Stable suppression of gene expression by short interfering RNAs targeted to promoter in a mouse embryonal carcinoma stem cell line

RNA interference (RNAi) can induce gene silencing via two pathways: post-transcriptional gene silencing (PTGS) and transcriptional gene silencing (TGS). The mediators of gene inactivation in both pathways are 21-bp small interfering RNAs (siRNAs) generated from longer double-stranded RNA (dsRNA). PTGS involves siRNA-mediated targeting and degradation of mRNA. However, siRNAs induce TGS via DNA methylation at the targeted promoter. Synthetic siRNAs can induce loss of gene activity comparable to long dsRNA. The limitation of this method is that the transfected synthetic siRNA works for only a few days. In this study, we tested the RNAi response to siRNA (PTGS pathway) by using a plasmid containing an enhanced green fluorescent protein (eGFP) gene as a target as well as a plasmid creates siRNA transcript, in a form of a hairpin, against eGFP gene. To investigate TGS pathway via RNAi, we also used a plasmid creates hairpin siRNA transcript against pgk-1 promoter. The data presented here indicated long-lasting inhibition in expression of eGFP and puromycin genes, both under the control of the murine Pgk-1 promoter. However, Southern blot analysis showed no methylation in pgk-1 promoter. © 2010 The Society for In Vitro Biology

Neuronal differentiation of GFP expressing P19 embryonal carcinoma cells by deprenyl, an antiparkinson drug

زمینه و هدف: سلول های P19 دودمانی از سلول های کار سینومای جنینی چند استعدادی هستند که قادرند در محیط کشت حاوی سرم رشد نموده و برای تمایز به هر سه دودمان مزودرمی، آندودرمی و اکتودرمی القا شوند. با استفاده از روش های استاندارد ترانسفکشن می توان توالی DNA خارجی را به این سلول ها وارد کرده و عملکرد و تمایز سلولی را بررسی کرد. این تحقیق با هدف القای فنوتیپ عصبی در سلول های P19 ترانسفکت شده با ژن پروتئین فلوئورسانت سبز (GFP) با استفاده از داروی ضد پارکینسونی دپرنیل صورت گرفت. روش بررسی: در این مطالعه توصیفی آزمایشگاهی از روش رسوب کلسیم فسفات جهت وارد کردن پلاسمید pML8 حاوی ژن GFP و ژن مقاوم به پیورومایسین به این سلول ها استفاده شد. سلول ها با استفاده از محیط کشت MEM-A (Minimum Essential Medium Alpha) حاوی 15 درصد سرم گاوی جنینی کشت داده شدند. در اینجا غلظت 8-10 مولار دپرنیل برای القای تمایز اجسام شبه جنینی به دودمان عصبی استفاده شد. جهت ارزیابی تمایز سلول های P19 بررسی موفولوژی با استفاده از رنگ آمیزی اختصاصی کرزیل ویوله انجام شد. به علاوه برای ردیابی پروتئین های ویژه سلول های عصبی مانند سیناپتوفیزین و بتاتوبولین III از روش ایمنوفلورسنس استفاده شد. یافته ها: ضمن تولید سلول های P19 ترانسفکت شده پایدار، تمایز عصبی این سلول ها تحت تأثیر عامل القایی دپرنیل انجام شد. نورون های GFP مثبت مشتق شده از سلول های کار سینومای جنینی نشانگرهای ویژه سلول های عصبی را بیان کردند. نتیجه گیری: سلول های تمایز یافته GFP مثبت می توانند در مطالعات پیوندی و پژوهش های پایه ای مرتبط با سلول درمانی و بیماری های تحلیل سیستم عصبی مورد استفاده قرار گیرند

Endometrial and follicular development following stair-step and traditional protocols in women with polycystic ovary syndrome: An RCT

