Search results for: protein metabolism

Authors: Maryam Beiranvand

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Multi-drug resistance in various microorganisms has increased globally in many healthcare facilities. Less effective antimicrobial activity of drug therapies for infection control becomes trouble. Since 1980, no new type of antimicrobial drug has been identified, even though combinations of antibiotic drugs have been discovered almost every decade. Between 1981 and 2006, over 70% of novel pharmaceuticals and chemical agents came from natural sources. Microorganisms have yielded almost 22,000 natural compounds. The identification of antimicrobial components from endophytes bacteria could help overcome the threat posed by multi-drug resistant strains. The project aims to analyze and identify antimicrobial peptides isolated from entophytic bacteria and their activity against multidrug-resistant Gram-negative bacteria in South Korea. Endophytic Paenibacillus polymyxa. 4G3 isolated from the plant, Gynura procumbery exhibited considerable antimicrobial activity against Methicillin-resistant Staphylococcus aureus, and Escherichia coli. The Rapid Annotations using Subsystems Technology showed that the total size of the draft genome was 5,739,603bp, containing 5178 genes with 45.8% G+C content. Genome annotation using antiSMASH version 6.0.0 was performed, which predicted the most common types of non-ribosomal peptide synthetase (NRPS) and polyketide synthase (PKS). In this study, diethyl aminoethyl cellulose (DEAEC) resin was used as the first step in purifying for unknown peptides, and then the target protein was identified using hydrophilic and hydrophobic solutions, optimal pH, and step-by-step tests for antimicrobial activity. This crude was subjected to C18 chromatography and elution with 0, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, and 100% methanol, respectively. Only the fraction eluted with 20% -60% methanol demonstrated good antimicrobial activity against MDR E. coli. The concentration of the active fragment was measured by the Brad-ford test, and Protein A280 - Thermo Fisher Scientific at the end by examining the SDS PAGE Resolving Gel, 10% Acrylamide and purity were confirmed. Our study showed that, based on the combined results of the analysis and purification. P polymyxa. 4G3 has a high potential exists for producing novel functions of polymyxin E and bacitracin against bacterial pathogens.

Keywords: endophytic bacteria, antimicrobial activity, antimicrobial peptide, whole genome sequencing analysis, multi -drug resistance gram negative bacteria

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Authors: Alebiosu Olugbenga Shadrak, Victor Victoria

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Honey bees exhibit preferences in their foraging behaviour on pollen and nectar for food and honey production, respectively. Melissopalynology is the study of pollen in honey and other honey products. Several work have been conducted on the palynological studies of honeys from the southern parts of Nigeria but with relatively scant records from the Northern region of the country. This present study aimed at revealing the favourably visited plants by honey bees, Apis melifera var. adansonii, at some apiaries in Northern Nigeria, as well as determining the quality of honeys produced. Honeys were harvested and collected from four apiaries of the region, namely: Sarkin Dawa missionary bee farm, Taraba State; Eleeshuwa Bee Farm, Keffi, Nassarawa State, Bulus Beekeeper Apiaries, Kagarko, Kaduna State and Mai Gwava Bee Farm, Kano State. These honeys were acetolysed for palynological microscopic analysis and subjected to standard treatment methods for the determination of their proximate composition and sugar profiling. Fresh anthers of two dominantly represented plants in the honeys were then collected for the quantification of their pollen protein contents, using the micro-kjeldhal procedure. A total of 30 pollen types were identified in the four honeys, and some of them were common to the honeys. A classification method for expressing pollen frequency class was employed: Senna cf. siamea, Terminalia cf. catappa, Mangifera indica, Parinari curatelifolia, Vitellaria paradoxa, Elaeis guineensis, Parkia biglobosa, Phyllantus muellerianus and Berlina Grandiflora, as “Frequent” (16-45%); while the others are either Rare (3-15%) or Sporadic (less than 3 %). Pollen protein levels of the two abundantly represented plants, Senna siamea (15.90mg/ml) and Terminalia catappa (17.33mg/ml) were found to be considerably lower. The biochemical analyses revealed varying amounts of proximate composition, non-reducing sugar and total sugar levels in the honeys. The results of this study indicate that pollen and nectar of the “Frequent” plants were preferentially foraged by honeybees in the apiaries. The estimated pollen protein contents of Senna same and Terminalia catappa were considerably lower and not likely to have influenced their favourable visitation by honeybees. However, a relatively higher representation of Senna cf. siamea in the pollen spectrum might have resulted from its characteristic brightly coloured and well scented flowers, aiding greater entomophily. Terminalia catappa, Mangifera indica, Elaeis guineensis, Vitellaria paradoxa, and Parkia biglobosa are typical food crops; hence they probably attracted the honeybees owing to the rich nutritional values of their fruits and seeds. Another possible reason for a greater entomophily of the favourably visited plants are certain nutritional constituents of their pollen and nectar, which were not investigated in this study. The nutritional composition of the honeys was observed to fall within the safe limits of international norms, as prescribed by Codex Alimentarius Commission, thus they are good honeys for human consumption. It is therefore imperative to adopt strategic conservation steps in ensuring that these favourably visited plants are protected from indiscriminate anthropogenic activities and also encourage apiarists in the country to establish their bee farms more proximally to the plants for optimal honey yield.

Keywords: honeybees, melissopalynology, preferentially foraged, nutritional, bee farms, proximally

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Authors: Asiri Wijesinghe, N. D. Kodikara, Damitha Sandaruwan

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The Diabetic Retinopathy (DR) is a rapidly growing interrogation around the world which can be annotated by abortive metabolism of glucose that causes long-term infection in human retina. This is one of the preliminary reason of visual impairment and blindness of adults. Information on retinal pathological mutation can be recognized using ocular fundus images. In this research, we are mainly focused on resurrecting an automated diagnosis system to detect DR anomalies such as severity level classification of DR patient (Non-proliferative Diabetic Retinopathy approach) and vessel tortuosity measurement of untwisted vessels to assessment of vessel anomalies (Proliferative Diabetic Retinopathy approach). Severity classification method is obtained better results according to the precision, recall, F-measure and accuracy (exceeds 94%) in all formats of cross validation. In ROC (Receiver Operating Characteristic) curves also visualized the higher AUC (Area Under Curve) percentage (exceeds 95%). User level evaluation of severity capturing is obtained higher accuracy (85%) result and fairly better values for each evaluation measurements. Untwisted vessel detection for tortuosity measurement also carried out the good results with respect to the sensitivity (85%), specificity (89%) and accuracy (87%).

