Search results for: candidate gene of type-2 diabetes
Commenced in January 2007
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Edition: International
Paper Count: 2902

Search results for: candidate gene of type-2 diabetes

2602 Phytochemicals from Enantia Chlorantha Stem Bark Inhibits the Activity ?-Amylase and ?-Glucosidase: Molecular Docking Studies

Authors: Hammed Tanimowo Aiyelabegan, Oluchukwu Franklin Aladi, Mutiu Adewumi Alabi, Raliat Abimbola Aladodo, Emmanuel Oladipupo Ajani, Abdulganiyu Giwa, Esther Owolabi

Abstract:

The study aimed to evaluate the inhibitory activities of ligands from Enantia chlorantha stem bark on α-amylase and α-glucosidase. In silico pharmacokinetic properties and docking scores were employed to analyse the inhibition using SwissADME and Autodock4.2, respectively. Results revealed that drug-likeness, pharmacokinetics and bioavailability radar of all the ligands except jatrorrhizine and acarbose falls within the radar according to the Lipinski rule of 5. The binding energies of the protein-ligand interactions also show that the ligand fits into the active site. The results obtained from this study show that the chemical constituents from Enantia chlorantha stem bark may bring about positive physiological changes in a patient suffering from diabetes mellitus. Further in vitro studies on diabetes cell lines and in vivo studies on the animal may validate these compounds for diabetes treatment. These phytoconstituents could help in the development of novel anti-diabetic molecules.

Keywords: diabetes mellitus, ?-amylase, ?-glucosidase, in silico, Enantia chlorantha stem bark

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2601 Co-Existence of Central Serous Retinopathy and Diabetic Retinopathy: A Diagnostic Dilemma

Authors: Avantika Verma

Abstract:

Diabetic retinopathy (DR) and Central serous retinopathy (CSR) are 2 distinct entities, with difference in age of presentation, eitiopathogenesis and clinical features, but when occurring together, can be a diagnostic dilemma and requires careful evaluation. Case study of 3 patients with long standing diabetes (>15yrs) and features of Central serous retinopathy was done at Bangalore West Lions Superspeciality Eye Hospital, Bangalore, India in 2013. Even though diabetic retinopathy and CSR have different pathologies, they can coexist. The reason for coexistence could be the following: A patient with CSR as a young adult could develop DR in later years. Stress could be the contributing factor in older patient with diabetes.Stress could be a common factor for both, as it is one of the important factors in the pathogenesis of Maturity Onset Diabetes Miletus (MODY). In any situation, a careful evaluation is necessary to differentiate the cause of fundus picture, as treatment differs for the two diseases.

Keywords: central serous retinopathy, diabetic retinopathy, existence, stress

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2600 Variation in Carboxylesterase Activity in Spodoptera litura Fabricious (Noctuidae: Lepidoptera) Populations from India

Authors: V. Karuppaiah, J. C. Padaria, C. Srivastava

Abstract:

The tobacco caterpillar, Spodoptera litura Fab (Lepidoptera: Noctuidae) is a polyphagous pest various field and horticulture crops in India. Pest had virtually developed resistance to all commonly used insecticides. Enhanced detoxification is the prime mechanism that is dictated by detoxification different enzymes and carboxylesterase is one of the major enzyme responsible development of resistance. In India, insecticide resistance studies on S. litura are mainly deployed on detoxification enzymes activity and investigation at gene level alteration i.e. at nucleotide level is very merger. In the present study, we collected the S. litura larvae from three different cauliflower growing belt viz., IARI, New Delhi (Delhi), Palari, Sonepat (Haryana) and Varanasi (Uttar Pradesh) to study the role of carboxylesterase activity and its gene level variation The CarE activity was measured using UV-VIS spectrophotometer with 3rd instar larvae of S. litura. The elevated activity of CarE was observed in Sonepat strain (28.09 ± 0.09 µmol/min/mg of protein) followed by Delhi (26.72 ± 0.04 µmol/min/mg of protein) and Varanasi strain (10.00 ± 0.44 µmol/min/mg of protein) of S. litura. The genomic DNA was isolated from 3rd instar larvae and CarE gene was amplified using a primer sequence, F:5’tccagagttccttgtcaggcac3’; R:5’ctgcatcaagcatgtctc3. CarE gene, about 500bp was partially amplified, sequenced and submitted to NCBI (Accession No. KF835886, KF835887 and KF835888). The sequence data revealed polymorphism at nucleotide level in all the three strains and gene found to have 88 to 97% similarity with previous available nucleotide sequences of S. litura, S. littoralis and S. exiqua. The polymorphism at the nucleotide level could be a reason for differential activity of carboxylesterase enzymes among the strains. However, investigation at gene expression level would be useful to analyze the overproduction of carboxylesterase enzyme.

Keywords: carboxylesterase, CarE gene, nucleotide polymorphism, insecticide resistance, spodoptera litura

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2599 Diabetes Diagnosis Model Using Rough Set and K- Nearest Neighbor Classifier

Authors: Usiobaifo Agharese Rosemary, Osaseri Roseline Oghogho

Abstract:

Diabetes is a complex group of disease with a variety of causes; it is a disorder of the body metabolism in the digestion of carbohydrates food. The application of machine learning in the field of medical diagnosis has been the focus of many researchers and the use of recognition and classification model as a decision support tools has help the medical expert in diagnosis of diseases. Considering the large volume of medical data which require special techniques, experience, and high diagnostic skill in the diagnosis of diseases, the application of an artificial intelligent system to assist medical personnel in order to enhance their efficiency and accuracy in diagnosis will be an invaluable tool. In this study will propose a diabetes diagnosis model using rough set and K-nearest Neighbor classifier algorithm. The system consists of two modules: the feature extraction module and predictor module, rough data set is used to preprocess the attributes while K-nearest neighbor classifier is used to classify the given data. The dataset used for this model was taken for University of Benin Teaching Hospital (UBTH) database. Half of the data was used in the training while the other half was used in testing the system. The proposed model was able to achieve over 80% accuracy.

