Downregulation of Sirt1 is correlated to upregulation of p53 and increased apoptosis in epicardial adipose tissue of patients with coronary artery disease

The higher expression level of p53 in epithelial adipose tissue (EAT) has previously been reported in atherosclerosis. Since we hypothesized that the expression of p53 is modulated by Sirt1, the aim of this study was to determine the expression levels of Sirt1 and p53 and to investigate their correlation to apoptosis in EAT of patients with coronary artery disease (CAD). Thirty-five patients with more than 50 % stenosis in at least one of the main coronary arteries were considered as CAD group while 29 patients with no clinical signs of atherosclerosis who underwent open-heart surgery for valve replacement were classified as control group. EAT biopsy samples were collected from all participants during surgery. Sirt1, p53, Bax, and Bcl‑2 gene expression levels were determined in EAT by qRT-PCR and Western blotting was carried out to assess Sirt1 and p53 protein levels. Hematoxylin and eosin staining was used for histopathological analysis. mRNA and protein levels of Sirt1 in EAT were significantly lower in patients with CAD compared with control group, whereas CAD patients showed greater p53 gene and protein expressions. In addition, inverse correlations were observed between Sirt1 and p53 at both mRNA and protein levels. The Bax and ratio of Bax/Bcl-2 gene expressions were higher in CAD group, but no difference was observed in Bcl-2 expression. Histopathological analysis showed apoptotic bodies and infiltrated immune cells in EAT of CAD group. Our results suggest that the Sirt1-p53 axis may involve in atherosclerosis by promotion of apoptosis

Expression of Integrin β1, Focal Adhesion Kinase, and PDZ-Binding Motif in Human Liver Cirrhosis and Simple Steatosis

Background: Integrins are transmembrane mechanosensitive proteins that negatively contribute to the pathogenesis of different types of chronic liver disease and can activate focal adhesion kinase (FAK). Objectives: This study aimed to determine the hepatic integrin β1 and FAK mRNA as well as the transcriptional coactivator with PDZ-binding motif (TAZ) protein expressions in cirrhotic patients and simple steatosis. Methods: In this case–control study, liver tissues were collected from 30 cirrhotic patients with various etiologies (i.e., nonalcoholic steatohepatitis-, primary sclerosing cholangitis-, alcoholic-, autoimmune hepatitis [AIH]- and hepatitis B virus [HBV]/hepatitis C virus [HCV]-related cirrhosis [six per group]), liver samples with simple steatosis (n=6), and control liver tissues (n=9). Results: Integrin β1 gene expression was significantly up-regulated in all cirrhotic groups compared to control group (P<0.05), with the exception of AIH cirrhosis. However, hepatic FAK gene expression and TAZ protein level in the cirrhotic groups were not significantly different than those in the control group. Furthermore, hepatic integrin β1 and FAK gene expressions as well as TAZ protein level in simple steatosis were significantly lower than those in nonalcoholic steatohepatitis (NASH) cirrhosis and control (P<0.05). Conclusion: Integrin β1 was up-regulated in cirrhotic liver tissues. In addition, FAK, integrin β1, and TAZ were concordantly down-regulated in simple steatosis, and may have been involve in the steatosis development

Attenuation of Differentiation of Mesenchymal Stem Cells Into Adipocytes by Co-administration of Calcium and All-trans Retinoic Acid

