Benefits of conditioned medium of nicotine-pulsed mesenchymal stem cells in experimental autoimmune hepatitis.

Previous data indicated that nicotine could modulate the immune regulatory potential of mesenchymal stem cells (MSCs). Currently, we intend to assess the effects of a conditioned medium of nicotine-pulsed mesenchymal stem cells in the experimental model of autoimmune hepatitis (AIH). Bone marrow-derived MSCs pulsed with 0,.1,.5, or 1 µM nicotine until the cells reached 90% confluency. Correspondent to in vitro results, the least effective concentration of nicotine that led to an anti-inflammatory environment by the MSC-conditioned medium was 0.5 µM. The murine model of AIH induced by Intravenous injection Concanavalin A (ConA). Mice were allocated to pretreatment (Concomitant treatment with ConA administration) or treatment groups and received un-pulsed MSC-conditioned medium (CM) or conditioned medium of nicotine (0.5 µM)-pulsed MSCs (CMN). The levels of ALT, AST, MPO, TNF-α, IFN-γ, and IL-6 were the highest in the ConA group than in the other groups. Pretreatment or treatment with the CMN caused a significant reduction in hepatic enzymes and inflammatory cytokines compared to pretreatment or treatment with CM. Both CM or CMN significantly decreased the numbers of activated TCD4+ and TCD8+ in the blood. More importantly, pre-treatment or treatment with CMN caused a better improvement in the histopathological appearance than pre-treatment or treatment with CM. The results of this study show that CMN rapidly controls the AIH mouse model, and therefore it may be considered as a new therapeutic approach for the treatment of AIH patients.

Benefits of bone marrow-derived mesenchymal stem cells primed with estradiol in alleviating collagen-induced arthritis.

Objectives: To investigate the effects of the oestradiol (ES) pulsed bone marrow-derived mesenchymal stem cells (BM-MSC) to treat adjuvant-induced arthritis in Wistar rats. Materials and Methods: BM-MSCs were pulsed with ES (0, 10,100, and 1000 nM) for 24 hr. RA was induced by collagen and Freund's Complete Adjuvant into the base of the tail of Wistar rats. Results: The least effective concentration of ES that can promote potent anti-inflammatory properties in the MSC population is 100 nM. At this concentration, ES increases the inhibition of the polyclonal T lymphocyte proliferation, production of IDO, IL-10, Nitric oxide, and TGF-ß, and expression of CXCR4 and CCR2 mRNA in the MSC population. Accordingly, the RA rats were treated with 2×106 MSCs or ES-pulsed MSCs (100 nM) on day 10 when all animals had developed signs of RA. ES-pulsed BM-MSCs reduced the severity of RA more profoundly than treatment with BM-MScs alone. The ability of ES-pulsed BM-MSCs to reduce symptoms and RA markers like CRP, RF, and nitric oxide was comparable to that of prednisolone. Prednisolone was more successful in reducing inflammatory cytokines than treatment with ES-pulsed BM-MSCs. ES-pulsed BM-MSCs were more successful in increasing anti-inflammatory cytokines than treatment with Prednisolone. The ability of ES-pulsed BM-MSCs to decrease the level of nitric oxide was comparable to that of prednisolone. Conclusion: ES-pulsed BM-MSCs may be a helpful strategy in RA control.

Effect of Nicotine on Immune System Function.

As a parasympathetic alkaloid and the main substance in cigarette smoke, nicotine modulates the immune system, inhibits innate and acquired immunity and is used in treating many autoimmune diseases. It often stimulates the α7 receptor and causes an anti-inflammatory state in the body. This study is designed to evaluate the role of nicotine treatment on immune system. The results showed that nicotine affects many cells in immune system, alters the downstream intracellular mechanisms and changes lymphocytes polarization. This substance alters TLRs and STATs gene expression and thus changes in the innate immune system. All these events inhibit the secretion of pro-inflammatory cytokines and chemokines which increase angiogenesis and metastasis and exacerbates tumors due to increasing survival and cell growth. Nicotine can aggravate tumors in cancer patients, with many positive effects observed in the treating autoimmune disease, Nicotine treatment function in different conditions depends on factors such as concentration, how it is employed, treatment duration and other conditions such as body conditions affecting the immune system, hence, further studies and review of all conditions are required.

