Peroxynitrite oxidizes erythrocyte membrane band 3 protein and diminishes its anion transport capacity

We describe an altered membrane band 3 protein-mediated anion transport in erythrocytes exposed to peroxynitrite, and relate the loss of anion transport to cell damage and to band 3 oxidative modifications. We found that peroxynitrite down-regulate anion transport in a dose dependent relation (100–300 μmoles/l). Hemoglobin oxidation was found at all peroxynitrite concentrations studied. A dose-dependent band 3 protein crosslinking and tyrosine nitration were also observed. Band 3 protein modifications were concomitant with a decrease in transport activity. ( ? )-Epicatechin avoids band 3 protein nitration but barely affects its transport capacity, suggesting that both processes are unrelated. N-acetyl cysteine partially reverted the loss of band 3 transport capacity. It is concluded that peroxynitrite promotes a decrease in anion transport that is partially due to the reversible oxidation of band 3 cysteine residues. Additionally, band 3 tyrosine nitration seems not to be relevant for the loss of its anion transport capacity.     

Sulfate influx on band 3 protein of equine erythrocyte membrane (Equus caballus) using different experimental temperatures and buffer solutions

The aim of this study was to assess the anion transport in equine erythrocytes through the measurement of the sulfate uptake operating from band 3 using different experimental temperatures and buffer solutions. Blood samples of six clinically healthy horses were collected via jugular vein puncture, and an emochrome‐citometric examination was performed. The blood was divided into four aliquots and by centrifugation and aspiration the plasma and buffy coat were carefully discarded. The red blood cells were washed with an isosmotic medium and centrifuged. The obtained cell suspensions were incubated with two different experimental buffer solutions (buffer A: 115 mM Na2SO4, 10 mM NaCl, 20 mM ethylenediaminetetraacetic acid, 30 mM glucose; and buffer B: 115 mM Na2SO4, 10 mM NaCl, 20 mM ethylenediaminetetraacetic acid, 30 mM MgCl2) in a water bath for 1 h at 25 °C and 37 °C. Normal erythrocytes, suspended at 3% hematocrit, were used to measure the influx by absorption spectrophotometry at 425 nm wavelength. Unpaired Student's t‐test showed a statistically significant decrease (P < 0.01) of rate constants in equine erythrocytes at 25 °C versus 37 °C using both experimental buffer solutions. Comparing the buffer A with buffer B unpaired Student's t‐test showed statistically lower values (P < 0.0001) for A solution versus B solution both at 25 °C and at 37 °C. The greater inhibition of SO₄⁼ influx measured in equine erythrocytes indicates the increased formation of the sulfydryl bonds in band 3 and the modulation of the sulfydryl groups, culminating in the conformational changes in band 3. Copyright © 2012 John Wiley & Sons, Ltd.     

The neurobiology of adaptation to seasons: Relevance and correlations in bipolar disorders

ABSTRACT

Bipolar disorders (BDs) are severe and common psychiatric disorders. BD pathogenesis, clinical manifestations and relapses are associated with numerous circadian rhythm abnormalities. In addition, infradian fluctuations of mood, social activity, weight and sleep patterns are very frequent in BD. Disease course with a seasonal pattern (SP) occurs in approximately 25% of depressive and 15% of manic episodes, which is coupled to a more severe disease symptomatology. The pathophysiological mechanisms of seasonal effects in BD await clarification, with likely important clinical consequences. This review aims at synthesizing available data regarding the underlying pathophysiological mechanisms of seasonality in BD patients, with implications for future research directions in the study of seasonality in BD. Three factors are suggested to play significant roles in BD with SP, namely the suprachiasmatic nuclei, as well as the melatonergic and photoperiodism systems. It is proposed that BD with SP may be considered as a complex disorder resulting from the interaction of clock gene vulnerabilities and biological clock neuroplasticity, with environmental factors, such as the response to light. Light seems to play a key role in BD with SP, mainly due to two seasonal signaling pathways: a light to cortex serotonin transporter pathway, as well as a pathway connecting light to melatonin synthesis. This provides a theoretical framework for BD with SP, including for future research and clinical management. The review proposes that future research should explore markers of seasonality in BD, such as plasma melatonin, sleep–wake rhythms (with actigraphy) and genetic or epigenetic variants within the melatonin synthesis pathway. The role of light in driving BD with SP is an active area of research. Seasonality may also be intimately linked to wider aspects of BD, including via interactions with the gut microbiome, the gut–liver axis, cholesterol regulation, aspects of metabolic syndrome, vitamin D, decreased longevity, suicide risk and medication treatment targets. Further research on the role of seasonality in BD is likely to clarify the etiology, course and treatment of BD more widely.     

Dependence of Subjective Traverse Length on Velocity of Moving Tactile Stimuli

Two series of experiments were performed to assess the effects of stimulus velocity on human subjects' perception of the distance traversed by a moving tactile stimulus. In all experiments, constant-velocity stimuli were applied to the dorsal surface of the left forearm; velocities ranging between 1.0 and 256 cm/sec were used. In some experiments the stimuli moved from distal to proximal over the skin, and in others they moved from proximal to distal. The length of skin contacted by the moving stimulus was defined by a plate having an aperture of 4.0 × 0.5 cm.

