Effects of Syzygium aromaticum, Cinnamomum zeylanicum, and Salvia triloba extracts on proliferation and differentiation of dental pulp stem cells

Abstract Hypersensitivity, local irritative and cytotoxic effects are known for the chemical components of Syzygium aromaticum and Cinnamomum zeylanicum contained in dental materials. However, there is no intimate data in dentistry using the whole extracts of these plants and introducing new ones. Salvia triloba is a well-known anti-inflammatory plant that correspondingly could be used in several dental traumas. Objectives: We aimed to show and compare the effect of S. aromaticum, C. zeylanicum, and S. triloba extracts on dental pulp stem cells (DPSCs) proliferation, differentiation, and immune responses. Material and Methods: Using xCELLigence, a real time monitoring system, we obtained a growth curve of DPSCs with different concentrations of the Extracts. A dose of 10 μg/mL was the most efficient concentration for vitality. Osteogenic differentiation and anti-inflammatory activities were determined by using an ELISA Kit to detect early and late markers of differentiation. Results: The level of osteonectin (ON, early osteogenic marker) decreased, which indicated that the osteogenic differentiation may be accelerated with addition of extracts. However, the level of osteocalcin (OCN, late osteogenic marker and sign of calcium granulation) differed among the extracts, in which S. aromaticum presented the highest value, followed by S. triloba and C. zeylanicum. Surprisingly, the determined calcium granules were reduced in S. aromaticum and S. triloba. In response to tumor necrosis factor alpha (TNF-α), S. triloba-treated DPSCs showed the most reduced level of IL-6 cytokine level. We suggest C. zeylanicum as a promising osteogenic inducer and S. triloba as a potent anti-inflammatory agent, which could be used safely in biocomposite or scaffold fabrications for dentistry. Conclusions: Because calcium granule formation and cell viability play a critical role in hard tissue formation, S. aromaticum in dentistry should be strictly controlled, and the mechanism leading to reduced calcium granule formation should be identified.

CDX-2, MUC-2 and B-catenin as intestinal markers in pure mucinous carcinoma of the breast

BACKGROUND: Pure mucinous adenocarcinoma of the breast is a rare entity characterized by the production of variable amounts of mucin comprising 1% to 6% of breast carcinomas. Some mucinous adenocarcinomas have shown expression of intestinal differentiation markers such as MUC-2. This study examines the expression of intestinal differentiation markers in this type of breast carcinoma. RESULTS: Twenty-two cases of pure mucinous adenocarcinoma of the breast were assessed. Immunochemistry was performed for beta-catenin, CDX-2 and MUC-2. All cases were positive for B-catenin. MUC-2 positivity was observed in all cases; 63. 6% were 3 plus positive. All cases were negative for CDX-2. CONCLUSIONS: These results suggest that mucinous breast carcinomas express some markers of intestinal differentiation, such as MUC-2 and beta-catenin; however, future studies with a larger series of cases and using molecular techniques that help affirm these results are needed.

Induced maturation of hepatic progenitor cells in vitro

Hepatic progenitor cells (HPCs) are a potential cell source for liver cell transplantation but do not function like mature liver cells. We sought an effective and reliable method to induce HPC maturation. An immortalized HP14.5 albumin promoter-driven Gaussian luciferase (ALB-GLuc) cell line was established from HPCs isolated from fetal mouse liver of post coitus day 14.5 mice to investigate the effect of induction factors on ALB promoter. HP14.5 parental cells were cultured in DMEM with different combinations of 2% horse serum (HS), 0.1 µM dexamethasone (DEX), 10 ng/mL hepatic growth factor (HGF), and/or 20 ng/mL fibroblast growth factor 4 (FGF4). Trypan blue and crystal violet staining were used to assess cell proliferation with different induction conditions. Expression of hepatic markers was measured by semi-quantitative RT-PCR, Western blot, and immunofluorescence. Glycogen storage and metabolism were detected by periodic acid-Schiff and indocyanine green (ICG) staining. GLuc activity indicated ALB expression. The combination of 2% HS+0.1 µM Dex+10 ng/mL HGF+20 ng/mL FGF4 induced the highest ALB-GLuc activity. Cell proliferation decreased in 2% HS but increased by adding FGF4. Upon induction, and consistent with hepatocyte development, DLK, AFP, and CK19 expression decreased, while ALB, CK18, and UGT1A expression increased. The maturity markers tyrosine aminotransferase and apolipoprotein B were detected at days 3 and 6 post-induction, respectively. ICG uptake and glycogen synthesis were detectable at day 6 and increased over time. Therefore, we demonstrated that HPCs were induced to differentiate into functional mature hepatocytes in vitro, suggesting that factor-treated HPCs may be further explored as a means of liver cell transplantation.

