Effect of cytoglobin overexpression on extracellular matrix component synthesis in human tenon fibroblasts

BACKGROUND: Conjunctival filtering bleb scar formation is the main reason for the failure of glaucoma filtration surgery. Cytoglobin (Cygb) has been reported to play an important role in extracellular matrix (ECM) remodeling, fibrosis and tissue damage repairing. This study aimed to investigate the role of Cygb in anti-scarring during excessive conjunctival wound healing after glaucoma filtration surgery. METHODS: Cygb was overexpressed in human tenon fibroblasts (hTFs) by transfecting hTFs with lentiviral particles encoding pLenti6.2-FLAG-Cygb. Changes in the mRNA and protein levels of fibronectin, collagen I, collagen III, TGF-ß1, and HIF1α were determined by RT-PCR and western blotting respectively. RESULTS: After Cygb overexpression, hTFs displayed no significant changes in visual appearance and cell counts compared to controls. Whereas, Cygb overexpression significantly decreased the mRNA and protein expression levels of collagen I, collagen III and fibronectin compared with control (p < 0.01). There was also a statistically significant decrease in the mRNA and protein levels of TGF-ß1 and HIF-1α in hTFs with overexpressed Cygb compared with control group (p < 0.05). CONCLUSION: Our study provided evidence that overexpression of Cygb decreased the expression levels of fibronectin, collagen I, collagen III, TGF-ß1 and HIF-1α in hTFs. Therefore, therapies targeting Cygb expression in hTFs may pave a new way for clinicians to solve the problem of post-glaucoma surgery scarring.

Risk assessment for preterm delivery using the fetal fibronectin test associated with the measurement of uterine cervix length in symptomatic pregnant women / Avaliação do risco para parto pré-termo utilizando o teste da fibronectina fetal, associado à medida de comprimento de colo uterino, em gestantes sintomáticas

Abstract Objective To analyze the use of the measurement of uterine cervix length (MUCL) and the fetal fibronectin (fFN) rapid test as predictors of preterm delivery (PTD) in symptomatic pregnant women assisted at the Santa Casa de Misericórdia de Sobral Maternity Hospital. Methods This was a prospective and analytic study involving 53 parturients assisted between September of 2015 and July of 2016; the participants were between 24 and 34 weeks of gestational age (GA) and presented complaints related to preterm labor (PTL) prodromes. Vaginal secretion was collected for fFN testing, and the MUCL was obtained via transvaginal ultrasonography. Results A total of 58.49% of the subjects showed MUCL < 25 mm, and 41.51% were positive in the fFNrapid test.Atotal of 48 patients were followed-up until their delivery date, and 54.17% resulted in PTL. The relative risk (RR) for PTD in patients with MUCL < 25 mm was 1.83 (p = 0.09, 0.99-3.36, 95% confidence interval [CI]), with a mean time before delivery of 2.98 weeks. Based on fFN positive results, the RR was 3.50 (p = 0.002, 1.39- 8.79, 95%CI) and themean time until delivery was 1.94weeks. The RRwas 2.70 (p = 0.002, 1.08-6.72, 95%CI) when both tests were used. The RR of PTD within 48 hours, and 7 and 14 days were, respectively, 1.30 (p = 0.11, 95% CI 1.02-1.67), 1.43 (p = 0.12, 95% CI % 0.99-2.06), and 2.03 (p = 0.008, 95% CI 1.26-3.27), when based on the MUCL, and 1.75 (p = 0.0006, 95% CI 1.20-2.53), 2.88 (p = 0.0001, 95% CI, 1.57-5.31), and 3.57 (p = 0.0002, 95% CI 1.63-7.81) when based on positive fFN results. The RR at 48 hours and 7 and 14 days considering both tests was 1.74 (p = 0.0001, 95% CI 1.14-2.64), 2.22 (p = 0.0001, 95% CI 1.22-4.04), and 2.76 (p = 0.0002, 95% CI 1.27-5.96), respectively. Conclusion In symptomatic pregnant women, we concluded that the MUCL < 25 mm associated with positive fFN rapid test indicate increased the risk for PTD. Further studies with larger sample sizes could contribute in supporting the results presented in the current study.

