Search results for: metabolic homeostasis

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

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

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

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Authors: L. Krpalkova, N. O' Mahony, A. Carvalho, S. Campbell, S. Harapanahalli, J. Walsh

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The objective of this study was to determine the effects of environmental stressors on the performance of lactating dairy cows and discuss some future trends. There exists a relationship between the meteorological data and milk yield prediction accuracy in pasture-based dairy systems. New precision technologies are available and are being developed to improve the sustainability of the dairy industry. Some of these technologies focus on welfare of individual animals on dairy farms. These technologies allow the automatic identification of animal behaviour and health events, greatly increasing overall herd health and yield while reducing animal health inspection demands and long-term animal healthcare costs. The data set consisted of records from 489 dairy cows at two dairy farms and temperature measured from the nearest meteorological weather station in 2018. The effects of temperature on milk production and behaviour of animals were analyzed. The statistical results indicate different effects of temperature on milk yield and behaviour. The “comfort zone” for animals is in the range 10 °C to 20 °C. Dairy cows out of this zone had to decrease or increase their metabolic heat production, and it affected their milk production and behaviour.

Keywords: behavior, milk yield, temperature, precision technologies

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Authors: Patsarawadee Paojinda, Waranya Imprasittichai, Sudaratana R. Krungkrai, Nirianne Marie Q. Palacpac, Toshihiro Horii, Jerapan Krungkrai

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Fusion of the last two enzymes in the pyrimidine biosynthetic pathway in the inversed order by having COOH-terminal orotate phosphoribosyltransferase (OPRT) and NH2-terminal orotidine 5'-monophosphate decarboxylase (OMPDC), as OMPDC-OPRT, are described in many organisms. Here, we produced gene fusions of Plasmodium falciparum OMPDC-OPRT and expressed the bifunctional protein in Escherichia coli. The enzyme was purified to homogeneity using affinity and anion-exchange chromatography, exhibited enzymatic activities and functioned as a dimer. The activities, although unstable, can be stabilized by its substrate and product during purification and long-term storage. Furthermore, the enzyme expressed a perfect catalytic efficiency (kcat/Km). The kcat was selectively enhanced up to 3 orders of magnitude, while the Km was not much affected and remained at low µM levels when compared to the monofunctional enzymes. The fusion of the two enzymes, creating a “super-enzyme” with perfect catalytic power and more flexibility, reflects cryptic relationship of enzymatic reactivaties and metabolic functions on molecular evolution.

Keywords: bifunctional enzyme, orotate phosphoribosyltransferase, orotidine 5'-monophosphate decarboxylase, plasmodium falciparum

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Authors: Wizna, Helmi Muis, Hafil Abbas

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An experiment was conducted to improve the nutrient value of sago pith (Metroxylon sago Rottb) supplemented with Zn, Sulfur and urea through fermentation by using cellulolytic bacteria (Bacillus amyloliquefaciens) as inoculums. The experiment was determination of the optimum dose combination (dosage of Zn, S and urea) for sago pith fermentation based on nutrient quality and quantity of these fermented products. The study was conducted in experimental method, using the completely randomized design in factorial with 3 treatments consist of: factor A (Dose of urea: A1 = 2.0%, A2 = 3.0%), factor B (Dose of S: B1 = 0.2%, B2 = 0.4%) and factor C (Dose of Zn: C1 = 0.0025%, C2 = 0.005%). Results of study showed that optimum condition for fermentation process of sago pith with B. amyloliquefaciens caused a change of nutrient content was obtained at urea (3%), S (0,2%), and Zn (0,0025%). This fermentation process was able to increase amino acid average, reduce crude fiber content by 67% and increase crude protein by 433%, which made the nutritional value of the product based on dry matter was 18.22% crude protein, 12.42% crude fiber, 2525 Kcal/kg metabolic energy and 65.73% nitrogen retention.

Keywords: fermentation, sago pith, sulfur, Zn, urea, Bacillus amyloliquefaciens

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Authors: Marek Halenár, Eva Tvrdá, Simona Baldovská, Ľubomír Ondruška, Peter Massányi, Adriana Kolesárová

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This study aimed to assess the in vitro effects of different concentrations of the Viscum album extract on the motility, viability, and reactive oxygen species (ROS) production by rabbit spermatozoa during different time periods (0, 2, and 8h). Spermatozoa motility was assessed by using the CASA (Computer aided sperm analysis) system. Cell viability was evaluated by using the metabolic activity MTT assay, and the luminol-based luminometry was applied to quantify the ROS formation. The CASA analysis revealed that low Viscum concentrations were able to prevent a rapid decline of spermatozoa motility, especially in the case of concentrations ranging between 1 and 5 µg/mL (P<0.05 with respect to time 8h). At the same time, concentrations ranging between 1 and 100 µg/mL of the extract led to a significant preservation of the cell viability (P<0.05 in case of 5, 50 and 100 µg/mL; P<0.01 with respect to 1 and 10 µg/mL, time 8h). 1 and 5 µg/mL of the extract exhibited antioxidant characteristics, translated into a significant reduction of the ROS production, particularly notable at time 8h (P<0.01). The results indicate that the Viscum extract is capable of delaying the damage inflicted to the spermatozoon by the in vitro environment.

