Acid-sensing ion channel 3 decreases phosphorylation of extracellular signal-regulated kinases and induces synoviocyte cell death by increasing intracellular calcium.

IntroductionAcid-sensing ion channel 3 (ASIC3) is expressed in synoviocytes, activated by decreases in pH, and reduces inflammation in animal models of inflammatory arthritis. The purpose of the current study was to characterize potential mechanisms underlying the control of inflammation by ASIC3 in fibroblast-like synoviocytes (FLS).MethodsExperiments were performed in cultured FLS from wild-type (WT) and ASIC3-/- mice, ASIC1-/- mice, and people with rheumatoid arthritis. We assessed the effects of acidic pH with and without interleukin-1β on FLS and the role of ASICs in modulating intracellular calcium [Ca(2+)](i), mitogen activated kinase (MAP kinase) expression, and cell death. [Ca(2+)](i) was assessed by fluorescent calcium imaging, MAP kinases were measured by Western Blots; ASIC, cytokine and protease mRNA expression were measured by quantitative PCR and cell death was measured with a LIVE/DEAD assay.ResultsAcidic pH increased [Ca(2+)](i) and decreased p-ERK expression in WT FLS; these effects were significantly smaller in ASIC3-/- FLS and were prevented by blockade of [Ca(2+)]i. Blockade of protein phosphatase 2A (PP2A) prevented the pH-induced decreases in p-ERK. In WT FLS, IL-1β increases ASIC3 mRNA, and when combined with acidic pH enhances [Ca(2+)](i), p-ERK, IL-6 and metalloprotienase mRNA, and cell death. Inhibitors of [Ca(2+)](i) and ERK prevented cell death induced by pH 6.0 in combination with IL-1β in WT FLS.ConclusionsDecreased pH activates ASIC3 resulting in increased [Ca(2+)](i), and decreased p-ERK. Under inflammatory conditions, acidic pH results in enhanced [Ca(2+)](i) and phosphorylation of extracellular signal-regulated kinase that leads to cell death. Thus, activation of ASIC3 on FLS by acidic pH from an inflamed joint could limit synovial proliferation resulting in reduced accumulation of inflammatory mediators and subsequent joint damage

Parasitismo de larvas da mosca-do-mediterrâneo por Diachasmimorpha longicaudata (Ashmead) (Hymenoptera: Braconidae) em diferentes cultivares de goiaba.

As moscas-das-frutas são as pragas que causam os maiores prejuízos à fruticultura mundial. Esses prejuízos podem ser diretos, com perdas na produção e indiretos. Por serem pragas quarentenárias, existem barreiras comerciais impostas pelos países importadores, limitando a exportação de frutos in natura. Apesar de o Brasil ser o terceiro maior produtor mundial de frutas, exporta-se apenas 2% dessa produção, enquanto que Diachasmimorpha longicaudata infestadas com larvas de Ceratitis capitata (Wiedemann) (Diptera: Tephritidae). Cinco lotes de oito frutos de goiaba, sendo dois frutos por cultivar, foram acondicionados em gaiolas contendo adultos de C. capitata, por 2h para oviposição. Após uma semana, quando as larvas já haviam se desenvolvido dentro dos frutos, estes foram expostos aos parasitóides durante 24h. Foram avaliados o peso médio dos frutos, a mortalidade das larvas, o número de pupários e as porcentagens de moscas e parasitóides emergidos

The Dynamics of Radiative Shock Waves: Linear and Nonlinear Evolution

The stability properties of one-dimensional radiative shocks with a power-law cooling function of the form $\Lambda \propto \rho^2T^\alpha$ are the main subject of this work. The linear analysis originally presented by Chevalier & Imamura, is thoroughfully reviewed for several values of the cooling index $\alpha$ and higher overtone modes. Consistently with previous results, it is shown that the spectrum of the linear operator consists in a series of modes with increasing oscillation frequency. For each mode a critical value of the cooling index, $\alpha_\textrm{c}$, can be defined so that modes with $\alpha < \alpha_\textrm{c}$ are unstable, while modes with $\alpha > \alpha_\textrm{c}$ are stable. The perturbative analysis is complemented by several numerical simulations to follow the time-dependent evolution of the system for different values of $\alpha$. Particular attention is given to the comparison between numerical and analytical results (during the early phases of the evolution) and to the role played by different boundary conditions. It is shown that an appropriate treatment of the lower boundary yields results that closely follow the predicted linear behavior. During the nonlinear regime, the shock oscillations saturate at a finite amplitude and tend to a quasi-periodic cycle. The modes of oscillations during this phase do not necessarily coincide with those predicted by linear theory, but may be accounted for by mode-mode coupling.Comment: 33 pages, 12 figures, accepted for publication on the Astrophysical Journa

