Search results for: glucose metabolism

Authors: Nadeem Bashir, Ghulam Mustafa Peerzada

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The present study pertains to the nonlinear behavior of acetaminophen based uncatalyzed Belousov-Zhabotinsky (BZ) oscillator and its dynamics in the presence of Ferroin as the catalyst. The role of free metal ions as catalysts was examined and the results compared with corresponding complexed catalysts. Free metal ions were found to be sluggish with respect to the evolution of the oscillatory regime as compared to complexed ones. Effect of change of the ligand moiety of the catalyst complex on the oscillatory parameters was monitored. Since ethanol potentiates the hepatotoxicity caused by acetaminophen in-vivo, it is thought to understand this interaction by virtue of causing perturbation of the acetaminophen based oscillator with different concentrations of the ethanol with and without ferroin as the catalyst. Another dimension to the ethanol effect was added by perturbation of the system with ethanol at different stages of the reaction so as to get an idea whether it is acetaminophen or some reactive intermediate generated in the reaction system which reacts with ethanol. Further, the ferroin-catalyzed oscillator is taken as a prototype inorganic model of the acetaminophen-ethanol syndrome, as ferroin and HOBr were inorganic replacements to Cyt P450 and NADPH in the alcohol metabolism.

Keywords: Belousov-Zhabotinsky reaction, ferroin, Paracetamol-Ethanol syndrome, kinetics

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Authors: Haralabos C. karantonis, Afroditi Michalaki

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High intakes of carbohydrates and fats have been associated with an increased risk of chronic diseases due to the role of postprandial oxidative stress. This pilot nutritional intervention aimed to examine the acute effect of consuming two different types of breadsticks prepared from durum wheat flour mixtures differing in total phenolic content on postprandial inflammatory and oxidant responses in healthy volunteers. A cross-over, controlled, and single-blind clinical trial was designed, and two isocaloric high-fat and high-carbohydrate meals were tested. Serum total, High Density Lipoprotein (HDL)- and Low Density Lipoprotein (LDL)-cholesterol, triglycerides, glucose, C-reactive protein (CRP), uric acid, plasma total antioxidant capacity, and antiplatelet activity were determined in fasting and 30, 60, and 120 min after consumption. The results showed a better postprandial HDL-cholesterol and total antioxidant activity response in the intervention group. The choice of durum wheat flours with higher phenolic content and antioxidant activity is presented as promising for human health, and clinical studies will expand to draw safer conclusions.

Keywords: antioxidant, antiplatelet, durum wheat, nutritional value

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Authors: Afshin Farahbakhsh, Hoda Khodadadi

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In this paper, we compare five methods of biological fuel cell fabrication by combining a Shewanella oneidensis microbial anode and a laccase-modified air-breathing cathode. As a result of biofuel cell laccase with graphite nanofibers, carbon surface (PAMAN) on the pt/hpg electrode, graphite sheets MWCNT and with (PG) and (MWCNT) showed, respectively. Describes methods for creating controllable and reproducible bio-anodes and demonstrates the versatility of hybrid biological fuel cells. The laccase-based biocathodes prepared either with the crude extract or with the purified enzyme can provide electrochemically active and stable biomaterials. The laccase-based biocathodes prepared either with the crude extract or with the purified enzyme can provide electrochemically active and stable biomaterials. When the device was fed with transdermal extracts, containing only 30μM of glucose, the average peak power was proportionally lower (0.004mW). The result of biofuel cell with graphite nanofibers showed the enzymatic fuel cell reaches 0.5 V at open circuit voltage with both, ethanol and methanol and the maximum current density observed for E2electrode was 228.94mAcm.

Keywords: enzymatic electrode, fuel cell, immobilization, laccase

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Authors: Huong M. T. Nguyen, Hoa A. P. Nguyen, Chi V. Phan, Mai P. T. Nguyen, Ngoc D. Ngo, Van T. Ta, Hai T. Le

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Wilson’s disease is an autosomal recessive disorder of copper metabolism, which is caused by mutation in copper- transporting P-type ATPase (ATP7B). The mechanism of this disease is a failure of hepatic excretion of copper to the bile, and it leads to copper deposits in the liver and other organs. Most clinical symptoms of Wilson’s disease can present as liver disease and/or neurologic disease. Objective: The goal of the study is prenatal diagnosis for pregnant women at high risk of Wilson’s disease in Northern Vietnam. Material and method: Three probands with clinically diagnosed liver disease were detected in the mutations of 21 exons and exon-intron boundaries of the ATP7B gene by direct Sanger-sequencing. Prenatal diagnoses were performed by amniotic fluid sampling from pregnant women in the 16th-18th weeks of pregnancy after the genotypes of parents with the probands were identified. Result: A total of three different mutations of the probands, including of S105*, P1052L, P1273G, were detected. Among three fetuses which underwent prenatal genetic testing, one fetus was homozygote; two fetuses were carriers. Conclusion: Genetic testing provided a useful method for prenatal diagnosis, and is a basis for genetic counseling.