Combined atorvastatin and pentoxifylline in ameliorating inflammation induced by complete Freund's adjuvant.

Injection of complete Freund's adjuvant (CFA) into one of the footpads of Wistar rats promotes swelling in all the footpads (including non-injected footpads) in the next few days, indicating a complicated immunological reaction and autoimmune arthritis. This survey was performed to assess the synergistic benefits of combined atorvastatin and pentoxifylline for treating arthritis induced by CFA. Therapeutic regimes began on day 12 post-challenge when all the rats recorded an arthritis index of more than one and continued throughout the investigation until day 32. Treatment with combined atorvastatin and pentoxifylline at half doses led to synergistic benefits causing the regression of clinical and radiological appearance of arthritis in non-injected paws, which was more favorable than treatment with any medication alone at full doses. Moreover, the combined treatment led to a reduction in some inflammatory biochemical parameters, such as myeloperoxidase, nitric oxide, and C-reactive protein, which was more pronounced than those of the treatment with each medication alone. The mRNA expression of IL-1ß and TNF-α in the rat toe area was significantly decreased by combination therapy, and this reduction was more significant than that of monotherapy. The ratios of RORγc/T-bet, RORγc/GATA-3, RoRγc/Foxp3, T-bet/GATA-3, and T-bet/Foxp3 expression showed a further decrease in the combined treated group compared to other groups. Interestingly, combination therapy did not reduce T lymphocyte proliferation to a level lower than healthy rats. Overall, the combination of atorvastatin and pentoxifylline possesses immunomodulatory synergistic benefits, and this approach may be an impressive strategy to manage complicated inflammation.

Transplantation of decellularised human amniotic membranes seeded with mesenchymal stem cell-educated macrophages into animal models.

The reconstruction of chronic skin wounds remains a public health challenge in dermatology. Precisely controlling and monitoring the wound-healing process should result in enhanced outcomes for the patient. Cell-based therapies have shown great potential in medicine due to their immunomodulatory and healing properties. Herein, we produced activated macrophages by treating circulating monocytes with mesenchymal stem cell (MSC) supernatant. We also demonstrated the critical role of activated macrophages transplantation using amniotic membranes in accelerating wound healing in an animal wound model. The activated macrophages not only exhibited immunomodulatory cytokines like transforming growth factorß (TGFß) and interleukin 10 (and IL10) secretion but also showed attachment and proliferation ability on the amniotic membrane scaffold. Moreover, MSCs supernatant-treated cells also displayed significant ARG1, CD206, and IL 10 genes expression. Inspired by the in vitro results, we examined the in vivo therapeutic efficacy of the activated macrophage transplantation using an acellular amniotic membrane carrier in a full-thickness cutaneous wound model. The wound healing rate was significant in the group treated with macrophages generated via mesenchymal cell therapy seeded human amniotic membrane. There was less scarring in the wound sites after placing cell-scaffold constructs in the wound sites in the animal models. Overall, macrophages stimulated with mesenchymal cells' supernatant exhibited improved healing processes in incisional wounds by decreasing the inflammatory phase, increasing angiogenesis, and reducing scar tissue development.

Alum and metoclopramide synergistically enhance cellular and humoral immunity after immunization with heat-killed Salmonella typhimurium vaccine.

Typically, the killed form of microorganisms in combination with alum does not produce strong cellular immune responses. A recent investigation has indicated the role of dopamine D2 receptor antagonists like metoclopramide in reducing the polarization of immune responses toward Th2 immunity. This study was performed to evaluate the effects of a combination of alum and metoclopramide on the induction of cellular and humoral immunity in response to a heat-killed preparation ofSalmonella typhimurium(HKST). Wistar rats were immunized with the HKST vaccine alone or in combination with alum, metoclopramide, or the alum-metoclopramide mixture twice with a two-week interval. Fourteen days after the last vaccination, immune responses against S. typhimurium and the protective potential of the vaccines were assessed. The combination of alum and metoclopramide as an adjuvant augmented the potential of the HKST vaccine to enhance lymphocyte proliferation, delayed-type hypersensitivity reaction, and antibody titer. These results were concurrent with the polarization of immune response towards the Th1 response and improving protective immunity against S. typhimurium. Overall, the combination of alum and metoclopramide as an adjuvant synergistically enhanced cellular and humoral immunity after immunization with the HKST vaccine.

