The role of Osterix protein in the pathogenesis of peripheral ossifying fibroma

Abstract Peripheral ossifying fibroma (POF) is a reactive lesion of oral tissues, associated with local factors such as trauma or presence of dental biofilm. POF treatment consists of curettage of the lesion combined with root scaling of adjacent teeth and/or removal of other sources of irritants. This study aimed to analyze the clinical and pathological features of POF and to investigate the immunoexpression of Osterix and STRO-1 proteins. Data such as age, gender, and size were obtained from 30 cases of POF. Microscopic features were assessed by conventional light microscopy using hematoxylin-eosin staining and immunohistochemical markers, and by polarized light microscopy using Picrosirius red staining. The age range was 11-70 years and 70% of the patients were female. Moreover, the size of POF varied from 0.2 to 5.0 cm; in 43.33% of the cases, the mineralized content consisted exclusively of bony trabeculae. The immunohistochemical analysis showed nuclear staining for Osterix in 63% and for STRO-1 in 20% of the cases. Mature collagen fibers were observed in mineralized tissue in 76.67% of the cases. The clinical and microscopic features observed were in agreement with those described in the literature. Osterix was overexpressed, while STRO-1 was poorly expressed. Osterix was expressed particularly in cells entrapped in and around mineralized tissue, indicating the presence of a stimulus that triggers the differentiation of these cells into osteoblasts or cementoblasts, i.e., cells that produce mineralized tissue. Based on our results, Osterix may play a role in the pathogenesis of POF.

Fetal hematopoietic stem cells express MFG-E8 during mouse embryogenesis

The milk fat globule-EGF-factor 8 protein (MFG-E8) has been identified in various tissues, where it has an important role in intercellular interactions, cellular migration, and neovascularization. Previous studies showed that MFG-E8 is expressed in different cell types under normal and pathophysiological conditions, but its expression in hematopoietic stem cells (HSCs) during hematopoiesis has not been reported. In the present study, we investigated MFG-E8 expression in multiple hematopoietic tissues at different stages of mouse embryogenesis. Using immunohistochemistry, we showed that MFG-E8 was specifically expressed in CD34+ HSCs at all hematopoietic sites, including the yolk sac, aorta-gonad-mesonephros region, placenta and fetal liver, during embryogenesis. Fluorescence-activated cell sorting and polymerase chain reaction analyses demonstrated that CD34+ cells, purified from the fetal liver, expressed additional HSC markers, c-Kit and Sca-1, and that these CD34+ cells, but not CD34- cells, highly expressed MFG-E8. We also found that MFG-E8 was not expressed in HSCs in adult mouse bone marrow, and that its expression was confined to F4/80+ macrophages. Together, this study demonstrates, for the first time, that MFG-8 is expressed in fetal HSC populations, and that MFG-E8 may have a role in embryonic hematopoiesis.

Bone marrow mesenchymal stem cells overexpressing human basic fibroblast growth factor increase vasculogenesis in ischemic rats

Administration or expression of growth factors, as well as implantation of autologous bone marrow cells, promote in vivo angiogenesis. This study investigated the angiogenic potential of combining both approaches through the allogenic transplantation of bone marrow-derived mesenchymal stem cells (MSCs) expressing human basic fibroblast growth factor (hbFGF). After establishing a hind limb ischemia model in Sprague Dawley rats, the animals were randomly divided into four treatment groups: MSCs expressing green fluorescent protein (GFP-MSC), MSCs expressing hbFGF (hbFGF-MSC), MSC controls, and phosphate-buffered saline (PBS) controls. After 2 weeks, MSC survival and differentiation, hbFGF and vascular endothelial growth factor (VEGF) expression, and microvessel density of ischemic muscles were determined. Stable hbFGF expression was observed in the hbFGF-MSC group after 2 weeks. More hbFGF-MSCs than GFP-MSCs survived and differentiated into vascular endothelial cells (P<0.001); however, their differentiation rates were similar. Moreover, allogenic transplantation of hbFGF-MSCs increased VEGF expression (P=0.008) and microvessel density (P<0.001). Transplantation of hbFGF-expressing MSCs promoted angiogenesis in an in vivo hind limb ischemia model by increasing the survival of transplanted cells that subsequently differentiated into vascular endothelial cells. This study showed the therapeutic potential of combining cell-based therapy with gene therapy to treat ischemic disease.