Background: Various strategies have been proposed for polycystic ovary syndrome (PCOS) treatment. Objective: To investigate and compare the number and size of ovarian follicles, endometrial thickness, and ovulation rate by traditional protocol (TP) and stair-step protocol (SSP). Materials and Methods: Sixty infertile PCOS women were allocated into two groups (SSP = 30 and control TP = 30) between May and October 2019 in the Besat Hospital, Sanandaj, Iran. In the SSP group, the infertile women were treated with 50 mg/daily clomiphene citrate (CC) for five days, while the nonresponsive women were prescribed 100 mg daily CC for five days in the same cycle. The maximum dose (150 mg) was administered until ovulation occurred. In the control group, in non-ovulatory cases, the dose was increased in the next cycle. Ultrasound was used to detect ovulation. Results: Endometrial thickness changes with various doses of CC were significantly different in the TP. The comparison of both protocols showed a significant difference in endometrial thickness only at 50 mg CC. The number of follicles in the left ovary was significantly different in both protocols at 150-mg CC. The size of ovarian follicles in the left ovary was significantly different between the two protocols at 100-mg CC. The ovulation rate was significantly different in the SSP at 100- and 150-mg doses of CC. Moreover, 86% of ovulation occurred at 100-mg CC in the SSP, while this rate was 73% in the TP. Conclusion: The most appropriate dose for ovulation in patients with PCOS is 100 mg CC. Key words: Polycystic ovary syndrome, Clomiphene, Infertility, Ovulation induction

The Correlation between Iranian EFL Learners’ Intercultural Sensitivity, Vocabulary Knowledge, and English Language Proficiency

AbstractThis study aimed to explore any type and level of association between Iranian EFL learners’ proficiency level and their intercultural sensitivity on the one hand, and the possible relation between vocabulary knowledge and sensitivity to cultural differences on the other. To this end, Oxford Quick Placement Test (OQPT) was administered to 220 EFL leaners. Based on the results of this test, a homogenized sample of 150 EFL learners (70 male and 80 female) at intermediate and upper-intermediate levels was selected. Afterwards, the participants took Vocabulary Levels Test (VLT) developed by Nation, and validated by Webb, Sasao, and Balance. Finally, Intercultural Sensitivity Scale (ISS) by Chen & Starosta was administered. The results of Pearson correlation analyses revealed a statistically significant correlation between the participants' proficiency level and their intercultural sensitivity. The results of regression analyses also indicated that language proficiency contributes as much as 55.4 percent to the prediction of level of intercultural sensitivity. Moreover, a significant positive correlation was detected between EFL learners’ L2 vocabulary knowledge and their intercultural sensitivity level. Further, the results revealed that Iranian EFL learners’ L2 vocabulary knowledge can offer contributions up to 17.3 percent to the prediction of intercultural sensitivity level. These findings can offer prominent implications for all practitioners, material developers, and EFL instructors who are primarily preoccupied with linguistic competence. The results can motivate them to consider intercultural sensitivity as a complementary element to EFL learners’ linguistic knowledge, as well as their communicative commands

Investigating the Mediating Role of Organizational Commitment in the Relationship Between Mentoring and Job Satisfaction

Extended AbstractAbstract The aim of the present study is to investigate the mediating role of organizational commitment in the relationship between mentoring and job satisfaction and to answer the question of whether organizational commitment plays a mediating role in the relationship between mentoring and job satisfaction  approach: In terms of purpose, this research is applied, and according to the nature of the subject and its goals, it is of a descriptive-correlation type.Introduction Education and its scientific and specialized management is an important factor in the development of countries and societies. Today, coaching is considered the most important experiential aspect of training and is an important process for professional development and psychological support of new employees, and it can be used to prevent anxiety and increase self-confidence and peace of mind in inexperienced employees.Case study The statistical population of the present study was all primary school teachers in Euclid city. The required statistical sample size was determined by Cochran's formula of 134 people and the sample was selected using random sampling method. The required primary data were collected through questionnaires and secondary data were collected through library and internet sources.Materials and Methods Structural equation model was used by Smart PLS software to test research hypotheses. The required statistical sample size was determined by Cochran's formula of 134 people and the sample was selected using random sampling method. The required primary data were collected through questionnaires and secondary data were collected through library and internet sources.Discussion and Results The research findings showed that in general, the mentoring variable explains a total of 59% of the variance of the job satisfaction variable and 19.9% of the variance of the organizational commitment variable. And the variable of organizational commitment explains a total of 53% of the variance of the variable of job satisfaction, according to the mentioned cases, all research hypotheses were confirmed. The lack of easy access to teachers and members of the statistical community as well as the lack of research literature are among the limitations of the research.Conclusionpresent research include the achievement that by combining the social exchange theory (which is one of the most effective conceptual paradigms for understanding workplace behaviors) and the concept of coaching, it is possible to develop a suitable structure for predicting and explaining the behavioral dimensions between the organization and employees. Finally, one of the acquired concepts is training methods of persuading employees by managers and supervisors in order to achieve commitment, job integration, more self-efficacy and less intention to leave the organization

Antidepressant-like effect of essential oil of Lavandula angustifolia Mill and Citrus bigaradia Duh with forced swimming test in reserpinized mice Balb/C.

