Can CRP/melatonin ratio measurement be used as a predictor of multiple sclerosis?

@#Background & Objective: This study aimed to find a biomarker to predict the development of multiple sclerosis (MS). Serum levels of vitamin D3, C-reactive protein (CRP) and melatonin and their ratio were evaluated to find the valuable cut-off point. Methods: Serum levels of vitamin D3, CRP and melatonin were evaluated using commercial ELISA kit in newly diagnosed MS patients and compared with healthy controls. Results: Serum CRP level significantly increased and serum melatonin level significantly decreased in MS patients in comparison to controls. Sensitivity, specificity, positive predictive value, negative predictive value and accuracy for the cut-off point of CRP/melatonin ratio ≥ 78.29087 were 80%. Conclusion: CRP/melatonin ratio ≥ 78.29087 may be used for prediction of MS in an at risk population

Altered expression of high molecular weight heat shock proteins after OCT4B1 suppression in human tumor cell lines

Objective: OCT4B1, a novel variant of OCT4, is expressed in cancer cell lines and tis-sues. Based on our previous reports, OCT4B1 appears to have a crucial role in regulating apoptosis as well as stress response [heat shock proteins [HSPs]] pathways. The aim of the present study was to determine the effects of OCT4B1 silencing on the expression of high molecular weight HSPs in three different human tumor cell lines

Materials and Methods: In this experimental study, OCT4B1 expression was suppressed in AGS [gastric adenocarcinoma], 5637 [bladder tumor] and U-87MG [brain tumor] cell lines using RNAi strategy. Real-time polymerase chain reaction [PCR] array was employed for expression level analysis and the fold changes were calculated using RT2 Profiler PCR array data analysis software version 3.5

Results: Our data revealed up-regulation of HSPD1 [from HSP60 family] as well as HSPA14, HSPA1L, HSPA4, HSPA5 and HSPA8 [from HSP70 family] following OCT4B1 knock-down in all three cell lines. In contrast, the expression of HSP90AA1 and HSP90AB1 [from HSP90 family] as well as HSPA1B and HSPA6 [from HSP70 family] was down-regulated under similar conditions. Other stress-related genes showed varying expression pattern in the examined tumor cell lines

Conclusion: Our data suggest a direct or indirect correlation between the expression of OCT4B1 and HSP90 gene family. However, OCT4B1 expression was not strongly correlated with the expression of HSP70 and HSP60 gene families

Vitex Agnus Castus extract improves learning and memory and increases the transcription of estrogen receptor alpha in hippocampus of ovariectomized rats

Introduction: Lower level of estrogen hormone is considered as an important factor for loss of learning and memory in postmenopausal women. Although estrogen replacement therapy is used for compensation, but long-term usage of estrogen is associated with a higher risk of hormone-dependent cancers. Phytoestrogens, due to fewer side effects, have been proposed to prevent menopause-related cognitive decline

Methods: 24 female Wistar rats weighing 180-220 g were used in this study. The animals were ovariectomized and randomly divided into four groups including, control and two groups which received 8 and 80 mg/kg Vitex agnus castus [VAC] ethanolic extract orally. The last groups were treated with 40 micro g/kg of estradiol valerat. Step-through passive avoidance [STPA] test was used for the evaluation of learning and memory. The hippocampal estrogen receptor alpha [ERalpha] expression was measured using Real-Time PCR

Results: The results demonstrated that VAC extract or estradiol had better performance on step-through passive avoidance test than control group [all P<0.05]. Moreover, administration of either estradiol or VAC extract increased the hippocampal mRNA level of ER alpha and prevented the decrease in uterine weight of ovariectomized rats

Discussion: Based on our data, VAC extract improves learning and memory in ovariectomized rats. The positive effect of VAC extract on learning and memory is possibly associated with an increase in ER alpha gene expression in the hippocampal formation