Keywords: classifier algorithm, diabetes, diagnostic model, machine learning

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2598 Gene Distribution of CB1 Receptor rs2023239 in Thailand Cannabis Patients

Authors: Tanyaporn Chairoch

Abstract:

Introduction: Cannabis is a drug to treat patients with many diseases such as Multiple sclerosis, Alzheimer’s disease, and Epilepsy, where theycontain many active compounds such as delta-9 tetrahydrocannabinol (THC) and cannabidiol (CBD). Especially, THC is the primary psychoactive ingredient in cannabis and binds to cannabinoid 1 (CB1) receptors. Moreover, CB1 is located on the neocortex, hippocampus, basal ganglia, cerebellum, and brainstem. In previous study, we found the association between the variant of CB1recptors gene (rs2023239) and decreased effect of nicotine reinforcement in patients. However, there are no data describing whether the distribution of CB1 receptor gene is a genetic marker for Thai patients who are treated with cannabis. Objective: Thus, the aim of this study we want to investigate the frequency of the CB1 receptor gene in Thai patients. Materials and Methods: All of sixty Thai patients received the medical cannabis for treatment who were recruited in this study. DNA will be extracted from EDTA whole blood by Genomic DNA Mini Kit. The genotyping of CNR1 gene (rs 2023239) was genotyped by the TaqMan real time PCR assay (ABI, Foster City, CA, USA).and using the real-time PCR ViiA7 (ABI, Foster City, CA, USA). Results: We found thirty-eight (63.3%) Thai patients were female, and twenty-two (36.70%) were male in this study with median age of 45.8 (range19 – 87 ) years. Especially, thirty-two (53.30%) medical cannabis tolerant controls were female ( 55%) and median age of52.1 (range 27 – 79 ) years. The most adverse effects for medical cannabis treatment was tachycardia. Furthermore, the number of rs 2023239 (TT) carriers was 26 of 27 (96.29%) in medical cannabis-induced adverse effects and 32 of 33 (96.96%) in tolerant controls. Additionally, rs 2023239 (CT) variant was found just only one of twenty-seven (3.7%) in medical cannabis-induced adverse effects and 1 of 33 (3.03%) in tolerant controls. Conclusions: The distribution of genetic variant in CNR1 gene might serve as a pharmacogenetics markers for screening before initiating the therapy with medical cannabis in Thai patients.

Keywords: cannabis, pharmacogenetics, CNR1 gene, thai patient

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2597 Prevalence of Microalbuminuria and Its Relation with Various Risk Factors in Type 1 Diabetes Mellitus

Authors: Singh Baljinder, Sharma Navneet

Abstract:

Microalbuminuria is the earliest detectable marker of diabetic nephropathy. We planned to evaluate the prevalence of microalbuminuria in type 1 diabetics and correlate with various risk factor. We randomly selected 100 type 1 diabetic patients after inclusion and exclusion criteria from DCRC, S. P. Medical College, Bikaner. Clinical examinations for anthropometeric parameters, hypertension, retinopathy, glycaemic status, lipid profile were done and microalbuminuria was estimated by micral test. Microalbuminuria was seen in 38% patients. The mean urinary albumin concentration was 96.61 mg/l in microalbuminuria positive cases, 134 mg/L in hypertensive patients while 74.5 mg/L in normal patients. Mean diabetic duration was 6.43 years in microalbuminurics. Albumin excretion increased significantly with age at onset of 10-18 years and declined thereafter. Microalbuminuria cases exhibited mean cholesterol 181.63 mg%, TG 130.94 mg%, LDL 109.87 mg%, HDL 57.5 mg% and VLDL 30.64 mg%. Mean urinary albumin concentration in patients with retinopathy was 160.52 mg/L while 78.66 mg/L without retinopathy. In multiple stepwise logistic regression analysis, a strong positive association was seen between microalbuminuria and hypertension (OR=5.087, CI=2.1319-12.101), fasting blood sugar (OR=3. 491, CI=1.138-10.70), duration of diabetes (OR=3.41, CI=1.360-8.55) and HbA1c (OR=2.381, CI-=1.1-5.64). The present study indicates that microalbuminuria is a common complication of type 1 diabetes mellitus and can be prevented by careful management of risk factors.

Keywords: type 1 diabetes, microalbuminuria, diabetic nephropathy, retinopathy, hypertension

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2596 Modeling of CREB Pathway Induced Gene Induction: From Stimulation to Repression

Authors: K. Julia Rose Mary, Victor Arokia Doss

Abstract:

Electrical and chemical stimulations up-regulate phosphorylaion of CREB, a transcriptional factor that induces its target gene production for memory consolidation and Late Long-Term Potentiation (L-LTP) in CA1 region of the hippocampus. L-LTP requires complex interactions among second-messenger signaling cascade molecules such as cAMP, CAMKII, CAMKIV, MAPK, RSK, PKA, all of which converge to phosphorylate CREB which along with CBP induces the transcription of target genes involved in memory consolidation. A differential equation based model for L-LTP representing stimulus-mediated activation of downstream mediators which confirms the steep, supralinear stimulus-response effects of activation and inhibition was used. The same was extended to accommodate the inhibitory effect of the Inducible cAMP Early Repressor (ICER). ICER is the natural inducible CREB antagonist represses CRE-Mediated gene transcription involved in long-term plasticity for learning and memory. After verifying the sensitivity and robustness of the model, we had simulated it with various empirical levels of repressor concentration to analyse their effect on the gene induction. The model appears to predict the regulatory dynamics of repression on the L-LTP and agrees with the experimental values. The flux data obtained in the simulations demonstrate various aspects of equilibrium between the gene induction and repression.

Keywords: CREB, L-LTP, mathematical modeling, simulation

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2595 Bean in Turkey: Characterization, Inter Gene Pool Hybridization Events, Breeding, Utilizations

Authors: Faheem Shahzad Baloch, Muhammad Azhar Nadeem, Muhammad Amjad Nawaz, Ephrem Habyarimana, Gonul Comertpay, Tolga Karakoy, Rustu Hatipoglu, Mehmet Zahit Yeken, Vahdettin Ciftci

Abstract:

Turkey is considered a bridge between Europe, Asia, and Africa and possibly played an important role in the distribution of many crops including common bean. Hundreds of common bean landraces can be found in Turkey, particularly in farmers’ fields, and they consistently contribute to the overall production. To investigate the existing genetic diversity and hybridization events between the Andean and Mesoamerican gene pools in the Turkish common bean, 188 common bean accessions (182 landraces and 6 modern cultivars as controls) were collected from 19 different Turkish geographic regions. These accessions were characterized using phenotypic data (growth habit and seed weight), geographic provenance, 12557 high-quality whole-genome DArTseq markers, and 3767 novel DArTseq loci were also identified. The clustering algorithms resolved the Turkish common bean landrace germplasm into the two recognized gene pools, the Mesoamerican and Andean gene pools. Hybridization events were observed in both gene pools (14.36% of the accessions) but mostly in the Mesoamerican (7.97% of the accessions), and was low relative to previous European studies. The lower level of hybridization witnessed the existence of Turkish common bean germplasm in its original form as compared to Europe. Mesoamerican gene pool reflected a higher level of diversity, while the Andean gene pool was predominant (56.91% of the accessions), but genetically less diverse and phenotypically more pure, reflecting farmers greater preference for the Andean gene pool. We also found some genetically distinct landraces and overall, a meaningful level of genetic variability which can be used by the scientific community in breeding efforts to develop superior common bean strains.

