Potential pitfalls in the accuracy of analysis of natural sense-antisense RNA pairs by reverse transcription-PCR

<p>Abstract</p> <p>Background</p> <p>The ability to accurately measure patterns of gene expression is essential in studying gene function. The reverse transcription polymerase chain reaction (RT-PCR) has become the method of choice for the detection and measurement of RNA expression patterns in both cells and small quantities of tissue. Our previous results show that there is a significant production of primer-independent cDNA synthesis using a popular RNase H^-RT enzyme. A PCR product was amplified from RT reactions that were carried out without addition of RT-primer. This finding jeopardizes the accuracy of RT-PCR when analyzing RNA that is expressed in both orientations. Current literature findings suggest that naturally occurring antisense expression is widespread in the mammalian transcriptome and consists of both coding and non-coding regulatory RNA. The primary purpose of this present study was to investigate the occurrence of primer-independent cDNA synthesis and how it may influence the accuracy of detection of sense-antisense RNA pairs.</p> <p>Results</p> <p>Our findings on cellular RNA and <it>in vitro </it>synthesized RNA suggest that these products are likely the results of RNA self-priming to generate random cDNA products, which contributes to the loss of strand specificity. The use of RNase H⁺RT enzyme and carrying the RT reaction at high temperature (50°C) greatly improved the strand specificity of the RT-PCR detection.</p> <p>Conclusion</p> <p>While RT PCR is a basic method used for the detection and quantification of RNA expression in cells, primer-independent cDNA synthesis can interfere with RT specificity, and may lead to misinterpretation of the results, especially when both sense and antisense RNA are expressed. For accurate interpretation of the results, it is essential to carry out the appropriate negative controls.</p

A Retrospective Appraisal of Teacher Induction

A Retrospective Appraisal of Teacher Induction Abstract Examination of an induction program for new teachers was undertaken from the viewpoint of induction graduates three years after participation. Their retrospective perspectives were investigated as to their satisfaction with assimilation in school in the induction year, their attitudes towards organizational aspects of the program, and the program\u27s contribution to their professional development. Comparisons were made to beginning teachers in the midst of their induction year. Data were collected from 98 induction graduates and 390 induction participants using questionnaires. Compared to induction participants, graduates retrospectively remembered the induction year at school less positively and more often recommended extending induction support. The graduates ascribed only moderate contribution to the induction program. In general retrospective appraisals of active teachers and non-teaching graduates were similar. Implications for the use of retrospective evaluations are discussed

Predictors of Teacher Educators\u27 Research Productivity

This study examined the relationship between teacher educators\u27 research productivity (RP) and their background and professional characteristics, attitudes, motives, obstacles and time devoted to research. The sample included 161 teacher educators from four teacher education colleges in Israel. The findings indicate the significance of five variables for predicting RP: academic degree, rank, administrative position, desire to develop new knowledge and learn from research findings and perceived insufficient research competence and self-confidence. These variables account for 37.2% of the variance in RP. The results from this study provide useful information for teacher education institutions and policy makers regarding variables significantly related to RP. These variables should be addressed when recruiting teacher educators, assigning administrative duties and designing professional development programs, particularly for new career faculty

The Effect of Corporate Cultural Factors on the Data Warehousing Success: An Empirical Investigation

This study is aimed at identifying the impact of corporate cultural factors on successful development of data warehousing in the United Arab Emirates. The theoretical framework of the study is formulated based on analysis of related literature coupled with the information gained from interviewing data warehousing experts. Five hundred and eighty data warehouse users in 34 companies were surveyed to obtain their perceptions of the extent that each of 132 items had actually contributed to their firms’ DW success at different phases of development. Rigorous multivariate statistical analysis procedure has been followed to design and construct an overall model of DW success. The model has proven that all its independent variables have significant influence on the DW overall success and that corporate cultural factors have dominant impact on this success throughout the different phases of DW development

Economic implications of comorbid conditions among Medicaid beneficiaries with COPD

