Search results for: candidate gene of type-2 diabetes

Authors: Intanri Kurniati, Raja Iqbal Mulya Harahap, Agustyas Tjiptaningrum, Reni Zuraida

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Background: Diabetes mellitus is still increasing and has become a health and social burden in the world. It is known that glycation among various proteins is increased in diabetic patients compared with non-diabetic subjects. Some of these glycated proteins are suggested to be involved in the development and progression of chronic diabetic complications. Among these glycated proteins, glycated hemoglobin (HbA1C) is commonly used as the gold standard index of glycemic control in the clinical setting. HbA1C testing has some methods, and the most commonly used is immunoturbidimetry. This research aimed to compare the HbA1c level between immunoturbidimetry and HbA1C level in T2DM patients. Methods: This research involves 77 patients from Abd Muluk Hospital Bandar Lampung; the patient was asked for consent in this research, then underwent phlebotomy to be tested for HbA1C; the sample was then examined for HbA1C with Turbidimetric Inhibition Immunoassay (TINIA) and High-Performance Liquid Chromatography (HPLC) method. Result: Mean± SD of the samples with the TINIA method was 9.2±1,2; meanwhile, the level HbA1C with the HPLC method is 9.6±1,2. The t-test showed no significant difference between the group subjects. (p<0.05). It was proposed that the two methods have high suitability in testing, and both are eligibly used for the patient. Discussion: There was no significant difference among research subjects, indicating that the high conformity of the two methods is suitable to be used for monitoring patients clinically. Conclusion: There is increasing in HbA1C level in a patient with T2DM measured with HPLC and or Turbidimetric Inhibition Immunoassay (TINIA) method, and there were no significant differences among those methods.

Keywords: diabetes mellitus, glycated albumin, HbA1C, HPLC, immunoturbidimetry

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Authors: Alaa Mostafa, Samuel Dagogo-Jack, Vivian Thieu, Maria Yu, Nan Zhang, Dara Schuster, Luis-Emilio Garcia-Perez

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Dulaglutide (DU), a once-weekly glucagon-like peptide-1 receptor agonist, was studied in the AWARD clinical trial program in adult patients with type 2 diabetes (T2D) and demonstrated significant hemoglobin A1c (A1c) reduction and potential for weight loss. To evaluate the efficacy and safety of DU 1.5 mg and DU 0.75 mg in patients with T2D by baseline A1c <8.5% or ≥8.5%, a post-hoc analysis was conducted on AWARD-1 to -6 and -8 at 6 months. Across 7 studies, 55% to 82% of the DU-treated patients had a baseline A1c <8.5%, and 18% to 45% had a baseline A1c ≥8.5%. The ranges of A1c reductions with baseline A1c <8.5% and ≥8.5%, respectively, were: DU 1.5 mg: -0.67% to -1.25% and -1.22% to -2.37%; DU 0.75 mg: -0.53% to -1.07% and -1.37% to -2.19%. The A1c reduction from the pooled analysis was greater in patients with baseline A1c ≥8.5% than patients with baseline A1c <8.5%, respectively: DU 1.5 mg: -1.86% and -1.02%; DU 0.75 mg: -1.75% and -0.83%. DU treatments were well tolerated among baseline A1c subgroups. Across the AWARD program, DU 1.5 mg and DU 0.75 mg demonstrated significant A1c reduction in both subgroups with an acceptable safety profile. Compared to patients with baseline A1c <8.5%, patients with baseline A1c ≥8.5% had greater A1c reduction. Disclosures: This study was supported and conducted by Eli Lilly and Company, Indianapolis, IN, USA.

Keywords: A1c reduction, dulaglutide, type 2 diabetes, weight loss

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Authors: A. Porshnev, A. Zaporozhtchuk

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Motivation regarded as a key factor of labor turnover, is especially important for volunteers working on an altruistic basis in NGO. Despite the motivational letter, candidate selection depends on the impression of the selection committee, which can be subject to human bias. We expect that structured and unstructured information provided in motivation letters could be used to improve candidate selection procedures. In our paper, we perform qualitative and quantitative analysis of 2280 motivation letters, create logistic regression, and build a decision tree to improve selection procedures. Our analysis showed that motivation factors are significant and enable human resources department to forecast labor turnover and provide extra information to demographic, professional and timing questions. In spite of the average level of accuracy the model demonstrates the selection procedures of company of under consideration can be improved. We also discuss interrelation between answers to open and closed motivation questions, recommend changes in motivational letter templates to ensure more relevant information about applicants and further steps to create more accurate model.

Keywords: decision trees, logistic regression, model, motivational letter, non-governmental organization, retention, turnover

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Authors: Abdullah S. Alghamdi, Khaled Alghamdi, Richard O. Jenkins, Parvez I. Haris

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Background: Quantity and quality of sleep have been reported to be significant risk factors for obesity and development of metabolic disorders such as type 2 diabetes mellitus (T2DM). The relationship between diabetes and sleep quantity was reported to be U-shaped, which means increased or decreased sleeping hours can increase the risk of diabetes. The plasma glucagon levels were found to continuously decrease during night-time sleep in healthy individuals, independently of blood glucose and insulin levels. The disturbance of the circadian rhythm is also important and has been linked with an increased the chance of diabetes incidence. There is a lack of research on sleep patterns on Saudis with T2DM and how this is affected by Ramadan fasting. Aim: To assess the sleeping patterns of Saudis with T2DM (before, during, and after Ramadan), using two different techniques and relate this to their HbA1c levels. Method: This study recruited 82 Saudi with T2DM, who chose to fast during Ramadan, from the Endocrine and Diabetic Centre of Al Iman General Hospital, Riyadh, Saudi Arabia. Ethical approvals for the study were obtained from De Montfort University and Saudi Ministry of Health. Their sleeping patterns were assessed by a self-administered questionnaire (before, during, and after Ramadan). The assessment included the daily total sleeping hours (DTSH), and total night-time sleeping hours (TNTSH) of the participants. In addition, sleeping patterns of 36 patients, randomly selected from the 82 participants, were further tracked during and after Ramadan by using Fitbit Flex 2™ accelerometer. Blood samples were collected in each period for measuring HbA1c. Results: Questionnaire analysis revealed that the sleeping patterns significantly changed between the periods, with shorter hours during Ramadan (P < 0.001 for DTSH, and P < 0.001 for TNTSH). These findings were confirmed by the Fitbit data, which also indicated significant shorter sleeping hours for the DTSH, and the TNTSH during Ramadan (P < 0.001 and P < 0.001, respectively). Although there were no significant correlations between the questionnaire and Fitbit data, the TNTSH were shorter among the participants in all periods by both techniques. The mean HbA1c significantly varied between periods, with lowest level during Ramadan. Although the statistical tests did not show significant variances in the mean HbA1c between the groups of participants regarding their hours of sleeping, the lowest mean HbA1c was observed in the group of participants who slept for 6-8 hours and had longer night-time sleeping hours. Conclusion: A short sleep duration, and absence of night-time sleep were significantly observed among the majority of the study population during Ramadan, which could suppress the full benefits of Ramadan fasting for diabetic patients. This study showed that there is a good agreement between the findings of the questionnaire and the Fitbit device for evaluating sleeping patterns in a Saudi population. A larger study is needed in the future to investigate the impact of Ramadan fasting on sleep quality and quantity and its relationship with health and disease.