Molecular effects of curcumin on the experimental autoimmune encephalomyelitis.

Experimental autoimmune encephalomyelitis (EAE) is an animal model of multiple sclerosis (MS). Previous studies have shown that myelin degradation during MS and EAE resulted in reduced expression of some of the proteins, e.g., the MBP (myelin basic protein), and increased expression of genes such as iNOS (Inducible nitric oxide synthase) and NOGO-A in the affected patients. In the present study, EAE was induced by immunizing Wistar rats (n=12) with homogenized spinal cord of guinea pig and Freund's complete adjuvant. Curcumin is an active ingredient in turmeric with anti-inflammatory properties, which has been studied in this article. In this study, the effect of curcumin administration on the change of the expression of MBP, NOGO-A, and iNOS genes was evaluated using the RT-PCR (Reverse transcription-polymerase chain reaction) technique. The obtained results indicated it could be concluded that curcumin was able to improve EAE by increasing the amount of MBP gene expression and reducing the intensity of NOGO-A expression.

The effects of 17 Beta-Estradiol primed mesenchymal stem cells on the biology of co-cultured neutrophil.

OBJECTIVES: Mesenchymal stem cells (MSCs) can influence immune effector cells. It is proved that MSCs respond to various Toll-like receptor (TLR) ligands, which could ultimately result in changes in their immunomodulatory effects. Neutrophils play an essential role in the first line defense system and their function can be regulated by MSCs. Estrogen is a female hormone that contributes to sex differences in several immune-related diseases. With regard to the stated facts, this research aims to elucidate the effects of estrogen treatment on the ability of TLR4-primed MSCs to regulate neutrophil functions. METHODS: Following isolation and characterization, MSCs were stimulated with LPS as a TLR4 ligand and subsequently incubated with different concentrations (0, 10, 20 and 40 nM) of estrogen for 48 hrs. Then, MSCs were co-cultured with neutrophils to investigate the vitality and function of the co-cultured neutrophils. RESULTS: Our results indicated that TLR4-primed MSCs could decrease the viability and neutral red uptake potential of co-cultured neutrophils. Furthermore, neutrophils co-cultured with TLR4-primed MSCs exhibited a decrease in the respiratory burst intensity after being challenged with opsonized yeast. Interestingly, treating TLR4-primed MSCs with estrogen reversed the observed alterations in neutrophil functions. CONCLUSION: It appears that estrogen can alter the interaction between MSCs and neutrophils.

Preparation of Cu nanoparticles fixed on cellulosic walnut shell material and investigation of its antibacterial, antioxidant and anticancer effects.

In this study, antibacterial, antioxidant, and anticancer effects of Cu nanoparticles (CuNPs) fixed on cellulosic walnut shell material were investigated. Firstly, three types of walnut shell-supported copper nanoparticles with various sizes (CuNP-WS1 15-22 nm, CuNP-WS2 60-80 nm and, CuNP-WS3 aggregated of metallic nanoparticles) were synthesized. Antibacterial properties of CuNPs were studied on three strains of bacteria; Staphylococcus aureus, Escherichia coli, and Listeria monocytogenes. DPPH (1, 1-Diphenyl-2-picrylhydrazyl) method was used to examining antioxidant properties. Cytotoxic effects of the synthesized nanoparticles on the cancer cell line were studied. Antimicrobial properties of CuNPs showed that these nanomaterials affect both Gram-positive and Gram-negative bacteria. The antioxidant properties of CuNPs increased significantly by increasing the concentration to 10%. CuNPs appeared to have a dose-dependent cytotoxic effect on K562 cells. However, the IC50 of the synthesized nanoparticles against the K562 (25.24 ± 5 µg/mL) cancer cells was lower significantly (P < 0.01) of the IC50 of these compounds against PBMCs (42.54 ± 6.2 µg/mL).

Immunomodulatory benefits of mesenchymal stem cells treated with Caffeine in adjuvant-induced arthritis.

