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Abstract Background Leishmaniasis as a neglected tropical disease (NTD) is caused by the inoculation of Leishmania parasites via the bite of phlebotomine sand flies. After an infected bite, a series of innate and adaptive immune responses occurs, among which neutrophils can be mentioned as the initiators. Among the multiple functions of these fighting cells, neutrophil extracellular traps (NETs) were studied in the presence of Leishmania major promastigotes and salivary gland homogenates (SGH) of Phlebotomus papatasi alone, and in combination to mimic natural conditions of transmission. Material & methods The effect of L. major and SGH on NETs formation was studied in three different groups: neutrophils + SGH (NS), neutrophils + L. major (NL), neutrophils + L. major + SGH (NLS) along with negative and positive controls in 2, 4 and 6 h post-incubation. Different microscopic methods were used to visualize NETs comprising: fluorescence microscopy by Acridine Orange/ Ethidium Bromide staining, optical microscopy by Giemsa staining and scanning electron microscopy. In addition, the expression level of three different genes NE, MPO and MMP9 was evaluated by Real-Time PCR. Results All three microscopical methods revealed similar results, as in NS group, chromatin extrusion as a sign of NETosis, was not very evident in each three time points; but, in NL and especially NLS group, more NETosis was observed and the interaction between neutrophils and promastigotes in NL and also with saliva in NLS group, gradually increased over times. Real-time reveals that, the expression of MPO, NE and MMP9 genes increased during 2 and 4 h after exposure, and then decreased at 6 h in most groups. Conclusion Hence, it was determined that the simultaneous presence of parasite and saliva in NLS group has a greater impact on the formation of NETs compared to NL and NS groups.

Premature ovarian insufficiency (POI) has far-reaching consequences on women's life quality. Due to the lack of full recognition of the etiology and complexity of this disease, there is no appropriate treatment for infected patients. Recently, stem cell therapy has attracted the attention of regenerative medicine scholars and offered promising outcomes for POI patients. Several kinds of stem cells, such as embryonic stem cells (ESCs), mesenchymal stem cells (MSCs), and induced pluripotent stem cells (iPSCs) have been used for the treatment of ovarian diseases. However, their potential protective mechanisms are still unknown. Undoubtedly, a better understanding of the therapeutic molecular and cellular mechanisms of stem cells will address uncover strategies to increase their clinical application for multiple disorders such as POI. This paper describes a detailed account of the potential properties of different types of stem cells and provides a comprehensive review of their protective mechanisms, particularly MSC, in POI disorder. In addition, ongoing challenges and several strategies to improve the efficacy of MSC in clinical use are addressed. Therefore, this review will provide proof-of-concept for further clinical application of stem cells in POI.

Background: Coronavirus disease 2019 (COVID-19) is an infectious disease that has surrounded the world caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The disease is usually onset with symptoms like fever, cough, fatigue, respiratory problems, and loss of smell and taste. The majority of COVID-19 patients have mild or no symptoms, but a few demonstrate acute respiratory problems (ARDS) that can be life-threatening. Materials and Methods: Authors searched English published articles in local and international journals over the period 2000 to 2022 using several databases including Scopus, PubMed, Scholar, and Science Direct. Then, the relevant articles were revised. During this period, different articles have been published, but we tried to choose and review articles that introduced effective data. Results: Some people show symptoms long after their negative PCR test called post-COVID-19 syndrome, which studies showed can last more than 12 weeks after infection. Other than the complications patients confront amid the period of COVID-19 infection, there is an accumulation of evidence regarding the delayed complications of COVID-19, including auto-immune outbreaks such as multisystem inflammatory syndrome (MIS), idiopathic thrombocytopenic purpura (ITP), Guillain-Barre syndrome, Miller-Fisher syndrome, Autoimmune hemolytic anemia (AIHA), Autoimmune thyroid disease and also COVID-19 associated coagulopathies, have received remarkable attention since the early months of the pandemic. Microbiome changes in the gut and nasopharynx of patients with COVID-19 affect the severity of the disease, furthermore, some genes inherited from Neanderthals increase the severity of COVID-19. Conclusion: COVID-19 infection, along with the immune suppression mechanism, has the potential to evoke destructive inflammation in the host. Clarifying the pathophysiology of the COVID‐19 injuries to the host could help to develop appropriate treatment.