Background: Levandula angustifolia and Citrus aurantium essential oils possess phenolic compounds and in Iranian traditional medicine are commonly used as sedative. The aim of this study was to evaluate the antidepressant activities of Levandula angustifolia and Citrus bigaradia in adult reserpinized mice. Methods: In this experimental study, mice were divided into 10 groups of 8 mice each, including: Group 1, control group received normal saline. Group 2, was treated reserpine (5 mg/kg, IP). Group 3, 4, 5 and 6 were treated with reserpine (5 mg/kg, IP) and four Citrus aurantium different doses (25, 50, 100, 200 mg/kg). Group 7, 8 and 9 were treated with reserpine (5 mg/kg, IP) and three Levandula angustifolia doses (25, 50, 100 mg/kg). Group 10 was treated with reserpine (5 mg/kg, IP) and imipramine (15 mg/kg). In all groups the forced swimming test was done and the level of malondialdehyde (MDA) in brain tissue was measured to estimate the oxidative stress status. Data were analyzed by one-way ANOVA and Tukey post hoc test. Results: Levandula angustifolia, Citrus aurantium and imipramine in reserpinized mice caused the mice to spend less time in the forced swimming test, compared to the control group (p<0.01). Also Citrus aurantium with dose of 50 and 100 mg/kg and Lavandula angustifolia with dose of 25 and 50 mg/kg in reserpinized mice reduced immobility time and levels of MDA compared the control group (p<0.01). Conclusion: Lavander and Citrus aurantium essential oils have considerable antidepressant activity which might be attributed to their antioxidant effects

Investigating the Relationship Between High Levels of Angiotensin II in Plasma and the Rate of Deterioration in Hypertensive Patients to COVID-19

Introduction: The COVID-19 crisis poses a heightened risk to vulnerable populations. Considering factors influencing deterioration, complications, and mortality, it is especially important to pay attention to these groups, including persons with hypertension (HTN) and diabetes. This study aimed to investigate the correlation between the angiotensin II (ANG II) level and the disease severity and clinical course in COVID-19 patients. Methods: A cross-sectional study was conducted on 50 COVID-19 patients (mean age 59.1±20) admitted to Sinai hospital in Hamedan. The blood samples of the patients were taken during hospitalization and discharge, and the plasma ANG II level was measured using an enzyme-linked immunosorbent assay kit. The quantitative comparison was analyzed with paired t test and qualitative comparison with the chi-square test. The correlation of variables was checked with the Pearson test, and P<0.05 was considered statistically significant. Results: Overall, 44%, 16%, and 10% of COVID-19 patients had HTN, cardiovascular disease (CVD), and 10% diabetes, respectively. The oxygen level of 82% of the patients was below 90, of which 68% were intubated, and the lowest oxygen levels were observed in patients with HTN and CVD with 2 deaths. The primary and secondary examination of the ANG II level demonstrated that its level was significantly higher during illness compared to full recovery and decreased during recovery. Conclusion: The ANG II level is related to the severity of the disease in the early stages of 2019- nCoV infection. Therefore, HTN or other diseases that affect the level of ANG II in the blood can increase the severity of the disease

First evidence of the presence of adenovirus type 8 in myocardium of patients with severe idiopathic dilated cardiomyopathy

Previous studies have detected adenovirus and cytomegalovirus (CMV) in cardiac tissue of patients with myocarditis. Therefore, in this study, we investigated the frequency of these viruses, which may be involved in the development of severe dilated cardiomyopathy (DCM). Myocardial tissue from of 23 cardiac transplant candidates with acute idiopathic DCM below the age of 40 years were analyzed by amplification of adenovirus and CMV DNA and subsequent sequencing. Adenovirus was detected in four (17.4%) and CMV in one (4.3%) of the patients. All controls were negative for the presence of both viruses. Our study shows that myocardial infection with adenovirus may play an important role in the pathogenesis of severe DCM and suggests that vaccination against adenovirus might be helpful in decreasing the prevalence of severe idiopathic DCM. This is the first study in which adenovirus type 8 has been detected in the hearts of patients with DCM

