Depression is one of the current dilemmas in both developed and developing societies. Studies show that the severity of psychiatric symptoms is directly related to the degree of inflammation caused by cytokines secreted by the immune system. Hence, evaluating serum cytokine levels in patients with depression can help to understand the pathogenesis of the disease and make the best therapeutic decisions. The present study investigated the levels of inflammatory cytokines, tumor necrosis factor-α (TNF-α), interleukin-1 (IL-1), and interleukin-6 (IL-6) in patients with major depression or bipolar disorder during depressive episodes (BDDE) before and after a 6-month pharmaceutical intervention. Patients referring to 3 clinics were recruited for the study. The diagnosis of major depression or bipolar disorder in a depressive phase was made according to the Diagnostic and Statistical Manual of Mental Disorders -5(DSM-5) criteria. There was a significant difference in depression levels between the pre-intervention and 6-month follow-up in both groups. After 6 months, IL-1 and IL-6 levels in the bipolar disorder group had decreased while TNF-α levels had increased. There was also a significant difference between pre-intervention and follow-up levels of IL-1.  Serum levels of IL-1 and IL-6 decreased significantly in both groups after the 6-month follow-up, and symptom improvement was observed. TNF-α levels, on the other hand, decreased in the major depression group but increased in the bipolar disorder group. Considering that inflammation is a major outcome of depression, treatment strategies to reduce inflammation could be a practical approach to improving psychiatric symptoms.

Emerging evidence indicates that gut microbiota interacts with immune and nervous systems in the host and plays a critical role in the pathogenesis of multiple sclerosis (MS) and many psychiatric disorders such as depression and anxiety. The aim of this study was to explore the influence of gut bacterial depletion from early adolescence on adult immunological and neurobehavioral responses in mice with experimental-autoimmune-encephalomyelitis (EAE). We used an animal model of gut microbiota depletion induced by antibiotics from weaning to adulthood to assess clinical signs, cognitive function and depression-and anxiety-related symptoms in non-EAE and EAE-induced mice. We measured levels of interferon (IFN)-γ, interleukin (IL)-17A and IL-10 in serum, and BDNF, IL-1β and tumor necrosis factor (TNF)-α) in the hippocampus. Antibiotic-treated mice displayed a significant delay in the onset of clinical symptoms of EAE. However, a higher severity of EAE was found between days 19–22 post-immunization in antibiotics-treated mice, while a reduction in the clinical signs of MS was observed at days 24–25 post-immunization. Antibiotic administration decreased IFN-γ and IL-17A levels and increased IL-10 in serum of EAE-induced mice. Antibiotic treatment significantly decreased hippocampal BDNF and enhanced learning and memory impairments in EAE-induced mice. However, no significant changes were found in non-EAE mice. Non-EAE and EAE mice treated with antibiotics exhibited increased anxiety-related behaviors, whereas depression-related symptoms and increased hippocampal TNF-α and IL-1β were only observed in EAE-induced mice treated with antibiotics. This study supports the view that depletion of gut microbiota by antibiotics from weaning profoundly impacts adult immunological and neurobehavioral responses.

Background: The aim of this study was to provide a simple and efficient way of producing IgY antibodies against Escherichia coli and extract that by polyethylene glycol (PEG) precipitation method. Methods: An ti- Escherichia coli antibody was produced in hens by using formalin-killed E.coli and confirmed by ELISA method, applying serum`antibody. The specific IgY was extracted from egg yolk by Polyethylene Glycol (PEG) Precipitation and analysed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Results: The result of ELISA shows that the IgY titre increased from initial immunization and the high titre (≥0.071) persisted after the first immunization. Heavy chain of IgY with ~68 KDa was observed in the Gel electrophoresis pattern. Conclusion: The results of the ELISA indicates the specificity of the immunoglobulin Y to the target antigen and the result of SDS-PAGE represented the appropriate extraction method. More research must be done on the ability of these antibodies to inhibit the growth of E.coli.

