Pathobiology Research
Year:2009 | Volume:11 | Issue:3-4|Papers Count:12

Objective: Considering the importance of integrin molecules in the implantation and lack of sufficient information in the expression pattern of these molecules in various phases of estrous cycle. It seemed to be necessary to investigate these molecules in mouse endometrial during the various phases of oestrous cycle. Materials and Methods: Female NMRI mice (n=15) aged 6-8 weeks were studied. Various phases of estrous cycle including: proestrus, estrus, metestrus and diestrus were determined by vaginal smear. The mice were sacrificed (at least 3 per each phase) by cervical dislocation and the tissues were obtained from the middle 1/3 part of their uterine horns at each phase then the cryosections at thicknesses between 8-10 m were obtained. Then the immunohistochemistry were done for integrins of  4, b1, av, b3 and their ligand osteopontine. Results: The integrins were expressed only in the metestrous phase of oestrous cycle in the different locations of mouse endometrium. The positive reactions were observed for av, a4 and b3 in the apical and basal membrane of glandular epithelium. Also the positive reaction for b1 was found in surface and glandular epithelium as well as stroma. The osteopontin expression was seen in the apical membranes of surface and glandular epithelium and was not seen in other locations. Conclusion: It seems that expression of integrins in endometrium is based on their role in the implantation, therefore the molecules a4, b1 and OPN that are expressed on the surface epithelial may be involve in the adhesion of cell to cell and integrins of av, b3 that are expressed in the glandular epithelium and stroma involve in the invasion of embryo.

Objective: The environmental exposure to Magnetic Fields (MFs) may interact with biological systems. MFs are generated from various sources such as power lines, electric appliances at homes and offices, electrified transportation systems including urban railway systems and diagnostic devices such as Magnetic Resonance Imaging (MRI). There are some scientific evidences that imply the exposure to MFs are hazardous to our health and increases the rate of some cancers like leukemia. The biological consequences of exposure to MFs have been investigated from a variety of endpoints. However, most studies have been performed in vitro and have examined effects on cellular processes and its malfunction; such studies can be used as evidence of effects in vivo. Materials and Methods: In this study Bone Marrow Stem Cells were grown in the absence and in the presence of a 15 mT Static Magnetic Field for 5 hours in order to determine any changes in cell cycle progression using the count of cells in different phases. The count of cells in a special phase of cell cycle indicates the length of that phase. The Static Magnetic Field was performed using a locally designed MF generator. Results: A significant increase in the number of cells in G0/G1 was observed in comparison with the controls. Also the number of cells in G0/G1 in the cells treated with Hydrogen-Peroxide, as an oxidative agent, was significantly increased in Static MF. Conclusion: Genetic material damages or mal-function of related proteins may cause these halts. Mfs have not enough energy to affect the biological molecules directly but the mechanism of free radical mediators is probable. These kinds of damages (direct or indirect) can permanently bring the cell cycle to a halt.

Objective: Leishmaniasis is caused by parasitic protozoa of the genus Leishmania which in the infected host is obligate intracellular parasite. LACK gene is conserved among related Leishmania species. LACK is the immuno-dominant antigen of L.major which is considered as the most promising molecule for a recombinant or DNA vaccine against leishmaniasis. Materials and Methods: In this study the genomic DNA of an Iranian standard strain of Leishmania major (MRHO/IR/75/ER) was ertracted and the LACK gene was amplilified by PCR. Then the PCR product was cloned into pTZ57R/T cloning vector, The PT-LACK recombinant plasmid was extracted from transformed E.coli bacteria (TG1 strain) and sequenced. Results: The LACK gene (Accession no LmjF28.2740) of MRHO/IR/75/ER & L. major was amplified using PCR method. LACK gene was cloned into pTZ57R/T coloning vector. Sequence analysis of the cloned LACK gene showed high homology 89% with LmjF28.2740 (LACK gene). Conclusion: The LACK gene of L.major was cloned in pTZ57R/T vector successfully. Recombinant plasmid was confirmed and could be used for the production of recomiomort antigens is DNA vaccines, for further studies.

