An Investigation of the Retinal Layers in an Experimental Model of Multiple Sclerosis; a Histopathological and Immunohistochemical Study.

Background: Animal model of multiple sclerosis is a demyelinating and inflammatory disorder of central nervous system and eye. Histological evaluation in eyes in experimental autoimmune encephalomyelitis (EAE) models demonstrated evidence of retinal atrophy and inflammation in late stage of disease. Deciphering the relationships between the retinal atrophy and proliferation on retinal layers may help us in understanding the factors that drive atrophy and proliferation in multiple sclerosis. Aims: The aim of the present study was to determine alterations in thickness of retina and its sub-layers in MS induced mice model in comparison with control group. Study Design: Experimental study. Methodology: EAE was induced in female C57BL/6 mice using Hooke kits. Animals were scored for clinical signs of the disease according to 10-point EAE scoring system. At 35th day after immunization, mice (n=15/group) were deeply anesthetized and eyes were removed. Morphometric study of proliferation in retinal sub-layers were assessed by Hematoxilen and Eosin staining and expression of Ki67. The proliferating marker was performed by Ki67analyzing kit. All measurement obtained by Motic image analyzer software 2 and analyzed spss 18, respectively. Results: Here we reported that retinal thickness in MS group including total retinal layer, especially photoreceptor layer, ganglion cell layer and neural layer reduced in comparison to control group (p< 0.001). In Ms Group proliferation rate is also decreased in comparison to control group (0.05). Conclusion: Our results show that ki67 expression, as an indication of proliferation, decreased in retinal layers in MS group. Furthermore, our data revealed that retinal thickness also decreased in MS group.

Evaluation of the melatonin and oxidative stress markers level in serum of fertile and infertile women

Infertility is defined as the inability to achieve the pregnancy within a year of unprotected intercourse. Infertility is a complex issue and different factors such as stress oxidative can be involved in this problem. So, any attempt to neutralize oxidative stress would be helpful in the treatment of infertility. Melatonin is a known scavenger of free radicals. The aim of our study was to evaluate the level of melatonin and its correlation with oxidative biomarkers in fertile and infertile women. The participants including fertile and infertile women were divided into two groups of 30 people. Blood sampling was performed and sera were collected. The level of Malondialdehyde [MDA], total antioxidant capacity [TAG] and melatonin were detected. Data were analyzed using T-test and their correlation was assessed using Spearman test. Serum melatonin from fertile women was higher than infertile women but the difference was not significant [p= 0.46]. MDA level in fertile women was significantly lower than infertile women [p<0.001] and the level of TAG in fertile women was significantly higher than infertile women [p<0.001]. Spearman test revealed a significant and direct correlation between melatonin and TAG in fertile and infertile women and a significant but reverse correlation between melatonin and MDA in infertile and fertile women. Differences in the level of oxidative stress biomarkers in fertile and infertile individuals have been reported. This study revealed a significant correlation between melatonin and oxidative stress biomarkers, concluding that melatonin level could be involved in infertility

MS 14 down-regulates lipocalin2 expression in spinal cord tissue in an animal model of multiple sclerosis in female C57BL/6

Experimental autoimmune encephalomyelitis [EAE] is an animal model of multiple sclerosis, which is a demyelinating and an inflammatory disease of central nervous system. Recent studies have established that some molecules such as Lipocaline2 [LCN2], which expresses during inflammatory conditions, play an important role in EAE pathogenesis and might involve in its treatment process. Recently, it has been proved that MS 14, an herbal-marine drug, has anti-inflammatory properties through reduction of TNF-a and IL-lp. Thus, the present study investigated the effects of MS 14 on the course of EAE and its relation to LCN2 expression in both protein and gene levels. EAE was induced in female C57BL/6 mice using Hooke kits. Animals were scored for clinical signs of the disease according to a 10-point EAE scoring system. On 21[st] and 35[th] days after immunization, mice [n = 4/group] were deeply anesthetized, and the spinal cords were removed. Inflammatory cell infiltration and LCN2 expression in spinal cord were assessed by hematoxylin and eosin staining, immuno-histochemistry, and real-time PCR methods. MS 14 significantly ameliorated EAE symptoms and decreased lymphocyte infiltration into the spinal cord [P<0.05]. Our data also revealed that LCN2 expression was significantly down-regulated in acute and chronic phases of EAE both at protein and gene levels after MS 14 treatment [P<0.05]. The results demonstrated that MS 14 regulatory effect on EAE is accompanied by LCN2 down-regulation after treatment with the herb; however, more studies are required for clarifying the other involved mechanisms

Study of ultrastructure and apoptosis in the endometrium of women with or without endometriosis

