ATP release during cell swelling activates a Ca2+-dependent Cl - Current by autocrine mechanism in mouse hippocampal microglia

Microglia cells, resident immune cells of the brain, survey brain parenchyma by dynamically extending and retracting their processes. Cl- channels, activated in the cellular response to stretch/swelling, take part in several functions deeply connected with microglia physiology, including cell shape changes, proliferation, differentiation and migration. However, the molecular identity and functional properties of these Cl- channels are largely unknown. We investigated the properties of swelling-activated currents in microglial from acute hippocampal slices of Cx3cr1+/GFP mice by whole-cell patch-clamp and imaging techniques. The exposure of cells to a mild hypotonic medium, caused an outward rectifying current, developing in 5-10 minutes and reverting upon stimulus washout. This current, required for microglia ability to extend processes towards a damage signal, was carried mainly by Cl- ions and dependent on intracellular Ca2+. Moreover, it involved swelling-induced ATP release. We identified a purine-dependent mechanism, likely constituting an amplification pathway of current activation: under hypotonic conditions, ATP release triggered the Ca2+-dependent activation of anionic channels by autocrine purine receptors stimulation. Our study on native microglia describes for the first time the functional properties of stretch/swelling-activated currents, representing a key element in microglia ability to monitor the brain parenchyma

Achievement of target blood pressure in patients with arterial hypertension on the background of antihypertensive therapy in real clinical practice

Aim. To evaluate the features of pharmacotherapy in achieving different levels of target blood pressure (BP) in patients with arterial hypertension (AH) with the absence or presence of comorbid diseases in real outpatient practice.Material and methods. At the open multicenter observational study, outpatient physicians filled original patient questionnaires, which reflected the demographic data of patients, the presence of comorbid diseases and conditions prescribed antihypertensive drugs and achieved during treatment with their use levels of systolic (SBP) and diastolic (DBP) blood pressure (BP), body mass index (BMI), creatinine and blood glucose levels, as well as information about smoking. The obtained data were stratified into groups depending by the level of blood pressure achieved in patients during the therapy, as well as depending on the existing comorbid diseases. Estimated rate of prescription of antihypertensive agents, the number of components of therapy, the number assigned to tableted dosage forms (tablets). We also evaluated the frequency assignments of fixed combinations (FC). Results. The study included data from 2073 patients. They were divided into six groups according to the level of BP achieved. The groups were comparable by demographic and anthropometric characteristics, as well as in gender representation. In patients of the first group on the background of therapy were achieved the lowest values of blood pressure — 120­129/<80 mm Hg. art. They were less likely than other groups to detect comorbid diseases, less frequently prescribed thiazide/thiazide­like diuretics (TD), and FC were prescribed in 33,8%. In patients of the second group the blood pressure level was 130­139/<80 mm Hg. art., the duration of hypertension was the smallest, they were most often prescribed angiotensin II receptor blockers (ARBS) and so on, and the share of FC was the maximum among the compared groups — 42,3%. In the third group, the blood pressure level was 120­139/80­89 mm Hg. St. These patients are most often prescribed angiotensin converting enzyme inhibitors (ACEi), but rarely angiotensin receptors blockers (ARB), frequency assignments of FC — 37,8%. The level of blood pressure in patients of the fourth group who did not achieved the target value of SBP (≥140 mm Hg), in the fifth group — the target value of DBP (>90 mm Hg), and in the sixth group — the target values of SBP (≥140/>90 mm Hg. art.). Their share in the total sample was 19,9%, 4,1%, and 41,2%, respectively. Patients from these groups were more likely to have comorbid diseases, they were more often prescribed four or more components of therapy. BP level <130/<80 mm Hg in patients with type 2 diabetes mellitus (DM 2) was achieved in 4,2%, in patients with coronary heart disease (CHD) in 8,3%. In these groups, a high frequency of beta­blockers (BB) was noted. Patients with chronic kidney disease (CKD) had blood pressure levels of 130139/<80 mm Hg was 7,9%. Among patients with stroke/transient ischemic attack (TIA) blood pressure 120­129/<80 mm Hg was achieved in 2%. In the general sample of patients, one component of antihypertensive therapy was prescribed in 5,8%, two — in 48,3%, three — in 34,7%, four or more — in 11,2%.Conclusion. Target blood pressure <140/90 mm Hg was achieved at 34,8%, and the level of blood pressure <130/80 mm Hg — only at 11,5% of patients. In these patients, comorbid diseases were less often observed, from hypotensive drugs, ACEI, BB or TD were most often used, the predominant appointment of twocomponent antihypertensive therapy was noted, which was most often presented in the form of two tablets. In patients with comorbid diseases revealed a very low proportion of achieving the target level of blood pressure: with DM 2 — 4,2%, with CKD — 7,9%, with IHD — 8,3%, with stroke/TIA — 2%. Among the patients of the whole sample, two­ and three­component antihypertensive therapy was most often prescribed (48,3% and 34,7%, respectively). A greater number of antihypertensive components were prescribed to patients with several comorbid diseases, and, consequently, with a more severe course of hypertension

Farmakoekonomicheskie aspekty umen'sheniya mikroa11'buminurii u bol'nykh gipertonicheskoy bolezn'yu i sakharnym diabetom tipa 2

