Search results for: serum proteins

Authors: Mohammad Naeem Lakanwall

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Background:The thyroid gland, located just below the vocal cord on each side of and anterior to the trachea, is one of the main endocrine glands. Its normal weight is 15 to 20 grams in adults. The thyroid secretes two most important hormones, thyroxine and triiodothyronine, usually called T4 and T3, respectively. These hormones greatly increase the metabolic rate of the body. In addition to T3 and T4, the thyroid gland secrets calcitonin as well which is a significant hormone for calcium metabolism. Objective: The aim of this study is to estimate the proportion of and to identify factors associated with thyroid dysfunction among individuals coming to a tertiary care facility in Kabul, Afghanistan. Material and Methods: An analytical cross-sectional study design was conducted from July to Sep 2018. Blood samples were obtained, serum TSH levels were measured, and the patients were divided into three diagnostic categories according to their serum TSH concentrations. 1) Hypothyroidism 2) Hyperthyroidism 3) Normal thyroid Results: A total of 127 individuals were part of the sample for the final analysis. The majority of study participants (77%) were females. A large number of the participants (92%) did not have a family history of thyroid dysfunction and the majority of the female participants, (85%) were not pregnant in the last two years. Furthermore, 98% of participants, were non-smokers. Conclusion: The findings of the current study showed a high prevalence of thyroid dysfunctions in individuals coming to FMIC for thyroid functions tests. The findings also indicated that aging and smoking are the factors associated with thyroid dysfunctions. Further studies are needed to find out the prevalence of and factors associated with thyroid dysfunctions.

Keywords: Afghanistan, Kabul, hypothyroidism, hyperthyroidism, triiodothyronine, thyroxine

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Authors: Zoe Goldberger, Priscilla S. Briquez, Jeffrey A. Hubbell

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Tumor cells have mutations resulting from genetic instability that the immune system can actively recognize. Immune checkpoint immunotherapy (ICI) is commonly used in the clinic to re-activate immune reactions against mutated proteins, called neoantigens, resulting in tumor remission in cancer patients. However, only around 20% of patients show durable response to ICI. While tumor mutational burden (TMB) has been approved by the Food and Drug Administration (FDA) as a criterion for ICI therapy, the relevance of the subcellular localizations of the mutated proteins within the tumor cell has not been investigated. Here, we hypothesized that localization of mutations impacts the effect of immune responsiveness to ICI. We analyzed publicly available tumor mutation sequencing data of ICI treated patients from 3 independent datasets. We extracted the subcellular localization from the UniProtKB/Swiss-Prot database and quantified the proportion of membrane, cytoplasmic, nuclear, or secreted mutations per patient. We analyzed this information in relation to response to ICI treatment and overall survival of patients showing with 1722 ICI-treated patients that high mutational burden localized at the membrane (mTMB), correlate with ICI responsiveness, and improved overall survival in multiple cancer types. We anticipate that our results will ameliorate predictability of cancer patient response to ICI with potential implications in clinical guidelines to tailor ICI treatment. This would not only increase patient survival for those receiving ICI, but also patients’ quality of life by reducing the number of patients enduring non-effective ICI treatments.

Keywords: cancer, immunotherapy, membrane neoantigens, efficacy prediction, biomarkers

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Authors: Mohammed Sabbah, Prospero Di Pierro, Raffaele Porta

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Protein-based edible films and coatings have attracted an increasing interest in recent years since they might be used to protect pharmaceuticals or improve the shelf life of different food products. Among them, several plant proteins represent an abundant, inexpensive and renewable raw source. These natural biopolymers are used as film forming agents, being able to form intermolecular linkages by various interactions. However, without the addition of a plasticizing agent, many biomaterials are brittle and, consequently, very difficult to be manipulated. Plasticizers are generally small and non-volatile organic additives used to increase film extensibility and reduce its crystallinity, brittleness and water vapor permeability. Plasticizers normally act by decreasing the intermolecular forces along the polymer chains, thus reducing the relative number of polymer-polymer contacts, producing a decrease in cohesion and tensile strength and thereby increasing film flexibility allowing its deformation without rupture. The most commonly studied plasticizers are polyols, like glycerol (GLY) and some mono or oligosaccharides. In particular, GLY not only increases film extensibility but also migrates inside the film network often causing the loss of desirable mechanical properties of the material. Therefore, replacing GLY with a different plasticizer might help to improve film characteristics allowing potential industrial applications. To improve film properties, it seemed of interest to test as plasticizers some cationic small molecules like polyamines (PAs). Putrescine, spermidine (SPD), and spermine are PAs widely distributed in nature and of particular interest for their biological activities that may have some beneficial health effects. Since PAs contains amino instead of hydroxyl functional groups, they are able to trigger ionic interactions with negatively charged proteins. Bitter vetch (Vicia ervilia; BV) is an ancient grain legume crop, originated in the Mediterranean region, which can be found today in many countries around the world. This annual Vicia genus shows several favorable features, being their seeds a cheap and abundant protein source. The main objectives of this study were to investigate the effect of different concentrations of SPD on the mechanical and permeability properties of films prepared with native or heat denatured BV proteins in the presence of different concentrations of SPD and/or GLY. Therefore, a BV seed protein concentrate (BVPC), containing about 77% proteins, was used to prepare film forming solutions (FFSs), whereas GLY and SPD were added as film plasticizers, either singly or in combination, at various concentrations. Since a primary plasticizer is generally defined as a molecule that when added to a material makes it softer, more flexible and easier to be processed, our findings lead to consider SPD as a possible primary plasticizer of protein-based films. In fact, the addition of millimolar concentrations of SPD to BVPC FFS allowed obtaining handleable biomaterials with improved properties. Moreover, SPD can be also considered as a secondary plasticizer, namely an 'extender', because of its ability even to enhance the plasticizing performance of GLY. In conclusion, our studies indicate that innovative edible protein-based films and coatings can be obtained by using PAs as new plasticizers.