Keywords: ATP7B gene, genetic testing, prenatal diagnosis, pedigree, Wilson disease

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Authors: Amal Baalash, Shazia Mukaddam, M. Adel El-Sayed

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Background: Several studies have suggested non-skeletal effects of vitamin D and linked its deficiency with features of many chronic conditions. In this study, We aimed to investigate the relationship between vitamin D levels and different components of the metabolic syndrome in male and female Saudi patients. Methods: the study population consisted of 111 patients with metabolic syndrome (71 females and 40 males) aged 37-63 years enrolled from patients attending the internal medicine outpatient clinics of King Fahad Medical City. The parameters for diagnosis of the metabolic syndrome according to the National Cholesterol Education Program Adult Treatment Panel III (NCEP ATP III) were measured, which included waist circumference, TG, HDL-C, Blood pressure and fasting blood glucose (FBS). The association between each parameter and serum 25-hydroxyvitamin D (25(OH) D) was studied in both male and female patients separately. Results: in male patients, 25(OH) D levels were inversely associated with FBS and TG and positively associated with HDL-C and diastolic blood pressure, With highest association with the HDL-C levels. On the other hand 25(OH) D, Showed no significant association with any of the measured metabolic syndrome parameters in female patients. Conclusion: in Saudi patients with metabolic syndrome, the association between the parameters of metabolic syndrome and the levels of 25 (OH) D is more pronounced in males rather than females.

Keywords: gender, metabolic syndrome, Saudi patients, vitamin D

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Authors: Kiran Fatima, Kashif Ali

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Staphylococcus aureus is an opportunistic human as well as animal pathogen that causes a variety of diseases. A total of 70 staphylococci isolates were obtained from soil, water, yogurt, and clinical samples. The likely staphylococci clinical isolates were identified phenotypically by different biochemical tests. Molecular identification was done by PCR using species-specific 16S rRNA primer pairs, and finally, 50 isolates were found to be positive as Staphylococcus aureus, sciuri, xylous and cohnii. Screened isolates were further analyzed by several microbiological diagnostics tests, including gram staining, coagulase, capsule, hemolysis, fermentation of glucose, lactose, maltose, and sucrose tests enzymatic reactions. It was found that 78%, 81%, and 51% of isolates were positive for gelatin hydrolysis, protease, and lipase activities, respectively. Antibiogram analysis of isolated Staphylococcus aureus strains with respect to different antimicrobial agents revealed resistance patterns ranging from 57 to 96%. Our study also shows 70% of strains to be MRSA, 54.3% as VRSA, and 54.3% as both MRSA and VRSA. All the identified isolates were subjected to detection of mecA, nuc, and hlb genes, and 70%, 84%, and 40% were found to harbour mecA, nuc, and hlb genes, respectively. The current investigation is highly important and informative for the high-level multidrug-resistant Staphylococcus aureus infections inclusive also of methicillin and vancomycin.

Keywords: MRSA, VRSA, mecA, MSSA

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Authors: Bensaid Fatiha

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The objective of our present work is the description of the antagonistic capacities of one strain of Pseudomonas fluorescens and the nonpathogenic fungic isolate Fusarium oxysporum against phytopathogenic agent Fusarium oxysporum F. Sp. lycopersici. This work has been achieved in two main parts: the first is interested on the in vitro antagonistic activities; the second was interested to study the soil receptiveness of fusarium wilt tomato. The use of strain of fluorescent Pseudomonas and a non-pathogenic strain of F. oxysporum in the different antagonism tests, has allowed assuring a certain bio-protection from the plants of tomatoes opposite to F. oxysporum F. Sp. lycopersici, agent of a wilt of tomato. These antagonistic have shown a substantial in vitro antagonistic activity on the three mediums (KB, PDA, KB+PDA) against F. oxysporum F. Sp. lycopersici, by inhibiting its growth mycelium with rate of inhibition going until 80 % with non-pathogen of Fusarium oxysporum and 60 % with strain of fluorescens Pseudomonas. Soil microbial balance, between the antagonistic population and that of pathogenic, can be modulated through microbiological variations or abiotic additives influencing directly or indirectly the metabolic behavior microbial. In this experiment, addition of glucose or EDTA, could increase or decrease the resistance of soil by activation of pathogenic or antagonists, as a result of modification and modulation in their metabolic activities.

Keywords: fluorescents, nonpathogenic, fusarium oxysporum, fusarium wilt, antagonism, biological control, soil receptivity

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Authors: Yao-Te Yen, Chao-Hsin Cheng, Meng-Shun Huang, Shan-Zong Cyue

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'Poppy-seed defense' has been a serious problem all over the world, that is because the opiates metabolites in urine are difficult to distinguish where they come from precisely. In this research, a powerful analytic method has been developed to trace the opiates metabolites in urine by Gas-Chromatography Isotope Ratio Mass Spectrometry (GC-IRMS). In order to eliminate the interference of synthesis to heroin or metabolism through human body, opiates metabolites in urine and sized heroin were hydrolyzed to morphine. Morphine is the key compound for tracing between opiates metabolites and seized heroin in this research. By matching δ13C and δ15N values through morphine, it is successful to distinguish the opiates metabolites coming from heroin or medicine. We tested seven heroin abuser’s metabolites and seized heroin in crime sites, the result showed that opiates metabolites coming from seized heroin, the variation of δ13C and δ15N for morphine are within 0.2 and 2.5‰, respectively. The variation of δ13C and δ15N for morphine are reasonable with the result of matrix match experiments. Above all, the uncertainty of 'Poppy-seed defense' can be solved easily by this analytic method, it provides the direct evidence for judge to make accurate conviction without hesitation.

