Iranian Journal of Basic Medical Sciences
Year:2014 | Volume:17 | Issue:7|Papers Count:12

Objective (s): Genetic analysis of two consanguineous Pakistani families with localized autosomal recessive hypotrichosis was performed with the goal to establish genotype-phenotype correlation.Materials and Methods: Genomic DNA extraction had been done from peripheral blood samples. Extracted DNA was then subjected to PCR (polymerase chain reaction) for amplification. Linkage analysis was performed using 8% polyacrylamide gel. Candidate gene was sequenced after gene linkage supported at highly polymorphic microsatellite markers of the diseased region.Results: Both families were initially tested for linkage to known genes, which were involved in human hereditary hypotrichosis, by genotyping Highly polymorphic microsatellite markers. Family B showed partial linkage at P2RY5 gene on chromosome 13q14.11-q21.32; hence, all exonic regions and their introns boundaries were subjected to DNA sequencing for any pathogenic mutation.Conclusion: Both families were tested for linkage by genotyping polymorphic microsatellite markers linked to known alopecia loci. Family A excluded all known diseased regions that is suggestive of some novel chromosomal disorder. However, sequencing of P2RY5 gene in family B showed no pathogenic mutation.

Objective(s): Hypothermia and decompressive craniectomy (DC) have been shown to be neuroprotective. This study was designed to evaluate neuroprotective effects of delayed singular or combination of DC and local hypothermia on stroke.Materials and Methods: Cerebral ischemia was induced in 48 Wistar rats assigned to 4 groups: control, decompressive craniectomy (DC), local hypothermia (LH), combination of hypothermia and craniectomy (HC). Infarct size and BBB disruption were measured 48 hr after ischemia insult. Neurological deficits were assessed at 24 and 48 hr after stroke by using sticky tape test, hanging-wire test and Bederson’s scoring system. BBB disruption was measured by Evans blue dye leakage.Results: Although infarct size was significantly reduced in LH, DC and HC groups (P<0.001), combination therapy was more neuroprotective compared to craniectomy alone (P <0.01). BBB disruption was significantly reduced in DC (P <0.05) and LH and HC (P <0.01).While sticky tape test (P <0.05 at 24 hr; P <0.001 at 48 hr) and hanging-wire test (P <0.05) showed better behavioral performance only in HC, Bederson test showed improved behavioral functions of both LH (P <0.05 at 24 hr and P <0.01 at 48 hr) and HC animals (P <0.01). Neurological deficits were also decreased in LH (P <0.05) or HC (P <0.05 at 24 hr; P <0.01 at 48 hr) groups compared to the DC group at the same time.Conclusion: Based on our data, although both delayed local hypothermia and craniectomy are protective after stoke, combination therapy of them is more neuroprotective than given alone.

Objective (s): The aim of this study was to evaluate distribution and changes of glycoconjugates of retinal photoreceptors during both pre- and postnatal development.Materials and Methods: Tissue sections from days 15 to 20 of Wistar rat embryos and 1 to 12 postnatal days of rat newborns including developing eye were prepared for lectinhistochemistry technique. Horseradish peroxidase (HRP) -labeled lectins including Vicia villosa (VVA), peanut agglutinin (PNA), Maclura pomifera (MPA) and wheat germ agglutinin (WGA-ІІ) were used. Alcian blue (pH 2.5) was used for counterstaining.Results: Interphotoreceptor matrix (IPM) plays a crucial role in photoreceptors differentiation and acts as a mediator in interactions between photoreceptors and retinal pigment epithelium (RPE). Specific cell surface glycoconjugates secreted from cone cells could help us to distinguish these cells from rod photoreceptors. Our results for the first time revealed the strong reaction of cone photoreceptors with the cone-specific lectin (PNA) at postnatal day 12 (P12). Postnatal day 12 can be determined as the final differentiation of cone photoreceptors.Conclusion: According to our findings, we suggest that the generation of the eye photoreceptors begins from pre- natal period and their final differentiations will continue to postnatal period. Glycoconjugates including (b-D-Gal [1–3] -D-GalNac) and (b-D-Gal) terminal sugars play a critical role in the pre- and postnatal development and differentiation of retinal photoreceptors.

Objective (s): OCT4 is a transcription factor required for pluripotency during early embryogenesis and the maintenance of identity of embryonic stem cells and pluripotent cells. Therefore, the effective expression regulation of this gene is highly critical. UTR regions are of great significance to gene regulation. In this study, we aimed to investigate the potential regulatory role played by 5’UTR and 3’UTR of the Oct4 gene in mouse BMSC and P19 cells.Materials and Methods: The Oct4 5’UTR and 3’UTR sequences were cloned into pGL3 luciferase plasmid which led to the generation of pGL3 5’-UTR, pGL3 5’& 3’-UTRs and pGL3 3’-UTR vectors. The vectors were transfected into BMSC and P19 cells followed by luciferase assay.Results: The assay of luciferase expression exhibited a direct link between the presence of Oct4 3’- UTR and the decrease of luciferase count in both cell lines; whereas 5’UTR indicated diverse behaviors in two cells. This discrepancy could be explained in view of the difference of cellular contexts in which the Oct4 UTRs act.Conclusion: This study sheds some light on the role of UTR regions of mouse Oct4 in regulating post-transcriptional gene expression in pluripotent cells. These data represent potential to be used for the development of novel therapeutic approaches for a variety of malignancies.