Background: Staphylococcus aureus strains exhibiting multiple antibiotic resistances are isolatedfrom most communities and hospital infections. Treatment of patients with these infections hasbeen difficult. The aim of this study was to detect and extract, the egg yolk immunoglobulin Y asa potential source of anti- S. aureus antibody.Methods: Specific IgY was produced by immunizing hens with formalin-killed S. aureus. Thespecificity of serum`s antibody was confirmed by ELISA method. The antibodies were extractedfrom egg yolk by polyethylene glycol (PEG) precipitation. Proteins were analysed by sodiumdodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE).Results: Chicken egg yolk antibodies (IgY) were raised against S. aureus in the serum afterinjections. Up to 104 dilution specific antibodies were determined in serum.Conclusion: The results of the ELISA indicates the specificity of the immunoglobulin Y to thetarget antigen. In order to find a viable alternative to antibiotic treatments, more research must bedone on the ability of these antibodies to inhibit the growth of S. aureus.

Background: E. coli strains can cause a wide variety of intestinal and extra-intestinal diseases.Increasing occurrence of antibiotic resistance would complicate the treatment of such diseases.Objectives: The aim of this study was to detect the egg yolk immunoglobulin Y as a potential source of anti-Escherichia coli.Materials and Methods: This study was conducted by using killed E.coli with formaldehyde as antigen.Chickens were immunized with 500 µL of antigen diluted in equal volume of Freund's complete adjuvant (FCA).Booster injections of increasing concentrations of antigen were given to raise the antibody level in serum.The specificity of antibody was confirmed by ELISA method.Results: Chicken egg yolk antibodies (IgY) were raised against E. coli in the serum after injections.Up to 1.10000 dilution antibodies were determined in serum, which shows high amount of antibodies are produced in immunized chickens.Conclusions: Further studies have to be conducted to evaluate the potency of these antibodies in treatment of variety of diseases.The outcome of this research work would be an alternative to the current antibiotic treatments.

Background: Progesterone as a sex steroid hormone is thought to affect and prevent demyelination, but its role in promoting myelin repair is far less investigated. In this study, remyelinating potential of progesterone in corpus callosum was evaluated on an experimental model of MS. Methods: In this experimental study, adult male C57BL/6 mice were fed with 0.2% (w/w) cuprizone in ground breeder chow ad libitum for 6 weeks. At day zero, after cuprizone removal, mice were divided randomly into two groups: (a) placebo group, which received saline pellet implant, (b) progesterone group, which received progesterone pellet implant. Some mice of the same age were fed with their normal diet to serve as the healthy control group. Two weeks after progesterone administration, Myelin content was assessed by Luxol-fast blue staining. The myelin basic protein (MBP) and proteolipid protein (PLP) expression were assessed using Western blot analysis and the changes in the number of oligodendrocytes and oligodendroglial progenitor cells were assessed by immunohistochemistry (IHC) and flow cytometry. Results: Luxol-fast blue staining revealed enhanced remyelination in the progesterone group when compared with the placebo group. Densitometry measurements of immunoblots demonstrated that MBP and PLP proteins contents were significantly increased in the progesterone group compared with the placebo group. Flow cytometry and IHC analysis showed increases in Olig2 and O4 cells in the progesterone group compared with the placebo group. Conclusion: Overall, our results indicate that progesterone treatment can stimulate myelin production and that it may provide a feasible and practical way for remyelination in diseases such as multiple sclerosis.

UNLABELLED:Alzheimer's disease (AD) is the most prevalent neurodegenerative disorder. It is characterized by formation of amyloid plaques and neurofibrillary tangles in the brain, degeneration of the cholinergic neurons and neural cell death. This study was aimed to investigate the effect of a triazine derivative, C16H12Cl2N3S, on learning in an Alzheimer's rat model. Animals were divided into seven groups; each group contained seven animals.CONTROL GROUP:animals received no surgery and treatment; saline group: animals received normal saline after recovery; sham group: animals received 10% DMSO after recovery; STZ group (Alzheimer's model): animals received streptozotocin (STZ) in four and six days after recovery; T5, T10 and T15 groups: animals were treated with triazine derivative, C16H12Cl2N3S, at doses of 5, 10 and 15 µM, respectively. All drugs were injected intracerebroventricular. The spatial learning and histological assessment were performed in all groups. Animals in STZ group had more deficits in spatial learning than the control group in Morris water maze. C16H12Cl2N3S improved spatial learning significantly compared to STZ group. The CA1 pyramidal layer thicknesses in STZ group were reduced significantly compared to control group. C16H12Cl2N3S increased the CA1 pyramidal layer thickness in T15 group compared to STZ group. Current findings suggest C16H12Cl2N3S may have a protective effect on learning deficit and hippocampal structure in AD.

