MASALAH-MASALAH PEMBELAJARAN YANG DIHADAPI WIDYAISWARA : Studi Kasus Pada Lembaga Diktat Pemda Tk.I Propinsi Bengkulu

<div><p>Rat strains differ dramatically in their susceptibility to mammary carcinogenesis. On the assumption that susceptibility genes are conserved across mammalian species and hence inform human carcinogenesis, numerous investigators have used genetic linkage studies in rats to identify genes responsible for differential susceptibility to carcinogenesis. Using a genetic backcross between the resistant Copenhagen (Cop) and susceptible Fischer 344 (F344) strains, we mapped a novel mammary carcinoma susceptibility (<i>Mcs30</i>) locus to the centromeric region on chromosome 12 (LOD score of ∼8.6 at the D12Rat59 marker). The <i>Mcs30</i> locus comprises approximately 12 Mbp on the long arm of rat RNO12 whose synteny is conserved on human chromosome 13q12 to 13q13. After analyzing numerous genes comprising this locus, we identified <i>Fry</i>, the rat ortholog of the furry gene of <i>Drosophila melanogaster,</i> as a candidate <i>Mcs</i> gene. We cloned and determined the complete nucleotide sequence of the 13 kbp <i>Fry</i> mRNA. Sequence analysis indicated that the <i>Fry</i> gene was highly conserved across evolution, with 90% similarity of the predicted amino acid sequence among eutherian mammals. Comparison of the <i>Fry</i> sequence in the Cop and F344 strains identified two non-synonymous single nucleotide polymorphisms (SNPs), one of which creates a putative, de novo phosphorylation site. Further analysis showed that the expression of the <i>Fry</i> gene is reduced in a majority of rat mammary tumors. Our results also suggested that FRY activity was reduced in human breast carcinoma cell lines as a result of reduced levels or mutation. This study is the first to identify the <i>Fry</i> gene as a candidate <i>Mcs</i> gene. Our data suggest that the SNPs within the <i>Fry</i> gene contribute to the genetic susceptibility of the F344 rat strain to mammary carcinogenesis. These results provide the foundation for analyzing the role of the human <i>FRY</i> gene in cancer susceptibility and progression.</p></div

A meta-analytical comparison of atenolol with angiotensin-converting enzyme inhibitors on arterial stiffness, peripheral blood pressure and heart rate in hypertensive patients

<p><i>Objectives</i>: This meta-analysis of randomized parallel controlled trials was designed to compare the efficacy of atenolol with angiotensin-converting enzyme inhibitors (ACEIs) in changing pulse wave velocity (PWV), peripheral blood pressure and heart rate (HR) among patients with essential hypertension. <i>Methods</i>: This study was conducted according to the PRISMA guideline. Data collection was independently completed by two investigators. Statistical analyses were completed by Stata software (v12.0). <i>Results</i>: Eight clinical trials were meta-analyzed in this study. Overall changes in PWV (weighted mean difference or WMD = 0.068, 95% confidence interval or CI: −0.487 to –0.623, P = 0.811) and peripheral systolic blood pressure (PSBP) (WMD = −1.281 mmHg, 95% CI: −6.936 to 4.375, P = 0.657) did not differ significantly between atenolol and ACEIs treatment. Relative to ACEIs, atenolol had a more favorable impact on peripheral diastolic blood pressure (PDBP) (WMD = −1.912 mmHg, 95% CI: −3.732 to −0.091, P = 0.040) and HR (WMD = −9.23 bpm, 95% CI: −12.53 to −5.93, P < 0.001). In stratified analyses, particularly by follow-up period, atenolol was observed to be superior over ACEIs within early 3-month treatment in PSBP (WMD = −4.097 mmHg, 95% CI: −6.589 to −1.605, P = 0.001), PDBP (WMD = −6.802 mmHg, 95% CI: −8.517 to −5.087, P < 0.001) and HR (WMD = −14.242 bpm, 95% CI: −16.427 to −12.058, P = 0.028), without heterogeneity (<i>I</i>² = 0.0%). There were low probabilities of publication bias for all comparisons. <i>Conclusions</i>: Our findings demonstrate that atenolol and ACEIs were equally effective in reducing PWV and PSBP, while atenolol was superior over ACEIs in improving PDBP and HR, especially within short-term treatment.</p