Keywords: fundus image, exudates, microaneurisms, hemorrhages, tortuosity, diabetic retinopathy, optic disc, fovea

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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: Samantha A. Cintron, Janet Pierce, Mihaela E. Sardiu, Diane Mahoney, Jill Peltzer, Bhanu Gupta, Qiuhua Shen

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High output heart failure (HOHF) is characterized a high output state resulting from an underlying disease process and is commonly caused by obesity. As obesity levels increase, more individuals will be at risk for obesity-related HOHF. However, the underlying pathophysiologic mechanisms of obesity-related HOHF are not well understood and need further research. The aim of the study was to describe the differences in leukocyte transcriptomes of morbidly obese patients with HOHF and those with non-HOHF. In this cross-sectional study, the study team collected blood samples, demographics, and clinical data of six patients with morbid obesity and HOHF and six patients with morbid obesity and non-HOHF. The study team isolated the peripheral blood leukocyte RNA and applied stranded total RNA sequencing. Differential gene expression was calculated, and Ingenuity Pathway Analysis software was used to interpret the canonical pathways, functional changes, upstream regulators, and mechanistic and causal networks that were associated with the significantly different leukocyte transcriptomes. The study team identified 116 differentially expressed genes; 114 were upregulated, and 2 were downregulated in the HOHF group (Benjamini-Hochberg adjusted p-value ≤ 0.05 and log2(fold-change) of ±1). The differentially expressed genes were involved with cell proliferation, mitochondrial function, erythropoiesis, erythrocyte stability, and apoptosis. The top upregulated canonical pathways associated with differentially expressed genes were autophagy, adenosine monophosphate-activated protein kinase signaling, and senescence pathways. Upstream regulator GATA Binding Protein 1 (GATA1) and a network associated with nuclear factor kappa-light chain-enhancer of activated B cells (NF-kB) were also identified based on the different leukocyte transcriptomes of morbidly obese patients with HOHF and non-HOHF. To the author’s best knowledge, this is the first study that reported the differential gene expression in patients with obesity-related HOHF and demonstrated the unique pathophysiologic mechanisms underlying the disease. Further research is needed to determine the role of cellular function and maintenance, inflammation, and iron homeostasis in obesity-related HOHF.

Keywords: cardiac output, heart failure, obesity, transcriptomics

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Authors: William Kelly, Sorelle Veigne, Xianhua Li, Zuyi Huang, Shyamsundar Subramanian, Eugene Schaefer

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The ambr15™ bioreactor is a single-use microbioreactor for cell line development and process optimization. The ambr system offers fully automatic liquid handling with the possibility of fed-batch operation and automatic control of pH and oxygen delivery. With operating conditions for large scale biopharmaceutical production properly scaled down, micro bioreactors such as the ambr15™ can potentially be used to predict the effect of process changes such as modified media or different cell lines. In this study, gassing rates and dilution rates were varied for a semi-continuous cell culture system in the ambr15™ bioreactor. The corresponding changes to metabolite production and consumption, as well as cell growth rate and therapeutic protein production were measured. Conditions were identified in the ambr15™ bioreactor that produced metabolic shifts and specific metabolic and protein production rates also seen in the corresponding larger (5 liter) scale perfusion process. A Dynamic Flux Balance model was employed to understand and predict the metabolic changes observed. The DFB model-predicted trends observed experimentally, including lower specific glucose consumption when CO₂ was maintained at higher levels (i.e. 100 mm Hg) in the broth. A Computational Fluid Dynamic (CFD) model of the ambr15™ was also developed, to understand transfer of O₂ and CO₂ to the liquid. This CFD model predicted gas-liquid flow in the bioreactor using the ANSYS software. The two-phase flow equations were solved via an Eulerian method, with population balance equations tracking the size of the gas bubbles resulting from breakage and coalescence. Reasonable results were obtained in that the Carbon Dioxide mass transfer coefficient (kLa) and the air hold up increased with higher gas flow rate. Volume-averaged kLa values at 500 RPM increased as the gas flow rate was doubled and matched experimentally determined values. These results form a solid basis for optimizing the ambr15™, using both CFD and FBA modelling approaches together, for use in microscale simulations of larger scale cell culture processes.

Keywords: cell culture, computational fluid dynamics, dynamic flux balance analysis, microbioreactor

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Authors: S. Stankov, H. Fidan, N. Petkova, M. Stoyanova, Tz. Radoukova, A. Stoyanova

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The aim of this study was to evaluate the chemical composition of ripe Juniperus excelsa M. Bieb. galbuli (female cones) collected from "Izgoraloto Gyune" Reserve in Krichim, Bulgaria. The moisture (36.88%), abs. weight 693.96 g/1000 pcs., and the ash content (10.57%) of ripe galbuli were determined. Lipid fraction (9.12%), cellulose (13.54%), protein (13.64%), and total carbohydrates (31.20%) were evaluated in the ripe galbuli. It was found that the ripe galbuli contained glucose (4.00%) and fructose (4.25%), but disaccharide sucrose was not identified. The main macro elements presented in the sample were K (8390.00 mg/kg), Ca (4596.00 g/kg), Mg (837.72 mg/kg), followed by Na (7.69 mg/kg); while the detected microelements consisted of Zn (8.51 mg/kg), Cu (4.66 mg/kg), Mn (3.65 mg/kg), Fe (3.26 mg/kg), Cr (3.00 mg/kg), Cd (< 0.1 mg/kg), and Pb (0.01 mg/kg).

