Journal of Pathobiology Reaearch
Volume:15 Issue: 4, 2013

Platelets are anucleated fragments derived from megakaryocytes. It has been demonstrated that platelets play a role in hemostasis and innate immunity. In addition, platelets have a CD40 ligand which is an important molecular marker in motivating immune cells. Thus, platelets also have a role in adaptive immunity as seen by their ability to activate B cells. Since human platelet microparticles (MPs) originate from platelets, we have chosen to examine the effects of MPs on B cell activation. Platelet MPs were isolated from platelet concentrates obtained from theTehranBloodTransfusionCenter. The MPs were co-cultured with B cells isolated from human whole blood with magnetic beads using negative selection. After seven days, the expression of activation markers CD27 and CD86, as well as IgD were evaluated by flow cytometry. In a comparison between test (B cells/MPs) and control (B cells) cells we observed that the expression of activation markers CD27 and CD86 increased during the seven-day co-culture period. However, the expression of IgD antibody decreased. As with platelets, MPs can affect B cell activation during in vitro co-culture.

Electro-magnetic radiation (EMR) is emitted from mobile phones. Various researches have shown relationships between mobile phone EMR exposure to cancer and neurologic damages. This study aims to investigate the effects of mobile phone EMR on brain antioxidant enzyme activity and the learning process. Rat pups and their dames were exposed to EMR for 3 h per day from P2 to P14. After separation of male and female rats on P22, the rats were housed in an air room under normal animal conditions. From P59 to P61, male rats were trained three times per day for a total of 3 days. On P62, their behavior was assessed. The rats were sacrificed by decapitation and the levels of superoxide dismutase and catalase in their brains were assessed. The amount of time to locate the hidden platform and time spent exhibiting freezing behavior increased in exposed group compared to the control group. Superoxide dismutase and catalase activities were reduced in the mobile phone group. Additional studies are necessary to clarify the relationship between mobile phone radiation and brain tissue with regards to antioxidant enzyme activities, learning and memory. Our results suggest that mobile phone radiation may lead to decreased learning that is induced by abnormalities in antioxidant enzyme activities.

This study presents a simple method for isolation, expansion and purification of neonatal mouse spermatogonial stem cells. We used enzymatic digestion to isolate a cell suspension of spermatogonia and Sertoli cells from neonatal 2-day-old mice. The cells were cultured in DMEM/F12 that contained 10% serum for two weeks. Sertoli and spermatogonia cell characteristics were confirmed by examining for the presence of vimentin and PLZF proteins, respectively. To assess the rate of spermatogonia stem cell expansion, the area and number of colonies were measured during the two weeks of culture. At the end of the second week, we detected spermatogonia cell-specific expressions of the Stra8, Piwill2, DAZL, and Mvh genes. Current results indicated that isolated Sertoli and spermatogonia cells were immunopositive for specific markers. During the culture period, a significant difference was seen in the number and area of spermatogonial stem cell colonies (P<0.05) at four time points. In addition, spermatogonial specific gene expression demonstrated that these cells were undifferentiated after two weeks of culture. Our study showed that co-culture of spermatogonia and Sertoli cells from same source provides a convenient and efficient environment. This co-culture, without the addition of external growth factors and chemical manipulations, can be used for proliferation of spermatogonia stem cells.