Background: The great gerbil (Rhombomys opimus), is widely distributed in Asia and is a natural reservoir for zoonotic cutaneous leishmaniasis in many endemic areas, as well as Iran. Materials and Methods: In this study, infection to Leishmania species was investigated by two methods, parasitological and molecular survey, in the small number of R. opimus collected from Jovain, a Zoonotic Cutaneous Leishmaniasis (ZCL) focus located in North East of Iran. Results: Parasitological observation showed infection in only one of five rodents. But, ITS2-Nested-PCR revealed Leishmania infection in three out of 5 gerbils, including the parasitological positive one. Based on the PCR amplified size, two cases of infections were Leishmania major and one Leishmania turanica, their sequences are accessible in GenBank. The results of sequence analysis were consistent with the results obtained based on the size of the PCR. Conclusion: These findings re-confirm the important role of R. opimus in the natural circulation of Leishmania spp and indicate the need to be concerned about the disease in the study area.

Epidemiological and clinical studies have indicated an association between particulate matter (PM) exposure and acute and chronic pulmonary inflammation, which may be registered as increased mortality and morbidity. Despite the increasing evidence, the pathophysiology mechanism of these PMs is still not fully characterised. Pulmonary alveolar macrophages (PAMs), as a predominant cell in the lung, play a critically important role in these pathological mechanisms. Toxin exposure triggers events associated with macrophage activation, including oxidative stress, acute damage, tissue disruption, remodelling and fibrosis. Targeting macrophage may potentially be employed to treat these types of lung inflammation without affecting the natural immune response to bacterial infections. Biological toxins, their sources of exposure, physical and other properties, and their effects on the individuals are summarised in this article. Inhaled particulates from air pollution and toxic gases containing chemicals can interact with alveolar epithelial cells and immune cells in the airways. PAMs can sense ambient pollutants and be stimulated, triggering cellular signalling pathways. These cells are highly adaptable and can change their function and phenotype in response to inhaled agents. PAMs also have the ability to polarise and undergo plasticity in response to tissue damage, while maintaining resistance to exposure to inhaled agents.

Abstract Background Acinetobacter baumannii is an opportunistic and antibiotic-resistant pathogen that predominantly causes nosocomial infections. There is urgent need for development nonantibiotic-based treatment strategies. We developed a novel monoclonal antibody (mAb) against a peptide of conserved outer membrane protein A (OmpA) and evaluated its reactivity with different pulsotypes of A. baumannii. Methods Peptide derived from A. baumannii OmpA was conjugated to keyhole limpet hemocyanin and injected into BALB/c mice. Splenocytes of immunized mice were fused with SP2/0 myeloma cells followed by selection of antibody-producing hybridoma cells. After screening of different hybridoma colonies by ELISA, one monoclone was selected as 3F10-C9 and the antibody was tested for reaction with five different Acinetobacter pulsotypes that were resistant to carbapenem antibiotics. The affinity constant was measured by ELISA. The ELISA, western blotting, indirect immunofluorescence (IFA), and in vitro opsonophagocytosis assays were used to evaluate the reactivity of generated mAb. Results The anti-OmpA antibody reacted with the immunizing peptide and had a high affinity (1.94 × 10−9 M) for its antigen in the ELISA. Specific binding of mAb to OmpA was confirmed in Western blot. IFA assays revealed that mAb recognized specific OmpA on the pulsotypes. Opsonophagocytosis assays showed that the mAb increased the bactericidal activity of macrophage cells. The antibody function was higher in the presence of serum complement. Conclusions The peptide-based mAb demonstrated optimal performance in laboratory experiments which may be appropriate in investigation on OmpA in Acinetobacter pathogenesis and development of passive immunization as a novel therapeutic approach.