Lowered circulating apelin is significantly associated with an increased risk for hypertension: A meta-analysis

<p><i>Background and Objective</i>: Apelin is a bioactive peptide manifesting a potent vasodilatory property. In this meta-analysis, we aimed to investigate for the first time whether circulating apelin differed significantly between hypertensive patients and normotensive controls. <i>Methods</i>: Both PubMed and Embase were searched for eligible articles. Eligibility evaluation and data collection were done independently by two investigators. Weighted mean difference (WMD) with 95% confidence interval (CI) was calculated under random-effects model by STATA. <i>Results</i>: Ten studies were synthesized finally, including 1610 patients and 1105 controls. Overall analysis revealed that circulating apelin was significantly lowered in patients than in controls (WMD = −39.85 pg/mL, 95% CI: −65.56 to −14.15; P = 0.002), with significant heterogeneity (<i>I</i>² = 89.4%). By race, patients had lower circulating apelin than controls in Caucasian populations (WMD = −79.82 pg/mL, 95% CI: −105.80 to −53.85; P < 0.001), without heterogeneity (<i>I</i>²=0.0%), while no significance was observed in Chinese and African-Americans. Further grouping studies by source of controls found a significant reduction in circulating apelin in studies with hospital-based controls (WMD = −96.28 pg/mL, 95% CI: −129.67 to −62.88; P < 0.001) (<i>I</i>² = 49.4%), but not in studies with population-based controls. <i>Conclusions</i>: Via a meta-analysis of 10 studies and on 2715 subjects, our findings demonstrated that lowered circulating apelin was significantly associated with an increased risk for hypertension, especially in Caucasian populations.</p

<em>miR-205</em> Expression Promotes Cell Proliferation and Migration of Human Cervical Cancer Cells

<div><p>MicroRNAs (miRNAs) are short non-coding RNA regulators that control gene expression mainly through post-transcriptional silencing. We previously identified <em>miR-205</em> in a signature for human cervical cancer using a deep sequencing approach. In this study, we confirmed that <em>miR-205</em> expression was frequently higher in human cervical cancer than their matched normal tissue samples. Functionally, we demonstrate that <em>miR-205</em> promotes cell proliferation and migration in human cervical cancer cells. To further understand the biological roles of <em>miR-205</em>, we performed <em>in vivo</em> crosslinking and Argonaute 2 immunoprecipitation of miRNA ribonucleoprotein complexes followed by microarray analysis (CLIP-Chip) to identify its potential mRNA targets. Applying CLIP-Chip on gain- and loss-of-function experiments, we identified a set of transcripts as potential targets of <em>miR-205</em>. Several targets are functionally involved in cellular proliferation and migration. Two of them, CYR61 and CTGF, were further validated by Western blot analysis and quantification of mRNA enrichment in the Ago2 immunoprecipitates using qRT-PCR. Furthermore, both <em>CYR61</em> and <em>CTGF</em> were downregulated in cervical cancer tissues. In summary, our findings reveal novel functional roles and targets of <em>miR-205</em> in human cervical cancer, which may provide new insights about its role in cervical carcinogenesis and its potential value for clinical diagnosis.</p> </div

Real time quantitative RT-PCR of <i>miR-205</i> expression in human cervical tumors, normal cervices and cervical cancer cell lines, normalized to the geometric mean of <i>RNU6B</i> and <i>RNU43</i>.

<p>(A) <i>miR-205</i> expression was significantly higher in the tumors than the normal samples (<i>P</i><0.001; paired t-test). (B) Relatively higher expression of <i>miR-205</i> was found in a majority of tumor samples as compared to their normal counterparts. (C) High expression of <i>miR-205</i> was detected in ME-180, C4I and CaSki cells, and low or undetectable expression level was found in HeLa, SW756, SiHa and C33A cells. Data presented represent mean of three independent experiments with triplicates. Error bars represent standard deviations from the mean.</p

<i>CYR61</i> and <i>CTGF</i> mRNAs expression in human cervical samples, and their correlations with <i>miR-205</i> expression.