Keywords: bean germplasm, DArTseq markers, genotyping by sequencing, Turkey, whole genome diversity

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2594 Socioeconomic Disparities in the Prevalence of Obesity in Adults with Diabetes in Israel

Authors: Yael Wolff Sagy, Yiska Loewenberg Weisband, Vered Kaufman Shriqui, Michal Krieger, Arie Ben Yehuda, Ronit Calderon Margalit

Abstract:

Background: Obesity is both a risk factor and common comorbidity of diabetes. Obesity impedes the achievement of glycemic control, and enhances damage caused by hyperglycemia to blood vessels; thus it increases diabetes-related complications. This study assessed the prevalence of obesity and morbid obesity among Israeli adults with diabetes, and estimated disparities associated with sex and socioeconomic position (SEP). Methods: A cross-sectional study was conducted in the setting of the Israeli National Program for Quality Indicators in Community Healthcare. Data on all the Israeli population is retrieved from electronic medical records of the four health maintenance organizations (HMOs). The study population included all Israeli patients with diabetes aged 20-64 with documented body mass index (BMI) in 2016 (N=180,451). Diabetes was defined as the existence of one or more of the following criteria: (a) Plasma glucose level >200 mg% in at least two tests conducted at least one month apart in the previous year; (b) HbA1c>6.5% at least once in the previous year (c) at least three prescriptions of diabetes medications were dispensed during the previous year. Two measures were included: the prevalence of obesity (defined as last BMI≥ 30 kg/m2 and <35 kg/m2) and the prevalence of morbid obesity (defined as last BMI≥ 35 kg/m2) in individuals aged 20-64 with diabetes. The cut-off value for morbid obesity was set in accordance with the eligibility criteria for bariatric surgery in diabetics. Data were collected by the HMOs and aggregated by age, sex and SEP. SEP was based on statistical areas ranking by the Israeli Central Bureau of Statistics and divided into 4 categories, ranking from 1 (lowest) to 4 (highest). Results: BMI documentation among adults with diabetes was 84.9% in 2016. The prevalence of obesity in the study population was 30.5%. Although the overall rate was similar in both sexes (30.8% in females, 30.3% in males), SEP disparities were stronger in females (32.7% in SEP level 1 vs. 27.7% in SEP level 4; 18.1% relative difference) compared to males (30.6% in SEP level 1 vs. 29.3% in SEP level 4; 4.4% relative difference). The overall prevalence of morbid obesity in this population was 20.8% in 2016. The rate among females was almost double compared to the rate in males (28.1% and 14.6%, respectively). In both sexes, the prevalence of morbid obesity was strongly associated with lower SEP. However, in females, disparities between SEP levels were much stronger (34.3% in SEP level 1 vs. 18.7% in SEP level 4; 83.4% relative difference) compared to SEP-disparities in males (15.7% in SEP level 1 vs. 12.3% in SEP level 4; 27.6% relative difference). Conclusions: The overall prevalence of BMI≥ 30 kg/m2 among adults with diabetes in Israel exceeds 50%; and the prevalence of morbid obesity suggests that 20% meet the BMI-criteria for bariatric surgery. Prevalence rates show major SEP- and sex-disparities; especially strong SEP disparities in morbid obesity among females. These findings highlight the need for greater consideration of different population groups when implementing interventions.

Keywords: diabetes, health disparities, health policy, obesity, socio-economic position

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2593 Dwindling the Stability of DNA Sequence by Base Substitution at Intersection of COMT and MIR4761 Gene

Authors: Srishty Gulati, Anju Singh, Shrikant Kukreti

Abstract:

The manifestation of structural polymorphism in DNA depends on the sequence and surrounding environment. Ample of folded DNA structures have been found in the cellular system out of which DNA hairpins are very common, however, are indispensable due to their role in the replication initiation sites, recombination, transcription regulation, and protein recognition. We enumerate this approach in our study, where the two base substitutions and change in temperature embark destabilization of DNA structure and misbalance the equilibrium between two structures of a sequence present at the overlapping region of the human COMT gene and MIR4761 gene. COMT and MIR4761 gene encodes for catechol-O-methyltransferase (COMT) enzyme and microRNAs (miRNAs), respectively. Environmental changes and errors during cell division lead to genetic abnormalities. The COMT gene entailed in dopamine regulation fosters neurological diseases like Parkinson's disease, schizophrenia, velocardiofacial syndrome, etc. A 19-mer deoxyoligonucleotide sequence 5'-AGGACAAGGTGTGCATGCC-3' (COMT19) is located at exon-4 on chromosome 22 and band q11.2 at the intersection of COMT and MIR4761 gene. Bioinformatics studies suggest that this sequence is conserved in humans and few other organisms and is involved in recognition of transcription factors in the vicinity of 3'-end. Non-denaturating gel electrophoresis and CD spectroscopy of COMT sequences indicate the formation of hairpin type DNA structures. Temperature-dependent CD studies revealed an unusual shift in the slipped DNA-Hairpin DNA equilibrium with the change in temperature. Also, UV-thermal melting techniques suggest that the two base substitutions on the complementary strand of COMT19 did not affect the structure but reduces the stability of duplex. This study gives insight about the possibility of existing structurally polymorphic transient states within DNA segments present at the intersection of COMT and MIR4761 gene.

Keywords: base-substitution, catechol-o-methyltransferase (COMT), hairpin-DNA, structural polymorphism

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2592 Erectile Dysfunction among Bangladeshi Men with Diabetes

Authors: Shahjada Selim

Abstract:

Background: Erectile dysfunction (ED) is an important impediment to quality of life of men. ED is approximate, three times more common in diabetic than non-diabetic men, and diabetic men develop ED earlier than age-matched non-diabetic subjects. Glycemic control and other factors may contribute in developing and or deteriorating ED. Aim: The aim of the study was to determine the prevalence of ED and its risk factors in type 2 diabetic (T2DM) men in Bangladesh. Methods: During 2013-2014, 3980 diabetic men aged 30-69 years were interviewed at the out-patient departments of seven diabetic centers in Dhaka by using the validated Bengali version of the questionnaire of the International index of erectile function (IIEF) for evaluation of baseline erectile function (EF). The indexes indicate a very high correlation between the items and the questionnaire is consistently reliable. Data were analyzed with Chi-squared (χ²) test using SPSS software. P ≤ 0.05 was considered significant. Results: Out of 3790, ED was found in 2046 (53.98%) of T2DM men. The prevalence of ED was increased with age from 10.5% in men aged 30-39 years to 33.6% in those aged over 60 years (P < 0.001). In comparison with patients with reported diabetes lasting ≤ 5 years (26.4%), the prevalence of ED was less than in those with diabetes of 6-11 years (35.3%) and of 12-30 years (42.5%, P <0.001). ED increased significantly in those who had poor glycemic control. The prevalence of ED in patients with good, fair and poor glycemic control was 22.8%, 42.5% and 47.9% respectively (P = 0.004). Treatment modalities (medical nutrition therapy, oral agents, insulin, and insulin plus oral agents) had significant association with ED and its severity (P < 0.001). Conclusion: Prevalence of ED is very high among T2DM men in Bangladesh and can be reduced the burden by improving glycemic status. Glycemic control, duration of diabetes, treatment modalities, increasing age are associated with ED.