Extensively drug-resistant Acinetobacter baumannii is considered one of the most dangerous threats to global health, requiring novel therapeutic interventions. The outer membrane protein A (OmpA) is an immunogenic agent that triggers immune responses. The current study evaluated serum antibody levels against previously determined immunogenic OmpA peptides from A baumannii in ICU staff. Serum samples were collected from 62 ICU staff members (representing the exposed group), healthy controls (representing the nonexposed group), and patients with systemic lupus erythematosus (SLE) (as controls for nonspecific antibody reactions). After excluding the cross-reactive antibodies via Escherichia coli lysate pretreatment, all the samples were assessed in the vicinity of A baumannii lysate by enzyme-linked immunosorbent assay (ELISA). All the positive samples were assessed for interaction with previously designed and selected peptides using ELISA. The protective potential of positive serum antibodies was surveyed in vitro using an opsonophagocytic study. The most antibody positive samples against one of the dominant peptides were determined in the ICU personnel (75%). SLE serum samples did not react with candidate peptides. The strongest positive reaction was observed in serum treatment with one of the OmpA peptides (No. 5) with significant differences compared to other designed peptides. Our findings showed that ICU samples have substantially higher antibody levels than the nonexposed group; Positive samples show strong results in the opsonophagocytosiis assay. This study demonstrates A baumannii colonization at human mucosal surfaces, especially in exposed healthy workers. Novel OmpA-derived peptides could be used to identify immunogenic vaccine candidates. Therefore, more studies are needed before this peptide and antibody levels are used in diagnosis, prevention, or treatment.

Abstract Background: Premature ovarian insufficiency (POI) affects about 1% of women of reproductive ages (15-45 yr), with no curative treatment. Objective: We aimed to present a rat model of POI using a D-galactose enriched diet. Materials and Methods: In a pilot study, 4 pregnant Wistar rats were divided into 4 groups; 3 groups were fed galactose-enriched diets at days 3-15 of pregnancy (G1); on the 3 rd day of pregnancy to parturition (G2), and the 3 rd day of pregnancy until the end of the weaning period (G3). Also, group 4, as the control group (G0), was fed standard pellets during the study. After confirming the lack of adverse effects of dieting with galactose in terms of offsprings' birth weight, we performed our study designed the same as the pilot study. A total of 40 pregnant Wistar rats were randomly divided into 4 groups. Ovarian histology, reproductive hormones, and immunological characteristics of the female offspring were examined in all experimental groups and compared. Results: The pilot study revealed no significant differences in the birth weight of the offspring of the 4 study groups (p = 0.96). The ovarian index in the female offspring of those with a gal-exposed diet was significantly lower than that of the control group offspring (p < 0.01). Conclusion: As the birth weights of the offspring of our experimental and control groups were similar, it can be concluded that the reduction of ovarian follicles after prenatal exposure to D-galactose is due to the ovotoxicity of galactose. The results of our final study will provide more information about the rat POI model induced by prenatal exposure to D-galactose.

Fibrosing pneumonia (FP) is classified into usual interstitial pneumonia (UIP) and nonspecific interstitial pneumonia (NSIP), each having its own etiology and prognosis. Both types of FP are progressive and chronic conditions with distinct etiologies. Cytokines and inflammatory mediators play critical roles in the pathogenesis of FP. Among them, the role of transforming growth factor beta-1 (TGF-β1) and modulators triggering fibrosis are not well understood. In this study, the expression of triggering receptor expressed on myeloid cells-1 (TREM-1) as a stimulator for the production of TGF-β1 and also CD4+CD25+Foxp3+ regulatory cells were investigted in FP patients. Sixteen UIP, 14 NSIP and 4 pulmonary fibrosis following Mycobacterium tuberculosis (TB) infection patients, were compared with 12 healthy controls. The frequency of blood CD14+TGF-β1+ and CD14+TREM1+-gated monocytes and CD4+CD25+Foxp3+ regulatory T cells (Treg), as well as the plasma levels of TGF-β1 and IL‑10 were measured. Fibrosis patients compared to healthy controls had a greater frequency of CD14+TGF-β1+ [15.9 (0.2-88.2) vs. 0.6 (0.2-11.0)] and CD14+TREM1+ [21.1 (2.3-91.2) vs. 10.3 (3.1-28.6)]-gated monocytes, and CD4+CD25+Foxp3+ [1.2 (0.3-3.6) vs. 0.2 (0.1-0.4)]-gated lymphocytes. Plasma TGF-β1 were also significantly increased in patients with fibrosis compared to healthy controls [9316.2 (±5554.4) vs. 3787.5 (±2255.6)]. These results confirm the importance of TGF-β1 and TREM1 in pulmonary fibrosis. It seems that this reciprocal cycle in healthy people is modulated by the production of IL‑10 by Treg cells, thus limiting fibrosis, as observed in patients following TB infection. Further investigations are recommended to evaluate possible immunomodulatory mechanisms defects in pulmonary fibrosis.