PURPOSE: We intend to assess the effect of the conditioned medium of Caffeine pulsed MSCS in the amelioration of rheumatoid arthritis (RA)-afflicted rats. METHODS: MSCs were incubated with 0, 0.1, 0.5 or 1 mM Caffeine for 2 weeks. RA was induced by the injection of complete Freund's adjuvant (CFA) into the base of the tail of Wistar rats. According to in vitro studies, RA rats were intraperitoneally treated with MSCs, Caffeine (0.5 mM) pulsed MSCs or vehicle on day 14 when all rats had shown signs of RA. RESULTS: Our results suggest that the least effective dose concentration of Caffeine that can induce potent anti-inflammatory property in the MSC population is 0.5 mM. Without any significant impact on the vitality or MScs' marker, Caffeine at this concentration could induce lower levels of IFN-γ, IL-6, and IL-1ß and a higher level of IDO, TGF-ß, and IL-10 compared to other groups. Therefore, MSCs pulsed with Caffeine at 0.5 mM concentration was selected for in vitro studies. Caffeine pulsed MSCs could reduce the severity of the disease and improve weight-gaining more profoundly than treatment with MSCs alone. Furthermore, Caffeine pulsed MSCs caused a significant reduction in the serum levels C-reactive protein, Nitric oxide, Myeloperoxidase, TNF-α and conversely led a significant increase in the levels of IL-10 more prominent than the similar findings brought about by MSCs alone. CONCLUSION: In general, caffeine-treated MSCs may be a promising strategy for cell-based therapy of RA.

Synergistic benefits of Nicotine and Thymol in alleviating experimental rheumatoid arthritis.

PURPOSE: Given to the anti-inflammatory effect of Nicotine and Thymol, this study was done to evaluate the effects of co-administration of Nicotine and Thymol on the clinical aspects, and immunity responses in Freund's complete adjuvant (FCA)-induced RA in Wistar rat. METHODS: The study population contained a total of 50 male Wistar rats with a weight range 150 ±â€¯7 g, which RA was induced through FCA at them. These animals were randomly allocated into five groups (n = 10): RA rats treated with PBS (100 mg/kg orally), RA rats treated with Thymol (100 mg/kg orally), RA rats treated with Nicotine (2.5mg/kg-orally), and RA rats treated with combined Nicotine and Thymol (half doses with each one-orally). All treatments were initiated at day seven p.i. when all rats showed a clinical score of ≥1. Clinical symptoms of the disease were recorded every other day until the day 23 p.i. RESULTS: Obtained data revealed the combination therapy reduced the severity of the disease and improved weight-gaining more profound than each medication alone. Furthermore, combination therapy caused a reduction in some hematological and biochemical RA parameters, such as Rheumatoid factor, C-Reactive Protein, Nitric oxide, Myeloperoxidase, IL-1, and IL-17 more impressive than each treatment alone. Interestingly, the combination therapy with half doses of Nicotine and Thymol did not have any synergistic advantage in anti-proliferation effect, and therefore immunosuppression side effect compared with using each of agents alone. CONCLUSION: Collectively, it is possible that combination therapy can be applied as a beneficial strategy to control RA.

Thymol has beneficial effects on the experimental model of ulcerative colitis.

OBJECTIVE: Thymol, a natural aromatic monoterpene phenol derived from thymus, possesses anti-inflammatory benefits. Here, we evaluated the potential of thymol therapy in improving an animal model of ulcerative colitis. MATERIALS AND METHODS: Luminal instillation of acetic acid was used to induce colitis in male Wistar rats. Treatment groups daily received prednisolone (2 mg/kg, orally) or thymol (100 mg/kg, orally) for 10 consecutive days. Then, the rats were euthanized and tissue specimens were collected for evaluation of cyclooxygenase-2 (COX-2) expression by immunohistochemistry. Furthermore, the levels of total protein, nitric oxide, myeloperoxidase, malondialdehyde, IL-1, IL-6, and TNF-α were monitored in colonic homogenates. Eventually, the relative mRNA expression of IκBα and NF-κBp65 was investigated using reverse-transcriptase PCR (RT-PCR) in colonic homogenates. RESULTS: Both medications could reduce the mortality rate and the clinical scores of ulcerative colitis. The COX-2 expression was significantly reduced in the colons of thymol-treated animals compared to the prednisolone group. Also, the myeloperoxidase activity, nitric oxide level and malondialdehyde intensity were decreased in the colons of thymol-treated animals to a greater extent compared to the prednisolone group. Moreover, the total protein content of guts showed significant increases in the guts of thymol-treated animals in comparison to the prednisolone group. Nonetheless, thymol significantly reduced the levels of IL-6, and IL-1 compared to prednisolone. Both medications caused a significant decrease in the mRNA level of NF-κBp65, though the mRNA level of IκBα did not show significant changes between the groups. CONCLUSION: Thymol may be a promising agent to ameliorate ulcerative colitis.