PGC and PSMA in prostate cancer diagnosis: tissue analysis from biopsy samples

Purpose The discovery of new diagnostic tools for the diagnosis of prostate cancer (PCa) has become an important field of research. In this study, we analyzed the diagnostic value of the expression of the pepsinogen C (PGC) and prostate-specific membrane antigen (PSMA) genes in tissue samples obtained from prostate biopsies. Materials and Methods This study was comprised of 51 consecutive patients who underwent transrectal ultrasound (TRUS)-guided prostate biopsies between January 2010 and March 2010. The biopsies were performed with 12 cores, and an additional core was randomly retrieved from the peripheral zone from each patient for study purposes. The expression of the PGC and PSMA genes was analyzed from the cDNA from the samples via the qRT-PCR technology. The expression patterns of patients with PCa were compared with those of patients without a PCa diagnosis. Results PSMA was overexpressed in only 43.4% of PCa cases, and PGC was overexpressed in 72.7% of cases. The median expression of PSMA was 1.5 times (0.1 to 43.9) and the median PGC expression was 8.7 times (0.1 to 50.0) the expression observed in prostatic tissue from TRUS-guided biopsies of normal patients. Analysis of patients with high-risk PCa indicated that PGC was overexpressed in 71.4% of cases (with a median expression of 10.6 times), and PSMA was overexpressed in only 35.7% of cases (with a median expression of 4.5 times). Among patients with low-risk PCa, PGC was also overexpressed in 71.4% of cases (with a median expression of 5.9 times), and PSMA was overexpressed in only 42.8% of cases (with a median expression of 2.5 times). Conclusions PGC gene expression is significantly higher in prostatic tissue in men affected by PCa when compared to normal prostates. Further analyses are necessary to confirm our results. .

Prognostic value of carcinoembryonic antigen distribution in tumor tissue of colorectal carcinoma / Valor prognóstico da distribuição do antígeno carcinoembriônico (CEA) no tecido neoplásico do carcinoma colorretal

CONTEXT: Carcinoembryonic antigen (CEA) can be detected in colorectal tumor tissue but its role in the survival of patients remains controversial. OBJECTIVE: To characterize the expression of tissue CEA using immunohistochemical staining in colorectal tumors and to analyze the relationship between this finding and preoperative plasmatic level of CEA, morphologic features and survival of patients operated with curative intent for colorectal carcinoma. METHOD: Forty-seven patients were included in the study: 18 (38.3 percent) males and 29 (61.7 percent) females, with a mean age of 67.8 ± 9.7 years (37 to 84 years). Immediately before laparotomy, pre-operative serum levels of CEA were obtained where normal levels were considered <2.5 ng/mL for non-smokers, and <5.0 ng/mL for smokers. CEA immunohistochemical studies were carried out using anti-human CEA monoclonal mouse antibody. The expression of immunostaining for each neoplasia was classified according to the pattern of CEA tissular distribution into apical or cytoplasmic. The variables considered for the statistical analysis were plasmatic preoperative CEA level, location of the lesion within the large intestine, lesion diameter, lymph node involvement, Duke's classification, vein invasion, grade of cellular differentiation, survival and pattern of CEA tissular distribution. The statistical models utilized were Spearman's correlation and the Mann-Whitney, Kruskal-Wallis and Student t tests. Patients' survival was analyzed using the Kaplan-Meier method. RESULTS: The mean preoperative CEA value was 15.4 ± 5.5 ng/mL (0.2 to 92.1 ng/mL). The neoplasm was located in the colon in 29 (61.7 percent) and in the rectum in 18 (38.3 percent) patients. Eight (17.0 percent) patients were classified as Duke's stage A, 22 (46.8 percent) as stage B and 17 (36.2 percent) as stage C. On immunohistochemical studies, the pattern of CEA tissular distribution was apical in 33 (70.2 percent) patients and cytoplasmic...

