Altered expression of cell cycle and apoptotic proteins in chronic hepatitis C virus infection.

RIGHTS : This article is licensed under the BioMed Central licence at http://www.biomedcentral.com/about/license which is similar to the 'Creative Commons Attribution Licence'. In brief you may : copy, distribute, and display the work; make derivative works; or make commercial use of the work - under the following conditions: the original author must be given credit; for any reuse or distribution, it must be made clear to others what the license terms of this work are.BACKGROUND: A disrupted cell cycle progression of hepatocytes was reported in chronic hepatitis C virus (HCV) infection, which can contribute significantly in the associated pathogenesis. The present study aimed to further elaborate these disruptions by evaluating the expression of key cell cycle and apoptotic proteins in chronic HCV infection with particular reference to genotype 3. Archival liver biopsy specimens of chronic HCV-infection (n = 46) and normal histology (n = 5) were analyzed by immunohistochemistry using antibodies against proliferation marker Mcm-2, G1 phase marker Cyclin D1, S phase marker Cyclin A, cell cycle regulators p21 (CDK inhibitor) and p53 (tumor suppressor protein), apoptotic protein Caspase-3 and anti-apoptotic protein Bcl-2. RESULTS: Elevated Mcm-2 expression was observed in hepatocytes in chronic HCV infection, indicating increased cell cycle entry. Cyclin D1 expression was higher than cyclin A, which suggests a slow progression through the G1 phase. Expression of cell cycle regulators p21 and p53 was elevated, with no concordance between their expressions. The Mcm-2 and p21 expressions were associated with the fibrosis stage (p = 0.0001 and 0.001 respectively) and that of p53 with the inflammation grade (p = 0.051). Apoptotic marker, Caspase-3, was mostly confined to sinusoidal lining cells with little expression in hepatocytes. Anti-apoptotic protein, Bcl-2, was negligible in hepatocytes and detected principally in infiltrating lymphocytes. Expression of all these proteins was unrelated to the HCV genotype and were detected only rarely in the hepatocytes of normal liver. CONCLUSION: The results showed an arrested cell cycle state in the hepatocytes of chronic HCV infection, regardless of any association with genotype 3. Cell cycle arrest is characterized by an increased expression of p21, in relation to fibrosis, and of p53 in relation to inflammation. Furthermore, expression of p21 was independent of the p53 expression and coincided with the reduced expression of apoptotic protein Caspase-3 in hepatocytes. The altered expression of these cell cycle proteins in hepatocytes is suggestive of an impaired cell cycle progression that could limit the regenerative response of the liver to ongoing injury, leading to the progression of disease

Rewards Satisfaction, Perception about Social Status and Commitment of Nurses in Pakistan

This research is undertaken to approve or reject the general perception of masses about commitment of employees and to identify those factors, which effect commitment of employees in general and of nurses in particular. In previous research of Allen and Myere (1990), the factor of social status was not addressed. In this study target audience is nurses. Whereas, it is a proven fact that psychological perception effects individual’s attitudes and behaviors, so to see that how satisfaction with rewards/ benefits and perception about social status effect commitment of employees, this research was undertaken. Satisfaction level varies from person to person. The empirical results have proved that commitment of employees is affected due to satisfaction with rewards/ benefits and perception about social status

Rewards Satisfaction, Perception about Social Status and Commitment of Nurses in Pakistan

This research is undertaken to approve or reject the general perception of masses about commitment of employees and to identify those factors, which effect commitment of employees in general and of nurses in particular. In previous research of Allen and Myere (1990), the factor of social status was not addressed. In this study target audience is nurses. Whereas, it is a proven fact that psychological perception effects individual’s attitudes and behaviors, so to see that how satisfaction with rewards/ benefits and perception about social status effect commitment of employees, this research was undertaken. Satisfaction level varies from person to person. The empirical results have proved that commitment of employees is affected due to satisfaction with rewards/ benefits and perception about social status

Fekete-Szegö Problem of Functions Associated with Hyperbolic Domains

In the field of Geometric Function Theory, one can not deny the importance of analytic and univalent functions. The characteristics of these functions including their taylor series expansion, their coefficients in these representations as well as their associated functional inequalities have always attracted the researchers. In particular, Fekete-Szegö inequality is one of such vastly studied and investigated functional inequality. Our main focus in this article is to investigate the Fekete-Szegö functional for the class of analytic functions associated with hyperbolic regions. Tofurther enhance the worth of our work, we include similar problems for the inverse functions of these discussed analytic functions

Assessment of knowledge, preventive behaviours and risk perception related to Covid-19 among medical students of Rawalpindi

