Chicken γδ T cells proliferate upon IL-2 and IL-12 treatment and show a restricted receptor repertoire in cell culture

In chickens, γδ T cells represent a large fraction of peripheral T cells; however, their function remains largely unknown. Here, we describe the selective in vitro expansion of γδ T cells from total splenocytes by stimulation with the cytokines IL-2 and IL-12. Under these conditions, γδ T cells proliferated preferentially and reached frequencies of >95% within three weeks. Although IL-2 alone also triggered proliferation, an increased proliferation rate was observed in combination with IL-12. Most of the expanded cells were γδ TCR and CD8 double-positive. Splenocytes sorted into TCR1+CD8+, TCR1highCD8−, and TCR1lowCD8− subsets proliferated well upon dual stimulation with IL-2/IL-12, indicating that none of the three γδ T cell subsets require bystander activation for proliferation. TCR1+CD8+ cells maintained CD8 surface expression during stimulation, whereas CD8− subpopulations showed varied levels of CD8 upregulation, with the highest upregulation observed in the TCR1high subset. Changes in the γδ T-cell receptor repertoire during cell culture from day 0 to day 21 were analyzed by next-generation sequencing of the γδ variable regions. Overall, long-term culture led to a restricted γ and δ chain repertoire, characterized by a reduced number of unique variable region clonotypes, and specific V genes were enriched at day 21. On day 0, the δ chain repertoire was highly diverse, and the predominant clonotypes differed between animals, while the most frequent γ-chain clonotypes were shared between animals. However, on day 21, the most frequent clonotypes in both the γ and δ chain repertoires were different between animals, indicating that selective expansion of dominant clonotypes during stimulation seems to be an individual outcome. In conclusion, IL-2 and IL-12 were sufficient to stimulate the in vitro outgrowth of γδ T cells. Analyses of the TCR repertoire indicate that the culture leads to an expansion of individual T cell clones, which may reflect previous in vivo activation. This system will be instrumental in studying γδ T cell function

The urban land use in the COSMO-CLM model: A comparison of three parameterizations for Berlin

The regional non-hydrostatic climate model COSMO-CLM is increasingly being used on fine spatial scales of 1–5 km. Such applications require a detailed differentiation between the parameterization for natural and urban land uses. Since 2010, three parameterizations for urban land use have been incorporated into COSMO-CLM. These parameterizations vary in their complexity, required city parameters and their computational cost. We perform model simulations with the COSMO-CLM coupled to these three parameterizations for urban land in the same model domain of Berlin on a 1-km grid and compare results with available temperature observations. While all models capture the urban heat island, they differ in spatial detail, magnitude and the diurnal variation

1st ESMO Consensus Conference in lung cancer; Lugano 2010: Small-cell lung cancer

The 1st ESMO Consensus Conference on lung cancer was held in Lugano, Switzerland on 21st and 22nd May 2010 with the participation of a multidisciplinary panel of leading professionals in pathology and molecular diagnostics and medical, surgical and radiation oncology. Before the conference, the expert panel prepared clinically relevant questions concerning five areas as follows: early and locally advanced non-small-cell lung cancer (NSCLC), first-line metastatic NSCLC, second-/third-line NSCLC, NSCLC pathology and molecular testing, and small-cell lung cancer (SCLC) to be addressed through discussion at the Consensus Conference. All relevant scientific literature for each question was reviewed in advance. During the Consensus Conference, the panel developed recommendations for each specific question. The consensus agreement in SCLC is reported in this article. The recommendations detailed here are based on an expert consensus after careful review of published data. All participants have approved this final updat

Interval-Based Adaptive Inertia and Damping Control of a Virtual Synchronous Machine

This paper presents a novel virtual synchronous machine controller for converters in power systems with a high share of renewable resources. Using an interval-based approach, the emulated inertia and damping constants are adaptively adjusted according to the frequency disturbance in the system, while simultaneously keeping the frequency within prescribed limits. Furthermore, the sufficient stability conditions for control tuning are derived. The proposed design is integrated into a state-of-the-art converter control scheme and tested through time-domain simulations. A comparative study against the existing approaches in the literature verifies the control effectiveness

Group suckling in organic sow units

Group suckling - a combined system of single and group housing of lactating sows - appears a suitable system for organic pig production. The aim of the study was to describe the status quo of group suckling in organic farms. 31 organic sow units in Germany, Austria and Switzerland were investigated. Stockmen were interviewed, stables were inspected and animals were examined during three visits on each farm enterprise, respectively. The majority of farms kept three sows with piglets in one group suckling unit. 76 % of the group suckling sows (n=192) were in a good nutritional condition, 18 % were considered thin and 8 % of sows were too fat. Relatively few sows showed skin lesions caused by poor housing conditions. Only 18 of 203 sows behaved anxiously or aggressively. On average 9.1 piglets per sow and litter were weaned. Amongst the investigated farms, none was optimally managed. However, no plausible correlations between biological performance, animal health, human-animal relationship on the one hand and farm-specific production conditions (housing, management, feeding, watering) on the other hand were determined. It can therefore be deduced that the “success” or “failure” of the study farms can be attributed to the interaction of different factors rather than to individual production factors

Long-term benefit of lurbinectedin as palliative chemotherapy in progressive malignant pleural mesothelioma (MPM): final efficacy and translational data of the SAKK 17/16 study.

