Gamma aminobutyric acid uptake,release, and effect on36Cl−-influx in bovine pineal gland

Summary Two apparent affinities for Na⁺ -dependent,³H-GABA uptake were found in bovine pineal fragments in vitro i.e., a high affinity uptake (Km=37±5 M) and a low affinity uptake (Km=435±50 uM). GABA or the neuronal and glial GABA uptake inhibitor nipecotic acid was significantly more effective than the inhibitor of the GABA glial uptake -alanine to decrease pineal³H-GABA uptake. High K⁺ concentration released³H-GABA in superfused bovine pineals, no differences in³H-GABA release among fragments taken from medial, proximal or distal pineal regions being apparent. Superfusion of pineal fragments in the absence of Ca²⁺ but in the presence of EGTA, Mg²⁺ or verapamil decreased significantly³H-GABA release induced by K⁺. In every case a Ca²⁺ -independent pineal GABA release was found. Preincubation with GABA or nipecotic acid, but not with -alanine, blunted subsequent³H-GABA release. GABA increased³⁶Cr^–-influx in pineal homogenates, an effect blocked by picrotoxin. Incubation of pineal homogenates in the presence of aminooxyacetic acid decreased V_max of glutamic acid decarboxylase, without modifying its Km. These results are compatible with a transmitter or modulator role of GABA in bovine pineal gland.     

Rural−urban differences in Austrian suicides

Objectives The answer to the question whether suicide rates are higher in urban than in rural areas may have changed over the years. This study analyzes the longitudinal trends of rural and urban suicides in Austria from 1970 to 2005. The most recent decade, 1995–2005 was also investigated cross-sectionally in terms of age groups, gender, suicide methods and family status. Methods Official suicide statistics were calculated in a Poisson regression model to determine trends in suicide rates according to gender in rural and urban regions as well as the ratios of rural- to urban-suicide rates. Population density levels were used as a measure of urbanization. Differences in suicide rates across the rural–urban categories were investigated in terms of genders, age groups, suicide methods and family status using Spearman correlations. Results The ratio of rural to urban suicide rates has continuously increased in both genders over the past 35 years, indicating a growing risk in rural areas. Suicide methods used in rural and urban areas vary significantly and suicide rates among men, but not women, were found to decrease with increasing urbanicity. Conclusion In line with recent findings from other western countries, we showed a growing gap between rural and urban suicide rates. This suggests a need for rural-specific suicide prevention efforts, especially aimed at the male rural population.     

Association of monocyte chemoattractant protein-1 −2518A>G polymorphism with occurrence, severity, and outcome in ischemic stroke

Monocyte chemoattractant protein-1 (MCP-1) is implicated in promoting atherosclerotic diseases, including stroke. Therefore, several studies have investigated the association between variants of the MCP-1 gene and risk of atherosclerotic diseases. We sought to determine the occurrence of MCP-1 ?2518A>G polymorphism in patients with ischemic stroke (IS), and studied its association with the severity of disease and functional outcome after an acute IS. One hundred and forty-five consecutive patients with first ever IS and 145 age- and sex-matched control subjects were recruited. Stroke severity and functional outcome were assessed on admission and at one month post-stroke, respectively. Genotyping for the MCP-1 ?2518A>G polymorphism was performed by a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). No significant difference in the frequency of MCP-1 ?2518A>G genotypes between IS patients and controls was found, with OR = 0.69 (95 % CI 0.46–1.04, P = 0.08). Moreover, carriage of the G allele was not associated with stroke severity (Scandinavian stroke scale score 33.1 vs. 32.5, respectively, P = 0.71), or poor outcome at 1 month post-stroke (63.9 vs. 59.7 %, respectively, P = 0.61). In conclusion, we were unable to demonstrate a significant association of the MCP-1 ?2518A>G gene polymorphism with IS occurrence, severity or functional outcome in a Caucasian population. However, larger studies are necessary to fully elucidate the role of this polymorphism in IS.     