BACKGROUND:Cytokines are secreted signaling proteins which play essential roles in immune responses during experimental autoimmune encephalomyelitis (EAE), a demyelinating model that mimics many features of multiple sclerosis (MS). Interleukin 6 (IL-6) is a multifunctional cytokine produced by different cells, mediating inflammatory reactions and immune-mediated processes. Several studies have described immunosuppressive potentials of several herbal medicines. MS14 as an Iranian marine herbal medicine has anti-inflammatory and immunomodulatory activities.OBJECTIVES:The present study investigated the immunosuppressive potential of MS14 as an herbal drug as well as the IL-6 level in EAE model. We hope it will be a new approach for neurologic diseases and autoimmune originated diseases therapy.PATIENTS AND METHODS:The present experimental study was a collaboration between Department of Anatomical Sciences of Tabriz University of Medical Sciences and Shefa Neuroscience Research Center of Tehran. We used 30 C57BL/6 mice. The animals were immunized with myelin oligodendrocyte glycoprotein (MOG) to induce EAE and treated with MS14-containing (30%) diets. Subjects were selected by simple random sampling and then they were randomly allocated to two groups. EAE symptoms were assessed using the standard 10-point EAE scoring system from the seventh to the 35th day after immunization. Afterwards, the spleen was removed and its cells were cultured with or without MOG 35-55; then, the IL-6 level was analyzed by ELISA. In addition, histopathological studies were carried out for demyelination lesion evaluation in the spinal cord.RESULTS:MS14 significantly improved clinical symptoms of EAE compared with the control (P < 0.05). It also suppressed proliferative responses of T cells and decreased IL-6 expression (16.93 ± 2.7 vs. 21.4 ± 3.33) (P < 0.05).CONCLUSIONS:Our results strongly suggested that IL-6 as a potential molecule could have a role in neuroimmunology and neuroinflammation, which is in congruent with previous studies. Therefore, it can be a clear target in strategic therapies and support effective properties of phytotherapy in EAE and MS treatment.

Background: Experimental autoimmune encephalomyelitis (EAE) is an animal model of multiple sclerosis, which is a demyelinating and an inflammatory disease of central nervous system. Recent studies have established that some molecules such as Lipocaline2 (LCN2), which expresses during inflammatory conditions, play an important role in EAE pathogenesis and might involve in its treatment process. Recently, it has been proved that MS14, an herbal-marine drug, has anti-inflammatory properties through reduction of TNF-α and IL-1β. Thus, the present study investigated the effects of MS14 on the course of EAE and its relation to LCN2 expression in both protein and gene levels. Methods: EAE was induced in female C57BL/6 mice using Hooke kits. Animals were scored for clinical signs of the disease according to a 10-point EAE scoring system. On 21 st and 35 th days after immunization, mice (n = 4/group) were deeply anesthetized, and the spinal cords were removed. Inflammatory cell infiltration and LCN2 expression in spinal cord were assessed by hematoxylin and eosin staining, immuno-histochemistry, and real-time PCR methods. Results: MS14 significantly ameliorated EAE symptoms and decreased lymphocyte infiltration into the spinal cord ( P<0.05). Our data also revealed that LCN2 expression was significantly downregulated in acute and chronic phases of EAE both at protein and gene levels after MS14 treatment (P<0.05). Conclusion: The results demonstrated that MS14 regulatory effect on EAE is accompanied by LCN2 downregulation after treatment with the herb; however, more studies are required for clarifying the other involved mechanisms. Iran. Biomed. J. 18 (4): 196-202, 2014

