by increasing the water amount during the rinsing step, the amount of microplastic released from the washing machine increased drastically. Also, the parametric study revealed that increasing the spinning speed results in an increase in the microfiber release from textiles.

Keywords: front load, laundry, microfiber, microfiber release, microfiber shedding, microplastic, pollution, rinsing parameters, sustainability, washing parameters, washing machine

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308 The Role of Chemokine Family, CXCL-10 Urine as a Marker Diagnosis of Active Lung Tuberculosis in HIV/AIDS Patients

Authors: Dwitya Elvira, Raveinal Masri, Rohayat Bilmahdi

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Human Immunodeficiency Virus (HIV) pandemic increased significantly worldwide. The rise in cases of HIV/AIDS was also followed by an increase in the incidence of opportunistic infection, with tuberculosis being the most opportunistic infection found in HIV/AIDS and the main cause of mortality in HIV/AIDS patients. Diagnosis of tuberculosis in HIV/AIDS patients is often difficult because of the uncommon symptom in HIV/AIDS patients compared to those without the disease. Thus, diagnostic tools are required that are more effective and efficient to diagnose tuberculosis in HIV/AIDS. CXCL-10/IP-10 is a chemokine that binds to the CXCR3 receptor found in HIV/AIDS patients with a weakened immune system. Tuberculosis infection in HIV/AIDS activates chemokine IP-10 in urine, which is used as a marker for diagnosis of infection. The aim of this study was to prove whether IP-10 urine can be a biomarker diagnosis of active lung tuberculosis in HIV-AIDS patients. Design of this study is a cross sectional study involving HIV/AIDS patients with lung tuberculosis as the subject of this study. Forty-seven HIV/AIDS patients with tuberculosis based on clinical and biochemical laboratory were asked to collect urine samples and IP-10/CXCL-10 urine being measured using ELISA method with 18 healthy human urine samples as control. Forty-seven patients diagnosed as HIV/AIDS were included as a subject of this study. HIV/AIDS were more common in male than in women with the percentage in male 85.1% vs. 14.5% of women. In this study, most diagnosed patients were aged 31-40 years old, followed by those 21-30 years, and > 40 years old, with one case diagnosed at age less than 20 years of age. From the result of the urine IP-10 using ELISA method, there was significant increase of the mean value of IP-10 urine in patients with TB-HIV/AIDS co-infection compared to the healthy control with mean 61.05 pg/mL ± 78.01 pg/mL vs. mean 17.2 pg/mL. Based on this research, there was significant increase of urine IP-10/CXCL-10 in active lung tuberculosis with HIV/AIDS compared to the healthy control. From this finding, it is necessary to conduct further research into whether urine IP-10/CXCL-10 plays a significant role in TB-HIV/AIDS co-infection, which can also be used as a biomarker in the early diagnosis of TB-HIV.

Keywords: chemokine, HIV/AIDS, IP-10 urine, tuberculosis

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307 Anatomical and Histological Analysis of Salpinx and Ovary in Anatolian Wild Goat (Capra aegagrus aegagrus)

Authors: Gulseren Kirbas, Mushap Kuru, Buket Bakir, Ebru Karadag Sari

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Capra (mountain goat) is a genus comprising nine species. The domestic goat (C. aegagrus hircus) is a subspecies of the wild goat that is domesticated. This study aimed to determine the anatomical structure of the salpinx and ovary of the Anatolian wild goat (C. aegagrus aegagrus). Animals that were taken to the Kafkas University Wildlife Rescue and Rehabilitation Center, Kars, Turkey, because of various reasons, such as traffic accidents and firearm injuries, were used in this study. The salpinges and ovaries of four wild goats of similar ages, which could not be rescued by the Center despite all interventions, were dissected. Measurements were taken from the right-left salpinx and ovary using digital calipers. The weights of each ovary and salpinx were measured using a precision scale (min: 0.0001 g − max: 220 g, code: XB220A; Precisa, Swiss). The histological structure of the tissues was examined after weighing the organs. The tissue samples were fixed in 10% formaldehyde for 24 h. Then a routine procedure was applied, and the tissues were embedded in paraffin. Mallory’s modified triple staining was used to demonstrate the general structure of the salpinx. The salpinx was found to consist of three different regions (infundibulum, ampulla, and isthmus). These regions consisted of tunica mucosa, tunica muscularis, and tunica serosa. The prismatic epithelial cells were observed in the lamina epithelialis of tunica mucosa in every region, but the prismatic fimbrae cells occurred most in the infundibulum. The ampulla was distinguished by its many mucosal folds. It was the longest region of the salpinx and was joined to the isthmus via the ampullary–isthmus junction. Isthmus was the caudal end of the salpinx joined to the uterus and had the thickest tunica muscularis compared with the other regions. The mean length of the ovary was 13.22 ± 1.27 mm, width was 8.46 ± 0.88 mm, the thickness was 5.67 ± 0.79 mm, and weight was 0.59 ± 0.17 g. The average length of the salpinx was 58.11 ± 14.02 mm, width was 0.80 ± 0.22 mm, the thickness was 0.41 ± 0.01 mm, and weight was 0.30 ± 0.08 g. In conclusion, the Anatolian wild goat, which is included in wildlife diversity in Turkey, has been disappearing due to illegal and uncontrolled hunting as well as traffic accidents in recent years. These findings are believed to contribute to the literature.