Resumo Objetivo Analisar a utilização da medida do comprimento do colo uterino (MCCU), e do teste da fibronectina fetal (FNf) como preditores do trabalho de parto pré-termo (PPT), em gestantes sintomáticas, atendidas na Maternidade da Santa Casa de Misericórdia de Sobral. Métodos Foi realizado umestudo prospectivo e analítico, envolvendo 53 parturientes atendidas no período de setembro de 2015 a julho de 2016, com idade gestacional (IG) entre 24 e 34 semanas que tiveram queixas relacionadas a pródromos de trabalho de parto prematuro (TPP), sendo realizada coleta de secreção vaginal para FNf e MCCU por via ultrassonográfica transvaginal. Resultados Um total de 58,49% das pacientes tinham MCCU < 25 mm, e 41,51% tiveram teste rápido de fFN positivo. Foi feito o acompanhamento de 48 pacientes, com 54,17% de PPTs. O risco relativo (RR) para PPT com MCCU < 25 mm foi de 1,83 (p = 0,09, 0,99-3,36, intervalo de confiança [IC] 95%), com média de tempo até o parto de 2,98 semanas. Para fFN, o RR foi de 3.50 (p = 0.002, 1.39-8.79, IC 95%) e a média até o parto foi de 1,94 semanas. Quando os dois testes forampositivos, o RR foi de 2,70 (1,08-6,72). Para a MCCU, o RR para PPT em 48 horas, 7 e 14 dias foram 1,30 (p = 0.11, 95% IC 1.02-1.67), 1,43 (p = 0.12, 95% CI % 0.99-2.06) e 2,03 (p = 0.008, 95% IC 1.26-3.27), respectivamente. Para FNf, em 48 horas, 7 e 14 dias foi de 1,75 (p = 0.0006, 95% IC 1.20-2.53, 2,88 (p = 0.0001, 95% IC, 1.57-5.31) e 3,57 (p = 0.0002, 95% IC 1.63-7.81) respectivamente. Com os dois testes, o RR em 48 horas, 7 e 14 dias foi 1,74 (p = 0.0001, 95%IC 1.14-2.64), 2,22 (p = 0.0001, 95% IC 1.22-4.04) e 2,76 (p = 0.0002, 95% IC 1.27-5.96) respectivamente. Conclusão Em mulheres grávidas sintomáticas, concluímos que a MCCU < 25 mm e o teste rápido de FNf positivo indicam aumento do risco de PPT. Outros estudos com tamanhos de amostra maiores podem contribuir para apoiar os resultados apresentados no presente estudo.

Decellularization of placentas: establishing a protocol

Biological biomaterials for tissue engineering purposes can be produced through tissue and/or organ decellularization. The remaining extracellular matrix (ECM) must be acellular and preserve its proteins and physical features. Placentas are organs of great interest because they are discarded after birth and present large amounts of ECM. Protocols for decellularization are tissue-specific and have not been established for canine placentas yet. This study aimed at analyzing a favorable method for decellularization of maternal and fetal portions of canine placentas. Canine placentas were subjected to ten preliminary tests to analyze the efficacy of parameters such as the type of detergents, freezing temperatures and perfusion. Two protocols were chosen for further analyses using histology, scanning electron microscopy, immunofluorescence and DNA quantification. Sodium dodecyl sulfate (SDS) was the most effective detergent for cell removal. Freezing placentas before decellularization required longer periods of incubation in different detergents. Both perfusion and immersion methods were capable of removing cells. Placentas decellularized using Protocol I (1% SDS, 5 mM EDTA, 50 mM TRIS, and 0.5% antibiotic) preserved the ECM structure better, but Protocol I was less efficient to remove cells and DNA content from the ECM than Protocol II (1% SDS, 5 mM EDTA, 0.05% trypsin, and 0.5% antibiotic).

Cells involved in extracellular matrix remodeling after acute myocardial infarction / Células envolvidas no remodelamento da matriz extracelular após infarto agudo do miocárdio

Objective Evaluate the effects of VEGF165 gene transfer in the process of remodeling of the extracellular matrix after an acute myocardial infarct. Methods Wistar rats were submitted to myocardial infarction, after the ligation of the left descending artery, and the left ventricle ejection fraction was used to classify the infarcts into large and small. The animals were divided into groups of ten, according to the size of infarcted area (large or small), and received or not VEGF165 treatment. Evaluation of different markers was performed using immunohistochemistry and digital quantification. The primary antibodies used in the analysis were anti-fibronectin, anti-vimentin, anti-CD44, anti-E-cadherin, anti-CD24, anti-alpha-1-actin, and anti-PCNA. The results were expressed as mean and standard error, and analyzed by ANOVA, considering statistically significant if p≤0.05. Results There was a significant increase in the expression of undifferentiated cell markers, such as fibronectin (protein present in the extracellular matrix) and CD44 (glycoprotein present in the endothelial cells). However, there was decreased expression of vimentin and PCNA, indicating a possible decrease in the process of cell proliferation after treatment with VEGF165. Markers of differentiated cells, E-cadherin (adhesion protein between myocardial cells), CD24 (protein present in the blood vessels), and alpha-1-actin (specific myocyte marker), showed higher expression in the groups submitted to gene therapy, compared to non-treated group. The value obtained by the relation between alpha-1-actin and vimentin was approximately three times higher in the groups treated with VEGF165, suggesting greater tissue differentiation. Conclusion The results demonstrated the important role of myocytes in the process of tissue remodeling, confirming that VEGF165 seems to provide a protective effect in the treatment of acute myocardial infarct. .