Molecular and serological detection of acute and latent toxoplasmosis using real-time PCR and ELISA techniques in blood donors of Rafsanjan city, Iran, 2013

The differentiation between acute and latent forms of the Toxoplasma gondii [T. gondii] infection is still considered as a complicated issue. This study was aimed to elucidate the status of infection in the blood donors and the probable importance of blood transfusion in the transmission of the infection through detecting both immunological and genetic markers of acute and latent infection. Totally 235 blood samples from blood donors were collected. The levels of anti-T. gondii IgG and IgM antibodies were examined by specific ELISA kits. cDNA were synthesized from total extracted mRNA molecules from the serum samples and SAG1 gene, specific for tachyzoite form, were amplified using Real-Time PCR technique. Demographic information of study subjects including their gender, age, job, and habitat were recorded. Out of 235 serum samples, 80 [34.04%] and 4 [1.71%] were positive regarding anti-T. gondii IgG and IgM antibodies, respectively. Real-Time PCR results showed that 14 out of 200 [6.97%] of blood donor had mRNA molecules of SAG1 gene. The positive results of Real-Time PCR of SAG1 in female gender and housekeepers were significantly higher than those of male gender and other job categories. The prevalence of chronic and acute infection is high in Iranian blood donors. Additionally, evaluation of antibodies could not be reliable, because several donors negative for anti-T. gondii IgM antibodies had detectable SAG1 mRNA molecules. Hence, it seems that molecular diagnostic tests are essential to detect acute infections

Morphine reduces expression of TRPV1 receptors in the amygdala but not in the hippocampus of male rats

Background: Chronic use of opioids usually results in physical dependence. The underlying mechanisms for this dependence are still being evaluated. Transient receptor potential vanilloid type 1 [TRPV1] are important receptors of pain perception. Their role during opioid dependence has not been studied well. The aim of this study was to evaluate the effect of morphine-dependence on the expression of TRPV1 receptors in the amygdala and CA1 region of the hippocampus

Methods: This study used four groups of rats. Two groups of rats [morphine and morphine+naloxone] received morphine based on the following protocol: 10 mg/kg [twice daily, 3 days] followed by 20, 30, 40 and 50 mg/kg [twice daily], respectively, for 4 consecutive days. Another group received vehicle [1 ml/ kg] instead of morphine given using the same schedule. The morphine+naloxone group of rats additionally received naloxone [5 mg/kg] at the end of the protocol. The control group rats received no injections or intervention. The amygdala and CA1 regions of the morphine, saline-treated and intact animals were isolated and prepared for real-time PCR analysis

Results: Administration of naloxone induced withdrawal signs in morphine-treated animals. The results showed a significant decrease in TRPV1 gene expression in the amygdala [P<0.05] but not the CA1 region of morphine dependent rats

Conclusion: TRPV1 receptors may be involved in morphineinduced dependence

Prevalence of hepatitis B co-infection among HIV positive patients: narrative review article

Hepatitis B virus [HBV] is the most prevalent viral infection and is among the leading causes of human liver diseases. Nearly 360 millions of people are world widely infected with prolonged forms of hepatitis B including active and inactive chronic forms. Chronic hepatitis B [CHB] is associated with cirrhosis and hepatocellular carcinoma [HCC] in patients suffering from congenital and/or acquired immunodeficiency and also following immunosuppressive therapy. The target cell of human acquired immunodeficiency virus [HIV] is CD4 positive T cells. These cells play central role[s] in both cellular and humoral immunity so that the HIV attack of CD4 positive T cells causes suppression of both cell-mediated and humoral immune responses. One of the frequent complications in HIV positive patients is HBV co-infection and as a result, the co-transmission of these viral diseases is common. Due to the paramount importance of the co-infection of HBV and HIV, it is noteworthy to investigate the prevalence of hepatitis B in these patients for planning of an effective therapeutic strategy. Based on these considerations, the main aim of this review article was to collect and analyze the recent and relevant studies regarding the prevalence rate of hepatitis B co-infection among HIV positive patients world widely