Objective (s): Prostate cancer is the world’s leading cause of cancer and the second cause of cancer-related death in men after lung cancer. Differentiation of prostate adenocarcinoma from benign prostate lesions and hyperplasia sometimes cannot be done on the basis of morphologic findings. Considering the fact that in the prostate adenocarcinoma there is no basal cell layer, basal cell markers can help to differentiate prostate adenocarcinoma from cancer mimickers.Materials and Methods: We studied 98 prostate biopsy blocks (40 adenocarcinoma and 58 benign lesions) for basal cell marker expression.Results: p63 and 34bE12 were negative in all prostate adenocarcinoma specimens, but all benign prostate hyperplasia and high grade intraepithelial neoplasia cases expressed them.Conclusion: Basal cell markers can help to distinguish prostate adenocarcinoma from cancer mimickers.

Objective (s): According to the prevalence of neoplastic diseases, there is a deep necessity for discovery of novel anticancer drugs in the field of medicinal chemistry. In the current study, a new series of phenylthiazole derivatives (compounds 4a-4f) was synthesized and their anticancer activity was assessed in vitro.Materials and Methods: All synthesized derivatives were evaluated towards three human cancerous cell lines of SKNMC (Neuroblastoma), Hep-G2 (Human hepatocarcinoma) and MCF-7 cell (Breast cancer) using MTT assay and obtained values (IC50±SD) were compared with doxorubicin.Results: Unfortunately, none of the synthesized compounds showed superior activity than doxorubicin against cancerous cell lines. MCF-7 cell line was the most resistant cell line against tested compounds. Compounds 4c with para nitro (IC50=10.8±0.08 mM) and 4d with meta chlorine (IC50=11.6±0.12 mM) moieties exerted the highest cytotoxic effects towards SKNMC and Hep-G2 cell lines respectively.Conclusion: A new series of phenylthiazole derivatives were synthesized and their anticancer activity was assessed against cancerous cell lines. More structural modifications and derivatization is necessary to achieve to the more potent compounds.

Objective (s): Experimental allergic encephalomyelitis (EAE) is an autoimmune disease validated as animal model of multiple sclerosis (MS). Administration of genistein, a phytoestrogenic component of soy, to mice at the onset of EAE is known to attenuate the clinical signs of the disease. The potential effects of genistein on established EAE is less studied. In the current study, we aimed to compare the effects of genistein administration on EAE severity in early and late phases of the disease.Materials and Methods: The C57BL/6 mice were induced with EAE, using MOG 35-55 and gavaged with genistein (300 mg/kg) either after the appearance of the first clinical sign or 30 days post disease induction for ten days.24 hr after the last gavage, mice were sacrificed. Brains and spleens were removed for assessing lymphocyte proliferation, cell cytotoxicity, and cytokine profile. Spinal cords were dissected to assess the amount of demyelination using Luxol fast blue/cresyl violet staining.Results: Administering mice with genistein, after the establishment of EAE, did not reverse the clinical signs of disease. However, treating with genistein at the onset of disease alleviated the clinical signs by reducing neuronal demyelination. Genistein suppressed the production of IFN-g and enhanced IL-10 secretion in splenocyte and brain. Genistein also reduced IL-12 and TNF-a secretion in splenocytes, suppressed the proliferation of T-cells, and reduced the cell cytotoxicity.Conclusion: Genistein oral therapy might only reduce EAE severity if started in early phases of the disease.

Objective (s): Several studies have been carried out to investigate the effect of various nanoparticles exposed to radiofrequency (RF) waves on cancerous tissues. In this study, a colon carcinoma tumor model was irradiated by RF in the presence of gold-gold sulfide (GGS) nanoshells.Materials and Methods: Synthesis and characterization of GGS nanoshells were initially performed. CT26 cells were subcutaneously injected into the flank of BALB/c mice to create the colon carcinoma tumor models. Then the tumors were subjected to different treatments. Treatment factors included intratumoral injection of GGS and RF radiation. Different groups were considered as control with no treatment, receiving GGS, RF irradiated and simultaneous administration of GGS and RF. Efficacy of the treatments was evaluated by daily monitoring of tumor volume and recording the relative changes in it, the time needed for a 5-fold increase in the volume of tumor (T5) and utilizing pathologic studies to determine the lost volume of the tumors.Results: In comparison with control group, tumor growth was not markedly inhibited in the groups receiving only GGS or RF, while in the group receiving GGS and RF, tumor growth was effectively inhibited compared with the other groups. In addition, the lost volume of the tumor and T5 was markedly higher in groups receiving GGS and RF compared with other groups.Conclusion: This study showed that RF radiation can markedly reduce the tumor growth in presence of GGS. Hence, it can be predicted that GGS nanoshells convert sub-lethal effects of noninvasive RF fields into lethal damages.