Keywords: CASA, mistletoe, mitochondrial activity, motility, reactive oxygen species, rabbits, spermatozoa, Viscum album

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Authors: Eva Tvrdá, Hana Greifová, Peter Ivanič, Norbert Lukáč

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The aim of this study was to evaluate the dose- and time-dependent in vitro effects of berberine (BER), a natural alkaloid with numerous biological properties on bovine spermatozoa during three time periods (0 h, 2 h, 24 h). Bovine semen samples were diluted and cultivated in physiological saline solution containing 0.5% DMSO together with 200, 100, 50, 10, 5, and 1 μmol/L BER. Spermatozoa motility was assessed using the computer assisted semen analyzer. The viability of spermatozoa was assessed by the metabolic (MTT) assay, production of superoxide radicals was quantified using the nitroblue tetrazolium (NBT) test, and chemiluminescence was used to evaluate the generation of reactive oxygen species (ROS). Cell lysates were prepared and the extent of lipid peroxidation (LPO) was evaluated using the TBARS assay. The results of the movement activity showed a significant increase in the motility during long term cultivation in case of concentrations ranging between 1 and 10 μmol/L BER (P < 0.01; P < 0.001; 24 h). At the same time, supplementation of 1, 5 and 10 μmol/L BER led to a significant preservation of the cell viability (P < 0.001; 24 h). BER addition at a range of 1-50 μmol/L also provided a significantly higher protection against superoxide (P < 0.05) and ROS (P < 0.001; P < 0.01) overgeneration as well as LPO (P < 0.01; P<0.05) after a 24 h cultivation. We may suggest that supplementation of BER to bovine spermatozoa, particularly at concentrations ranging between 1 and 50 μmol/L, may offer protection to the motility, viability and oxidative status of the spermatozoa, particularly notable at 24 h.

Keywords: berberine, bulls, motility, oxidative profile, spermatozoa, viability

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Authors: Mrinal Gupta, Mohammad Rumman, Babita Singh Abbas Ali Mahdi, Shivani Pandey

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Introduction: Berberine (BBR), a bioactive compound isolated from Coptidis Rhizoma, possesses diverse pharmacological activities, including anti-bacterial, anti-inflammatory, antitumor, hypolipidemic, and anti-diabetic. However, its role as an anti-diabetic agent in animal models of dexamethasone (Dex)-induced diabetes remains unknown. Studies have shown that natural compounds, including aloe, caper, cinnamon, cocoa, green and black tea, and turmeric, can be used for treating Type 2 diabetes mellitus (DM). Compared to conventional drugs, natural compounds have fewer side effects and are easily available. Herein, we studied the anti-diabetic effects of BBR in a mice model of Dex-induced diabetes. Methods: HepG2 cell line was used for glucose release and glycogen synthesis studies. Cell proliferation was measured by methylthiotetrazole (MTT) assay. For animal studies, mice were treated with Dex (2 mg/kg, i.m.) for 30 days and the effect of BBR at the doses 100, 200, and 500 mg/kg (p.o.) was analyzed. Glucose, insulin, and pyruvate tests were performed to evaluate the development of the diabetic model. An echo MRI was performed to assess the fat mass. Further, to elucidate the mechanism of action of BBR, mRNA expression of genes regulating gluconeogenesis, glucose uptake, and glycolysis were analyzed. Results: In vitro BBR had no impact on cell viability up to a concentration of 50μM. Moreover, BBR suppressed the hepatic glucose release and improved glucose tolerance in HepG2 cells. In vivo, BBR improved glucose homeostasis in diabetic mice, as evidenced by enhanced glucose clearance, increased glycolysis, elevated glucose uptake, and decreased gluconeogenesis. Further, Dex treatment increased the total fat mass in mice, which was ameliorated by BBR treatment. Conclusion: BBR improves glucose tolerance by increasing glucose clearance, inhibiting hepatic glucose release, and decreasing obesity. Thus, BBR may become a potential therapeutic agent for treating glucocorticoid-induced diabetes and obesity in the future.

Keywords: glucocorticoid, hyperglycemia, berberine, HepG2 cells, insulin resistance, glucose

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Authors: Hai Chi, Ibrahim Mehmeti, Kirill Ovchinnikov, Hegle Holo, Ingolf F. Nes, Dzung B. Diep

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By using a panel of multiple indicator strains of different bacterial species and genera, we screened a large collection of bacterial isolates (over 1800 isolates) derived from raw milk, for bacteriocin producers with broad inhibition spectra (BIS). Fourteen isolates with BIS were identified, and by 16S rDNA sequencing they were found to belong to Lactococcus garvieae (10 isolates) and Enterococcus feacalis (4 isolates). Further analysis of the ten L. garvieae isolates revealed that they were very similar, if not identical, to each other in metabolic and genetic terms: they had the same fermentation profile on different types of sugars, repetitive sequence-based PCR (rep-PCR) DNA pattern as well as they all had the same inhibition profile towards over 50 isolates of different species. The bacteriocin activity from one of the L. garvieae isolates was assessed further. The bacteriocin which was termed garvicin KS, was found to be heatstable and proteinase-labile and its inhibition spectrum contained many distantly related genera of Firmicutes, comprising most lactic acid bacteria (LAB) as well as problematic species of Bacillus, Listeria, Streptococcus and Staphylococcus and their antibiotic resistant derivatives (e.g. VRE, MRSA). Taken together, the results indicate that this is a potent bacteriocin from L. garvieae and that its very broad inhibition spectrum can be a very useful property for use in food preservation as well as in infection treatments caused by gram-positive pathogens and their antibiotic-derivatives.

Keywords: bacteriocin, lactic acid bacteria, Lactococcus garvieae, antibiotics resistance

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Authors: Urarat Nanna, Jarinyaporn Naowaboot

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Tiliacora triandra (T. triandra) is traditional Southeast Asian medicine and widely used in the cuisines of northeast Thailand and Laos. It has been used as antipyretic, detoxication agent, antiinflammation. But the activity of T. triandra leaf water extract (TTW) in the regulation of metabolic syndrome is still little known. In this study, we evaluated the effects of TTW in high-fat diet (HFD)-induced obese mice. Male ICR mice were induced to be obese by HFD feeding (45 kcal% lard fat) for 12 weeks. During the last 6 weeks of diet feeding, the obese mice were treated with TTW at 250 and 500 mg/kg/day. The biochemical parameters and histology analysis were measured at the end of treatment period. After 6 weeks of TTW treatment, the hyperglycemia, hyperinsulinemia, hyperleptinemia and hyperlipidemia were significantly decreased. Hepatic lipid accumulation and adipocyte hypertrophy were also reduced. Serum adiponectin was increased in TTW-treated obese mice. TTW treatment could reduce the malondialdehyde in serum and liver tissue. Furthermore, the elevated serum inflammatory cytokines, tumor necrosis factor-α (TNF-α) and monocyte chemoattractant protein-1 were reduced (MCP-1) by TTW. These results suggest that T. triandra leaf is a beneficial plant in alleviating hyperglycemia, hyperlipidemia, oxidative stress and inflammation in the obese condition induced by HFD.