CONTEXTO: O antígeno carcinoembrionário (CEA) pode ser detectado no tecido do carcinoma colorretal, mas seu papel na sobrevivência dos doentes permanece controverso. OBJETIVO: Caracterizar a expressão do CEA tecidual com coloração imunoistoquímica na neoplasia colorretal e analisar a relação entre esse achado e os níveis plasmáticos pré-operatórios do CEA, aspectos morfológicos e a sobrevivência dos doentes operados com intenção curativa de carcinoma colorretal. MÉTODO: Quarenta e sete doentes foram incluídos neste estudo: 18 (38,3 por cento) homens e 29 (61,7 por cento) mulheres, com média de idade de 67,8 ± 9,7 anos (37 to 84 anos). Imediatamente antes da laparotomia, foram obtidos os níveis plasmáticos pré-operatórios do CEA. Níveis séricos pré-operatórios normais de CEA foram considerados < 2,5 ng/mL para não-fumantes e <5,0 ng/mL para fumantes. O estudo imunoistoquímico do CEA foi realizado utilizando anticorpo monoclonal de rato anti-CEA humano. A expressão da imunocoloração de cada neoplasia foi classificada de acordo com o padrão de distribuição tecidual do CEA em apical ou citoplasmática. As variáveis consideradas para a análise estatística foram os níveis plasmáticos pré-operatórios do CEA, localização da lesão no intestino grosso, diâmetro da lesão, comprometimento dos linfonodos, classificação de Dukes, invasão venosa, grau de diferenciação celular, sobrevivência e padrão da distribuição tecidual do CEA. Os modelos estatísticos utilizados foram correlação de Spearman, teste de Mann-Whitney, teste de Kruskal-Wallis e teste t de Student. A sobrevivência dos doentes foi analisada utilizando-se o método de Kaplan-Meier. RESULTADOS: O valor médio de CEA pré-operatório foi de 15,4 ± 5,5 ng/mL (0,2 a 92,1 ng/mL). A neoplasia estava localizada no colo em 29 (61,7 por cento) e no reto em 18 (38,3 por cento) doentes. Oito (17,0 por cento) doentes foram classificados como estádio A de Dukes, 22 (46,8 por cento) como estádio B e 17...

Haematologic and immunophenotypic profile of acute myeloid leukemia: an experience of Tata Memorial Hospital.

OBJECTIVES: To study the hematologic and immunophenotypic profile of 260 cases of acute myeloid leukemia at diagnosis. MATERIAL AND METHODS: This is a retrospective analysis of 260 cases of AML diagnosed at our institution between 1998 and 2000. Diagnosis was based on peripheral blood and bone marrow examination for morphology cytochemistry and immunophenotypic studies. SPSS software package, version 10, was used for statistical analysis. RESULTS: Seventy-six percent of our cases were adults. The age of the patients ranged from one year to 78 years with a median age of 27.2 years. There were 187 males and 73 females. The commonest FAB subtype, in both children and adults, was AML-M2. The highest WBC counts were seen in AML-M1 and the lowest in AML-M3 (10-97 x 10(9)/L, mean 53.8 x 10(9)/L). The mean values and range for hemoglobin was 6.8 gm/l (1.8 gm/l to 9.2 gm/l), platelet count 63.3 x 10(9)/L (32-83 x 10(9)/L), peripheral blood blasts 41.4% (5 to 77%) and bone marrow blasts 57.6% (34-96%). Myeloperoxidase positivity was highest in the M1, M2 and M3 subtypes. CD13 and CD33 were the most useful markers in the diagnosis of AML. CD14 and CD36 were most often seen in monocytic (38%) and myelomonocytic (44%) leukemias. Lymphoid antigen expression was seen in 15% of cases. CD7 expression was the commonest (11%). CONCLUSION: AML accounted for 39.8% of all acute leukemias at this institution. The most common subtype was AML-M2. Myeloperoxidase stain was a useful tool in the diagnosis of myeloid leukemias. CD13 and CD33 were the most diagnostic myeloid markers.