The effects of transplanted mesenchymal stem cells treated with 17-b estradiol on experimental autoimmune encephalomyelitis.

The present study was conducted aimed at exploring the modulatory effects of 17-b estradiol (17-bED) on mesenchymal stem cells (MSCs) in the EAE (experimental autoimmune encephalomyelitis) animal model of multiple sclerosis (MS). Following the isolation of bone marrow-derived MSCs from the bilateral femurs and tibias of the male Wistar rats, the cells were harvested and cultured in the presence of 100 nM 17-bED for 24 h. EAE was induced in male Wistar rats (8-12 weeks old) using guinea pig spinal cord homogenate, in combination with the complete Freund's adjuvant. The MSC therapy was triggered when all of the animals obtained a disability score. The symptoms were monitored on a daily basis throughout the study until the rats were euthanized. The mRNA expression of cytokines, including IL-17, IFN-γ, TNF-α, IL-10, IL-4, and TGF-ß together with MMP8 and MMP9 as the family members of matrix metalloproteinases (MMPs) in the brain and spinal cord tissues were examined using real-time PCR. The levels of splenocytes-originated IL-10 and IFN-γ cytokines were also measured by ELISA. The MTT-based research findings showed that the infiltration of lymphocytes into the spleen decreased considerably. It was also observed that the mRNA expression of proinflammatory cytokines decreased significantly, while the mRNA levels of anti-inflammatory cytokines increased remarkably. It was also found that the mRNA levels of the examined matrix metalloproteinases (MMP8 and MMP9) were downregulated significantly. The findings of the present study indicated that the administration of 17-bED enhanced the efficacy of MSCs transplantation and modulated immune responses relatively in the EAE model, via the regulation of either pro- or anti-inflammatory cytokines and matrix metalloproteinases.

Beneficial Effects of Hydroalcoholic Extract of Saffron in Alleviating Experimental Autoimmune Diabetes in C57bl/6 Mice.

Streptozocin (STZ) is a strong alkalizing agent which is capable of destroying the beta cells of the pancreatic islets. Multiple low doses (40 mg/kg, intraperitoneally for 5 consecutive days) prescription of STZ to mice can lead to the T cell-dependent immune response and induction of autoimmune diabetes (AD) with complete similarity to the human type 1 diabetes (T1D). This study has evaluated the effects of hydroalcoholic extract of saffron on the clinical and immunological profile of experimental autoimmune diabetes in C57BL/6 mice. After the establishment of the AD, mice were treated orally with hydroalcoholic extract of saffron (500 mg/kg) for 3 weeks. The results with p<0.05 were considered significant. Obtained data showed that treatment with the hydroalcoholic extract of saffron significantly reduced the incidence of hypoglycemia and restored insulin secretion and histopathological changes in pancreas sections. In addition, treatment with saffron reduced lymphocyte proliferation index in the cells isolated from the pancreas of diabetic mice. Also, the extract of saffron markedly decreased the production of pro-inflammatory interleukin-17 (IL-17) increased anti-inflammatory IL-10 and transforming growth factor-ß in the pancreatic cell population. Moreover, the production of proinflammatory nitric oxide and reactive oxygen substances were down-regulated by the saffron extract. It seems that the hydroalcoholic extract of saffron can be considered as a useful strategy in the treatment of type 1 diabetes.

Comparison of the effects of 17ß- estradiol treated and untreated mesenchymal stem cells on ameliorating animal model of multiple sclerosis.

