STIM 1/Orai 1 contributes to sex differences in vascular responses to calcium in spontaneously hypertensive rats

Sex differences in Ca2+-dependent signalling and homoeostasis in the vasculature of hypertensive rats are well characterized. However, sex-related differences in SOCE (store-operated Ca2+ entry) have been minimally investigated. We hypothesized that vascular protection in females, compared with males, reflects decreased Ca2+ mobilization due to diminished activation of Orai 1/STIM 1 (stromal interaction molecule I). In addition, we investigated whether ovariectomy in females affects the activation of the Orai 1/STIM 1 pathway. Endothelium-denuded aortic rings from male and female SHRSP (stroke-prone spontaneously hypertensive rats) and WKY (Wistar Kyoto) rats and from OVX (ovariectomized) or sham female SHRSP and WKY rats were used to functionally evaluate Ca2+ influx-induced contractions. Compared with females, aorta from male SHRSP displayed: (i) increased contraction during the Ca2+-loading period; (ii) similar transient contraction during Ca2+ release from the intracellular stores; (iii) increased activation of STIM 1 and Orai1, as shown by the blockade of STIM 1 and Orai1 with neutralizing antibodies, which reversed the sex differences in contraction during the Ca2+-loading period; and (iv) increased expression of STIM I and Orai I. Additionally, we found that aortas from OVX-SHRSP showed increased contraction during the Ca2+-loading period and increased Orai1 expression, but no changes in the SR (sarcoplasmic reticulum)-buffering capacity or STIM I expression. These findings suggest that augmented activation of STIM 1/Orai 1 in aortas from male SHRSP represents a mechanism that contributes to sex-related impaired control of intracellular Ca2+ levels. Furthermore, female sex hormones may negatively modulate the STIM/Orai 1 pathway, contributing to vascular protection observed in female rats.

Redox modulation of STIM-ORAI signaling.

STIM1 and ORAI1 constitute the core machinery of the ubiquitous store-operated calcium entry pathway and loss of function in these proteins is associated with severe immune and muscular disorders. Other isoforms-STIM1L, STIM2, ORAI2 and ORAI3 exhibit varied expression levels in different cell types along with several other interaction partners and thereby play different roles to facilitate, regulate and fine-tune the calcium entry. STIM proteins convey the Ca(2+) store-depletion message to the PM and thereby participate in refilling of the ER by physically interacting with the Ca(2+)-selective ORAI channels at the PM. STIM and ORAI are exposed to oxidative modifications in the ER, the cytosol, and at the cell surface, and redox-mediated alterations in STIM/ORAI coupling might contribute to autoimmune disorders and cancer progression. This review discusses the redox reactivity of cysteine residues in STIM and ORAI isoforms, focusing on the oxidative modifications of STIM and ORAI proteins by which STIM-ORAI signaling can be modulated.