Seroprevalencia de anti-VHC, coexistencia de AG-HBS, anti HBC y HIV, and algunos factores de riesgo en donantes anti VHC / The anti-HCV seroprevalence, the coexistence of AG-HBS, anti-HBC and VIH, and some risk factors in positive anti-HCV donors

La seroprevalencia de Anti-VHC, la coexistencia de Ag-HBs, Anti-HBc, HIV y algunos factores de riesgo en donantes Anti-VHC positivo del Banco de Sangre "Jesús Boada Boada" de Barquisimeto durante enero-junio de 1999, se determinó a través de un estudio descriptivo transversal,donde la población y muestra estuvo conformada por el total de donantes voluntarios de sangre (3381), de los cuales 47 resultaron Anti-VHC positivo según la prueba ABBOTT HCV EIA 3.0. Se encontró una seroprevalencia de 1,21 por ciento; el 17.02 por ciento resultaron positivos también para Anti-HBc y 4.26 por ciento tienen positividad simultanea Ag-Hbs y Anti-HBc. Se realizó a 27 donantes voluntarios seropositivos visitas domiciliarias para citarlos al Ambulatorio Urbano tipo II "Pueblo Nuevo" donde se aplicó una encuesta estructural mediante la entrevista. el 88,89 por ciento pertenecen al sexo masculino, el 77,78 por ciento se ubica entre los 15 y 35 años; entre los principales factores de riesgo se encontró que el 81,48 por ciento estuvo en contacto con agujas e instrumental no estéril ó de esterilidad dudosa, 59.09 por ciento tuvo heridas accidentales en barberías, el 76 por ciento con vida sexual activa 3 ó más parejas sexuales y el 100 por ciento son heterosexuales, con ocupación de bajo riesgo para adquirir la infección, desconocen la seropositividad de sus parejas sexuales y del grupo familiar con el cual conviven

Differential expression of syntaxin-1 and synaptophysin in the developing and adult human retina.

Synaptophysin and syntaxin-1 are membrane proteins that associate with synaptic vesicles and presynaptic active zones at nerve endings, respectively. The former is known to be a good marker of synaptogenesis; this aspect, however, is not clear with syntaxin-1. In this study, the expression of both proteins was examined in the developing human retina and compared with their distribution in postnatal to adult retinas, by immunohistochemistry. In the inner plexiform layer, both were expressed simultaneously at 11-12 weeks of gestation, when synaptogenesis reportedly begins in the central retina. In the outer plexiform layer, however, the immunoreactivities were prominent by 16 weeks of gestation. Their expression in both plexiform layers followed a centre-to-periphery gradient. The immunoreactivities for both proteins were found in the immature photoreceptor, amacrine and ganglion cells; however, synaptophysin was differentially localized in bipolar cells and their axons, and syntaxin was present in some horizontal cells. In postnatal-to-adult retinas, synaptophysin immunoreactivity was prominent in photo-receptor terminals lying in the outer plexiform layer; on the contrary, syntaxin-1 was present in a thin immunoreactive band in this layer. In the inner plexiform layer, however, both were homogeneously distributed. Our study suggests that (i) syntaxin-1 appears in parallel with synapse formation; (ii) synaptogenesis in the human retina might follow a centre-to-periphery gradient; (iii) syntaxin-1 is likely to be absent from ribbon synapses of the outer plexiform layer, but may occur at presynaptic terminals of photoreceptor and horizontal cells, as is apparent from its localization in these cells, which is hitherto unreported for any vertebrate retina.

Monoclonal antibodies which inhibit in vitro cytotoxicity of axenic Entamoeba histolytica.

A panel of 12 independent hybridoma cell lines secreting monoclonal antibodies to axenic E. histolytica (HM1) have been developed. A hybridoma cell line P4 C4 P2 F8 C8 (clone C8) produced monoclonal antibodies (MoAb C8) of IgG1 isotype which recognised a 29 KD surface associated antigen of amoebic trophozoites in Western immunoblot. Immunofluorescent probing with MoAb C8 employing live and acetone fixed amoebic trophozoites indicated 29 KD molecule on the surface plasma membrane of E. histolytica trophozoites. The MoAb C8 also agglutinated the live amoebic trophozoites. Pretreatment of amoebic trophozoites with anti 29 KD monoclonal antibody significantly (P less than 0.01) inhibited in vitro cytotoxicity of amoebic trophozoites to the cultured baby hamster kidney (BHK-21) cells. MoAb recognised a 29 KD molecule of E. histolytica trophozoites which mediated cytotoxic potentials of the parasite. The absence or variable degree of expression of cytotoxic 29 KD molecule may possibly serve as a marker to differentiate virulent/avirulent populations or strains of E. histolytica.

