Antidepressant Effect and Recognition Memory Improvement of Two Novel Plant Extract Combinations - Antistress I and Anti-stress II on Rats Subjected to a Model of Mild Chronic Stress.

BACKGROUND: Chronic stress is one of the main factors which lead to depression - a psychiatric disorder affecting millions of people and predicted to be the second ranked cause of premature death in 2020. Depression is often associated with cognitive disturbances and memory deficit. Plant based therapy could be effective in the treatment of mild to moderate depression due to its low level of adverse reaction, its good tolerability and compliance. MATERIALS AND METHODS: 72 male Wistar rats, divided in 9 groups were given orally for 8 weeks two combinations of dry plant extracts - Antistress I and Antistress II and five individual dry extracts obtained from Serratula coronata, Hypericum perforatum, Valeriana officinalis, Crataegus monogyna and Melissa officinalis. The animals were exposed to a chronic unpredictable mild stress for 8 weeks. The depression-like symptoms were evaluated with Forced swim test while the assessment of the memory deficit was performed with Novel object recognition test. RESULTS: Antistress II demonstrates antidepressant effect while Antistress I doesn't improve the depressive-like symptoms. The individual extracts of Hypericum perforatum and Valeriana officinalis also possess antidepressant properties. Antistress II improves the cognition as well as the individual extracts of Hypericum perforatum, Valeriana officinalis and especially Serratula coronata. Dry extract from Serratula tend to have the best effect regarding the recognition memory. The effect of Antistress I on memory deficit is negligible. CONCLUSIONS: Antistress II possesses antidepressant effect and improves the recognition memory while Antistress I doesn't demonstrate any of the above-described effects.

25 Hydroxyvitamin D and Cytokines in Multiple Sclerosis.

INTRODUCTION: Clinical trials of patients with multiple sclerosis (MS) have produced inconsistent results for the profile of cytokine secretion in serum and cerebrospinal fluid in patients with multiple sclerosis during periods of relapse and remission. Epidemiological and clinical observations data reveal an association of the changes in vitamin D serum concentration with the risk of developing MS. AIM: To evaluate changes in serum concentrations of 25(OH)D, IL17, IFN-gamma, TGFß1, IL4, IL10 in relapse and remission and their correlation with the severity of disability. PATIENTS AND METHODS: Fifty-three persons (30 clinically healthy controls and 23 patients with relapsing-remitting multiple sclerosis) living between 41° and 42° northern latitude were registered during the astronomical winter period (October 2012- May 2013). -Patients were diagnosed according to Mc Donald 2010 criteria. The degree of neurological deficit was assessed by EDSS. Serum concentrations of 25(OH)D (nmol/l) and cytokines (pg/ml) were tested by ELISA - once for controls and twice for patients (during relapse and remission). RESULTS: In the studied population average levels of 25(OH)D were close to insufficiency, most pronounced in patients in relapse, as differences were not statistically significant. A reverse correlation was found between the levels of 25(OH)D and the deficit in relapse and remission. Concentrations of TGFß1 significantly increased in remission compared with exacerbation and controls. Serum level of IL4 was significantly lower in relapse compared with controls. In remission there was a marked tendency of increase compared with exacerbation. During clinical improvement IL17 and IFN-gamma tended to decrease compared to the average levels in relapse. In both periods, the average concentrations of IFN-gamma in patients were significantly lower compared with controls. No statistically significant differences were found comparing cytokine changes with those of 25(OH)D and deficit. CONCLUSION: Persistent cytokine imbalance in patients compared with controls is a marker for Th1-mediated CNS demyelination. Anti-inflammatory TGFß1, IL4 are indicators of immune response intensity. The deficit severity does not depend on changes of the tested cytokines, but correlates with 25(OH)D levels during periods of relapse and remission.

Experimental study on the role of 5-HT2 serotonin receptors in the mechanism of anti-inflammatory and antihyperalgesic action of antidepressant fluoxetine.

