Iranian Biomedical Journal
Volume:13 Issue: 2, Apr 2009

Axotomy causes sensory neuronal loss. Reconnection of proximal and distal nerve ends by surgical repair improves neuronal survival. It is important to know the morphology of primary sensory neurons after the surgical repair of their peripheral processes. Animals (male Wistar rats) were exposed to models of sciatic nerve transection, direct epineurial suture repair of sciatic nerve, autograft repair of sciatic nerve, and sham operated. After 1 and 12 weeks of the surgery, the number of L5 dorsal root ganglion (DRG) and ultrastructure of L4-L5 DRG neurons was evaluated by fluorescence and electron microscopy, respectively. Nerve transection caused sensory neuronal loss and direct epineurial suture but no autograft repair method decreased it. Evaluation of morphology of the neurons showed classic features of apoptosis as well as destructive changes of cytoplasmic organelles such as mitochondria, rough endoplasmic reticulum and Golgi apparatus in primary sensory neurons. These nuclear and cytoplasmic changes in primary sensory neurons were observed after the surgical nerve repair too. The present study implies that the following peripheral nerve transection apoptosis as well as cytoplasmic cell death contributes to neuronal cell death and reconnection of proximal and distal nerve ends dose not prevent these processes. Iran. Biomed. J. 13 (2): 65-72, 2009

The aim of this study was to assess effects of cysteamine as an anti-oxidant on rate of in vitro maturation of oocyte and determination of its effects on spindle size and shape. Pre-mature mice were primed with pregnant mare stimulating gonadotrophin (PMSG) and germinal vesicle (GV) stage oocytes were obtained 48 h after. The oocytes were cultured in tissue culture medium (TCM 199) with 50, 100, 200 and 500 µM cysteamine. Experiments also included a group of ovulated oocytes (in vivo matured) after priming with PMSG and human chorionic gonadotrophin. For spindle immuno-cytochemistry of metaphase II (MII), oocytes α and β tubulin antibody were performed. Chromosome staining was performed with Hoechst. Our results showed that the rate of GV breakdown (GVBD) and MII increased in all cysteamine groups except in group cultured with 500 µM cysteamine. Immuno-cytochemistry analysis showed that spindle area in all in vitro Maturation oocytes increased when compared to in vivo oocytes. Spindle shape and size (spindle area) in 100 µM cysteamine in comparison to in vivo group was insignificant (P>0.05). Our results revealed that cysteamine improved IVM rate in a dose dependant manner. The size and shape of spindle in the presence of given concentration of cysteamine was similar to in vivo. Therefore, cysteamine improved microtubule organization, rate of GVBD and MII development. Iran.

Pharmacogenomics is the study of genetic variations among individuals to predict the probability that a patient will respond to single or multidrug chemotherapy. Breast cancer is one of the most common cancers among women worldwide. Treatment of breast cancer by application of biological rationales gives us the ability to match the correct pharmacology to individual tumour genetic profiles. The breast cancers exhibit multiple anomalies in phosphatidylinositol 3 kinase pathways, such as phosphatase and tensin homolog deleted on chromosome TEN loss that can be put in context of therapy with rapamycin analogues. Considering the high incidence of breast cancer in Iran, the potential role of tumor suppressor PTEN/MMAC1gene was investigated in Isfahanian breast cancer patients. In this study, PTEN was evaluated by means of polymerase chain reaction, single strand conformation polymorphism, Heteroduplex mobility assay and direct DNA sequencing in 72 breast cancer tumors for detection and characterization of mutations. According to the results of this research, nucleotide substitutions were found in 5/72 (7%) of samples. The sporadic breast cancer patient was found to be heterozygote for the p.D92N, p.C105W, p.D107N, p.A121P and p.R130Q mutations. One novel mutation, p.D107N, was found in this study. Loss of PTEN function in breast cancer can occur either by mutation or reduction of PTEN expression in almost half of sporadic breast tumors. This rate of mutations is an important consideration for novel therapeutic in which biological efficacy is influenced by the activity of PTEN

The present study was designed to evaluate the homing potential of mouse embryonic stem cells (ESC) treated with erythropoietin (EPO) in hematopoietic organs such as spleen and liver after transplantation using morphological and immuno-histochemical techniques. Day-four embryoid body (EB)-derived cells were dissociated and re-plated in medium in the presence and absence of EPO for three days. The EPO- and untreated differentiated cells were labeled with 5-bromo-2 deoxyuridine (BrdU) before transplantation and analyzed using flow cytometry and reverse transcription-PCR methods. BrdU-labeled cells were injected via the tail vein into irradiated adult mice in both groups. The spleen colony-forming unit assay (CFU-S) was performed 12 days after transplantation. Immuno-histochemistry was also carried out to trace transplanted cells. The percentage of CD34 positive cells was 5.51 ± 1.06% in the EPO-treated group and 1.63 ± 0.225% in untreated group. The RT-PCR analysis showed that the EPO-treated cells expressed ε globin, βH1 globin, RUNX1 and EPO receptor genes, but the beta-major globin gene was not expressed. The number of colonies formed in the spleens of treated group (17.33 ± 4.726) was significantly different from the control group (6 ± 1). The population of BrdU positive cells in spleen of EPO-treated cell-transplanted group was higher than that of the control group. Also, BrdU positive cells were observed in the central vein of the liver sections of EPO-treated and control groups but were not observed in the liver parenchyma. There were not BrdU positive cells in the spleen and liver sections of the sham group. Our results confirm that ESC have the ability to home and form colonies in spleen after transplantation and EPO-treated EB-derived cells caused an increase in the number of colonies in spleen after CFU-S