OBJECTIVES: The current investigation was undertaken to evaluate the effects of 17ß- estradiol (17ß-ED) on the potential of the mesenchymal stem cells (MSCs) for modulation of immunity responses in an animal model of multiple sclerosis (MS). MATERIALS AND METHODS: After isolation of MSCs, cells were cultured in presence of 100 nM 17ß-ED for 24 hr. Modeling of experimental autoimmune encephalomyelitis (EAE) was achieved by using guinea pig spinal cord homogenate, in addition to complete Freund's adjuvant in male Wistar rats. The processes of cell therapy were started following 12 days post-immunization. This duration allows all animals to develop a disability score. The achieved EAE clinical symptoms were regularly monitored every day until day 36, when all of examined rats were euthanized. RESULTS: Cell therapy in the EAE rats with 17ß-ED-primed MSCs exhibited more desirable consequences, which in turn lead to regression of the cumulative clinical score and neuropathological changes that are more than the therapy with untreated MSCs. The serum measures of myeloperoxidase (MPO), nitric oxide (NO) as well as splenocytes-originated pro-inflammatory interleukin-17 (IL-17) and tumor necrosis factor alpha (TNF-α) were significantly decreased in EAE rats treated by 17ß-ED primed-MSCs compared to EAE rats that received untreated MScs. CONCLUSION: Combination of 17ß-ED and MSCs more effectively improved the signs and symptoms of EAE.

Nicotine Augments the Beneficial Effects of Mesenchymal Stem Cell-based Therapy in Rat Model of Multiple Sclerosis.

Experimental autoimmune encephalomyelitis (EAE) in rats through immunization with guinea pig spinal cord homogenate (GPSCH) produces a chronic disease with a relapsing pattern such as multiple sclerosis (MS) in humans. In previous studies, the immunomodulatory benefits of mesenchymal stem cells (MSCs) and nicotine have already been determined. Thus, this research was conducted to assess the additional benefits of the combination therapy of MSCs and nicotine in a rat model of MS. EAE was induced by GPSCH and complete Freund's adjuvant (CFA) in female Wistar rats. The therapies were initiated at day 12 post-immunization (p.i.), when the rats developed a neurological disability score. The symptoms were recorded daily until day 33, when the rats were sacrificed. Finally, the splenocytes were evaluated by Enzyme-linked immunosorbent assay (ELISA) for cytokine production. The therapeutic treatment in the EAE rats with a combination of MSCs and nicotine exhibited a more desirable outcome, causing the regression of the average mean clinical score and neuropathological features to be more favorable than the treatment with either therapy alone. The combination therapy led to a significant reduction in the cumulative disease disability from day 21. For the EAE rats treated with nicotine and MSCs, this period was started from day 22 and 28 p.i., respectively. Besides the increase in the levels of IL-10, the combined therapy significantly reduced the splenocytes production of pro-inflammatory IL-17 as well as TNF-α more profoundly than either of the medications alone. In conclusion, the combination of MSCs and nicotine can be suggested as a promising strategy for further MS therapeutics improvement.

Caffeine Augments The Instruction of Anti-Inflammatory Macrophages by The Conditioned Medium of Mesenchymal Stem Cells.

OBJECTIVES: Mesenchymal stem cells (MSCs) have been shown to produce adenosine, express adenosine receptors, and communicate with macrophages and other cells. However, there is no information about the role of caffeine, as a popular drink and adenosine antagonist, on the crosstalk between MSCs and immune cells. The aim of the current study is to evaluate the effects of the conditioned medium of MSCs treated with caffeine on macrophages. MATERIALS AND METHODS: In this experimental study, MSCs were isolated from bone marrow of rats and pulsed with different concentrations of caffeine (0, 0.1, 0.5 and 1 mM) for 72 hours. The conditioned medium of MSCs was collected after 24 hours, then incubated with macrophages for 24 hours. Finally, the functions of the macrophages were evaluated. RESULTS: Conditioned medium of MSCs treated with caffeine significantly enhanced phagocytosis and simultaneously regressed expression of reactive oxygen species (ROS) and nitric oxide (NO) as well as IL-12 by macrophages compared to the supernatants of MSCs alone. The conditioned medium of MSCs pulsed with caffeine at low to moderate concentrations preserved the neutral red uptake by macrophages and elevated IL-10 secretion by macrophages. A high concentration of caffeine could interfere with the two latter effects of supernatants of MSCs on the macrophages. CONCLUSIONS: Collectively, caffeine treatment of MSCs appeared to augment the instruction of anti-inflammatory macrophages by conditioned medium of MSCs. These findings might offer new insight into the potential mechanisms that underlie the immunomodulatory and anti-inflammatory effects of caffeine.