INTRODUCTION: Fluoxetine is an antidepressant that has anti-inflammatory and antihyperalgesic effects in experimental models of pain and inflammation. The AIM of the present study was to determine the role of 5-HT2 receptors in the mechanism of anti-inflammatory and antihyperalgesic action of fluoxetine after single and repeated administration of the drug. MATERIALS AND METHODS: 40 male Wistar rats were randomly divided in five groups (n = 8) treated for 14 days with saline (control), diclofenac (positive control), fluoxetine, cyproheptadine (5-HT2 antagonist), and fluoxetine + cyproheptadine, respectively. We used the experimental model of inflammation induced by intraplantar injection of carrageenan and nociceptive test with mechanical pressure on the inflamed hind paw. RESULTS: Single and repeated administration of fluoxetine showed that it had significant anti-inflammatory and antihyperalgesic effects when compared with the control (p < 0.05). Cyproheptadine did not change significantly the anti-inflammatory effect of fluoxetine in the first 4 hours, after a single administration. At 24 hours the combination did not differ statistically when compared with the control. Cyproheptadin did not change significantly the anti-inflammatory effect of fluoxetine after repeated administration. After prolonged treatment the group that received fluoxetine + cyproheptadine showed a statistically significant increase in paw pressure to withdraw the hind paw compared with that treated with fluoxetine alone (p < 0.05). CONCLUSIONS: Fluoxetine has anti-inflammatory and antihyperalgesic effects in the carrageenan model of inflammation. 5-HT2 receptor mediated its anti-inflammatory effect in single dose treated animals. Spinal 5-HT2 receptors are involved in the antihyperalgesic effect of fluoxetine after repeated administration.

25 Hydroxyvitamin D and Cytokine Profile in Patients With Relapsing-Remitting Multiple Sclerosis.

In experimental allergic encephalomyelitis, the severity of the deficiency is associated with the loss of axons, and it is likely that cytotoxic T-cells 8 (CD8 T) play an important role. In relapsing-remitting multiple sclerosis, there is a correlation between the inflammatory activity in the lesion and the transection of axons. To understand the pathological mechanisms, it is important to evaluate the changes in serum concentrations of pro- and anti-inflammatory cytokines during the disease course. A total of 46 patients and 40 healthy individuals participated in an open-label, prospective, case-control study from 2012 to 2014. The serum concentrations of cytokines were measured using enzyme-linked immunosorbent assay (ELISA). An immune imbalance was observed during relapse and remission phases compared to the control group. During relapse, the levels of interferon-gamma (IFN-γ) were significantly higher compared to those in remission (p=0.017). During remission, there was an improvement in the deficiency (p<0.001), and the anti-inflammatory cytokines transforming growth factor-beta (TGF-ß) and interleukin 4 (IL4) increased compared to those in relapse (p=0.006; p=0.009). A correlation was found between the serum concentrations of tumor necrosis factor-alpha (TNF-α) and Expanded Disability Status Scale (EDSS) during relapse (correlation coefficient: 0.301; significance (Sig.) (2-tailed 0.042). During the exacerbation, there was a moderate relationship between interleukin 17 (IL17) and 25-hydroxyvitamin D (25(OH)D) (P (p-value (probability value) = 0.02)). TNF-α, IFN-γ, IL17, and TGF-ß serum levels are criteria for evaluating immune inflammatory activity during relapse and remission periods.

Effect of atorvastatin and rosuvastatin on learning and memory in rats with diazepam-induced amnesia.