Cow's milk allergy and the development of tolerance

Aims: To gain insight on the immunological processes behind cow’s milk allergy (CMA) and the development of oral tolerance. To furthermore investigate the associations of HLA II and filaggrin genotypes with humoral responses to early oral antigens. Methods: The study population was from a cohort of 6209 healthy, full-term infants who in a double-blind randomized trial received supplementary feeding at maternity hospitals (mean duration 4 days): cow’s milk (CM) formula, extensively hydrolyzed whey formula or donor breast milk. Infants who developed CM associated symptoms that subsided during elimination diet (n=223) underwent an open oral CM challenge (at mean age 7 months). The challenge was negative in 112, and in 111 it confirmed CMA, which was IgE-mediated in 83. Patients with CMA were followed until recovery, and 94 of them participated in a follow-up study at age 8-9 years. We investigated serum samples at diagnosis (mean age 7 months, n=111), one year later (19 months, n=101) and at follow-up (8.6 years, n=85). At follow-up, also 76 children randomly selected from the original cohort and without CM associated symptoms were included. We measured CM specific IgE levels with UniCAP (Phadia, Uppsala, Sweden), and β-lactoglobulin, α-casein and ovalbumin specific IgA, IgG1, IgG4 and IgG levels with enzyme-linked immunosorbent assay in sera. We applied a microarray based immunoassay to measure the binding of IgE, IgG4 and IgA serum antibodies to sequential epitopes derived from five major CM proteins at the three time points in 11 patients with active IgE-mediated CMA at age 8-9 years and in 12 patients who had recovered from IgE-mediated CMA by age 3 years. We used bioinformatic methods to analyze the microarray data. We studied T cell expression profile in peripheral blood mononuclear cell (PBMC) samples from 57 children aged 5-12 years (median 8.3): 16 with active CMA, 20 who had recovered from CMA by age 3 years, 21 non-atopic control subjects. Following in vitro β-lactoglobulin stimulation, we measured the mRNA expression in PBMCs of 12 T-cell markers (T-bet, GATA-3, IFN-γ, CTLA4, IL-10, IL-16, TGF-β, FOXP3, Nfat-C2, TIM3, TIM4, STIM-1) with quantitative real time polymerase chain reaction, and the protein expression of CD4, CD25, CD127, FoxP3 with flow cytometry. To optimally distinguish the three study groups, we performed artificial neural networks with exhaustive search for all marker combinations. For genetic associations with specific humoral responses, we analyzed 14 HLA class II haplotypes, the PTPN22 1858 SNP (R620W allele) and 5 known filaggrin null mutations from blood samples of 87 patients with CMA and 76 control subjects (age 8.0-9.3 years). Results: High IgG and IgG4 levels to β-lactoglobulin and α-casein were associated with the HLA (DR15)-DQB1*0602 haplotype in patients with CMA, but not in control subjects. Conversely, (DR1/10)-DQB1*0501 was associated with lower IgG and IgG4 levels to these CM antigens, and to ovalbumin, most significantly among control subjects. Infants with IgE-mediated CMA had lower β -lactoglobulin and α-casein specific IgG1, IgG4 and IgG levels (p<0.05) at diagnosis than infants with non-IgE-mediated CMA or control subjects. When CMA persisted beyond age 8 years, CM specific IgE levels were higher at all three time points investigated and IgE epitope binding pattern remained stable (p<0.001) compared with recovery from CMA by age 3 years. Patients with persisting CMA at 8-9 years had lower serum IgA levels to β-lactoglobulin at diagnosis (p=0.01), and lower IgG4 levels to β-lactoglobulin (p=0.04) and α-casein (p=0.05) at follow-up compared with patients who recovered by age 3 years. In early recovery, signal of IgG4 epitope binding increased while that of IgE decreased over time, and binding patterns of IgE and IgG4 overlapped. In T cell expression profile in response to β –lactoglobulin, the combination of markers FoxP3, Nfat-C2, IL-16, GATA-3 distinguished patients with persisting CMA most accurately from patients who had become tolerant and from non-atopic subjects. FoxP3 expression at both RNA and protein level was higher in children with CMA compared with non-atopic children. Conclusions: Genetic factors (the HLA II genotype) are associated with humoral responses to early food allergens. High CM specific IgE levels predict persistence of CMA. Development of tolerance is associated with higher specific IgA and IgG4 levels and lower specific IgE levels, with decreased CM epitope binding by IgE and concurrent increase in corresponding epitope binding by IgG4. Both Th2 and Treg pathways are activated upon CM antigen stimulation in patients with CMA. In the clinical management of CMA, HLA II or filaggrin genotyping are not applicable, whereas the measurement of CM specific antibodies may assist in estimating the prognosis.

Training-induced changes in the pattern of triceps to biceps activation during reaching tasks after chronic and severe stroke

This exploratory study was undertaken to investigate the mechanisms that contributed to improvements in upper limb function following a novel training program. Surface electromyography (EMG) was used to examine training-induced changes in the pattern of triceps and biceps activation during reaching tasks in stroke survivors with severe paresis in the chronic stage of recovery. The EMG data were obtained in the context of a single blind randomised clinical trial conducted with 42 stroke survivors with minimal upper limb muscle activity and who were more than 6 months post-stroke. Of the 33 participants who completed the study, 10 received training of reaching using a non-robotic upper limb training device, the SMART Arm, with EMG triggered functional electrical stimulation (EMG-stim), 13 received training of reaching using the SMART Arm alone, and 10 received no intervention. Each intervention group engaged in 12 1-h training sessions over a 4-week period. Clinical and laboratory measures of upper limb function were administered prior to training (0 weeks), at completion (4 weeks) and 2 months (12 weeks) after training. The primary outcome measure was 'upper arm function' which is Item 6 of the Motor Assessment Scale (MAS). Laboratory measures consisted of two multijoint reaching tasks to assess 'maximum isometric force' and 'maximum distance reached'. Surface EMG was used to monitor triceps brachii and biceps brachii during the two reaching tasks. To provide a comparison with normal values, seven healthy adults were tested on one of the reaching tasks according to the same procedure. Study findings demonstrated a statistically significant improvement in upper limb function for stroke participants in the two training groups compared to those who received no training however no difference was found between the two training groups. For the reaching tasks, all stroke participants, when compared to normal healthy adults, exhibited lower triceps and biceps activation and a lower ratio of triceps to biceps activation. Following training, stroke participants demonstrated increased triceps activation and an increased ratio of triceps to biceps activation for the task that was trained. Better performance was associated with greater triceps activation and a higher ratio of triceps to biceps activation. The findings suggest that increased activation of triceps as an agonist and an improved coordination between triceps and biceps could have mediated the observed changes in arm function. The changes in EMG activity were small relative to the changes in arm function indicating that factors, such as the contribution of other muscles of reaching, may also be implicated.