Keywords: chemical composition, Juniperus excelsa M. Bieb, minerals, ripe galbuli

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Authors: Valentin Nicorescu, Nicoleta C. Predescu, Camelia Papuc, Iuliana Gajaila, Carmen D. Petcu

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The oxidation of the fresh muscle oxymyoglobin (bright red colour) to metmyoglobin (brown colour) leads to discoloration of red meats. After slaughter, enzymatic systems involved in metmyoglobin reduction are continually depleted as time post-mortem progresses, thus the meat colour is affected. Phenolic compounds are able to scavenge reactive species involved in oxymyoglobin oxidation and to reduce metmyoglobin to oxymyoglobin. The aim of this study was to investigate the effect of polyphenols extracted from hawthorn fruits on the stability of oxymyoglobin and myofibrillar proteins in ground pork subject to refrigeration for 6 days. Hawthorn polyphenols (HP) were added in ground pork in 100, 200 and 300 ppm concentrations. Oxymyoglobin and metmyoglobin were evaluated spectrophotometrically at every 2 days and electrophoretic pattern of myofibrillar proteins was investigated at days 0 and 6 by Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE). For all meat samples, oxymyoglobin concentration significantly decreased during the first 4 days of refrigeration. After 6 days, the significant decrease of oxymyoglobin concentration continued only in the negative control samples. In samples treated with HP and butylated hydroxylanisole (BHA - positive control), oxymyoglobin concentration increased after 6 days of refrigeration, the highest levels complying with the following order: 100 ppm HP > 200 ppm HP > 300 ppm HP > 100 ppm BHA. The increase in metmyoglobin was coincidental with the decrease in oxymyoglobin; metmyoglobin concentration progressively increased during the first 4 days of refrigeration in all meat samples. After 6 days, in meat samples treated with HP and BHA, lower metmyoglobin concentrations were found (compared to day 4), respecting the following order: 100 ppm HP < 200 ppm HP < 300 ppm HP < 100 ppm BHA. These results showed that hawthorn polyphenols and BHA reduced metmyoglobin (MbFe3+) to oxymyoglobin (MbFe2+), and the strongest reducing character was recorded for 100 ppm HP. After 6 days of refrigeration, electrophoretic pattern of myofibrillar proteins showed minor changes compared to day 0, indicating that HP prevent protein degradation as well as synthetic antioxidant BHA. Also, HP did not induce cross-links in the myofibrillar proteins, to form protein aggregates, and no risk of reducing their ability to retain water was identified. The pattern of oxymyoglobin and metmyoglobin concentrations determined in this study showed that hawthorn polyphenols are able to reduce metmyoglobin to oxymyoglobin and to delay oxymyoglobin oxidation, especially when they are added to ground meat in concentration of 100 ppm. This work was carried out through Partnerships in priority areas Program – PN II, implemented with the support of MEN – UEFISCDI (Romania), project nr. 149/2014.

Keywords: Hawthorn polyphenols, metmyoglobin, oxymyoglobin, proteins stability

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Authors: Aulanni’am Aulanni’am

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Incidence of Diabetes Mellitus (DM) progressively increasing it became a serious problem in Indonesia and it is a disease that government is priority to be addressed. The longer a person is suffering from diabetes the more likely to develop complications particularly diabetic patients who are not well maintained. Therefore, Incidence of Diabetes Mellitus needs to be done in the early diagnosis of pre-phase of the disease. In this pre-phase disease, already happening destruction of pancreatic beta cells and declining in beta cell function and the sign autoimmunity reactions associated with beta cell destruction. Type 1 DM is a multifactorial disease triggered by genetic and environmental factors, which leads to the destruction of pancreatic beta cells. Early marker of "beta cell autoreactivity" is the synthesis of autoantibodies against 65-kDa protein, which can be a molecule that can be detected early in the disease pathomechanism. The importance of early diagnosis of diabetic patients held in the phase of pre-disease is to determine the progression towards the onset of pancreatic beta cell destruction and take precautions. However, the price for this examination is very expensive ($ 150/ test), the anti-GAD65 abs examination cannot be carried out routinely in most or even in all laboratories in Indonesia. Therefore, production-based Rapid Test Recombinant Human Protein GAD65 with "Reverse Flow Immunchromatography Technique" in Indonesia is believed to reduce costs and improve the quality of care of patients with diabetes in Indonesia. Rapid Test Product innovation is very simple and suitable for screening and routine inspection of GAD65 autoantibodies. In the blood serum of patients with diabetes caused by autoimmunity, autoantibody-GAD65 is a major serologic marker to detect autoimmune reaction because their concentration level of stability.GAD65 autoantibodies can be found 10 years before clinical symptoms of diabetes. Early diagnosis is more focused to detect the presence autontibodi-GAD65 given specification and high sensitivity. Autoantibodies- GAD65 that circulates in the blood is a major indicator of the destruction of the islet cells of the pancreas. Results of research in collaboration with Biofarma has produced GAD65 autoantibodies based Rapid Test had conducted the soft launch of products and has been tested with the results of a sensitivity of 100 percent and a specificity between 90 and 96% compared with the gold standard (import product) which worked based on ELISA method.

Keywords: diabetes mellitus, GAD65 autoantibodies, rapid test, sensitivity, specificity

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Authors: Pao-Huei Chen, Shu-Mei Lin, Yih-Ming Weng, Zer-Ran Yu, Be-Jen Wang

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Pancreatic α-amylase and intestinal α-glucosidase are the critical enzymes for the breakdown of complex carbohydrates into di- or mono-saccharide, which play an important role in modulating postprandial blood sugars. Angiotensin converting enzyme (ACE) converts inactive angiotensin-I into active angiotensin-II, which subsequently increase blood pressure through triggering vasoconstriction and aldosterone secretion. Thus, inhibition of carbohydrate-digestion enzymes and ACE will help the management of blood glucose and blood pressure, respectively. Studies showed Pleurotus citrinopileatus (PC), an edible mushroom and commonly cultured in oriental countries, exerted anticancer, immune improving, antioxidative, hypoglycemic and hypolipidemic effects. Previous studies also showed various molecular weights (MW) fractioned from extracts may affect biological activities due to varying contents of bioactive components. Thus, the objective of this study is to investigate the in vitro antioxidant, hypoglycemic and hypotenstive effects and distribution of active compounds of various MWs of cold water extract from P. citrinopileatus (CWEPC). CWEPC was fractioned into four various MW fractions, PC-I (＜1 kDa), PC-II (1-3.5 kDa), PC-III (3.5-10 kDa), and PC-IV (＞10 kDa), using an ultrafiltration system. The physiological activities, including antioxidant activities, the inhibition capabilities of pancreatic α-amylase, intestinal α-glucosidase, and hypertension-linked ACE, and the active components, including polysaccharides, protein, and phenolic contents, of CWEPC and four fractions were determined. The results showed that fractions with lower MW exerted a higher antioxidant activity (p<0.05), which was positively correlated to the levels of total phenols. In contrast, the inhibition effects on the activities of α-amylase, α-glucosidase, and ACE of PC-IV fraction were significantly higher than CWEPC and the other three low MW fractions (< 10 kDa), which was more related to protein contents. The inhibition capability of CWEPC and PC-IV on α-amylase activity was 1/13.4 to 1/2.7 relative to that of acarbose (positive control), respectively. However, the inhibitory ability of PC-IV on α-glucosidase (IC50 = 0.5 mg/mL) was significantly higher than acarbose (IC50 = 1.7 mg/mL). Kinetic data revealed that PC-IV fraction followed a non-competitive inhibition on α-glucosidase activity. In conclusion, the distribution of various bioactive components contribute to the functions of different MW fractions on oxidative stress prevention, and blood pressure and glucose modulation.