Astrocytes are the most abundant glial cell type. They may promote or inhibit CNS inflammation depending on which cytokines are secreted. Astrocytes also have immune roles. IL-19, IL-20, and IL-24 activate a heterodimer receptor composed of the IL-20R1 α-chain and the IL-20R2 β-chain. It has long been considered that signaling by these receptor complexes affects immunological reactions, however the biological functions of IL-20R1 and IL-20R2 in the brain remain unclear. As the first step to address the role of these cytokine receptors in the brain, in this study we have researched the expressions of IL-20R1 and IL-20R2 in C57BL/6 mice astrocytes. We examined expressions of IL-20R1 and IL-20R2 proteins in mice astroglial cells and in the 1321N1 astrocytoma cell line in response to MOG, LPS and GM-CSF by flow cytometry. The effect of LPS on mRNA expression of IL-20R1 and IL-20R2 was investigated by RT-PCR. We provide, for the first time, evidence that astrocytes expressed IL-20R1 and IL-20R2 mRNA not only in response to LPS stimulation but also in unstimulated astrocytes. We did not observe the expressions of IL-20R1 and IL-20R2 proteins in mice astroglial cells and the 1321N1 astrocytoma cell line. IL-20R1 and IL-20R2 mRNA are constitutively expressed in astrocytes. Because the majority of neuropathological processes involve astrocytes and inflammatory cytokines, the results of this study, which are reported for the first time, have important implications for future research.

the aim of this study was to evaluate the effect of sucrose on follicular survival rate and the incidence of apoptosis in rat ovarian tissue following vitrification.

Ovaries of approximately 5-week-old female Wistar rats were divided randomly into three groups: control (non-vitrified), VI (Ethylene Glycol + Dimethyl Sulfoxide) and VII (EG + DMSO + 0.25 mol/lit sucrose). Vitrified-warmed samples were incubated for approximately 30 minutes and fixed in Bouin’s fixative. The samples were serially sectioned and stained either with H&E or immunohistochemistry kit of anti-active and a pro-caspase-3 kit.

Data analysis showed that the rate of growing follicles that survived, with the exception of primordial follicles, was comparable between the vitrified-warmed and control samples. Morphologically healthy primordial follicles showed significant reductions in all vitrification groups compared to the control, however this rate was not significant between the vitrification groups. In comparison with healthy follicles, there were significantly more dead follicles in the vitrification groups than the control group. In addition the apoptotic follicles increased significantly after vitrification, with the exception of the antral follicles. Although the number of apoptotic follicles was similar between both vitrification groups, however there were significantly more preantral apoptotic follicles in the VII group compared to the VI and control groups.

According to these results, the presence or absence of sucrose has no significant effect on the preservation of primordial and primary follicles which are important for transplantation.

Common variable immunodeficiency (CVID) is one of the most frequent cases of primary immunodeficiency. It is likely that this heterogeneous disease is caused by several distinct genetic disorders. The activation-induced cytidine deaminase (AID) enzyme is involved in class switching, somatic hypermutation (SHM) and processes associated with gene conversion in the germinal center. In order to clarify the possible role of AID in the pathogenesis of CVID, we have studied the AID gene expression in CVID patients. Peripheral blood mononuclear cells (PBMC) from 21 patients and healthy controls were isolated. The isolated cells were stimulated by CD40L and IL-4 to induce AID gene expression. After five days, total RNA from the stimulated cells was extracted and AID gene expression was investigated by RT-PCR. RT-PCR results showed that after stimulation by CD-40L and IL-4, the AID gene was expressed in all of the samples. The control samples were also positive for AID gene mRNA expression. In this investigation we studied the expression of AID gene in CVID patient's B lymphocytes for the first time. Regards to our results which showed that all patients normally expressed the AID gene mRNA and considering that one of the main problems in a number of CVID patients is disorders in phenomena related to the germinal center and complete differentiation of B lymphocytes, it can be concluded that possible defects in other molecules involved in class switching is responsible for this disease. Understanding the various genetic defects responsible for this heterogeneous disease could lead to its division into more homogenous subtypes with distinct therapeutic strategies, so further investigations is recommended.