UNLABELLED: During the past decade, evidence has emerged that statins have neuroprotective effects. AIM: The aim of this study was to investigate the effects of atorvastatin and rosuvastatin on learning and memory in rats with diazepam-induced amnesia. MATERIAL AND METHODS: Experiments were carried out on 48 white male Wistar rats, divided into 6 groups, each of 8 rats. The experimental animals were treated per os for 14 days with atorvastatin and rosuvastatin in doses of 10 mg/kg and 20 mg/kg body weight, respectively. To induce amnesia diazepam was administered intraperitoneally in a dose of 2.5 mg/kg bw. Cognitive skills of the animals were examined after the induction of amnesia with active avoidance test using autonomic reflex conditioner (shuttle box) and passive avoidance tests (step-through and step down) (Ugo Basile, Italy). The following parameters were assessed: number of conditioned responses (avoidances), number of unconditioned responses (escapes) and number of intertrial crossings in the active avoidance test; latency of reactions was measured in the passive avoidance tests. RESULTS: We found a significant increase of conditioned responses in atorvastatin treated animals (in a dose of 10 mg/kg bw) in active avoidance training. In the animals treated with rosuvastatin in both doses there was a statistically significant increase of unconditioned responses. In the step-through passive avoidance test there was significant improvement of short-term and long-term memory following administration of atorvastatin (10 mg/kg bw). Rosuvastatin (10 mg/kg bw) preserves long-term memory. In the step-down passive avoidance test, atorvastatin (10 mg/kg bw) and rosuvastatin (10 mg/kg bw and 20 mg/kg bw) preserve long-term memory. CONCLUSIONS: Atorvastatin (10 mg/kg bw) and rosuvastatin (10 mg/kg and 20 mg/kg bw) improve cognitive functions in rats with diazepam-induced amnesia and preserve long-term memory.

Effect of testosterone propionate on erythropoiesis after experimental orchiectomy.

INTRODUCTION: Androgen deficiency anemia occurs most frequently in pharmacogenic suppression of androgen synthesis or with advancing age in men. Bilateral orchiectomy is a surgical modality used in the treatment of metastatic prostate carcinoma. It is accompanied by marked decrease in circulating serum levels of androgens. AIM: The aim of the experimental study was to determine the effect of substitution therapy with testosterone propionate (TP) on some haematological parameters of erythropoiesis in male rats after orchiectomy. MATERIAL AND METHODS: Eighty Wistar male rats with mean weight of 252.3 g were used in the study. The animals were allocated into 2 control orchidectomized groups, 2 sham-operated groups and 4 experimental orchidectomized groups. Testosterone propionate was administered intramuscularly, once a week at a dose of 4 mg and 8 mg per kilogram of body weight for 15 days and for 15 weeks. Erythrocyte count was performed and hemoglobin and hematocrit levels were measured. RESULTS: In the chronic experiment there was a significant decrease in red blood cells and hemoglobin, and a tendency of decrease in hematocrit after orchiectomy. The effect of TP on erythropoiesis in orchiectomised rats is dose-dependent. CONCLUSION: TP replacement therapy in doses of 4 mg/kg and 8 mg/kg has a stimulating effect on erythropoiesis only in chronic administration.

Vitamin D immunomodulatory potential in multiple sclerosis.

Multiple sclerosis (MS) is an autoimmune disease of unknown etiology whose treatment is of limited efficiency and therefore has a high social burden. As it has been suggested that myelin destruction model, the clinical manifestation and the potential of therapeutic response in MS are correlated, it is quite justifiable that we study various factors (genetic, hormonal, environmental) that take part in the autoimmune process in order to improve the control over the disrupted immune regulation. Results from epidemiological and clinical studies clearly suggest that changes in vitamin D serum concentrations are correlated with the magnitude of the risk of developing MS, the phases of relapse and remittance and with gender differences in vitamin D metabolism. Experimental and clinical studies also have established that 25-hydroxy vitamin D (25(OH)D) and 1,25-dihydroxy vitamin D (1,25(OH)2D) exert an immunomodulatory effect in the central nervous system and peripheral organs of the immune system. The standard reference range of vitamin D concentration in serum is 50-80 nmol/l--it provides normal calcium metabolism. Issues that are discussed include the vitamin D serum concentration needed to suppress the aberrant immune response in MS patients; a subgroup of MS patients suitable for vitamin D treatment, the vitamin D being applied in optimally effective and safe dosage. MS prevalence rate in Bulgaria has increased two-fold in 17 years but this is a rather short interval to be able to assume that the gene pool of the population changes. Thus further studies on possible interactions between different environmental factors and these factors' role in the disease pathogenesis are justified and necessary.