Keywords: poppy-seed defense, heroin, opiates metabolites, isotope ratio mass spectrometry

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Authors: Z. M. Iqbal, M. Abdullah, K. Javed, M. A. Jabbar, A. Haque, M. Saadullah, F. Shahzad

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The current study was conducted to set the appropriate concentrate level for Nili-Ravi buffalo heifers. Twenty seven buffalo heifers were randomly divided into three different groups A, B and C having nine animals in each group. All the heifers were given free access to chopped green fodder and fresh water. In addition, heifers of group A, B and C were given concentrate at the rate of 0.5%, 1% and 1.5% of their body weight. The average daily dry matter intake was 2.69, 3.06 and 3.83 kg with average daily gain of 456.09, 398.56 and 515.87 gm in group A, B and C, respectively. The feed conversion ratio of heifers of these groups was 5.89, 7.74 and 7.52, respectively. There was non-significant (P>0.05) difference in the body measurements (height at wither, body length and heart girth), final body condition and scoring and blood serum (glucose, total protein and cholesterol) of heifers of all the three groups. The results of current study shows that there is non-significant (P>0.05) difference in the growth rate of Nili-Ravi heifers at varying levels of concentrate so, it is cost effective to raise 6-8 month calves by offering concentrate at the rate of 0.5% body weight along with free access of green fodder.

Keywords: concentrate level, buffalo heifer, body measurement, green fodder

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Authors: Khaled S. Al Salhen

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Aldehyde oxidase is molybdo-flavoenzyme involved in the oxidation of hundreds of endogenous and exogenous and N-heterocyclic compounds and environmental pollutants. Uncharged N-heterocyclic aromatic compounds such phenanthridine are commonly distributed pollutants in soil, air, sediments, surface water and groundwater, and in animal and plant tissues. Phenanthridine as uncharged N-heterocyclic aromatic compound was incubated with partially purified aldehyde oxidase from rainbow trout fish liver. Reversed-phase HLPC method was used to separate the oxidation products from phenanthridine and the metabolite was identified. The 6(5H)-phenanthridinone was identified the major metabolite by partially purified aldehyde oxidase from fish liver. Kinetic constant for the oxidation reactions were determined spectrophotometrically and showed that this substrate has a good affinity (Km = 78 ± 7.6 µM) for hepatic aldehyde oxidase, coupled with a relatively high oxidation rate (0.77± 0.03 nmol/min/mg protein). In addition, the kinetic parameters of hepatic fish aldehyde oxidase towards the phenanthridine substrate indicate that in vitro biotransformation by hepatic fish aldehyde oxidase will be a significant pathway. This study confirms that partially purified aldehyde oxidase from fish liver is indeed the enzyme responsible for the in vitro production 6(5H)-phenanthridinone metabolite as it is a major metabolite by mammalian aldehyde oxidase.

Keywords: aldehyde oxidase, fish, phenanthridine, specificity

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Authors: Nathalia Florencia Barros Azeredo, Josué Martins Gonçalves, Pamela De Oliveira Rossini, Koiti Araki, Lucio Angnes

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This work demonstrates the electroanalysis of uric acid (UA) at very low working potential (0 V vs Ag/AgCl) directly in body fluids such as saliva and sweat using electrodes modified with mixed -Ni0.75Zn0.25(OH)2 nanoparticles exhibiting stable electrocatalytic responses from alkaline down to weakly acidic media (pH 14 to 3 range). These materials were prepared for the first time and fully characterized by TEM, XRD, and spectroscopic techniques. The electrochemical properties of the modified electrodes were evaluated in a fast and simple procedure for uric acid analyses based on cyclic voltammetry and chronoamperometry, pushing down the detection and quantification limits (respectively of 2.3*10-8 and 7.6*10-8 mol L-1) with good repeatability (RSD = 3.2% for 30 successive analyses pH 14). Finally, the possibility of real application was demonstrated upon realization of unexpectedly robust and sensitive modified FTO (fluorine doped tin oxide) glass and screen-printed sensors for measurement of uric acid directly in real saliva and sweat samples, with no significant interference of usual concentrations of ascorbic acid, acetaminophen, lactate and glucose present in those body fluids (Fig. 1).

Keywords: nickel hydroxide, mixed catalyst, uric acid sensors, biofluids

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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: Nafisa Komilova, Plamena Angelova, Andrey Abramov, Ulugbek Mirkhodjaev

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Parkinson’s disease (PD) is a progressive neurodegenerative disorder which is induced by the loss of dopaminergic neurons in the midbrain. The mechanism of neurodegeneration is associated with the aggregation of misfolded proteins, oxidative stress, and mitochondrial disfunction. Considering this, the process of removal of unwanted organelles or proteins by autophagy is vitally important in neurons, and activation of these processes could be protective in PD. Short-time acidification of cytosol can activate mitophagy and autophagy, and here we used sodium pyruvate and sodium lactate in human fibroblasts with PD mutations (Pink1, Pink1/Park2, α-syn triplication, A53T) to induce changes in intracellular pH. We have found that both lactate and pyruvate in millimolar concentrations can induce short-time acidification of cytosol in these cells. It induced activation of mitophagy and autophagy in control and PD fibroblasts and protected against cell death. Importantly, the application of lactate to acute brain slices of control and Pink1 knockout mice also induced a reduction of pH in neurons and astrocytes that increase the level of mitophagy. Thus, acidification of cytosol by compounds which play important role in cell metabolism also can activate mitophagy and autophagy and protect cells in the familial form of PD.