SummaryObjectivesTo characterize a comprehensive comorbidity profile and to explore the economic implications of comorbidity among patients with chronic obstructive pulmonary disease (COPD).MethodsThis retrospective cohort study analyzed medical claims from the Maryland Medicaid database. We employed a 1:2 case–control design to select COPD patients (n=1388) and demographically matched controls (n=2776) aged 40 to 64 years with 24 months of continuous enrollment. Odds ratios were employed to compare comorbidity differences, including 17 conditions defined by the Charlson Comorbidity Index (CCI) and 6 additional conditions commonly observed in COPD patients. We estimated the incremental medical utilization and medical cost by specific condition.ResultsCompared with the controls, Medicaid COPD patients had higher comorbidity burden and were more likely to have myocardial infarction, congestive heart failure, cerebrovascular disease, peptic ulcer, mild liver disease, hypertension, sleep apnea, tobacco use, and edema. COPD patients on average had 24% more medical claims (81.4 vs. 65.4, p<0.001) and were 33% more expensive than controls ($7603 vs.$5732, p<0.001). Ten conditions defined by the CCI as well as hypertension, tobacco use, and edema were associated with incremental medical utilization and cost in COPD patients; depression was associated with incremental medical utilization but not cost.ConclusionsThe high burden of comorbidity in COPD patients translates into additional medical utilization and cost. Effective disease management and treatment protocols are needed to reduce comorbidity burden. The development of a COPD-specific comorbidity measure may be used to identify high-risk subgroups and to predict utilization and cost

Influence of Polyethylene Soil Mulch and Foliar Application of Urea , Complete Fertilizers, Seaweed in Alleviating Salt Stress of Broccoli (Brassica Oleracea Var.Italica)

This experiment was carried out to study the effect of soil mulch (with black polyethylene) and foliar application of  urea, complete fertilizer and seaweed as well as control) and their interaction in alleviation of salt stress on broccoli which cultivated in salty soil(11.3 dS.m-1) under drip irrigation , by estimating of leaf area, SOD and Catalase activity , MDA and Glutathione concentration in both of leaves and flowers.  In broccoli leaves, soil mulch gave a significant effect in each of leaf area, SOD activity and non- significant effect in catalase activity, Glutathione  and MDA concentration , while in flowers it caused a significant effects on all studied parameters except catalase activity compared to no mulch treatment . Complete fertilizer treatment caused higher effect in all studied parameters than urea and seaweed compared to control treatment . In all studied parameters , the interaction between soil mulch and foliar treatments had high significant effect . complete fertilizer + mulch treatment was the  best, which caused increase in leaf area , SOD and Catalase activity in addition of Glutathione concentration compared to control treatment in both leaves and flowers , while it caused the biggest significant decrease in MDA concentration in both of leaves and flowers. Keywords: Broccoli, Foliar fertilizer, seaweed , soil mulch, Urea, complete fertilize

Application of Sharia Principles in Sharia Financial Institutions

This study analyzes related to Islamic Financial Institutions (LKS). Which currently exists and is growing quite rapidly. There have been numerous variations of LKS throughout Indonesia, including Islamic institutions. LKS is a financial institution that operates following Sharia law. LKS, a financial institution with Sharia principles, was originally present as an option and a solution for Muslims who want to avoid the practice of conventional banks or financial institutions. LKS was originally offered as a sharia-compliant financial organization for Muslims who want to escape the practice of banks or traditional financial institutions that use the interest system, but they can also be a choice for non-Muslims. Therefore, there are problems why the application of Sharia principles by LKS is an urgent problem and what are the conditions for applying Sharia principles by LKS. The research method used is normative legal research with a case approach. The study results indicate that the LKS organizer is expected to have a vision in which the LKS remains in the principles of Sharia

Evaluated the Up –regulation in Gene ‎Expression of Hepatic Insulin Gene and ‎Hepatic Insulin Receptor Gene in Type 1 ‎Diabetic Rats Treated with Cuscuta chinesis ‎Lam.‎