Keywords: Tiliacora triandra, insulin resistance, hyperlipidemia, oxidative stress

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Authors: Mohammad Najmuddin Khan, Mohamad Khaleel Albalwi

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The prevalence of obesity has risen dramatically in past several decades. Hormonal and genetic factors are rarely the cause of childhood obesity. Because obese adult may suffer life-long physical and emotional consequences, it is imperative to discuss prevention with parents during well-child examinations. Purpose of the study was to compare the serum lipid profile of obese and non-obese males. Twenty two male students were selected from Al-Jouf University. Their age ranged from 19 to 29. They were divided into groups. One group (N=15) having more than 20% fat was considered as obese group, another group (N=7) was considered as non-obese group. Fasting blood samples were analysed for blood cholesterol, triglycerides, low density lipoprotein cholesterol (LDL-C) and high density lipoprotein cholesterol (HDL-C). Independent test was applied to compare mean difference. In obese group, significantly higher cholesterol and triglycerides were observed. On the contrary, obese group had significantly lower HDL-C concentration than the non-obese group. The adult obese has relatively larger changes in serum lipids at any given level of obesity. On the average, higher amount of fat makes it more likely for an individual to be dyslipidemic and to express elements of the metabolic syndrome. Increased triglycerides level in obese impaired lipolysis which reduced the HDL-C concentrations.

Keywords: obesity, serum lipid profile, Al-Jouf, HDL, LDL

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Authors: Dina A. Selim, Eman Shawky, Rasha M. Abu El-Khair

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In the current study, UPLC-MS/MS combined to chemometrics were applied on seven Sri Lankan tea grades; Orange Pekoe, Flowery Pekoe, Broken Orange Pekoe Fannings, Broken Orange Pekoe black tea, green tea, silver tips and golden tips white tea grades for their comprehensive metabolic profiling. Certain metabolites, namely, Theasensinin C and E, theaflavin and theacitrin appeared to be the main chemical markers of black tea type, catechin, epicatechin, epigallocatechin, methyl epigallocatechin were the main discriminatory markers of green tea type, while theanine, oolongotheanine and quercetin glycosides were the main chemical markers of white tea type. Theogalloflavin, epigallocatechin and flavonoid glycosides were the main down-accumulated metabolites while theaflavin gallate, and N-ethyl pyrrolidinone epicatechin were the chief up- accumulated metabolites between whole and broken black tea leave grades while puerin A and C and gallic acid was the main down- accumulated metabolites and N-ethyl pyrrolidinone epicatechin gallate was the main up-accumulated one between broken and fanning black tea grades.

Keywords: tea grading, Sri Lankan tea, chemometrics, metabolomics, chemical markers

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Authors: V. K. Srivastava

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The chemistry of compounds containing transition metals bound to sulfur containing ligands has been actively studied. Interest in these compounds arises from the identification of the biological importance of iron-sulfur containing proteins as well as the unusual behaviour of several types of synthetic metal-sulfur complexes. Metal complexes (C₆H₅)₄P)₂ Pt(Mos₄)₂, (C₆H₅)₄P)₂ Pd(MoS₄)₂, (C₆H₅)₄P)₂ Ni(MoS₄)₂ of bioinorganic relevance were investigated. The complexes [M(M'S₄)₂]²⁻ were prepared with high yield and purity as salts of the variety of organic cations. The diamagnetism and spectroscopic properties of these complexes confirmed that their structures are essentially equivalent with two bidentate M'S₄²⁻ ligands coordinated to the central d⁸ metal in a square planer geometry. The interaction of the complexes with CT-DNA was studied. Results showed that metal complexes increased DNA's relative viscosity and quench the fluorescence intensity of EB bound to DNA. In antimicrobial activities, all complexes showed good antimicrobial activity higher than ligand against gram positive, gram negative bacteria and fungi. The antitumor properties have been tested in vitro against two tumor human cell lines, Hela (derived from cervical cancer) and MCF-7 (derived from breast cancer) using metabolic activity tests. Result showed that the complexes are promising chemotherapeutic alternatives in the search of anticancer agents.

Keywords: anti cancer, biocidal, DNA binding, spectra

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Authors: Adel Omar Ashour, Paul S. Dyer

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Aspergillus nidulans can reproduce by asexual or sexual means, producing green conidiospores or red-purple ascospores respectively. The latter one is produced in dark-purple globose ‘cleistothecia’ which are surrounded by Hülle cells. The species has a homothallic (self fertile) sexual breeding system. Given the extra metabolic costs associated with sexual compared to asexual reproduction it would be predicted that ascospore production would confer evolutionary benefits. However, due to the homothallic breeding system there is very rarely any increased genetic variation in ascospore offspring and traditionally conidia and ascospores are considered to be equally environmental resistant. We therefore examined in detail whether conidia and ascospores might exhibit as yet undetected differences in spore viability when subjected to certain environmental stressors. Spores from two strains of A. nidulans (comprising wild-type and KU mutants) were exposed to various levels of temperature (50-70°C for 30 min) and UV (350 nm for 10-60 min) stress. Results of experiments will be presented, including comparison of ‘D’ (decimal point reduction) values of conidia versus ascospores of A. nidulans. We detected that under certain exposure levels ascospores have significantly increased resistance compared to conidia. The increased environmental resistance of ascospores might be a key factor explaining the persistence of sexuality in this homothallic species, and reasons for differential survival are suggested.