Keywords: Parkinson's disease, mutations, mitophagy, autophagy

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Authors: Sdiqeh Jalali, M. R. Khazdair

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Liver X receptors (LXR) have an essential role in the regulation of cholesterol metabolism, and their activation increase ABCG5 and ABCG8 genes expression for the improvement of cholesterol excretion from the body during reverse cholesterol transport (RCT). The aim of this study was to investigate the effects of high-intensity interval (HIT) and low intensity continuous (LIT) trainings on gene expression of these substances after a high-fat diet in Wistar rats. Materials and Methods: Fifteen male Wistar rats were divided into 3 groups: control group (n = 5), HIT exercise group (n = 5) and LIT exercise group (n = 5). All groups used a high-fat diet for 13 weeks, and the HIT and LIT groups performed the specific training program. The expression of LXRβ, ABCG5, and ABCG8 genes was measured after the training period. Findings: Data analysis showed significantly higher levels of LXRβ, ABCG5, and ABCG8 gene expression in HIT and LIT groups compared to the control group (P ≤ 0.05). Conclusion: HIT and LIT trainings after a high-fat diet have beneficial effects on RCT that prevent heart attack. Also, HIT training may have a greater effect on cholesterol excretion during the reverse cholesterol transport mechanism than LIT.

Keywords: liver X receptor, atherosclerosis, interval training, endurance training

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Authors: Zi-Han Li, Lu Fang, Liang Wu, Dong-Yuan Chang, Manyuan Dong, Liang Ji, Qi Zhang, Ming-Hui Zhao, Sydney C.W. Tang, Lemin Zheng, Min Chen

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Uncoupling of mitochondrial respiration by chemical uncouplers has proven effective in ameliorating obesity, insulin resistance, and hyperglycemia, which were risk factors for diabetic nephropathy (DN). Recently, it was found that annexin A1(ANXA1) could improve mitochondrial function to mitigate DN progression. However, the underlying mechanism is not fully clear yet. Here, it was identified that uncoupling protein 1 (UCP1), an inner membrane protein of mitochondria, as a key to mitochondrial homeostasis improved by ANXA1. Specifically, ANXA1 attenuated mitochondrial dysfunction via appropriately upregulating UCP1 by stabilizing its transcription factor GATA binding protein 3 (GATA3) through combining with thioredoxin. Moreover, specific overexpression of UCP1 in renal cortex rescued renal injuries in diabetic Anxa1-KO mice. UCP1 deletion aggravated renal injuries in HFD/STZ-induced diabetic mice. Mechanistically, UCP1 reduced mitochondrial fission through the aristaless-related homeobox (ARX)/cardiolipin synthase 1 (CRLS1) pathway. Therapeutically, CL316243, a UCP1 agonist, could attenuate established DN in db/db mice. This work established a novel principle to harness the power of uncouplers for the treatment of DN.

Keywords: diabetic nephropathy, uncoupling protein 1, mitochondrial homeostasis, cardiolipin metabolism

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Authors: Adele Broodryk, Cindy Pienaar, Martinique Sparks, Ben Coetzee

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Background: Cold water immersion (CWI) is a popular recovery modality utilised. However, discrepancies exist regarding the results over a 48 hour recovery period. Aim: To evaluate the effects of CWI and passive recovery (PAR) on a range of haematological and physical parameters over a 48-hour using a cross-sectional, pre-post-test design. Subjects and Methods: Both the and physical parameters were evaluated at baseline, after a 15-min fitness session, and at 0, 24 and 48 hours post-recovery in 23 male university rugby players. The CWI group sat in a cold water pool (8°C) for 20 min whereas the PAR group remained seated. Results: At 0 hours post-CWI, three (blood lactate (BLa-), Sodium (Na+) and haemoglobin) returned to baseline values, however Vertical Jump Test (VJT) height results decreased whereas after PAR it improved. From 0 to 24 and/or 48 h, four (Partial Oxygen (PO2) VJT-height, plasma glucose, and Na+) significantly increased (p ≤ 0.05) in either and/or both groups. Significant intergroup differences (p ≤ 0.05) were noticed in the physical tests. Conclusions: PAR is superior as an acute modality (0 hours) due to CWI cooling the body down. However, CWI demonstrates advantageous over a 24-hour period in a wide range of haematological variables.

