Вплив гемічної гіпоксії на динаміку концентрацій ГФКБ у структурах мозку та сироватці крові щурів

This article clarifies the questions on study of hypoxic influence on distribution of filament and soluble forms of GFAP in various structures of the brain (neocortex, cerebellum, hippocampus, striatum, middle brain, pons) and blood of the rats. Quantitative analysis of the contents of GFAP in the brain structures of hypoxic rats has established that hemic hypoxia results in changes in intracellular levels of GFAP forms and also in updating their ratio, which allows one to assume not only a change in astroglial cells, but also testifies to reorganization in the system of intermediate filaments of astrocytes. The level of GFAP substantially changed in all cerebral formations, which was already investigated in the early terms of hypoxic period. Observations showed that hemic hypoxia exerted a varied influence on expression of neurospecific protein in the different structures of cerebrum of rats. Differences in expression of GFAP can be caused by the regional differences in astroglial cellular population, and also their internal features that define the possible answers to hypoxic damage in different functional and morphological structures of the brain. An increase in expression of the investigated form of protein can explain strengthening of astroglial reactivity, a feature of the brain that appears in various types of pathologies of the CNS. Reactive asters in such exhibit hypertrophy and are characterized by an increased level of GFAP, which is an early and reliable indicator of astroglial pathology. An increase in expression of the investigated form of protein may be explained by strengthening of astroglial reactivity, a feature of the brain that appears in various types of pathologies of the CNS. The contents of GFAP in the blood of adult rats, as a result of the hypoxic influence received from it, can indicate a release of GFAP from damaged astrocytes in the blood flow. Досліджено ефекти дії гемічної гіпоксії на розподіл філаментної та розчинної форм ГФКБ у різних відділах та структурах головного мозку щурів (кора головного мозку, мозочок, гіпокамп, смугасте тіло, середній мозок, вароліїв міст) і сироватці крові щурів. На основі кількісного аналізу вмісту ГФКБ у структурах головного мозку гіпоксичних щурів установлено, що гемічна гіпоксія викликає зміни внутрішньоклітинного вмісту двох форм білка, а також модифікацію їх співвідношення. Це дозволяє припустити не тільки виникнення змін кількості астрогліальних клітин, а й перебудову системи проміжних філаментів астроцитів. Рівень ГФКБ істотно змінювався в усіх мозкових утворах, що досліджуються вже в ранні терміни постгіпоксичного періоду. Спостереження показали, що гемічна гіпоксія неоднаково впливає на експресію нейроспецифічного білка в різних структурах головного мозку щурів. Відмінності в експресії ГФКБ можуть бути викликані регіональними відмінностями астроцитарної клітинної популяції, а також їх внутрішніми особливостями, які визначають вірогідні відповіді на гіпоксичне пошкодження в різних функціонально та морфологічно структурах мозку. Підвищення експресії досліджуваних форм білка можна пояснити посиленням астрогліальної реактивності – особливістю мозку, що виявляється за різноманітних видів патологій ЦНС. Реактивні астроцити в таких випадках зазнають гіпертрофії та характеризуються підвищеним рівнем ГФКБ, що є раннім та вірогідним індикатором астрогліозу. Вміст ГФКБ у сироватці крові статевозрілих щурів, які зазнали гіпоксичного впливу, може свідчити про вихід із пошкоджених астроцитів у кров’яне русло. Досліджено ефекти дії гемічної гіпоксії на розподіл філаментної та розчинної форм ГФКБ у різних відділах та структурах головного мозку щурів (кора головного мозку, мозочок, гіпокамп, смугасте тіло, середній мозок, вароліїв міст) і сироватці крові щурів. На основі кількісного аналізу вмісту ГФКБ у структурах головного мозку гіпоксичних щурів установлено, що гемічна гіпоксія викликає зміни внутрішньоклітинного вмісту двох форм білка, а також модифікацію їх співвідношення. Це дозволяє припустити не тільки виникнення змін кількості астрогліальних клітин, а й перебудову системи проміжних філаментів астроцитів. Рівень ГФКБ істотно змінювався в усіх мозкових утворах, що досліджуються вже в ранні терміни постгіпоксичного періоду. Спостереження показали, що гемічна гіпоксія неоднаково впливає на експресію нейроспецифічного білка в різних структурах головного мозку щурів. Відмінності в експресії ГФКБ можуть бути викликані регіональними відмінностями астроцитарної клітинної популяції, а також їх внутрішніми особливостями, які визначають вірогідні відповіді на гіпоксичне пошкодження в різних функціонально та морфологічно структурах мозку. Підвищення експресії досліджуваних форм білка можна пояснити посиленням астрогліальної реактивності – особливістю мозку, що виявляється за різноманітних видів патологій ЦНС. Реактивні астроцити в таких випадках зазнають гіпертрофії та характеризуються підвищеним рівнем ГФКБ, що є раннім та вірогідним індикатором астрогліозу. Вміст ГФКБ у сироватці крові статевозрілих щурів, які зазнали гіпоксичного впливу, може свідчити про вихід із пошкоджених астроцитів у кров’яне русло.