Interleukin-10 serum levels after vaccination with in vivo prepared Toxoplasma gondii excreted/secreted antigens

Toxoplasma gondii is a worldwide prevalent zoonotic parasite which causes toxoplasmosis. An appropriate vaccine for animals could interrupt the circle between animals and humans. Our previous study showed that excreted/secreted antigens [E/ SA], derived from the peritoneum of mice infected with T. gondii tachyzoites could be considered as a good candidate for animal vaccination. Interleukin-10 [IL-10] inhibits proliferation of B and T lymphocytes and induces homeostasis in immune system responses. However, since IL-10 has also been shown to suppress the killing of T. gondii by human macrophages, the aim of this study was to evaluate IL-10 serum levels after vaccination with T. gondii E/SA prepared in vivo. T. gondii tachyzoites were inoculated in the peritoneum of mice and harvested E/SA were used as a vaccine, with and without adjuvant, in T. gondii infected and un-infected mice. IL-10 serum levels were evaluated using the ELISA technique. The data showed that although serum levels of IL-10 were not changed at the early phases, they were elevated at the end phases of vaccination with T. gondii E/SA. Based on these and our previous results, it can be concluded that in vivo prepared T. gondii E/SA could be considered as a good candidate for animal vaccination

Inflammation and depression: Mechanisms and involved molecules

Depression is a main complication in the developed and in developing countries. Studies showed that depression not only is a emotional disease but also affect the body. Immune system disordering is an important complication following depression. Studies showed that depression can induce immunological chronic inflammation, hence, lead to accelerate depression. Several molecules including JAK/STAT pathways and Toll like receptors, AIMII and NLRs play important roles in inducing chronic inflammation. It seems that the mentioned molecules can induce inflammation via cytokine production in depressed patients. Therefore, evaluation of cytokine patterns and cytokines producing pathways in depressed patients can help scientists to find the pathogenesis of depression. Thus, the aim of this review article was to collect recent information regarding cytokine patterns as well as the related molecules in inflammation in depression

Differential expression of CXCL1, CXCL9 CXCL10 and CXCL12 chemokines in alopecia areata

Alopecia Areata [AA] is a non-scarring, autoimmune disorder which causes hair loss. Inflammatory reactions are involved in hair loss of the scalp and/or body. The involvement of chemokine receptors in the pathogenesis of AA is well defined among which, CXCL1 acts on neutrophils and CXCL9, CXCL10 and CXCL12 and serve as T lymphocytes recruiters. To study the serum levels of ELR+ and ELR- CXCL1, CXCL9, CXCL10 and CXCL12 in the patients suffering from AA and healthy controls. The study population of consisted of 30 patients suffering from AA and 30 healthy controls. Serum concentrations of CXCL1, CXCL9, CXCL10 and CXCL12 were measured using enzymelinked immunosorbent assay [ELISA]. Current results showed that AA patients had significantly elevated serum levels of CXCL9 and CXCL10 in comparison to controls [p<0.001]. These results also demonstrated that serum levels of CXCL1 and CXCL12 were significantly decreased in AA patients compared to control [p<0.001]. CXCL9 and CXCL10 are elevated in the AA patients and may be involved in the recruitment of T lymphocytes to the inflamed tissues. Moreover, due to the significant role played by these chemokines in angiogenesis/ angiostatis phenomenon they could be considered as useful biomarkers in AA diagnosis and therapy