Keywords: edible films, glycerol, plasticizers, polyamines, spermidine

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Authors: Alena Semeradtova, Marcel Stofik, Lucie Mareckova, Petr Maly, Ondrej Stanek, Jan Maly

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Recently, novel strategies based on so-called molecular evolution were shown to be effective for the production of various peptide ligand libraries with high affinities to molecular targets of interest comparable or even better than monoclonal antibodies. The major advantage of these peptide scaffolds is mainly their prevailing low molecular weight and simple structure. This study describes a new high-affinity binding molecules based immunesensor using a simple optical system for human serum albumin (HSA) detection as a model molecule. We present a comparison of two variants of recombinant binders based on albumin binding domain of the protein G (ABD) performed on micropatterned glass chip. Binding domains may be tailored to any specific target of interest by molecular evolution. Micropatterened glass chips were prepared using UV-photolithography on chromium sputtered glasses. Glass surface was modified by (3-aminopropyl)trietoxysilane and biotin-PEG-acid using EDC/NHS chemistry. Two variants of high-affinity binding molecules were used to detect target molecule. Firstly, a variant is based on ABD domain fused with TolA chain. This molecule is in vivo biotinylated and each molecule contains one molecule of biotin and one ABD domain. Secondly, the variant is ABD domain based on streptavidin molecule and contains four gaps for biotin and four ABD domains. These high-affinity molecules were immobilized to the chip surface via biotin-streptavidin chemistry. To eliminate nonspecific binding 1% bovine serum albumin (BSA) or 6% fetal bovine serum (FBS) were used in every step. For both variants range of measured concentrations of fluorescently labelled HSA was 0 – 30 µg/ml. As a control, we performed a simultaneous assay without high-affinity binding molecules. Fluorescent signal was measured using inverse fluorescent microscope Olympus IX 70 with COOL LED pE 4000 as a light source, related filters, and camera Retiga 2000R as a detector. The fluorescent signal from non-modified areas was substracted from the signal of the fluorescent areas. Results were presented in graphs showing the dependence of measured grayscale value on the log-scale of HSA concentration. For the TolA variant the limit of detection (LOD) of the optical immunosensor proposed in this study is calculated to be 0,20 µg/ml for HSA detection in 1% BSA and 0,24 µg/ml in 6% FBS. In the case of streptavidin-based molecule, it was 0,04 µg/ml and 0,07 µg/ml respectively. The dynamical range of the immunosensor was possible to estimate just in the case of TolA variant and it was calculated to be 0,49 – 3,75 µg/ml and 0,73-1,88 µg/ml respectively. In the case of the streptavidin-based the variant we didn´t reach the surface saturation even with the 480 ug/ml concentration and the upper value of dynamical range was not estimated. Lower value was calculated to be 0,14 µg/ml and 0,17 µg/ml respectively. Based on the obtained results, it´s clear that both variants are useful for creating the bio-recognizing layer on immunosensors. For this particular system, it is obvious that the variant based on streptavidin molecule is more useful for biosensing on glass planar surfaces. Immunosensors based on this variant would exhibit better limit of detection and wide dynamical range.

Keywords: high affinity binding molecules, human serum albumin, optical immunosensor, protein G, UV-photolitography

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Authors: Oyegunle Emmanuel Oke, A. O. Onabajo, M. O. Abioja, F. O. Sorungbe, D. E. Oyetunji, J. A. Abiona, A. O. Ladokun, O. M. Onagbesan

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An experiment was conducted to determine the effect of different dietary inclusion levels of Moringa oleifera leaf powder (MOLP) on growth and physiological responses of broiler chickens during hot-dry season in Nigeria. Two hundred and forty (240) day-old commercial broiler chicks were randomly allotted to four dietary treatments having four replicates each. Each replicate had 15 birds. The levels of inclusion were 0g (Control group), 4g, 8g and 12g/Kg feed. The experiment lasted for eight weeks. The results of the study revealed that the initial body weight was significantly (P < 0.05) higher in birds fed 12g/kg diet than those fed 0, 4, and 8g MOLP. The birds fed 0, 4 and 8g/kg diet however had similar weights. The final body weight was significantly (P < 0.05) higher in the birds fed 12g MOLP than those fed 0, 4 and 8g MOLP. The final weights were similar in the birds fed 4 and 8g/kg diet but higher (P < 0.05) than those of the birds in the control group. The body weight gain was similar in birds fed 0 and 4g MOLP but significantly higher (P < 0.05) than those of the birds in 12g/kg diet. There were no significant differences (P > 0.05) in the feed intake. The serum albumin of the birds fed 12g MOLP/Kg diet (48.85g/L) was significantly (P < 0.05) higher than the mean value of those fed the control diet 0 and 8g MOLP/Kg diets having 36.05 and 37.10g/L respectively. Birds fed 12g MOLP/Kg feed recorded the lowest level of triglyceride (122.75g/L) which was significantly (P < 0.05) lower than those of the birds fed 0 and 4g/kg diet MOLP. The serum corticosterone decreased with increase in MOLP inclusion levels. The birds fed 12g MOLP had the least value. This study has shown that MOLP may contain potent antioxidants capable of ameliorating the effects of heat stress in broiler chickens with 12g MOLP inclusion.

Keywords: physiology, performance, heat stress, anti-oxidant

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Authors: Ahlem Bahi, Youcef Necib, Sakina Zerizer, Cherif Abdennour, Mohamed Salah Boulakoud

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Mercury (II) is a highly toxic metal which induces oxidative stress in the body. In this study, we aimed to investigate the possible protective effect of olive oil, an antioxidant agent, against experimental mercury toxicity in rat model. Administration of mercuric chloride induced significant increase in serum: ALT, AST, and LPA activities; interleukine1, interleukine6, tumor necrosis factor α (TNFα), creatinine, urea, and uric acid levels. Mercuric chloride also induced oxidative stress, as indicate by decreased tissue of GSH level, GSH-Px, and GST activities along with increase the level of lipid peroxidation. Furthermore, treatment with mercuric chloride caused a marked elevation of kidney and liver weight and decreased body weight. Virgin olive oil treatment markedly reduced elevated serum: AST, ALT, and LPA activities; interleukine1, interleukine6, tumor necrosis factor α (TNFα), creatinine, urea, and uric acid levels and contracted the deterious effects of mercuric chloride on oxidative stress markers changes caused by HgCl2 in tissue as compared to control group. Our results implicate that mercury induced oxidative damage in liver and kidney tissue protected by virgin olive oil, with its antioxidant effects.

