Expression of COLLAGEN 1 and ELASTIN genes in mitral valvular interstitial cells within microfiber reinforced hydrogel

Objective: The incidence of heart valve disease is increasing worldwide and the number of heart valve replacements is expected to increase in the future. By mimicking the main tissue structures and properties of heart valve, tissue engineering offers new options for the replacements. Applying an appropriate scaffold in fabricating tissue-engineered heart valves [TEHVs] is of importance since it affects the secretion of the main extracellular matrix [ECM] components, collagen 1 and elastin, which are crucial in providing the proper mechanical properties of TEHVs

Materials and Methods: Using real-time polymerase chain reaction [PCR] in this experimental study, the relative expression levels of COLLAGEN 1 and ELASTIN were obtained for three samples of each examined sheep mitral valvular interstitial cells [MVICs]-seeded onto electrospun poly [glycerol sebacate] [PGS]-poly [?-caprolactone] [PCL] microfi-brous, gelatin and hyaluronic acid based hydrogel-only and composite [PGS-PCL/hydrogel] scaffolds. This composite has been shown to create a synthetic three-dimensional [3D] microenvironment with appropriate mechanical and biological properties for MVICs

Results: Cell viability and metabolic activity were similar among all scaffold types. Our results showed that the level of relative expression of COLLAGEN 1 and ELASTIN genes was higher in the encapsulated composite scaffolds compared to PGS-PCL-only and hydrogel-only scaffolds with the difference being statistically significant [P<0.05]

Conclusion: The encapsulated composite scaffolds are more conducive to ECM secretion over the PGS-PCL-only and hydrogel-only scaffolds. This composite scaffold can serve as a model scaffold for heart valve tissue engineering

effect of MDMA and pentoxifylline drug on bad/bcl-xl gene dosage expression changes on rat liver

MDMA generally known as ecstasy, have deleterious effects on the serotonergic neurotransmitter system. Recent findings suggest that the liver and brain are major target organs of MDMA-related toxicities. Although most research is being dynamically performed on brain, however, the molecular mechanisms by which MDMA elicits adverse effects in both organs are poorly undrestood.The present study was performed to obtain evidence for molecular mechanism of apoptosis involved in MDMA-induced hepatotoxicity in rat liver after MDMAadministration. Moreover, the antagonistic effect of pentoxifylline was assessed on hepatotoxicity after MDMA administration. In this experimental study, sample size and power in each group were calculated as 10 rats with 95% confidence level and 5% confidence interval. In the study, four experimental groups were selected including Control Normal, MDMA, MDMA+PTX and PTX+MDMA. MDMA was dissolved in PBS and intraperitoneally injected three doses of 7.5mg/kg with two hours gap between doses. Pentoxyfilline also was injected as 100mg/kg, simultaneously with third dose of MDMA. After treatment, total RNA was isolated from liver tissue [5mg]. Absorbance at 260nm, 280nm and 230nm were measured and immediately reverse transcription was performed. Included target genes were BAD and BCL-XL as pro-apoptotic and anti-apoptotic gene, respectively. After set up and validation, Real-Time PCR were performed and obtaining data were statistically analyzed to determine significantly differences between groups. Using Real-Time quantitative PCR results, BCL-XL gene expression ratio significantly increased in MDMA+PTX group. Moreover, BAD gene expression ratio increased and up-regulated in PTX+MDMA group [P-value <0.001].Our study focused on molecular mechanism of MDMA in programmed cell death using gene expression quantification of a pro-apoptotic and anti-apoptoic gene in MDMA-induced hepatotoxocity. The results shown MDMA prompted apoptosis in liver and pentoxifylline protects hepatotoxicity after and befor taking MDMA

Microsatellite instability markers status in colorectal cancer

Colorectal cancer [CRC] is the third most prevalent and the third leading cause of cancer-related deaths in Iran. Our aim was to investigate five mononucleotide statuses among Iranian patients with sporadic colorectal cancer. In this experimental study investigation 80 sporadic CRC patients were evaluated for Microsatellite instability [MSI]. The pentaplex panel including 5 quasi mononucleotide microsatellite markers [NR-21, BAT-26, BAT-25, NR-27 and NR-24] was used. The MSI analysis was performed on paired tumoral DNA from cancerous tissues and genomic DNA from whole blood. MSI carriers were identified by analysis of tumor tissue using polymerase chain reaction. Our findings showed that microsatellite instability was detected in 36 of 80 cases [45%] with colorectal cancer. MSI analysis revealed that 17 cases of MSI-H [21%], 19 MSI-L [23%] and 44 MSS [55%]. Instability is observed in the tumoral DNA compared to the DNA from the normal DNA sample. The most instable markers were NR-21, NR-24 in which instability was detected in 45% of patients. Using a panel including 3 mentioned MSI markers should be more promising markers for identifying MSI status in patients with sporadic colorectal cancer

[Determination of gamma-globin induction levels by different concentrations of hydroxyurea in K 562 cell lines for beta thalassemia treatment

Hydroxyurea [HU] is currently used for beta thalassemia treatment through induction of fetal gamma-globin, In this study, effects of different concentrations of hydroxyurea on induction of gamma-globin gene in K562 cells, and inhibitory effect of siRNA against candidate gene which may be involved in HU mediated gamma-globin induction were assessed. In this eperimantal study, K562 cells were treated by different concentrations of HU [0, 50, 100 and 200 microM]. siRNA against candidate gene in HU mediated gamma-globin gene induction was transfected to K562 cells using lipofectamine 2000. The level of gammal-globin gene induction and inhibition were determined by quantitavie real-time PCR. There were 1.75-, 2.45- and 2.55- fold inductions in gamma-globin gene using 50, 100 and 200 microM HU, respectively. There has been 79.5% down regulation on candidate gene in siRNA transfected K562 cells. This study showed that gamma-globin induction in 100 microM HU is similar to 200 micro M HU treated K562 cells. There was also efficient inhibitory effect on candidate gene which may be involved in HU mediated gamma-globin induction

Sister chromatid exchange in peripheral blood lymphocytes as a possible breast cancer risk biomarker: a study of Iranian patients with breast cancer

Sister chromatid exchanges [SCEs] can be induced by various genotoxic treatments, suggesting that SCEs reflect a DNA repair process and it may be a good index for assessment of genomic instability. However, the occurrence of genetic instability and in particular, of spontaneous SCEs has been strongly linked to cancer. Several chromosomal regions and many genes have been implicated in breast cancer. Blood samples were obtained from 31 Iranian breast cancer patients and 11 healthy women. SCE was measured in peripheral blood lymphocytes by adding to Ham'sF10 medium in presence of PHA, BrdU [5-bromo-deoxy Uridine] fluorochrome Hoechst 33258, exposure to UV light and Giemsa staining. Then, SCE frequencies of patient and control groups were compared by the Mann-Withney U-test. Significantly difference was observed between two groups [p < 0.001]. This study indicates that SCE can be used as a risk biomarker for breast cancer