Tropisetron, a selective 5-HT3 receptor (5-HT3R) antagonist, has been widely used to counteract chemotherapy-induced emesis. New investigations described the immunomodulatory properties of tropisetron which may not be 5HT3R mediated. In the present study, we assessed the potential effects of tropisetron on an animal model of multiple sclerosis (MS), experimental autoimmune encephalomyelitis (EAE). EAE was induced in C57BL/6 mice by myelin oligodendrocyte glycoprotein peptide (MOG35-55) immunization. Animals were treated with tropisetron (5 mg/kg/day); m-chlorophenylbiguanide (mCPBG), a selective 5-HT3R agonist (10 mg/kg/day); tropisetron (5 mg/kg/day) plus mCPBG (10 mg/kg/day), and granisetron (5 mg/kg/day) intraperitoneally on days 3-35 post-immunization. Treatment with tropisetron and granisetron markedly suppressed the clinical symptoms of EAE (p<0.001) and reduced leukocyte infiltration as well as demyelination in the spinal cord (p<0.05). In addition, in vivo tropisetron, granisetron or tropisetron plus mCPBG therapy greatly reduced in vitro MOG35-55-stimulated proliferation of mononuclear cells from spleens, and MOG35-55-induced IL-2, IL-6 and IL-17 production by splenocytes isolated from EAE-induced mice (p<0.05). Concurrent administration of tropisetron and mCPBG did not significantly alter the histological damage in the spinal cord. mCPBG had no effect on the mentioned parameters. Taken together, these findings indicate that tropisetron has considerable immunoregulatory functions in EAE and may be promising for the treatment of MS or other autoimmune and inflammatory diseases of the CNS. Furthermore, beneficial effects of tropisetron in this setting seem to be both receptor dependent and receptor independent in the early phase of the disease.

INTRODUCTION:Stroke is one of the most important reasons of death. Hence, trials to prevent or lessen the complications originated by stroke are a goal of public health worldwide. The ischemia-reperfusion causes hypoxia, hypoglycemia and incomplete repel of metabolic waste products and leads to accumulation of free radicals triggering neuronal death. The A1 adenosine receptoras an endogenous ligand of adenosine is known to improve cell resistance to destructive agentsby preventing apoptosis. Vitamin C as a cellular antioxidant is also known as an effective factor to reduce damages initiated by free radicals. We studied the protective effects of A1 receptor agonist in combination with vitamin C against ischemia-reperfusion.METHODS:Ischemia was induced by common carotid artery occlusion in bulb-c mice (20-30 gr). Y-Maze was employed to scale the short-term memory and Nissl staining was used to count the cells in hippocampus.RESULTS:We found that concurrent treatment of A1 receptor agonist and vitamin C significantly reduced neuronal death in CA1. The Memory scores were also significantly improved (P < 0.05).DISCUSSION:Our data point to the therapeutic effects of CPA/vitamin C co-administration and highlight the beneficial role of A1 adenosine receptor signaling in the context of stroke.

Introduction: Bone marrow (BM) is a known source of mesenchymal stem cells (MSCs) that are used for cell therapy. This study attempts to identify if the Wharton’s Jelly (WJ) is a suitable substitute for BM as a source for MSCs. Materials and Methods: A population of human WJ and BM stem cells were isolated and incubated with fluorescein conjugated antibodies for five specific MSC markers. Cell populations were subsequently analyzed by flow cytometry. Results: Most of the stem cells isolated from WJ and BM were positive for MSC markers. No significant differences existed between the expression of typical markers of MSCs in WJ isolated cells and that of BM. Conclusion: WJ is a potential replacement for BM as a source of MSCs and can be used for cell therapy in clinical applications.

OBJECTIVE:Multiple sclerosis (MS) is an immune-mediated demyelinating disease of the central nervous system (CNS). Stem cell transplantation is a new therapeutic approach for demyelinating diseases such as MS which may promote remyelination. In this study, we evaluate the remyelinating potential of adipose mesenchymal stem cells (ADSCs) and their effect on neural cell composition in the corpus callosum in an experimental model of MS.MATERIALS AND METHODS:This experimental study used adult male C57BL/6 mice. Cultured ADSCs were confirmed to be CD73(+),CD90(+), CD31(-),CD45(-), and labeled by PKH26. Animals were fed with 0.2% w/w cuprizone added to ground breeder chow ad libitum for six weeks. At day 0 after cuprizone removal, mice were randomly divided into two groups: the ADSCs-transplanted group and the control vehicle group (received medium alone). Some mice of the same age were fed with their normal diet to serve as healthy control group. Homing of ADSCs in demyelinated lesions was examined by fluorescent microscope. At ten days after transplantation, the mice were euthanized and their cells analyzed by luxol fast blue staining (LFB), transmission electron microscopy and flow cytometry. Results were analyzed by one-way analysis of variance (ANOVA).RESULTS:According to fluorescent cell labeling, transplanted ADSCs appeared to survive and exhibited homing specificity. LFB staining and transmission electron microscope evaluation revealed enhanced remyelination in the transplanted group compared to the control vehicle group. Flow cytometry analysis showedan increase in Olig2 and O4 cells and a decrease in GFAP and Iba-1 cells in the transplanted group.CONCLUSION:Our results indicate that ADSCs may provide a feasible, practical way for remyelination in diseases such as MS.