Keywords: mercury, antioxidant enzymes, pro-inflammatory cytokine, virgin olive oil, lipid peroxidation

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Authors: Hafiza Javaria Ashraf, Xinghong Wang, Zhanghong Shi, Youming Hou

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Insect’s innate immunity depends on a variety of defense responses for the recognition of invading pathogens. Pathogen recognition involves particular proteins known as pattern recognition receptors (PRRs). These PRRs interact with pathogen-associated molecular patterns (PAMPs) present on the surface of pathogens to distinguish between self and non-self. C-type lectins (CTLs) belong to a superfamily of PPRs which involved in insect immunity and defense mechanism. Rhynchophorus ferrugineus Olivier is a devastating pest of Palm cultivations in China. Although studies on R. ferrugineus immune mechanism and host defense have conducted, however, the role of CTL in immune responses of R. ferrugineus remains elusive. Here, we report RfCTL, which is a secreted protein containing a single-CRD domain. The open reading frame (ORF) of CTL is 226 bp, which encodes a putative protein of 168 amino acids. Transcript expression analysis revealed that RfCTL highly expressed in immune-related tissues, i.e., hemolymph and fat body. The abundance of RfCTL in the gut and fat body dramatically increased upon Staphylococcus aureus and Escherichia coli bacterial challenges, suggesting a role in defense against gram-positive and gram-negative bacterial infection. Taken together, we inferred that RfCTL might be involved in the immune defense of R. ferrugineus and established a solid foundation for future studies on R. ferrugineus CTL domain proteins for better understanding of insect immunity.

Keywords: biological invasion, c-type lectin, insect immunity, Rhynchophorus ferrugineus Oliver

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Authors: Ahmed Alkarimi, Kevin Welham

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Organic mixed mode monolithic columns were fabricated from; glycidyl methacrylate-co-ethylene dimethacrylate-co-stearyl methacrylate, using glycidyl methacrylate and stearyl methacrylate as co monomers representing 30% and 70% respectively of the liquid volume with ethylene dimethacrylate crosslinker and 2,2-dimethoxy-2-phenylacetophenone as the free radical initiator. The monomers were mixed with a binary porogenic solvent, comprising propan-1-ol, and methanol (0.825 mL each). The monolith was formed by photo polymerization (365 nm) inside a borosilicate glass tube (1.5 mm ID and 3 mm OD x 50 mm length). The monolith was observed to have formed correctly by optical examination and generated reasonable backpressure, approximately 650 psi at a flow rate of 0.2 mL min⁻¹ 50:50 acetonitrile: water. The morphological properties of the monolithic columns were investigated using scanning electron microscopy images, and Brunauer-Emmett-Teller analysis, the results showed that the monolith was formed properly with 19.98 ± 0.01 mm² surface area, 0.0205 ± 0.01 cm³ g⁻¹ pore volume and 6.93 ± 0.01 nm average pore size. The polymer monolith formed was further investigated using proton nuclear magnetic resonance, and Fourier transform infrared spectroscopy. The monolithic columns were investigated using high-performance liquid chromatography to test their ability to separate different samples with a range of properties. The columns displayed both hydrophobic/hydrophilic and hydrophobic/ion exchange interactions with the compounds tested indicating that true mixed mode separations. The mixed mode monolithic columns exhibited significant separation of proteins.

Keywords: LC separation, proteins separation, monolithic column, mixed mode

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Authors: O. Soji-Omoniwa, N. O. Muhammad, L. A. Usman, B. P. Omoniwa

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This study was conducted to investigate the effect of the leaf essential oil of C. sinensis harvested at 7.00a.m and 4.00p.m on some Liver and Kidney function indices of diabetic rats as well as investigate the effect of time of harvest on the observed effect. Experimental animals were divided into 4 groups (A, B, C and D). Diabetes mellitus was induced in all animals, except the normal control group (Group A), by injecting 150mg/kg body weight of alloxan monohydrate intraperitoneally. Group A received distilled water while group B (diabetic control group) was not treated. Group C and D were treated with leaf essential oil of C. sinensis harvested at 7.00 a.m and 4.00 p.m respectively at a dose of 110 mg/kg body weight every other day for 15 days. Alkaline phosphatase (ALP), Alanine Transaminase (ALT) and Aspartate Transaminase (AST) activity was evaluated in the serum, Liver and Kidney of studied animals. Total and Direct Bilirubin level, Total Protein and Globulin, Creatinine and Urea level were also evaluated. Result showed that creatinine and urea, serum ALP, AST and ALT levels was significantly reduced (p < 0.05), while the levels of total Protein and Globulin increased significantly (p < 0.05) for the treated animals compared to the diabetic control group. In conclusion, the leaf essential oil of Citrus sinensis ameliorated the impaired renal and liver function; however, the time of harvest of the leaf does not significantly affect its ameliorative effect.

Keywords: C. sinensis, function indices, harvest time, leaf essential oil.

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Authors: Mehrnaz Mostafavi

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The current research aims to explore a method for creating nano-ring structures through chemical reduction. By employing a direct reduction process at a controlled, slow pace, and concurrently introducing specific reduction agents, the goal is to fabricate these unique nano-ring formations. The deliberate slow reduction of nanoparticles within this process helps prevent spatial hindrances caused by the reduction agents. The timing of the reduction of metal atoms, facilitated by these agents, emerges as a crucial factor influencing the creation of nano-ring structures. In investigation involves a chemical approach utilizing bovine serum albumin and human serum albumin as organic reducing agents to produce gold nano-rings. The controlled reduction of metal atoms at a slow pace and under specific pH conditions plays a pivotal role in the successful fabrication of these nanostructures. Optical spectroscopic analyses revealed distinctive plasmonic behavior in both visible and infrared spectra, owing to the collective movement of electrons along the inner and outer walls of the gold nano-rings. Importantly, these ring-shaped nanoparticles exhibit customizable plasmon resonances in the near-infrared spectrum, a characteristic absent in solid particles of similar sizes. This unique attribute makes the generated samples valuable for applications in Nanomedicine and Nanobiotechnology, leveraging the distinct optical properties of these nanostructures.

Keywords: nano-ring structure, nano-particles, reductant agents, plasmon resonace

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Authors: T. N. Girish, B. E. Pradeep, Ravi Parkash

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Insects from tropical wet or dry seasons are likely to cope starvation stress through seasonal phenotypic plasticity in energy metabolites. Accordingly, we analyzed such plastic changes in Zaprionus indianus flies reared under wet or dry season-specific conditions; and also after adult acclimation at 32℃ for 1 to 6 days; and to low (40% RH) or high (70% RH) humidity. Both thermal or humidity acclimation revealed significant accumulation of body lipids for wet season flies but low humidity acclimation did not change the level of body lipids in dry season flies. Developmental and adult acclimation showed sex specific differences i.e., starvation resistance and body lipids were higher in the males of dry season but in females of wet season. We found seasonal and sex specific differences in the relative level for body lipids at death; and in the rates of accumulation or utilization of energy metabolites (body lipids, carbohydrates and proteins). Body lipids constitute the preferred energy source under starvation for flies of both the seasons. However, utilization of carbohydrates (~20% to 30%) and proteins (~20% to 25%) was evident only in dry season flies. Higher starvation resistance after thermal or humidity acclimation is achieved by increased accumulation of lipids. Adult acclimation of wet or dry season flies revealed plastic changes in mean daily fecundity despite reduction in fecundity under starvation. Thus, thermal or humidity induced plastic responses in body lipids support starvation resistance under wet or dry seasons.

