Monarch-1 activation in murine macrophage cell line [j774 a.1] infected with Iranian strain of leishmania major

Leishmania major is an intracellular parasite transmitted through the bite of the female phlebotomine sand flies. Leishmania major is able to escape the host immune defense and survive within macrophages. Modulation of the NF-kappaB [Nuclear Factor-Kappa B] activation and suppression of the pro-inflammatory cytokines by L. major are the main evasion mechanisms that remain to be explored. This study aims to examine the expression level of the Monarch-1 in L. major-infected macrophages, as a negative regulator of the NF-kappaB activation. Murine macrophage cell line [J774 A.1] was infected by metacyclic form of Leishmania promastigotes at macrophage/parasite ratio of 1:10. After harvesting infected cells at different times, total RNA was extracted and converted to cDNA. Semi-quantitative RT-PCR was performed for Monarch-1 by specific primers. Hypoxanthine Phospho-Ribosyl Transferase [HPRT] was used as an internal control to adjust the amount of mRNA in each sample. Semiquantitive analysis of Monarch-1 mRNA expression level showed a significant expression increase within 6 to 30 hours after L. major infection of macrophages when compared to the control macrophages. Monarch-1 expression level reveals a significant increase in the early phase of macrophage infection with L. major, which in turn may suppress IL-12 production in Leishmania infected macrophages and deeply influence the relationship between host and parasite.

Apoptosis of human lymphocytes after exposure to hydatid fluid

Modulation of the immune response is an important strategy by which establishment and growth of hydatid cyst in the internal organs of human is warranted. Induction of apoptosis in the lymphocytes might be a considerable component. This study was designed to evaluate apoptotic impact of hydatid fluid [HF] on human lymphocytes. Human lymphocytes were treated with hydatid fluid. After 6 hours of exposure, caspase-3 activity, the central enzyme of apoptosis cascade, was measured by fluorometric assay in the HF-treated lymphocytes and control cells. In addition, the expression of Bax [a pro-apoptotic protein] and Bcl-2 [an anti-apoptotic protein] mRNA was assessed by RT-PCR after 12 hours of exposure. Both the ratio of Bax/Bcl-2 mRNA expression and Caspase-3 activity were higher in the HF-treated lymphocytes relative to the control group. Apoptosis could be as a possible mechanism by which Echinococcus granulosus overwhelms host defenses

Serum 25 hydroxy vitamin d3 and laboratory risk markers of cardiovascular diseases in type 2 diabetic patients

Vitamin D deficiency is prevalent worldwide. Low 25 hydroxyvitamin D3 concentration is inversely associated with type 2 diabetes, metabolic syndrome, insulin resistance, and probably cardiovascular disease. The objective of this study was to evaluate of association between vitamin D deficiency and cardiovascular risk factors among diabetic patients. This cross-sectional study, which investigated 119 type 2 diabetes patients, was conducted in Mashhad between December 2007 and March 2008. Coronary, cerebrovascular and peripheral vascular diseases in the subjects were confirmed, and blood biochemical parameters, including laboratory risk markers of cardiovascular disease were determined. Serum 25 [OH] D was measured during winter to determine the correlation between vitamin D deficiency and cardiovascular prevalent and the laboratory variables. Mean patient age was 55.3 +/- 11.2 years. Mean 25 [OH] D concentration was 32.4+21.6 ng/ml. Prevalence of vitamin D level / deficiency D was 26.1% among diabetic patients, difference with control group not being significant [P=0.12]. Overall, 36 [30.3%] patients were positive for coronary vascular disease [CVD]. The correlation between hypovitaminosis D and CVD was not significant [p=0.11]. Patients with vitamin D deficiency had significant differences in body mass index [P=0.003], metabolic syndrome [P=0.05], high sensitive CRP [P=0.009], microalbuminuria [P=0.04], and glumerolar filtration rate [P=0.02] compared with patients with sufficient vitamin D; FBS, HbAiC, lipid profiles, homocysteine, uric acid and insulin resistance were not related to vitamin D deficiency. Results showed an association between hypovitaminous D and coronary risk markers indicating the importance of this vitamin in cardiovascular health

[ study of effects of insulin and ascorbic acid on Bcl-2 family expression in hippocampus of streptozotocin -induced diabetic rats]

Diabetes is a metabolic disorder that has been shown to adversely affect both the central and peripheral nervous system by increasing basal neuronal apoptosis. Since Bcl-2 protein family is considered to play a key role in the regulation of apoptosis, in the present study we have examined the effects of insulin and ascorbic acid on expression of Bcl-2 family members including Bax [pro-apoptotic] and Bcl-2 and Bcl-x[L] [anti-apoptotic] on hippocampus of STZ-induced diabetic rats. Five groups of six Wistar rats including one control group [C] and four diabetic groups [D, I, AA and I+AA] were used in this study. Diabetes was induced by injection of 60 mg/kg STZ [IP]. After six weeks, rats in group I were treated with insulin [4-6 U/kg/day Sc], rats in group AA were treated with ascorbic acid [200 mg/kg/day IP] and rats in group I+AA were treated with equal dosage of both insulin and ascorbic acid for two weeks. Rats in group D were treated with normal saline and considered as diabetic control group. Two weeks after treatment, expression of Bcl-2, Bcl-x[L] and Bax genes were measured at both mRNA and protein levels. In diabetic control rats [group D], Bax increased whereas Bcl-2 and Bcl-x[L] decreased at both mRNA and protein levels compared to group C [P<0.01, P<0.001 respectively]. Interestingly, treatment with insulin [group I], ascorbic acid [group AA] and insulin plus ascorbic acid [group I+AA] could reverse these changes both at mRNA and protein levels [p<0.001 for I and AA+I groups, p<0.05 [Bcl-2] and p<0.01 [Bcl-x[L]] for AA group]. It is concluded that insulin and ascorbic acid alone or together can inhibit apoptosis in STZ-induced diabetic rats' hippocampus through increasing the ratio of Bcl-2/Bax and Bcl-x[L]/Bax expressions. We suggest that inhibition of apoptosis may prevent cognitive dysfunctions induced by hippocampal damage in diabetic patients as well. In addition, further experimental studies will need to be performed to confirm such effects