Objectives: The study was conducted to determine the knowledge, preventive behaviours and perception of risk related to Covid-19 among the medical students who are the front line workers and are more susceptible to be infected. Methodology: This analytical cross sectional study was conducted in medical students of 1st, 2nd and 3rd year MBBS of various medical institutions of Rawalpindi. Study duration was 5 months (March 2020 to July 2020). The sample size was 425. Age ranging from 18 to 21 and 273 (64.2%) comprised of females. The inclusion criteria were those students who gave consent and filled out the questionnaire and the exclusion criteria was who didn't concede. The data was analysed on SPSS. Confidence interval was set to be 95% with 5% margin of error. Results: Most of the students were aware of the basic Covid-19 related knowledge like common symptoms, incubation period, transmission and the basic preventive measures which was on average higher than the previous studies conducted on medical students and on health care workers. Students were also having high level of performance in preventive behaviours in perspective of Covid-19 but risk perception is of moderate level. Conclusion: Participants were found to have high levels of knowledge related to Covid-19 except use of N95 masks and use of anti-viral drugs for treatment. Among self-reported preventive behaviours section, lowest score is related to frequency of cleaning and disinfecting items that can be easily touched with hands (i.e. door handles and surfaces). Participants had moderate risk perception despite having high levels of knowledge and high performance in self-reported behaviours. Keywords: Covid-19, medical students, risk perceptio

Lessons Drawn From Pakistan-Sri Lanka FTA

There has been an increase in bilateral trade between Sri Lanka and Pakistan in the post – FTA milieu. Today Pakistan is the second largest trading partner of Sri Lanka amongst South Asian economies. There have been some concerns about the negative trade balance for Sri Lanka however the lower prices have resulted in an increase in welfare for both countries. The negative trade balance for Sri Lanka can also be justified on the account that a significant proportion of Sri Lankan imports from Pakistan constitute raw material and related intermediate inputs that in turn lead to efficiency gains. Most noticeable are the over 30 percent imports from Pakistan, currently being used as raw material in Sri Lankan apparel sector. This industry making use of cheaper imports from Pakistan, in turn exports apparel to destinations that include US and EU

Lessons Drawn From Pakistan-Sri Lanka FTA

There has been an increase in bilateral trade between Sri Lanka and Pakistan in the post – FTA milieu. Today Pakistan is the second largest trading partner of Sri Lanka amongst South Asian economies. There have been some concerns about the negative trade balance for Sri Lanka however the lower prices have resulted in an increase in welfare for both countries. The negative trade balance for Sri Lanka can also be justified on the account that a significant proportion of Sri Lankan imports from Pakistan constitute raw material and related intermediate inputs that in turn lead to efficiency gains. Most noticeable are the over 30 percent imports from Pakistan, currently being used as raw material in Sri Lankan apparel sector. This industry making use of cheaper imports from Pakistan, in turn exports apparel to destinations that include US and EU

Modulations of cell cycle checkpoints during HCV associated disease

Background Impaired proliferation of hepatocytes has been reported in chronic Hepatitis C virus infection. Considering the fundamental role played by cell cycle proteins in controlling cell proliferation, altered regulation of these proteins could significantly contribute to HCV disease progression and subsequent hepatocellular carcinoma (HCC). This study aimed to identify the alterations in cell cycle genes expression with respect to early and advanced disease of chronic HCV infection. Methods Using freshly frozen liver biopsies, mRNA levels of 84 cell cycle genes in pooled RNA samples from patients with early or advanced fibrosis of chronic HCV infection were studied. To associate mRNA levels with respective protein levels, four genes (p27, p15, KNTC1 and MAD2L1) with significant changes in mRNA levels (\u3e 2-fold, p-value \u3c 0.05) were selected, and their protein expressions were examined in the liver biopsies of 38 chronic hepatitis C patients. Results In the early fibrosis group, increased mRNA levels of cell proliferation genes as well as cell cycle inhibitor genes were observed. In the advanced fibrosis group, DNA damage response genes were up-regulated while those associated with chromosomal stability were down-regulated. Increased expression of CDK inhibitor protein p27 was consistent with its mRNA level detected in early group while the same was found to be negatively associated with liver fibrosis. CDK inhibitor protein p15 was highly expressed in both early and advanced group, but showed no correlation with fibrosis. Among the mitotic checkpoint regulators, expression of KNTC1 was significantly reduced in advanced group while MAD2L1 showed a non-significant decrease. Conclusion Collectively these results are suggestive of a disrupted cell cycle regulation in HCV-infected liver. The information presented here highlights the potential of identified proteins as predictive factors to identify patients with high risk of cell transformation and HCC development