Primary motor cortex involvement in initial learning during visuomotor adaptation

Human motor behaviour is continually modified on the basis of errors between desired and actual movement outcomes. It is emerging that the role played by the primary motor cortex (M1) in this process is contingent upon a variety of factors, including the nature of the task being performed, and the stage of learning. Here we used repetitive TMS to test the hypothesis that M1 is intimately involved in the initial phase of sensorimotor adaptation. Inhibitory theta burst stimulation was applied to M1 prior to a task requiring modification of torques generated about the elbow/forearm complex in response to rotations of a visual feedback display. Participants were first exposed to a 30° clockwise (CW) rotation (Block A), then a 60° counterclockwise rotation (Block B), followed immediately by a second block of 30° CW rotation (A2). In the STIM condition, participants received 20s of continuous theta burst stimulation (cTBS) prior to the initial A Block. In the conventional (CON) condition, no stimulation was applied. The overt characteristics of performance in the two conditions were essentially equivalent with respect to the errors exhibited upon exposure to a new variant of the task. There were however, profound differences between the conditions in the latency of response preparation, and the excitability of corticospinal projections from M1, which accompanied phases of de-adaptation and re-adaptation (during Blocks B and A2). Upon subsequent exposure to the A rotation 24h later, the rate of re-adaptation was lower in the stimulation condition than that present in the conventional condition. These results support the assertion that primary motor cortex assumes a key role in a network that mediates adaptation to visuomotor perturbation, and emphasise that it is engaged functionally during the early phase of learning.

SMART Arm with outcome-triggered electrical stimulation:a pilot randomized clinical trial

The SMART (SensoriMotor Active Rehabilitation Training) Arm is a nonrobotic device designed to allow stroke survivors with severe paresis to practice reaching. It can be used with or without outcome-triggered electrical stimulation (OT-stim) to augment movement. The aim of this study was to evaluate the efficacy of SMART Arm training when used with or without OT-stim, in addition to usual care, as compared with usual care alone during inpatient rehabilitation.

The efficacy of SMART Arm training early after stroke for stroke survivors with severe upper limb disability:a protocol for a randomised controlled trial

Recovery of upper limb function after stroke is poor. The acute to subacute phase after stroke is the optimal time window to promote the recovery of upper limb function. The dose and content of training provided conventionally during this phase is however, unlikely to be adequate to drive functional recovery, especially in the presence of severe motor disability. The current study concerns an approach to address this shortcoming, through evaluation of the SMART Arm, a non-robotic device that enables intensive and repetitive practice of reaching by stroke survivors with severe upper limb disability, with the aim of improving upper limb function. The outcomes of SMART Arm training with or without outcome-triggered electrical stimulation (OT-stim) to augment movement and usual therapy will be compared to usual therapy alone.