Keywords: Diabetes, Fasting, Fitbit, HbA1c, IPAQ, Ramadan, Sleep

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Authors: Fitri Rahmi Fadhilah, Edhyana Sahiratmadja, Ani Melani Maskoen, Ratu Safitri, Supartini Syarif, Herman Susanto

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Cervical cancer is the second most common cancer in women after breast cancer. In Indonesia, the incidence of cervical cancer cases is estimated at 25-40 per 100,000 women per year. Human papillomavirus (HPV) infection is a major cause of cervical cancer, and HPV-16 is the most common genotype that infects the cervical tissue. The major late protein L1 may be associated with infectivity and pathogenicity and its variation can be used to classify HPV isolates. The aim of this study was to determine the phylogenetic tree of HPV 16 L1 gene from cervical cancer patient isolates in Bandung. After confirming HPV-16 by Linear Array Genotyping Test, L1 gene was amplified using specific primers and subject for sequencing. Phylogenetic analysis revealed that HPV 16 from Bandung was in the subgroup of Asia and East Asia, showing the close host-agent relationship among the Asian type.

Keywords: L1 HPV 16, cervical cancer, bandung, phylogenetic

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Authors: Rayhana Begum, HongBin Wang, Nur Alam Siddiquee, Md.Yasin Ahmed

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Biotin treatment has significant effects on blood glucose, and pharmacological doses of biotin improve hyperglycemia. The present study was aimed to investigate the efficacy and safety of biotin in combination with canagliflozin in improving glycemic control on High Fat Diet-induced diabetes in Rats. Thirty male rats were divided into five groups (six rats /group): control, high fat diet (HFD), canagliflozin (CAG), biotin (BIO), and CAG + BIO. The treatments with CAG and /or BIO significantly reduced the body weight gain, blood glucose and HbA1c levels, whereas CAG in combination with BIO revealed greater glycemic improvement than CAG monotherapy. The treatment with CAG and /or BIO causes significant change in lipid profile and CK level while the treatment with CAG in combination with BIO showed better results as compared with CAG monotherapy. Furthermore, combination of biotin with CAG improved the pancreatic and cardiac damage when compared with other treated groups.

Keywords: canagliflozin, biotin, HbA1c, lipid profile

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Authors: J. K Adedeji, O. H Olowomofe, C. O Alo, S.T Ijatuyi

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The issue of high blood sugar level, the effects of which might end up as diabetes mellitus, is now becoming a rampant cardiovascular disorder in our community. In recent times, a lack of awareness among most people makes this disease a silent killer. The situation calls for urgency, hence the need to design a device that serves as a monitoring tool such as a wrist watch to give an alert of the danger a head of time to those living with high blood glucose, as well as to introduce a mechanism for checks and balances. The neural network architecture assumed 8-15-10 configuration with eight neurons at the input stage including a bias, 15 neurons at the hidden layer at the processing stage, and 10 neurons at the output stage indicating likely symptoms cases. The inputs are formed using the exclusive OR (XOR), with the expectation of getting an XOR output as the threshold value for diabetic symptom cases. The neural algorithm is coded in Java language with 1000 epoch runs to bring the errors into the barest minimum. The internal circuitry of the device comprises the compatible hardware requirement that matches the nature of each of the input neurons. The light emitting diodes (LED) of red, green, and yellow colors are used as the output for the neural network to show pattern recognition for severe cases, pre-hypertensive cases and normal without the traces of diabetes mellitus. The research concluded that neural network is an efficient Accu-Chek design tool for the proper monitoring of high glucose levels than the conventional methods of carrying out blood test.

Keywords: Accu-Check, diabetes, neural network, pattern recognition

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Authors: Choumous Mannoubi, Dahlia Kairy, Brigitte Vachon

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The use of technology allows clinicians to provide an individualized approach in a cost-effective manner and to reach a broader client base more easily. Such interventions can be effective in ensuring self-management and follow-up of people with diabetes, reducing the risk of complications by improving accessibility to care services, and better adherence to health recommendations. Consideration of users' opinions and fears to inform the design and implementation stages of these telehealth services seems to be essential to improve their acceptance and usability. The objective of this study is to describe the telepractice of nutritionists supporting the therapeutic management of diabetic patients and document the functional requirements of nutritionists for the design of a tele-nutrition platform. To best identify the requirements and constraints of nutritionists, we conducted individual semi-structured interviews with 10 nutritionists who offered tele-nutrition services. Using a qualitative design with a descriptive approach based on the Nutrition Care Process Model (mNCP) framework, we explored in depth the state of nutritionists' telepractice in public and private health care settings, as well as their requirements for teleconsultation. Qualitative analyses revealed that nutritionists primarily used telephone calls during the COVID 19 pandemic to provide teleconsultations. Nutritionists identified the following important features for the design of a tele-nutrition platform: it should support interprofessional collaboration, allow for the development and monitoring of a care plan, integrate with the existing IT environment, be easy to use, accommodate different levels of patient literacy, and allow for easy sharing of educational materials to support nutrition education.

Keywords: telehealth, nutrition, diabetes, telenutrition, teleconsultation, telemonitoring

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Authors: Anes Enakoa, Yawei Liang

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Question Answering (QA) system is one of the most important and demanding tasks in the field of Natural Language Processing (NLP). In QA systems, the answer generation task generates a list of candidate answers to the user's question, in which only one answer is correct. Answer selection is one of the main components of the QA, which is concerned with selecting the best answer choice from the candidate answers suggested by the system. However, the selection process can be very challenging especially in Arabic due to its particularities. To address this challenge, an approach is proposed to answer questions with multiple answer choices for Arabic QA systems based on Textual Entailment (TE) recognition. The developed approach employs a Support Vector Machine that considers lexical, semantic and syntactic features in order to recognize the entailment between the generated hypotheses (H) and the text (T). A set of experiments has been conducted for performance evaluation and the overall performance of the proposed method reached an accuracy of 67.5% with C@1 score of 80.46%. The obtained results are promising and demonstrate that the proposed method is effective for TE recognition task.