Background: Adipogenesis is affected by multiple factors, among which all-trans retinoic acid (ATRA) and Ca2+are considered important factors. Objectives: This study aimed to evaluate the effect of calcium, ATRA, and their underlying molecular mechanisms, alone and in combination, on adipocyte differentiation. Methods: Human adipose-derived mesenchymal stem cells (hAD-MSCs) were differentiated into adipocytes and simultaneously exposed to 0.5 μM ATRA or 2.5 mM calcium, or both in combination for 14 days. Results: Higher intracellular Ca2+was observed in both Ca2+and Ca2+plus ATRA groups. Assessment of triglyceride content and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) activity indicated lower differentiation levels in all treatment groups than in the control group. Furthermore, the results demonstrated a higher expression of retinoic acid receptor alpha (RAR-α) and lower expression of peroxisome proliferator-activated receptor γ2 (PPARγ2) and glucose transferase-4 (GLUT4) in the treatment groups as compared with the control group. It is noteworthy that Ca2+plus ATRA treatment caused more significant effects on gene expression levels (P<0.05). Conclusion: In conclusion, combined treatment with Ca2+and ATRA has a more pronounced inhibitory effect on adipocyte differentiation, indicating their cumulative effect

Determination of Osteoblast Cell Viability and Histological Changes of Samples Obtained from Different Implant Drills during Osteotomy

Statement of the Problem: The bone particles collected during osteotomy could be used as autogenous bone graft materials for dental implant surgery. Different factors such as drill design may influence its clinical viability.Purpose: This study examined the effect of drill design on the osteoblast viability and histopathology parameters of bone collected during the preparation of dental implant site.Materials and Method: In this experimental study, 90 samples were obtained from three different bone drilling systems including Bego, Implantium, and Dio during fixture installation in patients requiring treatment at the Department of Periodontology, Dentistry University Hamedan. The MTT (3-4,5-Dimethylthiazol2,5-diphenyltetrazolium bromide) was used to determine percentage of cell viability. Samples were fixed in 10% formaldehyde for histological evaluation. Then, they were kept in 10% EDTA solution for 4 weeks for decalcification. The provided slides were evaluated regarding bone structure and osteocytes counts for assessment of viability. Tukey test and SPPS 21 software were used for statistical analysis.Results: The result showed the viability of osteoblast obtained by Dio (0.45±0.04) was significantly better than Bego (0.37±0.05) and Implantium (0.37±0.04) systems. In histopathological evaluation, the grafting material obtained by Dio presented the best osteoblast morphology.Conclusion: It might be concluded that drill geometry has significantly influenced the viability of bone particles collected during the preparation of implant sites .Moreover, characteristic geometry alone cannot represent the performance of a particular drill, and several geometric features should be concerned. The results of this study showed that the geometry of the Dio drill was the best considering the viability and histopathological evaluations

Characterisation of the P19CL6 cardiovascular cell-line at the gene expression level : effects of long-chain fatty acids

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Characterisation of the P19CL6 Cardiovascular Cell-Line at the Gene Expression Level: Effects of Long-Chain Fatty Acids.

Dietary fats have been implicated in the progression of coronary heart disease and clear differences have been shown for the effects of saturated, monounsaturated and polyunsaturated fatty acids on the different stages of disease. In part, these effects can be explained by the regulation of gene transcription. In this study, a novel cardiac cell-line was targeted for the investigation of fatty acid effects on gene transcription. The aims of this study were to (i) characterise the P19CL6 cell-line at the transcriptional level, (ii) investigate the effects of long-chain fatty acids and clofibrate on mRNA levels of specific lipid metabolism-related genes, such as heart-type fatty acid-binding protein and peroxisome proliferator-activated receptor-&alpha;, -&beta;, and -&gamma; in the P19CL6 cell-line, and (iii) determine the effects of long chain fatty acids and clofibrate on global transcriptome levels in P19CL6 cells, using cDNA microarray analysis. P19CL6 cells displayed characteristics indicative of a cardiomyocyte phenotype including: the expression of cardiac-specific markers (&alpha;-MHC, &beta;-MHC and CaCh-h), an increase in pulse rate with increasing adrenaline concentration and the display of mononuclear cardiomyocyte morphology. However, spontaneous contraction was inconsistent between cultures of P19CL6 cells and pulse rate decreased significantly with increasing passage number until no “beating” was eventually recorded. These observations led us to conclude that P19CL6 cells may display a heterogeneous phenotype, which was investigated further. Analysis of microarray data indicated that global transcriptome profiles observed in the P19CL6 cell-line are not indicative of a pure adult cardiac or skeletal muscle phenotype, and more closely related to mouse embryonic heart tissue. However, the H9C2(2-1) cell-line was more closely related to adult and embryonic heart tissue than the P19CL6 cell-line. Therefore it was concluded that the H9C2(2-1) cell-line represents a better 'cardiomyocyte model system than the P19CL6 cell-line. The results of this study also showed that in P19CL6 cells long-chain fatty acids (but not clofibrate) significantly increased the abundance of peroxisome proliferator-activated receptor-&alpha; and -&gamma;, whereas no changes were observed in the expression levels of heart-type fatty acid-binding protein and peroxisome proliferator-activated receptor-&beta;, under the conditions used. The non-response of the latter genes can probably be explained by the relatively short time of exposure of cells in culture. Microarray analysis showed that linoleic and &alpha;-linolenic acids and clofibrate had similar effects, and that these differed from those of palmitic and oleic acids. These results are in agreement with other studies showing that cellular responses to polyunsaturated fatty acids differ from those observed with saturated and monounsaturated fatty acids

