Ovarian stimulation by exogenous gonadotropin decreases the implantation rate and expression of mouse blastocysts integrins

Integrins are heterodimeric glycoprotein receptors that regulate the interaction of cells with extracellular matrix and may have a critical role in implantation. The aim of this study was to investigate the effect of ovulation induction on the expression of Alpha4, Alphav, Beta1, and Beta3 integrins in mouse blastocyst at the time of implantation. The ovarian stimulated and non-stimulated pregnant mice were sacrificed on the morning of 5[th] day of pregnancy. The blastocysts were collected, and the expression of Alphav, Alpha4, Beta1, and Beta3 integrins was examined using real-time RT-PCR and immunocytochemical techniques, then their ovarian hormones were analyzed at the same time. The implantation sites in uterine horns of other pregnant mice in both groups were determined under a stereomicroscope on the 7[th] day of pregnancy. The results showed that the expression of Alphav, Beta1, and Beta3 integrins in both mRNA and protein levels was significantly lower in the ovarian stimulated group than the control group, and the maximum ratio of expression was belonged to Beta1 molecule [P>0.05]. The implantation rate in superovulated mice was significantly lower than control mice. It was suggested that ovulation induction decreased the expression of Alphav, Beta1, and Beta3 integrins of mouse blastocysts

[Detection of HCMV infection in glioma brain tumors]

Human cytomegalovirus [HCMV] is a beta-herpesvirus that causes persistent infection in humans, as well as severe disease in fetuses and immunocompromised individuals. Although HCMV is not currently causally implicated in human cancer, emerging evidence suggests that HCMV infection may be specifically associated with malignancies such as gliomas. Gliomas are one of the most common brain tumors that affect humans. It is classified into four grades. In this study, we have developed and used a real-time PCR method for the detection and diagnosis of HCMV infection in glioma brain tumor samples. Paraffin-embedded tumor samples were chosen from patients who referred to Imam Khomeini Hospital Neurosurgery Ward. DNA was extracted from paraffinembedded tissues by a DNA extraction kit. After designing specific primers for the HCMV US28 region, a real-time PCR method was developed for detection of HCMV US28. The results of qualitative real-time PCR on 4/18 patients [22.2%] were positive. Two patients with positive HCMV results died. This is the first study that has monitored HCMV genes in samples from glioma patients in Iran. Considering the results of this study and controversies associated with other studies, a more comprehensive study using this and other diagnostic methods is suggested.

Comparison of qualitative PCR and pp65 antigenemia for the diagnosis of CMV infection in hematopoietic stem cell transplanted patients

Human cytomegalovirus [CMV] is a major life-threatening pathogen for hematopoietic stem cell transplant recipients. Specific tests are used for the diagnosis and monitoring of CMV infection in transplant patients. This study evaluates the performance of pp65 antigenemia and qualitative PCR assays for monitoring CMV in such patients. We analyzed 179 clinical samples from 41 patients by using a validated home-brewed qualitative PCR and a commercial antigenemia assay. The obtained results were evaluated using quantitative real-time PCR as the gold standard. CMV was observed in 26.8% of samples analyzed by the antigenemia assay and in 42.6% of the samples by qualitative PCR. Among 179 clinical samples, 50.8% were negative and 21.2% were positive by both assays. On the other hand, 26.3% were only positive by qualitative PCR whereas 1.7% were positive by the antigenemia assay. A comparison of the results with real-time PCR showed that qualitative PCR has a higher sensitivity than the antigenemia assay [98.7% vs. 45.7%]. The specificity of both assays was equal [96.8%]. Quantitative results of the antigenemia assay showed good correlation with real-time PCR [r=0.715; p<0.001] Both the qualitative PCR and antigenemia assays have special deficiencies for efficient diagnosis of CMV infection. Therefore, effective management of CMV infection in transplant patients requires the use of other sensitive quantitative methods such as qPCR

Localization of herpes simplex virus type 1 DNA in latently infected BALB/c mice neurons using in situ polymerase chain reaction

Herpes simplex virus type-1 [HSV-1] establishes a lifelong latent infection in neurons following primary infection. The existence of latent HSV-1 DNA in the trigeminal ganglia of infected BALB/c mice was examined using a direct in situ PCR technique, based on Digoxigenin-11-dUTP detection system with anti-digoxigenin-peroxidase and 3,3f-diaminobenzidine [DAB] substrate. Eight-week-old male BALB/c mice were inoculated via the eye by 10[4] plaque forming unit of wild type Iranian isolates of HSV-1. After establishment of latency, trigeminal ganglia were removed and examined using in situ PCR to detect HSV-1 genome. Finally, the results of in situ PCR were verified by a two-round PCR method, using amplification cocktail of in situ reaction, as a template for a conventional gel base PCR. The results suggest that a direct in situ PCR method using a peroxidase and DAB detection system is a useful means for detection of latent HSV-1 DNA in the latently infected ganglia