Keywords: cryotherapy, recuperation, haematological, rugby

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Authors: Barbora Chmelova, Radek Sachl

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Gangliosides are amphipathic membrane lipids. Due to the composition of bulky oligosaccharide chains containing one or more sialic acids linked to the hydrophobic ceramide base, gangliosides are classified among glycosphingolipids. This unique structure induces a high self-aggregating tendency of gangliosides and, therefore, the formation of nanoscopic clusters called nanodomains. Gangliosides are preferentially present in an extracellular membrane leaflet of all human tissues and thus have an impact on a huge number of biological processes, such as intercellular communication, cell signalling, membrane trafficking, and regulation of receptor activity. Defects in their metabolism, impairment of proper ganglioside function, or changes in their organization lead to serious health conditions such as Alzheimer´s and Parkinson´s diseases, autoimmune diseases, tumour growth, etc. This work mainly focuses on ganglioside organization into nanodomains and their dynamics within the plasma membrane. Current research investigates static ganglioside nanodomains characterization; nevertheless, the information about their diffusion is missing. In our study, fluorescence correlation spectroscopy is implemented together with stimulated emission depletion (STED-FCS), which combines the diffraction-unlimited spatial resolution with high temporal resolution. By comparison of the experiments performed on model vesicles containing 4 % of either GM1, GM2, or GM3 and Monte Carlo simulations of diffusion on the plasma membrane, the description of ganglioside clustering, diffusion of nanodomains, and even diffusion of ganglioside molecules inside investigated nanodomains are described.

Keywords: gangliosides, nanodomains, STED-FCS, flourescence microscopy, membrane diffusion

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Authors: A. M. Almujalli, G. M. Al-Ghamdi

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Mycobacterium avium subspecies paratuberculosis causes paratuberculosis, a chronic debilitating granulomatous enteritis, in camels as well as domestic and wild ruminants. The clinical manifestation of the disease in camel is not well characterized, therefore this study was aimed to investigate the clinical and pathological pictures of camels that are suffering from partuberculosis. Twelve young camels that were presented to the Veterinary Teaching Hospital, King Faisal University were investigated. Clinical and pathological examination were performed. The results revealed highly significant increase in creatinine, blood urea nitrogen, magnesium, AST and ALT in diseased camels, while glucose, total protein and albumin were highly significantly decreased in diseased camels when compared to healthy ones. Post-mortem testing indicated thickening, corrugation of the intestinal wall, folded mucosa, enlarged and oedemated ileocaecal and mesenteric lymph nodes. The microscopic findings detected short, blunt and distorted intestinal villi with hyperactive goblet cells of the villi and the crypts of lieberkuhn contained mucin droplets. The lamina propria was heavily infiltrated with mononuclear cells mostly macrophages. This clinical picture of paratuberculosis may be used to initiate control strategy to limit the spread of the disease in camel herds.

Keywords: camel, partuberclosis, control, Saudi Arabia

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Authors: Saroj Poudel, Joshua Mancini, Douglas Pike, Jennifer Timm, Alexei Tyryshkin, Vikas Nanda, Paul Falkowski

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On the early Earth, protein-metal complexes likely harvested energy from a reduced environment. These complexes would have been precursors to the metabolic enzymes of ancient organisms. Hydrogenase is an essential enzyme in most anaerobic organisms for the reduction and oxidation of hydrogen in the environment and is likely one of the earliest evolved enzymes. To attempt to reinvent a precursor to modern hydrogenase, we computationally designed a short thirteen amino acid peptide that binds the often-required catalytic transition metal Nickel in hydrogenase. This simple complex can achieve hundreds of hydrogen evolution cycles using light energy in a broad range of temperature and pH. Biophysical and structural investigations strongly indicate the peptide forms a di-nickel active site analogous to Acetyl-CoA synthase, an ancient protein central to carbon reduction in the Wood-Ljungdahl pathway and capable of hydrogen evolution. This work demonstrates that prior to the complex evolution of multidomain enzymes, early peptide-metal complexes could have catalyzed energy transfer from the environment on the early Earth and enabled the evolution of modern metabolism

Keywords: hydrogenase, prebiotic enzyme, metalloenzyme, computational design

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Authors: Li-Yu Lee, Yu-Shuan Chen, Jun Sheng Wang, I-Ming Chu

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Microneedle patch is a painless transdermal drug delivery method, It could solve some problems in traditional drug delivery such as digestive system causing drug metabolism and subcutaneous injection causing some side effects. Coating drug on or loading drug in microneedle can carry active ingredient through stratum corneum, also can control dose well when microneedle patch apply on localized topical area. We used hyaluronic acid to fabricate dissolvable microneedle patch and encapsulated minoxidil into microneedles. Minoxdil is a drug for exterior use that can be used to treat Androgenetic alopecia, but related commercial products have some shortcomings, for example, propylene glycol which is used to soften stratum corneum cause skin allergic reaction, comparing chemical promotion, microneedle patch provide physical way to make drugs through nature barrier of skin. In this research, we designed a two-step process to fabricate microneedle patch, that can effectively reduce drug waste, and gentle production process could maintain drug activity well. We also do in vitro test on cadaver to make sure patch has enough mechanical strength to penetrate stratum corneum. In the release test and animal test, we found microneedle patch has higher delivery efficiency than tradition way. In this study, we may determine that germinal MNs patch is a potential commodity.