Keywords: information retrieval, machine learning, natural language processing, question answering, textual entailment

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Authors: Hannah Rachel Vasanthi, Paomipem Phazang, Veereshkumar, Sali, Ramesh Parjapath, Sangeetha Marimuthu Kannan

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Garcinia indica (G. indica) fruit rind extract commonly used in South Indian culinary and Indian System of medicines is reported to exhibit various biological activities. The present study envisages the influence of the phytoconstituents in G. indica extract (Vrikshamla capsules- a herbal supplement) on diabetic condition. The condition of type 2 diabetes was triggered in experimental animals by feeding high fat diet for 8 weeks followed by a sub-diabetogenic dose of 35mg/kg bw of streptozotocin intraperitoneally. Oral supplementation of the extract at two doses (100 and 200 mg/kg body weight) for 14 days reduced hyperglycemia, hypercholesterolemia and dyslipidemia (p< 0.001). Pathophysiological changes of obesity and diabetes associated complications majorly mediated by oxidative stress were analyzed by measuring the markers of oxidative stress such as lipid peroxidation, enzymatic (SOD, Catalase, GPx) and non-enzymatic markers (GSH). Conspicuous changes markers were noticed in diabetic condition which was reverted by the G. indica extract. Screening the extract by AccuTOF-DART (MS) revealed the presence of hydroxycitric acid and garcinol in abundant quantity which probably has influenced the biological activity. This was also corroborated through docking studies of hydroxycitric acid and garcinol both individually and synergistically with the antioxidant proteins. Altogether, hydroxycitric acid and garcinol present in G. indica fruit extract alleviates the pathophysiological conditions such as hyperglycemia, dyslipidemia, insulin resistance and oxidative stress mediated by diabesity.

Keywords: antioxidants , diabesity, hydroxycitric acid, garcinol, Garcinia indica, sreptozotocin

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Authors: D. J. Kalita

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Cancer is one of the prime causes of early death worldwide. Mutation of the gene involve in DNA repair and damage, like BRCA2 (Breast cancer gene two) genes, can be detected efficiently by PCR-RFLP to early breast cancer diagnosis and adopt the suitable method of treatment. Host Defense Peptide can be used as blueprint for the design and synthesis of novel anticancer drugs to avoid the side effect of conventional chemotherapy and chemo resistance. The change at nucleotide position 392 of a -› c in the cancer sample of dog mammary tumour at BRCA2 (exon 7) gene lead the creation of a new restriction site for SsiI restriction enzyme. This SNP may be a marker for detection of canine mammary tumour. Support vector machine (SVM) algorithm was used to design and predict the anticancer peptide from the mature functional peptide. MTT assay of MCF-7 cell line after 48 hours of post treatment showed an increase in the number of rounded cells when compared with untreated control cells. The ability of the synthesized peptide to induce apoptosis in MCF-7 cells was further investigated by staining the cells with the fluorescent dye Hoechst stain solution, which allows the evaluation of the nuclear morphology. Numerous cells with dense, pyknotic nuclei (the brighter fluorescence) were observed in treated but not in control MCF-7 cells when viewed using an inverted phase-contrast microscope. Thus, PCR-RFLP is one of the attractive approach for early diagnosis, and synthetic cationic peptide can be used for the treatment of canine mammary tumour.

Keywords: cancer, cationic peptide, host defense peptides, Breast cancer genes

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Authors: Leona Ondrejikova, Martin Hill, Antonin Parizek

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The incidence of gestational diabetes mellitus (GDM) is higher in women predisposed to developing intrahepatic cholestasis of pregnancy (ICP). Both diseases are associated with altered steroidogenesis when compared with none-ICP controls. However, the effect of GDM on circulating steroids in ICP patients remains unclear. The question remains, whether the levels of circulating steroids differ between ICP patients with and without GDM. In total 10 ICP patients without GDM (ICP+GDM-), 7 ICP patients with GDM (ICP+GDM+), and 15 controls (ICP-GDM-) were monitored during late gestation, at labor, and during three periods postpartum (day 5, week 3, and week 6 postpartum) (Šimják et al., 2018). The relationships between steroid profiles and patients’ status were evaluated using the ANOVA model consisting of subject factor, between-subject factors Group (ICP+GDM+, ICP+GDM-, ICP-GDM-), gestational age at the diagnosis of ICP and gestational age at labor, and within-subject factor Stage and ICP × Stage interaction. The levels of the C21 and C19 Δ5 steroids and 5α/β-reduced C19 steroids were highest in ICP+GDM+, while those for the ICP-GDM-, and ICP+GDM- groups were lower. In the C21 Δ4 steroids and their 5α/β-reduced metabolites, the steroid levels were highest in the ICP+GDM-, intermediate in the ICP-GDM- and lowest in the ICP+GDM+ group. This higher concentration in ICP+GDM- group may be of importance as the 5α-pregnane-3α,20α-diol disulfate, is considered as the substance inducing ICP. In general, these data show that the comorbidity with GDM substantially changes the steroidome in ICP patients towards the higher activity of steroid CYP17A1 lyase step in adrenal zona reticularis reduced CYP17A1 hydroxylase step in zona fasciculata. This is consistent with our previously published hypothesis about the critical role of maternal zona reticularis in the pathophysiology of ICP. Our present data also indicate that the comorbidity with GDM might moderate the gravity of the ICP in this way.

