A reduction of the neutrophil migration into the site of infection during cecal ligation and puncture-induced sepsis increases host mortality. Inhibition of heme oxygenase (HO) prevents this ...neutrophil paralysis and improves host survival in the cecal ligation and puncture model. Taking into account that almost 50% of all sepsis cases are a consequence of pneumonia, we designed the present study to determine the role of HO in an experimental model of pneumonia-induced sepsis. The objective of this study was to evaluate whether the inhibition of HO improves the outcome and pathophysiologic changes of sepsis induced by an intratracheal instillation of Klebsiella pneumoniae. The pretreatment of mice subjected to pneumonia-induced sepsis with ZnDPBG (zinc deuteroporphyrin 2,4-bis glycol), a nonspecific HO inhibitor, increased the number of neutrophils in the bronchoalveolar spaces, reduced the bacterial load at the site of infection, and prevented the upregulation of CD11b and the downregulation of CXCR2 on blood neutrophils. Moreover, the pretreatment with ZnDPBG decreased alveolar collapse, attenuating the deleterious changes in pulmonary mechanics and gas exchanges and, as a consequence, improved the survival rate of mice from 0% to ∼20%. These results show that heme oxygenase is involved in the pathophysiology of pneumonia-induced sepsis and suggest that HO inhibitors could be helpful for the management of this disease.
Abstract
GPC3 was proposed as a negative regulator of cell proliferation in the mammary gland. Whereas normal breast tissues present high GPC3 levels, its expression is reduced in breast tumors. We ...have transfected LM3 murine mammary tumor cells with the GPC3 gene. We found that GPC3 reexpression inhibited metastasis in vivo. Although GPC3 signaling pathway is unclear, we reported that this glypican modulates Wnt, p38 and Akt pathways. However, regulation hierarchy has not been determined and we have no data in human cells. We believe it possible to generate an anti-metastasis therapy targeting GPC3 pathway. To test it, we developed a pre-clinical human model. First, we showed by qPCR that GPC3 expression levels are inversely proportional to the metastatic ability of a panel of human breast cancer cells. The metastatic Hs578T and MDA-MB231 cells expressed less GPC3 than the poorly-metastatic ZR-75-1 and MCF7 lines. We selected MDA-MB231 and MCF7 to be modified by genetic engineering. GPC3 expression was blocked by RNA interference technology. We transfected four shRNA-GPC3 and one shRNA scramble (shNC) into MCF7. After selection and cloning, we chose clone 2 that expressed about 75% less GPC3 for further characterization.
On the other hand, GPC3 was reexpressed in MDA-MB231 cells by transduction of pLV-GPC3-GFP (empty vector was used as control). GPC3 mRNA increased up to 200-fold in pLV-GPC3-GFP transduced cells. Then we evaluated cell properties in the generated models. Surprisingly, MDA-MB231-GPC3 cells showed a higher proliferation rate than controls, that became significant from 48 h of exponential growth (Population doubling time= 45 h for control vs. 24 h for GPC3 reexpressing cells). On the other side, MCF7-shGPC3 bulk and clone 2 proliferated faster than -shNC. This suggests that, in this cell line, GPC3 is inhibiting proliferation. GPC3 reexpression induced a 30% inhibition on MDA-MB231 viability when cells were serum starved for 96 h, while parental and -GFP remained 100% viable. No differences were detected among MCF7 engineered cells. So, the effect of GPC3 on cell viability would depend on the cellular context. We performed a wound healing assay to evaluate migration, determining that MDA-MB231-GPC3 cells were significantly less migrant than their controls (Wound coverage: MDA-MB231-GPC3 10% vs. parental and -GFP 90%). In sum, we have developed a human cell model to study the role of GPC3 on tumor progression. Our preliminary in vitro results reveal the importance of GPC3 in the biology of breast cancer cells.
Citation Format: Lilian F. Castillo, Rocio Tascon, María Amparo Lago Huvelle1, Ancély Ferreira dos Santos, Letícia Ferreira Terra, Rosangela A. m. Wailemann, Talita C. Oliveira, Mari Sogayar, Leticia Labriola, Elisa Bal de Kier Joffe, Maria Giselle Peters. Development of a human model to study the role of Glypican-3 (GPC3) on tumor progression of the mammary gland. abstract. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 508. doi:10.1158/1538-7445.AM2013-508
Hospital infections currently represent the interests of the international order, as it involves the performance of health professionals, the quality of physical facilities and materials of daily ...use. This study aimed to verify the recognition of the importance of hand hygiene in preventing nosocomial infection by health professionals. This is a cross-sectional study, which was applied in semi-structured form, a sample of 60 professionals, including nurses, technicians and / or licensed practical nurses and doctors, working in a public hospital located in Bahia, in October 2011, complying with ethical requirements for studies in humans. The results showed that 98.3% of respondents recognize the importance of hand hygiene in preventing hospital infection, 83.3% said they master the technique, however only 53.4% described it correctly. We conclude that despite the awareness of professionals about the importance and availability of products for hand hygiene, it is necessary to implement educational activities that motivate and intensify the adhesion of professionals
As infecções hospitalares representam atualmente uma preocupação de ordem internacional, pois envolve a atuação dos profissionais de saúde, a qualidade das instalações físicas e dos materiais de uso ...diário. Assim, considerando a relevância desse tema, esse estudo objetivou verificar o reconhecimento da importância da higienização das mãos na prevenção da infecção hospitalar pelos profissionais de saúde. Trata-se de um estudo transversal, em que foi aplicado formulário semiestruturado a uma amostra de 60 profissionais, entre enfermeiros, técnicos e /ou auxiliares de enfermagem e médicos, atuantes em um Hospital Público localizado na Bahia, em outubro de 2011, obedecendo às exigências éticas para estudos com seres humanos. Os resultados evidenciaram que 98,3% dos entrevistados reconhecem a importância da higienização das mãos na profilaxia à infecção hospitalar, 83,3% afirmaram dominar a técnica, entretanto, apenas 53,4% a descreveram corretamente. Conclui-se que apesar da conscientização dos profissionais sobre a importância e da disponibilidade dos produtos para higienização das mãos, é necessário a implementação de atividades pedagógicas que motivem e intensifiquem a adesão dos profissionais.
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