The present work sought to investigate potential suppressive effects on mouse macrophages by in vitro treatment with clove (Syzygium aromaticum) ethanolic extracted essential oil (containing eugenol) or its water-soluble extract. Using doses (ranging from 0.001-1000 mg/ml) of each material freshly prepared in the laboratory, cell survival and production of nitric oxide (NO), tumor necrosis factor (TNF)-, interleukin (IL)-6, and IL-12 by the treated cells (that in all cases also had received LPS stimulation) were measured. Results indicated that, except at doses !100 mg/ml, viability was unaffected in all groups. NO release by LPS-stimulated macrophages was generally significantly suppressed by either material; in contrast, low (i.e. 0.001-1 mg/ml) doses of either extract class appeared to enhance NO release by non-LPS (unstimulated)-treated macrophages. Among LPS-stimulated cells, TNF release was also significantly affected by each extract; the ethanolic extract was suppressive at all doses tested, while the aqueous material was so up to 1 mg/ml and then became stimulatory. In contrast, nearly every dose of either extract appeared to stimulate IL-6 release from the LPS-treated cells. Effects on IL-12 production were overall inconsistent; in general, the ethanolic extract tended to be stimulatory of production by the LPS-treated cells. The data for the aqueous material showed no discernable pattern of effect. The results suggest that clove extracts do not have a distinct cytotoxic activity, but do impart potential anti-and pro-oxidant effects in cells, depending on their concentrations and on the activation state of the macrophages themselves at the time of exposure to the extracts. The impact of the extracts on macrophage cytokine release also displays a pattern of dose-relatedness.
Clove (Syzygium aromaticum) has been used in folk medicine in many disorders. The present work aimed to investigate effects of clove essential oil as eugenol and water soluble ingredients on mouse splenocytes. Clove extracts were harvested and in different concentrations (0.001-1000 μg/mL) were affected to splenocytes and also phytohemagglutinin (PHA = 5 μg/mL) and lipopolysaccharide (LPS = 10 μg/mL) activated splenocytes; then splenocytes proliferation assayed using the MTT ([3-(4, 5-dimethylthiazole-2-yl) -2, 5-diphenyl tetrazolium bromide]) method were done. On the culture supernatant interferon (IFN)-γ, interleukin (IL)-4, IL-10, and transforming growth factor (TGF)-β cytokines were measured. Clove ingredients (100 μg/mL and 1000 μg/mL) reduced PHA stimulated splenocytes proliferation and enhanced LPS stimulated cells expansion. Treated splenocytes showed suppression of IFN-γ release and induction of IL-4, IL-10, and TGF-β secretion (in the range of 0.1-1000 μg/mL). The results of this study suggest clove extracts could suppress the T cell cellular immunity and enhance humoral immune responses. In clove affection cytokine pattern shifted toward modulatory and Th2 responses and accelerator of humoral immunity cytokines.
In the body, there is a natural three-dimensional (3D) microenvironment in which immune cells, including dendritic cells (DC), play their functions. This study evaluated the impact of using collagen-chitosan 3D nano-scaffolds in comparisons to routine 2D culture plates on DC phenotype and functions. Bone marrow-derived DC were cultured on scaffolds and plates and then stimulated with lipopolysaccharide (LPS) or chitosan-based nanoparticles (NP) for 24 h. Thereafter, DC viability, expression of maturation markers and levels of cytokines secretion were evaluated. In another set of studies, the DC were co-cultured with allogenic T-lymphocytes in both the 2D and 3D systems and effects on DC-induction of T-lymphocyte proliferation and cytokine release were analyzed. The results indicated that CD40, CD86 and MHC II marker expression and interleukin (IL)-12, IL-6 and tumor necrosis factor (TNF)-secretion by DC were enhanced in 3D cultures in comparison to by cells maintained in the 2D states. The data also showed that DNA/chitosan NP activated DC more than LPS in the 3D system. T-Lymphocyte proliferation was induced to a greater extent by DNA/NP-treated DC when both cell types were maintained on the scaffolds. Interestingly, while DC induction of T-lymphocyte interferon (IFN)-and IL-4 release was enhanced in the 3D system (relative to controls), there was a suppression of transforming growth factor (TGF)-production; effects on IL-10 secretion were variable. The results here suggested that collagen-chitosan scaffolds could provide a proinflammatory and activator environment to perform studies to analyze effects of exogenous agents on the induction of DC maturation, NP uptake and/or cytokines release, as well as for the ability of these cells to potentially interact with other immune system cells in vitro.