Keywords: CYP17A1, GC-MS, gestational diabetes mellitus, intrahepatic cholestasis of pregnancy

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Authors: Natesan Balasubramanian, Palpandi Pounpandi, Venkatraman Thamil Priya, Vellasamy Shanmugaiah, Karubbiah Balakrishnan, Mandayam Anandam Thirunarayan

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Streptococcus agalactiae is commonly known as Group B Streptococcus (GBS) and it is the most common cause of life-threatening bacterial infection. GBS first considered as a veterinary pathogen causing mastitis in cattle later becomes a human pathogen for severe neonatal infections. In this present study, a total of 20 new clinical isolates of S. agalactiae were collected from male (6) and female patient (14) with different age group. The isolates were from Urinary tract infection (UTI), blood, pus and eye ulcer. All the 20 S. agalactiae isolates has clear hemolysis properties on blood agar medium and were identified by serogrouping and MALTI-TOF-MS analysis. Antibiotic susceptibility/resistance test was performed for 20 S. agalactiae isolates, further phenotypic resistance pattern was observed for tetracycline, vancomycin, ampicillin and penicillin. Genotypically we found two antibiotic resistance genes such as Betalactem antibiotic resistance gene (Tem) (70%) and tetracycline resistance gene Tet(O) 15% in our isolates. Six virulence factors encoding genes were performed by PCR in twenty GBS isolates, cfb gene (100%), followed by, cylE(90.47%), lmp(85.7%), bca(71.42%), rib (38%) and low frequency in bac gene (4.76%) were determined. Most of the S. agalactiae isolates produced strong biofilm in the polystyrene surface (hydrophobic), and low-level biofilm formation was found in glass tube (hydrophilic) surface. lytR is secreted protein and localized in bacterial cell wall, extra cellular membrane, and cytoplasm. In silico docking studies were performed for lytR protein with four antibiofilm compounds, including a peptide (PR39) with the docking study showed peptide has strong interaction followed by ellagic acid and interaction length is 2.95, 2.97 and 2.95 A°. In ligand EGCGO10 and O11 two atoms intract with lytR (Leu271), with binding bond affinity length is 3.24 and 3.14. The aminoacid Leu 271 is act as an impartant aminoacid, since ellagic acid and EGCG interact with same aminoacid.

Keywords: antibiotics, biofilms, clinical isolates, S. agalactiae, virulence

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Authors: Paula I. Buonfiglio, Carlos David Bruque, Lucia Salatino, Vanesa Lotersztein, Sebastián Menazzi, Paola Plazas, Ana Belén Elgoyhen, Viviana Dalamón

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Hearing loss (HL) is the most prevalent sensorineural disorder affecting about 10% of the global population, with more than half due to genetic causes. About 1 in 500-1000 newborns present congenital HL. Most of the patients are non-syndromic with an autosomal recessive mode of inheritance. To date, more than 100 genes are related to HL. Therefore, the Whole-exome sequencing (WES) technique has become a cost-effective alternative approach for molecular diagnosis. Nevertheless, new challenges arise from the detection of novel variants, in particular missense changes, which can lead to a spectrum of genotype-to-phenotype correlations, which is not always straightforward. In this work, we aimed to identify the genetic causes of HL in isolated and familial cases by designing a multistep approach to analyze target genes related to hearing impairment. Moreover, we performed in silico and in vivo analyses in order to further study the effect of some of the novel variants identified in the hair cell function using the zebrafish model. A total of 650 patients were studied by Sanger Sequencing and Gap-PCR in GJB2 and GJB6 genes, respectively, diagnosing 15.5% of sporadic cases and 36% of familial ones. Overall, 50 different sequence variants were detected. Fifty of the undiagnosed patients with moderate HL were tested for deletions in STRC gene by Multiplex ligation-dependent probe amplification technique (MLPA), leading to 6% of diagnosis. After this initial screening, 50 families were selected to be analyzed by WES, achieving diagnosis in 44% of them. Half of the identified variants were novel. A missense variant in MYO6 gene detected in a family with postlingual HL was selected to be further analyzed. A protein modeling with AlphaFold2 software was performed, proving its pathogenic effect. In order to functionally validate this novel variant, a knockdown phenotype rescue assay in zebrafish was carried out. Injection of wild-type MYO6 mRNA in embryos rescued the phenotype, whereas using the mutant MYO6 mRNA (carrying c.2782C>A variant) had no effect. These results strongly suggest the deleterious effect of this variant on the mobility of stereocilia in zebrafish neuromasts, and hence on the auditory system. In the present work, we demonstrated that our algorithm is suitable for the sequential multigenic approach to HL in our cohort. These results highlight the importance of a combined strategy in order to identify candidate variants as well as the in silico and in vivo studies to analyze and prove their pathogenicity and accomplish a better understanding of the mechanisms underlying the physiopathology of the hearing impairment.

Keywords: diagnosis, genetics, hearing loss, in silico analysis, in vivo analysis, WES, zebrafish

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

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

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

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Authors: Savjot Kaur, Mridula Mahajan, AJS Bhanwer, Santokh Singh, Kawaljit Matharoo

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Background: Disorders of lipid metabolism and genetic predisposition are CAD risk factors. ApoA1 is the apolipoprotein component of anti-atherogenic high density lipoprotein (HDL) particles. The protective action of HDL and ApoA1 is attributed to their central role in reverse cholesterol transport (RCT). Aim: This study was aimed at identifying sequence variations in ApoA1 (-75G>A) and its association with serum ApoA1 levels. Methods: A total of 300 CAD patients and 300 Normal individuals (controls) were analyzed. PCR-RFLP method was used to determine the DNA polymorphism in the ApoA1 gene, PCR products digested with restriction enzyme MspI, followed by Agarose Gel Electrophoresis. Serum apolipoprotein A1 concentration was estimated with immunoturbidimetric method. Results: Deviation from Hardy- Weinberg Equilibrium (HWE) was observed for this gene variant. The A- allele frequency was higher among Coronary Artery disease patients (53.8) compared to controls (45.5), p= 0.004, O.R= 1.38(1.11-1.75). Under recessive model analysis (AA vs. GG+GA) AA genotype of ApoA1 G>A substitution conferred ~1 fold increased risk towards CAD susceptibility (p= 0.002, OR= 1.72(1.2-2.43). With serum ApoA1 levels < 107 A allele frequency was higher among CAD cases (50) as compared to controls (43.4) [p=0.23, OR= 1.2(0.84-2)] and there was zero % occurrence of A allele frequency in individuals with ApoA1 levels > 177. Conclusion: Serum ApoA1 levels were associated with ApoA1 promoter region variation and influence CAD risk. The individuals with the APOA1 -75 A allele confer excess hazard of developing CAD as a result of its effect on low serum concentrations of ApoA1.