Objetivo Avaliar os efeitos da transferência gênica do VEGF165 no processo de remodelamento da matriz extracelular após infarto agudo do miocárdio. Métodos Ratos Wistar foram submetidos ao infarto do miocárdio por ligação da artéria coronária descendente esquerda, e a fração de ejeção de ventrículo esquerdo foi utilizada para classificar os infartos em grandes e pequenos. Os animais foram divididos em grupos de dez animais, de acordo com o tamanho do infarto (grande ou pequeno), e receberam ou não tratamento com o VEGF165. A avaliação dos diferentes marcadores foi realizada por imuno-histoquímica e quantificação digital. Os anticorpos primários utilizados foram antifibronectina, antivimentina, anti- CD44, anti-E-caderina, anti-CD24, anti-alfa-1-actina e anti-PCNA. Os resultados foram representados como média e erro padrão, e analisados por ANOVA, sendo considerado estatisticamente significativo se p≤0,05. Resultados Houve aumento significativo da expressão de marcadores de células indiferenciadas, como fibronectina (proteína presente na matriz extracelular) e CD44 (glicoproteína presente nas células endoteliais). Entretanto, houve diminuição da expressão de vimentina e PCNA, indicando possível diminuição do processo de proliferação celular após o tratamento com VEGF165. Os marcadores de células diferenciadas, E-caderina (proteína de adesão entre as células do miocárdio), CD24 (proteína presente nos vasos sanguíneos) e alfa-1-actina (marcador especifico de miócitos) também apresentaram maior expressão nos grupos submetidos à terapia gênica, comparativamente com o grupo não tratado. O valor obtido pela relação entre alfa-1-actina e vimentina foi aproximadamente três vezes maior nos grupos tratados com VEGF165, indicando maior diferenciação tecidual. Conclusão O papel dos miócitos se mostrou importante no processo de remodelamento tecidual, confirmando que o VEGF165 parece conferir um efeito protetor no tratamento do infarto agudo do miocárdio. .

Tenascin and Fibronectin Expression after Pulp Capping with Different Hemostatic Agents: A Preliminary Study

This study investigated the expression of extracellular matrix glycoproteins tenascin (TN) and fibronectin (FN) in pulp repair after capping with calcium hydroxide (CH), following different hemostasis protocols. Class I cavities with a pulp exposure were prepared in 42 human third molars scheduled for extraction. Different hemostatic agents (0.9% saline solution, 5.25% sodium hypochlorite and 2% chlorhexidine digluconate) were used and pulps were capped with CH cement. After 7, 30 or 90 days, teeth were extracted, formalin-fixed, and prepared for immunohistochemical technique. Hemostatic agents did not influence the expression of TN and FN. Both glycoproteins were found in the entire the pulp tissue and around collagen fibers, but were absent in the mineralized tissues. In the predentin, TN showed positive immunostaining and FN had a variable expression. Within 7 days post-treatment, a slightly more pronounced immunostaining on the pulp exposure site was observed. Within 30 days, TN and FN demonstrated a positive expression around the dentin barrier and at 90 days, a thin and linear expression of TN and FN was delimitating the reparative dentin. In conclusion, hemostatic agents did not influence TN and FN expression. Immunostaining for TN and FN was seen in different regions and periods, demonstrating their role in pulp repair.

Este estudo investigou a expressão das glicoproteínas Tenascina (TN) e Fibronectina (FN) da matriz extracelular no reparo pulpar após capeamento com hidróxido de cálcio (HC), seguindo diferentes protocolos de hemostasia. Cavidades de classe I com exposição pulpar foram preparadas em 42 terceiros molares humanos indicados para extração. Diferentes agentes hemostáticos (solução salina a 0,9%, hipoclorito de sódio a 5,25% e clorexidina a 2%) foram usados e as polpas foram capeadas com cimento de HC. Após 7, 30 ou 90 dias, os dentes foram extraídos, fixados em formalina e preparados para análise imunoistoquímica. Os agentes hemostáticos não influenciaram a expressão de TN e FN. Ambas glicoproteínas foram encontradas em todo tecido pulpar, ao redor das fibras colágenas e estiveram ausentes nos tecidos mineralizados. Na pré-dentina, a TN mostrou forte imunoexpressão e a FN teve uma expressão variável. Após 7 dias, foi observada uma expressão levemente mais pronunciada no lugar da exposição pulpar. Aos 30 dias, a TN e a FN demonstraram uma expressão mais forte sob a barreira dentinária e aos 90 dias, uma expressão fina e linear da TN e FN apresentava-se delimitando a dentina reparativa. Em conclusão, os agentes hemostáticos não influenciaram e expressão da TN e da FN. A imunoexpressão da TN e FN foi observada em diferentes regiões e períodos, demonstrando o seu papel no reparo pulpar.