Keywords: Aspergillus nidulans, asexual reproduction, conidia, ascospores, cleistothecia, d-value

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Authors: Tahmineh Hasanzadeh, Afshin Farahbakhsh

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There is some report about the hypoglycemic effect of Juglans rejia L. leaf in alloxan induced diabetic rats and hypoglycemic effect of its fruit peel administered intraperitoneally.In Iranian traditional medicine, septum of walnut shell (SWS) was recommended to reduce blood glucose. For this purpose, 41 male bulb/C mice 25-30 gm were divided into five groups. All the animals received IP injection of streptozotocin (STZ) (220 mg/kg). Two weeks later, the diabetic animals were received daily oral treatment of normal saline and aqueous extract of SWS (200, 400, 600 and 800 mg/kg) respectively for four weeks. Blood samples were taken from retro orbital sinus before the start of the experiment and repeated each two weeks. At the end of the experiment, the animals were sacrificed and the pancreatic tissues were fixed, prepared and stained by Hematoxylin-Eosin for light microscope studies. The results showed that in each group, the SWS extract reduced blood glucose in a long time (p < 0.05). metabolic extract in STZ- induced diabetic rats, which was accompanied by the hypoglycemic effect of leaf extract. However, this effect should be determined with scientific researches. Therefore, the aim of this study is to evaluate the effect of the aqueous extract of SWS on blood glucose and histopathological structure of pancreas.

Keywords: septum of walnut, blood glucose, pancreas, diabetes, walnut leaf, walnut peel, insulin

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Authors: Daria Reczynska, Justyna Jarczak, Michal Czopowicz, Danuta Sloniewska, Karina Horbanczuk, Wieslaw Jarmuz, Jaroslaw Kaba, Emilia Bagnicka

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Since people, animals and plants are constantly exposed to pathogens they have developed very complex systems of defense. Among ca. 1000 antimicrobial peptides from different families so far identified, approximately 30 belonging to cathelicidin family can be found in mammals. Cathelicidins probably constitute the first line of defense because they can act at a physiological salt concentration which is present in healthy tissues. Moreover, the low salt concentration which is present in infected tissues inhibits their activity. In goat bactenecin 7.5 (BAC7.5), bactenecin 5 (BAC5), myeloid antimicrobial peptide 28 (MAP28), myeloid antimicrobial peptide 34 (MAP34 A and B), goat bactenecin3.4 (ChBac3.4) were identified. Caprine arthritis-encephalitis (CAE) caused by small ruminant lentivirus (SRLV) is economic problem. The main CAE symptoms are weight loss, arthritis, pneumonia and mastitis (significant elevation of the somatic cell count and deterioration of some technological parameters). The study was conducted on 24 dairy goats. The animals were divided into two groups: experimental (SRLV-infected) and control (non-infected). The blood samples were collected five times: on the 1st, 7th, 30th, 90th and 150thday of lactation. The levels of transcripts of BAC7.5, BAC5, MAP28 and MAP34 genes in blood leucocytes were measured using qPCR method. There were no differences in mRNA levels of studied genes between stages of lactation. The differences were observed in expressions of BAC5, MAP28 and MAP34 genes with lower levels in the experimental group. There was no difference in BAC7.5 expression between groups. The decreased levels of transcripts of cathelicidin genes in blood leucocytes of SRLV-infected goats may indicate the disturbances of homeostasis in organisms. It can be concluded that SRLV infection seems to inhibit expression of cathelicidin genes. The study was financed by a grant from the National Scientific Center No. UMO-2013/09/B/NZ/03514.

Keywords: goat, CAEV, cathelicidins, blood leukocytes, gene expression

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Authors: Xiaobo Wang, Zhen-Yu Chen

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Higher plasma total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-C) are independent risk factors of cardiovascular disease while high-density lipoprotein cholesterol (HDL-C) is protective. Vitamin D is well-known for its regulatory role in calcium homeostasis. Its potential important role in cardiovascular disease has recently attracted much attention. This study was conducted to investigate effects of different dosage of calcitriol on plasma lipoprotein profile and the underlying mechanism. Sixty male Syrian Golden hamsters were randomly divided into 6 groups: no-cholesterol control (NCD), high-cholesterol control (HCD), groups with calcitriol supplementation at 10/20/40/80ng/kg body weight (CA, CB, CC, CD), respectively. Calcitriol in medium-chain triacylglycerol (MCT) oil was delivered to four experimental groups via oral gavage every other day, while NCD and HCD received MCT oil in the equivalent amount. NCD hamsters were fed with non-cholesterol diet while other five groups were maintained on diet containing 0.2% cholesterol to induce a hypercholesterolemic condition. The treatment lasts for 6 weeks followed by sample collection after hamsters sacrificed. Four experimental groups experienced a reduction in average food intake around 11% compared to HCD with slight decrease in body weight (not exceeding 10%). This reduction reflects on the deceased relative weights of testis, epididymal and perirenal adipose tissue in a dose-dependent manner. Plasma calcitriol levels were measured and was corresponding to oral gavage. At the end of week 6, lipoprotein profiles were improved with calcitriol supplementation with TC, non-HDL-C and plasma triglyceride (TG) decreased in a dose-dependent manner (TC: r=0.373, p=0.009, non-HDL-C: r=0.479, p=0.001, TG: r=0.405, p=0.004). Since HDL-C of four experiment groups showed no significant difference compared to HCD, the ratio of nHDL-C to HDL-C and HDL-C to TC had been restored in a dose-dependent manner. For hamsters receiving the highest level of calcitriol (80ng/kg) showed a reduction of TC by 11.5%, nHDL-C by 24.1% and TG by 31.25%. Little difference was found among six groups on the acetylcholine-induced endothelium-dependent relaxation or contraction of thoracic aorta. To summarize, calcitriol supplementation in hamster at maximum 80ng/kg body weight for 6 weeks lead to an overall improvement in plasma lipoprotein profile with decreased TC and TG level. The molecular mechanism of its effects is under investigation.