Left-right symmetry in 5D and neutrino mass in TeV scale gravity models

We construct a left-right symmetric model based on the gauge group $SU(2)_L\times SU(2)_R\times U(1)_{B-L}$ in five dimensions where both the gauge bosons and fermions reside in all five dimensions. The orbifold boundary conditions are used not only to break the gauge symmetry down to $SU(2)_L\times U(1)_Y\times U(1)_{Y'}$ but also to ``project'' the right handed neutrino out of the zero mode part of the spectrum, providing a new way to understand the small neutrino masses without adding (singlet) bulk neutrinos. This formulation of the left-right model has also two new features: (i) it avoids most existing phenomenological bounds on the scale of the right handed $W_R$ boson allowing for the possibility that the right handed gauge bosons could have masses under a TeV, and (ii) it predicts a stable lepton with mass of order of the inverse radius of the fifth dimension.Comment: 20 pages; some new materials and references adde

Double-blind, 12 month follow-up, placebo-controlled trial of mifepristone on cognition in alcoholics: the MIFCOG trial protocol

Background: Increased levels of cortisol during acute alcohol withdrawal have been linked to cognitive deficits and depression. Preclinical research found that the glucocorticoid Type II receptor antagonist, mifepristone, prevented some of the neurotoxic effects of withdrawal and memory loss. Clinical trials have shown mifepristone effective in the treatment of depression. This study aims to examine the extent to which the glucocorticoid Type II receptor antagonist, mifepristone, when given to alcohol dependent males during the acute phase of alcohol withdrawal, will protect against the subsequent memory loss and depressive symptoms during abstinence from alcohol. Methods/Design: The study is a Phase 4 therapeutic use, “Proof of Concept” trial. The trial is a double-blind randomised controlled clinical trial of mifepristone versus inactive placebo. The trial aims to recruit 120 participants referred for an inpatient alcohol detoxification from community alcohol teams, who meet the inclusion criteria; 1) Male, 2) Aged 18–60 inclusive, 3) alcohol dependent for 5 or more years. A screening appointment will take place prior to admission to inpatient alcohol treatment units to ensure that the individual is suitable for inclusion in the trial in accordance with the inclusion and exclusion criteria. On admission participants are randomised to receive 600 mg a day of mifepristone (200 mg morning, afternoon and evening) for 7 days and 400 mg for the subsequent 7 days (200 mg morning and evening) or the equivalent number of placebo tablets for 14 days. Participants will remain in the trial for 4 weeks (at least 2 weeks as an inpatient) and will be followed up at 3, 6 and 12 months post randomisation. Primary outcome measures are cognitive function at week 3 and 4 after cessation of drinking and symptoms of depression over the 4 weeks after cession of drinking, measured using the Cambridge Neuropsychological Test Automated battery and Beck Depression Inventory, respectively. Secondary outcome measures are severity of the acute phase of alcohol withdrawal, alcohol craving, symptoms of protracted withdrawal and maintenance of abstinence and levels of relapse drinking at follow-up. Discussion: The current trial will provide evidence concerning the role of glucocorticoid Type II receptor activation in cognitive function and depression during acute alcohol withdrawal and the efficacy of treatment with mifepristone

Decreased hippocampal translocator protein (18 kDa) expression in alcohol dependence: a [11C]PBR28 PET study