Imatinib atenua a fibrose miocárdica em associação com a inibição da atividade do PDGFRα / Imatinib attenuates myocardial fibrosis in association with inhibition of the PDGFRα activity

FUNDAMENTO: O Imatinib é um inibidor do receptor tirosina-quinase que foi confirmada como exercendo um efeito inibidor sobre a atividade do receptor do PDGF, fator de crescimento plaquetário (PDGFRα e PDGFRβ). OBJETIVO: Investigar o efeito protetor do Imatinib na fibrose miocárdica em acetato de deoxicorticosterona (DOCA)/ratos com hipertensão induzida por sal. MÉTODOS: Sessenta ratos Sprague-Dawley machos, uninefrectomizados foram distribuídos em três grupos: ratos controles (grupo CON): grupo deoxicorticosterona (grupo DOCA); grupo deoxicorticosterona e Imatinib (grupo DOCA IMA). A Pressão Arterial Sistólica (PAS) foi medida quinzenalmente. Foi estudada a porção apical do ventrículo esquerdo. Foram empregados: coloração vermelho sirius, coloração de hematoxilina-eosina, imuno-histoquímica e ensaio de western blot. RESULTADOS: A PAS nos grupos DOCA e IMA+DOCA foi maior que no grupo CON nos dias 14 e 28. Os animais do grupo DOCA apresentaram fibrose intersticial e perivascular grave no dia 28, e as expressões de PI, PIII, tenascina-C e fibronectina foram significativamente maiores que nos grupos DOCA+IMA e CON. Quando comparados com o grupo CON, os grupos DOCA e DOCA+IMA apresentaram resposta inflamatória de tecido miocárdico e infiltração de monócitos/macrófagos de diferentes graus. As expressões proteicas do PDGF-A, PDGF-C e PDGFRα foram significativamente maiores nos grupos DOCA e DOCA+IMA que no grupo CON, mas a expressão proteica do p-PDGFRα no grupo DOCA+IMA foi menor que no DOCA. CONCLUSÃO: O Imatinib pode exercer efeitos inibitórios sobre a fibrose miocárdica em ratos com hipertensão induzida por DOCA/sal, os quais podem ser atribuídos à inibição da atividade do PDGFR-α.

BACKGROUND: Imatinib is a tyrosine kinase receptor inhibitor that has been confirmed to exert inhibitory effect on the platelet derived growth factor PDGF receptor (PDGFRα and PDGFRβ) activity. OBJECTIVE: To investigate the protective effect of imatinib on the myocardial fibrosis in deoxycorticosterone-acetate (DOCA)/salt induced hypertensive rats. METHODS: Sixty male uninephrectomized Sprague-Dawley rats were assigned to three groups: control rats (CON group); deoxycorticosterone group (DOCA group); deoxycorticosterone and imatinib group (DOCA+IMA group). Systolic blood pressure (SBP) was measured biweekly. The apical portion of the left ventricle was studied. Sirius-Red staining, Hematoxylin-Eosin staining, immunohistochemistry and Western blot assay were employed. RESULTS: SBP in the DOCA group and DOCA+IMA group was higher than that in the CON group on day 14 and 28. Animals in the DOCA group showed severe interstitial and perivascular fibrosis on day 28, and the expressions of PI, PIII, tenascin-C and fibronectin were significantly higher than those in the DOCA+IMA group and CON group. When compared with the CON group, myocardial tissue inflammatory response and monocyte/macrophage infiltration of different degrees were observed in the DOCA group and DOCA+IMA group. Protein expressions of PDGF-A, PDGF-C and PDGFRα were signiflcantly higher in the DOCA and DOCA+IMA groups than those in the CON group, but the p-PDGFRα protein expression in the DOCA+IMA group was lower than that in the DOCA group. CONCLUSION: Imatinib can exert inhibitory effects on myocardial fibrosis in DOCA/salt induced hypertensive rats, which may be attributed to the inhibition of PDGFR-α activity.

Infecção genital e marcadores preditivos do parto prematuro / Genital infection and preterm birth predictive markers

Apesar dos avanços na área da Obstetrícia, a prematuridade ainda é a principal causa de morbidade e mortalidade neonatal. Esforços continuam sendo feitos na busca de marcadores que possam predizer tal evento e, assim, evitar o parto prematuro. A presente revisão abordou a relação da infecção genital com a prematuridade e sua influência nos resultados dos principais indicadores preditivos do parto prematuro, bem como a eficácia da antibioticoterapia. Os estudos revisados sugerem que o tratamento da vaginose bacteriana relaciona-se a resultados satisfatórios para a prevenção da prematuridade somente se realizado na primeira metade da gestação. A alteração dos testes preditivos não indica, por si só, o uso de antibióticos.

