new indole derivative decreased SALL4 gene expression in acute promyelocytic leukemia cell line [NB4]

Background: Acute myeloblastic leukemia [AML] is a clonal disorder due to bone marrow failure and uncontrolled proliferation of myeloid lineage. Acute promyelocytic leukemia [APL] is a subtype of AML. Heterocyclic compounds, such as indole, are considered as attractive candidates for cancer therapy, due to their abundance in nature and known biological activity. Sal-like protein [SALL4] is a zinc finger transcription factor involving in the multi-potency of stem cells, in the NB4 cell line. This study was aimed to evaluate the effects of basal indole and its new derivative, 2-[1-[[2, 4-Aril]imino]-2,2,2-trifluoroethyl] phenyl-1H Indole-3- carbaldehyde [TFPHC], on the expression of SALL4

Methods: Cells were cultured and treated with different concentrations [75, 150, and 300 micro g/mL] of the new indole derivative and DMSO, as a vehicle control, for 24 and 48 hours. Cell proliferation was evaluated by using Trypan blue exclusion and MTT assays. The percentage of apoptotic cells was determined by flowcytometry analysis using the Annexin V/PI apoptosis detection kit; mRNA expression of SALL4 was studied using absolute quantitative RT-PCR

Results: Our findings demonstrated the effects of new indole derivatives on SALL4 mRNA expression. Expression of SALL4 mRNA was significantly decreased at 75, 150, and 300 micro g/mL concentrations

Conclusion: SALL4 plays a role in the survival of APL cells. SALL4 expression could be suppressed by the novel indole derivative. Additionally, SALL4 gene suppression can serve as a target in APL therapy

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

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

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

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

Exercise preconditioning decreases deleterious effects of transient cerebral ischemia in rat's

Background: Transient global cerebral ischemia causes extensive neuronal damage in the brain and leads to a deficit in learning and memory. We designed the present study to investigate the effect of exercise preconditioning on learning and spatial memory following transient cerebral ischemia in rat

Materials and methods: In this experimental study, 50 male Wistar rats weighing 250-300g were randomly allocated to different groups. Exercise was done by treadmill and, for inducing cerebral ischemia, both common carotid arteries were occluded for 10 minutes. Memory was evaluated using a step-through passive avoidance task. Sensory motor deficits were assessed by adhesive removal test. For evaluating slip ratio, we used ledged beam walking test

Results: One week after transient cerebral ischemia, response latency decreased in passive avoidance test. Also touch time, remove time, and slip ratio were increased in these animals. Exercise preconditioning improved the measured indices in ischemic rats

Conclusion: Exercise preconditioning improved deficits in learning and memory, as well as sensory-motor function, following transient cerebral ischemia

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

Non-association of IL-12 +1188 and IFN-gamma +874 polymorphisms with cytokines serum level in occult HBV infected patients

Occult hepatitis B infection [OBI] is identified as a form of hepatitis in which despite the absence of detectable HBsAg, HBV-DNA is observed in peripheral blood of patients. The main aim of this study has been to investigate the association between polymorphisms in +874 of IFN-gamma and +1188 of IL-12 with their serum level in patients suffering from OBI. In this experimental study, plasma samples of 3700 blood donors were tested for the presence of hepatitis B surface antigen [HBsAg] and anti-HBc by ELISA. The HBsAg[-]/anti-HBc[+] samples were selected and screened for HBV-DNA by PCR. HBV-DNA positive samples were assigned as OBI cases and ARMS-PCR techniques were performed to examine the two known polymorphisms within IL-12 and IFN-gamma. In addition, the serum levels of IL-12 and IFN-gamma were also determined by ELISA. Results of this study demonstrated that, 352 [9.5%] out of 3700 blood samples were HBsAg[-]/anti-HBc[+]and HBV-DNA was detected in 57/352 [16.1%] of HBsAg[-]/anti-HBc[+] samples. Our results showed that groups showed significant difference in CC allele of +1188 region of IL-12 and no difference was observed in the other evaluated genes. Our results also showed that the alleles of +1188 region of IL-12 and alleles of +874 of IFN-gamma were also not associated with serum level of cytokines. According to the results of this study, it may be concluded that the polymorphisms in +1188 region of IL-12 and +874 region of IFN-gamma would not affect the expression of both cytokines at serum level in OBI patients

Impact of opium on the serum levels of TGF-beta in diabetic, addicted and addicted-diabetic rats

Several cells of immune system such as regulatory T cells and macrophages secrete transforming growth factor-beta [TGF-beta] in response to different stimuli. This cytokine has inhibitory effect on immune system and diminished production of this cytokine is associated with autoimmune disorders. The aim of this study was to evaluate the influence of opium addiction on serum level of TGF-beta in male and female diabetic and non-diabetic Wistar rats. This experimental study was performed on normal, opium addicted, diabetic and addicted-diabetic male and female rats. Serum level of TGF-beta was measured by ELISA. The results of our study indicated that the mean serum level of TGF-beta in female addicted rats was significantly increased compared to control group [p<0.004]. Conversely, in male addicted rats the mean serum level of TGF-beta was lower compared with control [p<0.065]. Our results suggest that opium and its derivatives have differential inductive effects on the cytokine expression in male and female rats