Endometrial remodeling occurs during each menstrual cycle in women. Reports have shown that, in a variety of cell types, processes such as proliferation, signaling complex formation and extra cellular matrix remodeling require a cytoplasmic tyrosine kinase, focal adhesion kinase (FAK). The present study has focused on the expression pattern of FAK in human endometrium during the menstrual cycle. The purpose of this study was to ascertain the probable function of FAK in menstrual cycle changes and the role of FAK in tissue repair and tissue remodeling in vivo. Formalin-fixed paraffin-embedded endometrial samples were obtained from 400 pre-menopausal, non-pregnant women, who underwent hysterectomy and biopsy for benign diseases. Forty six samples with no tissue abnormalities were studied and ABC staining method of immuno-histochemistry methods was applied. Positive staining of FAK by different cell types of human endometrium was scaled and compared with each other by using histologic score method. All different cell types of endometrium showed various patterns of FAK expression in different stages of menstruation. FAK in glandular and luminal epithelial cells is up-regulated during the early proliferative (EP) to mid-secretory (MS) phases. FAK in stromal cells is up-regulated during the EP, early and MS phases in comparison to the late secretory (LS) phase. FAK expression in endothelial cells is up-regulated during the EP and MS phases in comparison to LS phase. This study showed that endometrial FAK expression is a phase-dependent manner during the menstrual cycle. It appears that up-regulation of FAK during the proliferative phases is responsible for endometrial regeneration and high expression of FAK in the EP and MS phases may associate with the implantation. Down-regulation of FAK during the LS phase may facilitate apoptosis in human endometrium. It seems that FAK as a key kinase plays a critical role in endometrial remodeling that it may regulate by steroid hormones

Cholesteryl ester transfer protein (CETP) plays a main role in high-density lipoprotein metabolism. CETP gene possesses several single nucleotide polymorphisms which have been associated with plasma high-density lipoprotein cholesterol (HDL-C) concentrations. The aim of this study was to determine the association of CETP -629C/A and I405V polymorphisms with coronary artery disease (CAD) in Iranian population. The presence of two CETP gene polymorphisms -629C/A and I405V were studied in 187 unrelated CAD cases and 136 controls. All the samples were clinically examined and lipid profile was estimated. Genotyping was performed using polymerase chain reaction/restriction fragment length polymorphism method. The frequency of -629C/A and I405V allelic variants were found to be 0.732 and 0.366 in cases and 0.658 and 0.348 in controls, respectively. The frequency of A allele of -629C/A polymorphism in cases was significantly higher than that of controls. HDL-C in AA genotype was higher than CA and CC genotypes in controls. No significant effect of II, IV and VV genotypes was found in lipid profiles. No significant association was found between CETP I405V polymorphism and increased risk of CAD in Azeri population studied. AA genotype of -629C/A increased HDL but the risk of CAD in this genotype might be higher than CC genotype

This study was undertaken to establish the characterization of cultured oral mucosal epithelium and introducing them as an alternative source for reconstruction of ocular surface disease. Human oral epithelial cells were cultured on simple media (DMEM/HF12) as control and co-cultured on mitomycin C-treated 3T3 feeder layer, on the amniotic membrane (AM) without nitrocellulose and the mitotically inactivated 3T3 fibroblast, and on the sandwich layer of AM fastened on the nitrocellulose as insert and 3T3 fibroblast. After 3 weeks, the characteristics of the cells were assessed morphologically and also ultrastructurally using scanning electron microscopy and transmission electron microscopy and immuno-cytochemically. The epithelial cells were cultured on AM spread on nitrocellulose insert and 3T3 feeder layer showed better growth than other groups and all groups of study were shown similar characteristics. The cultured oral epithelial shared the characteristics with corneal epithelium. Thus the oral epithelial could be an alternative source for transplantation

Bone marrow stromal cells (BMSC) are used as a source for cell therapy in different model for neurological disorder such as stroke and spinal cord injury. However, the transdifferentiation of BMSC into cholinergic phenotype requires more investigation. BMSC were isolated from adult rats, pre-induced with β-mercaptoethanol (BME) and followed by nerve growth factor (NGF) induction. Neurofilaments of 68 kDa, 160 kDa and 200 kDa (NF-200, NF-160 and NF-68, respectively) immuno-staining were used for evaluating the transdifferentiation of BMSC into neuronal phenotype. The percentage of neurofilaments immuno-reactive cells was applied in order to evaluate the results at the pre-induction and the induction stages. Also, NeuroD and Oct-4 expressions, using RT-PCR, were used in assessing the progression of BMSC into neuronal lineage. Choline acetyltransferase immuno-reactive cells were used for estimating the percentage of cholinergic neuronal phenotype. Immuno-staining with anti-microtubule-associated protein-2 (MAP-2) and anti-synapsin-I antibodies was done in order to evaluate cell tendency for synaptogenesis. The yield of cholinergic neurons with BME as pre-inducer and NGF as inducer was 80%. Also, NF-200, NF-160, NF-68, MAP-2 and synapsin-I were detected in the transdifferentiated cells. RT-PCR showed the expression of NeuroD, while Oct-4 was not detected. BME as pre-inducer and NGF as inducer for BMSC transdifferentiation into cholinergic phenotype are potential sources in traumatic injury therapy in the central nervous system