Keywords: apolipoprotein A1 (G>A) gene polymorphism, coronary artery disease (CAD), reverse cholesterol transport (RCT)

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Authors: Mehrnoosh Azimi Sanavi, Hamed Ghazvini, Mehryar Zargari, Hossein Ghalehnoei, Zahra Hosseini-khah

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Objectives: Calcium Voltage-Gated Channel Subunit Alpha1 C (CACNA1C) is one of the most important genes associated with schizophrenia. Methods: 45 male Wistar rats were divided into 5 groups: saline, control, ketamine, clozapine, and risperidone. Animals in ketamine, risperidone, and clozapine groups received ketamine (30 mg/ kg-i.p.) for 10 days. After the last injection of ketamine, we started injecting clozapine (7.5 mg/kg-i.p.) risperidone (1 mg/kg-i.p.) for up to 28 days. Twenty-four hours after the last injection, open field, social interaction, and elevated plus-maze tests, and gene expression in the hippocampus were performed. Results: The results of the social interaction test revealed a significant decrease in cumulative time with ketamine compared with the saline group and an increase with clozapine and risperidone compared with the ketamine group. Moreover, results from the elevated plus-maze test demonstrated a critical decrease in open-arm time and an increase in close-arm time with ketamine compared with saline, as well as an increase in open-arm time with risperidone compared with ketamine. Further results revealed a significant increase in rearing and grooming with ketamine compared to saline, as well as a decrease with risperidone and clozapine compared to ketamine. There were no significant differences in CACNA1C gene expression between groups in the rat hippocampus. In brief, the results of this study indicated that clozapine and risperidone could partially improve cognitive impairments in the rat. However, our findings demonstrated that this treatment is not related to CACNA1C gene expression.

Keywords: schizophrenia, ketamine, clozapine, risperidone

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Authors: Sarwan Dhir, Suma Basak, Dipika Parajulee

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Alfalfa is renowned for its nutritional and biopharmaceutical value as a perennial forage legume. However, establishing a rapid plant regeneration protocol using somatic embryogenesis and efficient transformation frequency are the crucial prerequisites for gene editing in alfalfa. This study was undertaken to establish and improve the protocol for somatic embryogenesis and subsequent plant regeneration. The experiments were conducted in response to natural sensitivity using various antibiotics such as cefotaxime, carbenicillin, gentamycin, hygromycin, and kanamycin. Using 3-week-old leaf tissue, somatic embryogenesis was initiated on Gamborg’s B5 basal (B5H) medium supplemented with 3% maltose, 0.9µM Kinetin, and 4.5µM 2,4-D. Embryogenic callus (EC) obtained from the B5H medium exhibited a high rate of somatic embryo formation (97.9%) after 3 weeks when the cultures were placed in the dark. Different developmental stages of somatic embryos and cotyledonary stages were then transferred to Murashige and Skoog’s (MS) basal medium under light, resulting in a 94% regeneration rate of plantlets. Our results indicate that leaf segments can grow (tolerate) up to 450 mg/L of cefotaxime and 400 mg/L of carbenicillin in the culture medium. However, the survival threshold for hygromycin at 12.5 mg/L, kanamycin at 250 mg/L, gentamycin at 50 mg/L, and timentin (300 mg/L). The experiment to improve the protocol for achieving efficient transient gene expression in alfalfa through genetic transformation with the Agrobacterium tumefaciens pCAMBIA1304 vector was also conducted. The vector contains two reporter genes such as β-glucuronidase (GUS) and green fluorescent protein (GFP), along with a selectable hygromycin B phosphotransferase gene (HPT), all driven under the CaMV 35s promoter. Various transformation parameters were optimized using 3-week-old in vitro-grown plantlets. The different parameters such as types of explant, leaf ages, preculture days, segment sizes, wounding types, bacterial concentrations, infection periods, co-cultivation periods, different concentrations of acetosyringone, silver nitrate, and calcium chloride were optimized for transient gene expression. The transient gene expression was confirmed via histochemical GUS and GFP visualization under fluorescent microscopy. The data were analyzed based on the semi-quantitative observation of the percentage and number of blue GUS spots on different days of agro-infection. The highest percentage of GUS positivity (76.2%) was observed in 3-week-old leaf segments wounded using a scalpel blade of 11 size- after 3 days of post-incubation at a bacterial concentration of 0.6, with 2 days of preculture, 30 min of bacterial-leaf segment co-cultivation, with the addition of 150 µM acetosyringone, 4 mM calcium chloride, and 75 µM silver nitrate. Our results suggest that various factors influence T-DNA delivery in the Agrobacterium-mediated transformation of alfalfa. The stable gene expression in the putative transgenic tissue was confirmed using PCR amplification of both marker genes, indicating that gene expression in explants was not solely due to Agrobacterium, but also from transformed cells. The improved protocol could be used for generating transgenic alfalfa plants using genome editing techniques such as CRISPR/Cas9.

Keywords: Medicago sativa l. (Alfalfa), agrobacterium tumefaciens, β-glucuronidase, green fluorescent protein, transient gene

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Authors: Mafalda Moreira, Diana Alba, Inês Paiva Ferreira, Rita Calejo, Ana Rita Soares, Leonilde Machado

Abstract:

Intellectual disability (ID) is characterized by deficits in intellectualfunctioningassociatedwithalterations in the adaptive behaviour, whose onset is inthedevelopmentalperiod. Itaffects 3% of the population, ofwhich 10% have a geneticaetiology. One of those causes isAlwadeiSyndrome, with 3 cases describedworldwide. It results from a homozygous nonsense mutation in theRUSC2 gene andisassociatedwithintellectualdisabilityanddysmorphic facialfeatures. Theauthorsreportthe case of a 5-year-old-boy, born to a healthymotherafter a full-termuneventfulpregnancy, thatwasreferred to Neurodevelopmentalconsultationdue toglobal developmentaldelay. Familyhistoryrevealedlearningdifficulties in the paternal brotherhood. Milddismorphicfeatureswereevidentsuch as darkinfraorbitalregion, low-set ears, beakednose, retrognathism, high-archedpalateandjointhyperlaxity. WechslerIntelligenceScale for Children III fullscaleIQ quoted 61. Karyotypeandchromosomalmicroarrayanalysiswerenormal, as well as the fragile X molecular study. DNA sequencingwasthenperformedandallowedtheidentificationof amutation in the RUSC2 gene. Theetiologicaldiagnosisof ID remains unknown in up to 80% of cases, creatinguncertainty in children’sfamilies. Theadvances in DNA sequencingtechnologieshaveincreasedourknowledgeofthegeneticdiseasesinvolved, as theAlwadeisyndromewasonlydescribedsince 2016. Thegeneticdiagnosisof ID allowsfamilygeneticcounselingandenablesthedevelopmentof target therapeutic approaches.