Objective (s): The present study was undertaken to determine the effects of acute exposure to malathion on oxidative stress and cytotoxic potential of anticholinesterase insecticide, malathion in reproductive toxicology of adolescent male mice.Materials and Methods: Thirty two adolescent male mice at pubertal age were treated with 500 mg/kg body weight (BW) of malathion for three days. After exposure, biochemical markers and sperm analysis were evaluated and finally histological modifications of testis and sperm were assessed.Results: Our data showed that treatment of male mice with malathion (500 mg/kg, BW) could lead to oxidative stress. Induced oxidative stress status can be assessed due to increased malondialdhyde (MDA) content, decreased thiol group content, as well as increased antioxidant enzyme activities. On the other hand, exposure to malathion at the pubertal age led to alteration of semen parameters; sperm production and percentage of motile sperm were decreased in the treated groups compared to the control. Furthermore, exposure of male mice to malathion led to a decrease of testosterone level, inhibition of acetyl cholinesterase, and decrease of the reproductive performance of male mice after three days of treatment at the age of puberty.Conclusion: The importance to carry out in vitro reproductive toxicology assays lies on the need of knowing the alterations these insecticides may cause at cellular level, since they are endocrine disruptors that interfere with reproductive functions.

Objective(s): HTLV-1 is the first human retrovirus that has been recognized and is associated with HAM/TSP and ATLL. Studies have shown that less than five percent of HTLV-1 infected carriers develop HAM/TSP or ATLL and about ninety-five percent remain asymptomatic. Therefore, the proviral load with Tax may affect cellular genes such as cytokines and oncogenes, as well as involve in pathogenicity.Materials and Methods: Thirty HAM/TSP patients, thirty HTLV-1 healthy carriers, and MT-2 cell line were evaluated for HTLV-1 activity. PBMCs were isolated and activated using PMA and ionomycine. Real-time PCR and TaqMan methods were performed using specific primers and fluorescence probes for Tax expression and proviral load assessment. Β2microglobulin (b2m) and albumin were used as controls in Tax expression and in proviral load, respectively.Results: An insignificant increase in Tax expression was observed in rest PBMCs of HAM/TSP patients compared to healthy carriers. However, after lymphocyte activation there was a significant increase in Tax expression in HAM/TSP patients (P=0.042). The Proviral load in patients was significantly higher than in carriers. Moreover, there was a significant correlation between Tax mRNA expression in activated PBMCs and proviral load (R=0.37, P=0.012).Conclusion: Although proviral load had been addressed as a valuable index for monitoring HTLV-1 infected subjects, the results of this study demonstrated that Tax expression in activated PBMCs along with proviral load assessment in HAM/TSP patients are a more reliable factor for determining the prognosis and monitoring healthy carriers and HAM/TSP patients.

Objective (s): Various species of genus Trigonella are important from medical and culinary points of view. The essential oil of Trigonella torbatejamensis Ranjbar as an endemic plant in Iran has not been studied previously. The essential oil of this plant was analyzed by different methods for identification of its components.Materials and Methods: The essential oil composition of aerial parts of T. torbatjamensis was obtained by hydro-distillation and analyzed by GC-FID and GC-MS apparatus.Results: Forty components, representing 98.5% of the total components, were identified. The pattern of the main grouped components in essential oil was: sesquiterpene hydrocarbons (70.2%), oxygenated sesquitepenes (16.5%), oxygenated monoterpenes (3.5%) and monoterpene hydrocarbons (0.5%). Germacrene –D (33.0%), bicyclogermacrene (26.0%), and viridiflorol (5.3%) were the main components of the essential oil.Conclusion: The essential oil of T. torbatjamensis Ranjbar consisted of forty components with sesquiterpene hydrocarbons as the main group of components.

Objective (s): To investigate the effect of Eucheuma cottonii on alveolar macrophages (AM) and malondialdehyde (MDA) levels in bronchoalveolar lavage fluids (BALF) in particulate matter 10 (PM10) coal dust-exposed rats.Materials and Methods: Ten groups, including a non exposed group and groups exposed to coal dust at doses of 6.25 (CD6.25), 12.5 (CD12.5), or 25 mg/m3 (CD25) an hour daily for 6 months with or without supplementation of ethanolic extract of E. cottonii at doses of 150 (EC150) or 300 mg/kg BW (EC300). The number of macrophages was determined using a light microscope. MDA levels were measured by TBARS assay.Results: EC150 insignificantly (P>0.05) reduces the AM in CD groups compared to non treatment groups. EC150 and EC300 significantly (P<0.05) decreased MDA levels in CD12.5 and CD25 groups relative to non treatment groups.Conclusion: E. cottoniiattenuated oxidative stress in chronic exposure of PM10 coal dust.