[ changes of stromal chondroitine sulfate in BPH and prostate adenocarcinoma]

Despite many studies, the base of formation of hyperplasia and malignancy is unknown. Prostate cancer is the second most common malignancy in men all over the world. The extracellular matrix [ECM] contains a number of macromolecules that may influence normal and neoplastic growth of tissues. The pericellular glycosaminoglycan chondroitin sulfate [GAG/Ch.s] has been ascribed roles in cell-cell adhesion, cellular matrix adhesion, cellular migration and embryological morphogenesis. In this research changes of concentration of ch.s in BPH and prostate adenocarcinoma studied. Formalin-fixed samples of BPH and adenocarcinoma were taken from patients by surgery [prostatectomy]. According to their histological characteristics [with H-E staining], samples were confirmed as BPH or prostate cancer by a pathologist. All samples were fixed in PBS-formalin, processed in standard laboratory protocols, and embedded in paraffin wax. Samples were sectioned in each 5 mm. Then critical electrolyte concentration [CEC] alcian blue staining [ph=5.8] was performed. The concentration of GAG/chondroitin sulfate in ECM was significantly higher in adenocarcinoma as compared to BPH especially in the stromal-epithelial interface. In BPH, luminal surfaces were moderately stained of ch.s but poorly stained of the adjacent fibromuscular stromal tissues or were not stained at all. In adenocarcinoma chs revealed an intense pricellular-staining pattern, the most intense staining was observed in the base membrane. The changes of concentration of GAG/ ch.s between BPH and adenocarcinoma were totally different [and specific]. The increased ch.s may be a response or requirement for epithelial growth or cell division. Therefore, the intensity of GAG/Ch.s in prostate stroma may be a useful biomarker of disease progression in prostate cancer

[Melon allergy and allergenic cross reactivity of melon with other allergens]

Allergenic reaction to melon, Cucumis melo [belongs to Cucurbitaceae family], has been reported in some allergic patients. Oral allergy syndrome was the most common clinical features associated with melon allergy. This study was aimed to confirm allergenicity of Mashadi melon, identify allergenic protein[s] of melon and allergenic cross reactivity of melon with other allergens. Prick test was performed with the extract of different parts of melon [Peel, pulp and loose layer on pulp] on the 35 patients who suffered from allergic symptoms after the ingestion of melon. Total IgE and specific IgE to melon %ere nieasurcd in 21 sera from patients with positive sicin prick test and 15 healthy controls' sera by means of ELISA. The IgE reactive protein of melon extract was detected by westeni blotting, using the 13 patient's sera [with high levels of IgE]. ELISA inhibition carried out in order to detect cross-reactivity between melon and kiwi, banana, Cynodon dactylon and Poa pratensis. Clinical reactions to melon were oral allergy syndrome 61% [immediate oral itching with or without angioedema of the lips and oral mucosa], rhinitis 38%, itching 19% and gastrointestinal symptoms 4.8%. Twenty one of the 35 patients showed positive skin prick test [SPT] to loose layer on pulp. Three patients also showed reaction to pulp and loose layer. Increased specific IgE levels to melon were observed in 18 patients with positive SPT to melon extract. Inhibition experiments showed a strong cross-reactivity of melon specific IgE with two species of ragweed pollen, especially with Cynodon doctylon, but banana and kiwi extract did not inhibit melon specific IgE in inhibition ELISA method. Immunoblot analyses of aqueous protein extract from melon were showed an IgE-binding protein of - 14.4 kDa with 8 of 12 melon-allergic patients' sera. In conclusion, we confirmed IgE-mediated hypersensitivity to melon with common clinical feature of oral allergy syndrome [OAS] and presence of an IgE-binding protein of - 14.4 kDa in melon extract. These findings suggest that main allergen of melon could be profilin

[Anti-D quantification by an enzyme-linked antiglobulin test: A comparison study of two methods]

The quantification of anti-red blood cell antibodies routinely performed by standard direct antiglobulin test [DAT], but there are some limits to its sensitivity and the end-point evaluation is very subjective in this method. Recently, a modified enzyme-linked immunosorbent assay [ELISA] has been applied to detect anti-red blood cell antibodies. In this method, the end-point is determined by enzyme activity of an enzyme conjugated anti-human antibodies. The enzyme converts a soluble substrate to a soluble colored product, which is proportional to amount of primary antibodies. This technique has been called enzyme linked antiglobulin test [ELAT]. In this study, we tried to evaluate the analytical parameters of this technique and compare with conventional DAT method. Our results showed some modification of the previously described ELAT technique makes it feasible and simpler for the routine applications. This modified method was reproducible with the mean coefficient variation of 9.08 and 5.21 for between days and within day assays and found to be at least sixteen times more sensitive than DAT method. Anti-D measurement showed an acceptable correlation [R 0.88] between ELAT and DAT methods. Therefore, this method could be useful for more precise monitoring of allo-immunized mothers and patients with autoimmune hemolytic anti-D anemia and provide an alternative method for assessing anti-D activity of specific total IgG and IgG subclasses preparation