Keywords: erectile dysfunction, diabetes, men, Bangladesh

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2591 Recognition of Gene Names from Gene Pathway Figures Using Siamese Network

Authors: Muhammad Azam, Micheal Olaolu Arowolo, Fei He, Mihail Popescu, Dong Xu

Abstract:

The number of biological papers is growing quickly, which means that the number of biological pathway figures in those papers is also increasing quickly. Each pathway figure shows extensive biological information, like the names of genes and how the genes are related. However, manually annotating pathway figures takes a lot of time and work. Even though using advanced image understanding models could speed up the process of curation, these models still need to be made more accurate. To improve gene name recognition from pathway figures, we applied a Siamese network to map image segments to a library of pictures containing known genes in a similar way to person recognition from photos in many photo applications. We used a triple loss function and a triplet spatial pyramid pooling network by combining the triplet convolution neural network and the spatial pyramid pooling (TSPP-Net). We compared VGG19 and VGG16 as the Siamese network model. VGG16 achieved better performance with an accuracy of 93%, which is much higher than OCR results.

Keywords: biological pathway, image understanding, gene name recognition, object detection, Siamese network, VGG

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2590 Interaction Between Gut Microorganisms and Endocrine Disruptors - Effects on Hyperglycaemia

Authors: Karthika Durairaj, Buvaneswari G., Gowdham M., Gilles M., Velmurugan G.

Abstract:

Background: Hyperglycaemia is the primary cause of metabolic illness. Recently, researchers focused on the possibility that chemical exposure could promote metabolic disease. Hyperglycaemia causes a variety of metabolic diseases dependent on its etiologic conditions. According to animal and population-based research, individual chemical exposure causes health problems through alteration of endocrine function with the influence of microbial influence. We were intrigued by the function of gut microbiota variation in high fat and chemically induced hyperglycaemia. Methodology: C57/Bl6 mice were subjected to two different treatments to generate the etiologic-based diabetes model: I – a high-fat diet with a 45 kcal diet, and II - endocrine disrupting chemicals (EDCs) cocktail. The mice were monitored periodically for changes in body weight and fasting glucose. After 120 days of the experiment, blood anthropometry, faecal metagenomics and metabolomics were performed and analyzed through statistical analysis using one-way ANOVA and student’s t-test. Results: After 120 days of exposure, we found hyperglycaemic changes in both experimental models. The treatment groups also differed in terms of plasma lipid levels, creatinine, and hepatic markers. To determine the influence on glucose metabolism, microbial profiling and metabolite levels were significantly different between groups. The gene expression studies associated with glucose metabolism vary between hosts and their treatments. Conclusion: This research will result in the identification of biomarkers and molecular targets for better diabetes control and treatment.

Keywords: hyperglycaemia, endocrine-disrupting chemicals, gut microbiota, host metabolism

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2589 Diabetes and Medical Plant's Treatment: Ethnobotanical Studies Carried out in Morocco

Authors: Jamila Fakchich, Mostafa Jamila Lazaar Elachouri, Lakhder Fakchich, Fatna Ouali, Abd Errazzak Belkacem

Abstract:

Diabetes is a chronic metabolic disease that has a significant impact on the health, quality of life, and life expectancy of patients as well as the health care system. By its nature diabetes, is a multisystem disease with wide-ranging complication that span nearly all region of the body. This epidemic problem, however, is not unique to the industrialized society, but has also hardly struck the developing countries. In Morocco, as developing country, there is an epidemic rise in diabetes, with ensuing concern about the management and control of this disease; it began a chronic burdensome disease of largely middle-aged and elderly people, with a long course and serious complications often resulting in high death-rate, the treatment of diabetes spent vast amount of resources including medicines, diets, physical training. Treatment of this disease is considered problematic due to the lack of effective and safe drugs capable of inducing sustained clinical, biochemical, and histological cure. In Moroccan society, the phytoremedies are some times the only affordable sources of healthcare, particularly for the people in remote areas. In this paper, we present a synthesis work obtained from the ethnobotanical data reported in different specialized journals. A Synthesis of four published ethnobotanical studies that have been carried out in different region of Morocco by different team seekers during the period from 1997 to 2015. Medicinal plants inventoried by different seekers in four Moroccan’s areas have been regrouped and codified, then, Factorial Analysis (FA) and Principal Components Analysis (PCA) are used to analyse the aggregated data from the four studies and plants are classified according to their frequency of use by population. Our work deals with an attempt to gather information on some traditional uses of medicinal plants from different regions of Morocco, also, it was designed to give a set of medicinal plants commonly used by Moroccan people in the treatment of diabetes; In this paper, we intended to provide a basic knowledge about plant species used by Moroccan society for treatment of diabetes. One of the most interesting aspects of this type of works is to assess the relative cultural importance of medicinal plants for specific illnesses and exploring its usefulness in the context of diabetes.

Keywords: Morocco, medicinal plants, ethnobotanical, diabetes, phytoremedies

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2588 Expression Level of Dehydration-Responsive Element Binding/DREB Gene of Some Local Corn Cultivars from Kisar Island-Maluku Indonesia Using Quantitative Real-Time PCR

Authors: Hermalina Sinay, Estri L. Arumingtyas

Abstract:

The research objective was to determine the expression level of dehydration responsive element binding/DREB gene of local corn cultivars from Kisar Island Maluku. The study design was a randomized block design with single factor consist of six local corn cultivars obtained from farmers in Kisar Island and one reference varieties wich has been released by the government as a drought-tolerant varieties and obtained from Cereal Crops Research Institute (ICERI) Maros South Sulawesi. Leaf samples were taken is the second leaf after the flag leaf at the 65 days after planting. Isolation of total RNA from leaf samples was carried out according to the protocols of the R & A-BlueTM Total RNA Extraction Kit and was used as a template for cDNA synthesis. The making of cDNA from total RNA was carried out according to the protocol of One-Step Reverse Transcriptase PCR Premix Kit. Real Time-PCR was performed on cDNA from reverse transcription followed the procedures of Real MODTM Green Real-Time PCR Master Mix Kit. Data obtained from the real time-PCR results were analyzed using relative quantification method based on the critical point / Cycle Threshold (CP / CT). The results of gene expression analysis of DREB gene showed that the expression level of the gene was highest obtained at Deep Yellow local corn cultivar, and the lowest one was obtained at the Rubby Brown Cob cultivar. It can be concluded that the expression level of DREB gene of Deep Yellow local corn cultivar was highest than other local corn cultivars and Srikandi variety as a reference variety.