More than 40% of infertilities are due to endometriosis. Ultrustructural and histochemical study of endometrium will help to clarify the etiology of endometriosis. The aim of the present study was to investigate the ultrastructure and occurrence of apoptosis in endometrial cells of women with or without endometriosis. In the present case-control study, endometrial specimens from 12 women without endometriosis [as control] and 12 women with endometriosis [as case] were examined. Specimens for control group were obtained from the patients that were referred to gynecology hospital for hysterectomy due to various reasons. In case group the endometriosis was diagnosed according to laparoscopy and endometrial samples were taken using pippel biopsy. The specimens from both case and control groups were processed for Transmission Electron Microscopy [TEM], TUNEL reaction technique and morphometric studies. The results show that endometrial epithelium lost its continuity in women with endometriosis and endometrial cells have euchromatic nucleus in comparison to those from non-endometriosis. There were several apoptotic cells in the luminal and glandular endometrial epithelium and stroma from endometrium of control group. However, apoptotic cells were rarely seen in the endometrium from women with endometriosis. The difference in number of apoptotic cells between two groups statically was significant [p<0.001]. Regarding the ultrastructural characteristics of endometrial epithelial cells and comparison of apoptotic occurrence in control and case groups it is concluded that endometrial cells in endometriosis group have higher potential to survive and possibly implant.

Evaluation of chemoprevention effect of systemic celecoxib on induction of tongue neoplasms in rat

Cyclo-oxygenase-2 [COX-2] specific inhibitors were examined for predication or treatment of different tumors and it is indicated that COX-2 specific inhibitors play an important regulatory role in apoptosis of tumoral tissues. Therefore, the present study was designed in order to examine the preventive effects of a COX-2 specific inhibitor called. celecoxib on 4-nitroquinoline 1-oxide [4NQO]-induced squamous cell carcinoma on rat. In this experimental study, 30 Sprague Dawley rats [with the age of 3- 3.5 months] were selected and divided into three groups. In order to induce lingual carcinoma, 4NQO powder was prepared 3 times a week for each cage. In this study, celecoxib power was mixed with a basic food [basal diet] in order to examine the systematic effect. Tongue samples were sent to laboratory for immunohistochemical [IHC] staining and histological examination. Based on morphological criteria and the ratio of apoptosis to cell proliferation, the prevalence of tongue precancerous lesions was reduced significantly by celecoxib. Celecoxib systematic has inhibitory effects on the 4-nitroquinoline 1-oxide [4NQO]-induced squamous cell carcinoma of tongue. The effect of celecoxib is probably via suppression of cell proliferation and induction of apoptosis

Ultrastructural and morphometrical study of preimplantation endometrium in superovulated mice treated with progesterone or sildenafil

Endometrial development has an important role in blastocyst adhesion and implantation. During IVF cycles, endometrial development is enhanced by progesterone. The aim of this study was to compare ultrastructural and morphometrical characteristics of mice uterine endometrium in natural cycle with those in superovulated cycles received progesterone or Sildenafil. In This study, 60 female bulb/c mice were divided into 4 groups: a control and 3 experimental; gonadotropin, gonadotropin+ Sildenafil and gonadotropin+ progesterone. In experimental groups the mice superovulated mated. In the gonadotropin+ progesterone and gonadotropin+ Viagra groups, the mice respectively received 1mg progesterone and 3 mg Sildenafil citrate. Their uterine specimens were prepared for morphometrical and ultrastructural study. Height of the epithelial cells was measured, using motic software. Statistical analysis was performed using ANOVA. Microscopy revealed that in control group the cells had numerous apical microvilli and the height of the cells was 20.52 +/- 2.43 microm. In gonadotropin+ progesterone group, the granules were found in basal and apical portions and cellular height were 17.91 +/- 2.78 microm which were significantly shorter than in the control and gonadotropin groups [p<0.001]. In this group, the apical membrane also contained pinopodes. In gonadotropin +Sildenafil group, the granules were found in both apical and basal portions and the height of the cells were 17.60 +/- 2.49 microm which were significantly shorter than in the control and gonadotropin groups [p<0.001]. In this group, pinopodes appeared slightly extensive than the other groups. It is concluded that superovulatory drugs in mice stimulate endometrial maturation but injection of Sildenafil is nearly more positive

[Effects of knotweet or polygonum aviculare herbal extract on proliferation of HeLa cell line]

Cervical cancer is the most common cancer in women living in developing countries. Recently, for treatment of diseases such as cancer, herbal medicine is used as a supplementary. The aim of this study was assessment of anticancerous effects of polygonum aviculare herbal extract on Hela cervical cancer cell line. HeLa cells were cultured in RPMI - 1640 with 10% Fetal Bovine Serum in 5% Co2 and at 37 [degree sign] C in different concentrations [0, 0.005, 0.05, 0.01, 0.025, 0.075, 0.1, 0.125, 0.15, 0.175, 0.2, 0.25, 0.3, 0.35, 0.5, 5 mg/ml] of polygonum aviculare. For assessment of viability of cells, trypan blue staining was performed. MTT assay was used for proliferation detection. Our results showed that in 0.15, 0.20 and 0.35 mg/ml proliferation of HeLa cells decreased according to MTT assay. It was proved that polygonum aviculare had antioxidant component and could be a scavenger of free radical. Because of high production of free radicals in diseases such as cancer, the use of the herbal medicine with high amount of antioxidant could be a supplementary treatment in cancer and other diseases