MiR-143 induces expression of AIM2 and ASC in jurkat cell line

Absent in Melanoma 2 [AIM2] is an intracellular microbial dsDNA sensor which plays an important role in production of proinflammatory cytokines through Apoptosis associated Speck-like protein containing a Caspase activation and recruitment domain [ASC] and Caspase-1. Micro-RNAs [miRNAs] play important roles in regulation of immune related genes. However, there is little information regarding the effects of miRNAs on the AIM2 and ASC expression. To determine the mRNA levels of AIM2 and ASC in Jurkat cell line following introducing miRNA-143 [MiR-143]. MiR-143, a scrambled sequence and PBS were introduced separately, to the Jurkat cell lines and the mRNA levels of AIM2 and ASC were examined in parallel with beta-actin and GAPDH [as housekeeping genes] using Real-Time PCR technique. The mRNA levels of AIM2 and ASC were significantly increased in the MiR-143 transfected Jurkat cells when compared to the scrambled sequence or PBS treated cells. MiR-143 can lead to increased expression of AIM2 and ASC mRNAs. Considering the significance of AIM2 and ASC in DNA sensing and inflammosome formation, it can be considered as a therapeutic agent for the treatment of chronic infectious diseases, especially viral infections

Association of interleukin-4 polymorphisms with multiple sclerosis in southeastern Iranian patients

Immune system-related factors are important in the pathogenesis of multiple sclerosis [MS]. Interleukin 4 [IL-4] as a helper T cell [2TH] cytokine is involved in the regulation of immune responses. Hence, this study was designed to explore the association between MS and polymorphisms in the -590 region of IL-4. A descriptive study at Rafsanjan University of Medical Sciences, Rafsnajan from September 2009 to August 2010. Blood samples were collected from 100 MS patients and 150 healthy controls on EDTA precoated tubes. DNA was extracted and analyzed for IL-4 polymorphisms using restricted fragment length polymorphism in patients and controls. Demographic data were also collected by a questionnaire that was designed specifically for this study. We observed a significant difference in the C/C, T/C, and T/T genotypes of the -590 region of IL-4 between patients with MS and healthy controls [P<.001]. We conclude that functional polymorphisms of IL-4 possibly play a crucial role in the pathogenesis of MS

Polymorphisms within exon 9, but not intron 8, of the vitamin d receptor gene are associated with Asthma

Deregulation of the immune system through allied factors and cytokine responses are thought to be important contributors to the pathogenesis of asthma. Vitamin D3 and its nuclear receptor appear to be factors that maybe involved in regulating i mmune responses during the progression of asthma. The aim of this study was to investigate the association between polymorphisms in intron 8 and exon 9 of the vitamin D receptor [VDR] and this disease. This study was performed on 100 asthmatic patients and 100 healthy controls. PCR-RFLP was performed to examine polymorphisms in intron 8 and exon 9 of VDR gene. Our results showed a statistically significant difference in the Taq-1 evaluated genotypes of exon 9 of the VDR gene when comparing healthy patients to asthmatic patients. Based on our results, it can be concluded that VDR and its functional polymorphisms may play an important role in the pathogenesis of asthma

Common HBV genotype in southeastern Iranian patients

Asymptomatic hepatitis B infection is characterized as a type of hepatitis in which hepatitis B surface antigen is present in the patient's peripheral blood despite the absence of clinical symptoms. Previous studies have shown that a particular genotype may effect clinical manifestations of hepatitis B infection; hence, the aim of the current study was to determine the frequency of hepatitis B virus genotypes among asymptomatic carriers of hepatitis B. In this experimental study, the plasma samples of 100 asymptomatic carriers were collected and tested for HBsAg and anti-HBs using ELISA. The genotype of hepatitis B virus was determined by the GAP-PCR technique. The results of this study showed that all samples were positive for hepatitis B surface antigen and anti-hepatitis B core antigen was present in 60 [60%] cases. Our results also indicated that all patients had the D genotype of hepatitis B virus

in vitro assay to evaluate the immunomodulatory effects of unrestricted somatic stem cells

