Secretome profiling of oral squamous cell carcinoma-associated fibroblasts reveals organization and disassembly of extracellular matrix and collagen metabolic process signatures

An important role has been attributed to cancer-associated fibroblasts (CAFs) in the tumorigenesis of oral squamous cell carcinoma (OSCC), the most common tumor of the oral cavity. Previous studies demonstrated that CAF-secreted molecules promote the proliferation and invasion of OSCC cells, inducing a more aggressive phenotype. In this study, we searched for differences in the secretome of CAFs and normal oral fibroblasts (NOF) using mass spectrometry-based proteomics and biological network analysis. Comparison of the secretome profiles revealed that upregulated proteins involved mainly in extracellular matrix organization and disassembly and collagen metabolism. Among the upregulated proteins were fibronectin type III domain-containing 1 (FNDC1), serpin peptidase inhibitor type 1 (SERPINE1), and stanniocalcin 2 (STC2), the upregulation of which was validated by quantitative PCR and ELISA in an independent set of CAF cell lines. The transition of transforming growth factor beta 1 (TGF-beta 1)-mediating NOFs into CAFs was accompanied by significant upregulation of FNDC1, SERPINE1, and STC2, confirming the participation of these proteins in the CAF-derived secretome. Type I collagen, the main constituent of the connective tissue, was also associated with several upregulated biological processes. The immunoexpression of type I collagen N-terminal propeptide (PINP) was significantly correlated in vivo with CAFs in the tumor front and was associated with significantly shortened survival of OSCC patients. Presence of CAFs in the tumor stroma was also an independent prognostic factor for OSCC disease-free survival. These results demonstrate the value of secretome profiling for evaluating the role of CAFs in the tumor microenvironment and identify potential novel therapeutic targets such as FNDC1, SERPINE1, and STC2. Furthermore, type I collagen expression by CAFs, represented by PINP levels, may be a prognostic marker of OSCC outcome.Peer reviewe

Use Of Computerized Manometry For The Detection Of Pharyngoesophageal Spasm In Tracheoesophageal Speech.

To evaluate the utility of computerized manometry (CM) to identify pharyngoesophageal segment (PES) spasm during tracheoesophageal speech. Prospective clinical, controlled study. Intraluminal pressures of the PES were collected in 12 tracheoesophageal speakers without spasm and 8 tracheoesophageal speakers with PES spasm before and after localized injection of botulinum toxin to the PES. All subjects underwent voice analysis and videofluoroscopy in addition to CM before and after treatment. All tracheoesophageal speakers with PES spasm presented with mean intraluminal pressures greater than 16 mmHg (mean, 25.36 mmHg). In contrast, mean intraluminal pressures of subjects without spasm was 11.76 mmHg (P<0.05). The negative predictive value associated with the use of 16 mmHg as a threshold value for spasm was 100%. CM is a clinically useful tool to aid in speech rehabilitation for tracheoesophageal speakers. Intraluminal pressures of greater than 16 mmHg was highly predictive for PES spasm.139449-5

Protein disulfide isomerase redox-dependent association with p47(phox): evidence for an organizer role in leukocyte NADPH oxidase activation

Mechanisms of leukocyte NADPH oxidase regulation remain actively investigated. We showed previously that vascular and macrophage oxidase complexes are regulated by the associated redox chaperone PDI. Here, we investigated the occurrence and possible underlying mechanisms of PDI-mediated regulation of neutrophil NADPH oxidase. In a semirecombinant cell-free system, PDI inhibitors scrRNase (100 mu g/mL) or bacitracin (1 mM) near totally suppressed superoxide generation. Exogenously incubated, oxidized PDI increased (by similar to 40%), whereas PDIred diminished (by similar to 60%) superoxide generation. No change occurred after incubation with PDI serine-mutated in all four redox cysteines. Moreover, a mimetic CxxC PDI inhibited superoxide production by similar to 70%. Thus, oxidized PDI supports, whereas reduced PDI down-regulates, intrinsic membrane NADPH oxidase complex activity. In whole neutrophils, immunoprecipitation and colocalization experiments demonstrated PDI association with membrane complex subunits and prominent thiol-mediated interaction with p47(phox) in the cytosol fraction. Upon PMA stimulation, PDI was mobilized from azurophilic granules to cytosol but did not further accumulate in membranes, contrarily to p47(phox). PDI-p47(phox) association in cytosol increased concomitantly to opposite redox switches of both proteins; there was marked reductive shift of cytosol PDI and maintainance of predominantly oxidized PDI in the membrane. Pulldown assays further indicated predominant association between PDIred and p47(phox) in cytosol. Incubation of purified PDI (> 80% reduced) and p47(phox) in vitro promoted their arachidonate-dependent association. Such PDI behavior is consistent with a novel cytosolic regulatory loop for oxidase complex (re) cycling. Altogether, PDI seems to exhibit a supportive effect on NADPH oxidase activity by acting as a redox-dependent enzyme complex organizer. J. Leukoc. Biol. 90: 799-810; 2011.Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq)Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior (CAPES