Despite advances in the field of Obstetrics, prematurity is still responsible for the leading cause of neonatal morbidity and mortality. Efforts are still being made in the search for markers that can predict such an event and thus prevent premature delivery. This review explores the relationship of genital infection with preterm birth and its influence on the results of the main preterm birth predictive markers, as well as the effectiveness of antibiotics. The revised studies suggest that treatment of bacterial vaginosis relates to satisfactory results for the prevention of preterm performed only if the first half of pregnancy. The change of predictive tests not indicates, by itself, the use of antibiotics.

Fibronectina fetal como predictor del trabajo de parto en mujeres mexicanas / Fetal fibronectin as a predictor of labor in Mexican women

Introducción: la presencia de fibronectina fetal en secreciones vaginales ha sido considerada como un predictor de trabajo de parto en embarazo de término y de pretérmino. Objetivo: evaluar la validez predictiva de la fibronectina en embarazadas que acudieron al Hospital General SSH de Pachuca, México. Metodología: se incluyeron pacientes embarazadas que acudieron al hospital para control de su embarazo. Se determinó la fibronectina fetal en todas las participantes y se dio un seguimiento hasta el inicio del trabajo de parto. Resultados: participaron un total de 148 pacientes, siendo un grupo con 53 pacientes con menos de 37 semanas de gestación (SG) y otro grupo con 95 pacientes con 37 ó más SG. En general, la prueba mostró una sensibilidad promedio de 72,5 por ciento y una especificidad promedio de 82,9 por ciento para ambos grupos. Conclusión: sobre la base de los resultados obtenidos, recomendamos utilizar la prueba de fibronectina en embarazadas a partir de las 32 semanas de gestación, tanto en los servicios de urgencias como de consulta externa.

Background: The presence of fetal fibronectin in vaginal secretions has been regarded as a predictor of labor in pregnant term and preterm. Objective: For this reason the purpose of this study was to evaluate the predictive validity of fibronectin in pregnant women who attended the General Hospital SSH Pachuca, Hidalgo, Mexico. Methodology: We included pregnant patients admitted to hospital for pregnancy control. Fetal fibronectin was determined in all participants and then followed until the onset of labor. Results: A total of 148 patients participated. One group with 53 patients less than 37 weeks gestation, and another group of 95 patients with 37 or more weeks gestation. In general, the test showed an average sensitivity of 72.5 percent and specificity 82.9 percent average for both groups. Conclusion: Based on these results, we recommend using fibronectin test in pregnant women after 32 weeks of gestation, both in emergency departments and outpatient clinics.

Effect of ProRoot MTA, Portland cement, and amalgam on the expression of fibronectin, collagen I, and TGFβ by human periodontal ligament fibroblasts in vitro.

Context: Today many materials have been introduced for root-end filling materials. One of them is mineral trioxide aggregate (MTA) that is mentioned as a gold standard. Aims: The purpose of this in vitro study was to evaluate the reaction of human periodontal ligament fibroblasts to the root-end filling materials, such as ProRoot MTA, Portland cement, and amalgam. Settings and Design: Eight impacted teeth were extracted in aseptic condition. The tissues around the roots were used to obtain fibroblast cells. After cell proliferation, they were cultured in the chamber slides and the extracts of the materials were added to the wells. Materials and Methods: Immunocytochemical method for measuring the expression of Fibronectin, collagen I and transforming growth factor beta (TGF®) was performed by Olysia Bioreport Imaging Software. Statistical Analysis Used: The results were analyzed by SPSS 13.0 and Tukey post hoc test with P<0.05 as the limit of significance. Results: Collagen expression in MTA specimens was higher than the other groups in 24 h significantly. After 48 h, the Portland cement group showed the most expression of collagen significantly and after 1 week, Portland cement and MTA groups had the most expression of collagen but there was no significant difference between these 2 groups. After 1 week, the Portland cement group demonstrated a higher amount of TGF® and fibronectin. Conclusions: The results suggest that Portland cement can be used as a less expensive root filling material with low toxicity. It has better effects than amalgam on the fibroblasts.

Estudo da virulência, adesão e características fenotípicas de isolados do complexo Sporothrix / Study of virulence, adhesion and phenotypic characteristics of isolates of Sporothrix complex