The unfinished agenda of communicable diseases among children and adolescents before the COVID-19 pandemic, 1990-2019: a systematic analysis of the Global Burden of Disease Study 2019

BACKGROUND: Communicable disease control has long been a focus of global health policy. There have been substantial reductions in the burden and mortality of communicable diseases among children younger than 5 years, but we know less about this burden in older children and adolescents, and it is unclear whether current programmes and policies remain aligned with targets for intervention. This knowledge is especially important for policy and programmes in the context of the COVID-19 pandemic. We aimed to use the Global Burden of Disease (GBD) Study 2019 to systematically characterise the burden of communicable diseases across childhood and adolescence. METHODS: In this systematic analysis of the GBD study from 1990 to 2019, all communicable diseases and their manifestations as modelled within GBD 2019 were included, categorised as 16 subgroups of common diseases or presentations. Data were reported for absolute count, prevalence, and incidence across measures of cause-specific mortality (deaths and years of life lost), disability (years lived with disability [YLDs]), and disease burden (disability-adjusted life-years [DALYs]) for children and adolescents aged 0-24 years. Data were reported across the Socio-demographic Index (SDI) and across time (1990-2019), and for 204 countries and territories. For HIV, we reported the mortality-to-incidence ratio (MIR) as a measure of health system performance. FINDINGS: In 2019, there were 3·0 million deaths and 30·0 million years of healthy life lost to disability (as measured by YLDs), corresponding to 288·4 million DALYs from communicable diseases among children and adolescents globally (57·3% of total communicable disease burden across all ages). Over time, there has been a shift in communicable disease burden from young children to older children and adolescents (largely driven by the considerable reductions in children younger than 5 years and slower progress elsewhere), although children younger than 5 years still accounted for most of the communicable disease burden in 2019. Disease burden and mortality were predominantly in low-SDI settings, with high and high-middle SDI settings also having an appreciable burden of communicable disease morbidity (4·0 million YLDs in 2019 alone). Three cause groups (enteric infections, lower-respiratory-tract infections, and malaria) accounted for 59·8% of the global communicable disease burden in children and adolescents, with tuberculosis and HIV both emerging as important causes during adolescence. HIV was the only cause for which disease burden increased over time, particularly in children and adolescents older than 5 years, and especially in females. Excess MIRs for HIV were observed for males aged 15-19 years in low-SDI settings. INTERPRETATION: Our analysis supports continued policy focus on enteric infections and lower-respiratory-tract infections, with orientation to children younger than 5 years in settings of low socioeconomic development. However, efforts should also be targeted to other conditions, particularly HIV, given its increased burden in older children and adolescents. Older children and adolescents also experience a large burden of communicable disease, further highlighting the need for efforts to extend beyond the first 5 years of life. Our analysis also identified substantial morbidity caused by communicable diseases affecting child and adolescent health across the world. FUNDING: The Australian National Health and Medical Research Council Centre for Research Excellence for Driving Investment in Global Adolescent Health and the Bill & Melinda Gates Foundation

Designing and establishment of ISO/IEC 17025 in laboratories of national inland water aquaculture center and south Iran aquaculture research center

The project was carried out between June of 2011 and November of 2012,8 laboratories of research center in Anzali (Plankton, Algae, Hydrochemistry, Physiology, Ichthyology, Bentose, Parazitology, Virology) and 7 laboratories of research center in Ahvaz (Clinical pathology, Plankton, Hydrochemistry, Physiology, Ichthyology, Bentose, Parazitology, Virology) were selected for accreditation. The main stages for establishment of the system consisted of: 1-Conducting a gap analysis to compare the present state of the laboratories with ISO/IEC 17025 2-Training General requirements for the competence of testing and calibration laboratories Validation of methods Estimation of uncertainty Internal audits 3- Performing of technical and management requirements 4-Submit of quality manual to ASCB center in England in order to accredit In August of 2012 The main results were including: 1-Increase the accuracy of measurement in laboratories 2-Improvement of the Repeatability and Reproducibility of the test methods 3-Traceability and standardization of test methods 4- Calibration of measurement instruments 6- Updating of test methods 7-Standardization of physical condition of the laboratories 8- Getting the certification from ASCB center i