Keywords: intellectual disability, genetic aetiology, alwadei syndrome, RUSC2

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Authors: Swati Srivastava, Mattia Lauriola, Yuval Gilad, Adi Kimchi, Yosef Yarden

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The glucocorticoid receptor (GR) has been proposed to play important, but incompletely understood roles in cancer. Glucocorticoids (GCs) are widely used as co-medication of various carcinomas, due to their ability to reduce the toxicity of chemotherapy. Furthermore, GR antagonism has proven to be a strategy to treat triple negative breast cancer and castration-resistant prostate cancer. These observations suggest differential GR involvement in cancer subtypes. The goal of our study has been to elaborate the current understanding of GR signaling in tumor progression and metastasis. Our study involves two cellular models, non-tumorigenic breast epithelial cells (MCF10A) and Ewing sarcoma cells (CHLA9). In our breast cell model, the results indicated that the GR agonist dexamethasone inhibits EGF-induced mammary cell migration, and this effect was blocked when cells were stimulated with a GR antagonist, namely RU486. Microarray analysis for gene expression revealed that the mechanism underlying inhibition involves dexamenthasone-mediated repression of well-known activators of EGFR signaling, alongside with enhancement of several EGFR’s negative feedback loops. Because GR mainly acts primarily through composite response elements (GREs), or via a tethering mechanism, our next aim has been to find the transcription factors (TFs) which can interact with GR in MCF10A cells.The TF-binding motif overrepresented at the promoter of dexamethasone-regulated genes was predicted by using bioinformatics. To validate the prediction, we performed high-throughput Protein Complementation Assays (PCA). For this, we utilized the Gaussia Luciferase PCA strategy, which enabled analysis of protein-protein interactions between GR and predicted TFs of mammary cells. A library comprising both nuclear receptors (estrogen receptor, mineralocorticoid receptor, GR) and TFs was fused to fragments of GLuc, namely GLuc(1)-X, X-GLuc(1), and X-GLuc(2), where GLuc(1) and GLuc(2) correspond to the N-terminal and C-terminal fragments of the luciferase gene.The resulting library was screened, in human embryonic kidney 293T (HEK293T) cells, for all possible interactions between nuclear receptors and TFs. By screening all of the combinations between TFs and nuclear receptors, we identified several positive interactions, which were strengthened in response to dexamethasone and abolished in response to RU486. Furthermore, the interactions between GR and the candidate TFs were validated by co-immunoprecipitation in MCF10A and in CHLA9 cells. Currently, the roles played by the uncovered interactions are being evaluated in various cellular processes, such as cellular proliferation, migration, and invasion. In conclusion, our assay provides an unbiased network analysis between nuclear receptors and other TFs, which can lead to important insights into transcriptional regulation by nuclear receptors in various diseases, in this case of cancer.

Keywords: epidermal growth factor, glucocorticoid receptor, protein complementation assay, transcription factor

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Authors: Anju Joshi, C. N. Tharamani

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Diabetes is one of the leading cause of death at present and remains an important concern as the prevalence of the disease is increasing at an alarming rate. Therefore, it is crucial to diagnose the accurate levels of glucose for developing an efficient therapeutic for diabetes. Due to the availability of convenient and compact self-testing, continuous monitoring of glucose is feasible nowadays. Enzyme based electrochemical sensing of glucose is quite popular because of its high selectivity but suffers from drawbacks like complicated purification and immobilization procedures, denaturation, high cost, and low sensitivity due to indirect electron transfer. Hence, designing a robust enzyme free platform using transition metal oxides remains crucial for the efficient and sensitive determination of glucose. In the present work, manganese doped nickel oxide nanoparticles (Mn-NiO) has been synthesized onto the surface of multiwalled carbon nanotubes using a simple microwave assisted approach for non-enzymatic electrochemical sensing of glucose. The morphology and structure of the synthesized nanostructures were characterized using scanning electron microscopy (SEM) and X-Ray diffraction (XRD). We demonstrate that the synthesized nanostructures show enormous potential for electrocatalytic oxidation of glucose with high sensitivity and selectivity. Cyclic voltammetry and square wave voltammetry studies suggest superior sensitivity and selectivity of Mn-NiO decorated carbon nanotubes towards the non-enzymatic determination of glucose. A linear response between the peak current and the concentration of glucose has been found to be in the concentration range of 0.01 μM- 10000 μM which suggests the potential efficacy of Mn-NiO decorated carbon nanotubes for sensitive determination of glucose.

Keywords: diabetes, glucose, Mn-NiO decorated carbon nanotubes, non-enzymatic

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Authors: Maha Ali Al-Mohaya, Lubna Majed Al-Otaibi, Ebtissam Nassir Al-Bakr, Abdulrahman Al-Asmari

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Objectives: Oral lichen planus (OLP) is a chronic inflammatory oral mucosal disease. Cytokines play an important role in the pathogenesis and disease progression of OLP. The purpose of this study was to investigate the association of tumor necrosis factor (TNF)-α, TNF-β and interleukin (IL)-10 gene polymorphisms with the OLP risk. Material and Methods: Forty-two unrelated patients with OLP and 211 healthy volunteers were genotyped for TNF-α (-308 G/A), TNF-β (+252A/G), IL-10 (-1082G/A), IL-10 (-819C/T), and IL-10 (-592C/A) polymorphisms. Results: The frequencies of allele A and genotype GA of TNF-α (-308G/A) were significantly higher while allele G and GG genotypes were lower in OLP patients as compared to the controls (P < 0.001). The frequency of GA genotype of TNF-β (+252A/G) was significantly higher in patients than in controls while the AA genotype was completely absent in OLP patients. These results indicated that allele A and genotype GA of TNF-α (-308G/A) as well as the GA genotype of TNF-β (+252A/G) polymorphisms are associated with OLP risk. The frequencies of alleles and genotypes of -1082G/A, -819C/T and -592C/A polymorphisms in IL-10 gene did not differ significantly between OLP patients and controls (P > 0.05). However, haplotype ATA extracted from 1082G/A, -819C/T, -592C/A polymorphisms of IL-10 were more prevalent in OLP patients when compared to controls indicating its possible association with OLP susceptibility. Conclusion: It is concluded that TNF-α (-308G/A), TNF-β (+252A/G) and IL-10 (-1082G/A, -819C/T and -592C/A) polymorphisms are associated with the susceptibility of OLP, thus giving additional support for the genetic basis of this disease. Further studies are required using a larger sample size to confirm this association and determine the prognostic values of these findings.

Keywords: oral lichen planus, cytokines, polymorphism, genetic

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Authors: Alaa M. M. Badr Eldin, Marwa I. Ezzat, Mohammed S. Sedeek, Manal S. Afifi, Omar M. Sabry

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Cytotoxic activity of the total methanol extract against breast cancer cell line MCF-7 was amazing IC50 at 54 ug/ml. 130 polyphenolic compounds were tentatively identified in pomegranate peel (Punica granatum L.) methanol extract using HPLC-MS/MS technique. The antiestrogenic activity of the polyphenolic constituents found in pomegranate extract was confirmed experimentally in-vitro and by the in-silico molecular docking using gallagic acid, ellagic acid, and Punicalagin as these are considered model compounds confirmed in pomegranate peel extract. The methanolic extract was found to suppress ER, TGF-β, and NF-kB in-vitro gene expression strongly, and that was verified by qPCR and Western Blot gel electrophoresis techniques.

