Journal of Pathobiology Reaearch
Volume:14 Issue: 3, 2011

Estrogen receptor alpha protein status is determined by routine immunohistochemistry analysis in all malignant breast tumors. This assay has its limitations. RNA based techniques are potential complements for immunohistochemistry but it must be noticed that gene silencing may occur at different levels from RNA to protein. The aim of this study was the comparison of the results from these two assays and characterizing the tumors subgroup in which gene expression occurs at RNA level but the target protein is absent. 92 primary breast tumors including their clinical and IHC results were collected before treatment. Estrogen receptor gene expression of tumors was studied by Reverse Transcription Polymerase Chain Reaction (RT PCR). In this assay, GAPDH was used as a reference gene. 36.6 % of tumors with negative estrogen receptor protein showed gene expression at mRNA level. In this subgroup most of the patient were older than 50 years and in stages 3 or 4 of breast cancer and had poor prognosis according to Nottingham prognostic index. Most cases of the perineural invasion have been seen in this subgroup. It seems that RT-PCR assay would enable us to recognize a subgroup of breast tumors with poor prognosis which expresses RNA but not protein.

Microsporidia are ubiquitous opportunistic pathogens infecting all animal phyla. The purpose of this study was to characterization human-associated microsporidia in pigeons of Tehran by staining and molecular methods. In the year 2010 a total of 147 pigeon’s fecal samples were randomly collected from bird stores and public parks of Tehran and screened for the existence of human pathogenic microsporidia by staining method and multiplex/Nested-PCR and RFLP techniques. Nineteen (12.92%) of the studied samples were positive by microscopic examination, and 31 (21.08%) isolates were detected with specific primers. Genotyping based on the ITS regions of the rRNA gene were done for the Entrocytozoon bieneusi, Encephalitozoon intestinalis, Enc. hellem and Enc. cuniculi, respectively. The genotypes of Ent. bieneusi were identical to the D, M and J; genotypes of Enc. hellem were similar to the genotype 1 and 3 and genotypes of Enc. cuniculi were equal to I and II genotypes which previously characterized in human and animal origins. These results revealed that there is no limits to microsporidia transmission between pigeons of Tehran and humans for human infective species. This study points to the hygienic importance of this bird, because feces of pigeons are one of the sources of infection with microsporidia in human and easily pollutes our environment; on the other hand, children and elderly people comprise the principal visitors of public parks and they are the populations at risk for microsporidiosis.

Leishmaniasis is one of the six most common parasitic infections in tropical regions. There are different therapeutic modalities. However therapeutic resistance is developed and resulted in numerous problems. So evaluation of other therapeutic modalities is performed extensively. We compared the therapeutic response of cutaneous leishmaniasis with Glucantime and Garlic extract and itR10 in animal model. This experimental study was conducted in Shahed University. The therapeutic response of cutaneous leishmaniasis to Glucantime and Garlic extract and R10 in animal model was studied in BALB/c, outbred SW mice and C57BL/6 mice. These three races were divided in four groups according to receiving either one of these three agents or no treatment (control). The therapeutic response was evaluated according to parasitic load before and after treatment and also with measuring the size of the lesions. The results showed that R10 had good therapeutic efficacy in treatment of lesions in mice (P<0.05) that this efficacy was significant in sixth, seventh and eighth weeks after the treatment. There was also a statistically significant difference between the groups regarding the parasitic load (P<0.05). According to the results, it may be concluded that R10 extract would have a good efficacy in treatment of cutaneous leishmaniasis that is comparable with glucantime.

The aim of this study was to evaluate and compare the population of Natural killer T lymphocyte (NKT) in the uterus and spleen of the hyperstimulated and control mice at the seventh day of pregnancy. The superovulated and control mice were put individually in a cage with a male mouse. In the next morning they were considered for observing the vaginal plag. The day was assumed as the first day of pregnancy. On the 7th day of pregnancy, the samples were collected from uterus of implantation site, interval site and spleen tissues and 5 micrometer cryosections were prepared. The immunohistochemical reaction was used for CD 161 and CD3 markers and the distribution of NKT cells population were compared with nucleated cells in hyperstimulated and control groups. There were not significant differences between the NKT cell population of the spleen, decidual and myometrial tissue of interval site in the control and hyperstimulated groups. But this population was increased in the hyperstimulated group (2.26 ± 1.43) compared with control (0.79 ± 0.17; P≤0.05) in the decidual of implantation site, moreover there was not any significant difference at myometrial tissue of both groups in implantation site. It seems that ovulation induction could not affect systematically on the population of NKT cells of pregnant mice while it could locally cause an increase in the decidual NKT cells.