Keywords: cholesterol, vitamin D, calcitriol, hamster

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Authors: Thouraya Majoul

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Obesity is now a real public health issue throughout the world, and it is well-established that obesity leads to cardiovascular diseases. The prevention and treatment of obesity using nutritional supplements has become a realistic and effective approach. This study was carried out to analyze the incidence of a high-fat diet on rat heart metabolism as well as on fatty acids composition, then to investigate the eventual protective effects of a grape seed extract (GSE). The experimental design consisted of three rat groups subjected to three different conditions; standard (SD), high-fat diet (HFD) and HFD+GSE (HG). We showed that GSE counteracted the effect of HFD on fatty acid composition, namely, docosapentaenoic acid, docosahexaenoic acid, arachidonic acid (ARA), palmitic acid (PA) and palmitoleic acid. Besides, GSE treatment restored HFD-altered metabolic pathways through the recovery of some cardiac enzyme activities such as lipase, glucose 6 phosphate dehydrogenase and pyruvate dehydrogenase. The cardiac lactate level and lactate dehydrogenase activity were also analyzed in relation to HFD and GSE administration. To our knowledge, this is the first study showing the anti-obesity and cardioprotective effects of GSE in relation to fatty acid composition and some cardiac enzymes, supporting its role as a therapeutic agent of obesity.

Keywords: Grape seed extract, phenolic, obesity, cardioprotective, lipotoxicity, energy metabolism

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Authors: Marek Halenár, Eva Tvrdá, Tomáš Slanina, Ľubomír Ondruška, Eduard Kolesár, Peter Massányi, Adriana Kolesárová

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The present in vitro study was designed to reveal whether amygdalin (AMG) is able to cause changes to the motility, viability and mitochondrial activity of rabbit spermatozoa. New Zealand White rabbits (n = 10) aged four months were used in the study. Semen samples were collected from each animal and used for the in vitro incubation. The samples were divided into five equal parts and diluted with saline supplemented with 0, 0.5, 1, 2.5 and 5 mg/mL AMG. At times 0h, 3h and 5h spermatozoa motion parameters were assessed using the SpermVision™ computer-aided sperm analysis (CASA) system, cell viability was examined with the metabolic activity (MTT) assay, and the eosin-nigrosin staining technique was used to evaluate the viability of rabbit spermatozoa. All AMG concentrations exhibited stimulating effects on the spermatozoa activity, as shown by a significant preservation of the motility (P<0.05 with respect to 0.5 mg/mL and 1 mg/mL AMG; Time 5 h) and mitochondrial activity (P< 0.05 in case of 0.5 mg/mL AMG; P< 0.01 in case of 1 mg/mL AMG; P < 0.001 with respect to 2.5 mg/mL and 5 mg/mL AMG; Time 5 h). None of the AMG doses supplemented had any significant impact of the spermatozoa viability. In conclusion, the data revealed that short-term co-incubation of spermatozoa with AMG may result in a higher preservation of the sperm structural integrity and functional activity.

Keywords: amygdalin, CASA, mitochondrial activity, motility, rabbits, spermatozoa, viability

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Authors: Mohammadjavad Sotoudeheian, Navid Farahmandian

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Cardiac hypertrophy arises in response to persistent increases in hemodynamic loads. In comparison, diabetic cardiomyopathy is defined by an abnormal myocardial changes without other cardiac-related risk factors. Pathological cardiac hypertrophy and myocardial remodeling are hallmarks of cardiovascular diseases and are risk factors for heart failure. The transcription factor forkhead box class O (FOXOs) can protect heart tissue by hostile oxidative stress and stimulating apoptosis and autophagy. FOXO proteins, as sensitive elements and mediators in response to environmental changes, have been revealed to prevent and inverse cardiac hypertrophy. FOXOs are inhibited by insulin and are critical mediators of insulin action. Insulin deficiency and uncontrolled diabetes lead to a catabolic state. FOXO1 acts downstream of the insulin-dependent pathways, which are dysregulated in diabetes. It regulates cardiomyocyte hypertrophy downstream of IGF1R/PI3K/Akt activation, which are critical regulators of cardiac hypertrophy. The complex network of signaling pathways comprising insulin/IGF-1 signaling, AMPK, JNK, and Sirtuins regulate the development of cardiovascular dysfunction by modulating the activity of FOXOs. Insulin receptors and IGF1R act via the PI3k/Akt and the MAPK/ERK pathways. Activation of Akt in response to insulin or IGF-1 induces phosphorylation of FOXOs. Increased protein synthesis induced by activation of the IGF-I/Akt/mTOR signaling pathway leads to hypertrophy. This pathway and the myostatin/Smad pathway are potent negative muscle development regulators. In cardiac muscle, insulin receptor substrates (IRS)-1 or IRS-2 activates the Akt signaling pathway and inactivate FOXO1. Under metabolic stress, p38 MAPK promotes degradation of IRS-1 and IRS-2 in cardiac myocytes and activates FOXO1, leading to cardiomyopathy. Sirt1 and FOXO1 interaction play an essential role in starvation-induced autophagy in cardiac metabolism. Inhibition of Angiotensin-II induced cardiomyocyte hypertrophy is associated with reduced FOXO1 acetylation and activation of Sirt1. The NF-κB, ERK, and FOXOs are de-acetylated by SIRT1. De-acetylation of FOXO1 induces the expression of genes involved in autophagy and stimulates autophagy flux. Therefore, under metabolic stress, FOXO1 can cause diabetic cardiomyopathy. The overexpression of FOXO1 leads to decreased cardiomyocyte size and suppresses cardiac hypertrophy through inhibition of the calcineurin–NFAT pathway. Diabetes mellitus is associated with elevation of O-GlcNAcylation. Some of its binding partners regulate the substrate selectivity of O-GlcNAc transferase (OGT). O-GlcNAcylation of essential contractile proteins may inhibit protein-protein interactions, reduce calcium sensitivity, and modulate contractile function. Uridine diphosphate (UDP)-GlcNAc is the obligatory substrate of OGT, which catalyzes a reversible post-translational protein modification. The increase of O-GlcNAcylation is accompanied by impaired cardiac hypertrophy in diabetic hearts. Inhibition of O-GlcNAcylation blocks activation of ERK1/2 and hypertrophic growth. O-GlcNAc modification on NFAT is required for its translocation from the cytosol to the nucleus, where NFAT stimulates the transcription of various hypertrophic genes. Inhibition of O-GlcNAcylation dampens NFAT-induced cardiac hypertrophic growth. Transcriptional activity of FOXO1 is enriched by improved O-GlcNAcylation upon high glucose stimulation or OGT overexpression. In diabetic conditions, the modification of FOXO1 by O-GlcNAc is promoted in cardiac troponin I and myosin light chain 2. Therefore targeting O-GlcNAcylation represents a potential therapeutic option to prevent hypertrophy in the diabetic heart.