BACKGROUND The SAKK 17/16 study showed promising efficacy data with lurbinectedin as second- or third-line palliative therapy in malignant pleural mesothelioma. Here, we evaluated long-term outcome and analyzed the impact of lurbinectedin monotherapy on the tumor microenvironment at the cellular and molecular level to predict outcomes. MATERIAL AND METHODS Forty-two patients were treated with lurbinectedin in this single-arm study. Twenty-nine samples were available at baseline, and seven additional matched samples at day one of cycle two of treatment. Survival curves and rates between groups were compared using the log-rank test and Kaplan-Meier method. Statistical significance was set at P value <0.05. RESULTS Updated median overall survival (OS) was slightly increased to 11.5 months [95% confidence interval (CI) 8.8-13.8 months]. Thirty-six patients (85%) had died. The OS rate at 12 and 18 months was 47% (95% CI 32.1% to 61.6%) and 31% (95% CI 17.8% to 45.0%), respectively. Median progression-free survival was 4.1 months (95% CI 2.6-5.5 months). No new safety signals were observed. Patients with lower frequencies of regulatory T cells, as well as lower tumor-associated macrophages (TAMs) at baseline, had a better OS. Comparing matched biopsies, a decrease of M2 macrophages was observed in five out of seven patients after exposure to lurbinectedin, and two out of four patients showed increased CD8+ T-cell infiltrates in tumor. DISCUSSION Lurbinectedin continues to be active in patients with progressing malignant pleural mesothelioma. According to our very small sample size, we hypothesize that baseline TAMs and regulatory T cells are associated with survival. Lurbinectedin seems to inhibit conversion of TAMs to M2 phenotype in humans

Unraveling the sequence of serpentinization reactions : petrography, mineral chemistry, and petrophysics of serpentinites from MAR 15°N (ODP Leg 209, Site 1274)

Author Posting. © American Geophysical Union, 2006. This article is posted here by permission of American Geophysical Union for personal use, not for redistribution. The definitive version was published in Geophysical Research Letters 33 (2006): L13306, doi:10.1029/2006GL025681.The results of detailed textural, mineral chemical, and petrophysical studies shed new light on the poorly constrained fluid-rock reaction pathways during retrograde serpentinization at mid-ocean ridges. Uniformly depleted harzburgites and dunites from the Mid-Atlantic Ridge at 15°N show variable extents of static serpentinization. They reveal a simple sequence of reactions: serpentinization of olivine and development of a typical mesh texture with serpentine-brucite mesh rims, followed by replacement of olivine mesh centers by serpentine and brucite. The serpentine mesh rims on relic olivine are devoid of magnetite. Conversely, domains in the rock that are completely serpentinized show abundant magnetite. We propose that low-fluid-flux serpentinization of olivine to serpentine and ferroan brucite is followed by later stages of serpentinization under more open-system conditions and formation of magnetite by the breakdown of ferroan brucite. Modeling of this sequence of reactions can account for covariations in magnetic susceptibility and grain density of the rocks.Funding for this research was provided by USSSP and NSF-OCE grant 9986135. WB acknowledges support through a fellowship by the Deep Ocean Exploration Institute

The Epstein-Barr Virus G-Protein-Coupled Receptor Contributes to Immune Evasion by Targeting MHC Class I Molecules for Degradation

Epstein-Barr virus (EBV) is a human herpesvirus that persists as a largely subclinical infection in the vast majority of adults worldwide. Recent evidence indicates that an important component of the persistence strategy involves active interference with the MHC class I antigen processing pathway during the lytic replication cycle. We have now identified a novel role for the lytic cycle gene, BILF1, which encodes a glycoprotein with the properties of a constitutive signaling G-protein-coupled receptor (GPCR). BILF1 reduced the levels of MHC class I at the cell surface and inhibited CD8+ T cell recognition of endogenous target antigens. The underlying mechanism involves physical association of BILF1 with MHC class I molecules, an increased turnover from the cell surface, and enhanced degradation via lysosomal proteases. The BILF1 protein of the closely related CeHV15 c1-herpesvirus of the Rhesus Old World primate (80% amino acid sequence identity) downregulated surface MHC class I similarly to EBV BILF1. Amongst the human herpesviruses, the GPCR encoded by the ORF74 of the KSHV c2-herpesvirus is most closely related to EBV BILF1 (15% amino acid sequence identity) but did not affect levels of surface MHC class I. An engineered mutant of BILF1 that was unable to activate G protein signaling pathways retained the ability to downregulate MHC class I, indicating that the immune-modulating and GPCR-signaling properties are two distinct functions of BILF1. These findings extend our understanding of the normal biology of an important human pathogen. The discovery of a third EBV lytic cycle gene that cooperates to interfere with MHC class I antigen processing underscores the importance of the need for EBV to be able to evade CD8+ T cell responses during the lytic replication cycle, at a time when such a large number of potential viral targets are expressed