Keywords: heat or humidity acclimation, plastic changes in body lipids and starvation resistance, tropical drosophilid, Wet- or Dry seasons, Zaprionus indianus

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Authors: Mohammad Naeem Lakanwall, Jamshid Abdul-Ghafar

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Background: Endocrinopathies are a commonly occurring entity, particularly those of the thyroid gland; however, there is a lack of scientific literature from Afghanistan, a country with very limited health care facilities and resources. To our best knowledge, this is the first study aimed to describe the frequency of occurrence and factors associated with thyroid dysfunction in the Afghan population. The aim of this study is to estimate the frequency and to identify factors associated with thyroid dysfunction among individuals coming to a tertiary care facility in Kabul, Afghanistan. Methods: A cross-sectional study was conducted from July to Sep 2018 at the Department of Clinical Pathology, French Medical Institute for Mothers and Children (FMIC), Kabul, Afghanistan. Blood samples were obtained, serum TSH levels were analyzed, and the patients were divided into three diagnostic categories according to their serum TSH concentrations: 1) hypothyroidism, 2) hyperthyroidism, 3) normal. Results: A total of 127 individuals were included in the final analysis. The majority of study participants (77%) were females. A large number of the participants (92%) did not have a family history of thyroid dysfunction. 74% of the participants in the study had normal TSH levels classified as normal thyroid function, (14%) had lower TSH levels, and (12%) higher TSH levels, classified as hyper and hypothyroid, respectively. Conclusions: The findings of the current study showed a high frequency of thyroid dysfunctions from a single center. Further large-scale studies are needed to find out the prevalence and document this entity for better health outcomes in the country.

Keywords: Afghanistan, factors, frequency, hypothyroid, hyperthyroid, thyroid, thyroid stimulating hormone

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

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

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

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Authors: Hala Demerdash, Ola M. Zanaty, Emad Eldin Arida

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Background: This study investigated the micoRNA expression changes induced by Sevoflurane and Propofol and their effects on liver functions. Patients and methods: The study was designed as randomized controlled study, carried out on 200 adult patients, scheduled for major surgeries under general anesthesia (GA). Patients were randomly divided into four groups; groups SC and PC included chronic hepatitis C (CHC) patients where SC group are patients receiving Sevoflurane, and PC group are patients receiving Propofol anesthesia. While S and P groups included non- hepatitis patients; S group are patients receiving Sevoflurane and P group are patients receiving Propofol. Anesthesia in Group S and SC patients was maintained by sevoflurane, while anesthesia in Group P and PC patients was maintained by propofol infusion. Blood samples were analyzed for PT, PTT and liver enzymes. Serum samples were analyzed for microRNA before and after surgery. Results: Results show miRNA-122 and miRNA-21 were absent in serum of S and P groups in pre-operative samples. However, they were expressed in SC and PC groups. In post-operative samples; miRNA-122 revealed an increased expression in all groups; with more exaggerated response in SC group. On the other hand miRNA-21 revealed increased expression in both SC and PC groups; a slight expression in S group with absent expression in P group. There was a post-operative negative correlation between miR-122 and ALT (r=-0.46) in SC group and (r=-0.411) in PC group and positive correlation between ALT and miR-21 (r=0.335) in SC group and (r=0.379) in PC group. The amount of blood loss was positively correlated with miR-122 (r=0.366) in SC group and (r=0.384) in PC group. Conclusion: Propofol anesthesia is safer than Sevoflurane anesthesia in patients with CHC. Sevoflurane and Propofol anesthesia affect miRNA expression in both CHC and non-hepatitis patients.

Keywords: anesthesia, chronic hepatitis C, micoRNA, propofol, sevoflurane

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Authors: Pornrut Rabintossaporn, Suphaket Saenthaweesuk, Amornnat Thuppia, Nuntiya Somparn

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Several dietary and herbal plants have been shown to possess cytoprotective and antioxidant effects with various mechanisms of action. The aim of this study was to determine the antioxidant effects and its mechanism of aqueous leaves extract of Ocimum americanum (OA), commonly known as American basil or 'hoary basil', in rat. The extract was screened for its phytochemical contents and antioxidant activity in vitro. Moreover, the extract was studied in rats to evaluate its effects in vivo. Rats were orally administered with the extract at the dose of 100, 200 and 400 mg/kg for 28 days. Phytochemical screening of plant extracts revealed the presence of alkaloid, cardiac glycosides, tannin and steroid compounds. The extract contained phenolic compounds 36.91 ± 0.66 mg of gallic acid equivalents per gram OA extract. The free radical scavenging activity assessed by DPPH assay gave IC50 of 41.27 ± 1.86 µg/mL, which is relatively lower than that of BHT with IC50 of 12.34 ± 1.14µg/mL. In the animals, the extract was well tolerated by the animals throughout the 28 days of study as shown by normal serum levels AST, ALP, ALT, BUN and Cr as well as normal histology of liver and pancreatic and kidney tissue. The protein expression of antioxidant enzymes, γ-glutamylcysteine ligase (γ-GCL) in liver was significantly increased compared with normal control. Consistent with the induction of γ-GCL protein expression significantly reduction of serum oxidative stress marker malondialdehyde (MDA) was found in rat treated with OA extract compared with control. Taken together, this study provides evidence that Ocimum americanum exhibits direct antioxidant properties and can induce cytoprotective enzyme in vivo.

Keywords: antioxidant, γ-glutamylcysteine ligase, MDA, Ocimum americanum

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Authors: Emma Plante, Charles Ekwunwa, Diego Illanes