Antinociceptive effect of clomipramine through interaction with serotonin 5-HT2 and 5-HT3 receptor subtypes.

INTRODUCTION: Tricyclic antidepressants are used in the treatment of various pain syndromes. The antidepressant clomipramine inhibits predominantly the reuptake of serotonin in the central nervous system. The mechanism of its analgesic effect is not fully understood. The AIM of the present study was to find experimentally any dose-effect dependence in the analgesic effect of clomipramine and the involvement of the 5-HT2 and 5-HT3 receptors in the mechanism of this effect. MATERIAL AND METHODS: Fifty male Wistar rats were used in the study allocated to five groups (10 animals each): a saline treated control group, one positive control group treated with metamizole and three experimental groups treated with intraperitoneally administered clomipramine in doses of 5, 10 and 20 mg/kg bw, respectively. To study the role of 5-HT2 and 5-HT3 receptors in this effect we used another five groups (10 animals each): control, positive control and three experimental groups treated with clomipramine only, clomipramine and granisetrone and clomipramine and cyproheptadine, respectively. Three nociceptive tests were used: the hot plate test, analgesimeter and the acetic acid-induced writhing test. To gauge the antinociceptive action we used the increased latency in the hot plate test expressed as maximum possible effect % (%MPE), the increase in paw pressure to withdraw the hind paw in analgesimeter and decrease in the number of spinal cord writhes in the acetic acid test. RESULTS: Clomipramine in a dose of 20 mg/kg bw significantly increased the %MPE in hot plate test and the pressure to withdraw the hind paw in the analgesimeter when compared with the control. In the acetic acid test clomipramine decreased non-significantly the number of writhes compared with the controls. Granisetrone reduced non-significantly the antinociceptive effect of clomipramine in all tests. Cyproheptadine potentiated the analgesic effect of clomipramine in acetic acid test and decreased it significantly in the hot plate test. In analgesimeter cyproheptadine decreased significantly the paw pressure to withdraw the tested hind paw at 1 hour and non-significantly at 2 hours. CONCLUSION: Clomipramine in the dose of 20 mg/kg bw has a pronounced antinociceptive affect towards thermal and mechanical pain stimulation. The 5-HT2 and 5-HT3 receptor subtypes are very likely involved in the mechanism of this effect.

Clinical and laboratory study of pro-inflammatory and antiinflammatory cytokines in women with multiple sclerosis.

UNLABELLED: Multiple sclerosis (MS) is an autoimmune demyelinating disorder of the central nervous system characterised with a complex system of interactions between proinflammatory and anti-inflammatory cytokines in its course. AIM: The aim of the present study was to investigate the serum levels of cytokines TNF-a, IFN-gamma, IL-4 and IL-10 in female patients with MS and healthy individuals, the changes occurring in the relapse and remission phases of the disease and their correlation with the severity of the neurological deficit. PATIENTS AND METHODS: Thirty-five women with relapsing-remitting MS were examined. The patients' age ranged between 18 and 50 years and MS was verified clinically and by magnetic resonance imaging according to the McDonald criteria. Thirteen of the patients were treated with interferon-beta-1b. The serum concentrations of TNF-a, IFN-y, IL-4 and IL-10 were determined twice - in relapse and in remission - using an enzyme-linked immunosorbent assay (ELISA). The control group consisted of 35 age-matched healthy females. RESULTS: The comparison of cytokine serum concentrations during the two phases of the disease showed significant elevation of the TNF-alpha serum levels in the relapse phase and of IL-4 - in the remission phase. The comparison between the patients and the healthy control subjects demonstrated statistically significant lower concentrations of TNF-a in remission patients and higher concentrations of IL-10 in relapse patients. The patients with interferon-beta-lb treatment showed different profile of cytokine secretion from the patients without interferon-beta-1b treatment. Interferon-beta-1b-treated patients showed significantly lower serum levels of TNF-a and IFN-gamma during the relapse phase and higher TNF-a and IL-10 serum levels during the remission phase compared with the untreated patients. CONCLUSIONS: Serum levels of TNF-a and IL-4 objectively reflect the immune response during relapse and remission of the disease. The severity of neurological deficit as estimated with the expanded disability status scale (EDSS) does not depend on the serum levels of TNF-a, IL-10 and IFN-gamma in the two phases of MS.

