A Kinase-deficient Splice Variant of the Human JAK3 Is Expressed in Hematopoietic and Epithelial Cancer Cells

Signal transduction of cytokine receptors is mediated by the JAK family of tyrosine kinases. Recently, the kinase partners for the interleukin (IL)-2 receptor have been identified as JAK1 and JAK3. In this study, we report the identification of splice variants that may modulate JAK3 signaling. Three splice variants were isolated from different mRNA sources: breast (B), spleen (S), and activated monocytes (M). Sequence analysis revealed that the splice variants contain identical NH2-terminal regions but diverge at the COOH termini. Analyses of expression of the JAK3 splice isoforms by reverse transcriptase-polymerase chain reaction on a panel of cell lines show splice preferences in different cell lines: the S-form is more commonly seen in hematopoietic lines, whereas the B- and M-forms are detected in cells both of hematopoietic and epithelial origins. Antibodies raised against peptides to the B-form splice variant confirmed that the 125-kDa JAK3B protein product is found abundantly in hematopoietic as well as epithelial cells, including primary breast cancers. The lack of subdomain XI in the tyrosine kinase core of the B-form JAK3 protein suggests that it is a defective kinase. This is supported by the lack of detected autokinase activity of the B-form JAK3. Intriguingly, both the S and B splice isoforms of JAK3 appear to co-immunoprecipitate with the IL-2 receptor from HUT-78 cell lysates. This and the presence of multiple COOH-terminal splice variants coexpressed in the same cells suggest that the JAK3 splice isoforms are functional in JAK3 signaling and may enrich the complexity of the intracellular responses functional in IL-2 or cytokine signaling

The DACAPO-PESO campaign: Dynamics, Aerosol, Cloud and Precipitation Observations in the Pristine Environment of the Southern Ocean: An overview

This article gives an overview of the DACAPO-PESO field experiment, which has taken place in Punta Arenas, Chile, from November 2018 to November 2021, and showcases first exciting research results that have already emerged from it.In diesem Artikel wird ein Überblick über das DACAPO-PESO Experiment gegeben, welches von November 2018 bis November 2021 in Punta Arenas, Chile, stattgefunden hat. Außerdem werden erste spannende Forschungsergebnisse vorgestellt, die bereits daraus gewonnen wurden

Erythropoietin Receptor Signaling Is Membrane Raft Dependent

Upon erythropoietin (Epo) engagement, Epo-receptor (R) homodimerizes to activate JAK2 and Lyn, which phosphorylate STAT5. Although recent investigations have identified key negative regulators of Epo-R signaling, little is known about the role of membrane localization in controlling receptor signal fidelity. Here we show a critical role for membrane raft (MR) microdomains in creation of discrete signaling platforms essential for Epo-R signaling. Treatment of UT7 cells with Epo induced MR assembly and coalescence. Confocal microscopy showed that raft aggregates significantly increased after Epo stimulation (mean, 4.3±1.4(SE) vs. 25.6±3.2 aggregates/cell; p≤0.001), accompanied by a >3-fold increase in cluster size (p≤0.001). Raft fraction immunoblotting showed Epo-R translocation to MR after Epo stimulation and was confirmed by fluorescence microscopy in Epo stimulated UT7 cells and primary erythroid bursts. Receptor recruitment into MR was accompanied by incorporation of JAK2, Lyn, and STAT5 and their activated forms. Raft disruption by cholesterol depletion extinguished Epo induced Jak2, STAT5, Akt and MAPK phosphorylation in UT7 cells and erythroid progenitors. Furthermore, inhibition of the Rho GTPases Rac1 or RhoA blocked receptor recruitment into raft fractions, indicating a role for these GTPases in receptor trafficking. These data establish a critical role for MR in recruitment and assembly of Epo-R and signal intermediates into discrete membrane signaling units

Local erythropoietin and endothelial progenitor cells improve regional cardiac function in acute myocardial infarction

<p>Abstract</p> <p>Background</p> <p>Expanded endothelial progenitor cells (eEPC) improve global left ventricular function in experimental myocardial infarction (MI). Erythropoietin beta (EPO) applied together with eEPC may improve regional myocardial function even further by anti-apoptotic and cardioprotective effects. Aim of this study was to evaluate intramyocardial application of eEPCs and EPO as compared to eEPCs or EPO alone in experimental MI.</p> <p>Methods and Results</p> <p>In vitro experiments revealed that EPO dosed-dependently decreased eEPC and leukocyte apoptosis. Moreover, in the presence of EPO mRNA expression in eEPC of proangiogenic and proinflammatory mediators measured by TaqMan PCR was enhanced. Experimental MI was induced by ligation and reperfusion of the left anterior descending coronary artery of nude rats (n = 8-9). After myocardial transplantation of eEPC and EPO CD68+ leukocyte count and vessel density were enhanced in the border zone of the infarct area. Moreover, apoptosis of transplanted CD31 + TUNEL + eEPC was decreased as compared to transplantation of eEPCs alone. Regional wall motion of the left ventricle was measured using Magnetic Resonance Imaging. After injection of eEPC in the presence of EPO regional wall motion significantly improved as compared to injection of eEPCs or EPO alone.</p> <p>Conclusion</p> <p>Intramyocardial transplantation of eEPC in the presence of EPO during experimental MI improves regional wall motion. This was associated with an increased local inflammation, vasculogenesis and survival of the transplanted cells. Local application of EPO in addition to cell therapy may prove beneficial in myocardial remodeling.</p

Multiresolution analysis of the spatiotemporal variability in global radiation observed by a dense network of 99 pyranometers

The time series of global radiation observed by a dense network of 99 autonomous pyranometers during the HOPE campaign around Jülich, Germany, are investigated with a multiresolution analysis based on the maximum overlap discrete wavelet transform and the Haar wavelet. For different sky conditions, typical wavelet power spectra are calculated to quantify the timescale dependence of variability in global transmittance. Distinctly higher variability is observed at all frequencies in the power spectra of global transmittance under broken-cloud conditions compared to clear, cirrus, or overcast skies. The spatial autocorrelation function including its frequency dependence is determined to quantify the degree of similarity of two time series measurements as a function of their spatial separation. Distances ranging from 100 m to 10 km are considered, and a rapid decrease of the autocorrelation function is found with increasing frequency and distance. For frequencies above 1∕3 min−1 and points separated by more than 1 km, variations in transmittance become completely uncorrelated. A method is introduced to estimate the deviation between a point measurement and a spatially averaged value for a surrounding domain, which takes into account domain size and averaging period, and is used to explore the representativeness of a single pyranometer observation for its surrounding region. Two distinct mechanisms are identified, which limit the representativeness; on the one hand, spatial averaging reduces variability and thus modifies the shape of the power spectrum. On the other hand, the correlation of variations of the spatially averaged field and a point measurement decreases rapidly with increasing temporal frequency. For a grid box of 10 km  ×  10 km and averaging periods of 1.5–3 h, the deviation of global transmittance between a point measurement and an area-averaged value depends on the prevailing sky conditions: 2.8 (clear), 1.8 (cirrus), 1.5 (overcast), and 4.2 % (broken clouds). The solar global radiation observed at a single station is found to deviate from the spatial average by as much as 14–23 (clear), 8–26 (cirrus), 4–23 (overcast), and 31–79 W m−2 (broken clouds) from domain averages ranging from 1 km  ×  1 km to 10 km  ×  10 km in area