Resistência natural da madeira de Corymbia maculata (Hook.) K.D.Hill & L.A.S. Johnson a fungos e cupins xilófagos, em condições de laboratório Wood natural resistance of Corymbia maculata (Hook.) K.D.Hill & L.A.S Johnson to wood destroying fungi and termites, under laboratory tests

A pesquisa teve o objetivo de avaliar a resistência natural da madeira de Corymbia maculata a fungos e a cupins xilófagos, em condições de laboratório. De peças radiais (tábuas) que continham o cerne e o alburno intactos foram retirados corpos-de-prova de 2,00 x 2,00 x 1,00 cm, com a menor dimensão na direção tangencial (ensaio com fungos), e de 2,54 x 2,00 x 0,64 cm, com a maior dimensão na direção das fibras (ensaio com cupins), em quatro posições na direção medula-casca. As amostras foram submetidas à ação dos fungos Postia placenta, Neolentinus lepideus e Polyporus fumosus por 12 semanas, ou à ação de cupins do gênero Nasutitermes por 30 dias. Constatou-se que a resistência da madeira ao apodrecimento foi dependente da posição na direção medula-casca e dos fungos utilizados. As amostras retiradas nas posições mais externas do tronco foram mais deterioradas que as internas. Dentro de cada posição, os fungos causaram deterioração semelhante à madeira, exceto para a posição mais externa (alburno), em que o fungo P. fumosus causou menos deterioração que os demais. De modo geral, a madeira de C. maculata foi altamente resistente (posições internas) ou resistente (posições externas) aos fungos ensaiados. Somente para o fungo N. lepideus a posição mais externa foi moderadamente resistente. Quanto aos cupins, a resistência da madeira não foi afetada pela posição na direção medula-casca e apresentou uma baixa perda de massa para as posições analisadas. Além disto, os cupins causaram somente desgaste superficial à madeira, e morreram durante o ensaio, o que permitiu classificar a madeira de C.maculata como resistente aos cupins ensaiados.<br>This research evaluated the natural resistance of Corymbia maculata wood to wood-destroying fungi and termites, under laboratory tests. Radial pieces (boards), containing intact heartwood and sapwood were transformed into test samples measuring 2.00 x 2.00 x 1.00 cm, with the smallest measure in tangential direction (fungi assay) and measuring 2.45 x 2.00 x 0.64 cm; with the largerst measure fiberwise (termite assay), in four positions from pith to bark. The samples were submitted to fungi Postia placenta, Neolentinus lepideus and Polyporus fumosus action for 12 weeks or to Nasutitermes sp. action for 30 days. Wood decay resistance was found to be dependent on the position pith to bark and on the fungus. The wood samples in the most external positions were more deteriorated. Inside of each position, the fungi caused similar deterioration, except for the most external position (sapwood) where the P. fumosus caused less deterioration than other tested fungi. In general, C. maculata wood was highly resistant (internal positions) or resistant (external positions) to the fungi. Just for the Neolentinus lepideus, the external position was classified as moderately resistante. The tested wood to termite resistance was not affected by position in the pith to bark direction and showed weight loss for allanalyzed positions. Besides, it was affected only superficially and the termites died during the assay. The C. maculata wood was classified as resistant to the termites assayed

Confiabilidade do diagnóstico final de dengue na epidemia 2001-2002 no Município do Rio de Janeiro, Brasil Reliability of the final dengue diagnosis in the epidemic occurring in Rio de Janeiro, Brazil, 2001-2002