Identification and characterization of sex-linked proteins of Schistosoma mansoni

The proteins of adults worms (male and female) of two isolates (BH and RJ) of Shistosoma mansoni were extracted using Triton X-114 phase separation. The SDS-polyacrilamide gel electrophoresis profiles of the three phases (detergent, aqueous and insoluble proteins) obtained were compared after Coomassie blue and silver staining, surface radioiodination and Western blotting. No major differences were detected between the 2 isolates. Of the 25 or more proteins which partitioned into the detergent phase, only about 8 proteins could be surface radiodinated on live adult worms. A comparison was also made between the profiles of mael and females worms, isolated from bisexually infected mice. Two major female-specific and one male-specific band were detected by silver and/or Coomassie staining. The female bands, 32 KDa and 18 Kda, partitioned into the detergent and aqueous phase, respectively. The male-specific band of 42 KDa remained in the insoluble phase. Antigenic differences between male and females protins were detected by Western vlotting using a sera from infected Nectomys squamipes

Simultaneous identification of Trypanosoma cruzi surface and internal antigens reactive to different immunoglobulin classes (radio-immunoblotting)

Classes e subclasses de anticorpos apresentam diferentes funçöes, influenciando a resposta imune humoral de um hospedeiro, frente a um agente infeccioso. Na maioria dos sistemas, o alvo principal é representado pelos antígenos de membrana do parasita. Entretanto, a identificaçäo de antígenos de superficie de parasitas, reativos para classe (e subclasse) de imunoglobulinas que näo se ligam a proteína-A implica em imunoprecipitaçöes sucessivas, que levam a perda de antígenos e/ou reaçöes inespecíficas. Visando esse estudo, foi desenvolvida ua técnica denominada "radio-immunoblotting", através da qual a reatividade de imunoglobulinas de diferentes classes para antígenos de membrana (e/ou internos) foi analisada simultaneamente. O método constitui na marcaçäo prévia da superfície dos parasitas por radioiodaçäo, fracionamento dos polipeptídeos por SDS/PAGE, transferência das fraçöes para nitrocelulose, reaçäo com soros e conjugados anti-Igs - peroxidase a autroradiografia dos mesmos, a análise é feita comparando-se os antígenos comuns evidenciados na autoradiografia e nas tiras de nitrocelulose coradas com o substrato da peroxidase. Essa técnica foi utilizada para analisar antígenos de superfície de formas tripomastigotas de T. cruzi reativas para IgG, IgM e IgA provenientes de soros de pacientes com doença de Chagas na fase aguda. Obtiveram-se distintos padröes de reatividade para as diferentes classes de anticorpos provenientes de um mesmo soro humano

Assessment of putative tests for protective immunity to falciparum malaria.

Sera from clinically immune individuals comprising 10 hospitalised patients (Group I), 30 persons residing in a malaria endemic area in Thailand (Group II) and 8 persons from a hyperendemic area in Ivory Coast (Group III) were tested by the parasite growth inhibition (PGI), indirect fluorescent antibody test of ring-infected erythrocyte surface antigen (RESA-IFA), urease-ELISA and Western blot. Paired sera from patients recovering from malaria (Group IV) as well as sera from blood donors were also tested. In the PGI test, sera were tested against three uncloned isolates of P. falciparum comprising SO, I4 and AE9 (PGI-SO, PGI-I4 and PGI-AE9 respectively). When growth inhibition of greater than or equal to 30% against any one of the three isolates was considered positive, the positive rate for the combined Groups I, II and III was 78.7%. Further analysis showed that the positive rates for PGI-SO, PGI-I4 and PGI-AE9 were 63.8%, 59.5% and 59.5% respectively and were not significantly different (p greater than 0.05). Comparison between PGI-SO, PGI-I4 and PGI-AE9 activities of Groups I, II and III sera showed no significant differences in any comparison groups except with PGI-AE9 in which Group III sera were more frequently positive than Group II sera (p = 0.004). Follow-up of PGI-SO and PGI-AE9 activities in Group IV patients showed mostly a decrease or no change in the activities of the convalescent sera taken 63 days later. RESA-IFA positive rate in the combined Groups I, II and III sera was 91.7%. There were no significant differences either in the seropositive rates or in the geometric mean antibody titers (GMT) between Groups I, II and III sera. Follow-up in Group IV patients showed no change in antibody titers in 64% of cases, decrease and increase in titers in 29% and 7% of cases respectively. The urease-ELISA seropositive rate in the combined Groups (I, II and III) was 89.5% which is not significantly different from that of RESA-IFA (p greater than 0.05). Comparison between individual Groups (I, II and III) likewise showed no significant differences in both GMT and seropositive rates. Follow-up in Group IV sera showed either no change or a decrease in antibody titers in 55.6% and 44.4% of cases respectively.(ABSTRACT TRUNCATED AT 400 WORDS)