Objective: Alpha-thalassemia is one of the most prevalent hemoglobin disorders in the world and it is a common hereditary condition caused by deletion of one or more a-globin genes. Common a-thalassemia deletions like 3.7 kb, 4.2 kb, 20.5 kb and Med can be detected by Multiplex PCR. There are, however, some unknown deletions that can not be detected by the mentioned method or even by direct DNA sequencing. In the present study, Real-time PCR was used to determine the presence or absence of unknown deletions. Materials and Methods: Real-time PCR was performed using intercalating dye SYBR Green I and a1, a2 and CLCN7 genes were amplified. Data analysis was conducted using comparative threshold method ( DDCT) for determination of Gene dosage of a1-globin and a2-globin genes. Results: The results showed the ratio of 0.90±0.16 for normal individuals and the ratio of 0.32±0.15 for carrier samples with deletions. In addition, Melting curve analysis confirmed the specific amplification of target genes. Conclusion: The Real-time PCR assay is simple, rapid, and reliable. It can be applied for direct determination of unknown deletions in Alpha-thalassemia carriers.

Objective: Plasmodium falciparum chloroquine resistance is a major problem in malaria endemic areas. Single nucleotide polymorphisms in pfcrt and pfmdr1 genes are known to be associated with chloroquine resistance in some parts of the world. The major goal of the present study was to detect the five single nucleotide polymorphisms in pfmdr1 gene and one single nucleotide polymorphisms in pfcrt gene. Materials and Methods: Total of 26 blood samples were collected from falciparum malaria infectious person with chloroquine failure in Chabahar, a harbor located in Sistan baluchestan during 2 years. Detection of single nucleotide polymorphisms were carried out by Real-Time PCR using Light CyclerTM hybridization probe assay. Results: Our data showed that the pfmdr1 N86Y mutation was detected in 6 (23%) samples. Although this mutation was not observed in the first year but in the second year it was substancial. In addition the pfcrt K76T mutation was detected in 11 samples (42.3%) of CVMNT haplotype, 7 samples (26.9%) of CVIET haplotype, 5 samples (19.2%) of SVMNT haplotype and 2 samples (7.6%) of SVIET haplotype. Conclusion: The mutations considerably have increased during 2 years. Our results showed single nucleotide polymorphisms in pfmdr1 and pfcrt genes. This could be considered as chloroquine resistance markers for malaria control in Chabahar.

Objective: CREB1 is an important downstream protein for many signaling pathways. By designing efficient siRNAs against CREB1, it may be possible to assess the role of molecules involved in signaling pathways in different cell types. In this research the efficiency of CREB1 knockdown by two different siRNAs in K562 cells has been studied. Materials and Methods: siRNAs have been designed according to the criteria suggested by Reynolds et al. K562 cells were transfected by siRNA using Lipofectamine 2000. The efficiency of CREB knockdown has been assessed by quantitative relative Real-time PCR. Results: Our results have shown that only one of the siRNAs has a high level of inhibitory effect on CREB1 gene expression. The expression of CREB1 by this siRNA was knocked-down by 87% in K562 cells. Conclusion: In this research, although two siRNAs were designed according to the Reynolds et al. criteria, only one showed an inhibitory effect. Reasons other than the aforementioned criteria may be involved in effectiveness of siRNAs.

Objective: Herpes simplex virus (HSVs) is a widespread human infectious agent, responsible for persistent and latent infections. Herpes simplex virus infections are usually continually recurrent in the normal population and represent a significant cause of complications in immunocompromised patients. Materials and Methods: In this study HSVs were propagated in BK cells and more than 502 samples were taken and analyzed for HSV IgG antibodies using Virus Neutralization Test (VNT) as golden standard test for evaluating in house Enzyme Linked Immunosorbent Assay (ELISA). Results: Based on the results 80.48 % and 81.67% were positive (1.8) in VNT and ELISA respectively. There was a significant correlation between the VNT and ELISA tests in the tested samples (Pearson’s r = 0.96). Conclusions: Our data showed that the in house ELISA can be used for screening and determination of the prevalence of HSV IgG antibodies, which can facilitates patient management using suitable and cost effective laboratory diagnostic tests.