TLR9 −1237 T/C and TLR2 −194 to −174 del polymorphisms and the risk of Parkinson’s disease in the Greek population: a pilot study

Toll-like receptors (TLRs) are important mediators of inflammatory responses by recognition of many pathogen-related molecules and endogenous proteins related to immune activation. Accumulating data have recently pointed out the role of neuroinflammation in Parkinson’s disease (PD) pathogenesis. In the present study, we investigated the potential role of the TLR9 ?1237 T/C and TLR2 ?194 to ?174 del polymorphisms in PD. We studied a total of 333 individuals, 215 Greek patients with sporadic PD and 118 control subjects, using a polymerase chain reaction–restriction fragment length polymorphism method. No statistically significant differences were found between PD patients and control subjects for the TLR9 ?1237 T/C genotypes or alleles. Regarding the TLR2 ?196 to ?174 del polymorphism, the del/del genotype and the del allele were overrepresented in the PD group compared to controls, however, this result did not reach statistical significance (P = 0.087). Further studies investigating the TLR-inflammatory background of PD are awaited to provide important insight into the aetiology of the disease.     

Cross-cultural gene− environment interactions in depression,post-traumatic stress disorder,and the cortisol awakening response: FKBP5 polymorphisms and childhood trauma in South Asia

Abstract

Despite increased attention to global mental health, psychiatric genetic research has been dominated by studies in high-income countries, especially with populations of European descent. The objective of this study was to assess single nucleotide polymorphisms (SNPs) in the FKBP5 gene in a population living in South Asia. Among adults in Nepal, depression was assessed with the Beck Depression Inventory (BDI), post-traumatic stress disorder (PTSD) with the PTSD Checklist-Civilian Version (PCL-C), and childhood maltreatment with the Childhood Trauma Questionnaire (CTQ). FKBP5 SNPs were genotyped for 682 participants. Cortisol awakening response (CAR) was assessed in a subsample of 118 participants over 3 days. The FKBP5 tag-SNP rs9296158 showed a main effect on depressive symptoms (p = 0.03). Interaction of rs9296158 and childhood maltreatment predicted adult depressive symptoms (p = 0.02) but not PTSD. Childhood maltreatment associated with endocrine response in individuals homozygous for the A allele, demonstrated by a negative CAR and overall hypocortisolaemia in the rs9296158 AA genotype and childhood maltreatment group (p < 0.001). This study replicated findings related to FKBP5 and depression but not PTSD. Gene–environment studies should take differences in prevalence and cultural significance of phenotypes and exposures into account when interpreting cross-cultural findings.     

Interleukin-18 −137 G/C and −607 C/A polymorphisms and Alzheimer’s disease risk: a meta-analysis

The ?137 G/C and ?607 C/A polymorphisms in interleukin-18 (IL-18) gene have been reported to be associated with Alzheimer’s disease (AD) risk, but the results are inconclusive. Considering a single study may lack the power to provide reliable conclusion, we performed a meta-analysis to investigate the association between the IL-18 ?137 G/C and ?607 C/A polymorphisms and AD susceptibility. A comprehensive literature search of PubMed, Embase, China National Knowledge Infrastructure (CNKI) and Wanfang databases were conducted before September 1, 2015. The pooled odds ratio (OR) with 95 % confidence intervals (CIs) were calculated. Five eligible studies with a total of 1536 subjects were finally included in this meta-analysis. For the IL-18 ?137 G/C polymorphism, a significantly decreased risk was detected in patients carrying the C allele of ?137 G/C in all study subjects in allele model (C vs. G: OR = 0.816, 95 % CI = 0.680–0.980, p = 0.029). Moreover, stratification by ethnicity indicated markedly association between the ?137 G/C C allele and AD risk in Asians. For the IL-18 ?607 C/A polymorphism, a significantly decreased risk was found in patients carrying the A allele of ?607 C/A in all study subjects in dominant model (AA + CA vs. CC: OR = 0.696, 95 % CI = 0.529–0.915, p = 0.010). However, the results suggested no significant association between the ?607 C/A polymorphism and AD susceptibility when stratified by ethnicity. Our present meta-analysis suggests that the C allele carrier of IL-18 ?137 G/C was associated with decreased risk for AD in Asians. Further well-designed case–control studies with larger sample size and more ethnic groups are needed to confirm these conclusions.     