Objective(s): One of the causes of human and animal zoonotic infections is Brucella melitensis, which is transmitted to humans through dairy products. It seems for prevention of human infection we might protect the livestock by an efficient protein as a vaccine candidate. For this purpose, the use of immunogenic proteins of bacteria is able to create immunity the same as the traditional vaccines.Materials and Methods: In this study, by finding the immunogenic antigens of this bacterium by 2-dimensional gel electrophoresis and MALDI-TOF methods and also the proteins reported in other studies, we found the epitopes of the bacterial antigenic determinants in silico. Nineteen peptides of T and B epitopes were selected. They were ligated with linkers and after gene synthesis, the designed polypeptide was expressed in Escherichia coli BL21. The purified recombinant MEL protein mixed with chitin was injected subcutaneously into three 300 g male guinea pigs three times. Also, PBS control and Rev.1 commercial vaccine groups were considered. Results: The results show that MEL polypeptide is equal to the Rev.1 vaccine in stimulating secretion of IFNγ and IL2 and specific IgG. High levels of IL-2 emphasize the activation of the cellular immunity, and in particular comparison of PI in guinea pig’s spleen cells treated with recombinant MEL protein on days 0 and 5 show that it has significant proliferation compared with PBS unstimulated cells.Conclusion: This recombinant protein could be a subunit protein with sufficient efficiency in stimulating the humoral and cellular-mediated immune system against B. melitansis.

Objectives: This study investigates the therapeutic effect of vitamin C on the development of endometrial lesions and fecundity disorders in the ovarian induction model of mouse endometriosis. Methods: Ovarian endometriosis was surgically induced in 14 NMRI female mice (treatment group, N = 7) and (control group, N = 7). Three days after the second surgery (to assess endometriotic implant), the mice were randomized into two intervention groups: control (placebo) and treatment (50 mg/kg vitamin C every two days orally for four weeks) groups. In the oestrus phase, the mice were sacrificed. In macroscopic assessment, endometriotic implants were evaluated in size, volume, weight, growth score and adhesion score. The microscopic assessment examined the ovarian tissue (the number of antral follicles, corpus luteum and atretic follicles) and endometriotic lesion (histologic and trichrome fibrosis scores). Results: Post-treatment implant volume, growth score, adhesion extent score and adhesion severity score were significantly lower in the treatment group (vitamin C) in comparison with the control group (placebo) (p

Background: Treatment of leishmaniasis is a challenge due to problems such as high price and dose of the drug, drug resistance and side effects. Objective: The study aims to introduce plants compounds, which their antileishmanial effects were approved in vivo conditions. Methods: This study as a review article was performed by searching the keywords of “medicinal plants with in vivo antileishmanial effects, nanocarrier, clinical trials, and mechanism of action” in the well-known databases to 2018. In this study, 14 medicinal plant compounds with antileishmanial effects were reviewed and mechanism of action and their in vivo therapeutic effect were evaluated. Results: It was found that while some of these compounds had low antileishmanial effects, their efficacy against leishmaniasis was significantly increased through loading into nanocarriers. Conclusion: This study indicated that active component of medicinal plants especially along with nano carriers can be of interest for the treatment of Leishmania.