A esporotricose é uma doença micótica, infecciosa e crônica, que envolve o tecido cutâneo e subcutâneo, e que pode afetar seres humanos e animais. Esta micose sempre foi atribuída a um único patógeno, o Sporothrix schenckii, um fundo termodimórfico, que cresce como levedura a 37 oC e como micélio à temperatura ambiente. No entanto, nos últimos anos, foi demonstrado que isolados identificados como S. schenckii apresentavam grande variabilidade genética, sugerindo que esta táxon consiste em um complexo de espécies. Esta doença é causada pela implantação traumática do patógeno fúngico, porém os mecanismos de invasão e disseminação deste microorganismo, bem como as moléculas envolvidas nestes processos, ainda são pouco conhecidos. Com base nessas informações, este trabalho visa identificar moléculas de superfície deste patógeno envolvidas na interação deste fungo com proteínas matriciais, bem como analisar diferenças fenotípicas entre espécies do denominado complexo Sporothrix. Foram utilizados, neste estudo, cinco isolados de Sporothrix spp., sendo três isolados clínicos, um isolado ambiental e um isolado de gato. A virulência de cada isolado foi comparada à capacidade adesiva à proteína matricial fibronectina. Foi observado que os isolados com amior capacidade infectiva eram os que apresentavam maior capacidade adesiva à fibronectina. Verificamos então a expressão de adesinas para fibronectina na superfície de cada isolado, por Western blot, e observamos que os isolados mais virulentos e com maior capacidade adesiva expressavam mais adesinas para fibronectina. Bandas reativas com o anticorpo monoclonal contra adesina gp70 (mAb P6E7) foram reveladas nos extratos de parede celular dos isolados estudados. Análises por microscopia confocal revelaram a colocalização da gp70 com a adesina para fibronectina na superfície dos isolados. Análises filogenéticas demonstraram que os isolados estudados possuíam diferenças genotípicas capazes de agrupá-los em duas espécies...

Sporotrichosis is a chronic and infectious diseases that involves the cutaneous and subcutaneous tissue, which can affect humans and animals. This mycosis has always been attributed to a single pathogen, the Sporothrix schenckii, a dimorphic fungus, that grows as yeast at 37 oC and as mycelia at room temperature. However, in recent years, some isolates identifies as S. schenckii showed considerable genetic variability, sugesting that this taxon consists of a complex of species. This disease is caused by the traumatic inoculation of the fungal pathogen, however, the molecules involved in the invasion and dissemination of this microorganism are still poorly understood. The aim of this study is to identify surface molecules involved in the interaction of this fungus with extracellular matrix proteins and to examine phenotypic differences between species in the Sporothrix complex. Five isolates were used throughout this study, three clinical isolates, an environmental and one cat isolate. The virulence of each isolate was compared to the adhesive capacity to fibronectin. We observed that the most virulent isolates exhibited the higher capacity to interact with fibronectin. The expression of adhesins for fibronectin on the surface of each isolate was verified by Western blot. This analysis showed that the most virulent isolates expressed more fibronectin adhesins than the avirulent ones. Positive bands for the monoclonal antibody raised against gp70 adhesin (mAb P6E7) were revealed in cell wall extracts of the isolates studied. Confocal microscopy confirmed the colocalization of fibronectin and mAb P6E7 on the yeast cell surface. Molecular analysis showed genotypic differences between isolates used in this study, that can cluster than them into two species, S. schenckii and S. brasiliensis. This phylogenetic analysis revealed that the avirulent isolate was S. brasiliensis and not S. schenckii as previously thought. This new data led us o determine whether...

Utility of fibronectin in immuocytochemial differentiation of reactive mesothelial cells from metastatic malignant cells in serous effusions.

Cytological evaluation of serous effusions often poses difficulties to the pathologists. This study was designed to assess the utility of fibronectin as a mesothelial cell marker and evaluate its use along with carcinoembryonic antigen (CEA) as a short panel to aid in the differentiation of reactive mesothelial cells from metastatic adenocarcinoma cells in serous effusions. Forty serous effusion cases from clinically benign cases and forty from clinically malignant cases with a known primary were included in the study. After routine cytological evaluation, the cases were categorized as benign, suspicious and malignant. All the cases were studied for fibronectin and CEA immunostaining using APAAP technique. In the present study, fibronectin emerged as a 100% specific and 93.4% sensitive marker for mesothelial cells. CEA immunostaining was seen in 80% cases of metastatic adenocarcinoma in serous effusions. Mesothelial cells did not show any CEA positivity. Short panel of CEA and fibronectin aided in reaching a correct diagnosis in three out of five cytologically suspicious cases.

Longitud cervical y fibronectina en el síntoma de parto prematuro / Cervical length and fetal fibronectin testing for the prediction of preterm labour