Parasitic Helminth Cystatin Inhibits DSS-Induced Intestinal Inflammation Via IL-10+F4/80+ Macrophage Recruitment

Many immune down-regulatory molecules have been isolated from parasites, including cystatin (cystain protease inhibitor). In a previous study, we isolated and characterized Type I cystatin (CsStefin-1) of the liver fluke, Clonorchis sinensis. To investigate whether the CsStefin-1 might be a new host immune modulator, we induced intestinal inflammation in mice by dextran sodium sulfate (DSS) and treated them with recombinant CsStefin-1 (rCsStefin-1). The disease activity index (DAI) increased in DSS only-treated mice. In contrast, the DAI value was significantly reduced in rCsStefin-1-treated mice than DSS only-treated mice. In addition, the colon length of DSS only-treated mice was shorter than that of rCsStefin-1 treated mice. The secretion levels of IFN-gamma and TNF-alpha in the spleen and mesenteric lymph nodes (MLNs) were significantly increased by DSS treatment, but the level of TNF-alpha in MLNs was significantly decreased by rCsStefin-1 treatment. IL-10 production in both spleen and MLNs was significantly increased, and IL-10+F4/80+ macrophage cells were significantly increased in the spleen and MLNs of rCsStefin-1 treated mice after DSS treatment. In conclusion, rCsStefin-1 could reduce the intestinal inflammation occurring after DSS treatment, these effects might be related with recruitment of IL-10 secreting macrophages.

Leucemia de células dendríticas: a propósito de un caso / Dendritic leukemia cells: a purpose of a case

La leucemia de células dendríticas son patologías poco frecuentes, que involucran fundamentalmente a la piel, pero con una alta tendencia de metástasis. La inmunohistoquímica es una herramienta valiosa junto con la biopsia para el diagnóstico definitivo. Se presenta el caso de una paciente joven, quien acudió por presentar una lesión en la pierna izquierda de dimensiones y características únicas, cuyo diagnóstico requirió de la utilización de marcadores monoclonales específicos en la identificación de esta entidad.

The dendritic cell leukemias are pathologies very uncommon which involve mainly the skin, but with a high tendency of metastasis. The flow cytometric is a valuable tool together with tissue biopsy for a definitive diagnosis. We present the case of a young female who complained for a left leg lesion with unique dimensions and characteristics, in where the diagnostic process required the use of the specific monoclonal markers in the identification of this particular disease.

Estudo da expressão de galectina-3 em carcinomas de língua de camundongos / Study of the galectin-3 expression in tongue carcinomas of mice

INTRODUÇÃO: A galectina-3 (Gal-3) é uma lectina de mamíferos ligante de resíduos b-galactosídeos. Numerosos estudos têm mostrado que a Gal-3 apresenta importantes papéis na biologia tumoral, atuando em fenômenos como apoptose, metástase e transformação maligna. No entanto, em carcinomas de cabeça e pescoço, a real significância da sua expressão ainda é pouco compreendida. OBJETIVO: O objetivo deste estudo foi avaliar a expressão de Gal-3 em tumores de língua induzidos experimentalmente em camundongos desafiados com o carcinógeno 4-nitroquinolona-1-óxido (4NQO). MATERIAL E MÉTODOS: Quarenta e dois camundongos C57BL/6, machos, foram desafiados com 4NQO na água de beber por 16 semanas e sacrificados em diferentes períodos depois do tratamento. Após o sacrifício, as línguas foram removidas, processadas, coradas por hematoxilina e eosina (HE) e microscopicamente analisadas quanto à presença de carcinoma. Ensaio imuno-histoquímico para detecção do antígeno Gal-3 e análise descritiva da sua expressão nos tumores induzidos foram realizados. RESULTADOS: Ao final do experimento, foram produzidos 15 tumores. No tecido epitelial não-neoplásico, forte imunorreatividade foi observada apenas na camada parabasal. Nas camadas mais superficiais a intensidade de marcação foi mais fraca, e na camada basal variou de ausente a fraca. Todos os carcinomas bem diferenciados exibiram fraca marcação, exceto nas áreas queratinizantes. No único caso de carcinoma pouco diferenciado, forte imunorreatividade para Gal-3 foi observada. CONCLUSÃO: Nossos resultados descritivos mostram que a transformação maligna é acompanhada de redução da intensidade de expressão da Gal-3 e que o aumento da sua expressão com a perda da diferenciação neoplásica sugere a sua vinculação com agressividade tumoral.