Breast cancer is the second leading cause of cancer death in women. Cisplatin is a traditional cancer drug commonly used in chemotherapy for killing cancer cells. Modulation at the mRNA levels of apoptotic related genes often correlate with the sensitivity of various types of cancer cells to chemotherapeutic agents. Nanoparticulate drug delivery systems are being developed to effectively deliver smaller doses of chemotherapeutic agents and control drug distribution in the body. In this study, we evaluate the expressions of BCL2 and BAX genes in T47D treated with cisplatin and cisplatin nanoparticles, which can result in a new approach to breast cancer therapy. In this study, we treated T47D cells with different concentrations of cisplatin and cisplatin nanoparticles at 48 h. The IC50 was determined. We extracted RNA by using RNX solution, after which cDNA was synthesized. The precise primers for the BCL2, BAX and TBP genes were designed by specific software. The quantity of BCL2 and BAX gene expression compared to TBP gene (reference gene) was analyzed using real-time PCR. BCL2 and BAX gene expression levels in T47D cells treated by cisplatin were 0.7 (BCL2) and 1.48 (BAX); in T47D cells treated with cisplatin-loaded nanoparticles, the gene expressions were 0.03 (BCL2) and 2.41 (BAX). In this study, the results have shown that cisplatin-loaded nanoparticles are effective anticancer agents. Cisplatin nanoparticles induce apoptosis in human breast cancer cell lines. We have shown that cisplatin nanoparticles strongly increased cytotoxicity in comparison to the free drug in the T47D cell line.

Different signaling pathways have been identified that are involved in the cellular response to opiates. The mitogen-activated protein kinase pathway is one of the most important signaling pathways underlying the neuronal response to opiates. MicroRNAs (miRNAs) are considered to be post-transcriptional regulators of gene expression with paramount significance, which plays key roles in modulating cellular processes such as neuronal plasticity and synaptic consolidation. The purpose of this study is to identify miRNAs that are differentially expressed in response to chronic morphine treatment, and predict those genes that have a possible role in this process. Because the MAPK pathway in involved in morphine dependence and participates in hypersensitivity to pain, determining miRNAs that modulate this pathway could be insightful in morphine dependence treatment and pain control.

In this study, the BE(2)-C neuroblastoma cell line was chronically treated with morphine sulphate and the changes in expression of 750 miRNAs were analyzed by real time PCR.

Two up- and down- regulated groups of miRNAs were determined to be differentially expressed in response to morphine: i) has-mir-193a-3p, -212, -181c, -362-3p, -639, -646 and ii) has-mir-412, -937, -558, -552, -943, -628-5p, -593, -555, -636, -643, 566, -571, -642, -653, -611, -31, let7-g.

The analysis of differentially expressed miRNAs showed that the MAPK signaling pathway could be regarded as a signaling pathway with utmost significance in chronic morphine response. Due to the role played by MAPK pathway in cellular response to morphine exposure, we can propose that protein phosphorylation has a presumable part in this response.

The aim of the this study was to investigate the prevalence of toxB, paa, lpf and iha adhesion genes in enteropathogenic Escherichia coli (EPEC) isolates lacking in two important adhesion factors, the eaeA and bfpA genes. We examined a total of 70 serologically confirmed EPEC (eaeA-, bfpA-) isolates. DNA from the isolates was extracted by the phenol-chloroform method. toxB, paa, lpf and iha adhesion genes in the EPEC isolates were detected by polymerase chain reaction. Data were analyzed by SPSS software and statistical analysis using the chi square test. P-values less than 0.05 were considered significant. PCR was positive for the toxB gene in 2 (2.85%), paa in 3 (4.28%), lpf in 32 (45.71%) and iha in 15 (21.42%) of the 70 strains. Statistically, none of the toxB, paa, and lpf genes were associated with diarrhea. However, the iha gene showed a weak significant relation to diarrhea (P=0.11). The main mechanism of pathogenicity for EPEC is attachment and effacement. Therefore, EPEC (eaeA-, bfpA-) should have another adhesin factor, which should be investigated. EPEC strains (eaeA-, bfpA-) that possess the lpf gene are common. Further investigations of the virulence properties of these strains are necessary in order to elucidate the role of these virulence factors in diarrhea among Iranian children.