Keywords: diabetes, cardiac hypertrophy, O-GlcNAcylation, FOXO1, Akt, PI3K, AMPK, insulin

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Authors: Min-Ju Jo, Min-Young Um, Moonsung Choi, Sooim Shin

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Type 2 diabetes (T2D) is characterized by insulin resistance, the inability of β-cell and the dysfunction of mitochondria. To characterize effects of Cassia tora extract on mitochondrial dysfunction related T2D, the reduced glutathione level, amount of mitochondrial complexes and activities of mitochondrial complexes were measured. Three groups of mice were modeled; a control group was fed a normal diet, a diabetic group was fed a diabetic diet high in fat and carbohydrates, and a third group was fed a diabetic diet + 70% ethanol extracted Cassia tora seeds for 12 weeks. The amount of mitochondria was determined by Bradford assay after isolation of mitochondria in liver from each group. During isolation of mitochondria, cytosolic fractions of the tissue were collected to measure the reduced glutathione level. Interestingly, high level of the reduced glutathione was observed in Cassia tora treated group and decreased activities of mitochondrial complexes in Cassia tora treated group compared to the diabetic diet group. It indicates that Cassia tora has the potential to increase the reduced form of glutathione functioned as an important antioxidant in cells, and to reduce mitochondrial metabolic compensatory mechanism.

Keywords: antioxidant, Cassia tora, diabetes, electron transport chain, glutathione, mitochondria, spectrophotometry

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Authors: Berina Pilipović, Saša Pilipović, Maja Pašić-Kulenović

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Inflammation is the body's response to impaired homeostasis caused by infection, injury or trauma resulting in systemic and local effects. Inflammation causes the body's response to injury and is characterized by a series of events including inflammatory response, response to pain receptors and the recovery process. Inflammation can be acute and chronic. The inflammatory response is described in three different phases. Free radical is an atom or molecule that has the unpaired electron and is therefore generally very reactive chemical species. Biologically important example of reaction with free radicals is called Lipid peroxidation (LP). Lipid peroxidation reactions occur in biological membranes, and if at the outset is not stopped with the action of antioxidants, it will bring damage to the membrane, which results in partial or complete loss of their physiological functions. Calcium antagonists and beta-adrenergic receptor antagonists are known drugs, and for many years and widely used in the treatment of cardiovascular diseases. Some of these compounds also show antioxidant activity. The mechanism of antioxidant activities of calcium antagonists and beta-blockers is unknown, since their structure varies widely. This research investigated the possible local anti-inflammatory activity of ointments containing 1% carvedilol in the white petrolatum USP. Ear inflammation was induced by 3% croton oil acetone solution, in quantity of 10 µl on both mouse ears. Albino Swiss mouse (n = 8) are treated with 2.5 mg/ear ointment, and control group was treated on the same way as previous with hydrocortisone 1% ointment (2.5 mg/ear). The other ear of the same animal was used as control one. Ointments were administered once per day, on the left ear. After treatment, ears were observed for three days. After three days, we measured mass (mg) of 6 mm ear punch of treated and controlled ears. The results of testing anti-inflammatory effects of ointments with carvedilol in the mouse ear model show stronger observed effect than ointment with 1% hydrocortisone in the same basis. Identical results were confirmed by the difference between the mass of 6 mm ears punch. The results were also confirmed by histological examination. Ointments with carvedilol showed significant reduction of the inflammation process caused by croton oil on the mouse inflammation model.

Keywords: antioxidant, carvedilol, inflammation, mouse ear

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Authors: Chuanpeng Zhou

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The effect of dietary carbohydrate (CHO) level on serum physiological response, hepatic antioxidant abilities and heat shock protein 70 (HSP70) expression of Wuchang bream (Megalobrama amblycephala) was studied. Two isonitrogenous (28.56% crude protein) and isolipidic (5.28% crude lipid) diets were formulated to contain 30% or 53% wheat starch. Diets were fed for 90 days to fish in triplicate tanks (28 fish per tank). At the end of feeding trial, significantly higher serum triglyceride level, insulin level, cortisol level, malondialdehyde (MDA) content were observed in fish fed the 53% CHO diet, while significantly lower serum total protein content, alkaline phosphatase (AKP) activity, superoxide dismutase (SOD) activity and total antioxidative capacity (T-AOC) were found in fish fed the 53% CHO diet compared with those fed the 30% diet. The relative level of hepatic heat shock protein 70 mRNA was significantly higher in the 53% CHO group than that in the 30% CHO at 6, 12, and 48 h after feeding. The results of this study indicated that ingestion of 53% dietary CHO impacted the nonspecific immune ability and caused metabolic stress of Megalobrama amblycephala.