Association of Endothelial Nitric Oxide Synthase Gene Polymorphisms (894G/T, −786T/C,G10T) and Clinical Findings in Patients with Migraine

Migraine is a common neurological disorder characterized by recurrent attacks, unilateral head pain, and related symptoms. The aim of this study was to investigate three endothelial nitric oxide synthase (eNOS) polymorphisms in 176 patients with migraine and 123 healthy individuals. Clinical and biochemical parameters were investigated. Genetic analysis was performed using the polymerase chain reaction–restriction fragment length polymorphism method. The differences between migraine cases and the control group were significant for two polymorphisms (?786T/C and 894G/T) (p = 0.000). Homocysteine and body mass index (BMI) were significantly higher in the migraine group than in the control group (p = 0.001 and p = 0.000). The relation between ?786T/C genotype and BMI and allodynia was significant. TC heterozygotes and CC homozygotes were significantly higher in the migraine group than in the control group (OR 2.843 and 95 % CI 1.681–4.808 and OR 3.729 and 95 % CI 1.784–7.792, respectively). The 894G/T genotype was correlated with BMI, pain intensity, age at the onset of migraine, nausea, tension, compression, and allodynia. For this polymorphism, GT heterozygotes and TT homozygotes were significantly higher in the migraine group than in the control group (OR 3.027 and 95 % CI 1.830–5.008 and OR 3.221 and 95 % CI 1.223–8.484, respectively). The G10T genotype was correlated with attack duration and age at the onset of migraine (p = 0.008 and p = 0.040). eNOS polymorphisms may be useful markers for assessing migraine risk and clinical diagnosis.     

CD4+CD28− lymphocytes and ischaemic stroke. Part I: CD4+CD28− lymphocytes and common carotid artery intima-media thickness

Background and purposeMore and more data point to the involvement of the CD4⁺CD28^? lymphocyte subpopulation in the pathogenesis of ischaemic stroke. This paper attempts to answer the question of whether an increase in the percentage of CD4⁺CD28^? lymphocytes in the blood may be associated with carotid artery intima-media thickness (IMT).Material and methodsThe study involved a group of 109 patients, aged 45 to 65 years, including 42 patients with first-ever ischaemic stroke, experiencing symptoms resulting from disturbances of the anterior area of cerebral circulation, arterial hypertension and/or type 2 diabetes mellitus (group 1). Group 2 consisted of 34 patients with above-mentioned risk factors but without ischaemic stroke. The control group comprised 33 healthy individuals. Distribution of sex and mean age was comparable. The IMT of carotid arteries was measured by ultrasonography. Flow cytometry was applied to determine the percentage of CD4⁺CD28^? lymphocytes in the peripheral blood.ResultsThe IMT was significantly greater in patients with stroke than in patients without stroke. No significant correlation was found between the proportion of CD4⁺CD28^? lymphocytes in the blood and the IMT of carotid arteries.ConclusionsThe significant proportion of CD4⁺CD28^? lymphocytes in patients with ischaemic stroke points to the involvement of the cells in the pathogenesis of stroke. The CD4⁺CD28^? lymphocytes are not involved in the pathomechanism of common carotid arteries IMT thickening in this group of patients.     