Adverse drug reactions after 24-month treatment with two-dosage regimens of betaferon in patients with multiple sclerosis.

UNLABELLED: An open, prospective randomized trial in patients with multiple sclerosis was conducted to investigate the adverse drug reactions after a 24-month Betaferon treatment. AIM: To assess adverse drug reactions after treatment with two-dosage regimens of Betaferon. PATIENTS AND METHODS: Fifty-five patients were included in the study. They were divided into two groups according to the dosage regimen Betaferon was administered. In group A, Betaferon 8MIU was given subcutaneously every other day. In group B, the treatment was carried out using the following dosage regimen: week 1-3 times per week, every other day in the morning, subcutaneously Betaferon in doses of 2 MIU, 4 MIU and 8 MIU, respectively; weeks 2 to 4 inclusive--3 times per week, every other day in the morning, subcutaneously Betaferon in a dose of 8 MIU; week 5 to month 24 inclusive--subcutaneously every other day in the morning Betaferon in a dose of 8 MIU. Paracetamol, ibuprofen, pentoxifylline and prednisolone were used to cope the ADR according to their type and duration. RESULTS: There were significant changes in group B compared with group A: reduction of the flu-like symptoms and local reactions at the end of the first month of treatment, together with lower relative percentage of patients treated for ADR. CONCLUSION: The reduced dosage regimen and corrective treatment reduce the adverse drug reactions and improve drug tolerability.

Physiological and clinical characteristics of andropause.

The process of aging in man involves a lot of functional and structural changes in the body organs and systems. In this review we shall characterize the physiological and clinical manifestations of andropause. We'll review the physiological basis of the ageing process, the age-related changes in the testosterone secretion regulation, and the dynamics of androgen action and active testosterone metabolism. We also investigate the multifactorial etiology of age-related physiological changes--the body undergoes changes in its structure, there is a loss of muscle strength and decline in physical functions. Sexual dysfunction, hypogonadism and psychological changes are also commonly observed symptoms in this condition. Changes of similar kind can also be seen in young males with androgen deficiency. The age-related changes in physiological functions can potentially lead to some important consequences such as reduced physical activity, higher risk of developing specific diseases (ischemic heart disease, diabetes, osteoporosis), diminished capacity to recover after acute diseases, but most often it leads to increased fracture predisposition. All these may eventually affect negatively the self-care capacity of patients making them require a long-term professional care, and lead to severe psychological and social isolation and increased mortality and change in quality of life. To limit the age-related physiological decline in serum testosterone levels, we should be able to tackle the still unresolved important clinical issue--can hormone replacement therapy administered to elderly men improve their functional status, prevent diseases from developing, improve quality of life and reduce fracture risk. The data included in the present review will contribute to determining the potential benefits and risks of testosterone replacement therapy.

A clinical study of multiple sclerosis patients treated with betaferon.

UNLABELLED: We conducted an open prospective randomized study of 58 multiple sclerosis patients who received Betaferon in a dose of 8 MIU every other day for 2 years. AIM: To assess the effect of Betaferon treatment depending on the number of relapses in the previous 2 years and the stage of neurological deficit at baseline. INCLUSION CRITERIA: clinically definite and MRI-confirmed multiple sclerosis (MS), relapsing-remitting course, at least one relapse 2 years before the study; age 18-50 years; neurological deficit scoring--between 1.0-5.0 steps. According to initial EDSS stage, two groups were formed: group A (32 patients with EDSS score < 2.5 steps) and group B (26 patients with EDSS score > or = 3.0 steps). According to the number of relapses in the last 2 years patients were assigned to group C (29 persons with 1 relapse) and group D (29 persons with > or = 2 relapses). RESULTS: Betaferon therapy effectively reduced relapse frequency in all groups. The severity of disability was significantly reduced in younger patients and in patients with milder neurological deficit at baseline. Regression analysis showed strong correlation between therapeutic improvement and baseline severity of disability. CONCLUSIONS: The obtained results suggest that Betaferon can be used to treat patients with low score of neurological deficit, in an early stage of the disease.