Aim:Food and medicinal applications of barberry date back to 2500 years ago. This study investigates Berberis integerrima impact on lymphocytic immune responses.Materials & methods:Balb/c splenocytes were treated by 0.001–1000 μg/ml of B. integerrimaaqueous and alcoholic extracts in presence of phytohemagglutinin and lipopolysaccharide mitogens. Cell proliferation was assayed and cytokines were measured using ELISA.Results:Both extracts suppressed proliferation of phytohemagglutinin stimulated splenocytes (as T cells), while alcoholic extract induced expansion of lipopolysaccharide activated cells (as B lymphocytes) and unstimulated cells (p < 0.05). Both barberry extracts suppressed IFN-γ production (p < 0.05) and enhanced IL-4, IL-10 and TGF-β release from splenocytes (p < 0.05).Conclusion:Both extracts could suppress T-cell and enhance B-cell proliferation and shift immune responses toward Th2.
Background and Aims: Acinetobacter baumannii is an important multidrug-resistant opportunistic pathogen frequently causing various nosocomial infections and is a serious threat to burn patients. These infections are usually caused by the outbreak strains. The aim of this study was to show antibiotic resistance pattern and molecular typing of A.baumannii genes isolates collected from burn patients and also distribution of different types of burn patients. Materials and Methods: In this study, 307 different strains were detected. Totally 100 A.baumannii strain was selected in burn center of Isfahan hospital. Antibiotic resistance pattern was determined by disk diffusion method (Kirby Bauer). The presence of genes coding in antibiotic resistance were analyzed by using M-PCR method. The standard strains of Escherichia coli ATCC 25922 and A. baumannii ATCC 19606 were used as negative and positive controls. Results: The antibiotic resistance pattern for A.baumannii showed high resistance for ciprofloxacin, ceftazidime, and tetracycline with frequency of 82.5%, 75.3%, 72%, respectively. Moreover, the most sensitive antibiotics were chloramphenicol, and nitrofurantoin with the resistance frequency of 3.9% and 2.8%. CITM (91.1%) was the highest detected gene. Conclusions: High prevalence of antibiotic resistance pattern among A.baumannii isolated from burn center hospitals indicates the important role of multidrug resistant isolates.
Background and Aims: Ficolins are proteins that bind to carbohydrates, act as opsonins and play an important role in innate immunity. Polymorphism in ficolin-3 gene (FCN3) can lead to complement deficiency and increase the risk of some disorders such as diabetes. The aim of this study was to investigate the frequency of FCN3 + 1637delC as a single nucleotide polymorphism (SNP) in this gene in healthy and diabetic subjects of Iran. Materials and Methods: Blood was taken from 36 diabetics and 37 healthy subjects who had referred to the Iranian Blood Transfusion Organization. Blood sugar was analyzed using a calorimetric method. After DNA extraction using salting out method, polymerase chain reaction (PCR) was carried out and the restriction fragment length polymorphism (RFLP) method was accomplished using ApaI restriction enzyme. Consequently, the resulted fragments were evaluated using electrophoresis on 2% L-agarose gel. Results: Evaluation of the results indicated that the heterozygote form of SNP FCN3 + 1637delC was seen in three samples (8.1%) of the studied healthy subjects and in two samples (5.6%) of the diabetic individuals. Besides, the homozygous form of the mutation was not seen in the studied healthy and diabetic subjects. Conclusion: Results of this study showed that FCN3 variant of SNP FCN3 + 1637delC was not considered as a risk factor for type 2 diabetes mellitus (T2DM) in Iranian subjects.
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