Keywords: expression, level, DREB gene, local corn cultivars, Kisar Island, Maluku

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2587 Insights into Archaeological Human Sample Microbiome Using 16S rRNA Gene Sequencing

Authors: Alisa Kazarina, Guntis Gerhards, Elina Petersone-Gordina, Ilva Pole, Viktorija Igumnova, Janis Kimsis, Valentina Capligina, Renate Ranka

Abstract:

Human body is inhabited by a vast number of microorganisms, collectively known as the human microbiome, and there is a tremendous interest in evolutionary changes in human microbial ecology, diversity and function. The field of paleomicrobiology, study of ancient human microbiome, is powered by modern techniques of Next Generation Sequencing (NGS), which allows extracting microbial genomic data directly from archaeological sample of interest. One of the major techniques is 16S rRNA gene sequencing, by which certain 16S rRNA gene hypervariable regions are being amplified and sequenced. However, some limitations of this method exist including the taxonomic precision and efficacy of different regions used. The aim of this study was to evaluate the phylogenetic sensitivity of different 16S rRNA gene hypervariable regions for microbiome studies in the archaeological samples. Towards this aim, archaeological bone samples and corresponding soil samples from each burial environment were collected in Medieval cemeteries in Latvia. The Ion 16S™ Metagenomics Kit targeting different 16S rRNA gene hypervariable regions was used for library construction (Ion Torrent technologies). Sequenced data were analysed by using appropriate bioinformatic techniques; alignment and taxonomic representation was done using Mothur program. Sequences of most abundant genus were further aligned to E. coli 16S rRNA gene reference sequence using MEGA7 in order to identify the hypervariable region of the segment of interest. Our results showed that different hypervariable regions had different discriminatory power depending on the groups of microbes, as well as the nature of samples. On the basis of our results, we suggest that wider range of primers used can provide more accurate recapitulation of microbial communities in archaeological samples. Acknowledgements. This work was supported by the ERAF grant Nr. 1.1.1.1/16/A/101.

Keywords: 16S rRNA gene, ancient human microbiome, archaeology, bioinformatics, genomics, microbiome, molecular biology, next-generation sequencing

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2586 Heater and Substrate Profile Optimization for Low Power Portable Breathalyzer to Diagnose Diabetes Mellitus

Authors: Ramji Kalidoss, Snekhalatha Umapathy, V. Dhinakaran, J. M. Mathana

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Chemi-resistive sensors used in breathalyzers have become a hotspot between the international breath research communities. These sensors exhibit a significant change in its resistance depending on the temperature it gets heated thus demanding high power leading to non-portable instrumentation. In this work, numerical simulation to identify the suitable combination of substrate and heater profile using COMSOL multiphysics was studied. Ni-Cr and Pt-100 joule resistive heater with various profiles were studied beneath the square and circular alumina substrates. The temperature distribution was uniform throughout the square substrate with the meander shaped pt100 heater with 48 mW power consumption for 200 oC. Moreover, this heater profile induced minimal stress on the substrate with 0.5 mm thick. A novel Graphene based ternary metal oxide nanocomposite (GO/SnO2/TiO2) was coated on the optimized substrate and heater to elucidate the response of diabetes biomarker (acetone). The sensor exhibited superior gas sensing performance towards acetone in the exhaled breath concentration range for diabetes (0.25 – 3 ppm). These results indicated the importance of substrate and heater properties along with sensing material for low power portable breathalyzers.

Keywords: Breath Analysis, Chemical Sensors, Diabetes Mellitus, Graphene Nanocomposites, Heater, Substrate

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2585 The Contribution of Genetic Polymorphisms of Tumor Necrosis Factor Alpha and Vascular Endothelial Growth Factor into the Unfavorable Clinical Course of Ulcerative Colitis

Authors: Y. I. Tretyakova, S. G. Shulkina, T. Y. Kravtsova, A. A. Antipova, N. Y. Kolomeets

Abstract:

The research aimed to assess the functional significance of tumor necrosis factor-alpha (TNF-α) gene polymorphism at the -308G/A (rs1800629) region and vascular endothelial growth factor A (VEGFA) gene polymorphism at the -634G/C (rs 2010963) region in the development of ulcerative colitis (UC), focusing on patients from the Perm region, Russia. We examined 70 UC patients and 50 healthy donors during the active phase of the disease. Our focus was on TNF-α and VEGF concentration in the blood serum, as well as TNF-α and VEGFA gene polymorphisms at the -308G/А and -634G/C regions, respectively. We found that TNF-α and VEGF levels were significantly higher in patients with severe UC and high endoscopic activity compared to those with milder forms of the disease and low endoscopic activity. These tests could serve as additional non-invasive markers for assessing mucosal damage in the large intestine of UC patients. The frequency of allele variations in the TNF-α gene -308G/A (rs1800629) revealed a significantly higher occurrence of the unfavorable homozygote AA in UC patients compared to donors. Additionally, the major allele G and the allele pair GG were more frequent in patients with mild to moderate disease and 1-2 degree of endoscopic activity than in those with severe UC and 3-4 degree of endoscopic activity (χ2=14.19; p=0.000). We also observed a mutant allele A and the unfavorable homozygote AA associated with severe progressive UC. The occurrence of the mutant allele increased the risk of severe UC by 5 times (OR 5.03; CI 12.07-12.21). We did not find any significant differences in the frequency of the CC homozygote (χ2=1.02; p=0.6; OR=1.32) and the mutant allele C of the VEGFA gene -634G/C (rs 2010963) (χ2=0.01; p=0.913; OR=0.97) between groups of UC patients and healthy individuals. However, we detected that the mutant allele C and the unfavorable homozygote CC of the VEGFA gene were associated with more severe endoscopic changes in the colonic mucosa of UC patients (χ2=25,76; р=0,000; OR=0,15). The presence of the mutant allele increased the risk of severe UC by 6 times (OR 6,78; CI 3,13–14,7). We found a direct correlation between TNF-α and VEGFA gene polymorphisms, increased production of the same factors, disease severity, and endoscopic activity (р=0.000). Therefore, the presence of the mutant allele A and homozygote AA of the TNF-α gene at the -308G/A region and the mutant allele C and homozygote CC of the VEGFA gene at the -634G/C region are associated with risks related to an unfavorable clinical course of UC, frequent recurrences, and rapid progression. These findings should be considered when making prognoses regarding the clinical course of the disease and selecting treatment strategies. The presence of the homozygote AA in the TNF-α gene (rs1800629) is considered a sign of genetic predisposition to UC.