Mitochondria is a critical organelle within the cell and has important role in the oocyte development, thus the aim of this study was the comparison of mitochondrial distribution within the mouse oocyte at germinal vesicle (GV), germinal vesicle breakdown (GVBD) and metaphase II (MII) stages using laser confocal microscopy (LSCM). After superovulation of adult mouse (FVB/N) using 7.5 IU PMSG the oocytes at GV, GVBD and MII were collected mechanically from the ovary then they were stained with (1 microgram per 1 ml) 5,5´6,6´-tetrachloro-1,1,3,3´tetraethylbenz-imidazolycarbocyanine iodide or JC-1 for ten min at 37ºC. Then they were observed under LSCM at 515-530 nm wavelengths for green light. Our observation showed the mitochondria were distributed homogenously within the cytoplasm of GVBD and MII oocytes as green staining but they were heterogeneously within the GV oocytes and some of them were located around the oocyte nucleus as a ring. Our results showed two mitochondrial distribution patterns within the mouse oocytes at different developmental stages. It seems that the main factors for distribution of mitochondria within the cells is their need for producing ATP and usage of the energy.

Survey of the influence of HLA-DRB1, -DQB1 alleles, genotypes and haplotypes on age at onset of type 1 diabetes (T1D) in an Iranian population 105 Iranian T1D patients of different ethnic group and 100 ethnically, age and sex matched individuals were selected from Tehran's hospitals and HLA-DRB, -DQB typing was performed. According to the age at onset of T1D, the patients were divided into 4 groups (1-5, 6-10, 11-15, 16-20 years). The frequency of susceptible and protective alleles, genotypes and haplotypes was calculated in each group. The data were evaluated by using fisher's exact test. Odds Ratio or relative Risk was measured for all samples. The results illustrated that the frequency of the HLA-DRB1*0401 allele decreased with increasing age, whereas the frequency of the HLA-DQB1*0201 allele increased with increasing age. The HLA-DRB1*0301 and HLA-DQB1*0302 alleles demonstrated the highest frequency in the 6-11 and 1-5 years age at onset group, respectively. HLA-DRB1*0401-DQB1*0302 haplotype had the most frequency among the 1-5 years age at onset group (p: 2×10-7, OR: 69.919) and the frequency of HLA-DRB1*0301-DQB1*0201 haplotype was the highest in the 6-11 years age at onset group among others (p: 2×10-6, OR: 6.243). The current study indicated that HLA-DRB1, -DQB1 alleles, genotypes and haplotypes are associated with age at onset of type1 Diabetes in Iranian T1D patients. The individuals carrying alleles that are associated with younger age at onset should take care under preventive treatment.

Low density Lipo-protein Receptor- related Protein (LRP) is the most important cholesterol receptor in neurons. It serves as a receptor for APOE protein which is the most important risk factor for Alzheimer’s Disease. LRP also contributes to the ligation of lipoproteins with APOE in neurons. Association between LRP C766T and Alzheimer’s disease in Iranian patients with late onset Alzheimer’s disease (LOAD) was investigated in this research. 100 patients with LOAD were selected based on DSM-IV-TR and NINCDS-ADRDA diagnostic criteria and 100 normal controls without any personal and family history of Alzheimer’s disease or dementia were included in this case- control study. AD patients and control subjects were matched for age and sex. PCR-RFLP was set up to detect LRP C766T polymorphism. LRP C/C genotype and C allele distribution were more frequent in AD patients than in control subjects. However, this difference was not statistically significant. When association between LRP C/C genotype and AD was categorized by the gender, in both genders, there was not any significant correlation. Our findings indicate that 766C allele of LRP gene could not significantly alter the risk of developing late-onset Alzheimer's disease in Iranian patients. Analysis of other genetic factors and environmental factors are promoted in Iranian population.

The aim of this study was to construct a pcDNA3.1+ vector containing FMDV type O/IRN/1/2007-VP1 gene, protein expression in BHKT7 cells and evaluation of immune response in BALB/c mice. FMDV type O/IRN/1/2007 was isolated from a cattle in Ray in 2007 and serotyped. The purified VP1 gene was sub-cloned into the PTZ57R/T vector and pcDNA3.1+ expression vector. The PCR product of Vp1 gene without stop codon was sub-cloned upstream of EGFP gene into the pEGFP-N1 vector to evaluate VP1-GFP fusion protein expression. The pcDNA3.1-VP1 and pEGFP-VP1 vectors were transfected into BHKT7 cell line. The expression of VP1 protein was evaluated by SDS-PAGE, western blotting and florescent analysis of VP1-GFP fusion protein. The mice were injected subcutaneously by pcDNA3.1-VP1 vector as DNA vaccine and titration of neutralizing antiserum and T cell proliferation assay were done to evaluate the immune response. Insertion of VP1 gene was confirmed by double digestion of sub-cloned PTZ57R/T, pcDNA3.1+ and pEGFP-N1 vectors. The specific band in western blotting was also confirmed the VP1 protein expression in BHKT7 cells. The expression of VP1-GFP fusion protein was observed under the immune-florescent inverted microscopy as more green florescent spots versus expression of GFP protein, alone. The neutralizing antiserum titer and T cell proliferation increased significantly in the group of mice vaccinated with pcDNA3.1+-VP1 vector verses control groups (P<0.05). The results showed that the target gene was amplified, cloned in the cloning and expression vectors and protein expression was confirmed successfully. According to the confirmed VP1 protein expression and increasing neutralizing antiserum titer and T cell proliferation by pcDNA3.1+-VP1 vector (P<0.05), it can be used as DNA vaccine against FMDV type O/IRN/2007.