Keywords: α-Amylase, angiotensin converting enzyme, α-Glucosidase, Pleurotus citrinopileatus

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Authors: Oğuz Taşbozan, Celal Erbaş, Şefik Surhan Tabakoğlu, Mahmut Ali Gökçe

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Polyunsaturated fatty acids (PUFAs) and particularly the levels and ratios of ω-3 and ω-6 fatty acids are important for biological functions in humans and recognized as essential components of human diet. According to the terms of many different points of view, the nutritional composition of fish in culture conditions and caught from wild are wondered by the consumers. Therefore the aim of this study was to investigate the chemical composition of cage-cultured and wild sharpsnout seabream which has been preferred by the consumers as an economical important fish species in Turkey. The fish were caught from wild and obtained from cage-cultured commercial companies. Eight fish were obtained for each group, and their average weights of the samples were 245.8±13.5 g for cultured, 149.4±13.3 g for wild samples. All samples were stored in freezer (-18 °C) and analyses were carried out in triplicates, using homogenized boneless fish fillets. Proximate compositions (protein, ash, moisture and lipid) were determined. The fatty acid composition was analyzed by a GC Clarous 500 with auto sampler (Perkin–Elmer, USA). Proximate compositions of cage-cultured and wild samples of sharpsnout seabream were found statistical differences in terms of proximate composition between the groups. The saturated fatty acid (SFA), monounsaturated fatty acid (MUFA) and PUFA amounts of cultured and wild sharpsnout seabream were significantly different. ω3/ω6 ratio was higher in the cultured group. Especially in protein level and lipid level of cultured samples was significantly higher than wild counterparts. One of the reasons for this, cultured species exposed to continuous feeding. This situation had a direct effect on their body lipid content. The fatty acid composition of fish differs depending on a variety of factors including species, diet, environmental factors and whether they are farmed or wild. The higher levels of MUFA in the cultured fish may be explained with the high content of monoenoic fatty acids in the feed of cultured fish as in some other species. The ω3/ω6 ratio is a good index for comparing the relative nutritional value of fish oils. In our study, the cultured sharpsnout seabream appears to be better nutritious in terms of ω3/ω6. Acknowledgement: This work was supported by the Scientific Research Project Unit of the University of Cukurova, Turkey under grant no FBA-2016-5780.

Keywords: Diplodus puntazo, cage cultured, PUFA, fatty acid

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Authors: Mario Hanna Louis Hanna

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The use of CAD (pc Aided layout) generation is ubiquitous inside the structure, engineering and construction (AEC) industry. This has led to its inclusion in the curriculum of structure faculties in Nigeria as an important part of the training module. This newsletter examines the moral troubles involved in implementing CAD (pc Aided layout) content into the architectural training curriculum. Using current literature, this study begins with the advantages of integrating CAD into architectural education and the responsibilities of various stakeholders in the implementation process. It also examines issues related to the terrible use of records generation and the perceived bad effect of CAD use on design creativity. The use of a survey technique, information from the architecture department of Chukwuemeka Odumegwu Ojukwu Uli college changed into accumulated to serve as a case observe on how the problems raised have been being addressed. The object draws conclusions on what guarantees a hit moral implementation. Tens of millions of human beings around the sector suffer from hepatitis C, one of the international's deadliest sicknesses. Interferon (IFN) is a remedy alternative for patients with hepatitis C, but these treatments have their aspect outcomes. Our research targeted growing an oral small molecule drug that goals hepatitis C virus (HCV) proteins and has fewer facet effects. Our contemporary study targets to broaden a drug primarily based on a small molecule antiviral drug precise for the hepatitis C virus (HCV). Drug improvement and the use of laboratory experiments isn't always best high-priced, however also time-eating to behavior those experiments. instead, on this in silicon have a look at, we used computational strategies to recommend a particular antiviral drug for the protein domain names of discovered in the hepatitis C virus. This examines used homology modeling and abs initio modeling to generate the 3-D shape of the proteins, then figuring out pockets within the proteins. Proper lagans for pocket pills were advanced the usage of the de novo drug design method. Pocket geometry is taken into consideration while designing ligands. A few of the various lagans generated, a different for each of the HCV protein domains has been proposed.

Keywords: drug design, anti-viral drug, in-silicon drug design, Hepatitis C virus (HCV) CAD (Computer Aided Design), CAD education, education improvement, small-size contractor automatic pharmacy, PLC, control system, management system, communication.

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Authors: Farahnaz Sadat Golestan Hashemi, Mohd Razi Ismail, Mohd Y. Rafii

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Clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated protein (Cas) system can facilitate targeted genome editing in organisms. Dual or single guide RNA (gRNA) can program the Cas9 nuclease to cut target DNA in particular areas; thus, introducing concise mutations either via error-prone non-homologous end-joining repairing or via incorporating foreign DNAs by homologous recombination between donor DNA and target area. In spite of high demand of such promising technology, developing a well-organized procedure in order for reliable mining of potential target sites for gRNAs in large genomic data is still challenging. Hence, we aimed to perform high-throughput detection of target sites by specific PAMs for not only common Streptococcus pyogenes (SpCas9) but also for Neisseria meningitides (NmCas9) CRISPR-Cas systems. Previous research confirmed the successful application of such RNA-guided Cas9 orthologs for effective gene targeting and subsequently genome manipulation. However, Cas9 orthologs need their particular PAM sequence for DNA cleavage activity. Activity levels are based on the sequence of the protospacer and specific combinations of favorable PAM bases. Therefore, based on the specific length and sequence of PAM followed by a constant length of the target site for the two orthogonals of Cas9 protein, we created a reliable procedure to explore possible gRNA sequences. To mine CRISPR target sites, four different searching modes of sgRNA binding to target DNA strand were applied. These searching modes are as follows i) coding strand searching, ii) anti-coding strand searching, iii) both strand searching, and iv) paired-gRNA searching. Finally, a complete list of all potential gRNAs along with their locations, strands, and PAMs sequence orientation can be provided for both SpCas9 as well as another potential Cas9 ortholog (NmCas9). The artificial design of potential gRNAs in a genome of interest can accelerate functional genomic studies. Consequently, the application of such novel genome editing tool (CRISPR/Cas technology) will enhance by presenting increased versatility and efficiency.