Keywords: gene polymorphism, TNF-α, ulcerative colitis, VEGF

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2584 Prevalence and Predictors of Metabolic Syndrome among Diabetic Clinic Attendees in Sokoto, Nigeria

Authors: Kehinde Joseph Awosan, Balarabe Adami Isah, Edzu Usman Yunusa, Sarafadeen Adeniyi Arisegi, Izuchukwu Obasi, Oluchi Solomon-Anucha

Abstract:

Background: Metabolic syndrome (MetS) is prevalent in patients with diabetes mellitus and a significant risk for major cardiovascular events. Identifying its burden and peculiarities is crucial to preventing complications among those at risk. Aim: This study was conducted to determine the prevalence and predictors of metabolic syndrome among diabetes clinic attendees in Sokoto, Nigeria. Materials and Methods: A cross-sectional study was conducted among 365 patients with type 2 diabetes attending the diabetes clinic of Specialist Hospital, Sokoto, Nigeria. A structured questionnaire was used to obtain data on the respondents’ socio-demographic variables, treatment history, and lifestyle. Blood pressure and anthropometric measurements (including weight, height, and waist circumference) were done for the patients. Likewise, biochemical assessment (including fasting plasma glucose, high-density lipoprotein cholesterol (HDL-c), and triglyceride (TG) was done. Metabolic syndrome was defined according to the National Cholesterol Education Program Adult Treatment Panel III (NCEP ATP III). Data were analyzed using the IBM Statistical Package for Social Sciences (SPSS) version 25. Results: The ages of the patients ranged from 30 to 78 (mean = 50.9 ±11.7) years. The overall prevalence of MetS was 57.3%, with a higher prevalence in females (68.1%) than males (43.0%). The most common components of MetS observed were hypertension (69.2%), and elevated fasting plasma glucose (65.7%); while the predictors of MetS were age > 50 years (OR 6.960, 95% CI: 3.836-12.628, p < 0.001), female sex (OR 2.300, 95% CI: 1.355-3.903, p = 0.002), physical activity (OR 0.214, 95% CI: 0.126-0.363, p < 0.001), and overweight/obesity (OR 3.356, 95% CI: 1.838-6.127, p < 0.001). Conclusion: Metabolic syndrome is prevalent among patients with type 2 diabetes in Sokoto, Nigeria, and the predictors were age > 50 years, female sex, physical activity, and overweight/obesity. Diabetes care providers should screen their patients for MetS to prevent adverse cardiovascular events.

Keywords: prevalence, predictors, metabolic syndrome, diabetes

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2583 Understanding the Influence of Ethnicity on Adherence to Antidiabetic Medication: Meta-Ethnography and Systematic Review

Authors: Rayah Asiri, Anna Robinson-Barella, Adam Todd, Andy Husband

Abstract:

Introduction: A high prevalence of diabetes and diabetes-related complications in ethnic minority communities is of significant concern. Several studies have indicated low adherence rates to antidiabetic medications in ethnic minorities. Poor adherence to antidiabetic medications leads to a higher risk of complications and mortality. This review aims to explore the barriers to and facilitators of adherence to antidiabetic medication among ethnic minority groups in high-income countries. Methods: A comprehensive search of MEDLINE, Embase, CINAHL, and PsycINFO databases for qualitative studies exploring the barriers to or facilitators of adherence to antidiabetic medication in ethnic minority groups were conducted from database inception to March 2022 (PROSPERO CRD42022320681). A quality assessment of the studies was conducted using the Critical Appraisal Skills Programme (CASP) tool. Key concepts and themes from relevant studies were synthesised using a meta-ethnographic approach. Result: A total of 18 studies were included in the review, and three major themes were developed: 1) cultural underpinnings, 2) communication and building relationships, and 3) managing diabetes during holidays. Conclusion: Multiple barriers and facilitators of adherence to antidiabetic medication among ethnic minority people in high-income countries have been identified. A medication adherence intervention focusing on identified barriers to adherence to antidiabetic medication in ethnic minorities may help in improving diabetes outcomes in these groups.

Keywords: medication adherence, diabetes, ethnic minority, barriers, facilitators

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2582 Mathematical Modeling for Diabetes Prediction: A Neuro-Fuzzy Approach

Authors: Vijay Kr. Yadav, Nilam Rathi

Abstract:

Accurate prediction of glucose level for diabetes mellitus is required to avoid affecting the functioning of major organs of human body. This study describes the fundamental assumptions and two different methodologies of the Blood glucose prediction. First is based on the back-propagation algorithm of Artificial Neural Network (ANN), and second is based on the Neuro-Fuzzy technique, called Fuzzy Inference System (FIS). Errors between proposed methods further discussed through various statistical methods such as mean square error (MSE), normalised mean absolute error (NMAE). The main objective of present study is to develop mathematical model for blood glucose prediction before 12 hours advanced using data set of three patients for 60 days. The comparative studies of the accuracy with other existing models are also made with same data set.

Keywords: back-propagation, diabetes mellitus, fuzzy inference system, neuro-fuzzy

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2581 Allele Mining for Rice Sheath Blight Resistance by Whole-Genome Association Mapping in a Tail-End Population

Authors: Naoki Yamamoto, Hidenobu Ozaki, Taiichiro Ookawa, Youming Liu, Kazunori Okada, Aiping Zheng

Abstract:

Rice sheath blight is one of the destructive fungal diseases in rice. We have thought that rice sheath blight resistance is a polygenic trait. Host-pathogen interactions and secondary metabolites such as lignin and phytoalexins are likely to be involved in defense against R. solani. However, to our knowledge, it is still unknown how sheath blight resistance can be enhanced in rice breeding. To seek for an alternative genetic factor that contribute to sheath blight resistance, we mined relevant allelic variations from rice core collections created in Japan. Based on disease lesion length on detached leaf sheath, we selected 30 varieties of the top tail-end and the bottom tail-end, respectively, from the core collections to perform genome-wide association mapping. Re-sequencing reads for these varieties were used for calling single nucleotide polymorphisms among the 60 varieties to create a SNP panel, which contained 1,137,131 homozygous variant sites after filitering. Association mapping highlighted a locus on the long arm of chromosome 11, which is co-localized with three sheath blight QTLs, qShB11-2-TX, qShB11, and qSBR-11-2. Based on the localization of the trait-associated alleles, we identified an ankyryn repeat-containing protein gene (ANK-M) as an uncharacterized candidate factor for rice sheath blight resistance. Allelic distributions for ANK-M in the whole rice population supported the reliability of trait-allele associations. Gene expression characteristics were checked to evaluiate the functionality of ANK-M. Since an ANK-M homolog (OsPIANK1) in rice seems a basal defense regulator against rice blast and bacterial leaf blight, ANK-M may also play a role in the rice immune system.