Keywords: Anatolian wild goat, anatomy, ovary, salpinx

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306 Hospital Acquired Bloodstream Infections Among Patients With Hematological and Solid Malignancies: Epidemiology, Causative Pathogens and Mortality

Authors: Marah El-Beeli, Abdullah Balkhair, Zakaryia Al Muharmi, Samir Al Adawi, Mansoor Al-Jabri, Abdullah Al Rawahi, Hazaa Al Yahyae, Eman Al Balushi, Yahya M. Al-Farsi

Abstract:

The health care service and the anticancer chemotherapeutics has changed the natural history of cancer into manageable chronic disease and improve the cancer patient’s lifestyle and increase the survival time. Despite that, still, infection is the major dilemma opposing the cancer patient either because of the clinical presentation of the cancer type and impaired immune system or as a consequence of anticancer therapy. This study has been conducted to1) track changes in the epidemiology of hospital-acquired bloodstream infections among patients with malignancies in the last five years. 2) To explore the causative pathogens and 3) the outcome of HA-BSIs in patients with a different types of malignancies. An ampi-directional study (retrospective and prospective follow up) of patients with malignancies admitted at Sultan Qaboos University hospital (570-bed tertiary hospital) during the study period (from January 2015 to December 2019). The cumulative frequency and prevalence rates of HA-BSIs by patients and isolates were calculated. In addition, the cumulative frequency of participants with single versus mixed infections and types of causative micro-organisms of HA-BSIs were obtained. A total of 1246 event of HA-BSIs has occurred during the study period. Nearly the third (30.25%) of the HA-BSI events was identified among 288 patients with malignancies. About 20% of cases were mixed infections (more than one isolate). Staphylococcus spp were the predominant isolated pathogen (24.7%), followed by Klebsiella spp (15.8%), Escherichia spp (13%), and Pseudomonas spp (9.3%). About half (51%) of cases died in the same year, and (64%) of the deaths occur within two weeks after the infection. According to the observations, no changes in the trends of epidemiology, causative pathogens, morbidity, and mortality rates in the last five years.

Keywords: epidemiology, haematological malignancies, hospital acquired bloodstream infections, solid malignancies

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305 LTF Expression Profiling Which is Essential for Cancer Cell Proliferation and Metastasis, Correlating with Clinical Features, as Well as Early Stages of Breast Cancer

Authors: Azar Heidarizadi, Mahdieh Salimi, Hossein Mozdarani

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Introduction: As a complex disease, breast cancer results from several genetic and epigenetic changes. Lactoferrin, a member of the transferrin family, is reported to have a number of biological functions, including DNA synthesis, immune responses, iron transport, etc., any of which could play a role in tumor progression. The aim of this study was to investigate the bioinformatics data and experimental assay to find the pattern of promoter methylation and gene expression of LTF in breast cancer in order to study its potential role in cancer management. Material and Methods: In order to evaluate the methylation status of the LTF promoter, we studied the MS-PCR and Real-Time PCR on samples from patients with breast cancer and normal cases. 67 patient samples were conducted for this study, including tumoral, plasma, and normal tissue adjacent samples, as well as 30 plasma from normal cases and 10 tissue breast reduction cases. Subsequently, bioinformatics analyses such as cBioPortal databases, string, and genomatix were conducted to disclose the prognostic value of LTF in breast cancer progression. Results: The analysis of LTF expression showed an inverse relationship between the expression level of LTF and the stages of tissues of breast cancer patients (p<0.01). In fact, stages 1 and 2 had a high expression in LTF, while, in stages 3 and 4, a significant reduction was observable (p < 0.0001). LTF expression frequently alters with a decrease in the expression in ER⁺, PR⁺, and HER2⁺ patients (P < 0.01) and an increase in the expression in the TNBC, LN¯, ER¯, and PR- patients (P < 0.001). Also, LTF expression is significantly associated with metastasis and lymph node involvement factors (P < 0.0001). The sensitivity and specificity of LTF were detected, respectively. A negative correlation was detected between the results of level expression and methylation of the LTF promoter. Conclusions: The altered expression of LTF observed in breast cancer patients could be considered as a promotion in cell proliferation and metastasis even in the early stages of cancer.

Keywords: LTF, expression, methylation, breast cancer

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