Development of O4+/O1− immunopanned pro-oligodendroglia in vitro

In this study, O4+/O1− pro-oligodendroglia isolated by immunopanning from cerebral hemispheres of P3–P5 rats were evaluated during their maturation in culture. Immunopanning yielded 3–4×10⁵ cells/cerebrum, with 98% O4+ and 6% O1+. There was heterogeneity in the morphologies of immunopanned cells ranging from simple bipolar cells to more complex multipolar cells. As a first step in determining potential differentiative responses of mature oligodendroglia, we examined glial fibrillary acidic protein (GFAP) expression in response to fetal bovine serum (FBS) by cultures established from O4+/O1− immunopanned cells grown for 1, 14, or 21 days, exposed to 20% FBS for 6–7 days and fixed and immunostained on days 7, 21 or 28 in culture (DIC). When immunopanned cells were exposed to FBS following 1 day in serum-free medium, 88% expressed GFAP and when immunopanned cells were cultured for 14 days prior to FBS exposure, 78% expressed GFAP. By contrast, when cells were cultured for 21 days prior to FBS exposure (when a majority of the cells expressed O1 and myelin basic protein (MBP)), only 19% of the cells expressed GFAP (p<0.001). Cells that were O4+/GFAP− even in the presence of FBS often exhibited a mature oligodendroglial morphology. Among immunopanned cells that responded to FBS by expression of GFAP, both process-bearing (similar to type 2 astroglia) and flattened, polygonal (similar to type 1 astroglia) GFAP+ cells were observed. These results confirm the utility of immunopanning for the isolation of pro-oligodendroglia and demonstrate that oligodendroglia that develop in vitro from O4+/O1− immunopanned cells become resistant to GFAP induction by FBS.     

Effects of GABAA-ergic ligands on Cl− transport induced by the Cl−, HCO 3 − -ATPase from carp (Cyprinus carpio L.) brain reconstituted in proteoliposomes

A Cl^?, HCO ₃ ^? -ATPase isolated from the plasma membranes of fish brain was reconstituted in artificial proteoliposomes. The original sensitivity to GABA_A-ergic ligands was preserved in the reconstituted enzyme. GABA was shown to activate the liposome-embedded enzyme in a concentration range from 10 to 100 μM while picrotoxin (100 μM) removed this activating affect. A Cl^?-sensitive fluorescent probe (MEQ) was used to study the ATP-dependent Cl^? transport through the membrane of the proteoliposomes. ATP (in concentrations of 1.5 mM and higher) was found to induce Cl^?-transport into the proteoliposomes. This ATP-dependent transport was increased in the presence of GABA (100 μM), while the latter effect could also be blocked by picrotoxin (100 μM). It was concluded that the Cl^?, HCO ₃ ^? -ATPase could be involved in ATP-dependent Cl^?-transport and regulated by activators and blockers of GABA_A receptors.     

CD4+CD28− lymphocytes and cerebral ischaemic stroke. Part II: CD4+CD28− lymphocytes and carotid artery atherosclerotic plaque characteristics

Background and purposeCD4⁺CD28^? lymphocytes can directly contribute to the instability of atherosclerotic plaque. This paper attempts to answer the question of the potential influence of the CD4⁺CD28^? lymphocyte population on the ultrasound image of atherosclerotic plaque in the common carotid artery (CCA) wall.Material and methodsThe study involved a group of 109 patients, aged 45 to 65 years, including 42 patients with first-ever ischaemic stroke, experiencing symptoms resulting from disturbances of the anterior area of cerebral circulation, arterial hypertension and/or type 2 diabetes mellitus (group 1). Group 2 consisted of 34 patients with mentioned risk factors, without ischaemic stroke. The control group comprised 33 healthy individuals. The percentage of CD4⁺CD28^? lymphocytes was assessed with flow cytometry.ResultsA significant difference in the incidence of heterogeneous plaques was noted between groups 1 and 3 (p = = 0.0023) as well as between group 2 and 3 (p = 0.0005), whereas groups 1 and 2 did not differ from each other. The proportion of CD4⁺CD28^? lymphocytes was similar in groups 1 and 2 (p = 0.97), but it differed between groups 1 and 3 (p < 0.0001) and between groups 2 and 3 (p < 0.001). A correlation was found between the proportion of CD4⁺CD28^? lymphocytes in the blood and the number of CCA atherosclerotic plaques (Rs = 0.191, p = 0.046). The proportion of CD4⁺CD28^? lymphocytes in peripheral blood did not correlate with the ultrasound types of atherosclerotic plaques. No correlation between the proportion of CD4⁺CD28^? lymphocytes and the area of atherosclerotic plaques was found.ConclusionsThe correlation between the proportion of CD4⁺CD28^? lymphocytes and the number of atherosclerotic plaques within the CCA suggests that the cells are involved in the mechanism of carotid plaque formation. There is no proof of the involvement of the above-mentioned cells in the mechanism of plaque destabilization in those arteries.     