Keywords: Megalobrama amblycephala, carbohydrate, fasted and postprandial response, immunity, Hsp70

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Authors: Ikram Kamal, Mohamed Blaghen

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Petroleum hydrocarbons are serious problems and global pollutants in the environment due to their toxicity, carcinogenicity and persistent organic pollutant properties. One of the approaches to enhance biodegradation of petroleum hydrocarbons is to use biosurfactant. Biosurfactants are amphiphilic biomolecules produced as metabolic by-products from microorganisms they received considerable attention in the field of environmental remediation processes such as bioremediation. Biosurfactants have been considered as a desirable alternative to synthetic surfactants in various applications particularly in the environmental field. In comparison with their synthetic counterparts, biosurfactants have been reported to be less toxic, biodegradable and persistent. In this study we have investigated the potential of bacterial strains collected aseptically from the lagoon Marchika (water and soil) in Nador, Morocco; for the production of biosurfactants. This study also aimed to optimize the biosurfactant production process by changing the variables that influence the type and amount of biosurfactant produced by these microorganisms such as: carbon sources and also other physical and chemical parameters such as temperature and pH. Emulsification index, methylene blue test and thin layer chromatography (TLC) revealed the ability of strains used in this study to produce compounds that could emulsify gasoline. In addition, a HPLC/MS was used to separate and identify different biosurfactants purified.

Keywords: petroleum hydrocarbons, biosurfactants, biodegradation, lagoon marchika, emulsification index

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Authors: Abul B. M. M. K. Islam, Nuria Lopez-Bigas, Elizaveta V. Benevolenskaya

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RBP2 has shown to be important for cell differentiation control through epigenetic mechanism. The main aim of the present study is genome-wide location analysis of human RBP2 isoforms that differ in a histone-binding domain by ChIPseq. It is conceivable that the larger isoform (LI) of RBP2, which contains a specific H3K4me3 interacting domain, differs from the smaller isoform (SI) in genomic location, may account for the observed diversity in RBP2 function. To distinguish the two RBP2 isoforms, we used the fact that the SI lacks the C-terminal PHD domain and hence used the antibodies detecting both RBP2 isoforms (AI) through a common central domain, and the antibodies detecting only LI but not SI, through a C-terminal PHD domain. Overall our analysis suggests that RBP2 occupies about 77 nucleotides and binds GC rich motifs of active genes, does not bind to centromere, telomere, or enhancer regions, and binding sites are conserved compare to random. A striking difference between the only-SI and only-LI is that a large number of only-SI peaks are located in CpG islands and close to TSS compared to only-LI peaks. Enrichment analysis of the related genes indicates that several oncogenic pathways and metabolic pathways/processes are significantly enriched among only-SI/AI targets, but not LI/only-LI peak’s targets.

Keywords: bioinformatics, cancer, ChIP-seq, KDM5A

Procedia PDF Downloads 287

Authors: Sheila R. Pai, N. K. Subbalakshmi, C. Vidya

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Background: Accumulating evidence from prospective studies suggests an increased risk of type 2 diabetes associated with sleep deprivation and sleep disorders. Shift work by disrupting the circadian rhythm, could possibly cause metabolic disturbances. Objective: To investigate the influence of shift work on fasting blood glucose in hospital workers population. Materials and Methods: This was a cross-sectional study including 90 night shift workers (study group) and 90 day workers (controls) drawn from paramedical personnel. Night shift work was on a forward rotation basis, with an average of one night shift every 4 weeks. Each night shift rotation was for a period of 7 days, with a total of 8 hours of shift work per night. In the entire subjects body mass index (BMI) and fasting blood sugar (FBS) was measured. Statistical analysis included unpaired t test, Mann-Whitney ‘U’ test and Chi-square test. P value less than 0.05 was considered significant. Result: The study and control groups were comparable with regard to age, sex distribution and duration of employment. FBS was higher in study group compared to controls (p < 0.0001). There was no significant difference in BMI between control and study group. Conclusion: Shift work may adversely influence glucose metabolism.

Keywords: shift work, fasting blood sugar, sleep disturbances, diabetes

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Authors: Rim Frikha, Maha Ben Jema, Moez Elloumi, Tarek Rebai

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Background: Acute lymphoblastic leukemia (ALL); a common blood cancer characterized by the interaction between genetic and environmental factors. Methylenetetrahydrofolate reductase (MTHFR) is an essential folate metabolic enzyme in the processes of DNA synthesis and methylation. A common functional variant of the MTHFR gene, the A1298C, which induces disturbances in folate metabolism, may affect susceptibility to ALL. Objective: The present study aimed to assess the prevalence of MTHFR polymorphism A1298 > C in Tunisian children with ALL. Materials and Methods: A total of 28 Tunisian ALL children were enrolled in this study. Genomic DNA was extracted from whole venous blood collected in ethylenediaminetetraacetic acid (EDTA). Genotyping was carried out with restriction fragment length polymorphism (RFLP) using MboII restriction enzyme. Genotype distribution and allele frequency of MTHFR A1298C was calculated in ALL patients. Results: The A1298C variant of MTHFR was found in 11(19.6%) heterozygous and one homozygous patient (3.5%). Conclusions: This result highlights that A1298C polymorphism of MTHFR is common in Tunisian childhood ALL and suggests that this variant may have a potential role in leukemogenesis. Genotyping of large samples and different ethnicities are required to validate these findings.