&nbsp; &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;انجزت هذه الدراسة للكشف عن فعالية مستخلص نبات الحامول الخافض للسكر &nbsp;ودراسة ألية عمله على المستوى الجزيئي &nbsp;والنسيجي &nbsp;للاعضاء ذات العلاقة بمرض السكر( البنكرياس والكبد)، حيث تم قياس &nbsp;العديد&nbsp; من المؤشرات مثل مستوى سكر الدم الصيامى، مستوى الانسولين الصيامى في المصل، التعبير الجيني لكل من جين الانسولين في الكبد، جين الانسولين في البنكرياس و جين مستقبل الانسولين في الكبد بالاضافة الى اجراء فحوصات لمقاطع في انسجة البنكرياس والكبد. لاجراء هذه التجربة تم تقسيم اربعا وخمسون ذكراً من الجرذا ن نوع ٌRattus&nbsp; rattus&nbsp; والتي تراوحت معدلات&nbsp; أوزنها من&nbsp; 200-180 غم&nbsp; الى ثلاثة مجاميع رئيسية: مجموعة أ (مجموعة السيطرة للجرذان الطبيعي والتي تم تجريعها بالماء المقطر يومياً)، مجموعة ب(مجموعة السيطرة للجرذان المصابة&nbsp; بداء السكري النوع الاول والتي تم تجريعها ايضاً بالماء المقطر يومياً) اما المجموعة الثالثة ج فكانت (مجموعة الجرذان المصابة&nbsp; بداء السكري النوع الاول والتي تم تجريعها يومياً بالمستخلص الميثانولي بمقدار 400 ملغم/ كغم من وزن الجسم). تم تقسم كل مجموعة رئيسية والتي كانت تتالف من 18 جرذا الى ثلاث مجاميع ثانوية وفقا لمدة التجريع : مجوعة رقم (1) ، مجموعة رقم 2 ومجموعة رقم 3 لتشير الى عدد أيام التجريع 30 ، 60 و 90 يوما على التوالي ولقد تم قياس جميع المؤشرات السابقة وأوضحت النتائج بان التجريع اليومي بمستخلص نبات الحامول &nbsp;400ملغم/كغم وزن جسم ادى الى الانخفاض المعنوي لمستوى سكر الدم الصيامي للمجموعة ج قياسا &nbsp;بالمجموعة ب وكذلك رفع من مستوى كل من الانسولين الصيامى في المصل و، مستوى التعبير الجيني لكل من جين الانسولين في الكبد ، جين الانسولين في البنكرياس و جين مستقبل الانسولين في الكبد في المجموعة ج قياسا بالمجموعة ب ، كما ونلاحظ بانه اعاد جميع المؤشرات المدروسة في المجموعة ج الى الحالة الطبيعية بضمنها الطبيعة النسيجية لكل من النكرياس والكبد. استنتجت هذه الدراسة بان التجريع اليومي للجرذان المصابة بداء السكري من النوع الاول ب400ملغم ملغم/كغم من وزن الجسم من مستخلص نبات الحامول ولمدة 60 يوماً أعاد مستوى سكر الدم الصيامي الى مستواه الطبيعي من خلال رفع مستوى التعبير الجيني لكل من جين الانسولين في الكبد، جين الانسولين في البنكرياس&nbsp; ،جين مستقبل الانسولين في الكبد ، اضافة الى ترميم خلايا بيتا في البنكرياس واعادة التركيب النسيجي لكل من البنكرياس والكبد الى الحالة الطبيعية&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; This research was conducted to study the hypoglycemic activity of C. chinesis Lam on type 1 diabetic disease and investigate the &nbsp;molecular and histological mechanism of &nbsp;its action .many parameters was investigated , Fasting blood glucose (FBG), Fasting serum insulin,Hepatic Insulin Gene Expression, pancreas Insulin Gene Expression ,Hepatic Insulin&nbsp; Receptors Gene expression &nbsp;and histological sections of pancrease and liver.54 Rattus rattus male rats weighting(180 -200g) were divided into 3 groups: A normal control daily administrated with Dw, B Diabetic control daily administrated with Dw&nbsp; and C&nbsp; diabetic group daily administrated with 400 mg/Kg body weight of C. chinesis&nbsp; Lam. methanolic extract, each group consisted of&nbsp; 18 rats and further divided into (3) sub- groups 1 ,2 &nbsp;and 3. According to the period of administration&nbsp; 30, 60 and&nbsp; 90 days respectively. The results showing &nbsp;the daily administration of 400 mg/Kg body weight of C. chinesis&nbsp; Lam. methanolic extract for 60 day causing significance&nbsp; decrease&nbsp; in FBG and In the other hand each of fasting serum insulin, hepatic Insulin gene expression,pancreas Insulin gene expression and hepatic Insulin receptor gene expression was increased in group C in compare to B group and return all studied parameters involving pancrease and liver texture to the normal state ,which were statically morphologically &nbsp;not appeared any significant difference from A group .this study concluded that the daily administration type 1 diabetic rats with 400 mg/Kg body weight of C. chinesis&nbsp; Lam. extract for 60 day was return&nbsp; fasting serum insulin and FBG to normal value by &nbsp;upregulated&nbsp; the gene expression of hepatic INS Gene ,INSR gene , pancreas INS Gene ,regenerate pancreatic beta- cell and returnthe texture of both liver and pancrease to the normal stat