ABSTRACT
Background: Discriminating latent tuberculosis infection [LTBI] from active TBI may be challenging. The objective of this study was to produce the recombinant L-alanine dehydrogenase [AlaDH] antigen and evaluate individuals with LTBI, those with active TBI, and uninfected individuals by enzyme-linked immunospot assay [ELISPOT] in order to distinguish LTBI from active TBI
Methods: This exploratory study was performed in the Iranian city of Shiraz from 2014 to 2015. The study population [N=99] was divided into 3 groups: individuals with newly diagnosed active TBI [n=33], their household contacts [n=33], and controls [n=33]. AlaDH was produced through PCR and cloning methods. The diagnostic characteristics of AlaDH vs. ESAT-6/ CFP-10 were evaluated in responses to interferon-gamma [IFN-gamma] and interleukin-2 [IL-2] with ELISPOT. Differences between the groups were assessed with the Kruskal-Wallis and Mann- Whitney tests for nonparametric data analysis. The statistical analyses were performed with SPSS, version 16
Results: IFN-gamma responses to both ESAT-6/CFP-10 [P=0.81] and AlaDH [P=0.18] revealed that there were no significant differences between the individuals with LTBI and those with active TBI. The same results were determined for IL-2 responses to ESAT-6/CFP-10 between the 2 groups, while significantly higher IL-2 responses to AlaDH were observed in LTBI than in active TBI. According to the ROC curve analysis, a cutoff value of 275 SFC showed sensitivity of 75.8% and specificity of 78.8% for distinguishing LTBI from active TBI by IL-2 responses to AlaDH
Conclusion: The current study suggests that it may be possible to discriminate LTBI from active TBI by IL-2 responses to AlaDH
ABSTRACT
Background: Dietary polyphenols, such as those found in green tea and red wine, are linked to antitumor activity. They are known to influence many signaling pathways epigenetically within the human body. In this regard, CPUK02 [15-Oxosteviol benzyl ester] is a new ent-kaurenoid derivative of stevioside and exhibits strong anti-cancer activity in vitro and in vivo. Nowadays, the role of epigenetics in cancer has been the subject of intensive study and DNA methylation targeting represents a relevant strategy for cancer treatment. There are no reports regarding the effects of CPUK02 on epigenetic alterations in colorectal cancer cell line. This study was an attempt to compare CPUK02 with 5-AZA as DNMT inhibitor agent and evaluate whether it can induce its anti-cancer effects via altering the level of DNMT3b mRNA, MGMT and SFRP2 methylation pattern in HCT 116 cell line
Methods: To evaluate DNMT3b expression, DNMT3B mRNA levels in HCT116 CRC cell line were quantified by real-time reverse-transcriptase Polymerase Chain Reaction [PCR] assay after 24 hr of incubation time with CPUK02 and 5-AZA. In addition, the methylation patterns of 2 CpG islands in this cell line were examined by methylation specific PCR methods
Results: CPUK02 surprisingly, decreased the DNMT3b mRNA level. The average expression levels of DNMT3b in HCT116 treated with CPUK02 and 5-AZA relative to the GAPDH expression level in control were 0.16 and 0.5%, respectively. Furthermore, CPUK02 could decrease the methylated allele of MGMT and SFRP2 genes in HCT 116 after 24 hr
Conclusion: In this study, positive correlation was found between mRNA expression of DNMT3b and gene promoter hypermethylation after treatment with CPUK02 and 5-AZA. Our data confirmed that CPUK02 like 5-AZA exhibits demethylating properties
Subject(s)
Humans , Epigenetic Repression/drug effects , Colorectal Neoplasms , Cell Line, Tumor/drug effects , Aza Compounds/adverse effectsABSTRACT
Epigenetics alterations, especially DNA methylation, play a critical role in control of gene expressions. Abnormal patterns of methylation are observed in early-stages of many cancers. Therefore, methylation analysis is useful in primary detection of tumors. Advances knowledge about the functional role of aberrant epigenetic modifications as potential biomarkers for cancer, have attracted considerable interest to pursue such investigations. Currently, many methodologies are available to distinguish methylation patterns, however, none is considered as the 'gold-standard' technique. This paper is an overview on some convenient methylation analysis methods
ABSTRACT
Background: Traditional medicines with anti-diabetic effects are considered suitable supplements to treat diabetes. Among medicinal herbs, Stevia Rebaudiana Bertoni is famous for its sweet taste and beneficial effect in regulation of glucose. However, little is known about the exact mechanism of stevia in pancreatic tissue. Therefore, this study investigated the possible effects of stevia on pancreas in managing hyperglycemia seen in streptozotocin-induced Sprague-Dawley rats
Methods: Sprague-Dawley rats were divided into four groups including normoglycemic, diabetic and two more diabetic groups in which, one was treated with aquatic extract of stevia [400 mg/kg] and the other with pioglitazone [10 mg/kg] for the period of 28 days. After completion of the experimental duration, rats were dissected; blood samples and pancreas were further used for detecting biochemical and histopathological changes. FBS, TG, cholestrol, HDL, LDL, ALT and AST levels were measured in sera. Moreover, MDA [malondialdehyde] level, catalase activity, levels of insulin and PPARgamma mRNA expression were also measured in pancreatic tissue
Results: Aquatic extract of stevia significantly reduced the FBS, triglycerides, MDA, ALT, AST levels and normalized catalase activity in treated rats compared with diabetic rats [p<0.05]. In addition to this, stevia surprisingly, increased PPARgamma and insulin mRNA levels in treated rats [p<0.05]. Furthermore, stevia compensated for the histopathological damage in diabetic rats
Conclusion: It is concluded that stevia acts on pancreatic tissue to elevate the insulin level and exerts beneficial anti-hyperglycemic effects through the PPARgamma-dependent mechanism and stevia's antioxidant properties
ABSTRACT
Background: Optimized RNA extraction from tissues and cell lines consists of four main stages regardless of the method of extraction: 1] homogenizing, 2] effective denaturation of proteins from RNA, 3] inactivation of ribonuclease, and 4] removal of any DNA, protein, and carbohydrate contamination. Isolation of undamaged intact RNA is challenging when the related tissue contains high levels of RNase. Various technical difficulties occur during extraction of RNA from pancreatic tissue due to spontaneous autolysis. Since standard routine protocols yield unacceptable results in pancrease, we have designed a simple method for RNA extraction by comparing different protocols
Methods: We obtained 20-30 mg pancreatic tissues in less than 2 min from 30 rats. Several methods were performed to extract RNA from pancreatic tissue and evaluate its integrity. All methods were performed three times to obtain reproducible results
Results: Immersing pancreatic tissue in RNA-later for 24 h at -80[degree sign]C yielded high quality RNA by using the TriPure reagent which was comparable to the commercial RNeasy Micro Kit. The quality of RNA was evaluated by spectrophotometer, electrophoresis and RT-PCR. We separated intact 28S and 18S ribosomal RNA [rRNA] when our procedure was compared with the RNeasy Micro Kit. Finally, full length of the actin gene was amplified by RT-PCR
Conclusion: We designed a simple, fast, cost-effective method for complete RNA extraction from the least amount of quantitatively intact pancreatic tissue