Background: The amniotic membrane plays an important role in maintaining a healthy pregnancy. The main population cells from amniotic membrane include human amnion epithelial cells (hAECs) which have been shown to possess immunomodulatory properties.Objective: The proximity of hAECs with monocyte leads to the generation of tollerogenic dendritic cells.Materials and Methods: hAECs were obtained from normal pregnancy. Peripheral blood monocytes were isolated by anti-CD14 MACS method. Co-cultures of monocytes and hAECs were established in Transwell chambers supplemented with granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4) in the absence and presence of lipopolysaccharide (LPS) to produce immature and mature DCs, respectively. Immunophenotyping of the obtained DCs was done through flow cytometry and the production of cytokines was measured by ELISA. Mixed leukocyte Reaction (MLR) was also performed for the functional assessment of DCs.Results: Immunophenotyping of [hAECs - Immature DC (iDC)] and [hAECs - iDC] + LPS cells revealed that the expression of CD1a, CD80, CD86, CD40, HLA-DR, and CD83 markers showed no significant difference as compared with the control group (iDCs and mDCs alone). In the [hAECs-iDCs] + LPS cells, the percentage of CD14 cells at the ratio of 1:2.5 showed significant differences compared to the control group. The production of IL-10 and IL-12 showed no significant difference in any of the cultures as compared to the control groups. Also, co-cultured DCs did not inhibit proliferation of lymphocyte.Conclusion: Our findings show that factors secreted from cultured hAECs are unable to generate of tollerogenic dendritic cells. To achieve a better understanding of other mechanisms more investigations are needed.

Background and Objectives: Immunization is a promising strategy to combat against the life-threatening infections by Multi Drug Resistance Acinetobacter baumannii. In this study, we directed to design and evaluate the efficacy of a recombinant multi-epitope protein against this pathogen. Materials and Methods: Epitopes prediction was performed for candidate proteins OmpA and BAM complex (BamA, BamB, BamC, BamD, BamE) from A. baumannii, using immune-informatics tools with high affinity for the human HLA alleles. After expression and purification of the recombinant protein, its functional activity was confirmed by interaction with positive sera. Results: Cloning and expression of the desired multi-epitopes protein were verified. Circular Dichroism study showed the secondary structure and proper refolding of the recombinant protein was achieved and matched with computational prediction. There was a significant interaction between designed protein with antibodies presented in ICU patients' and staff's sera. Conclusion: The interaction of the recombinant protein with patients' sera antibodies suggests that it may be a promising determinant protein for immunization against of MDR A. baumannii.

Nosocomial infections caused by Acinetobacter baumannii (A. baumannii) nosocomial infections caused by Acinetobacter baumannii (A. baumannii) are considered as a global serious problem in hospitalized patients because of emerging antibiotic resistance. Immunotherapy approaches are promising to prevent such infections. In our previous study, five antigenic epitopes of outer membrane protein A (OmpA), as the most dangerous virulence molecule in A. baumanii, were predicted in silico. In this study, the investigators evaluated some immunological aspects of the peptides. Five peptides were separately injected into C5BL/6 mice; then the cytokine production (interleukin-4 and interferon-gamma) of splenocytes and opsonophagocytic activity of immunized serum were assessed. To identify the protective function of the peptides, animal models of sepsis and pneumonia infections were actively and passively immunized with selected peptides and pooled sera of immunized mice, respectively. Then, survival rates of them were compared with the non-infected controls. Based on the results, activated spleen cells in P127 peptide-immunized mice exhibited an increase level of IFN-γ compared with the other experimental groups, but not about the IL-4 concentration. The results of opsonophagocytic assay revealed an appropriate killing activity of produced antibodies against A. baumannii in a dose-dependent manner. Further, the survival rates of the mice under passive immunization with the immunized sera or active immunization with P127 peptide were significantly more than those in the control group. Moreover, the survival rate of the P127 peptide immunized group was considerably higher than that among the other peptide-immunized group. In conclusion, findings indicated that peptides derived from outer membrane protein-A can be used as a promising tool for designing the epitope-based vaccines against infections caused by A. baumannii.

Abstract Background Development of a hyper-gonadotropic hypoestrogenism condition in women

Background & Objective: Ischemia and Reperfusion (I/R) lead to the production of excessive reactive oxygen species (ROS). In the present study the protective effects of the cell culture conditioned medium (CM) on sperm quality disorder induced by torsion-detorsion (T-D) in adult rat’s testis was evaluated. Materials & methods: In this study 42 adult rats were divided into seven groups. 1- Normal control group treated without surgery. 2- Laparotomy group, surgery operated without T-D. 3- T-D group, surgery operated along with T-D. 4- T-D+DMEM group, after T-D Dulbecco's Modified Eagle's Medium (DMEM) has been injected. 5-7 groups included T-D+F, T-D+M, and T-D+FM, along with T-D injected conditioned medium of fibroblast, macrophage, and fibroblast-macrophage co-culture respectively. All injections were made at a rate of 10μL once after T-D in the rete testis. After 35 days, sperm quality parameters were evaluated and obtained data were compared between groups by one-way ANOVA statistic method and Tukey test post hoc in P