Este estudo analisou a confiabilidade do diagnóstico final das 155.242 notificações de dengue da epidemia 2001-2002 no Município do Rio de Janeiro, fornecidas pela Secretaria Municipal de Saúde do Rio de Janeiro, por meio do Sistema de Informação de Agravos de Notificação (SINAN). O diagnóstico final possui como opções de preenchimento: dengue clássico, febre hemorrágica do dengue, descartado, inconclusivo e ignorado. Foi construída uma rotina em Epi Info para comparar o diagnóstico final digitado no SINAN com os critérios do Ministério da Saúde (MS), agrupando os casos como dengue clássico, dengue hemorrágico, descartado e inconclusivo (inclui ignorado). O diagnóstico final mostrou 52,4% dengue clássico, 0,6% dengue hemorrágico, 0,9% descartado e 46% de inconclusivo e ignorado, sendo que 78% de dengue clássico, 69% de dengue hemorrágico e 21,1% dos descartados preencheram os critérios do MS. A confiabilidade do diagnóstico final digitado foi em geral satisfatória (kappa = 0,681; IC95%: 0,685-0,677), porém baixa para os óbitos (kappa = 0,152; IC95%: 0,046-0,258). Considerando-se o volume da epidemia, o diagnóstico final de dengue clássico e dengue hemorrágico foi satisfatório, porém a alta proporção de casos ignorados e inconclusivos e a baixa qualidade da informação nos óbitos limitam o uso do SINAN nesse contexto.<br>This study analyzed the reliability of the final diagnosis in the 155,242 dengue reports during the 2001-2002 epidemic in the city of Rio de Janeiro, Brazil, using the official information system on communicable diseases (SINAN). The system allows the following options for the final diagnosis: classic dengue, dengue hemorrhagic fever, discarded, inconclusive, and unknown. We built a classification routine in Epi Info to compare the final diagnosis from SINAN with Ministry of Health criteria. According to the final diagnosis, the case breakdown was: 52.4% classic dengue; 0.6% dengue hemorrhagic fever; 0.9% discarded; 46% inconclusive and unknown. The revised diagnosis showed that 78% of classic dengue, 69% of dengue hemorrhagic fever, and 21.1% of discarded cases met the classification criteria. Although the reliability of the SINAN final diagnosis was generally satisfactory (kappa = 0.681; 95%CI: 0.685-0.677), it was worse for fatal cases (kappa = 0.152; 95%CI: 0.046-0.258). Considering the epidemic's magnitude, the final diagnosis of classic dengue and dengue hemorrhagic fever was satisfactory, but the high proportion of inconclusive or unknown cases and the poor quality of information for fatal cases limit the usefulness of SINAN in this context

Panniculitis in childhood-onset systemic lupus erythematosus : a multicentric cohort study

To evaluate prevalence, clinical manifestations, laboratory abnormalities, treatment and outcome in a multicenter cohort of childhood-onset systemic lupus erythematosus (cSLE) patients with and without panniculitis. Panniculitis was diagnosed due to painful subcutaneous nodules and/or plaques in deep dermis/subcutaneous tissues and lobular/mixed panniculitis with lymphocytic lobular inflammatory infiltrate in skin biopsy. Statistical analysis was performed using Bonferroni correction(p < 0.004). Panniculitis was observed in 6/847(0.7%) cSLE. Painful subcutaneous erythematosus and indurated nodules were observed in 6/6 panniculitis patients and painful subcutaneous plaques in 4/6. Generalized distribution was evidenced in 3/6 and localized in upper limbs in 2/6 and face in 1/6. Cutaneous hyperpigmentation and/or cutaneous atrophy occurred in 5/6. Histopathology features showed lobular panniculitis without vasculitis in 5/6(one of them had concomitant obliterative vasculopathy due to antiphospholipid syndrome) and panniculitis with vasculitis in 1/6. Comparison between cSLE with panniculitis and 60 cSLE without panniculitis with same disease duration [2.75(0–11.4) vs. 2.83(0–11.8) years,p = 0.297], showed higher frequencies of constitutional involvement (67% vs. 10%,p = 0.003) and leukopenia (67% vs. 7%,p = 0.002). Cutaneous atrophy and hyperpigmentation occurred in 83% of patients. Panniculitis is a rare skin manifestation of cSLE occurring in the first three years of disease with considerable sequelae. The majority of patients have concomitant mild lupus manifestations591CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQ301805/2013-0; 303752/2015-7; 301479/2015-1; 305068/2014-8;303422/2015-7This study was supported by grants from Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq 301805/2013–0 to RMRP, 303752/2015–7 to MTT, 301479/2015–1 to CSM, 305068/2014–8 to EB and 303422/2015–7 to CAS), Federico Foundation (to EB, RMRP and CAS) and by Núcleo de Apoio à Pesquisa “Saúde da Criança e do Adolescente” da USP (NAP-CriAd) to CA