Keywords: dissolving microneedles, androgenetic alopecia, minoxidil, transdermal drug delivery

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Authors: Jinhyun Ryu, Chengliang Xie, Nal Ae Yoon, Dong Hoon Lee, Gu Seob Roh, Hyun Joon Kim, Gyeong Jae Cho, Wan Sung Choi, Sang Soo Kang

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Type 2 diabetes is a heterogeneous group of metabolic disorders featured by a deficit in or loss of insulin activity to maintain normal glucose homeostasis. Mainly, it results from the compromised insulin secretion and/or reduced insulin activity. The frequency of type 2 diabetes (T2D) has been increased rapidly in recent decades with the increase in the trend of obesity due to life style and food habit. Obesity is considered to be the primary risk factor for the development of insulin resistance and thereby developing T2D. Traditionally naturally occurring fruits, vegetables etc. are being used to treat many pathogenic conditions. In this study, we tried to find out the effect of a popularly used vegetable in Bangladesh and several other Asian countries, ‘bitter melon’ on high fat diet induced T2D. To investigate the effect, we used 70% ethanol extract of bitter melon (BME) as dietary supplement with chow. BME was found to attenuate the high fat diet (HFD) induced body weight and total fat mass significantly. We also observed that BME reduced the insulin resistance induced by HFD effectively. Furthermore, dietary supplementation of BME was highly effective in increasing insulin sensitivity, and reducing the hepatic fat and obesity. These results indicate that BME could be effective to attenuate T2D and could be a preventive measure against T2D.

Keywords: bitter melon, obesity, type 2 diabetes, high fat diet

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Authors: So Youn Park, Yi Sle Lee, Ki Whan Hong, Chi Dae Kim

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The role of metabolism in the pathogenesis of osteoarthritis is an emerging field. Metabolic alterations may be a role in osteoarthritis (OA) pathogenesis, and these changes influence joint destruction via several cytokine. Especially, in OA patients, levels of IL-1β are elevated in the synovial fluid, synovial membrane, subchondral bone, and cartilage. The IL-1β is activated by NLRP3 inflammasomes, and NLRP3 inflammasomes are cytosolic complexes that drive the production of other inflammatory cytokines, including IL-1β. In this study, we examined that SIRT1 suppresses IL-1β through inhibiting NLRP3 inflammasomes and SIRT1 ameliorates osteoarthritis. OA fibroblasts were isolated from synovium of OA patients. IL-1β and NLRP3 were detected in synovium of OA patients by immunohistochemistry. Lipopolysaccharides (LPS) stimulated the expression of active IL-1β mRNA in OA fibroblasts and combination of LPS, and adenosine triphosphate increased more the expression of active IL-1β in OA fibroblasts. The level of IL-1β was measured by western blot and ELISA assay. NLRP3 inflammasomes complex were measured by western blot. SIRT1 did not inhibit expression of NLRP3 inflammasome. So caspase-1, apoptotic speck-like protein containing a caspase recruitment domain (ASC) and NLRP3 protein were expressed in OA fibroblasts. But SIRT1 suppressed activation of IL-1β by inhibiting activity of caspase-1 via NLRP3 inflammasome in OA fibroblasts under LPS plus ATP stimulation. These results suggest that SIRT1 is a modulator of NLRP3 inflammasomes in OA fibroblasts and ameliorate IL-1β, so expression of SIRT1 in OA fibroblast may be a potential strategy for OA inflammation treatment.

Keywords: osteoarthritis, inflammasome, SIRT1, IL-1beta

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Authors: Sanaa Bardaweel

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Inorganic polyphosphate (poly P) is present in all living forms tested to date, from each of the three kingdoms of life. Studied mainly in prokaryotes, poly P and its associated enzymes are vital in diverse basic metabolism, in at least some structural functions and, notably, in stress responses. These plentiful and unrelated roles for poly P are probably the consequence of its presence in life-forms early in evolution. The genomes of many bacterial species, including pathogens, encode a homologue of a major poly P synthetic enzyme, poly P kinase 1 (PPK1). Genetic deletion of ppk1 results in reduced poly P levels and loss of pathogens virulence towards protozoa and animals. Thus far, no PPK1 homologue has been identified in higher-order eukaryotes and, therefore, PPK1 represents a novel target for chemotherapy. The idea of the current study is to purify the PPK1 from Escherichia coli to homogeneity in order to study the effect of active site point mutations on PPK1 catalysis via the application of site-directed mutagenesis strategy. The knowledge obtained about the active site of PPK1 will be utilized to characterize the catalytic and kinetic mechanism of PPK1 with model substrates. Comprehensive understanding of the enzyme kinetic mechanism and catalysis will be used to design and screen a library of synthetic compounds for potential discovery of selective PPK1-inhibitors.

Keywords: antimicobial, Escherichia coli, inorganic polyphosphate, PPK1-inhibitors

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Authors: Musarat Ishaq, Tara Karnezis, Ramin Shayan

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Lipedema, a poorly understood chronic disease of adipose hyper-deposition, is often mistaken for obesity and causes significant impairment to mobility and quality-of-life. To identify molecular mechanisms underpinning lipedema, we employed comprehensive omics-based comparative analyses of whole tissue, adipocyte precursors (adipose-derived stem cells (ADSCs)), and adipocytes from patients with or without lipedema. Transcriptional profiling revealed significant differences in lipedema tissue, adipocytes, and ADSCs, with altered levels of mRNAs involved inproliferation and cell adhesion. One highly up-regulated gene in lipedema adipose tissue, adipocytes and ADSCs, ZIC4, encodes Zinc Finger Protein ZIC 4, a class of transcription factor which may be involved in regulating metabolism and adipogenesis. ZIC4 inhibition impaired the adipogenesis of ADSCs into mature adipocytes. Epigenetic regulation study revealed overexpression of ZIC4 is involved in decreased promoter DNA methylation and subsequent decrease in adipogenesis. These epigenetic modifications can alter adipocytes microenvironment and adipocytes differentiation. Our study show that epigenetic events regulate the ability of ADSCs to commit and differentiate into mature adipocytes by modulating ZIC4.