Objective: Lymnaea palustris was previously found in Mazandaran province but there was not any report about its parasitologic aspects. This study was conducted to finding ecological and Parasitological aspects of snail in Mazandaran province, North of Iran. Materials and Methods: In this descriptive study, more than 181 locations, were checked, in 36 locations, colonies of the snail were found and 490 snails were collected. After diagnosis of snails as Lymanea palustris, in laboratory, they were crushed and their probable cercaria was checked out by a dissecting microscope. Data were analyzed and processed by ArcGIS 9.2 and Microsoft Office 2003 for descriptive analysis. Results: from 490 snails, 6 cases (1.22%) were infected with trematode larval stage. These cercariae were classified as echinostomaercaria. Optimum temperature for the snails was 15-19 degrees of celsius and optimum dissolved salt (TDS) was 200-400 ppm. Population of colonies were raised in autumn and winter but infected snails were seen in summer. Conclusion: This study could show the ecological pattern, distribution, and population dynamic of the snail. Also the existence of echinostomaercaria which is cercaria, generally belong to the Echinostoma sp, indicates veterinary and parasitological importance of local snails. It is probable these parasite, infect man also. More studies on definitive host and exact species of parasite are proposed.

Objective: Toxoplasmosis, caused by an intracellular protozoan parasite, and the Toxoplasma gondii, is widespread throughout the world. In recent years, significant progress has been made in the identification of vaccine candidates which could induce a protective response. GRA7, an excretory 29 kDa Toxoplasma gondii a dense granular antigen released by infected host cells. In tachyzoite-infected cells, p29 accumulates within the parasitophorous vacuole and co-localizes with its delimiting membrane. Materials and Methods: In the present work, first genomic DNA of Toxoplasma gondii was extracted and used for amplifying of GRA7 gene as a template. Then PCR product was extracted from agarose gel and cloned into TOPO vector. The plasmid containing GRA7 gene was extracted from the transformed bacteria (TOP10 strain) and sequenced. Results: Sequence analysis of GRA7 gene cloned into TOPO vector showed only one base difference when composed with the gene bank sequence for RH strain was only one base. Conclusion: The results indicated that this clone is suitable for subcloning in Prokaryotic and Eukaryotic plasmid.

Objective: Occult hepatitis B infection is a form of hepatitis in which despite of absence of detectable HBsAg, HBV-DNA is present in peripheral blood of patients. This clinical form of B hepatitis creates some problems for the Iranian blood transfusion services. Therefore, the aim of this study was the evaluation evaluate of status of occult hepatitis B infection in the Rafsanjanese blood donors. Materials and Methods: In this cross-sectional study, total of 3700 blood donor samples were collected and tested for HBsAg and anti-HBs using ELISA. The HBsAg negative and anti-HBc positive samples were selected and screened for HBV-DNA using PCR. Results: Results of current study indicated that 352 (9.5%) of 3700 blood samples were HBsAg– and anti-HBc+. HBV-DNA was detected in 57 (16.1% of HBsAg–and anti-HBc+ and 1.54% of total samples) samples. Conclusion: Results of this study are in agreement with our previous studies in the prevalence of OBI. Therefore, it seems that occult hepatitis B infection rate is high in the Iranian blood donors and probably is one of the main causes of post-transfusion hepatitis.

Objective: Dendritic cells (DCs) play a critical role in the beginning and in the course of immune responses. By observing. Considering the defect in these cells in patients with tumor malignancy. In recent years there has been considerable interests in correction and use of these cells. In our prvious studies we showed that lysate of Listeria monocytogenese can induce maturation of dendritic cells, and induce TH1 response and increase the survival of tumor in an experimental model. The main objective in the present study is to evaluate the effects of different constituents of Listeria monocytogenes on DCs and their efficiency to induce TH1 response. Materials and Methods: After preparation of different components of Listeria monocytogenese (lysate of Listeria monocytogenes, protein and nucleic acid components), mouse bone marrow cells were cultured for 5 days in the presence of IL-4 and GM-CSF and treated with different components of Listeria for another 2 days. DCs were evaluated for the expression of Co-stimulatory molecules, MHC Class II molecules and Cytokine secretion. Results: The results showed that, all of the components are able to mature DCs efficiently and induce TH1 response. But dendritic cells matured with protein components shift the response to TH1 more efficiently. Conclusion: Our findings indicated that dendritic cell maturation protein components of Listeria monocytogenese shift the response to TH1 more efficiently and may have beneficial effects in cancer immunotherapy.