Methadone is known to exert modulatory effects on the immune system. We investigated the potential effects of methadone on infiltration of inflammatory cells into the spinal cord, as well as the proliferative and cytokine responses of T cells in MOG(35-55)-induced experimental autoimmune encephalomyelitis in mice. Methadone significantly suppressed clinical signs of the disease and level of inflammatory cytokines (p<0.05) produced by T cells. Moreover, invasion of inflammatory cells into the spinal cord was significantly decreased by methadone (p<0.05). Our data point to therapeutic effects of methadone and highlight the beneficial role of opioid receptor signaling in the context of autoimmune neuroinflammation.

Spreading depression (SD) is an intrinsic bioelectrical property of the human central nervous system, which plays a key role in neurological disorders. In the present study, we investigated whether experimentally induced repetitive SD caused neuronal death in cortical and subcortical regions of the juvenile rat brain. The animals were anesthetized and the electrodes as well as a cannula were implanted over the brain. Repetitive cortical SD events were induced by KCl injection. The brains were removed after 4 weeks. Repetitive SD enhanced the production of dark neurons, reduced the mean volume of normal neurons, increased the number of apoptotic neurons, and enhanced expression of the NR2B subunit of NMDA receptors as well as the GluR1 subunit of AMPA receptors in various regions of the juvenile rat brain. In addition, induction of repetitive SD enhanced long-term potentiation in CA1 hippocampal area. We observed a correlation between cell injury/neuronal death induced by repetitive SD and changes in glutamate receptor expression. The data indicate that repetitive cortical SD in juvenile rats causes neuronal damage in both cortical and subcortical areas of the brain. This may play an important role in the pathophysiology of SD-related neurological disorders, especially in children.

arkinson’s disease (PD) is one of the most common neurodegenerative diseases characterized by tremor, stiffness and slowness of motion due to dopamine (DA) depletion following dramatic loss of dopaminergic neurons in Substantia nigra. Unfortunately, this debilitating disease has no curative treatment. Neuroprotective agents might be useful via postponing the initiation of neurodegenerative processess and increasing cell resistance to oxidative stress and neuroinflammation reThe Effects of Boswellia Resin Extract on Dopaminergic Cell line, SK-N-SH, against MPP+-Induced Neurotoxicity

Multiple Sclerosis (MS) is an autoimmune inflammatory, demyelinating disease of human central nervous system. Experimental Autoimmune Encephalomyelitis (EAE) is the commonly used animal model of MS. Calorie restriction has been found to reduce inflammation and autoimmune responses and promote neuroprotection. In this study we evaluated the effects of intermittent feeding protocol of the calorie restriction in a mouse model of EAE. Fifty four female mice (C57BL/6) were used in this study. The animals were divided into two dietary groups: ad libitum (AL) (n = 29) with free access to food and water and intermittent feeding (IF) (n = 25) with access to food on alternate days. After 8 weeks, EAE was induced in animals by immunization with MOG antigen (Hooke labs, Lawrence, MA, USA) subcutaneously. AL and IF groups were then further divided into two groups each: AA (ad libitum until the end of study) (n = 16) and AI (subjected to intermittent feeding regimen after immunization day) (n = 13). The IF group was divided into II (continued intermittent feeding regimen until the end of study) (n = 13) and IA (changed to AL regimen after immunization day) (n = 12). All the animals were behaviorally monitored for 35 days after immunization and observed daily for the signs and severity of disease with EAE scoring scale [0-5] and cumulative disease index (CDI) score. Intermittent feeding significantly reduced the incidence of EAE in IF groups (AI 0%, II 18.5%, IA 22.2%, p < 0.05). In addition, intermittent feeding significantly delayed the onset of EAE in AI group (p < 0.05) and also, intermittent feeding significantly reduced the severity of disease in II and IA groups (AA vs. II, p < 0.05 & AA vs. IA p < 0.05) groups. The CDI was also significantly reduced in intermittent feeding fed groups [AI, II and IA compared to AA group (P < 0.05, <0.01, <0.05 respectively)]. Intermittent feeding regimen protocol of the calorie restriction significantly suppressed EAE incidence, induction, and severity. The results of this study suggest possible role of intermittent feeding in the treatment of Multiple Sclerosis patients.