<p>Relatively lower expression of <i>CYR61</i> (A) and <i>CTGF</i> (B) was found in a majority of tumor samples as compared to their normal counterparts (n = 28). (C) The expression of <i>CYR61</i> and <i>CTGF</i> was significantly lower in the tumors than the normal samples (<i>P</i> = 0.002 and <i>P</i><0.001, respectively; paired t-test). (D) Inverse correlation between the expression level of <i>miR-205</i> and <i>CYR61</i> (upper) or <i>CTGF</i> (lower). The expression relationship was evaluated by Pearson’s correlation analysis. <i>P</i><0.05 was considered statistically significant.</p

Evaluation of <i>CYR61</i> and <i>CTGF</i> as targets of <i>miR-205</i>.

<p>(A) Representative Western blot showing the protein expression levels of CYR61 and CTGF in cells transfected with a <i>miR-205</i> mimic, <i>miR-205</i> inhibitor, or corresponding scramble and mock transfection controls. (B) CYR61 protein expression was significantly repressed in <i>miR-205</i>-overexpressing (treated with Pre-miR-205) cells and significantly increased in <i>miR-205</i>-depleting (treated with Anti-miR-205) cells as compared to their respective negative controls. CTGF protein expression was slightly repressed in HeLa cells treated with Pre-miR-205, and slightly increased in CaSki cells treated with Anti-miR-205, but the effect was not statistically significant. Data presented represent mean of at least four independent experiments. qRT-PCR analysis of <i>CYR61</i> (C) and <i>CTGF</i> (D) mRNA in the Ago2-immunoprecipitated RNAs of <i>miR-205</i>-overexpressing or -depleted cells as compared to mock-transfection control. Relative expression level of individual mRNAs was normalized to <i>miR-21</i> expression (as endogenous control for Ago2 IP RNA). Fold change was calculated by dividing the normalized expression values of Ago2-immunoprecipiated samples by the normalized expression values of its respective input samples. Data presented represent mean of at least three independent experiments. Error bars represent standard deviations from the mean. All comparisons were evaluated using <i>t</i>-test. *<i>P</i><0.05; **<i>P</i><0.01; ***<i>P</i><0.001; <i>n.s.</i> = not significant.</p

Functional analyses of <i>miR-205</i> regulation in cervical cancer cell lines.

<p>(A) Cell proliferation was assessed in human cervical cancer cell lines transfected with a <i>miR-205</i> mimic (Pre-miR-205), inhibitor (Anti-miR-205) or corresponding negative control (Anti-miR Neg control or Pre-miR Neg control) using WST-1 assay. Relative cell growth was normalized to its respective control-treated cells. (B) Graphs showing relative cell migration in both <i>miR-205</i> inhibition and overexpression experiments as evaluated by Transwell migration assay. (C) Representative images of cell migration evaluated by wound healing assay. Scratch wounds were made on confluent monolayer cultures after 48 h of transfection. Images of wound repair were taken at 0, 18 and 24 h after wound (left panel). The percentage of wound closure was normalized by wound area at 0 h (right panel). Data presented represent mean of three independent experiments. Error bars represent standard deviations from the mean. All comparisons were evaluated using t-test. *<i>P</i><0.05; **<i>P</i><0.01; ***<i>P</i><0.001; <i>n.s.</i> = not significant.</p

Comparison of the catestatin levels between adverse events group and non-adverse events group.

<p>Box plots showing the distribution of concentrations of plasma catestatin in two groups (n1 = 26, n2 = 74) on ER (A), D3 (B) and D7 (C). Boxes specify inter-quartile ranges; the horizontal bars inside the boxes indicate medians; the upper and lower whiskers get to the most distant value within the 95% of the distribution. The p-values for the comparison of the groups are also indicated.</p

Patient characteristics according to occurrence of adverse events.

<p>Values represent mean ± SD or the percent of the patients.</p><p>LVEDD, left ventricular end diastolic diameter; LVEF, left ventricular ejection fraction; LAD, left anterior descending coronary artery; LCX, left circumflex coronary artery; RCA, right coronary artery</p><p>Patient characteristics according to occurrence of adverse events.</p