Keywords: allele mining, GWAS, QTL, rice sheath blight

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2580 Enhanced Kinetic Solubility Profile of Epiisopiloturine Solid Solution in Hipromellose Phthalate

Authors: Amanda C. Q. M. Vieira, Cybelly M. Melo, Camila B. M. Figueirêdo, Giovanna C. R. M. Schver, Salvana P. M. Costa, Magaly A. M. de Lyra, Ping I. Lee, José L. Soares-Sobrinho, Pedro J. Rolim-Neto, Mônica F. R. Soares

Abstract:

Epiisopiloturine (EPI) is a drug candidate that is extracted from Pilocarpus microphyllus and isolated from the waste of Pilocarpine. EPI has demonstrated promising schistosomicidal, leishmanicide, anti-inflammatory and antinociceptive activities, according to in vitro studies that have been carried out since 2009. However, this molecule shows poor aqueous solubility, which represents a problem for the release of the drug candidate and its absorption by the organism. The purpose of the present study is to investigate the extent of enhancement of kinetic solubility of a solid solution (SS) of EPI in hipromellose phthalate HP-55 (HPMCP), an enteric polymer carrier. SS was obtained by the solvent evaporation methodology, using acetone/methanol (60:40) as solvent system. Both EPI and polymer (drug loading 10%) were dissolved in this solvent until a clear solution was obtained, and then dried in oven at 60ºC during 12 hours, followed by drying in a vacuum oven for 4 h. The results show a considerable modification in the crystalline structure of the drug candidate. For instance, X-ray diffraction (XRD) shows a crystalline behavior for the EPI, which becomes amorphous for the SS. Polarized light microscopy, a more sensitive technique than XRD, also shows completely absence of crystals in SS sample. Differential Scanning Calorimetric (DSC) curves show no signal of EPI melting point in SS curve, indicating, once more, no presence of crystal in this system. Interaction between the drug candidate and the polymer were found in Infrared microscopy, which shows a carbonyl 43.3 cm-1 band shift, indicating a moderate-strong interaction between them, probably one of the reasons to the SS formation. Under sink conditions (pH 6.8), EPI SS had its dissolution performance increased in 2.8 times when compared with the isolated drug candidate. EPI SS sample provided a release of more than 95% of the drug candidate in 15 min, whereas only 45% of EPI (alone) could be dissolved in 15 min and 70% in 90 min. Thus, HPMCP demonstrates to have a good potential to enhance the kinetic solubility profile of EPI. Future studies to evaluate the stability of SS are required to conclude the benefits of this system.

Keywords: epiisopiloturine, hipromellose phthalate HP-55, pharmaceuticaltechnology, solubility

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2579 Cadmium Levels in Patients with Type 2 Diabetes Mellitus in Thasala Southern Thailand

Authors: Supabhorn Yimthiang, Wiyada Khanwian

Abstract:

Cadmium is a heavy metal that is important in the environment because it is highly toxic. The incidence and severity of type 2 diabetes mellitus are known to be associated with cadmium. The purpose of this study was to investigate the cadmium levels in patients with type 2 diabetes mellitus at diabetes mellitus clinic, Thasala hospital, Nakhon Si Thummarat, Thailand. The study population was composed of forty five subjects. Among them, twenty two were diabetic patients and twenty three were apparently healthy non-diabetic individual subjects. After an overnight fasting, blood and morning urine samples were collected from each subject to determine fasting blood sugar and cadmium levels in urine, respectively. Systolic and diastolic blood pressure values were measured by aneroid sphygmomanometer. Study approval was taken from the human subject ethics committee of Walailak University. Verbal and written informed consent was taken from all participants. In the study samples, there were 31.8% males and 68.2% females with mean age of 47+10.53 years. The geometric mean of urine cadmium was significantly higher in diabetic patients (1.015 + 0.79 µg/g creatinine) when compared with the healthy subjects (0.395 + 0.53 µg/g creatinine) (P<0.05). This result also showed that urine cadmium excretion in diabetic patients was higher than in healthy subjects by 2.6 times. Moreover, fasting blood sugar (153+47.86 μg/dl) and systolic blood pressure (183.26+17.15 mmHg) of diabetic patients was significantly different when compared with healthy subjects (79+5.38 μg/dl and 112.78+11.32 mmHg, respectively) (P<0.05). Meanwhile, the concentration of cadmium in urine showed positive correlation with fasting plasma glucose (r=0.616) and systolic blood pressure (r=0.487). This preliminary study showed that cadmium might play an important role in the development and pathogenesis of diabetes mellitus in general population. However, these findings require confirmation through additional epidemiological and biological research.

Keywords: blood pressure, cadmium, fasting blood sugar, type 2 diabetes mellitus

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2578 The Generation of Insulin Producing Cells from Human Mesenchymal Stem Cells by miR-375 and Anti-miR-9

Authors: Arefeh Jafarian, Mohammad Taghikani, Saied Abroun, Amir Allahverdi, Masoud Soleimani

Abstract:

Introduction: The miRNAs have key roles in control of pancreatic islet development and insulin secretion. In this regards, current study investigated the pancreatic differentiation of human bone marrow mesenchymal stem cells (hBM-MSCs) by up-regulation of miR-375 and down-regulation of miR-9 by lentiviruses containing miR-375 and anti-miR-9. Findings: After 21 days of induction, islet-like clusters containing insulin producing cells (IPCs) were confirmed by dithizone (DTZ) staining. The IPCs and β cell specific related genes and proteins were detected using qRT-PCR and immunofluorescence on days 7, 14 and 21 of differentiation. Glucose challenge test was performed at different concentrations of glucose as well as extracellular and intracellular insulin and C-peptide were assayed using ELISA kit. In derived IPCs by miR-375 alone are capable to express insulin and other endocrine specific transcription factors, the cells lack the machinery to respond to glucose. The differentiated hMSCs by miR-375 and anti-miR-9 lentiviruses could secrete insulin and c-peptide in a glucose-regulated manner. Conclusion: It was found that over-expression of miR-375 led to a reduction in levels of Mtpn protein in derived IPCs, while treatment with anti-miR-9 following miR-375 over-expression had synergistic effects on MSCs differentiation and insulin secretion in a glucose-regulated manner. The researchers reported that silencing of miR-9 increased OC-2 protein in IPCs that may contribute to the observed glucose-regulated insulin secretion. These findings highlight miRNAs functions in stem cells differentiation and suggest that they could be used as therapeutic tools for gene-based therapy in diabetes mellitus.

Keywords: diabetes, differentiation, MSCs, insulin producing cells, miR-375, miR-9

Procedia PDF Downloads 292
2577 Lucilia Sericata Netrin-A: Secreted by Salivary Gland Larvae as a Potential to Neuroregeneration

Authors: Hamzeh Alipour, Masoumeh Bagheri, Tahereh Karamzadeh, Abbasali Raz, Kourosh Azizi

Abstract:

Netrin-A, a protein identified for conducting commissural axons, has a similar role in angiogenesis. In addition, studies have shown that one of the netrin-A receptors is expressed in the growing cells of small capillaries. It will be interesting to study this new group of molecules because their role in wound healing will become clearer in the future due to angiogenesis. The greenbottle blowfly Luciliasericata (L. sericata) larvae are increasingly used in maggot therapy of chronic wounds. This aim of this was the identification of moleculareatures of Netrin-A in L. sericata larvae. Larvae were reared under standard maggotarium conditions. The nucleic acid sequence of L. sericataNetrin-A (LSN-A) was then identified using Rapid Amplification of cDNA Ends (RACE) and Rapid Amplification of Genomic Ends (RAGE). Parts of the Netrin-A gene, including the middle, 3′-, and 5′-ends were identified, TA cloned in pTG19 plasmid, and transferred into DH5ɑ Escherichia coli. Each part was sequenced and assembled using SeqMan software. This gene structure was further subjected to in silico analysis. The DNA of LSN-A was identified to be 2407 bp, while its mRNA sequence was recognized as 2115 bp by Oligo0.7 software. It translated the Netrin-A protein with 704 amino acid residues. Its molecular weight is estimated to be 78.6 kDa. The 3-D structure ofNetrin-A drawn by SWISS-MODEL revealed its similarity to the Netrin-1 of humans with 66.8% identity. The LSN-A protein conduces to repair the myelin membrane in neuronal cells. Ultimately, it can be an effective candidate in neural regeneration and wound healing. Furthermore, our next attempt is to deplore recombinant proteins for use in medical sciences.