Receptor de membrana de linfócito T humano para eritrócitos de carneiro (CD2) / Membrane receptor of human T lymphocyte for sheep erythrocyte CD2

Esta revisäo trata inicialmente das características do receptor CD2 e suas funçöes em linfócitos T e da açäo dos anticorpos monoclonais dirigidos contra essa estrutura em linfócitos T. Finalmente säo feitas algumas consideraçöes sobre os receptores na forma solúvel

Marcadores tumorais / Tumor markers

Alteraçöes sangüineas e de componentes celulares indicam a presença de tumor. Estas alteraçöes, chamadas de "MARCACORES TUMORAIS", podem ter valor diagnóstico, prognóstico, evolutivo e terapêutico, em conjunto com a avaliaçäo clínica

Trypanosoma cruzi: circulating polysaccharide factors excreted in vitro and in vivo

Se detectó un factor antigénico, temoestable y soluble en ácido tricloroacético (TCA) en el sobrenadante de cultivo de epimatigotes de Trypanosoma cruzi en fase logarítmica y en los sueros de pacientes con enfermedad de Chagas aguda. Este antígeno fue revelado en la mambrana de superficie de fibroblastos de ratas cuando fueron infectados con trypomastigotes, utilizando la técnica de inmunofluorescencia indirecta. El sobrenadante de cultivo de epimastigotes en medio GLSH libre de células a 28-C fue acidificado a pH5, calentado a ebullición 30 min y luego precipitado con TCA, centrifugado y el sobrenadante tratado con éter de petróleo. La solución extraída fue dializada, concentrada, liofilizada y purificada por cromatografía de afinidad. El eluido fue liofilizado y este material fue denominado factor antígeno excretado (EF). Un antisuero hiperinmune contra estos factores antígenos fue obtenido en conejos. El 76,6% de los sueros correspondientes a pacientes con Chagas agudo demostraron poseer el EF por técnica de inmunodifusión doble (Tabla 1). Monocapas de fibroblastos de ratas infectados previamente con 3 ml de trypomastigotes en una concentración de 5 x 10**6 células/ml sometidas a una inmunofluorescencia indirecta con el antisuero hiperinmune demostraron la presencia del EF en las membranas celulares de fibroblastos infectados y no infectados..

Tumor associated proteins of rat skin tumor induced by 7,12-dimethylbenzaanthracene

The incidence of tumor and the time of expression, cellular localization and the molecular weight of tumor associated proteins of rat skin tumor induced by 7,12-dimethylbenz[a]anthracene (DMBA) with or without 12-O-tetradecanoyl-phorbol-13-acetate (TPA) were studied. The time of the development of skin tumors in 0.1% DMBA-TPA treated rats was significantly shorter than that in rats which were treated with DMBA alone. In the complete carcinogenesis case, papillomas developed more slowly and were less common and also squamous cell carcinomas appeared much later. From the analysis of the proteins of each experimental group by SDS-PAGE and two dimensional gel electrophoresis, at least three tumor associated proteins were identified (54kd, pl = 5.66; 27kd, pl = 5.85; 11kd, pl = 4.90). Also these proteins were found in rat dorsal skin from 14 days gestation to 21 days postpartum, and disappeared after 28 days. In conclusions, two stage skin carcinogenesis could be successfully demonstrated in Sprague-Dawley rats and abnormal proteins were produced in DMBA or DMBA-TPA induced skin tumor. The tumor associated proteins of skin tumor induced by DMBA or DMBA-TPA were appeared at the late initiation stage or early promotion stage, and they were localized in plasma membrane and were glycoproteins that are thought to be related to the epidermal differentiation process.