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Introduction: Serum Sickness-Like Reaction (SSLR) is an inflammatory immune response characterized by a rash, polyarthralgias, and fever. SSLR usually occurs in response to a new medication (most commonly antibiotics, anticonvulsants, or antiinflammatory agents) and is believed to involve the formation of drug-specific immune complexes. Here we present a case of a 16-year-old female patient who developed an SSLR in response to the D-mannose-containing over-the-counter supplement, Uqora, used to promote bladder health. Methodology: The methodology for this study included a thorough literature search for other cases of SSLR associated with D-Mannose containing products. Data collection was performed through a review of the patient’s medical record, including history, physical examination, relevant laboratory results, and treatment plan. Findings: A 16-year-old female with a history of overactive bladder and anemia presented with a diffuse urticarial rash, headaches, joint pain, and swelling for three days. Her medications included oral contraceptive pills, iron, mirabegron, UQora, and a probiotic. Physical examination revealed a diffuse urticarial rash, and her musculoskeletal exam revealed swelling and tenderness in her wrists. Her CBC, basic metabolic panel, liver function panel, lyme titers, and urinalysis were all within normal limits. The patient was referred to an allergist, who diagnosed her with SSLR. All medications were discontinued, and she was treated with a 7-day course of prednisone and cetirizine. Her symptoms resolved, and her medications were slowly resumed sequentially over several months. However, UQora triggered a recurrence of her symptoms, and it was identified as the culprit medication. Consequently, UQora was permanently discontinued, and the patient has remained symptom-free. Conclusion: This case report describes the first documented case of SSLR caused by UQora (active ingredient D-mannose). D-Mannose is a monosaccharide found in many plants and fruits, and it is commonly used to prevent urinary tract infections. While the clinical features and timeline, in this case, were typical of SSLR, UQora as the trigger was highly unusual. Clinicians should be aware of the diverse triggers of SSLR and the importance of prompt identification and management to enhance patient safety. It is possible D-mannose was not the trigger, and further research is necessary to better understand the potential therapeutic applications of D-mannose, as well as the potential risks and interactions.

Keywords: serum sickness-like reaction, d-mannose, hypersensitivity reaction, urticaria

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Authors: Shekher Kummari, V. Sunil Kumar, K. Vengatajalabathy Gobi

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A simple, cost-effective, reusable and reagent-free electrochemical biosensor is developed with functionalized multiwall carbon nanotube paste electrode (f-CNTPE) for the sensitive and selective determination of the important chemotherapeutic drug methotrexate (MTX), which is widely used for the treatment of various cancer and autoimmune diseases. The electrochemical response of the fabricated electrode towards the detection of MTX is examined by cyclic voltammetry (CV), differential pulse voltammetry (DPV) and square wave voltammetry (SWV). CV studies have shown that f-CNTPE electrode system exhibited an excellent electrocatalytic activity towards the oxidation of MTX in phosphate buffer (0.2 M) compared with a conventional carbon paste electrode (CPE). The oxidation peak current is enhanced by nearly two times in magnitude. Applying the DPV method under optimized conditions, a linear calibration plot is achieved over a wide range of concentration from 4.0×10⁻⁷ M to 5.5×10⁻⁶ M with the detection limit 1.6×10⁻⁷ M. further, by applying the SWV method a parabolic calibration plot was achieved starting from a very low concentration of 1.0×10⁻⁸ M, and the sensor could detect as low as 2.9×10⁻⁹ M MTX in 10 s and 10 nM were detected in steady state current-time analysis. The f-CNTPE shows very good selectivity towards the specific recognition of MTX in the presence of important biological interference. The electrochemical biosensor detects MTX in-vitro directly from pharmaceutical sample, undiluted urine and human blood serum samples at a concentration range 5.0×10⁻⁷ M with good recovery limits.

Keywords: amperometry, electrochemical detection, human blood serum, methotrexate, MWCNT, SWV

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Authors: Mahsa Taghavi

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

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

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Authors: Antonis Elia, Theo Loizou, Gladys Onambele-Pearson, Matthew Barlow, Georgina Stebbings

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Hypoxic conditions have been reported to enhance red blood cell production in both acclimatised low-landers and altitude adapted populations. This process is mediated by the erythropoietin hormone, which is released predominantly by the hypoxic kidney. A higher haemoglobin concentration was previously reported in elite breath-hold divers when compared to elite-skiers and untrained individuals. Therefore, the present study aimed to investigate whether apnoea induced hypoxia could induce a significant increase in serum erythropoietin concentration in recreational breath-hold divers which would provide an explanation to the higher haemoglobin levels observed in elite breath-hold divers. Identifying whether apnoea induced hypoxia induces a significant increase in serum erythropoietin might suggest that apnoea can be used as an alternative acclimatisation method to high altitude exposure. Seven healthy, recreational male breath-hold divers performed two sets of five 180 second breath-holds with a ten-minute supine rest between each set and a two-minute seated rest between each apnoea. During each breath-hold, participant’s heart rate and peripheral oxygen saturation levels were recorded every subsequent 10 seconds until the end of the 180 second breath-hold. After each 180 second breath-hold a capillary blood sample was collected from the finger to identify circulating haemoglobin levels. Following completion of the apnoeic protocol, three blood samples were collected at 30, 90 and 180 minutes to measure circulating erythropoietin levels. A significant interaction between erythropoietin and time was observed (F(3,18)= 4.72, p < 0.001), with significant increases in erythropoietin evident at 30 (t(6)= -5.035, p < 0.0590 (t(6)= -6.162, p < 0.05) and 180 (t(6)= - 7.232, p < 0.001) minutes post the last apnoea when compared to baseline. Corresponding average increases when compared to baseline were 16% at 30, 23% at 90 and 40% at 180 minutes post the last apnoea. A significant interaction between haemoglobin and time was observed (F(78,84)= 20.814, p < 0.001), with significant increases in haemoglobin evident at the fifth (t(29)= -1.124, p < 0.001), ninth (t(29)= -1.357, p < 0.001) and tenth (t(29)= -1.211, p < 0.05) apnoeas when compared to baseline. A significant interaction between peripheral oxygen saturation and time was observed (F(10,60)= 408.23, p < 0.001). The present study demonstrates that a series of ten 180 second breath-holds is sufficient to induce a significant increase in the circulating erythropoietin concentration of recreational breath hold divers. These observations may suggest that apnoea induced hypoxia may be used as an alternative acclimatisation method to high altitude exposure.

