Research in Molecular Medicine
Volume:2 Issue: 4, Nov 2014

Myeloproliferative Neoplasm (MPN) are a clonal disorder in hematopoietic stem cells (HSC). MPN is categorized to 8 subclasses, including chronic myeloid leukemia (CML), polycythemia vera (PV), essential thrombocytopenia (ET), primary myelofibrosis (PMF), systematic mastositosis (SM), chronic eosinophilic leukemia (CEL), chronic neutrophilic leukemia (CNL), and unclassified myelofibrosis disorders (UMPN). It usually occurs in 5th to 7th decade of life. However, CNL and ET have been observed in children. A lot of mutations have been identified in these disorders that Jak2V617F is the most important mutation. Moreover, several somatic mutations other than JAK2V617F in MPN patients have been reported. Such mutations include MPL, TET2, ASXL1, IDH1, IDH2, CBL, LNK, IKZF, and EZH2 from precursor stem cells. The role of mutations mentioned is not clear in pathogenesis of this disease. Hence, in this study, mutations in different stages of myeloproliferative disorders have been reviewed.

Amplification of HER2 is seen in 20-30% of breast cancer cases. Measurement of HER2 gene amplification appears to be of vital importance in planning the treatment schedule for patients with breast carcinoma. The aim of our study was to evaluate HER2 amplification status in malignant and benign breast tumors by differential PCR (dPCR). The genomic DNA was extracted using the phenol/chloroform extraction procedure from 76 different breast tissues. Differential PCR was performed using the DNA samples isolated from fresh and paraffin- embedded breast cancer tissues. The relative copy number ratio of target gene (HER2) to control gene (INF-γ) was measured. dPCR products were then separated by electrophoresis using 2% agarose gel. The intensity of HER2 and INFγ bands were determined for each sample by ImageJ software. According to the ratio between the band intensity of HER2 to INFγ in tumour and also normal samples, 7% and 26% rates of HER2 amplification were observed in benign and malignant samples respectively. The ratio showed a 2-5 fold increase in HER2 gene copy number for tissues with HER2 amplification; whereas, a one-fold increase was found in other samples. Differential PCR provides a relatively rapid and inexpensive technique to assess the HER2 gene amplification, especially alongside immunohistochemistry as a routine assessing method.

Periodontal ligament stem cells (PDLSCs) are considered as a type of mesenchymal stem cell that is beneficial target for numerous clinical applications in periodontal tissue regeneration therapy. This study examined the effects of dexamethasone (Dex) on human PDLSCs in vitro. PDLSCs obtained from the roots of patient’s teeth were cultured with Dex (0.01 μM), and their proliferation was measured. The osteogenic differentiation was assessed by alkaline phosphatase (ALP) activity and Alizarin Red-S staining for calcium deposition. After the administration of 0.01 μM Dex, the activity of ALP increased significantly. Furthermore, mineralized nodule formation showing the intracellular calcium deposition was significantly higher in the Dex-treated cells than that of the control cells. Collectively, Dex has positive effects on osteogenic differentiation of human PDLSCs in vitro. It is suggested that PDLSCs may serve as a potential material for periodontal tissue regeneration.

Breast cancer is the most common malignancy in females worldwide. Several etiological factors including environmental factors have been recognized for breast cancer. Epstein Barr virus as a viral etiological factor has been proposed. So far, several studies have investigated the relationship between development of breast cancer and Epstein Barr virus, but few have been done in Iran. The aim of this study was to determine whether there was an association between EBV infection and female breast cancer in Iran. We analyzed paraffin embedded breast tissue specimens by polymerase chain reaction (PCR) including breast cancer specimens (as case group) and breast fibroadenoma specimens (as control group). PCR was performed to amplify specific sequences of EBV. From 130 cases of breast samples, 67 cases of breast cancer tissues and 41 cases of breast fibroadenoma tissues had adequate quality and quantity of DNA to detect EBV. PCR for EBV was positive in 4 invasive ductal carcinoma specimens (7.3%) and only one of the fibroadenoma specimens (2.4%). No significant association was found between EBV infection and invasive ductal carcinoma (p> 0.05). Also, patient’s age and histological grade of IDC were not correlated with EBV infection (p>0.05). We observed no etiologic association between EBV infection and invasive ductal carcinoma of female breast in our regions; however, further studies are required to elucidate this association.

Systemic-onset Juvenile Idiopathic Arthritis (SoJIA) is an autoinflammatory disease with complex genetic trait starts in children less than 16 years of age with fever and cutaneous rash. Despite, the main genetic factors that may play a role in SoJIA have not yet been identified. High level of interleukin-1beta in the blood of SoJIA patients has been reported. The production and secretion of IL-1 β is related to pyrin coded by mediterranean fever gene (MEFV gene). Therefore, mutation in MEFV may be associated with SoJIA diseases. This study aimed to identify the association between R202Q mutation in exon 2 of MEFV gene and SoJIA disease. This study was done in 30 SoJIA patients and 30 controls. DNA was extracted from blood cells and analyzed by RFLP-PCR. The PCR product was digested with PvuII and then separated by gel electrophoresis. R202Q mutation was found in 3.3% of control and 43.3% of patient group. Significant statistical differences were observed between cases and controls in the R202Q mutation. The present study showed that the mutation in MEFV gene is a susceptible factor in development of SoJIA disease in Iranian patients.

Cholera is a potentially epidemic and life-threatening secretory diarrhoea characterized by voluminous watery stools, often accompanied by vomiting, and resulting in hypovolemic shock and acidosis. It is caused by certain strains of the species Vibrio cholerae which can also cause mild or in apparent infections. The aim of this study is the evaluation of Capsaicin, as a potential inhibitor of zonula occludens toxin production in V. cholerae ATCC 14035. MIC of capsaicin was determined by Broth Microdilution method according to CLSI guidelines. The zot gene expression level were analysed using real-time RT–PCR technique. Results from MIC test showed that 100 μg mL−1of capsaicin was the highest concentration that did not affect the bacterial growth; however, zonula occludens toxin (zot) gene expression of the tested strain was significantly inhibited by capsaicin in a dose-dependent manner at sub-bacteriocidal concentrations. The recA gene did not show any significant difference in its expression with or without capsaicin. Capsaicin is one of the active compounds of red chili that can drastically suppress zot gene expression and shows promising inhibitory effect against V. cholerae zot production. Thus, routine intake of red chilli, which is easily available and inexpensive, may be an alternative approach to prevent and control symptoms of cholera.