У больных гипертонической болезнью и СД с МА наиболее оправданным с фармакоэкономических позиций является применение индапамида-ретард и эналаприла в дозе, не превышающей 12 мг/сут. Комбинация кандезартана и лизиноприла экономически и клинически более выгодна, чем монотерапия кандезартаном. Сравнение фармакоэкономической предпочтительности применения различных антигипертензивных препаратов в России и США обнаружило некоторые отличия, что в значительной степени обусловлено исходно разным соотношением цен на оригинальные препараты. Представленные данные выявили различия в фармакоэкономической эффективности оригинальных гипотензивных препаратов, что необходимо учитывать при проведении массовых программ по профилактике и лечению артериальной гипертензии, а также при разработке лекарственных формуляров различного уровня

Barriers to effective outpatient hypertension treatment: a view of physicians and patients

Aim. To study the opinion of primary care physicians and hypertensive (HTN) outpatients with different compliance rate on factors preventing effective antihypertensive therapy (AHT).Material and methods. Primary care physicians and HTN outpatients were questioned. Questionnaires for physicians and patients included informative and survey parts, with a list of possible factors aggravated adherence to treatment. The assessment was carried out using visual analogue scale. The patient questionnaire also included questions about AHT and the eight-item Morisky Medication Adherence Scale (MMAS-8). The calculation was carried out with a 95% confidence interval (CI).Results. The survey involved 298 physicians and 517 patients. Among patients, about 1% had a high compliance rate, 34% — moderate, 65% — low. In all groups, AHT did not significantly differ and was characterized by a low frequency of prescribing fixeddose combinations (27,1%). According to physicians, the most significant and equivalent are the economic aspects of treatment — 7,9±2,1 (95% CI: 7,51-8,38), the need for lifestyle change — 7,9±2,4 (95% CI: 7,37-8,38) and, to a slightly lesser extent, psychological aspects 6,8±2,2 (95% CI: 5,43-6,43). The economic aspects of treatment and need for lifestyle change were also most significant factors according to patients with high (8,8±1,8 (95% CI: 7,23-10,37) and 8,4±1,7 (95% CI: 6,93-9,87), respectively) and low (95% CI: 6,4±3,0 (5,65-7,07) and 6,2±2,8 (95% CI: 5,5-6,82) respectively) compliance rates. For patients with moderate compliance rate, the most significant and almost equivalent factors were the need for lifestyle change — 5,6±3,3 (95% CI: 4,53- 6,71), the need for regular visits — 5,6±3,1 (95% CI: 4,53-6,58) and the need for selfmanagement — 5,6±2,8 (95% CI: 4,63-6,48).Conclusion. The results obtained make it possible to forecast the compliance rate of patients with HTN, and, therefore, direct more efforts to those with a low rate, thereby increasing the effectiveness of AHT

Impaired GABAergic inhibition in the hippocampus of Fmr1 knockout mice.

Many clinical and molecular features of the fragile X syndrome, a common form of intellectual disability and autism, can be modeled by deletion of the Fmr1 protein (Fmrp) in mice. Previous studies showed a decreased expression of several components of the GABAergic system in Fmr1 knockout mice. Here, we used this mouse model to investigate the functional consequences of Fmrp deletion on hippocampal GABAergic inhibition in the CA1-region of the hippocampus. Whole-cell patch-clamp recordings demonstrated a significantly reduced amplitude of evoked inhibitory postsynaptic currents (eIPSCs) and a decrease in the amplitude and frequency of spontaneous IPSCs. In addition, miniature IPSCs were reduced in amplitude and frequency and decayed significantly slower than mIPSCs in controls. Quantitative real-time PCR revealed a significantly lower expression of α2, β1 and δ GABA <sub>A</sub> receptor subunits in the hippocampus of the juvenile mice (P22) compared to wild-type littermates. Correspondingly, we found also at the protein level reduced amounts of α2, β1 and δ subunits in Fmr1 knockout mice. Overall, these results demonstrate that the reduction in several components of the GABAergic system is already present at young age and that this reduction results in measurable abnormalities on GABA <sub>A</sub> receptor-mediated phasic inhibition. These abnormalities might contribute to the behavioral and cognitive deficits of this fragile X mouse model

Proper synaptic vesicle formation and neuronal network activity critically rely on syndapin I

Synaptic transmission relies on effective and accurate compensatory endocytosis. F-BAR proteins may serve as membrane curvature sensors and/or inducers and thereby support membrane remodelling processes; yet, their in vivo functions urgently await disclosure. We demonstrate that the F-BAR protein syndapin I is crucial for proper brain function. Syndapin I knockout (KO) mice suffer from seizures, a phenotype consistent with excessive hippocampal network activity. Loss of syndapin I causes defects in presynaptic membrane trafficking processes, which are especially evident under high-capacity retrieval conditions, accumulation of endocytic intermediates, loss of synaptic vesicle (SV) size control, impaired activity-dependent SV retrieval and defective synaptic activity. Detailed molecular analyses demonstrate that syndapin I plays an important role in the recruitment of all dynamin isoforms, central players in vesicle fission reactions, to the membrane. Consistently, syndapin I KO mice share phenotypes with dynamin I KO mice, whereas their seizure phenotype is very reminiscent of fitful mice expressing a mutant dynamin. Thus, syndapin I acts as pivotal membrane anchoring factor for dynamins during regeneration of SVs. The EMBO Journal (2011) 30, 4955-4969. doi: 10.1038/emboj.2011.339; Published online 16 September 201