Keywords: apnoea, breath-holding, diving reflex, erythropoietin, haemoglobin

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Authors: Ibrahim Aly, Manal Ahmed, Mahmoud M. El-Shall

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Filariasis is a parasitic disease caused by small roundworms. The filarial worms are transmitted and spread by blood-feeding black flies and mosquitoes. Lymphatic filariasis (Elephantiasis) is caused by Wuchereriabancrofti, Brugiamalayi, and Brugiatimori. Elimination of Lymphatic filariasis necessitates an increasing demand for valid, reliable, and rapid diagnostic kits. Nanodiagnostics involve the use of nanotechnology in clinical diagnosis to meet the demands for increased sensitivity, specificity, and early detection in less time. The aim of this study was to evaluate the nano-based enzymelinked immunosorbent assay (ELISA) and novel nano-chip card as a rapid test for detection of filarial antigen in serum samples of human filariasis in comparison with traditional -ELISA. Serum samples were collected from an infected human with filarial gathered across Egypt's governorates. After receiving informed consenta total of 45 blood samples of infected individuals residing in different villages in Gharbea governorate, which isa nonendemic region for bancroftianfilariasis, healthy persons living in nonendemic locations (20 persons), as well as sera from 20 other parasites, affected patients were collected. The microfilaria was checked in thick smears of 20 µl night blood samples collected during 20-22 hrs. All of these individuals underwent the following procedures: history taking, clinical examination, and laboratory investigations, which included examination of blood samples for microfilaria using thick blood film and serological tests for detection of the circulating filarial antigen using polyclonal antibody- ELISA, nano-based ELISA, and nano-chip card. In the present study, a recently reported polyoclonal antibody specific to tegumental filarial antigen was used in developing nano-chip card and nano-ELISA compared to traditional ELISA for the detection of circulating filarial antigen in sera of patients with bancroftianfilariasis. The performance of the ELISA was evaluated using 45 serum samples. The ELISA was positive with sera from microfilaremicbancroftianfilariasis patients (n = 36) with a sensitivity of 80 %. Circulating filarial antigen was detected in 39/45 patients who were positive for circulating filarial antigen using nano-ELISA with a sensitivity of 86.6 %. On the other hand, 42 out of 45 patients were positive for circulating filarial antigen using nano-chip card with a sensitivity of 93.3%.In conclusion, using a novel nano-chip assay could potentially be a promising alternative antigen detection test for bancroftianfilariasis.

Keywords: lymphatic filariasis, nanotechnology, rapid diagnosis, elisa technique

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Authors: K. Shafiei, A. Pirestani, G. Ghalamkari, S. Safavipour

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Use of GnRH or its agonists to increase conception rates should be based on an understanding of GnRH-induced biological effects on the reproductive-endocrine system. This effect may occur through GnRH-stimulated LH surge stimulating production of progesterone by corpus luteum.the aim of this study was to compare the effects on reproductive efficiency of a luteolytic dose of a synthetic prostaglandin Cloprostenol Sodium versus ainjectable progesterone and Luliberin- A on Follicle estrogen and progesterone levels.In this study, we used45 head of holstein dairy heifersin the three treatments, with 15 replicates per treatment were performed in random groups. all the heifers before the projects is began in two steps injection 3 mL CloprostenolSodium with an interval of 11 days been synchronized and 10 days later, second injection of prostaglandin was conducted after that we started below protocol:Control group (daily sodium chloride serum injection 1 cc), Group B: Day Zero, intramuscular injection of 15 mg Luliberin- A + every other day injection of 3 cc progesterone + day 7, injection of Cloprostenol Sodium+ day 9, injection of 15 mg Luliberin- A.Group C: similar to Grop B + daily injection of progesterone after that blood samples was collected and centrifuged.plasma were analysed by ELISA.the analysis of this study uses SPSS data software package and compared between the mean and LS Means LSD test at 5% significance level was used.The results of this study shows that maximum of progesterone plasma levels were in the control gruop (P ≥ 0.05).Therefore, daily injection of progesterone inhibit the growth CL. the most estrogen levels in plasma were in Group C (P ≥ 0.05) thus it can be concluded, rise in endogenous estrogen concentrations normally stimulates the preovulatory LH release in heifers.

Keywords: Luliberin- A, Cloprostenol Sodium, estrogen, progesterone, dairy heifers

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Authors: Joel Ballesteros, Harriet Jane Caleja, Florian Del Mundo, Cherrie Pascual

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Diffraction-based sensing was utilized in the quantification of human ferritin in blood serum to provide an alternative to label-based immunoassays currently used in clinical diagnostics and researches. The diffraction intensity was measured by the diffractive optics technology or dotLab™ system. Two methods were evaluated in this study: direct immunoassay and direct sandwich immunoassay. In the direct immunoassay, human ferritin was captured by human ferritin antibodies immobilized on an avidin-coated sensor while the direct sandwich immunoassay had an additional step for the binding of a detector human ferritin antibody on the analyte complex. Both methods were repeatable with coefficient of variation values below 15%. The direct sandwich immunoassay had a linear response from 10 to 500 ng/mL which is wider than the 100-500 ng/mL of the direct immunoassay. The direct sandwich immunoassay also has a higher calibration sensitivity with value 0.002 Diffractive Intensity (ng mL-1)-1) compared to the 0.004 Diffractive Intensity (ng mL-1)-1 of the direct immunoassay. The limit of detection and limit of quantification values of the direct immunoassay were found to be 29 ng/mL and 98 ng/mL, respectively, while the direct sandwich immunoassay has a limit of detection (LOD) of 2.5 ng/mL and a limit of quantification (LOQ) of 8.2 ng/mL. In terms of accuracy, the direct immunoassay had a percent recovery of 88.8-93.0% in PBS while the direct sandwich immunoassay had 94.1 to 97.2%. Based on the results, the direct sandwich immunoassay is a better diffraction-based immunoassay in terms of accuracy, LOD, LOQ, linear range, and sensitivity. The direct sandwich immunoassay was utilized in the determination of human ferritin in blood serum and the results are validated by Chemiluminescent Magnetic Immunoassay (CMIA). The calculated Pearson correlation coefficient was 0.995 and the p-values of the paired-sample t-test were less than 0.5 which show that the results of the direct sandwich immunoassay was comparable to that of CMIA and could be utilized as an alternative analytical method.

Keywords: biosensor, diffraction, ferritin, immunoassay

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Authors: Varsha Salian, Shama Rao, N. Narendra, B. Mohana Kumar

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Evolving evidence proposes the existence of a highly tumorigenic subpopulation of undifferentiated, self-renewing cancer stem cells, responsible for exhibiting resistance to conventional anti-cancer therapy, recurrence, metastasis and heterogeneous tumor formation. Importantly, the mechanisms exploited by cancer stem cells to resist chemotherapy are very less understood. Oral squamous cell carcinoma (OSCC) is one of the most regularly diagnosed cancer types in India and is associated commonly with alcohol and tobacco use. Therefore, the isolation and in vitro characterization of cancer stem-like cells from patients with OSCC is a critical step to advance the understanding of the chemoresistance processes and for designing therapeutic strategies. With this, the present study aimed to establish and characterize cancer stem-like cells in vitro from OSCC. The primary cultures of cancer stem-like cell lines were established from the tissue biopsies of patients with clinical evidence of an ulceroproliferative lesion and histopathological confirmation of OSCC. The viability of cells assessed by trypan blue exclusion assay showed more than 95% at passage 1 (P1), P2 and P3. Replication rate was performed by plating cells in 12-well plate and counting them at various time points of culture. Cells had a more marked proliferative activity and the average doubling time was less than 20 hrs. After being cultured for 10 to 14 days, cancer stem-like cells gradually aggregated and formed sphere-like bodies. More spheroid bodies were observed when cultured in DMEM/F-12 under low serum conditions. Interestingly, cells with higher proliferative activity had a tendency to form more sphere-like bodies. Expression of specific markers, including membrane proteins or cell enzymes, such as CD24, CD29, CD44, CD133, and aldehyde dehydrogenase 1 (ALDH1) is being explored for further characterization of cancer stem-like cells. To summarize the findings, the establishment of OSCC derived cancer stem-like cells may provide scope for better understanding the cause for recurrence and metastasis in oral epithelial malignancies. Particularly, identification and characterization studies on cancer stem-like cells in Indian population seem to be lacking thus provoking the need for such studies in a population where alcohol consumption and tobacco chewing are major risk habits.