Keywords: CRISPR/Cas9 genome editing, gRNA mining, SpCas9, NmCas9

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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: Agnieszka Krzemińska, Katarzyna Świderek, Vicente Molinier, Piotr Paneth

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In order to understand in details the interactions between ligands and the enzyme isotope effects were studied between clinically used drugs that bind in the active site of Human Immunodeficiency Virus Reverse Transcriptase, HIV-1 RT, as well as triazole-based inhibitor that binds in the allosteric pocket of this enzyme. The magnitudes and origins of the resulting binding isotope effects were analyzed. Subsequently, binding isotope effect of the same triazole-based inhibitor bound in the active site were analyzed and compared. Together, these results show differences in binding origins in two sites of the enzyme and allow to analyze binding mode and place of newly synthesized inhibitors. Typical protocol is described below on the example of triazole ligand in the allosteric pocket. Triazole was docked into allosteric cavity of HIV-1 RT with Glide using extra-precision mode as implemented in Schroedinger software. The structure of HIV-1 RT was obtained from Protein Data Bank as structure of PDB ID 2RKI. The pKa for titratable amino acids was calculated using PROPKA software, and in order to neutralize the system 15 Cl- were added using tLEaP package implemented in AMBERTools ver.1.5. Also N-terminals and C-terminals were build using tLEaP. The system was placed in 144x160x144Å3 orthorhombic box of water molecules using NAMD program. Missing parameters for triazole were obtained at the AM1 level using Antechamber software implemented in AMBERTools. The energy minimizations were carried out by means of a conjugate gradient algorithm using NAMD. Then system was heated from 0 to 300 K with temperature increment 0.001 K. Subsequently 2 ns Langevin−Verlet (NVT) MM MD simulation with AMBER force field implemented in NAMD was carried out. Periodic Boundary Conditions and cut-offs for the nonbonding interactions, range radius from 14.5 to 16 Å, are used. After 2 ns relaxation 200 ps of QM/MM MD at 300 K were simulated. The triazole was treated quantum mechanically at the AM1 level, protein was described using AMBER and water molecules were described using TIP3P, as implemented in fDynamo library. Molecules 20 Å apart from the triazole were kept frozen, with cut-offs established on range radius from 14.5 to 16 Å. In order to describe interactions between triazole and RT free energy of binding using Free Energy Perturbation method was done. The change in frequencies from ligand in solution to ligand bounded in enzyme was used to calculate binding isotope effects.

Keywords: binding isotope effects, molecular dynamics, HIV, reverse transcriptase

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Authors: Raka Rahmatulloh, Mohammad Ilham Nugraha, Muhammad Ifan Fathurrahman

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The agricultural sector covers a wide area, one of them is livestock subsector that supply needs of the food source of animal protein. Animal protein is produced by the main livestock production such as meat, milk, eggs, etc. Besides the main production, livestock would produce metabolic residue, so called livestock wastes. Characteristics of livestock wastes can be either solid (feces), liquid (urine), and gas (methane) which turned out to be useful and has economical value when well-processed and well-controlled. Nowadays, this livestock wastes is considered as a source of pollutants, especially water pollution. If the source of pollutants used in an integrated way, it will have a positive impact on organic farming and a healthy environment. Management of livestock wastes can be integrated with the farming sector to the planting and caring that rely on fertilizers. Most Indonesian farmers still use chemical fertilizers, where the use of it in the long term will disturb the ecological balance of the environment. One of the main efforts is to use organic fertilizers instead of chemical fertilizer that conducted by the Faculty of Animal Husbandry, Padjadjaran University. The method is to use the solid waste of livestock and agricultural wastes into liquid organic fertilizer, feed additive, biogas and vermicompost through decomposition. The decomposition takes as long as 14 days including aeration and extraction process using water as a nutrients solvent media which contained in decomposes and disinfection media to release pathogenic microorganisms in decomposes. Liquid organic fertilizer has highly efficient for the farmers to have a ratio of carbon/nitrogen (C/N) 25/1 to 30/1 and neutral pH (6.5-7.5) which is good for plant growth. Feed additive may be given to improve the digestibility of feed so that substances can be easily absorbed by the body for production. Biogas contains methane (CH4), which has a high enough heat to produce electricity. Vermicompost is an overhaul of waste organic material that has excellent structure, porosity, aeration, drainage, and moisture holding capacity. Based on the case study above, an integrated livestock wastes management program strongly supports the Indonesian government in the achievement of sustainable livestock development.

Keywords: integrated, livestock wastes, organic fertilizer, sustainable livestock development

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Authors: Marta Fernández Batalla, José María Santamaría García, Maria Lourdes Jiménez Rodríguez, Roberto Barchino Plata, Adriana Cercas Duque, Enrique Monsalvo San Macario

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Background: Primary Care Nursing is taking more autonomy in clinical decisions. One of the most frequent therapies to solve is related to the problems of maintaining a sufficient supply of food. Nursing diagnoses related to food are addressed by the nurse-family and community as the first responsible. Objectives and interventions are set according to each patient. To improve the goal setting and the treatment of these care problems, a technological tool is developed to help nurses. Objective: To evaluate the computational tool developed to support the clinical decision in feeding problems. Material and methods: A cross-sectional descriptive study was carried out at the Meco Health Center, Madrid, Spain. The study population consisted of four specialist nurses in primary care. These nurses tested the tool on 30 people with ‘need for nutritional therapy’. Subsequently, the usability of the tool and the satisfaction of the professional were sought. Results: A simple and convenient computational tool is designed for use. It has 3 main entrance fields: age, size, sex. The tool returns the following information: BMI (Body Mass Index) and caloric consumed by the person. The next step is the caloric calculation depending on the activity. It is possible to propose a goal of BMI or weight to achieve. With this, the amount of calories to be consumed is proposed. After using the tool, it was determined that the tool calculated the BMI and calories correctly (in 100% of clinical cases). satisfaction on nutritional assessment was ‘satisfactory’ or ‘very satisfactory’, linked to the speed of operations. As a point of improvement, the options of ‘stress factor’ linked to weekly physical activity. Conclusion: Based on the results, it is clear that the computational tools of decision support are useful in the clinic. Nurses are not only consumers of computational tools, but can develop their own tools. These technological solutions improve the effectiveness of nutrition assessment and intervention. We are currently working on improvements such as the calculation of protein percentages as a function of protein percentages as a function of stress parameters.