Unrestricted somatic stem cells [USSC] are cord blood stem cells that have been considered as candidates for the regulation of immune responses. Therefore, potential exists for their use in the suppression of immune response after transplantation surgery. The aim of this study was evaluation of the effect of USSC on mixed lymphocyte reaction [MLR] as a model for graft rejection. USSC and mesanchymal stem cells [MSC] were isolated and cultured from cord blood and bone morrow, respectively. The immunophenotypes of USSC and MSC were evaluated by flow cytometery and USSC and MSC were cocultured with peripheral blood lymphocytes [PBL] in an MLR to evaluate the immunomodulatory effect of these cells as a percentage of the control response. Current study demonstrated that proliferation of lymphocytes in the MLR was decreased after treatment with USSC, in a similar fashion to that seen with MSC. It can be concluded that USSC have similar regulatory effects as MSC on the MLR, which can be used as an indicator for potential organ rejection after transplantation. Therefore, the immunoregulatory effect of these cells could be used in the clinic during organ transplantation and in the management of autoimmunity

Expression of regulated oncogen-alpha by primary hepatocytes following isolation and heat shock stimulation

High levels of regulated oncogen-alpha [GRO-a] expression have been observed in the liver. GRO-a stimulates proliferation of epithelial cells and induction of rolling and extravascular migration of neutrophils and mononuclear cells. Given the above observations, this chemokine was chosen to be analyzed in freshly isolated and cultured hepatocytes. In this study, hepatocytes [2_106 cell/ml] were isolated from male Sprague Dawley rat liver and cultured on plates that were pre-coated with collagen type-I matrix. The western and northern blot analyses were employed to detect GRO-a at the protein and mRNA levels in freshly isolated and cultured hepatocytes in response to isolation and heat shock stresses. GRO-a was shown to be expressed by isolated rat hepatocytes immediately after isolation and early culture and decreased with time. mRNA was also expressed in freshly isolated cells [0 h] and did not decrease after 48h of culture and further time points [P<0.01]. These results also demonstrated that expression of GRO-a by hepatocytes increased in response to heat shock at different time points in comparison with the control [P<0.01]. These results demonstrated that the isolation and heat shock stresses induced the expression of GRO-a in hepatocytes in a time-dependent manner. Thus, it seems that hepatocytes mimic the experiences that the liver encounters after injury in vivo. In such a situation, liver produces stress related agents like chemokines to overcome injurious conditions

Evaluation of relation between IL-4 and IFN-gamma polymorphisms and type 2 diabetes

Although, type 2 diabetes is the most frequent type of diabetes, its main cause is yet to be clarified. Several environmental and genetic parameters are believed to be involved in diabetes. It has also been established that cytokines play key roles in pathogenesis of diabetes. Expression of cytokines is different from person to person and in different societies. Several studies showed that polymorphisms of +874 of interferon-gamma [IFN-gamma] and -590 of interleukin-4 [IL-4] are associated with the regulation of expression of these genes. This study was aimed to find polymorphisms of these regions in type 2 diabetes patients. In this experimental study peripheral blood samples were collected from 160 type 2 diabetic patients and 160 healthy controls. DNA was extracted by salting out method. Polymorphisms of +874 of 1FN-gamma and -590 of IL-4 were analyzed by ARMS-PCR and RFLP-PCR. Our findings indicated that TT genotype of IFN-gamma was increased in type 2 diabetic patients compared to the control but difference was not significant. Our results didn't show any significant difference between IL-4 genotype in diabetic and healthy controls either. Our results suggested that TT genotype of IFN-gamma can be associated with diabetes. This association can be described by the fact that over expression of IFN-gamma shifts immune system to Th 1; therefore, pancreatic cells can be miscarried by immune cells

[Evaluation of expression rate of chemokines receptor CCR5 on peripheral blood CD8[+] T cells of occult hepatitis B infected patients]