BACKGROUND: Galectin-3 (Gal-3) is a b-galactoside-binding mammalian lectin. Numerous studies have demonstrated that Gal-3 plays an important role in tumor biology, acting in some events such as apoptosis, metastasis and malignant transformation. However, in carcinomas of head and neck, the real significance of Gal-3 expression requires a better understanding. OBJECTIVES: The aim of this paper was to evaluate Gal-3 expression in tongue carcinomas experimentally induced in mice challenged with carcinogen 4-nitroquinoline-1-oxide (4NQO). MATERIAL AND METHOD: Forty-two C57BL/6 male mice were challenged with 4NQO in drinking water for 16 weeks and killed at different periods after induction. In each period, their tongues were removed, routinely processed, stained with hematoxylin and eosin (H&E) and microscopically analyzed as to the presence of carcinoma. Immunohistochemical test for Gal-3 and a descriptive analysis of its expression in induced tumors were performed. RESULTS: By the end of the experiment, 15 tumors had been induced. In the non-neoplastic lingual epithelium, strong and weak immunoreactivity for Gal-3 was noted in parabasal and superficial layers, respectively. In the basal layer, Gal-3 expression varied from absent to weak. All the well-differentiated carcinomas showed weak immunoreactivity for Gal-3, except in keratinized areas. The only case of poorly differentiated squamous cell carcinoma indicated strong immunoreactivity for Gal-3. CONCLUSION: Our results show that malignant transformation is associated with reduced expression of Gal-3, whereas its increased expression in poorly differentiated carcinoma seems to be connected with tumor aggressiveness.

Expressäo da molécula CD5 na leucemia linfocítica crônica / Expression of molecule CD5 in chronic lymphocytic leukemia

O CD5 é um antígeno normalmente associado ao linfócito T que se expressa de forma aberrante na leucemia linfóide crônica de células B (LLC-B) e no linfoma de células do manto (LCM). Outras neoplasias crônicas de células B como a leucemia prolinfocítica (LPL-B) e "haity cell leukemia" (HCL), säo geralmente CD5 negativas ou expressam fracamente este antígeno. Neste trabalho foi investigada a expressäo do CD5 em 42 pacientes com doenças linfoproliferativas crônica de células B, com um painel de anticorpos monoclonais específicos para doenças linfoproliferativa crônica como: CD3, CD4, CD5, CD7, CD8, CD10, CD14, CD16/56, CD19, CD22, CD23, HLADR, CD45, anti cadeia pesada (IgH) e leves kappa e lambda das imunoglobulinas. Os parâmetros hematológicos foram obtidos por analisador hematológico automático e a citomorfologia pela microscopia óptica de filme sangüíneo corado pelo Leishmann. Informaçöes adicionais referentes a dados clínicos relacionados a estas doenças também foram coletados. Vinte e dois pacientes eram do sexo masculino e vinte do sexo feminino, tendo a idade variado de 48 a 80 anos de idade. A imunofenotipagem mostrou que 35 pacientes tinham LLC, 3 eram HCL, 3 tinham LPL-B e um caso diagnosticado como LCM. A linfadenopatia r a esplenomegalia foram os achados clínicos mais freqüentes, observados em 73,8 porcento e 71,4 porcento dos casos respectivamente. A expressäo do CD5 mostrou-se presente em todos os casos de LLC e LCM; nos casos de LPL-B este marcador apresentou-se fracamente positivo em um caso. Na HCL, estretanto, o CD5 foi negativo em todos os casos. Concluindo, estes dados demonstraram que a expressäo do CD5 é relevante na distinçäo entre LLC de outras neoplasias crônicas linfóides de célula B como a HCL e LPL-B, näo apresentando valor significativo no diagnóstico diferencial entre a LLC e LCM

Reproducibility of alkaline antigens of Leishmania major-like and Leishmania (V.) braziliensis evaluated by IgG-ELISA: comparison of antigens added of a protein inhibitor (PMSF) or not

Neste trabalho estudou-se o comportamento de 10 partidas de antigenos alcalinos de L. major-like e L. (V.) braziliensis adicionados ou nao de um inibidor de protease (PMSF) avaliados em tres dias consecutivos por meio de ELISA-IgG empregando soros padrao positivo de pacientes com diagnostico de leishmaniose mucocutanea previamente testados para a presenca de anticorpos anti-leishmania por ELISA. A analise estatistica mostrou que para o antigeno de L. (V.) braziliensis adicionado de PMSF nao houve diferenca significante entre as partidas ou dias de teste. Para o antigeno sem PMSF houve diferenca entre as partidas mas nao entre os dias de teste. Uma ANOVA bi-caudal mostrou diferencas entre os antigenos com e sem PMSF. Os antigenos de L. major-like preparados com e sem adicao de PMSF mostraram diferencas significativas entre as partidas; os tres dias de teste foram significativamente diferentes para o antigeno preparado com PMSF mas somente os dias 1 e 2 o foram com o antigeno sem adicao de inibidor...