Interaction analysis between 5-HTTLPR and TNFA −238/−308 polymorphisms in schizophrenia

Summary. This study investigated the potential interaction between the polymorphisms of serotonin transporter gene (SLC6A4, a 44 base pair insertion/deletion in the promoter region, 5-HTTLPR) and tumor necrosis factor-α gene (TNFA; −238G/A and −308G/A polymorphisms) on the development of schizophrenia, as well as the interaction of the three polymorphisms in relation to symptomatology, family history, onset age and antipsychotic treatment response. Genomic DNA analyses with polymerase chain reaction (PCR) was used for the genotyping. One hundred and fifty-two (152) patients with schizophrenia and 152 normal controls participated in the study. Any associations between the individual polymorphism and schizophrenia were not found. However, marginal association between subjects with both TNFA −238 A allele (genotype AA plus AG) and 5-HTTLPR s allele (ss plus sl) and presence of family history was found (p = 0.023; p = 0.026). The subjects with TNFA −308 AG genotype showed higher change in PANSS total score (p = 0.028). No significant interaction effect between 5-HTTLPR and TNFA −238/−308 polymorphisms either on the development of schizophrenia or on antipsychotics treatment response and psychopathology was found, although a significant interaction effect for subjects carrying TNFA −238 AG and −308 AA genotypes on a positive family history was observed (p = 0.017). These results suggest that the interaction effects between 5-HTTLPR and TNFA −238/−308 polymorphisms gives no significant contribution to the susceptibility to schizophrenia, and is not associated with clinical variables, antipsychotic treatment response and psychopathological features, except for family history of disease, at least in the Korean population.     

Hypoalbuminemia in early onset dentatorubral−pallidoluysian atrophy due to leakage of albumin in multiple organs

We delineate a complication of hypoalbuminemia in dentatorubral?pallidoluysian atrophy (DRPLA), which we have found to be common in this disorder. In addition, we explored the pathogenesis of this phenomenon through clinical and histological examinations. Clinical course and laboratory findings of nine patients with childhood-onset DRPLA (aged 6–49 years; CAG repeat length 62–93) were retrospectively reviewed. Autopsied specimens from three patients were examined by histopathological and immunohistochemical analyses. Eight DRPLA patients showed hypoalbuminemia <3.5 g/dl in the initial stages of the disease (age, 2–32 years), which correlated with the CAG repeat length in each patient. Disease worsened in six patients, often triggered by febrile infections and accompanied by increased urinary protein excretion. One patient showed increased fecal α1-antitripsin while another showed accumulation of radioactive albumin in the urinary and gastrointestinal tracts after intravenous infusion. Immunohistochemistry revealed albumin-containing monocytes and astrocytes in the perivascular areas of the cerebral white matter. Fluid collection in the glomerular capillaries was noted. Immunolabeling using antibodies against the expanded polyglutamine (polyQ) polypeptide was positive in cerebral cortical neurons, hepatocytes, renal collecting ducts, and glomerular podocytes, which act as filtration barrier against serum proteins. Serum albumin appears to easily leak from blood vessels in certain visceral organs in DRPLA during later stages of the illness, particularly in the kidneys of patients with largely expanded CAG repeats. We hypothesize that the accumulation of the DRPLA gene product with expanded polyQ sequences in the podocytes results in the dysfunction of the glomerular filtration barrier.