Keywords: feeding behavior health, nutrition therapy, primary care nursing, technology assessment

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Authors: Jean De Dieu Ayabagabo, Paul A.Onjoro, Karubiu P. Migwi, Marie C. Dusingize

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This study investigated the effects of feed on milk yield and quality through feed monitoring and quality assessment, and the consequent enteric methane gas emissions from smallholder dairy farms in drier areas of Rwanda, using the Tier II approach for four seasons in three zones, namely; Mayaga and peripheral Bugesera (MPB), Eastern Savanna and Central Bugesera (ESCB), and Eastern plateau (EP). The study was carried out using 186 dairy cows with a mean live weight of 292 Kg in three communal cowsheds. The milk quality analysis was carried out on 418 samples. Methane emission was estimated using prediction equations. Data collected were subjected to ANOVA. The dry matter intake was lower (p<0.05) in the long dry season (7.24 Kg), with the ESCB zone having the highest value of 9.10 Kg, explained by the practice of crop-livestock integration agriculture in that zone. The Dry matter digestibility varied between seasons and zones, ranging from 52.5 to 56.4% for seasons and from 51.9 to 57.5% for zones. The daily protein supply was higher (p<0.05) in the long rain season with 969 g. The mean daily milk production of lactating cows was 5.6 L with a lower value (p<0.05) during the long dry season (4.76 L), and the MPB zone having the lowest value of 4.65 L. The yearly milk production per cow was 1179 L. The milk fat varied from 3.79 to 5.49% with a seasonal and zone variation. No variation was observed with milk protein. The seasonal daily methane emission varied from 150 g for the long dry season to 174 g for the long rain season (p<0.05). The rain season had the highest methane emission as it is associated with high forage intake. The mean emission factor was 59.4 Kg of methane/year. The present EFs were higher than the default IPPC value of 41 Kg from developing countries in African, the Middle East, and other tropical regions livestock EFs using Tier I approach due to the higher live weight in the current study. The methane emission per unit of milk production was lower in the EP zone (46.8 g/L) due to the feed efficiency observed in that zone. Farmers should use high-quality feeds to increase the milk yield and reduce the methane gas produced per unit of milk. For an accurate assessment of the methane produced from dairy farms, there is a need for the use of the Life Cycle Assessment approach that considers all the sources of emissions.

Keywords: footprint, forage, girinka, tier

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Authors: Laura Rueda-Gensini, Jader Rodríguez, Juan C. Cruz, Carolina Munoz-Camargo

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There is an emerging interest in botanicals and plant extracts for medicinal practices due to their widely reported health benefits. A large variety of phytochemicals found in plants have been correlated with antioxidant, immunomodulatory, and analgesic properties, which makes plant-derived products promising candidates for modulating the progression and treatment of numerous diseases. Non-centrifugal cane sugar (NCS), in particular, has been known for its high antioxidant and nutritional value, but composition-wise variability due to changing environmental and processing conditions have considerably limited its use in the nutraceutical and biomedical fields. This work is therefore aimed at assessing the effect of thermal exposure during NCS production over its bioactive composition and, in turn, its therapeutic value. Accordingly, two modified dehydration methods are proposed that employ: (i) vacuum-aided evaporation, which reduces the necessary temperatures to dehydrate the sample, and (ii) window refractance evaporation, which reduces thermal exposure time. The biochemical composition of NCS produced under these two methods was compared to traditionally-produced NCS by estimating their total polyphenolic and protein content with Folin-Ciocalteu and Bradford assays, as well as identifying the major phenolic compounds in each sample via HPLC-coupled mass spectrometry. Their antioxidant activities were also compared as measured by their scavenging potential of ABTS and DPPH radicals. Results show that the two modified production methods enhance polyphenolic and protein yield in resulting NCS samples when compared to traditional production methods. In particular, reducing employed temperatures with vacuum-aided evaporation demonstrated to be superior at preserving polyphenolic compounds, as evidenced both in the total and individual polyphenol concentrations. However, antioxidant activities were not significantly different between these. Although additional studies should be performed to determine if the observed compositional differences affect other therapeutic activities (e.g., anti-inflammatory, analgesic, and immunoprotective), these results suggest that reducing thermal exposure holds great promise for the production of natural products with enhanced nutritional value.

Keywords: non-centrifugal cane sugar, polyphenolic compounds, thermal processing, antioxidant activity

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Authors: Bekmurza H. Aitchanov, Sholpan K. Aitchanova, Olimzhon A. Baimuratov, Aitkul N. Aldibekova

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This work considers the automated control system (ACS) of milk quality during its magnetic field processing. For achieving high level of quality control methods were applied transformation of complex nonlinear systems in a linearized system with a less complex structure. Presented ACS is adjustable by seven parameters: mass fraction of fat, mass fraction of dry skim milk residues (DSMR), density, mass fraction of added water, temperature, mass fraction of protein, acidity.