Anticuerpos anticitoplasma de neutrófilo (ANCA): Cuál es su significado clínico? / Anticytoplasmis neutrophylic antibodies: What's its clinical relevance?

Los anticuerpos anticitoplasma de neutrófilo (ANCA) constituyen un grupo de inmunoglobinas dirigidas contra las enzimas contenidas en los gránulos de los polimorfonucleares neutrófilos y monocitos. Los ANCA son maracadores serológicos de diversas enfermedades vasculíticas de vasos pequeños y medianos, granulomatosas o no. Por inmunofluorescencia indirecta, se identifican 2 subtipos principales de ANCA: citoplasmático (C ANCA) y perinuclear (P ANCA). Los C ANCA son hallados principalmente en la vasculitis granulomatosa de wegener (E.W.). Se han detectado C ANCA en otros procesos vasculíticos y no vasculíticos: poliarteritis microscópica, enfermedad de Churg Strauss, panarteritis nodosa, síndrome de Sweet, fibroquistosis, fibrosis pulmonar, procesos infecciosos y neoplásicos y en pacientes VIH positivos. La participación de C ANCA en estos cuadros es variable e infrecuente. Los P ANCA se vinculan a vasculitis limitada al riñón y a vasculitis sistémicas no granulomatosas. Los P ANCA son marcadores menos sensibles y específicos. Se los encuentra asociados principalmente a poliarteritis microscópica (PM), Churg Strauss y vasculitis limitadas al riñón. Se presentan dos casos representativos ANCA positivos: una E.V. y una P.M. El verdadero valor de los ANCA como marcadores de enfermedades vasculíticas o como índices de su actividad no está definitivamente establecido, excepción hecha de la E.W. donde la especificidad para C ANCA es del 90 al 100 por ciento

A monoclonal antibody to common acute lymphoblastic leukemia antigen (CALLA) and its expression on several human tumor cell lines

We describe a newly-made murine monoclonal antibody to the common acute lymphoblastic leukemia antigen (CALLA), named SHB-10. The antigen detected by SHB-10 has a molecular weight of about 105 kDa. This antibody is very similar to that of conventional anti-CD10 Ab on indirect flowcytometric analysis using lymphoid malignant cell lines and peripheral lymphocytes of acute lymphoblastic leukemia (ALL) patients. The binding of anti-CD10 to Daudi cell and peripheral lymphocytes of ALL patients is blocked by SHB-10. Thus this monoclonal antibody is thought to detect the CALLA. The distribution of antigen detected by SHB-10 on several cell lines of neuroectodermal tumor and lymphoid malignancy was analysed and a slight difference in their cell surface expression is observed when compared with that by conventional anti-CD10. Further biochemical analysis is now under way for a better characterization of this antigen.

Prevalencia de los marcadores serológicos del virus de la hepatitis B y respuesta a la inmunización activa en un instituto de personas subnormales / Prevalence of serological hepatitis B markers and response to active immunization in an institution for mentally subnormal patients

En un instituto para personas con diversos niveles de deficiencia mental, y el personal encargado de su cuidado, se estudia la prevalencia de la infección con virus de hepatitis B (HBV). Un 70 por ciento de los internados mostró uno o más marcadores indicadores de infección con HBV, mientras que el personal mostró un nivel de prevalencia del 12 por ciento que compara con el de la población de donantes de sangre en la Argentina. Los resultados obtenidos en el personal de atención indican que éste tiene inclusive menor riesgo que los empleados hospitalarios tomados globalmente. Los indicadores epidemiológicos observados y el encuadre de la experiencia documentada muestran el alto riesgo de infección por Hepatitis en este sistema de interacción. En forma inmediata se instaló un programa de vacunación de los susceptibles (sin ningún marcador positivo) lográndose una respuesta de anticuerpos a los 330 días en el 100 por ciento de los internados. Como corolario se propone ajustar las normas nacionales de vacunación actuales, incluyendo en primera prioridad a los internos en asilos de este tipo, que actualmente figuran en segunda

Malignant histiocytosis: Clinicopathologic review of 18 cases with immunohistochemical study

We reviewed clinical and pathologic characteristics of 18 cases of malignant histiocytosis including 2 autopsy cases with special interest on their immunohistochemical characteristics. We report 3 cases of unusual immunohistochemical finding and postulate these cases may be a supportive evidence of the view that dendritic cells share common precursor with those of mononuclear phagocytic system. More accumulated cases in the future and careful analysis would be needed to answer the basic question about the origin of malignant histiocytosis.