Keywords: HPLC-MS/MS, pomegranate, breast cancer, ovarian cancer, ER, TGF-β, NF-kB

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Authors: Caio E. G. Reis, Sara Wassell, Adriana L. Porto, Angélica A. Amato, Leslie J. C. Bluck, Teresa H. M. da Costa

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Background: Multiple epidemiologic studies have consistently reported association between increased coffee consumption and a lowered risk of Type 2 Diabetes Mellitus. However, the mechanisms behind this finding have not been fully elucidated. Objective: We investigate the effect of coffee (caffeinated and decaffeinated) on glucose effectiveness and insulin sensitivity using the stable isotope minimal model protocol with oral glucose administration in healthy men. Design: Fifteen healthy men underwent 5 arms randomized crossover single-blinding (researchers) clinical trial. They consumed decaffeinated coffee, caffeinated coffee (with and without sugar), and controls – water (with and without sugar) followed 1 hour by an oral glucose tolerance test (75 g of available carbohydrate) with intravenous labeled dosing interpreted by the two compartment minimal model (225 minutes). One-way ANOVA with Bonferroni adjustment were used to compare the effects of the tested beverages on glucose metabolism parameters. Results: Decaffeinated coffee resulted in 29% and 85% higher insulin sensitivity compared with caffeinated coffee and water, respectively, and the caffeinated coffee showed 15% and 60% higher glucose effectiveness compared with decaffeinated coffee and water, respectively. However, these differences were not significant (p > 0.10). In overall analyze (0 – 225 min) there were no significant differences on glucose effectiveness, insulin sensitivity, and glucose and insulin area under the curve between the groups. The beneficial effects of coffee did not seem to act in the short-term (hours) on glucose metabolism parameters mainly on insulin sensitivity indices. The benefits of coffee consumption occur in the long-term (years) as has been shown in the reduction of Type 2 Diabetes Mellitus risk in epidemiological studies. The clinical relevance of the present findings is that there is no need to avoid coffee as the drink choice for healthy people. Conclusions: The findings of this study demonstrate that the consumption of caffeinated and decaffeinated coffee with or without sugar has no acute effects on glucose metabolism in healthy men. Further researches, including long-term interventional studies, are needed to fully elucidate the mechanisms behind the coffee effects on reduced risk for Type 2 Diabetes Mellitus.

Keywords: coffee, diabetes mellitus type 2, glucose, insulin

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Authors: Sara Mohammad Atef Sabaika

Abstract:

Background: The development of factor VIII (FVIII) inhibitor and hemophilic arthropathy in patients with hemophilia A (PWHA) are a great challenge for hemophilia care. Both genetic and environmental factors led to complications in PWHA. The development of inhibitory antibodies is usually induced by the immune response. Tumor necrosis factor α (TNF-α), one of the cytokines, might contribute to its polymorphism. Aim: Study the association between tumor necrosis alpha level and genotypes in pediatric patients with hemophilia A and its relation to inhibitor development and joint status. Methods: A cross-sectional study was conducted among a sufficient number of PWHA attending the Pediatric Hematology and Oncology Unit, Pediatric department in Menoufia University hospital. The clinical parameters, FVIII, FVIII inhibitor, and serum TNF-α level were assessed. The genotyping of −380G > A TNF-α gene polymorphism was performed using real time polymerase chain reaction. Results: Among the 50 PWHA, 28 (56%) were identified as severe PWHA. The FVIII inhibitor was identified in 6/28 (21.5%) of severe PWHA. There was a significant correlation between serum TNF-α level and the development of inhibitor (p = 0:043). There was significant correlation between polymorphisms of −380G > A TNF-α gene and hemophilic arthropathy development (p = 0:645). Conclusion: The prevalence of FVIII inhibitor in severe PWHA in Menoufia was 21.5%. The frequency of replacement therapy is a risk factor for inhibitor development. Serum TNF-α level and its gene polymorphism might be used to predict inhibitor development and joint status in pediatric patients with hemophilia A.

Keywords: hemophilic arthropathy, TNF alpha., patients witb hemophilia A PWHA, inhibitor

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Authors: Seyed Hassan Moosa-kazemi, Jalal Mohammadi Soleimani, Hassan Vatandoost, Mohammad Hassan Shirazi, Sara Hajikhani, Roonak Bakhtiari, Morteza Akbari, Siamak Hydarzadeh

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Malaria is an infectious disease and considered most important health problems in the southeast of Iran. Iran is elimination malaria phase and new tool need to vector control. Paraterasgenesis is a new way to cut of life cycle of the malaria parasite. In this study, the microflora of the surface and gut of various stages of Anopheles fluviatilis James as one of the important malaria vector was studied using biochemical and molecular techniques during 2013-2014. Twelve bacteria species were found including; Providencia rettgeri, Morganella morganii, Enterobacter aerogenes, Pseudomonas oryzihabitans, Citrobacter braakii، Citrobacter freundii، Aeromonas hydrophila، Klebsiella oxytoca, Citrobacter koseri, Serratia fonticola، Enterobacter sakazakii and Yersinia pseudotuberculosis. The species of Alcaligenes faecalis, Providencia vermicola and Enterobacter hormaechei were identified in various stages of the vector and confirmed by biochemical and molecular techniques. We found Providencia rettgeri proper candidate for paratransgenesis.