The presence of ambient particulate matter (PM) poses more dangers to human health than that of other common air pollutants such as Carbon dioxide (Co2) and ozone. Epidemiologic studies show a direct correlation between PM and the risk of respiratory and cardiovascular diseases. The immune system seems to play a critical role in the process of these diseases. The main goal of this study was to investigate the effect of Tehran particulate matter in two aerodynamic diameters (PM2.5 and PM10) on alveolar macrophages (AM) from C57/BL6 mice. To evaluate the inflammatory effects of PMs, cultured alveolar, and peritoneal macrophages were treated with PM2.5 and PM10 (concentrations of 5 µg/mL and 10 µg/mL). Tumor necrosis factor-alpha (TNF-α) and IL-10 (representatives of inflammatory and anti-inflammatory cytokines, respectively) were assessed in the culture supernatant by ELISA. Expression of arginase and inducible nitric oxide synthase (iNOS) genes was carried out by quantitative real-time PCR. Different functional types of cultured alveolar macrophages (M1, M2) were also determined in this study. Our results suggest that PM2.5 induces M1 inflammatory phenotype in comparison with PM10. We found Also, an increase in TNF-α and M1-related gene expression (iNOS), as well as a decrease in both IL-10 and M2 phenotype genes (Arginase). Moreover, a reduction in phagocytic capacity and increased apoptosis function of macrophage cells were detected. PM2.5 as a major component in hydrocarbons has a considerable effect on polarizing the alveolar macrophages to an inflammatory phenotype and eliciting lung inflammation in mice.

INTRODUCTION:Irradiation can induce multiple inhibitory and stimulatory effects on the immune system. In recent studies, it has been noted that administration of radiation with various doses and fractionation plans may influence on immune responses in microenvironment of tumor. But in radiobiology, the Biologically Effective Dose (BED) formula has been designed for calculating isoeffect doses in different regimens of daily clinical practice. In other words, BED has also been used to predict the effects of fractionation schedules on tumor cells. METHODS:In our study, three different regimens with BEDs of 40 gray (Gy) were analyzed in BALB/c mice. These included conventional fractionated radiotherapy (RT) (3Gyx10), high-dose hypofractionated RT (10Gyx2), and single ablative high-dose RT (15Gyx1). RESULTS:As BED predicts, all three similarly decreased tumor volumes and increased survival times relative to controls, but after high dose exposure in ablative group, the expression of IFNγ was increased following high infiltration of CD8 cells into the tumor microenvironment. When anti-PDL-1 was combined with RT, single ablative high-dose radiation enhanced antitumor activity by increasing IFNγ in tumors and CD8+ tumor-infiltrating lymphocytes; as a result, this combining therapy had enhanced antitumor activity and lead to control tumor volume effectively and improve significantly survival rate and finally the recurrence of tumor was not observed. CONCLUSION:Results show distinct radiation doses and fractionation schemes with same BED have different immunogenic response and these findings can provide data helping to design regimens of radiation combined with immune checkpoint blockers (ICBs).

Abstract Objective Human amniotic epithelial cells (hAECs) are a novel source of stem cells and have immunomodulatory effects on both the innate and adoptive immune system. hAECs can differentiate into multiple cell lineages that make them a suitable cell source for regenerative medicine. These cells express multiple toll-like receptors (TLRs) and respond to various TLR ligands. This study aimed to evaluate the effect of lipopolysaccharide (LPS), a TLR4 ligand, on the level of immunomodulatory and immunostimulatory factors of hAECs. Results Our results indicated that LPS had the ability to up-regulate the expression of prostaglandin E2 synthase and transforming growth factor-beta1 in hAECs. However, there was no change in the level of interleukin-1beta, interleukin-6 and interleukin-10 in hAECs when were stimulated with LPS. In addition, we observed tumor necrosis factor-alpha was only expressed at very low level in some of hAECs samples which its expression was independent of the effects of LPS.