Effect of polyunsaturated fatty acids on secretory phospholipase A2 type IIa in ectopic endometrial cells

Endometriosis is a common chronic inflammation which leads to infertility and chronic pelvic pain in affected women. Secretory phospholipase A2 type IIa [sPLA2IIa] is an acute phase reactant that is markedly increased in inflammatory disorders. To assess the effects of omega-3 and omega-6 polyunsaturated fatty acids [PUFAs] administration in endometrial cells culture on sPLA2IIa level and cell survival comparing homolog ectopic versus eutopic endometrial cells from endometriosis patients. In this experimental study, ectopic and eutopic endometrial tissue samples obtained from 15 endometriosis patients were immediately frozen. After thawing and tissue digestion, mixed stromal and endometrial gland cells were cultured for 8 days in three different culture media; balanced omega-3/omega-6, high omega-3 and high omega-6 PUFAs ratio. Cell survival was measured using 2, 3-bis [2-methoxy-4-nitro-5-sulfophenyl]-5-[phenylamino] carbonyl-2H- tetrazolium hydroxide [XTT] method and sPLA2IIa level assessed with ELISA technique. The sPLA2IIa level was significantly higher in the ectopic endometrial cell culture compared to the eutopic group for each of the three matched treatments [balanced, high omega-3 and high omega-6]. Also the sPLA2IIa level in the ectopic endometrial cell group was remarkably increased by each of the three PUFAs treatments compared to control condition [p<0.05, p<0.01, p<0.05 respectively]. Cell survival in the eutopic group was significantly decreased by high omega-6 culturing compared to control medium [p<0.05]. The increase in sPLA2IIa level in ectopic endometrial cells by fatty acid treatments [especially high omega-3], strengthens the hypothesis that PUFAs stimulate secretion of cytokines leading to increased sPLA2IIa level

Efficacy of hepatitis B vaccine in those who lost hepatitis B surface antigen during follow-up

The level of HBsAg in some chronic hepatitis B virus [HBV]-infected individuals may decline over time so that it is not detectable in serum. To assess the efficacy of HBV vaccine in those who lost their HBsAg without seroconverssion to anti-HBs antibody. From April 1993 to December 2008, of 1603 chronic HBV-infected individuals, 34 [22 men and 12 women] became HBsAg-negative in follow-up visits, with no detectable anti-HBs antibody and HBV DNA in their sera. They received HBV vaccination at 0, 1 and 6 months [case group]. Fifty-two subjects [30 men and 22 women] who were negative for HBsAg, anti-HBs and anti-HBc antibody, received HBV vaccination according to the said schedule [control group]. Anti-HBs antibody was assessed one month after the last dose of vaccination in the both groups. The mean +/- SD age of the case and control groups was 38 +/- 12.7 and 33.4 +/- 8.6 years, respectively [p=0.07]. The sex distribution between these two groups were similar [p=0.652]. The mean +/- SD years of follow-up for the case group was 7.6 +/- 4.5 years. Anti-HBs antibody level >/= 10 IU/L was found in 8 [24%] subjects in the case group and in 45 [87%] in the control group [p<0.001]. The mean +/- SD anti-HBs antibody level in the case group was 68 +/- 32.66 and in the control group 344.6 +/- 38.9 IU/L [p<0.001]. We found that nearly 24% of chronic HBsAg-positive subjects who lost their HBsAg responded to HBV and the remaining cases need to be followed for occult HBV infection

High frequency electromagnetic field induces lipocalin 2 expression in mouse liver

Neutrophil gelatinase-associated lipocalin [NGAL/Lcn2], comprise a group of small extracellular proteins with a common beta-sheet-dominated 3-dimensional structure. In the past, it was assumed that the predominant role of lipocalin was acting as transport proteins. Recently it has been found that oxidative stress induces Lcn2 expression. It has been also proved that electromagnetic field [EMF] produces reactive oxygen species [ROS] in different tissues. Expression of Lcn2 following exposure to electromagnetic field has been investigated in this study. Balb/c mice [8 weeks old] were exposed to 3 mT, 50 HZ EMF for 2 months, 4 hr/day. Afterwards, the mice were sacrificed by cervical dislocation and livers were removed. The liver specimens were stained with Haematoxylin-Eosin [H and E] and analyzed under an optical microscope. Total RNA was extracted from liver and reverse transcription was performed by SuperScript III reverse transcriptase with 1 micro g of total RNA. Assessment of Lcn2 expression was performed by semiquantitative and real time-PCR. The light microscopic studies revealed that the number of lymphocyte cells was increased compared to control and dilation of sinosoids was observed in the liver. Lcn2 was up-regulated in the mice exposed to EMF both in mRNA and protein levels. To the extent of our knowledge, this is the first report dealing with up-regulation of Lcn2 in liver after exposure to EMF. The up-regulation might be a compensatory response that involves cell defense pathways and protective effects against ROS. However, further and complementary studies are required in this regards