Keywords: maggot therapy, netrin-A, RACE, RAGE, lucilia sericata

Procedia PDF Downloads 79
2576 Mutation Analysis of the ATP7B Gene in 43 Vietnamese Wilson’s Disease Patients

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

Abstract:

Wilson’s disease (WD) is an autosomal recessive disorder of the copper metabolism, which is caused by a mutation in the copper-transporting P-type ATPase (ATP7B). The mechanism of this disease is the failure of hepatic excretion of copper to bile, and leads to copper deposits in the liver and other organs. The ATP7B gene is located on the long arm of chromosome 13 (13q14.3). This study aimed to investigate the gene mutation in the Vietnamese patients with WD, and make a presymptomatic diagnosis for their familial members. Forty-three WD patients and their 65 siblings were identified as having ATP7B gene mutations. Genomic DNA was extracted from peripheral blood samples; 21 exons and exon-intron boundaries of the ATP7B gene were analyzed by direct sequencing. We recognized four mutations ([R723=; H724Tfs*34], V1042Cfs*79, D1027H, and IVS6+3A>G) in the sum of 20 detectable mutations, accounting for 87.2% of the total. Mutation S105* was determined to have a high rate (32.6%) in this study. The hotspot regions of ATP7B were found at exons 2, 16, and 8, and intron 14, in 39.6 %, 11.6 %, 9.3%, and 7 % of patients, respectively. Among nine homozygote/compound heterozygote siblings of the patients with WD, three individuals were determined as asymptomatic by screening mutations of the probands. They would begin treatment after diagnosis. In conclusion, 20 different mutations were detected in 43 WD patients. Of this number, four novel mutations were explored, including [R723=; H724Tfs*34], V1042Cfs*79, D1027H, and IVS6+3A>G. The mutation S105* is the most prevalent and has been considered as a biomarker that can be used in a rapid detection assay for diagnosis of WD patients. Exons 2, 8, and 16, and intron 14 should be screened initially for WD patients in Vietnam. Based on risk profile for WD, genetic testing for presymptomatic patients is also useful in diagnosis and treatment.

Keywords: ATP7B gene, mutation detection, presymptomatic diagnosis, Vietnamese Wilson’s disease

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2575 PMEL Marker Identification of Dark and Light Feather Colours in Local Canary

Authors: Mudawamah Mudawamah, Muhammad Z. Fadli, Gatot Ciptadi, Aulanni’am

Abstract:

Canary breeders have spread throughout Indonesian regions for the low-middle society and become an income source for them. The interesting phenomenon of the canary market is the feather colours become one of determining factor for the price. The advantages of this research were contributed to the molecular database as a base of selection and mating for the Indonesia canary breeder. The research method was experiment with the genome obtained from canary blood isolation. The genome did the PCR amplification with PMEL marker followed by sequencing. Canaries were used 24 heads of light and dark colour feathers. Research data analyses used BioEdit and Network 4.6.0.0 software. The results showed that all samples were amplification with PMEL gene with 500 bp fragment length. In base sequence of 40 was found Cytosine(C) in the light colour canaries, while the dark colour canaries was obtained Thymine (T) in same base sequence. Sequence results had 286-415 bp fragment and 10 haplotypes. The conclusions were the PMEL gene (gene of white pigment) was likely to be used PMEL gene to detect molecular genetic variation of dark and light colour feather.

Keywords: canary, haplotype, PMEL, sequence

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2574 Identification and Characterization of 18S rRNA Gene of Demodex Canis From the Dog Population of Mizoram, India

Authors: Moneesh Thakur, Hridayesh Prasad, Nikitasha Bora, Parimal Roy Choudhary, A. K. Samanta, Sanjeev Kumar

Abstract:

Canine demodicosis is a common parasitic condition which involves dog skin. Demodicosis in dogs is due the prominent growth of Demodex. Out of various canine Demodex spp., Demodex canis is the most often involved species. Canine demodicosis can occur as either a localized or generalized form of demodicosis severely affect the dogs and in non-treated dogs may cause death. This study was planned with the aim to screen and characterize the 18S rRNA gene of isolated Demodex canis. A total of 1200 dogs were screened during this study period. The skin scrapings of all the suspected dogs were examined under a microscope at 100X magnification for the presence of Demodex canis. The skin scrapings positive for Demodex canis were examined using PCR for confirmation. A total of 35 dogs were confirmed a positive result for D. canis based on 18S rRNA gene amplification by PCR. Further, the 18S rRNA gene of isolated Demodex canis was cloned and sequenced for genome analysis. On the sequence analysis, it was found that isolated sequence (GenBank Accession No. MK177513) had close similarity (99.7%) to that of D. canis genotype of China (Accession No. MG372254).

Keywords: PCR, phylogenetic analysis, cloning and sequening, Demodex canis

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2573 Time-Course Lipid Accumulation and Transcript Analyses of Lipid Biosynthesis Gene of Chlorella sp.3 under Nitrogen Limited Condition

Authors: Jyoti Singh, Swati Dubey, Mukta Singh, R. P. Singh

Abstract:

The freshwater microalgae Chlorella sp. is alluring considerable interest as a source for biofuel production due to its fast growth rate and high lipid content. Under nitrogen limited conditions, they can accumulate significant amounts of lipids. Thus, it is important to gain insight into the molecular mechanism of their lipid metabolism. In this study under nitrogen limited conditions, regular pattern of growth characteristics lipid accumulation and gene expression analysis of key regulatory genes of lipid biosynthetic pathway were carried out in microalgae Chlorella sp 3. Our results indicated that under nitrogen limited conditions there is a significant increase in the lipid content and lipid productivity, achieving 44.21±2.64 % and 39.34±0.66 mg/l/d at the end of the cultivation, respectively. Time-course transcript patterns of lipid biosynthesis genes i.e. acetyl coA carboxylase (accD) and diacylglycerol acyltransferase (dgat) showed that during late log phase of microalgae Chlorella sp.3 both the genes were significantly up regulated as compared to early log phase. Moreover, the transcript level of the dgat gene is two-fold higher than the accD gene. The results suggested that both the genes responded sensitively to the nitrogen limited conditions during the late log stage, which proposed their close relevance to lipid biosynthesis. Further, this transcriptome data will be useful for engineering microalgae species by targeting these genes for genetic modification to improve microalgal biofuel quality and production.

Keywords: biofuel, gene, lipid, microalgae

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