Keywords: methylenetetrahydrofolate reductase, acute lymphoblastic leukemia, A1298C variant, prevalence

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Authors: Tayyaba Bari, Muhammad Hamza Anjum, Samra Kanwal, Fakhera Ikram

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Osteoarthritis, a degenerative condition, affects more than 213 million individuals globally. Since articular cartilage has no or limited vessels, therefore, after deteriorating, it is unable to rejuvenate. Traditional approaches for cartilage repair, like autologous chondrocyte implantation, microfracture and cartilage transplantation are often associated with postoperative complications and lead to further degradation. Decellularized human umbilical cord has gained interest as a viable treatment for cartilage repair. Decellularization removes all cellular contents as well as debris, leaving a biologically active 3D network known as extracellular matrix (ECM). This matrix is biodegradable, non-immunogenic and provides a microenvironment for homeostasis, growth and repair. UC derived bioink function as 3D scaffolding material, not only mediates cell-matrix interactions but also adherence, proliferation and propagation of cells for 3D organoids. This study comprises different physical, chemical and biological approaches to optimize the decellularization of human umbilical cord (UC) tissues followed by the solubilization of these tissues to bioink formation. The decellularization process consisted of two cycles of freeze thaw where the umbilical cord at -20˚C was thawed at room temperature followed by dissection in small sections from 0.5 to 1cm. Similarly decellularization with ionic and non-ionic detergents Sodium dodecyl sulfate (SDS) and Triton-X 100 revealed that both concentrations of SDS i.e 0.1% and 1% were effective in complete removal of cells from the small UC tissues. The results of decellularization was further confirmed by running them on 1% agarose gel. Histological analysis revealed the efficacy of decellularization, which involves paraffin embedded samples of 4μm processed for Hematoxylin-eosin-safran and 4,6-diamidino-2-phenylindole (DAPI). ECM preservation was confirmed by Alcian Blue, and Masson’s trichrome staining on consecutive sections and images were obtained. Sulfated GAG’s content were determined by 1,9-dimethyl-methylene blue (DMMB) assay, similarly collagen quantification was done by hydroxy proline assay. This 3D bioengineered scaffold will provide a typical atmosphere as in the extracellular matrix of the tissue, which would be seeded with the mesenchymal cells to generate the desired 3D ink for in vitro and in vivo cartilage regeneration applications.

Keywords: umbilical cord, 3d printing, bioink, tissue engineering, cartilage regeneration

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Authors: T. Wresdiyati, S. Sa’diah, A. Winarto

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Diabetes mellitus (DM) is a metabolic disease that can be indicated by the high level of blood glucose. The objective of this study was to observe the antidiabetic properties of ethanolic extract of Indonesian Swietenia mahagoni Jacq. seed on the profile of pancreatic superoxide dismutase and β-cells in the alloxan- experimental diabetic rats. The Swietenia mahagoni seed was obtained from Leuwiliang-Bogor, Indonesia. Extraction of Swietenia mahagoni was done by using ethanol with maceration methods. A total of 25 male Sprague dawley rats were divided into five groups; (a) negative control group, (b) positive control group (DM), (c) DM group that was treated with Swietenia mahagoni seed extract, (d) DM group that was treated with acarbose, and (e) non-DM group that was treated with Swietenia mahagoni seed extract. The DM groups were induced by alloxan (110 mg/kgBW). The extract was orally administrated to diabetic rats 500 mg/kg/BW/day for 28 days. The extract showed hypoglycemic effect, increased body weight, increased the content of superoxide dismutase in the pancreatic tissue, and delayed the rate of β-cells damage of experimental diabetic rats. These results suggested that the ethanolic extract of Indonesian Swietenia mahagoni Jacq. seed could be proposed as a potential anti-diabetic agent.

Keywords: beta cells, diabetes, hypoglycemic, rat, Swietenia mahagoni

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Authors: Monika Yadav

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The incorporation of a healthy lifestyle significantly contributes to the advancement of economic development. Urbanization, industrialization, and economic growth have enhanced living conditions, promoting sedentary behaviours among individuals with elevated socio-economic statuses. The prevalence of overweight and obesity has increased globally, along with the rise in living standards, posing a growing health risk. Overweight and obesity are known to be precursors to a range of modifiable and metabolic risk factors, contributing to the increase in lifestyle diseases. Unhealthy dietary choices and lifestyle practices primarily drive these diseases. Lifestyle diseases refer to chronic and non-communicable illnesses, encompassing conditions such as hypertension, heart disease, kidney disorders, infertility, polycystic ovary syndrome (PCOD), diabetes, respiratory diseases, and certain forms of cancer. This study examines the prevalence of lifestyle diseases among urban Indian women, explicitly differentiating between individuals with normal weight and those classified as obese. The main aim of this study is to investigate whether there is a correlation between a higher body mass index (BMI) in women and an increased vulnerability to lifestyle diseases when compared to women with a normal BMI. This research provides insights into the intricate relationship between lifestyle, health, and economic development within urban areas.

Keywords: urbanization, economic development, BMI, NCDs, urban, women, lifestyle practices

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Authors: Ria Forner

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Introduction: Due to increases in the SNR at 7T and above, it becomes more favourable to make use of X-nuclear imaging. Integrated body coils tuned to 120MHz for 31P, 79MHz for 23Na, and 75 MHz for 13C at 7T were simulated with a human male, female, or child body model to assess strategies of use for metabolic MR imaging in the body. Methods: B1 and SAR efficiencies in the heart, liver, spleen, and kidneys were assessed using numerical simulations over the three frequencies with phase shimming. Results: B1+ efficiency is highly variable over the different organs, particularly for the highest frequency; however, local SAR efficiency remains relatively constant over the frequencies in all subjects. Although the optimal phase settings vary, one generic phase setting can be identified for each frequency at which the penalty in B1+ is at a max of 10%. Discussion: The simulations provide practical strategies for power optimization, B1 management, and maintaining safety. As expected, the B1 field is similar at 75MHz and 79MHz, but reduced at 120MHz. However, the B1 remains relatively constant when normalised by the square root of the peak local SAR. This is in contradiction to generalized SAR considerations of 1H MRI at different field strengths, which is defined by global SAR instead. Conclusion: Although the B1 decreases with frequency, SAR efficiency remains constant throughout the investigated frequency range. It is possible to shim the body coil to obtain a maximum of 10% extra B1+ in a specific organ in a body when compared to a generic setting.

Keywords: birdcage, multi-nuclear, B1 shimming, 7 Tesla MRI, liver, kidneys, heart, spleen

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