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

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

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

[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

Expression of stromal derived factor alpha [SDF-1a] by primary hepatocytes following isolation and heat shock stimulation

Stromal derived factor-alpha [SDF-1-alpha] is a CXC chemokine which has been demonstrated as a recruitment factor for leukocytes to the site of inflammation, infection, injury and following stress. This chemokine has been shown to be expressed by liver cells and in liver diseases. Hence, the aim of this study was to examine the expression of SDF-1 by hepatocytes in responses to the stress imposed during isolation by collagenase perfusion and under heat shock stimulation. In this study hepatocytes [2-5 x 10[6]] were isolated from male Sprague Dawley rat liver and cultured in plates that were pre-coated with collagen Type-I matrix. The western and northern blotting analysis were employed to detect SDF-1 at protein and mRNA levels in isolated and cultured hepatocytes in response to isolation and heat shock stresses. The SDF-1 is expressed by isolated rat hepatocytes immediately after isolation and early culture and decreased with time. SDF-1 protein was highly expressed in freshly isolated cells and decreased by time [27h] [P < 0.05]. mRNA was also expressed in freshly isolated cells [0h] but decreased after 24h of culture [P < 0.01]. This results also demonstrated that expression of SDF-1 by hepatocytes was increased in response to heat shock at different time points comparing with control [P < 0.01]. These results demonstrated that the isolation and heat shock stresses induced the expression of SDF-1 in hepatocytes in a time-dependent manner. Accordingly, it seems that hepatocytes mimic the experiences that liver experience after injury in vivo and therefore, produce stress related agents like chemokines to overcome such a injurious condition

Expression of IP-10 chemokine is regulated by pro-inflammatory cytokines in cultured hepatocytes

Chemokines are classified in four distinct groups as CXC, CC, CX3C and C, depending on the presence or absence of a motif called ELR [Arg-Leu-Glu] before the first cysteine residue in their structure. CXC chemokines are also subdivided into ELR+and ELR-. Increasing evidence has indicated the existence of a chemokine network in the liver which is involved in both physiological responses and, under certain circumstances, pathological and repair processes following hepatic injury. The CXC chemokines play a major role in both these processes, and much attention has been focused on their therapeutic applications to liver disease. The aim of this study was to examine the response of cultured hepatocytes to exogenous inflammatory cytokines [TNF-alpha and IFN-gamma] regarding expression of IP-10 and growth regulatory oncogen [Gro] chemokines. In this study we employed western and northern analysis to measure chemokines at the level of protein and mRNA by hepatocytes in response to pro-inflammatory cytokines. We found that, the pro-inflammatory cytokines, TNF-alpha and IFN-gamma, selectively stimulated expression of IP-10 but were without effect on Gro. This confirms a potential direct involvement of these cytokines in chemokine production by hepatocytes. Thus, IFN-gamma and TNF-alpha may play a role in hepatic injury and inflammation and produce some of their biological effects by localized induction of chemokines by hepatocytes. Given the similarity to an acute phase response, we were able to show that IFN-gamma and TNF-alpha mimicked the effects of cell isolation and culture on induction of IP-10 expression. Further, evidence for linkages between IFN-gamma and TNF-alpha and liver injuries is seen in hepatitis C and hepatitis B in which increased levels of TNF-alpha and its soluble receptor were reported

Expression of cxc chemokines Gro/Kc and SDF-1 alpha in rat H4 h epatomacells inresponse to different stimuli

It is now well established that several environmental stress factors cause activation of p38 MAP kinase and JNK in various cell types to produce chemokines. To investigate the expression of CXC chemokines Gro/KC and SDF- 1alpha in rat's H4 hepatoma cells in response to heat shock, hyperosmolarity and oxidative stress. Hepatoma cells were maintained in MEM medium. Cells were subjected to different stresses [[H[2]O[2] 0.15% [w/v], manitol and NaCl [160 mM] and heat shock [42 °C for 20 minutes]]. Cells were harvested and RNA was extracted, purified and the CXC chemokine Gro/KC and SDF-1alpha expression was analysed by RT-PCR. cDNA was separated by gel electrophoresis on a 1% [w/v] agarose gel and visualized under a UV transilluminator. There was detectable but low expression of both SDF-1 alpha and Gro/KC in H4 hepatoma cells. Heat shock failed to induce expression of SDF-1alpha and Gro/KC in H4 hepatoma cells of rat. Hyperosmolarity also did not stimulate SDF-1alpha and Gro/KC expression. In this study we have also shown that oxidative stress did not induce expression of SDF-1alpha and Gro/KC. Overall, although detection is possible but regulatory responses were not observed in H4 hepatoma cells. Several known injurious conditions cause recruitment of macrophages, neutrophils and other immune cells to the liver. Immune cells are recruited to the hepatic vasculature following local liver injury and subsequent chemokine production. Our results demonstrated that failure to produce chemokines by hepatoma cells may be a way to escape from mechanism of immune surveillance