Nicotine can modulate the effects of the mesenchymal stem cells on neutrophils.

PURPOSE: It has been revealed that mesenchymal stem cells (MSCs) express some of the nicotinic receptor subunits. Moreover, the crosstalk between MSCs and neutrophils is not far-fetched. Therefore, the aim of the present study is to determine the role of nicotine on the effects of MSCs on neutrophils. METHODS: After the isolation of mesenchymal stem cells from the bone marrow of rats, these cells were pulsed with different concentrations of nicotine (0, 0.1, 0.5, and 1µM) for different periods (24, 48, and 72h). Then, the neutrophils were co-cultured with MSCs for 4h and the functions of neutrophils were evaluated. RESULTS: The obtained findings showed that MSCs pulsed with nicotine significantly enhanced the viability and the phagocytic activity of co-cultured neutrophils and simultaneously, decreased the production of reactive oxygen substances (ROS), induced by f-MLP in neutrophils, more profound than MSCs without treatment. Moreover, MSCs, pulsed with nicotine at low to moderate concentrations, preserved the respiratory burst, triggered by opsonized yeast in neutrophils. Nevertheless, a high concentration of nicotine can interfere with the latter aspect of the crosstalk between MSCs and neutrophils. CONCLUSION: The obtained data can offer a new insight into the potential mechanisms, underlying the immunomodulatory effects of nicotine.

Comparison between in-house indirect ELISA and Dot-ELISA for the diagnosis of Fasciola gigantica in cattle.

This survey was done to investigate the efficacy of the in-house indirect ELISA (iELISA) and Dot-ELISA methods Prepared from excretion-secretory (ES Ag) and Crude (Cr Ag) antigens of Fasciola for sero-diagnosis of Fasciola gigantica in cattle. The liver specimens of slaughtered cattle were collected and their liver examined macroscopically and microscopically for infestation to fasciolosis. Sera from two groups of cattle, one infected with fasciolosis (n = 60) and the other non-infected with fasciolosis (n = 60), were used in the iELISA and Dot-ELISA test; grouping based on histopathology results. Except specificity, other parameters such as, sensitivity, accuracy, positive and negative predictive values of both Dot-ELISA and iELISA done with ES Ag were better than those of tests performed with Cr Ag. Interestingly, the reliability of two methods was very good similar to one another.

Estragole and methyl-eugenol-free extract of Artemisia dracunculus possesses immunomodulatory effects.

OBJECTIVE: Some evidence suggests that chronic uptake of estragole and methyl-eugenol, found in the essential oil of Artemisia dracunculus (tarragon), may be associated with an increased risk of hepato-carcinogenicity. The present study was conducted to investigate the immumodulatory and anti-inflammatory potentials of estragole and methyl-eugenol free extract of tarragon. MATERIALS AND METHODS: Aqueous, hydroalcoholic, methanol and hexane extracts of dried and milled tarragon was prepared and analyzed by GC-MS. The estragole and methyl-eugenol free extract was characterized and used for evaluation of immunity in NMRI mice after challenging with sheep red blood cells. RESULTS: It was shown that the aqueous extract of tarragon was free from potentially harmful estragole or methyl-eugenol. Moreover, the immunomodulatory effect of the aqueous extract of tarragon (100 mg/kg for 21 consecutive days) was investigated. The extract significantly increased the level of anti-sheep red blood cells (SRBC (antibody and simultaneously decreased the level of cellular immunity in the treatment group. Moreover, tarragon caused a significant reduction in the production of pro-inflammatory IL-17 and IFN-γ in parallel with a reduction in the ratio of INF-γ to Il-10 or IL-17 to IL-10 in the splenocytes. In addition, the levels of the respiratory burst and nitric oxide production in peritoneal macrophages were significantly decreased. Additionally, the phagocytosis potential of macrophages was significantly increased in treated mice. CONCLUSION: These data showed that the aqueous extract of tarragon may be used as a natural source to modulate the immune system, because it can inhibit pro-inflammatory cytokines and induce anti-inflammatory macrophages.