La dynamique spatio-temporelle des flux d’oxyde nitreux (N2O) des lacs, rivières, et étangs boréaux

L’oxyde nitreux (N2O), un puissant gaz à effet de serre (GES) ayant plus de 300 fois le potentiel de réchauffement du dioxyde de carbone (CO2), est produit par des processus microbiens du cycle de l’azote (N). Bien que les eaux de surface continentales soient reconnues comme des sites actifs de transformations de l’azote, leur intégration dans les budgets globaux de N2O comporte de nombreuses incertitudes, dont l’absence des lacs dans ces modèles. Le biome boréal est caractérisé par une des plus grandes densités d’eaux douces au monde, pourtant aucune évaluation exhaustive des émissions aquatiques de N2O n’a à date été conduite dans cette région. Dans la présente étude, nous avons mesuré les concentrations de N2O à travers une large gamme de lacs, rivières, et étangs, dans quatre régions boréales du Québec (Canada), et nous avons calculé les flux eau-air résultants. Les flux nets fluctuent entre -23.1 et 177.9 μmol m-2 J-1, avec une grande variabilité inter-système, inter-régionale, et saisonnière. Étonnamment, 40% des systèmes échantillonnés agissaient en tant que puits de N2O durant l’été, et le réseau d’eaux de surfaces d’une des régions était un net consommateur de N2O. Les concentrations maximales de N2O ont été mesurées en hiver dû à l’accumulation de ce gaz sous la glace. Nous avons estimé que l’émission qui en résulte lors de la fonte des glaces représente 20% des émissions annuelles des eaux douces. Parmi les types d’eaux douces échantillonnées, les lacs sont les principaux responsables du flux aquatique net (jusqu’à 90%), et doivent donc être intégrés dans les budgets globaux de N2O. En se basant sur les données empiriques de la littérature, nous avons éstimé l’émission globale de N2O des eaux douces à 0.78 Tg N (N2O) an-1. Ce chiffre est influencé par les émissions des régions de hautes latitudes (tel que le biome boréal) dont les flux nets varient de positif à négatif constituant -9 à 27 % du total.

Increased Activation of Stromal Interaction Molecule-1/Orai-1 in Aorta From Hypertensive Rats A Novel Insight Into Vascular Dysfunction

Disturbances in the regulation of cytosolic calcium (Ca(2+)) concentration play a key role in the vascular dysfunction associated with arterial hypertension. Stromal interaction molecules (STIMs) and Orai proteins represent a novel mechanism to control store-operated Ca(2+) entry. Although STIMs act as Ca(2+) sensors for the intracellular Ca(2+) stores, Orai is the putative pore-forming component of Ca(2+) release-activated Ca(2+) channels at the plasma membrane. We hypothesized that augmented activation of Ca(2+) release-activated Ca(2+)/Orai-1, through enhanced activity of STIM-1, plays a role in increased basal tonus and vascular reactivity in hypertensive animals. Endothelium-denuded aortic rings from Wistar-Kyoto and stroke-prone spontaneously hypertensive rats were used to evaluate contractions because of Ca(2+) influx. Depletion of intracellular Ca(2+) stores, which induces Ca(2+) release-activated Ca(2+) activation, was performed by placing arteries in Ca(2+) free-EGTA buffer. The addition of the Ca(2+) regular buffer produced greater contractions in aortas from stroke-prone spontaneously hypertensive rats versus Wistar-Kyoto rats. Thapsigargin (10 mu mol/L), an inhibitor of the sarcoplasmic reticulum Ca(2+) ATPase, further increased these contractions, especially in stroke-prone spontaneously hypertensive rat aorta. Addition of the Ca(2+) release-activated Ca(2+) channel inhibitors 2-aminoethoxydiphenyl borate (100 mu mol/L) or gadolinium (100 mu mol/L), as well as neutralizing antibodies to STIM-1 or Orai-1, abolished thapsigargin-increased contraction and the differences in spontaneous tone between the groups. Expression of Orai-1 and STIM-1 proteins was increased in aorta from stroke-prone spontaneously hypertensive rats when compared with Wistar-Kyoto rats. These results support the hypothesis that both Orai-1 and STIM-1 contribute to abnormal vascular function in hypertension. Augmented activation of STIM-1/Orai-1 may represent the mechanism that leads to impaired control of intracellular Ca(2+) levels in hypertension. (Hypertension. 2009; 53[part 2]: 409-416.)

Död eller levande? : Vad anser branschfolk i skivbolagen om bolagens positionoch roll i Sverige?