Cell cycle modulations during Hepatitis C Virus associated disease progression

The disease burden caused by hepatitis C virus (HCV) infection entails a serious public health problem. Chronic infection causes liver inflammation and fibrosis, which often leads to liver cirrhosis and hepatocellular carcinoma (HCC). Increasing scientific evidence has suggested an impaired proliferation of hepatocytes during the course of HCV-associated pathogenesis. Considering the fundamental role played by cell cycle proteins in controlling cell proliferation and survival, altered regulation of these proteins could significantly contribute to HCV disease progression and subsequently, HCC. The aim of the work described in this thesis was to identify cell cycle modulations in early and advanced stages (pre-cancerous stages) of HCV-associated disease. In particular we wanted to explore the contribution of cell cycle genes in disease progression. Genome wide expression profiling was conducted on pooled RNA samples from HCV infected liver specimens with early (fibrosis stage 0-1) and advanced (fibrosis stage 3-4) disease stages. The results of this analysis reflect the changes taking place during the transition from early to advanced liver fibrosis, when the liver function becomes impaired and extracellular matrix deposition increases. In addition, it also showed altered expression of genes with functions in cancer development, cell growth and proliferation and cell death. Investigation of cell cycle alterations was the next target of this study. For this purpose, a comprehensive analysis of cell cycle pathway was performed on pooled RNA samples using Q RT-PCR. The results characterized early HCV (E-HCV) disease with increased expression of proliferation genes accompanied by elevated levels of cell cycle inhibitors possibly as a response to DNA damage or other cellular injuries. In advanced HCV (AHCV), DNA damage response genes and CDK inhibitors were found up-regulated. This was also accompanied by the down-regulation of genes involved in DNA repair and chromosomal stability. These results are suggestive of aberrant DNA replication and cell division that increases the risk of transformation and subsequent development of HCC. Post transcriptional regulation plays an important role in cell cycle proteins function. We further evaluated the protein expression of seven differentially expressed genes that were selected on the basis of their crucial role in cell cycle checkpoints. Consistent with the mRNA levels elevated expression of proliferation marker Mcm-2 and CDK inhibitors p27 and p27 were observed in hepatocytes of chronic hepatitis C patients. Moreover, expression of Mcm-2, p2l and p27 significantly associate with the progression of liver fibrosis. Expression of p53 was also elevated with no association to p2l expression. Among the mitotic checkpoint regulators, increased expression of KNTC1 and MAD2L1 proteins were found in E-HCV while in A-HCV KNTC1 expression significantly reduced and MAD2LI showed a non-significant decrease. This might suggest loss of their function in A-HCV. The analysis of apoptotic protein Caspase-3 revealed that its expression was mostly confined to sinusoidal lining cells and little in hepatocytes. Expression of anti-apoptotic protein Bcl-2 was negligible in hepatocytes and principally detected in infiltrating lymphocytes suggesting that the reduced apoptosis in hepatocytes might not be due to the anti-apoptotic activity of Bcl-2. Together these results are suggestive of an altered expression of cell cycle regulators in hepatocytes of HCV infected patients that could not only impair the regenerative potential of liver but could also make these cells susceptible to acquire mutations resulting in cell transformation and development of HCC

Diagnostic accuracy of various immunochromatographic tests for NS1 antigen and IgM antibodies detection in acute dengue virus infection

Introduction: Rapid diagnostic tests (RDTs) were evaluated, in this paper, for their utility as a reliable test, using resource-constrained studies. In most studies, NS1 antigen and immunoglobulin M (IgM)-based immunochromatographic tests (ICTs) were considered for acute phase detection. We aimed to evaluate the diagnostic accuracy of NS1, IgM, and NS1/IgM-based ICTs to detect acute dengue virus (DENV) infection in dengue-endemic regions.Methods: Studies were electronically identified using the following databases: MEDLINE, Embase, Cochrane Library, Web of Science, and CINAHL Plus. Keywords including dengue, rapid diagnostic test, immunochromatography, sensitivity, specificity, and diagnosis were applied across databases. In total, 15 studies were included. Quality assessment of the included studies was performed using the QUADAS-2 tool. All statistical analyses were conducted using RevMan, MedCalc, and SPSS software.Results: The studies revealed a total of 4135 individuals, originating largely from the Americas and Asia. The prevalence of DENV cases was 53.8%. Pooled sensitivities vs. specificities for NS1 (only), IgM (only) and combined NS1/IgM were 70.97% vs. 94.73%, 40.32% vs. 93.01%, and 78.62% vs. 88.47%, respectively. Diagnostic odds ratio (DOR) of DENV for NS1 ICTs was 43.95 (95% CI: 36.61-52.78), for IgM only ICTs was 8.99 (95% CI: 7.25-11.16), and for NS1/IgM ICTs was 28.22 (95% CI: 24.18-32.95). ELISA ICTs yielded a DOR of 21.36, 95% CI: 17.08-26.741. RT-PCR had a DOR of 40.43, 95% CI: 23.3-71.2. Heterogeneity tests for subgroup analysis by ICT manufacturers for NS1 ICTs revealed an χ2 finding of 158.818 (df = 8), p \u3c 0.001, whereas for IgM ICTs, the χ2 finding was 21.698 (df = 5), p \u3c 0.001.Conclusion: NS1-based ICTs had the highest diagnostic accuracy in acute phases of DENV infection. Certain factors influenced the pooled sensitivity, including ICT manufacturers, nature of the infection, reference method (RT-PCR), and serotypes. Prospective studies may examine the best strategy for incorporating ICTs for dengue diagnosis