Keywords: lipedema, adipose-derived stem cells, adipose tisue, adipocytes, zinc finger protein, epigenetic

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Authors: Niranjan Koirala, Sumangala Darsandhari, Hye Jin Jung, Jae Kyung Sohng

Abstract:

Sakuranetin, an antifungal agent and genkwanin, an anti-inflammatory agent, are flavonoids with several potential pharmaceutical applications. To produce such valuable flavonoids in large quantity, an Escherichia coli cell factory has been created. E. coli harboring O-methyltransferase (SaOMT2) derived from Streptomyces avermitilis was employed for regiospecific methylation of naringenin and apigenin. In order to increase the production via biotransformation, metK gene was overexpressed and the conditions were optimized. The maximum yield of sakuranetin and genkwanin under optimized conditions was 197 µM and 170 µM respectively when 200 µM of naringenin and apigenin were supplemented in the separate cultures. Furthermore, sakuranetin was purified in large scale and used as a substrate for in vitro glycosylation by YjiC to produce glucose and galactose derivatives of sakuranetin for improved solubility. We also found that unlike naringenin, sakuranetin effectively inhibits α-melanocyte stimulating hormone (α-MSH)-stimulated melanogenesis in B16F10 melanoma cells. In addition, genkwanin more potently inhibited angiogenesis than apigenin. Based on our findings, we speculate that these compounds warrant further investigation in vivo as potential new therapeutic anti-carcinogenic, anti-melanogenic and anti-angiogenic agents.

Keywords: anti-carcinogenic, anti-melanogenic, glycosylation, methylation

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Authors: Wisnu Adi Yulianto, Chatarina Lilis Suryani, Mamilisti Susiati, Hendy Indra Permana

Abstract:

Parboiled rice was developed to produce rice that has low glycemic index, especially for diabetics. Yet, parboiled rice is not enough because diabetics also lack of chromium. The sign of chromium (Cr) deficiency in diabetics is impaired glucose tolerance. Cr fortification was done for increasing Cr content in rice. Naturally-occurring compounds that have been proven to improve insulin sensitivity include Cr and polyphenol found in cinnamon, pandan and bay leaf. This research aimed to evaluate content of resistant starch and glycemic index of Cr - fortified - parboiled rice (Cr-PR) coated with herbal extracts. Variety of unhulled rice and forticant used in the experiment were Ciherang and CrCl3, respectively. Three herbal extracts used were cinnamon, pandan and bay leaf. Each concentration of herbal extracts in the amount of 3%, 6%, and 9% were added in the coating substance to coat Cr-PR. Resistant starch (RS) content was determined by enzymatic process through glucooxydase method. Testing of the GI was conducted on 18 non-diabetic volunteers. RS content of Cr-PR coated with herbal extracts ranged between 8.27 – 8.84 % (dry weight). Cr-PR coated with all herbal extracts of 3% concentration had higher RS content than the ones with herbal extracts of 6% and 9% concentration (P <0.05). Value of the rice GI ranged 29 - 40. The lowest GI (29-30) was attained by the rice coated with enrichment of 6-9% cinnamon extract.

Keywords: coating, Cr-fortified-parboiled rice, glycemic index, herbal extracts, resistant starch

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Authors: Mirnawati, G. Ciptaan, Ferawati

Abstract:

This study aims to improve the quality of the cassava peel-leaf mixture (CPLM) through fermentation with Rhizopus oligosporusas poultry ration. This research is an experimental study using a completely randomized design (CRD) with four treatments and five replications. The treatments were cassava peel-leaf mixture (CPLM) fermented with Rhizopus oligosporus. The treatments were a combination of cassava peel and leaves with the ratio of; A (9:1), B (8:2), C (7:3), and D (6:4). The observed variables were protease enzyme activity, crude protein, crude fiber, nitrogen retention, digestibility of crude fiber, and metabolic energy. The results of the diversity analysis showed that there was a very significant (p < 0.01) effect on protease activity, crude protein, crude fiber, nitrogen retention, digestibility of crude fiber, and energy metabolism of fermented CPLM. Based on the results of the study, it can be concluded that CPLM (6:4) fermented with Rhizopus oligosporus gave the best results seen from protease activity 7,25 U/ml, 21.23% crude protein, 19.80% crude fiber, 59.65% nitrogen retention, 62.99% crude fiber digestibility and metabolic energy 2671 Kcal/kg.