I denna uppsats läggs fokus på skivbranschen och vad folk inom branschen anser omdess funktion och roll.Syftet med uppsatsen är att återge skivbolagens egna tankar och åsikter och jämföradessa med årsredovisningar från branschorganisationer, rapporter, litteratur,vetenskapliga artiklar samt den allmänna uppfattningen hämtad från blogg- ochforuminlägg och tidningsartiklar.Uppsatsen bygger till största delen av intervjuer med totalt sex personer som ärverksamma i branschen från den allra högsta nivån av multinationella bolag till deabsolut minsta.Den bild som framkommer i uppsatsen visar på att den vanligt förekommandeuppfattningen om att skivbolagen idag spelat ut sin roll är felaktig. Skivbolagen hjälperartister med bl.a. marknadsföring, utvecklingen av musik, all administrering samt att dehar de resurser som krävs för att nå ut till den breda massan. Det är dock beroende påvar i karriären man befinner sig och vad artisten själv vill uppnå. För vissa kan detsåledes vara onödigt. Uppsatsen visar även att skivbolagens roll förändrats drastiskt pågrund av bl.a. utbredningen av ny digital teknik. Det är också möjligt att på grund avdessa nya förutsättningar kan rollen förändras ytterligare i framtiden.Skivbolagen arbetar numera på en betydligt bredare front än tidigare. Verksamhetencentreras inte runt den fysiska försäljningen utan förlags- och licensieringsverksamheterär mycket aktuella. Samarbetet mellan små och stora skivbolag påvisas tydligt, en formav samarbete där båda parterna i många fall är beroende av varandra. De mindrebolagen förefaller även vara mer benägna att ta risker i samband med kontraktering avnya akter.

Pengarna som inte kommer fram : Den ekonomiska fördelningsproblematiken i klubbmusik Sverige 2015

Denna uppsats problematiserar hur kollektiva förvaltningsorganisationer fördelar pengar insamlade från offentligt framförd klubbmusik samt undersöker hur etablerade dj’s och producenter ser på dagens system och tror sig påverkas av detta. Med djupintervjuer av fyra personer undersöker uppsatsen även hur dessa ser på rapportering av den spelade musiken, vilka problem som kan uppstå och om det finns tekniska möjligheter att underlätta för korrekt fördelning. Resultaten visar att intervjupersonerna skiljer sig åt i vilken typ av karriär de har och därmed hur man förhåller sig till dagens system. Ingen ställde sig dock positiv till att rapportera in spellistor med vad de spelat som dj’s, man lyfte även fram problemet med att ersätta musik som spelas men ännu inte släppts officiellt. Det framkom också att vi idag har tekniska möjligheter att övervaka vad som spelas genom användandet av akustiska fingeravtryck. Jag diskuterade att det är först när ett sådant system är på plats som vi kan se hur mycket fel dagens pengar fördelas.

A VHDL-based protocol controller for NCAP processors

This work presents the development of an IEEE 1451.2 protocol controller based on a low-cost FPGA that is directly connected to the parallel port of a conventional personal computer. In this manner it is possible to implement a Network Capable Application Processor (NCAP) based on a personal computer, without parallel port modifications. This approach allows supporting the ten signal lines of the 10-wire IEEE 1451.2 Transducer Independent Interface (TII), that connects the network processor to the Smart Transducer Interface Module (STIM) also defined in the IEEE 1451.2 standard. The protocol controller is connected to the STIM through the TII's physical interface, enabling the portability of the application at the transducer and network processor level. The protocol controller architecture was fully developed in VHDL language and we have projected a special prototype configured in a general-purpose programmable logic device. We have implemented two versions of the protocol controller, which is based on IEEE 1451 standard, and we have obtained results using simulation and experimental tests. (c) 2008 Elsevier B.V. All rights reserved.

Network node with wireless and wired interfaces: Nios II processor and uClinux to development of a NCAP embedded (IEEE 1451.1) with two interfaces, wireless (IEEE 1451.5) and wired (IEEE p1451.2)

This paper presents a NCAP embedded on DE2 kit with Nios II processor and uClinux to development of a network gateway with two interfaces, wireless (ZigBee) and wired (RS232) based on IEEE 1451. Both the communications, wireless and wired, were developed to be point-to-point and working with the same protocols, based on IEEE 1451.0-2007. The tests were made using a microcomputer, which through of browser was possible access the web page stored in the DE2 kit and send commands of control and monitoring to both TIMs (WTIM and STIM). The system describes a different form of development of the NCAP node to be applied in different environments with wired or wireless in the same node. © 2011 IEEE.

MATIUDE - monitoramento e acesso a transdutores inteligentes (Padrão IEEE 1451) utilizando dispositivos embarcados (celulares e PDA´s)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

MATIW 1451 - monitoramento e acionamento de transdutores inteligentes através da Web (Padrão IEEE 1451)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)