Keywords: fluids magnetization, nuclear magnetic resonance, automated control system, dynamic pulse-frequency modulator, PFM, nonlinear systems, structural model

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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: Nseabasi Nsikakabasi Etim

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In recent times, there has been a significant shortfall in between the production and supply of animal protein to meet the ever increasing population. To meet the increasing demand for animal protein, there is a need to focus attention on the production of livestock whose nutritional requirement does not put much strain on the limited sources of feed ingredients to which men subscribe. An example of such livestock is the rabbit. Rabbit is a pseudo-ruminant herbivore which utilizes much undigested and unabsorbed feed materials as sources of nutrient for maintenance and production. Thus, this study was conducted to investigate the effects of feeding Aspilia africana as forage on the growth rates of female pseudo-ruminant herbivores (rabbits) at different physiological states. Thirty (30) Dutch breed rabbit does of 5–6 months of age were used for the experiment which was conducted in a completely randomized design for four months. The rabbits were divided into three treatment groups, ten does per treatment group; which consisted of mixed forages (Centrosema pubescent (200g), Panicum maximum (200g) and Ipomea batatas leaves (100g) without Aspilia africana (T1; control), fresh Aspilia africana (500g/dose/day) (T2) and wilted Aspilia africana (500g/dose/day) (T3). Rabbits in all treatment groups received the same concentrate (300g/animal/day) throughout the period of the study and mixed forages from the commencement of the experiment till the does kindled. After parturition, fresh and wilted Aspilia africana were introduced in treatments 2 and three respectively, whereas the control group continued on mixed forages throughout the study. The result of the study revealed that the initial average body weight of the rabbit does was 1.74kg. At mating and gestation periods, the body weights of the does in T2 was significantly higher (P<0.05) than the rest. There were no significant differences (P<0.05) in the body weights of does at kindling between the various treatment groups. During the physiological states of lactation, weaning and re-mating, the control group (T1) had significantly lower body weight than those of the treated groups (T2 and T3). Furthermore, T2 had significantly higher body weight than T3. The study revealed that Aspilia africana; mainly the fresh leaves have greater growth stimulating effects when fed to pseudo-ruminants (rabbits), thereby enhancing body weights of does during lactation and weaning.

Keywords: Aspilia africana, herbivores, pseudo-ruminants, physiological states

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Authors: Turban Kar, Maitree Bhattacharyya

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Curcumin, a dietary antioxidant has been identified as a wonder molecule to possess therapeutic properties protecting the cellular macromolecules from oxidative damage. In our experimental study, we have explored the effectiveness of curcumin in protecting the structural paradigm of Human Serum Albumin (HSA) when exposed to gamma irradiation. HSA, being an important transport protein of the circulatory system, is involved in binding of variety of metabolites, drugs, dyes and fatty acids due to the presence of hydrophobic pockets inside the structure. HSA is also actively involved in the transportation of drugs and metabolites to their targets, because of its long half-life and regulation of osmotic blood pressure. Gamma rays, in its increasing concentration, results in structural alteration of the protein and superoxide radical generation. Curcumin, on the other hand, mitigates the damage, which has been evidenced in the following experiments. Our study explores the possibility for protection by curcumin during the molecular and conformational changes of HSA when exposed to gamma irradiation. We used a combination of spectroscopic methods to probe the conformational ensemble of the irradiated HSA and finally evaluated the extent of restoration by curcumin. SDS - PAGE indicated the formation of cross linked aggregates as a consequence of increasing exposure of gamma radiation. CD and FTIR spectroscopy inferred significant decrease in alpha helix content of HSA from 57% to 15% with increasing radiation doses. Steady state and time resolved fluorescence studies complemented the spectroscopic measurements when lifetime decay was significantly reduced from 6.35 ns to 0.37 ns. Hydrophobic and bityrosine study showed the effectiveness of curcumin for protection against radiation induced free radical generation. Moreover, bityrosine and hydrophobic profiling of gamma irradiated HSA in presence and absence of curcumin provided light on the formation of ROS species generation and the protective (magical) role of curcumin. The molecular mechanism of curcumin protection to HSA from gamma irradiation is yet unknown, though a possible explanation has been proposed in this work using Thioflavin T assay. It was elucidated, that when HSA is irradiated at low dose of gamma radiation in presence of curcumin, it is capable of retaining the native characteristic properties to a greater extent indicating stabilization of molecular structure. Thus, curcumin may be utilized as a therapeutic strategy to protect cellular proteins.

Keywords: Bityrosine content, conformational change, curcumin, gamma radiation, human serum albumin

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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: K. Thiel, P. Patrikainen, C. Nagy, D. Fitzpatrick, E.-M. Aro, P. Kallio

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Photosynthetic cyanobacteria have been recognized as potential future biotechnological hosts for the direct conversion of CO₂ into chemicals of interest using sunlight as the solar energy source. However, in order to develop commercially viable systems, the flux of electrons from the photosynthetic light reactions towards specified target chemicals must be significantly improved. The objective of the study was to investigate whether the autotrophic production efficiency of specified end-metabolites can be improved in engineered cyanobacterial cells by rescuing excited electrons that are normally lost to molecular oxygen due to the cyanobacterial flavodiiron protein Flv1/3. Natively Flv1/3 dissipates excess electrons in the photosynthetic electron transfer chain by directing them to molecular oxygen in Mehler-like reaction to protect photosystem I. To evaluate the effect of flavodiiron inactivation on autotrophic production efficiency in the cyanobacterial host Synechocystis sp. PCC 6803 (Synechocystis), sucrose was selected as the quantitative reporter and a representative of a potential end-product of interest. The concept is based on the native property of Synechocystis to produce sucrose as an intracellular osmoprotectant when exposed to high external ion concentrations, in combination with the introduction of a heterologous sucrose permease (CscB from Escherichia coli), which transports the sucrose out from the cell. In addition, cell growth, photosynthetic gas fluxes using membrane inlet mass spectrometry and endogenous storage compounds were analysed to illustrate the consequent effects of flv deletion on pathway flux distributions. The results indicate that a significant proportion of the electrons can be lost to molecular oxygen via Flv1/3 even when the cells are grown under high CO₂ and that the inactivation of flavodiiron activity can enhance the photosynthetic electron flux towards optionally available sinks. The flux distribution is dependent on the light conditions and the genetic context of the Δflv mutants, and favors the production of either sucrose or one of the two storage compounds, glycogen or polyhydroxybutyrate. As a conclusion, elimination of the native Flv1/3 reaction and concomitant introduction of an engineered product pathway as an alternative sink for excited electrons could enhance the photosynthetic electron flux towards the target endproduct without compromising the fitness of the host.