The Evaluation of the effects of cytotoxic, anti-proliferative and anti-migrative effects of the thymoquinone on PC3 prostate cancer cell line

Background & Objective: Herbal medicine and their composition are widely used for different therapeutic purposes. Therefore, this study was designed to evaluate of the anti-proliferation and anti-migration effects of Thymoquinone (TQ), an active part of Nigella sativa, on PC3 prostate cancer cells. Materials & Methods: In this experimental study, PC3 prostate cancer cells were treated with 0, 10, 30, 50, 70 and 90 µM concentrations of thymoquinone for 12, 24 and 48 hours and cell viability was determined by MTT assay. IC50 concentration of thymoquinone was obtained as 40 µM concentration of thymoquinone and 24h incubation time. PC3 cell migration was studied using the cell migration assay. Colony formation assay was used to determine cell proliferation and Gimsa staining was performed to observe morphological changes of the cells. Results: The IC50 of Thymoquinone was found in concentration of 40 µM at 24h treatment. Thymoquinone significantly (p<0.05) inhibited cell growth and proliferation as well as cell migration. Treatment of PC3 cells with 40 µM thymoquinone showed significant changes in nucleus and cytoplasm of the cells which are representative of apoptosis and cell death. Conclusion: Thymoquinone has significant anti-proliferative and anti-migrative effects on prostate PC3 cells and therefore may be used as a complementary agent for prevention of prostate cancer

Influences of combining nano zinc, honey and Aloe vera to accelerate healing the wounds caused by third–degree burn in male balb/c mice

Introduction: Burns are one of the most common household and industrial injuries. There are evidences which demonstrate the therapeutic properties of honey and Aloe vera. We evaluated the topical influences of this material and nano zinc combination on healing the wounds caused by third-degree burns. Materials and methods: 32 balb/c mice divided into a control group (without treatment), group 1 (treated with Aloe vera and nano zinc), group 2 (treated with Aloe vera, honey and nanoz inc) and group 3 (treated with honey and nano zinc). The third-degree burn was created on the back of balb/c mice with general anesthesia observing sterile conditions. Local treatment of burn was conducted once a week during 6 weeks and after the end of treatment, were anesthetized by ether and then killed. After fixation, the practical steps of general histology technique were performed on it. The samples stained with hematoxylin–eosin and they observed with a microscope. Results: We found full tightening of the burn wound and less scar in the group treated with nano zinc and honey compared to control group and other groups. In histological studies, a significant increase was found in the overall thickness of the skin, keratinocyte layer, the epidermis and hypodermis, number and diameter of the hair follicles in a third group versus other groups. Conclusion: The results showed the organic honey and nano zinc combination accelerate the healing process of burn wound in male balb/c mice. While adding Aloe vera to this composition doesn't have an effect on wound healing