Acinetobacter baumannii is a Gram-negative bacterium that has recently been identified as a leading nosocomial pathogen. Infections by this pathogen result in significant mortality due to antibiotic resistance. An effective vaccine would help alleviate the burden of disease incurred by this pathogen; however, there are currently no licensed vaccines offering protection against Acinetobacter baumannii infection. In this study, considering the fact that outer membrane protein A is one of the most promising vaccine candidates, we predicted T cell and B cell epitopes on this protein using sequence-based epitope prediction tools and determined whether or not mice immunized with these peptides induce an immune response. We selected consensus epitopes including five peptides in different tools with the highest score. 48 female C5BL/6 SPF injected subcutaneously with the peptides (peptide1 to peptide 5 separately) in 100 μL of the solution and sham groups received adjuvant and PBS alone on the same schedule: on day 0 (primary dose) and two booster doses were administered on days 14 and 28. At the end of time, animals euthanized by Isoflurane, and collected sera for assessment of specific antibodies against each peptide by ELISA (Enzyme-linked immunosorbent assay). Immunization of mice showed one of the novel synthetic peptides (peptide 1 (24-50 amino acids)) elicited immune responses. We conclude to combine theoretical methods of epitope prediction and evaluating the potential of immunogenicity for developing vaccines is important.

Silencing of gene expression by siRNA (small interfering RNA) is a powerful approach used to study the genetic analysis and functional roles of mammalian genes. There is at present no report about the effects of mammalian two-hybrid system plasmids delivery of sense and antisense strands. The leishmania pteridine reductase 1 (PTR1) gene was cloned as sense and antisense strands into mammalian two hybrid system plasmids. The constructs were transfected into human blood macrophages on the basis of eight experimental groups. (Antisense strand ± LPS, sense strand ± LPS, dsRNA ± LPS, negative control ± LPS). After 24 hours, cytokines production was assessed with ELISA. Transfection of sense and antisense strand RNA into monocyte-derived macrophages (MDM) was confirmed by RT-PCR. Single strands RNA expressed IL-8, IL-12, IL-1β inflammatory cytokines and dsRNA induced IL-8, IL-12 and TNF-α production in MDM. In contrast, random uptake from a mixture of two plasmids was downregulated IL-8, IL-12, IFN-γ cytokines, with a significant difference of p

Objective: To determine the preventive effects of local administration of simvastatin for postoperative intra-abdominal adhesion formation in animal model of rat.Methods: In this experimental study, 32 Wistar albino rats as the animal model of intra-abdominal adhesion formation were included. Adhesions were induced in all the animals via abrasion of the peritoneal and intestinal surface during laparotomy. Afterwards, the rats were randomly assigned to receive simvastatin (30 mg/kg body weight) as a single intraperitoneal dose at the time of laparotomy (n=16) or normal saline in same volume at the same time (n=16). At the day 21, animals were euthanized and the adhesions were quantified clinically (via repeated laparotomy) and pathologically and compared between the two groups.Results: The baseline characteristics of the animals were comparable between two study groups. Clinically, in simvastatin group, 10 rats (62.5%) did not develop any adhesion and 6 (37.5%) had first-grade adhesion; whereas in the control group, 11 (68.8%) rats had first- and 5 (31.2%) had second-grade adhesions (p

Background: Herbal medicines in compared with chemical drugs have fewer side effects and can be a good medicinal alternative. The olive includes 20 different species of the family Oleaceae, with Olea europaea as the most recognized. Several studies have shown the immunomodulating effects of olive leaf extract. This study aimed to identify the immunoregulatory effect of olive leaf Sevillana variety on interleukins 12 and 10 which resulted from the murine macrophages in vitro.Materials and Methods: In order to isolate macrophages, peritoneal macrophages BALB/C were used. To determine the cytotoxic effect of different concentrations of the olive leaf extract on macrophages, MTT assay was performed. Concentrations of 200, 100, 50, 25, 12.5, 6.25, and 3.1μg/ml in the time intervals of 12, 24, and 48 hours were evaluated. Three appropriate concentrations were selected to commence with the study of the determination of the amount of cytokines. Cell culture supernatant growth medium supernatant was collected at 12, 24, and 48 hours after adding the extract in order to examine the amount of cytokines. ELISA test was conducted using interleukins 12 and 10 measurement kits.Results: CC50 of the olive leaf extract at 12, 24, and 48 hours was 260.3, 170.5, and 150 μg/ml, respectively. According to the results, an increase in the concentration and duration of the study resulted in observable significant differences in the production of interleukins 10 and 12. As a result, the production of IL-10 and 12 experienced decreases and increases, respectively.Conclusion: It seemed probable that the olive leaf extract had the capability to increase the production of IL-12 through activation of the classic macrophages and also deactivate the regulatory macrophages with an increase in IL-12 and a decrease in IL-10. Therefore, this can strengthen the immune system of the host in the early stages of infection. The other immunomodulatory effects of olive leaf must be considered by appropriate research.