Keywords: cancer stem-like cells, characterization, in vitro, oral squamous cell carcinoma

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Authors: Samad Lotfollahzadeh

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To evaluate the effect of intramuscular administration of vitamin A during dry period in pregnant dairy cows, which already received it in their daily ration, on vitamin A status of neonatal calves, a total of 30 cows were randomly selected and divided to two main groups; treatment and control group. Animals in the treatment group were subdivided into two groups. Single intramuscular injection of 2000000 IU vitamin A; was carried in 10 dairy cows at 7 months of pregnancy (group 1). In the second group of treated animals (10 cows) the injection was performed in 8 months of pregnancy (group 2). Ten pregnant dairy cows were received saline injection as placebo and selected as the control group. Blood samples were collected from experimental dairy cows at 7 and 8 months of pregnancy as well as their newborn calves’ pre and after colostrum intake. There was no significant difference between vitamin A and β- carotene concentration of dairy cows of three groups in two last months of pregnancy (P > 0.05). Vitamin A concentration of calves of two treatment groups before and after receiving of colostrum were significantly higher than that in the control group (P < 0.05). There was no significant difference between serum concentrations of vitamin A in calves of two treated groups (P > 0.05). β- Carotene concentration of serum samples of dairy cows and neonatal calves of three groups were not significantly different as compared with together. From results of the present study it can be concluded that daily supplementation of vitamin A in late pregnancy in dairy cows may not compensate the calves need for vitamin A and single injection of this vitamin A during dry either in 7 or 8 months of pregnancy can significantly increase level of vitamin A in their colostrum and neonatal calves.

Keywords: dry cow, beta carotene, newborn calves, vitamin A, dry cows

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Authors: Qiaofeng Guo

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Aims: Obesity is often accompanied by insulin resistance (IR) and whole-body inflammation. Aerobic exercise is an effective treatment to improve insulin resistance and inflammation. However, the anti-inflammatory mechanisms of exercise on epididymal and subcutaneous adipose remain to be elucidated. Here, we compared the macrophage polarization between epididymal and subcutaneous adipose after aerobic exercise. Methods: Male C57BL/6 mice were fed a normal diet group or a high-fat diet group for 12 weeks and performed aerobic training on a treadmill at 55%~65% VO₂ max for eight weeks. Food intake, body weight, and fasting blood glucose levels were monitored weekly. The intraperitoneal glucose tolerance test was to evaluate the insulin resistance model. Fat mass, blood lipid profile, serum IL-1β, TNF-α levels, and CD31/CD206 rates were analysed after the intervention. Results: FBG (P<0.01), AUCIPGTT (P<0.01), and HOMA-IR (P<0.01) increased significantly for a high-fat diet and decreased significantly after the exercise. Eight weeks of aerobic exercise attenuated HFD-induced weight gain and glucose intolerance and improved insulin sensitivity. Serum IL-1β, TNF-α, CD11C/CD206 expression in subcutaneous adipose tissue were not changed before and after exercise, but not in epididymal adipose tissue (P<0.01). Conclusion: Insulin resistance is not accompanied by chronic inflammation and M1 polarization of subcutaneous adipose tissue macrophages in high-fat diet mice. Aerobic exercise effectively improved lipid metabolism and insulin sensitivity, which may be closely associated with reduced M1 polarization of epididymal adipose macrophages.

Keywords: aerobic exercise, insulin resistance, chronic inflammation, adipose, macrophage polarization

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Authors: Shokouh Momeni, Mohammad Reza Salamat, Ali Asghar Rastegari

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Background: Osteoporosis is the most prevalent metabolic bone disease in postmenopausal women associated with reduced bone mass and increased bone fracture. Measuring bone density in the lumbar spine and hip is a reliable measure of bone mass and can therefore specify the risk of fracture. Dual-energy X-ray absorptiometry(DXA) is an accurate non-invasive system measuring the bone density, with low margin of error and no complications. The present study aimed to investigate the relationship between biochemical parameters with bone density in postmenopausal women. Materials and methods: This cross-sectional study was conducted on 87 postmenopausal women referred to osteoporosis centers in Isfahan. Bone density was measured in the spine and hip area using DXA system. Serum levels of calcium, phosphorus, alkaline phosphatase and magnesium were measured by autoanalyzer and serum levels of vitamin D were measured by high-performance liquid chromatography(HPLC). Results: The mean parameters of calcium, phosphorus, alkaline phosphatase, vitamin D and magnesium did not show a significant difference between the two groups(P-value>0.05). In the control group, the relationship between alkaline phosphatase and BMC and BA in the spine was significant with a correlation coefficient of -0.402 and 0.258, respectively(P-value<0.05) and BMD and T-score in the femoral neck area showed a direct and significant relationship with phosphorus(Correlation=0.368; P-value=0.038). There was a significant relationship between the Z-score with calcium(Correlation=0.358; P-value=0.044). Conclusion: There was no significant relationship between the values ​​of calcium, phosphorus, alkaline phosphatase, vitamin D and magnesium parameters and bone density (spine and hip) in postmenopaus

Keywords: osteoporosis, menopause, bone mineral density, vitamin d, calcium, magnesium, alkaline phosphatase, phosphorus

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Authors: Ahmet Uyar, Zabit Yener, Abdulahad Dogan