Repeated withdrawal from alcohol is clinically associated with progressive cognitive impairment. Microglial activation occurring during pre-clinical models of alcohol withdrawal is associated with learning deficits. We investigated whether there was microglial activation in recently detoxified alcohol-dependent patients (ADP), using [11C]PBR28 positron emission tomography (PET), selective for the 18kDa translocator protein (TSPO) highly expressed in activated microglia and astrocytes. We investigated the relationship between microglial activation and cognitive performance. Twenty healthy control (HC) subjects (45±13; M:F 14:6) and nine ADP (45±6, M:F 9:0) were evaluated. Dynamic PET data were acquired for 90 min following an injection of 331±15 MBq [11C]PBR28. Regional volumes of distribution (VT) for regions of interest (ROIs) identified a priori were estimated using a two-tissue compartmental model with metabolite-corrected arterial plasma input function. ADP had an ~20% lower [11C]PBR28 VT, in the hippocampus (F(1,24) 5.694; P=0.025), but no difference in VT in other ROIs. Hippocampal [11C]PBR28 VT was positively correlated with verbal memory performance in a combined group of HC and ADP (r=0.720, P<0.001), an effect seen in HC alone (r=0.738; P=0.001) but not in ADP. We did not find evidence for increased microglial activation in ADP, as seen pre-clinically. Instead, our findings suggest lower glial density or an altered activation state with lower TSPO expression. The correlation between verbal memory and [11C]PBR28 VT, raises the possibility that abnormalities of glial function may contribute to cognitive impairment in ADP

Kinin B1 Receptor Enhances the Oxidative Stress in a Rat Model of Insulin Resistance: Outcome in Hypertension, Allodynia and Metabolic Complications

BACKGROUND: Kinin B(1) receptor (B(1)R) is induced by the oxidative stress in models of diabetes mellitus. This study aims at determining whether B(1)R activation could perpetuate the oxidative stress which leads to diabetic complications. METHODS AND FINDINGS: Young Sprague-Dawley rats were fed with 10% D-Glucose or tap water (controls) for 8-12 weeks. A selective B(1)R antagonist (SSR240612) was administered acutely (3-30 mg/kg) or daily for a period of 7 days (10 mg/kg) and the impact was measured on systolic blood pressure, allodynia, protein and/or mRNA B(1)R expression, aortic superoxide anion (O(2)(*-)) production and expression of superoxide dismutase (MnSOD) and catalase. SSR240612 reduced dose-dependently (3-30 mg/kg) high blood pressure in 12-week glucose-fed rats, but had no effect in controls. Eight-week glucose-fed rats exhibited insulin resistance (HOMA index), hypertension, tactile and cold allodynia and significant increases of plasma levels of glucose and insulin. This was associated with higher aortic levels of O(2)(*-), NADPH oxidase activity, MnSOD and catalase expression. All these abnormalities including B(1)R overexpression (spinal cord, aorta, liver and gastrocnemius muscle) were normalized by the prolonged treatment with SSR240612. The production of O(2)(*-) in the aorta of glucose-fed rats was also measured in the presence and absence of inhibitors (10-100 microM) of NADPH oxidase (apocynin), xanthine oxidase (allopurinol) or nitric oxide synthase (L-NAME) with and without Sar[D-Phe(8)]des-Arg(9)-BK (20 microM; B(1)R agonist). Data show that the greater aortic O(2)(*-) production induced by the B(1)R agonist was blocked only by apocynin. CONCLUSIONS: Activation of kinin B(1)R increased O(2)(*-) through the activation of NADPH oxidase in the vasculature. Prolonged blockade of B(1)R restored cardiovascular, sensory and metabolic abnormalities by reducing oxidative stress and B(1)R gene expression in this model

Functional Role of the Polymorphic 647 T/C Variant of ENT1 (SLC29A1) and Its Association with Alcohol Withdrawal Seizures