Keywords: cyanobacterial engineering, flavodiiron proteins, redirecting electron flux, sucrose

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Authors: Elvan G. Bulut, Betul Goksun, Tugba G. Gun, Ozge Sakiyan Demirkol, Kamuran Ayhan, Kezban Candogan

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Dysphagia, difficulty in swallowing solid foods and thin liquids, is one of the common health threats among the elderly who require foods with modified texture in their diet. Although there are some commercial food formulations or hydrocolloids to thicken the liquid foods for dysphagic individuals, there is still a need for developing and offering new food products with enriched nutritional, textural and sensory characteristics to safely nourish these patients. 3D food printing is an appealing alternative in creating personalized foods for this purpose with attractive shape, soft and homogenous texture. In order to modify texture and prevent phase separation, hydrocolloids are generally used. In our laboratory, an optimized 3D printed beef-based formulation specifically for people with swallowing difficulties was developed based on the research project supported by the Scientific and Technological Research Council of Turkey (TÜBİTAK Project # 218O017). The optimized formulation obtained from response surface methodology was 60% beef powder, 5.88% gelatin, and 0.74% kappa-carrageenan (all in a dry basis). This product was enriched with powders of freeze-dried beet, celery, and red capia pepper, butter, and whole milk. Proximate composition (moisture, fat, protein, and ash contents), pH value, CIE lightness (L*), redness (a*) and yellowness (b*), and color difference (ΔE*) values were determined. Counts of total mesophilic aerobic bacteria (TMAB), lactic acid bacteria (LAB), mold and yeast, total coliforms were conducted, and detection of coagulase positive S. aureus, E. coli, and Salmonella spp. were performed. The 3D printed products had 60.11% moisture, 16.51% fat, 13.68% protein, and 1.65% ash, and the pH value was 6.19, whereas the ΔE* value was 3.04. Counts of TMAB, LAB, mold and yeast and total coliforms before and after 3D printing were 5.23-5.41 log cfu/g, < 1 log cfu/g, < 1 log cfu/g, 2.39-2.15 log EMS/g, respectively. Coagulase positive S. aureus, E. coli, and Salmonella spp. were not detected in the products. The data obtained from this study based on determining some important product characteristics of functional beef-based formulation provides an encouraging basis for future research on the subject and should be useful in designing mass production of 3D printed products of similar composition.

Keywords: beef, dysphagia, product characteristics, texture-modified foods, 3D food printing

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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: Alice Nwanyioma Ohuoba, Rose Erdoo Kukwa, Ukpabi Joseph Ukpabi

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Cassava root (Manihot esculenta) is one of the important carbohydrates containing crops in Nigeria. It is a staple food, mostly in the southern part of the country, and a source of income to farmers and processors. Cassava gari processing methods result to residue fiber (solid waste) from the sieving operation, these residue fibers ( solid wastes) can be dried and milled into flour and used to prepare cakes, cookies, crackers and chin-chin instead of being thrown away mostly on farmland or near the residential area. Flour for baking or frying may contain carbohydrates and protein (wheat flour) or rich in only carbohydrates (cassava flour). Cake, cookies, crackers, and chin-chin were prepared using the residue flour obtained from the residue fiber of cassava variety NR87184 roots, processed into gari. This study is aimed at evaluating the proximate composition, mineral content and sensory attributes of these selected snacks produced. The proximate composition results obtained showed that crackers had the lowest value in moisture (2.3390%) and fat (1.7130%), but highest in carbohydrates (85.2310%). Amongst the food products, cakes recorded the highest value in protein (8.0910%). Crude fibre values ranges from 2.5265% (cookies) to 3.4165% (crackers). The result of the mineral contents showed cookies ranking the highest in Phosphorus (65.8535 ppm) and Iron (0.1150 mg/L), Calcium (1.3800mg/L) and Potassium (7.2850 mg/L) contents, while chin-chin and crackers were lowest in Sodium ( 2.7000 mg/L). The food products were also subjected to sensory attributes evaluation by thirty member panelists using 9-hedonic scale which ranged from 1 ( dislike extremely) to 9 (like extremely). The means score obtained shows all the food products having above 7.00 (above “like moderately”). This study has shown that food products that may be functional or nutraceuticals could be prepared from the residue flour. There is a call for the use of gluten-free flour in baking due to ciliac disease and other allergic causes by gluten. Therefore local carbohydrates food crops like cassava residue flour that are gluten-free, could be the solution. In addition, this could aid cassava gari processing waste management thereby reducing post-harvest losses of cassava root.

Keywords: allergy, flour, food-products, gluten-free

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Authors: Valencia Fernandes, Dharmendra Kumar Khatri, Shashi Bala Singh

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Background and Aim: Epigenetics are the inaudible signatures of several pathological processes in the brain. This study understands the influence of DNA methylation, a major epigenetic modification, in the prefrontal cortex and hippocampus of the diabetic brain and its notable effect on the cellular chaperones and synaptic proteins. Method: Chronic high fat diet and STZ-induced diabetic mice were studied for cognitive dysfunction, and global DNA methylation, as well as DNA methyltransferase (DNMT) activity, were assessed. Further, the cellular chaperones and synaptic proteins were examined using DNMT inhibitor, 5-aza-2′-deoxycytidine (5-aza-dC)-via intracerebroventricular injection. Moreover, % methylation of these synaptic proteins were also studied so as to correlate its epigenetic involvement. Computationally, its interaction with the DNMT enzyme were also studied using bioinformatic tools. Histological studies for morphological alterations and neuronal degeneration were also studied. Neurogenesis, a characteristic marker for new learning and memory formation, was also assessed via the BrdU staining. Finally, the most important behavioral studies, including the Morris water maze, Y maze, passive avoidance, and Novel object recognition test, were performed to study its cognitive functions. Results: Altered global DNA methylation and increased levels of DNMTs within the nucleus were confirmed in the cortex and hippocampus of the diseased mice, suggesting hypermethylation at a genetic level. Treatment with AzadC, a global DNA demethylating agent, ameliorated the protein and gene expression of the cellular chaperones and synaptic fidelity. Furthermore, the methylation analysis profile showed hypermethylation of the hsf1 protein, a master regulator for chaperones and thus, confirmed the epigenetic involvement in the diseased brain. Morphological improvements and decreased neurodegeneration, along with enhanced neurogenesis in the treatment group, suggest that epigenetic modulations do participate in learning and memory. This is supported by the improved behavioral test battery seen in the treatment group. Conclusion: DNA methylation could possibly accord in dysregulating the memory-associated proteins at chronic stages in type 2 diabetes. This could suggest a substantial contribution to the underlying pathophysiology of several metabolic syndromes like insulin resistance, obesity and also participate in transitioning this damage centrally, such as cognitive dysfunction.

Keywords: epigenetics, cognition, chaperones, DNA methylation

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