Antecedentes: En Chile el parto prematuro es la principal causa de morbilidad y mortalidad perinatal. Objetivo: Evaluar la utilidad clínica de la longitud cervical y la detección de fibronectina, en la predicción de parto prematuro. Método: Estudio observacional prospectivo, de corte transversal en 86 pacientes que consultaron por síntomas de parto prematuro (SPP), con membranas indemnes y edad gestacional entre las 22 y 34 semanas. El grupo estudio formado por gestantes con parto prematuro dentro de los 7 días. Al ingreso se tomó muestra cervical para fibronectina y se midió longitud cervical mediante ultrasonografía transvaginal. Resultados: De las 86 gestantes estudiadas, en 13 (15 por ciento) se produjo el parto dentro de los 7 días. La curva ROC mostró un área bajo la curva para longitud cervical de 0,475 y sensibilidad 31 por ciento (4/13), especificidad 96 por ciento (70/73), valor predictivo positivo 57 por ciento (4/7) y valor predictivo negativo 89 por ciento (70/79), riesgo relativo positivo 5,0 y riesgo relativo negativo 0,47 (p=0,014). El área bajo la curva para fibronectina fue 0,92 con una sensibilidad 77 por ciento (10/13), especificidad 90 por ciento (66/73), valor predictivo positivo (10/17) 59 por ciento y valor predictivo negativo 96 por ciento (66/69), riesgo relativo positivo 13,48 y riesgo relativo negativo 0,43 (p<0,001). Conclusión: En pacientes con síntoma de parto prematuro, cuello no remodelado y fibronectina cervical negativa, se asocian con bajo riesgo de parto prematuro. La fibronectina positiva, fue mejor predictor de parto prematuro dentro de los siete días y la longitud cervical fue superior para descartarlo.

Background: In Chile the preterm labor is the main cause of perinatal morbidity and mortality. Objective: To evaluate the clinical utility of the cervical length and fibronectin determination in predicting preterm delivery. Method: A prospective observational and transactional study was made, in 86 patients with preterm labor symptoms and gestational age between 22 and 34 weeks. The study group included patients with labor and delivery within a week of admission. From each patient a cervico-vaginal specimen was obtained to measure fibronectin and transvaginal ultrasonographic evaluation of the uterine cervix. Results: From 86 patients admitted to the study, thirteen had preterm labor (15 percent). The ROC curve showed an area under curve for cervical length of 0.475 and sensitivity 31 percent (4/13), specificity 96 percent (70/73), positive predictive value 57 percent(4/13), negative predictive value 88 percent (70/79), positive relative risk was 5.0, and the negative relative risk 0.47 (p=0.014); for fibronectin was 0.92 and sensitivity 77 percent (10/13), specificity 90 percent (66/73), positive predictive value 59 percent (10/17), negative predictive value 96 percent (66/69), positive relative riskof 13.48, negative relative risk 0.43 (p=0.014). Conclusion: In patients with premature labor symptoms, the cervical lengths none shortened and negative fibronectin result were associated with a low risk of having a preterm labor. Positive fibronectin was a better predictor of premature birth in a week of admission and the cervical length was the best in ruling out the occurrence of a premature birth.

Caracterização imuno-histoquímica de lesão híbrida de ameloblastoma desmoplásico e convencional / Hybrid lesion of desmoplastic and conventional ameloblastoma: immunohistochemical aspects

O ameloblastoma é uma neoplasia odontogênica benigna comumente encontrada nos ossos maxilares. Histologicamente, mostra diversos padrões, incluindo a ameloblastoma plexiforme e folicular. Quando estes padrões histológicos coexistem com um ameloblastoma que exibe abundante desmoplasia, são então denominados de lesão æhíbrida" de ameloblastoma desmoplásico e convencional. No presente trabalho, nos propomos a relatar um caso de lesão híbrida de ameloblastoma desmoplásico e convencional destacando os aspectos imuno-histoquímicos relativos a expressão das proteínas da matriz extracelular (tenascina, fibronectina e colágeno I).

Ameloblastoma is a benign epithelial odontogenic tumor and is the most commonly encountered odontogenic tumor in the jaws. Histologically, ameloblastomas occur in different patterns, including plexiform pattern and follicular pattern. "Hybrid " lesion of ameloblastoma is a tumor variant in which histologically, areas of follicular or plexiform ameloblastoma coexist with characteristic areas of ameloblastoma exhibiting pronounced stromal desmoplasia (desmoplastic ameloblastoma). The purpose of this article is to present a case of "hybrid" lesion of desmoplastic ameloblastoma (AD) and conventional, and investigate extracellular matrix proteins such as tenascin, fibronectin, and type I collagen.

Comparison between fetal fibronectin and cervical index in early detection of preterm labor

The present study will compare between two methods of diagnosis of preterm labor, which are, Transvaginal Ultrasonography [for cervical index] and fetal fibronectin in cervicovaginal secretions and to evaluate their rules in early detection of preterm labor. This is a prospective study carried out at Obstetrics and Gynecology Department, Suez Canal University Hospital, Ismailia Governor ate [2001-2002]. The study was carried out on Patients attending the department of Obstetric and Gynecology who were pregnant >/= 24 - < 37 weeks of gestation and at risk of preterm labor. The material of the study included 50 cases divided into two groups according to strict inclusion criteria. Each case was subjected to thorough questionnaire obstetric examination, determination of fibronectin level in cervico-vaginal secretions and estimation of the cervical index. The results of the study showed the superiority of the fibronectin estimation for the earlier detection of cases subjected to impending preterm delivery compared to cervical index. The conclusion of the study is that the combined use of cervical index and fibronectin assay give better anticipation to cases with impending preterm delivery