Keywords: quality, Casava peel-leaf mixture, fermentation, Rhizopus oligosporus

Procedia PDF Downloads 185

Authors: Li Bixia, Ren Shouwen, Fu Yanfeng, Tu Feng, Xiaoming Fang, Xueming Wang

Abstract:

The utilization of dietary crude fiber in different breed pigs is not the same. Su-shan pigs are a new breed formed by crossing Taihu pigs and Yorkshire pigs. In order to understand the resistance of Su-shan pigs to dietary crude fiber, 150 Su-shan pigs with 60 kg of average body weight and similar body conditions were allocated to three groups randomly, and there are 50 pigs in each group. The percentages of dietary crude fiber were 8.35%, 9.10%, and 11.39%, respectively. At the end of the experiment, 15 pigs randomly selected from each group were slaughtered. The results showed as follows: average daily gain of the 9.10% group was higher than that of the 8.35% group and the 11.39% group; there was a significant difference between the 9.10% group and the 8.35% group (p < 0.05. Levels of urea nitrogen, total cholesterol and high density lipoprotein in the 9.10% group were significantly higher than those in the 8.35% group and the 11.39% group (p < 0.05). Ratios of meat to fat in the 9.10% group and the 11.39% group were significantly higher than that in the 8.35% group (p < 0.05). Lean percentage of 9.10% group was higher than that of 8.35% group and 11.39% group, but there was no significant difference in three groups (p > 0.05). The weight of small intestine and large intestine in the 11.39% group was higher than that in the 8.35% group, and the 9.10% group and the difference reached a significant level (p < 0.05). In conclusion, increasing dietary crude fiber properly could reduce fat percentage, and improve the ratio of meat to fat of finishing Su-shan pigs. The digestion and metabolism of dietary crude fiber promoted the development of stomach and intestine of finishing Su-shan pig.

Keywords: Su-shan pigs, dietary crude fiber, growth performance, serum biochemical indexes

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Authors: Naso Isaiah Thanavisuth

Abstract:

Medical cannabis can be used for treatment including pain, multiple sclerosis, Parkinson's disease, and cancer. However, medical cannabis leads to adverse effects (AEs), which is delta-9-tetrahydrocannabinol (THC). In previous studies, the major of THC metabolism enzymes are CYP2C9. Especially, the variation of CYP2C9 gene consist of CYP2C9*2 on exon 3 and CYP2C9*3 on exon 7 to decrease enzyme activity. Notwithstanding, there is no data describing whether the variant of CYP2C9 genes are apharmacogenetics marker for the prediction of THC-induced AEs in Thai patients. We want to investigate the association between CYP2C9 gene and THC-induced AEs in Thai patients. We enrolled 39 Thai patients with medical cannabis treatment who were classified by clinical data. The CYP2C9*2 and *3 genotyping were conducted using the TaqMan real time PCR assay. All Thai patients who received the medical cannabis consist of twenty-four (61.54%) patients were female, and fifteen (38.46%) were male, with age range 27- 87 years. Moreover, the most AEs in Thai patients who were treated with medical cannabis between cases and controls were tachycardia, arrhythmia, dry mouth, and nausea. Particularly, thirteen (72.22%) medical cannabis-induced AEs were female and age range 33 – 69 years. In this study, none of the medical cannabis groups carried CYP2C9*2 variants in Thai patients. The CYP2C9*3 variants (*1/*3, intermediate metabolizer, IM) and (*3/*3, poor metabolizer, PM) were found, three of thirty-nine (7.69%) and one of thirty-nine (2.56%), respectively. Although, our results indicate that there is no found the CYP2C9*2. However, the variation of CYP2C9 allele might serve as a pharmacogenetics marker for screening before initiating the therapy with medical cannabis for the prevention of medical cannabis-induced AEs.

Keywords: CYP2C9, medical cannabis, adverse effects, THC, P450

Procedia PDF Downloads 119

Authors: Swapna Guntupalli, Leela Madhuri Chalasani, Kshatri Jyothi, C. V. Rao, Bondili J. S.

Abstract:

The study hypothesizes that enhanced biodegradation of Polycyclic Aromatic Hydrocarbons (PAHs) is achievable with an assemblage of microorganisms that are capable of producing biofilm and biosurfactants. Accordingly, PAHs degrading microorganism’s (bacteria, fungi, actinomycetes and yeast) were screened and grouped into different consortia based on their capabilities to produce biofilm and biosurfactants. Among these, Consortium BTSN09 consisting of bacterial fungal cocultures showed highest degradation due to the synergistic action between them. Degradation effiencies were evaluated using HPLC and GC-MS. Within 7days, BTSN09 showed 51% and 50.7% degradation of Phenanthrene (PHE) and Pyrene (PYR) with 200mg/L and 100 mg/L concentrations respectively in a liquid medium. In addition, several degradative enzymes like laccases, 1hydroxy-2-naphthoicacid dioxygenase, 2-carboxybenzaldehyde dehydrogenase, catechol1,2 dioxygenase and catechol2,3 dioxygenase activity was observed during degradation. Degradation metabolites were identified using GC-MS analysis and from the results it was confirmed that the metabolism of degradation proceeds via pthalic acid pathway for both PAHs. Besides, Microbial consortia also demonstrated good biosurfactant production capacity, achieving maximum oil displacement area and emulsification activity of 19.62 cm2, 65.5% in presence of PAHs as sole carbon source. Scanning Electron Microscopy analysis revealed exopolysaccharides (EPS) production, micro and macrocolonies formation with different stages of biofim development in presence of PAHs during degradation.

Keywords: PAHs, biosurfactant, biofilm, biodegradation

Procedia PDF Downloads 582