In the first series of experiments, subjects were required to compare the distance traversed by a test stimulus delivered 2 sec after a standard stimulus, and also to report the on-locus and the off-locus of the brushing stimulus. In the second series of experiments, the subjects rated the perceived distance on the skin using a free-magnitude-estimation procedure. The data from both series of experiments defined the same relationship between stimulus velocity and perceived stimulus distance. More specifically, although the length of skin contacted by the stimulus was the same at all velocities, subjects' estimates of stimulus distance decreased with increasing stimulus velocity. In addition, the function relating estimates of stimulus distance to velocity was flat for velocities between 5 and 20 cm/sec, but possessed an appreciable negative slope at lower and higher velocities.

It is interesting that the plateau of the relationship between perceived stimulus distance and velocity occurred within the range of velocities that human subjects employ to scan textured surfaces; it also corresponded precisely with the range of stimulus velocities at which the directional sensitivity of somatosensory cortical neurons and human subjects is optimal.     

Growth Arrest Specific 1 (GAS1) Is Abundantly Expressed in the Adult Mouse Central Nervous System

Growth arrest specific 1 (GAS1) is a pleiotropic protein that induces apoptosis and cell arrest in different tumors, but it is also involved in the development of the nervous system and other tissues and organs. This dual ability is likely caused by its capacity to interact both by inhibiting the intracellular signaling cascade induced by glial cell-line derived neurotrophic factor and by facilitating the activity of the sonic hedgehog pathway. The presence of GAS1 mRNA has been described in adult mouse brain, and here we corroborated this observation. We then proceeded to determine the distribution of the protein in the adult central nervous system (CNS). We detected, by western blot analysis, expression of GAS1 in olfactory bulb, caudate-putamen, cerebral cortex, hippocampus, mesencephalon, medulla oblongata, cerebellum, and cervical spinal cord. To more carefully map the expression of GAS1, we performed double-label immunohistochemistry and noticed expression of GAS1 in neurons in all brain areas examined. We also observed expression of GAS1 in astroglial cells, albeit the pattern of expression was more restricted than that seen in neurons. Briefly, in the present article, we report the widespread distribution and cellular localization of the GAS1 native protein in adult mammalian CNS.     

FERM domain promotes resveratrol-induced apoptosis in endothelial cells via inhibition of NO production

Focal adhesion kinase (FAK) consists of an N-terminal band 4.1; ezrin, radixin, moesin (FERM) domain; tyrosine kinase domain; and C-terminal FA targeting domain. Here we show that ectopically expressed FERM is largely located in the cytosolic fraction under quiescent conditions. We further found that this ectopically expressed FERM domain aggravates endothelial cell apoptosis triggered by 100 μM resveratrol, whereas FERM had no effect on apoptosis induced by TNF-α. We determined that resveratrol at low doses (<20 μM) promotes phosphorylation (S1177) of eNOS via an AMPK-dependent pathway. The presence of the FERM domain blocked this resveratrol-stimulated eNOS phosphorylation and NO production. Thus, the pro-apoptotic activity of cytosolic FERM domain is at least partially mediated by down-regulation of NO, a critical cell survival factor. Consistently, we found that the apoptosis induced by cytosolic FERM in the presence of resveratrol was reversed by an NO donor, SNAP. In conclusion, FERM located in the cytosolic fraction plays a pivotal role in aggravating cell apoptosis through diminishing NO production.     

颈动脉斑块LRNC特征可诊断2型糖尿病患者急性脑梗死的风险

2型糖尿病可能加重颈动脉斑块的易损性并增加缺血性中风的风险,关于2型糖尿病患者伴有颈动脉斑块特征的急性中风亚型鲜有研究报道。本研究旨在探讨2型糖尿病患者颈动脉斑块特征与MRI确定的急性脑梗死病变特征之间的关系。本研究以颈内动脉区急性脑血管病患者为研究对象,所有患者分为2型糖尿病组和非2型糖尿病组,分别行颈动脉和脑部MRI扫描,测定同侧颈动脉斑块的形态和特征,以及颅内和颅外颈动脉狭窄。基于中风亚型和急性脑梗塞病变模式对患者进行评估。研究结果表明,与非2型糖尿病患者相比,2型糖尿病患者颈动脉型IV-VI病变的患病率更高,斑块负荷更大,以及富脂质坏死核(LRNC)更大。在有症状的颈动脉LRNC患者中,与非2型糖尿病组相比,2型糖尿病组颈内动脉区出现较多的伴有大穿孔动脉梗塞形态和较大的急性脑梗塞。LRNC%>23.5%的颈动脉斑块是2型糖尿病患者存在颈动脉狭窄的急性脑梗塞病变的独立危险因素。颈动脉斑块特征的量化,尤其是MRI诊断的富脂质坏死核对中风风险具有潜在应用价值。