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(1). The ameliorative potential and antioxidant capacity of an extract of Urtica dioica seeds (UDS) were investigated using histopathological changes in liver and kidney of broiler, measuring serum marker enzymes, antioxidant defence systems and lipid peroxidation (malondialdehyde (MDA)) content in various tissues of broilers exposed to aflatoxin (AF). (2). A total of 32 broilers were divided randomly into 4 groups: control, UDS extract-treated, AF-treated and AF+UDS extract-treated. Broilers in control and UDS extract-treated groups were fed on a diet without AF. The AF-treated group and AF+UDS extract-treated groups were treated with an estimated 1 mg total AF/kg feed. The AF+UDS extract groups received in addition 30 ml UDS extract/kg diet for 21 days. (3). The AF-treated group had significantly decreased body weight gain when compared to the other groups. (4). Biochemical analysis showed a small increase in the concentrations of serum aspartate aminotransferase, alanine aminotransferase, gamma glutamyl transpeptidase and lactate dehydrogenase in the AF-treated group compared to that of the control group, whereas concentrations of these enzymes were decreased in the AF+UDS group compared to that of the AF-treated group. (5). Administration of supplementary UDS extract helped restore the AF-induced increase in MDA and reduced the antioxidant system towards normality, particularly in the liver, brain, kidney and heart. Hepatorenal protection by UDS extracts was further supported by the almost normal histology in AF +UDS extract-treated group as compared to the degenerative changes in the AF-treated broilers. (6). It was concluded that UDS extract has a protective hepatorenal effect in broilers affected by aflatoxicosis, probably acting by promoting the antioxidative defence systems.

Keywords: aflatoxicosis, biochemistry, broiler, histopathology, Urtica dioica seed extract

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Authors: Suruchi Shukla, Shantanu Prakash, Amita Jain

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Introduction: Arboviral diseases are one of the most common causes of viral hemorrhagic fever (VHF). Of which, Dengue and Chikungunya pose a significant health problem in India. Arbovirus has a tendency to cross the territories and emerge in the new region. Considering the above issues, in the current study active surveillance was conducted among viral hemorrhagic fever (VHF) cases reported from Uttar Pradesh (UP), India. We studied the arboviral etiology of VHF; mainly Dengue, Chikungunya, and ZIKA. Methods: Clinical samples of 465 suspected VHF cases referred to tertiary care referral center of UP, India were enrolled in the study during a period from 15th May 2016 to 9th March 2018. Serum specimens were collected and analyzed for the presence of Dengue, Chikungunya, and ZIKA either by serology and/or by molecular assays. Results: Of all tested, 165 (35.4%) cases were positive for either Dengue or Chikungunya. Dengue (21.2%) was found to be the most prevalent, followed by Chikungunya, (6.6%). None of the cases tested positive for ZIKA virus. Serum samples of 35 (7.5%) cases were positive for both Dengue and Chikungunya. DEN-2 serotype was the most predominant serotype. Phylogenetic and sequence analysis of DEN-2 strains showed 100% clustering with the Cosmopolitan genotype strain. Bleeding from several sites, jaundice, abdominal pain, arthralgia, haemoconcentration, and thrombocytopenia were significantly higher in dengue hemorrhagic cases. However, the rash was significantly more common in Chikungunya patients. Most of the Dengue and Chikungunya positive cases (Age group 6-40 years) were seen in post monsoon season (September to November). Conclusion: Only one-third of total VHF cases are positive for either Dengue/Chikungunya or both. This necessitates the screening of other etiologies capable of causing hemorrhagic manifestations.

Keywords: viral hemorrhagic fever, dengue, chikungunya, zika, India

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Authors: Amita Jain, Shantanu Prakash, Suruchi Shukla

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Introduction: Acute encephalitis syndrome (AES) is a complex multi etiology syndrome posing a great public health problem in the northern part of India. Japanese encephalitis (JE) virus is an established etiology of AES in this region. Recently, Scrub typhus (ST) is being recognized as an emerging aetiology of AES in JE endemic belt. This study was conducted to establish the direct evidence of Central nervous system invasion by Orientia tsutsugamushi leading to AES. Methodology: A total of 849 cases with clinical diagnosis of AES were enrolled from six districts (Deoria and its adjoining area) of the traditional north Indian Japanese encephalitis (JE) belt. Serum and Cerebrospinal fluid samples were collected and tested for major agent causing acute encephalitis. AES cases either positive for anti-ST IgM antibodies or negative for all tested etiologies were investigated for ST-DNA by real-time PCR. Results: Of these 505 cases, 250 patients were laboratory confirmed for O. tsutsugamushi infection either by anti-ST IgM antibodies positivity (n=206) on serum sample or by ST-DNA detection by real-time PCR assay on CSF sample (n=2) or by both (n=42).Total 29 isolate could be sequenced for 56KDa gene. Conclusion: All the strains were found to cluster with Gilliam strains. The majority of the isolates showed a 97–99% sequence similarity with Thailand and Cambodian strains. Gilliam strain of O.tsusugamushi is an emerging as one of the major aetiologies leading to AES in northern part of India.

Keywords: acute encephalitis syndrome, O. tsutsugamushi, Gilliam strain, North India, cerebrospinal fluid

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Authors: Ahmed A. M. Shoaib, Heba A. Esaily, Mahmoud M. Emara, Eman A. E. Badr, Amany S. Khalifa, Mayada M. M., Abdel-Raizk

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Background: DM is associated with metabolic bone diseases, osteoporosis, low-impact fractures and falls in geriatrics. Fetuin-A, which is a serum protein produced by the liver and promotes bone mineralization, is an independent risk factor for type 2 diabetes. Aim: Evaluation of fetuin-A level and bone mineral density in postmenopausal Egyptian female patients with type 2 diabetes mellitus and their correlation with each other & with other metabolic parameters. Patients and methods: Seventy postmenopausal female patients with type II diabetes and thirty postmenopausal female as control were included in this study. Measurement of Fetuin-A together with metabolic parameters and DXA in wrist, hip and spine, ALP, CBC, FBS, PP2H and HBA1c was done in all participants. Results: - Fetuin-A level was found to be highly significant (p< 0.001) between diabetic and nondiabetic groups and negatively correlated with BMD in spine. No difference in BMD was found between patients and control groups while significant negative correlation was found between FBS and hip BMD (<0.05) and between 2hpp and HBA1c with spine BMD in the diabetic group (<0.05). Osteoporosis represented 12.9% in spine area and 7.2% in hip and wrist areas in diabetic patients, while osteopenia were found in 58.5%, 57.1%, and 37.1% in diabetic patients in spine, wrist, and hip respectively. Conclusion: - type II diabetes cannot be considered as a risk factor for osteoporosis; while glycemic parameters (FBS, 2hpp & HBA1c) and serum Fetuin-A levels were correlated with BMD in diabetics. Good glycemic control can be protective against osteoporosis in diabetic elderly.

Keywords: fetuin-A, BMD, postmenopausal, DM type II

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