Cytokines and disability in interferon-ß-1b treated and untreated women with multiple sclerosis.

BACKGROUND AND AIMS: T-helper (Th) cells involved in the pathogenesis of multiple sclerosis (MS) represent a functionally heterogeneous population defined by their cytokine secretion profile. The effects of immunotherapeutic drugs on the cytokine network are still not fully clarified. This study aimed to investigate serum levels of IFN-γ, TNF-α, IL-4, IL-10 in interferon-ß-1b-treated and untreated women with relapsing-remitting MS (RRMS) in comparison with healthy controls and the relationship between cytokine concentrations and the degree of disability. METHODS: The study included 35 women with RRMS and 35 age-matched healthy controls. The patients were divided in two groups: Group A-without disease modifying treatment; Group B-treated with interferon-ß-1b. Degree of disability was assessed by the Expanded Disability Status Scale (EDSS). Serum cytokine concentrations were measured by ELISA during relapse and remission. RESULTS: Group A showed higher IFN-γ in remission (p = 0.0239) than the controls; Group B had lower IFN-γ during relapse (p = 0.0226) than controls. EDSS in relapse correlated with the levels of IL-10 for Group A (p = 0.015) and with the concentration of IFN-γ for Group B (p = 0.039). Nontreated patients showed higher EDSS in relapse compared to the interferon-ß-1b-treated group (p = 0.005). CONCLUSIONS: We found an imbalance in the patients' cytokine profile, which may be seen as supportive of the hypothesis that demyelination in the central nervous system is mediated by Th1 lymphocytes. IFN-γ is probably one of the important indicators for intensity of the immune reaction and shows promise as a potential biomarker for the therapeutic effect of interferon-ß-1b. The role of IL-10 in the autoimmune process needs further investigation.

Female sex hormones and cytokine secretion in women with multiple sclerosis.

UNLABELLED: Data from experimental and clinical research suggest that sex hormones may influence the autoimmune process in multiple sclerosis (MS). Studies on the hormonal profile of patients with MS and its relation to the disease activity provide heterogeneous results. OBJECTIVES: The aim of this study is to investigate the changes in serum levels of estradiol and progesterone and their correlations with the cytokine profile and the degree of disability in women with relapsing-remitting MS (RRMS). METHODS: The serum concentrations of estradiol, progesterone, tumor necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN-gamma), interleukine-4 (IL-4) and interleukine-10 (IL-10) were measured and the degree of disability was determined in 35 women with RRMS, during relapse and remission. Serum levels of hormones were measured by micro-particle enzyme immunoassay and ELISA was used for the cytokines concentrations. The degree of disability was assessed by the Expanded Disability Status Scale and the Scripps Neurological Rating Scale. RESULTS: Sixty per cent of patients had serum concentrations of estradiol and/or progesterone below the lower limit of normal in one or both phases of MS. Hormonal levels increased significantly during remission in these patients. Women with and without hormonal abnormalities differed in terms of cytokine profile during relapse and remission. Significantly higher TNF-alpha in both phases and IFN-gamma in remission was found for the patients with hormonal disturbances compared to these with normal hormonal status. CONCLUSIONS: Our study finds high frequency of hormonal disturbances among female patients with RRMS. Abnormally low concentrations of sex hormones are associated with higher serum levels of TNF-alpha and IFN-gamma, which could suggest suppressive effect of estradiol and progesterone on pro-inflammatory cytokine secretion.