Background- CD24 is a cell adhesion molecule that has been implicated in metastatic tumor progression cells. Our aim was clarify correlation between CD24 expression and cAMP:cGMP ratio in murine colorectal cancer cell line (CT26) after using cholera toxin.Materials and Methods- The CT26 cells were cultured in microtubes for assaying cAMP and cGMP; also the cells were cultured in flasks for assaying cytoplasmic and membranous CD24 expression. The Real-Time PCR was done for cDNA that was synthesized from CT26 cells’ mRNA. Also, expression CD24 marker of cells was determined by Anti-CD24 antibody and Goat Anti-Rabbit IgG-FITC (flow cytometry).Results- The cholera toxin grew cAMP:cGMP ratio and it influenced cytoplasmic and membrane CD24 expression.Conclusion- There are indirect correlation between cAMP:cGMP ratio and CD24 expression.

Objective: Nuclear transfer-embryonic stem cells (NT-ESCs) are genetically identical to the donor’s cells; provide a renewable source of tissue for replacement, and therefore, decrease the risk of immune rejection. Trichostatin A (TSA) as a histone deacetylase inhibitor (HDACi) plays an important role in the reorganization of the genome and epigenetic changes. In this study, we examined whether TSA treatment after somatic cell nuclear transfer (SCNT) can improve the developmental rate of embryos and establishment rate of NT-ESCs line, as well as whether TSA treatment can improve histone modification in NT-ESCs lines.Materials and Methods: In this experimental study, mature oocytes were recovered from BDF1 [C57BL/6×DBA/2) F 1 mice] mice and enucleated by micromanipulator. Cumulus cells were injected into enucleated oocytes as donor. Reconstructed embryos were activated in the presence or absence of TSA and cultured for 5 days. Blastocysts were transferred on inactive mouse embryonic fibroblasts (MEF), so ESCs lines were established. ESCs markers were evaluated by reverse transcription-polymerase chain reaction (RT-PCR). Histone modifications were analyzed by enzyme linked immunosorbent assay (ELISA).Results: Result of this study showed that TSA treatment after SCNT can improve developmental rate of embryos (21.12 ± 3.56 vs. 8.08 ± 7.92), as well as establishment rate of NT-ESCs line (25 vs. 12.5). We established 6 NT-ESCs in two experimental groups, and three embryonic stem cells (ESCs) lines as control group. TSA treatment has no effect in H3K4 acetylation and H3K9 tri-methylation in ESCs.Conclusion: TSA plays a key role in the developmental rate of embryos, establishment rate of ESC lines after SCNT, and regulation of histone modification in NT-ESCs, in a manner similar to that of ESCs established from normal blastocysts.

The dorsal skin of the Iranian frog, Rana ridibanda, is associated with numerous prominent granular glands which extract their secretions in the response to stressor or invading pathogens .The secretions have broad spectrum antimicrobial effects. In this research the effect of antimicrobial skin secretions from Iranian frog (Rana ridibanda) has been examined against gram positive Methycillin Resistant Staph Aureus (MRSA), under sterile conditions. To show this effect, 1cm² of frog skin was cultured in complete tissue culture medium containing RPMI, FBS and FUNGISON for the period of 10 days. Immediately after, MRSA was exposed to frog skin secretion (medium culture) and Vancomycin. The results showed that the frog skin secretions has significant antimicrobial effect against MRSA. The range of inhibition zone for MRSA was the same as (20mm) Vancomycin in DISK method. In Minimum Inhibitory Concentration method, for MRSA, the tube 1/8 was positive.