Improved Classification Using Hidden Markov Averaging From Multiple Observation Sequences

The enormous popularity of Hidden Markov models (HMMs) in spatio-temporal pattern recognition is largely due to the ability to 'learn' model parameters from observation sequences through the Baum-Welch and other re-estimation procedures. In this study, HMM parameters are estimated from an ensemble of models trained on individual observation sequences. The proposed methods are shown to provide superior classification performance to competing methods

ORP3 splice variants and their expression in human tissues and hematopoietic cells

ORP3 is a member of the newly described family of oxysterol-binding protein (OSBP)-related proteins (ORPs). We previously demonstrated that this gene is highly expressed in CD34+ hematopoietic progenitor cells, and deduced that the &quot;full-length&quot; ORP3 gene comprises 23 exons and encodes a predicted protein of 887 amino acids with a C-terminal OSBP domain and an N-terminal pleckstrin homology domain. To further characterize the gene, we cloned ORP3 cDNA from PCR products and identified multiple splice variants. A total of eight isoforms were demonstrated with alternative splicing of exons 9, 12, and 15. Isoforms with an extension to exon 15 truncate the OSBP domain of the predicted protein sequence. In human tissues there was specific isoform distribution, with most tissues expressing varied levels of isoforms with the complete OSBP domain; while only whole brain, kidney, spleen, thymus, and thyroid expressed high levels of the isoforms associated with the truncated OSBP domain. Interestingly, the expression in cerebellum, heart, and liver of most isoforms was negligible. These data suggest that differential mRNA splicing may have resulted in functionally distinct forms of the ORP3 gene.<br /

Benzoyl­dicarbon­yl(η5-inden­yl)ruthenium(II)

In the title mol­ecule, [Ru(C9H7)(C7H5O)(CO)2], the dihedral angle between the mean plane of the indene ring system and the phenyl ring is 86.28 (8)°. The crystal structure is stabilized by weak inter­molecular C—H⋯O and C—H⋯π(arene) inter­actions. The Ru—η5-cyclopentadienyl centroid bond length is 1.946 (11) 

RTKN2 induces NF-KappaB dependent resistance to intrinsic apoptosis in HEK cells and REgulates BCL-2 genes in human CD4+ lymphocytes

The gene for Rhotekin 2 (RTKN2) was originally identified in a promyelocytic cell line resistant to oxysterol-induced apoptosis. It is differentially expressed in freshly isolated CD4+ T-cells compared with other hematopoietic cells and is down-regulated following activation of the T-cell receptor. However, very little is known about the function of RTKN2 other than its homology to Rho-GTPase effector, rhotekin, and the possibility that they may have similar roles. Here we show that stable expression of RTKN2 in HEK cells enhanced survival in response to intrinsic apoptotic agents; 25-hydroxy cholesterol and camptothecin, but not the extrinsic agent, TNF&alpha;. Inhibitors of NF-KappaB, but not MAPK, reversed the resistance and mitochondrial pro-apoptotic genes, Bax and Bim, were down regulated. In these cells, there was no evidence of RTKN2 binding to the GTPases, RhoA or Rac2. Consistent with the role of RTKN2 in HEK over-expressing cells, suppression of RTKN2 in primary human CD4+ T-cells reduced viability and increased sensitivity to 25-OHC. The expression of the pro-apoptotic genes, Bax and Bim were increased while BCL-2 was decreased. In both cell models RTKN2 played a role in the process of intrinsic apoptosis and this was dependent on either NF-KappaB signaling or expression of downstream BCL-2 genes. As RTKN2 is a highly expressed in CD4+ T-cells it may play a role as a key signaling switch for regulation of genes involved in T-cell survival.<br /