Alterations in proteins of bone marrow extracellular matrix in undernourished mice

The objective of the present study was to determine the effect of protein malnutrition on the glycoprotein content of bone marrow extracellular matrix (ECM). Two-month-old male Swiss mice were submitted to protein malnutrition with a low-protein diet containing 4 percent casein as compared to 20 percent casein in the control diet. When the experimental group had attained a 20 percent loss of their original body weight, we extracted the ECM proteins from bone marrow with PBS buffer, and analyzed ECM samples by SDS-PAGE (7.5 percent) and ECL Western blotting. Quantitative differences were observed between control and experimental groups. Bone marrow ECM from undernourished mice had greater amounts of extractable fibronectin (1.6-fold increase) and laminin (4.8-fold increase) when compared to the control group. These results suggest an association between fluctuations in the composition of the hematopoietic microenvironment and altered hematopoiesis observed in undernourished mice

Reparaçäo miocárdica quando em uso de baixa dosagem e longa duraçäo de inibidor da síntese do óxido nítrico: miofibroblastos, colágeno tipo III e fibronectina / Myocardial repair with long-term and low-dose administration of a nitric oxide synthesis inhibitor: myofibroblasts, type III collagen and fibronectin

Objetivo - Estudar o processo de reparação miocárdica em ratos hipertensos sob inibição da síntese do óxido nítrico. Métodos - Foram estudados dois grupos de animais (controle e L-NAME 12 mg/kg/dia). A presença de colágeno tipo III, fibronectina e células contendo a-cima de músculo liso foi examinada por técnica imuno-histoquímica. Resultados - A fibronectina foi observada em ambas as lesões, recentes e tardias, enquanto que o colágeno tipo III foi observado, principalmente, nas áreas de cicatrização incompleta entre os miócitos e em torno dos ramos das artérias coronárias. As áreas de lesões recentes e tardias mostraram a presença de células fusiformes. A análise imuno-histoquímica demonstrou positividade para a actina de músculo liso nestes células. Conclusão - As células que expressam alpha-actina de músculo liso no miocárdio de ratos hipertensos estão associadas a acúmulo de colágeno tipo III e de fibronectina nas áreas de lesão.

Detection of fetal fibronection in twin pregnancies in relation to gestational age

Context: The presence of fetal fibronectin in the cervix or vagina has been investigated as a possible marker for the risk of preterm birth. Fetal fibronectin in cervical fluid can provide direct evidence of pathologic changes at the interface of fetal and maternal tissues. Objective: To evaluate the presence of fetal fibronectin as a predictor of premature delivery in twin pregnancies in relation to gestational age. Design: Acuracy study. Setting: University referral unit. Participants: 52 pregnat women with twin pregnancies and gestacional age of between 24 and 34 weeks. Main measurements: Sensivity, specifity, predictive values and relative risk rations of the correlation between fetal fibronectin and preterm birth before 34 and 37 weeks using an immediate-reading membrane test on cervicovaginal secretions obtained from participants. Result: The sensitivity varied between 66.7 per cent and 85.7 per cent, whereas the specificity was from 58.3 per cent to 81.8 per cent according to gestacional age at the time of sampling. The relative risk of spontaneous preterm birth after a positive fetal fibronectin test, as compared with a negative fetal fibronectin test, rose from 2.8 at 24-26 weeks to 4.1 at 27-30 weeks. Analyses of the risk of delivery before 34 weeks were not statistically significant. Conclusion: Fetal fibronectin in the cervicovaginal secretions of patients with twin pregnancies is a useful tool for the early identification of twin pregnancies likely to delivery before 37 weeks. However, the clinical value of the fibronectin test is limited because of low indices for prediction of delivery before 34 weeks. The best period for performing the fibronectin test in twin pregnancies to predict preterm delivery is between 27 and 30 weeks.

Fetal Fibronectin in cervical and vaginal secretions as a predictor of preterm delivery

To evaluate the detection of fetal fibronection in cervico-vaginal secretions of patients with symptoms suggestive of preterm labour, as a predictor of preterm birth, 235 women presenting to the delivery suite between 24 to 37 weeks of gestation were selected. Speciemens of cervico-vaginal secretions were collected and fetal fibronectin levels were measured by >/= ELISA. Samples were considered positive if the level was >/= 005 ug/ml. It was found that fetal fibronection predicted preterm labour with positive predictive value of 80.76%, negative predictive value of 89.61%, sensitivity of 68.85% and specificity of 94.25%. Analysis of the data from women at less than 34 weeks of gestation showed similar results. Of the negative fetal fibronectin results obtained upon admission, 96.75% of women delivered after 7 days. It was concluded that fetal fibronectin appears as a promising marker for predicting preterm delivery in high risk women. However, only when we fully understand how the mechanisms underlying the release and their initiation are related to the processes leading to labour, will the full potential of the test be realized