Bradykinin-induced Ca2+ signaling in human subcutaneous fibroblasts involves ATP release via hemichannels leading to P2Y12 receptors activation

Background: Chronic musculoskeletal pain involves connective tissue remodeling triggered by inflammatory mediators, such as bradykinin. Fibroblast cells signaling involve changes in intracellular Ca2+ ([Ca2+]i). ATP has been related to connective tissue mechanotransduction, remodeling and chronic inflammatory pain, via P2 purinoceptors activation. Here, we investigated the involvement of ATP in bradykinin-induced Ca2+ signals in human subcutaneous fibroblasts. Results: Bradykinin, via B2 receptors, caused an abrupt rise in [Ca2+]i to a peak that declined to a plateau, which concentration remained constant until washout. The plateau phase was absent in Ca2+-free medium; [Ca2+]i signal was substantially reduced after depleting intracellular Ca2+ stores with thapsigargin. Extracellular ATP inactivation with apyrase decreased the [Ca2+]i plateau. Human subcutaneous fibroblasts respond to bradykinin by releasing ATP via connexin and pannexin hemichannels, since blockade of connexins, with 2- octanol or carbenoxolone, and pannexin-1, with 10Panx, attenuated bradykinin-induced [Ca2+]i plateau, whereas inhibitors of vesicular exocytosis, such as brefeldin A and bafilomycin A1, were inactive. The kinetics of extracellular ATP catabolism favors ADP accumulation in human fibroblast cultures. Inhibition of ectonucleotidase activity and, thus, ADP formation from released ATP with POM-1 or by Mg2+ removal from media reduced bradykinin-induced [Ca2+]i plateau. Selective blockade of the ADP-sensitive P2Y12 receptor with AR-C66096 attenuated bradykinin [Ca2+]i plateau, whereas the P2Y1 and P2Y13 receptor antagonists, respectively MRS 2179 and MRS 2211, were inactive. Human fibroblasts exhibited immunoreactivity against connexin-43, pannexin-1 and P2Y12 receptor. Conclusions: Bradykinin induces ATP release from human subcutaneous fibroblasts via connexin and pannexin-1-containing hemichannels leading to [Ca2+]i mobilization through the cooperation of B2 and P2Y12 receptors.

Regulation of histone H2A.Z expression is mediated by sirtuin 1 in prostate cancer

Histone variants seem to play a major role in gene expression regulation. In prostate cancer, H2A.Z and its acetylated form are implicated in oncogenes’ upregulation. SIRT1, which may act either as tumor suppressor or oncogene, reduces H2A.Z levels in cardiomyocytes, via proteasome-mediated degradation, and this mechanism might be impaired in prostate cancer cells due to sirtuin 1 downregulation. Thus, we aimed to characterize the mechanisms underlying H2A.Z and SIRT1 deregulation in prostate carcinogenesis and how they interact. We found that H2AFZ and SIRT1 were up- and downregulated, respectively, at transcript level in primary prostate cancer and high-grade prostatic intraepithelial neoplasia compared to normal prostatic tissues. Induced SIRT1 overexpression in prostate cancer cell lines resulted in almost complete absence of H2A.Z. Inhibition of mTOR had a modest effect on H2A.Z levels, but proteasome inhibition prevented the marked reduction of H2A.Z due to sirtuin 1 overexpression. Prostate cancer cells exposed to epigenetic modifying drugs trichostatin A, alone or combined with 5-aza-2’-deoxycytidine, increased H2AFZ transcript, although with a concomitant decrease in protein levels. Conversely, SIRT1 transcript and protein levels increased after exposure. ChIP revealed an increase of activation marks within the TSS region for both genes. Remarkably, inhibition of sirtuin 1 with nicotinamide, increased H2A.Z levels, whereas activation of sirtuin 1 by resveratrol led to an abrupt decrease in H2A.Z. Finally, protein-ligation assay showed that exposure to epigenetic modifying drugs fostered the interaction between sirtuin 1 and H2A.Z. We concluded that sirtuin 1 and H2A.Z deregulation in prostate cancer are reciprocally related. Epigenetic mechanisms, mostly histone post-translational modifications, are likely involved and impair sirtuin 1-mediated downregulation of H2A.Z via proteasome-mediated degradation. Epigenetic modifying drugs in conjunction with enzymatic modulators are able to restore the normal functions of sirtuin 1 and might constitute relevant tools for targeted therapy of prostate cancer patients

Osteoporotic hip fractures: Bisphosphonates sales and observed turning point in trend. A population-based retrospective study

The aim is to examine the temporal trends of hip fracture incidence in Portugal by sex and age groups, and explore the relation with anti-osteoporotic medication. From the National Hospital Discharge Database, we selected from 1st January 2000 to 31st December 2008, 77,083 hospital admissions (77.4% women) caused by osteoporotic hip fractures (low energy, patients over 49 years-age), with diagnosis codes 820.x of ICD 9-CM. The 2001 Portuguese population was used as standard to calculate direct age-standardized incidence rates (ASIR) (100,000 inhabitants). Generalized additive and linear models were used to evaluate and quantify temporal trends of age specific rates (AR), by sex. We identified 2003 as a turning point in the trend of ASIR of hip fractures in women. After 2003, the ASIR in women decreased on average by 10.3 cases/100,000 inhabitants, 95% CI (− 15.7 to − 4.8), per 100,000 anti-osteoporotic medication packages sold. For women aged 65–69 and 75–79 we identified the same turning point. However, for women aged over 80, the year 2004 marked a change in the trend, from an increase to a decrease. Among the population aged 70–74 a linear decrease of incidence rate (95% CI) was observed in both sexes, higher for women: − 28.0% (− 36.2 to − 19.5) change vs − 18.8%, (− 32.6 to − 2.3). The abrupt turning point in the trend of ASIR of hip fractures in women is compatible with an intervention, such as a medication. The trends were different according to gender and age group, but compatible with the pattern of bisphosphonates sales.

O contacto no acolhimento familiar de crianças e jovens: Uma avaliação actual

No campo das múltiplas interações que ocorrem no acolhimento familiar, destaca-se a relação entre a criança acolhida e a família biológica, pelo impacto que tem no desenvolvimento da criança. Esta relação gera um conjunto de ligações que pode proporcionar ou dificultar à criança a possibilidade de partilhar valores, experiências e conselhos entre aqueles dois mundos. A manutenção das relações pessoais entre os pais e a criança acolhida é um direito de ambos, exceto se contrariar o interesse superior da criança, já que o corte abrupto das relações estabelecidas interfere na organização dos vínculos futuros. O presente artigo resulta da investigação em curso, no distrito do Porto, intitulado «Contacto no acolhimento familiar: padrões, resultados e modelos de gestão», no âmbito de atuação do InEd, o Centro de Investigação e Inovação em Educação da Escola Superior de Educação do Instituto Politécnico do Porto. Este estudo tem como objetivo analisar os resultados do contacto entre a criança em acolhimento familiar e a sua família de origem, bem como as causas da sua inexistência ou cessação. Os resultados preliminares evidenciam que a maioria das crianças e jovens têm contacto com a sua família de origem, ocorrendo em quase 50% dos casos na casa da família de acolhimento, mas nem sempre com a regularidade desejada. Apesar do impacto do contacto na criança ou jovem ser muito diversificado, a maioria manifesta reações emocionais e comportamentais positivas. Por fim, apresentam se implicações para a prática dos técnicos envolvidos neste fenómeno.