Adenosine is involved in several neurological and behavioral disorders including alcoholism. In cultured cell and animal studies, type 1 equilibrative nucleoside transporter (ENT1, slc29a1), which regulates adenosine levels, is known to regulate ethanol sensitivity and preference. Interestingly, in humans, the ENT1 (SLC29A1) gene contains a non-synonymous single nucleotide polymorphism (647 T/C; rs45573936) that might be involved in the functional change of ENT1. Our functional analysis showed that prolonged ethanol exposure increased adenosine uptake activity of mutant cells (ENT1-216Thr) compared to wild-type (ENT1-216Ile) transfected cells, which might result in reduced extracellular adenosine levels. We found that mice lacking ENT1 displayed increased propensity to ethanol withdrawal seizures compared to wild-type littermates. We further investigated a possible association of the 647C variant with alcoholism and the history of alcohol withdrawal seizures in subjects of European ancestry recruited from two independent sites. Analyses of the combined data set showed an association of the 647C variant and alcohol dependence with withdrawal seizures at the nominally significant level. Together with the functional data, our findings suggest a potential contribution of a genetic variant of ENT1 to the development of alcoholism with increased risk of alcohol withdrawal-induced seizures in humans

Effects of alcohol preload on attentional bias towards cocaine-related cues

Background Drug and alcohol users have an ‘attentional bias’ for substance-related cues, which is likely to reflect the incentive-motivational properties of those cues. Furthermore, administration of an alcohol preload increases attentional bias for alcohol and tobacco-related cues in heavy drinkers and tobacco smokers, respectively. The present study investigated attentional bias for cocaine cues in cocaine users and non-users following administration of either alcohol or placebo. Method Thirty-two regular cocaine users and 40 non-users took part. Participants were administered alcohol or placebo, and administration was double blind. After drink administration, a Visual Probe task and Modified Stroop task were used to assess attentional bias. Subjective craving and alcohol outcome expectancies were also measured. Results There was a significant interaction between group and drink type on the visual probe task indicating that cocaine users who had received alcohol had increased attentional bias for cocaine pictures compared to non-users and cocaine users who received placebo. The cocaine Stroop revealed no differences between cocaine users and non-users, and no effects of alcohol in either group. Conclusions Alcohol preload in regular cocaine users increases attentional bias for cocaine cues. However, cocaine users who received placebo did not show attentional bias for cocaine stimuli. Future research should investigate the effects of alcohol preload on attentional bias in cocaine-dependent individuals

Evaluating the relationship between amyloid-β and α-synuclein phosphorylated at Ser129 in dementia with Lewy bodies and Parkinson’s disease

INTRODUCTION: Lewy body and Alzheimer-type pathologies often co-exist. Several studies suggest a synergistic relationship between amyloid-β (Aβ) and α-synuclein (α-syn) accumulation. We have explored the relationship between Aβ accumulation and the phosphorylation of α-syn at serine-129 (pSer129 α-syn), in post-mortem human brain tissue and in SH-SY5Y neuroblastoma cells transfected to overexpress human α-syn. METHODS: We measured levels of Aβ40, Aβ42, α-syn and pSer129 α-syn by sandwich enzyme-linked immunosorbent assay, in soluble and insoluble fractions of midfrontal, cingulate and parahippocampal cortex and thalamus, from cases of Parkinson’s disease (PD) with (PDD; n = 12) and without dementia (PDND; n = 23), dementia with Lewy bodies (DLB; n = 10) and age-matched controls (n = 17). We also examined the relationship of these measurements to cognitive decline, as measured by time-to-dementia and the mini-mental state examination (MMSE) score in the PD patients, and to Braak tangle stage. RESULTS: In most brain regions, the concentration of insoluble pSer129 α-syn correlated positively, and soluble pSer129 α-syn negatively, with the levels of soluble and insoluble Aβ. Insoluble pSer129 α-syn also correlated positively with Braak stage. In most regions, the levels of insoluble and soluble Aβ and the proportion of insoluble α-syn that was phosphorylated at Ser129 were significantly higher in the PD and DLB groups than the controls, and higher in the PDD and DLB groups than the PDND brains. In PD, the MMSE score correlated negatively with the level of insoluble pSer129 α-syn. Exposure of SH-SY5Y cells to aggregated Aβ42 significantly increased the proportion of α-syn that was phosphorylated at Ser129 (aggregated Aβ40 exposure had a smaller, non-significant effect). CONCLUSIONS: Together, these data show that the concentration of pSer129 α-syn in brain tissue homogenates is directly related to the level of Aβ and Braak tangle stage, and predicts cognitive status in Lewy body diseases. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13195-014-0077-y) contains supplementary material, which is available to authorized users