Keywords: Anopheles fluviatilis, bacteria, malaria, Paraterasgenesis, Southern Iran

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Authors: Liaquat Ali, Khurshid Natasha

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Low resource countries are not usually equipped with the organizational tools to implement health care for chronic diseases, and thus, providing effective diabetes care in such countries is a challenging task. Diabetic Association of Bangladesh (BADAS in Bengali acronym) has created a stimulating example to meet this challenge. Starting its journey in 1956 with 39 patients in a small tin shed clinic BADAS, and its affiliated associations now operate 90 hospitals and health centres all over the country. Together, these facilities provide integrated health care to about 1.5 million registered diabetic patients which constitute about 20% of the estimated diabetic population in the country. BADAS has also become a pioneer in health manpower generation in Bangladesh. Along with its affiliates, it now runs 3 Medical Colleges (to generate graduate physicians), 2 Nursing Institutes, and 2 Postgraduate Institutes which conduct 25 postgraduate courses (under the University of Dhaka) in various basic, clinical and public health disciplines. BADAS gives great emphasis on research, which encompasses basic, clinical as well as public health areas. BADAS is an ideal example of public-private partnership in health as most of its infrastructure has been created through government support but it is almost self-reliant in managing its revenue budget which approached approximately 40 million US dollar during 2010. BADAS raises resources by providing high-quality services to the people, both diabetic and non-diabetic. At the same time, BADAS has developed a cross financing model, to support diabetic patients in general and poor diabetic patients (identified through a social welfare network) in particular, through redistribution of the resources. Along with financial sustainability BADAS ensure organizational sustainability through a process of decentralization, community ownership, and democratic management. Presently a large scale pilot project (named as a Health Care Development Project or HCDP) is under implementation under BADAS umbrella with an objective to transform the diabetes care model to a health care model in general. It is expected to create further evidence on providing sustainable (with social safety net) health care delivery for diabetes, and other chronic illnesses as an integral part of general health care delivery in a resource constrained setting.

Keywords: Bangladesh, self sustain, health care, constrain

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Authors: Pratibha Srivastava, Ayyamperumal Jeyaprakash, Gary Steck, Jason Stanley, Leroy Whilby

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Exotic, invasive tephritid fruit flies (Diptera: Tephritidae) are a major threat to fruit and vegetable industries in the United States. The establishment of pest fruit fly in the agricultural industries and produce severe ecological and economic impacts on agricultural diversification and trade. Detection and identification of these agricultural pests in a timely manner will facilitate the possibility of eradication from newly invaded areas. Identification of larval stages to species level is difficult, but is required to determine pest loads and their pathways into the United States. The aim of this study is the New World genus, Anastrepha which includes pests of major economic importance. Mitochondrial cytochrome c oxidase I (COI) gene sequences were amplified from Anastrepha fraterculus specimens collected from South America (Ecuador and Peru). Phylogenetic analysis was performed to characterize the Anastrepha fraterculus complex at a molecular level. During phylogenetics analysis numerous nuclear mitochondrial pseudogenes (numts) were discovered in different specimens. The numts are nonfunctional copies of the mtDNA present in the nucleus and are easily coamplified with the mitochondrial COI gene copy by using conserved universal primers. This is problematic for DNA Barcoding, which attempts to characterize all living organisms by using the COI gene. This study is significant for national quarantine use, as morphological diagnostics to separate larvae of the various members remain poorly developed.

Keywords: tephritid, Anastrepha fraterculus, COI, numts

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Authors: Sheila M. Wicks, Gail Mahady, Udesh Patel, Joanna Michel, Armando Caceres

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Introduction: The genus piperaceae contains approximately 1000 species of herbs scrubs small trees and hanging vines distributed in both hemispheres. During our ethno medical work in Guatemala of the 27 plant families documented for us e by the Qeqchi Maya for reproductive disorders the most prominent were the Piperaceae (15%) and Menispermiaceae. Our Previous work showed that extracts from form Piper and Cissampelos species bound to both and progesterone and the estrogen receptors. In this work active extracts from Piper aeruginosibaccum Trelease, P auritum, P tuerckheimii and Cissampels tropaeolifolia were tested in functionalized cell based assays including a SEAP reporter gene and by qPCR of ER-responsive gene expression in MCF-7cells. In the reporter gene assay P aeruginosibaccum was estrogenic and enhanced E2 EFFECTS IN MCF-7 CELLS. P. tuerckheimi was not estrogenic alone but significantly enhanced the effects of E2 on SEAP reporter gene expression. Both altered mRNA expression of E2 responsive genes in MCF-7. Methods: this is collaborative project between University of Illinois at Chicago and University of San Carlos Guatemala City. 144 spices of plants were collected in Guatemala of which 57 used to treat a variety of women's reproductive health. The Genus Piperaraceae contains approximately 1000 species of herbs scrubs and small trees. Active extracts of the plants were tested in functionalized in cell-based bioassays including SEAP reporter genes. Results demonstrated altered mRNA expression of E2 responsive genes in MC-7 cells plants were collected in Guatemala of which 57 used. Conclusion of the 5 plants tested all were shown to contain components of binding to estrogenic receptor to a greater or lesser degree. These effects support the use of QEqchi Maya women in Guatemala for reproductive.

Keywords: reporter genes, MC7, guatemala piperaceae, reproductive health

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Authors: Ji Jiazheng, Wang Jingrao, Jin Xin, Zhang Hong

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Diabetes mellitus is a metabolic disease characterized by high blood sugar. A long-standing hyperglycemic state can lead to various tissue damage. Diabetic retinopathy is the most common and widely studied ocular complication and has become the leading cause of blindness in my country. At the same time, diabetes has profound clinically relevant effects on the cornea, leading to keratopathy and vision-threatening. The cornea is an avascular tissue and is sensitive to hyperglycemia, Keratopathy caused by diabetes is usually chronic, they are called diabetic keratopathy or diabetic neurotrophic keratopathy, leading to several diabetic corneal complications including delayed epithelial wound healing, recurrent erosions, neuropathy, loss of sensitivity. Corneal stem cell dysfunction in diabetic patients as an important influencing factor of diabetic keratopathy. The consequences of this condition are often underestimated. The limbus is located between the cornea and the sclera tissue. The limbal stroma consists of a series of radial elevations with fibrovascular centers known as palisades of Vogt (POV). Previous studies have shown that palisades of Vogt (POV), as the main site of limbal stem cells, plays an important role in the homeostasis of the corneal epithelium. Therefore, POV plays a vital role in the healing of corneal epithelial surgery and postoperative evaluation. IVCM can observe the condition of the corneal epithelium at the cellular level. It has profound significance and guidance for the evaluation of limbal and limbal stem cells. We have previously observed structural changes in POV in HSK and HZO patients on IVCM. At present, there have been reports involving limbal stem cell dysfunction in diabetic patients, but the specific pathogenesis is still unclear. However, there are no studies on POV morphological changes in patients with DM. Therefore, we performed statistics and compared the correlation between POV morphological changes and corneal epithelial basal cell density, corneal nerves, and length of disease in DM patients and normal humans using IVCM studies. At the same time, fundoscopy was used to observe the correlation between the thickness of RNFL and the thickness of GCC and POV in diabetic patients. And to observe the correlation between SVD, DVD and POV for research.

Keywords: confocal microscopy, fundus, limbal stem cells, diabetes

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