Occult hepatitis B infection [OBI] is defined as a form of hepatitis B that despite absence of detectable HBsAg, HBV-DNA is present in patient's peripheral blood. Genetic and immunological differences appear to play important roles in producing OBI. Therefore, this project was aimed to examine the expression of a chemokine receptor [CCR5] on CD8[+] T cells of OBI patients. In this experimental study, 3,700 HBsAg- plasma samples were collected. Samples were tested for anti-HBc antibody and all of HBsAg-/anti-HBc[+] samples were screened for HBV-DNA by PCR. HBV-DNA positive samples were assigned as OBI cases. Also, flow cytometry analysis was performed to examine the expression of CCR5 on CD8[+] T cells of OBI patients. Results of current study showed that 352 [9.5%] cases of samples were positive for anti-HBc. Examination of HBsAg/anti-HBc[+] samples for HBV-DNA by PCR showed that 57 [16.1%] cases had HBV-DNA. Flow cytometric studies indicated lymphocytosis in these patients; however, the number of cells which expressed CD8[+] and CCR5 is decreased significantly in patients, compared to healthy control. In addition to CD8[+] T cells, the expression of CCR5 is also decreased on all immune cells. One of the chemokine receptors which are expressed by CD8[+] T cells is CCR5 and these cells are recruited to infected tissues, including liver by CCR5. Therefore, based on results of this investigation, one may conclude that due to the decreased expression of CCR5, the CD8[+] T cells are unable to respond to the chemokines [CCR5 ligands] and, hence, can not immigrate to the infected liver and incorporate in clearance of hepatitis B virus

[Evaluation of prevalence of delta 32 mutation in CCR5 gene in breast cancer patients in Rafsanjan city]

Chemokines and their receptors are expressed in different types of malignancies. CC chemokines MIP-1alpha [CCL3], MIP-1beta [CCL4] and RANTES [CCL5] is believed to be anti-tumor and also aid to the metastasis in tumor microenvironment. CCR2 and CCR5 are special G-protein receptors for these chemokines. Due to the important role of CCR5 chemokine receptor in tumor biology, this project is designed to examine delta 32 mutation in CCR5 gene regards breast cancer. This experimental study was performed during 2007-8 on delta healthy adults and 36 breast cancer patients by Gap-PCR. The demographic information also was collected by questionner and t-test Chi-square was used for statistical analysis of data. Our results showed that none of breast cancer patients had CCR5-delta 32 mutation while 3 [3%] cases of controls had heterozygotic form of this mutation. Our results showed that there is not any CCR5-delta 32 mutation in patients. Therefore, it appears that this mutation don't play any role in breast cancer

[Evaluation of relation between IL-4 and IFN-gamma polymorphisms and type 2 diabetes]

Although Type 2 diabetes is the most frequent among different types of diabetes, the cause of it is yet to be clarified. Several environmental and genetic factors are said to be involved in diabetes and it has been established that cytokines play key roles in pathogenesis of diabetes. Expression of cytokins is different from person and in different societies. Studies showed that polymorphisms of +874 of IFN-gamma and -590 of IL-4 regions are related to the expression of these genes. In this study, we aimed to find polymorphisms of these regions in Type 2 diabetes patients. In this study peripheral blood samples were collected from 51 type 2 diabetes patients and 50 healthy controls. DNA was isolated by salting out method, using ARMS-PCR, RFLP-PCR polymorphisms of = 874 of IFN-gamma and -590 of IL-4, were analyzed, respectively. Our findings showed that TT genotype of IFN- gamma was increased in type 2 diabetic patients as comford with control group but difference was not significant. Our results also have not shown any significant difference between IL-4 genotype in diabetic and healthy controls. Our results suggested that TT genotype of IFN- gamma can be related to diabetes. This relation can be described by this known Todgment judyunt that over expression of IFN- gamma shifted immune system to Th1, therefore, pancreas cells were miscarried by immune cells

[Occult HBV infection in Rafsanjanese blood donors]

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 of status of occult hepatitis B infection in the Rafsanjanese blood donors. 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 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. 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