Effects of transforming growth factor-ß1 on plasminogen activation in stem cells from the apical papilla: role of activating receptor-like kinase 5/Smad2 and mitogen-activated protein kinase kinase (MEK)/extracellular signal-regulated kinase (ERK) signalling.

AIM: To study the effects of TGF-ß1 on the plasminogen activation (PA) system of stem cells from the apical papilla (SCAP) and its signalling. METHODOLOGY: SCAP cells were isolated from the apical papilla of immature permanent teeth extracted for orthodontic reasons. They were exposed to various concentration of TGF-ß1 with/without pretreatment and coincubation by SB431542 (ALK/Smad2/3 inhibitor), or U0126 (MEK/ERK inhibitor). MTT assay, Western blotting and enzyme-linked immunosorbent assay (ELISA) were used to detect their effects on cell viability, and the protein expression of plasminogen activator inhibitor-1 (PAI-1), urokinase-type plasminogen activator (uPA), uPA receptor (uPAR) and their secretion. The paired Student's t-test was used for statistical analysis. RESULTS: TGF-ß1 significantly stimulated PAI-1 and soluble uPAR (suPAR) secretion of SCAP cells (P < 0.05), whereas uPA secretion was inhibited. Accordingly, TGF-ß1 induced both PAI-1 and uPAR protein expression of SCAP cells. SB431542 (an ALK5/Smad2/3 inhibitor) pretreatment and coincubation prevented the TGF-ß1-induced PAI-1 and uPAR of SCAP. U0126 attenuated the TGF-ß1-induced expression/secretion of uPAR, but not PAI-1 in SCAP. SB431542 reversed the TGF-ß1-induced decline of uPA. CONCLUSIONS: TGF-ß1 may affect the repair/regeneration activities of SCAP via differential increase or decrease of PAI-1, uPA and uPAR. These effects induced by TGF-ß1 are associated with ALK5/Smad2/3 and MEK/ERK activation. Elucidation the signalling pathways and effects of TGF-ß1 is useful for treatment of immature teeth with open apex by revascularization/revitalization procedures and tissue repair/regeneration.

Immunological analysis in paediatric HIV patients at different stages of the disease.

There are only few clinical studies on complement in well-defined (or characterized) paediatric HIV patients. Aim of this study was to evaluate the complement system and immunoglobulins in HIV-infected children and to correlate data to stage of disease. Blood samples of 127 HIV-infected children (11-134 months; 62 male : 65 female) were collected in order to evaluate humoral immunity. The patients were classified according to CDC clinical (N-asymptomatic; A-mild symptoms such as common recurrent infections; B-moderate symptoms such as Candidiasis and herpes infections, meningitis, sepsis and anaemia; C-severe symptoms such as opportunistic infections and neoplasia) and with respect to immunological criteria (T CD4(+) cell count). Analysis of complement system included the classical (CH50), alternative (APH50) pathway activities and plasma concentrations of mannan-binding lectin (MBL), of the C4 allotypic variants C4A and C4B. (ELISA), and of the C3 split product C3d (rocket immunoeletrophoresis). Immunodiagnosis also included CD4(+) and CD8(+) lymphocyte count and immunoglobulin concentrations. Complement activation and consumption was observed in all patients correlating with disease activity. Activated classical and alternative pathways and elevated C3d were significantly correlated with immunologic category 3. C3d levels were also significantly correlated with immunologic category 1. Undetectable CH50 and APH50 were found in two (group C) and 10 patients (n = 2, A = 2, B = 2, C = 4), respectively. Low MBL values were found in 13/127 but without correlation to disease severity. Undetectable C4B levels were observed in three patients, favouring the diagnosis of a complete deficiency. Although not related to clinical symptomatology, a strong ongoing complement activation can be observed in all stages of HIV infection. In contrast to earlier reports MBL could not be considered as a risk factor for HIV.

Molecular typing of Plasmodium falciparum from Giemsa-stained blood smears confirms nosocomial malaria transmission.

In this report, we describe the partial molecular characterisation of Plasmodium falciparum isolates obtained from two individuals who were involved in a probable case of accidental malaria transmission after admission to a hospital in the metropolitan area of São Paulo, Brazil. Molecular analysis of polymorphic stretches of the merozoite surface protein 1 and 2 genes using PCR-typing and nucleotide sequencing revealed that the two isolates were identical and that the identified msp-1 gene was different from all others published to date. Additional anamnestic data supported our findings and made all other possible routes of infection unlikely. The methodology used here is simple to perform and needs as little as one Giemsa-stained blood smear as starting material.

Avaliaçäo do sistema complemento em crianças, em diversos estádios da infecçäo pelo HIV: correlaçäo clínico-laboratorial / Evaluation of complement system in children in different phases of HIV infection: clinical and laboratorial correlation

Objetivo: A ativaçäo do complemento foi demonstrada em pacientes adultos infectados pelo HIV, no entanto, poucas informaçöes estäo disponíveis em crianças infectadas no período perinatal. O objetivo do presente estudo foi analisar a funçäo do sistema complemento em crianças infectads pelo HIV. Método: 127 crianças infectads pelo HIV no período neonatal (onze a 134 meses, 62 do sexo masculino: 65 do sexo feminino) foram incluídas e classificadas de acordo com os critérios clínicos e imunológicos do Centro de Controle de Doenças (Atlanta, EUA), de 1994. O diagnóstico da ativaçäo do sistema complemento foi realizado pelos seguintes ensaios: CH50 para via clássica, APH50 para via alternativa (APH50), ELISA para via das lectinas (MBL) e 'rocket' imunoeletroforese para o produto de C3,C3dg/C3d. Resultados: As infecçöes mais freqüentes foram: pneumonia bacteriana, otite, diarréia e infecçöes oportunistas como pneumonia por Pneumocystis carinii e tuberculose (31,5 por cento). A miocardiopatia foi a única apresentaçäo clínica que se relacionou com o estado imunológico (categoria 3). Nenhuma diferença estatisticamente significante nas funçöes da via clássica e alternativa foi observada entre os pacientes das diferentes categorias. Valores médios aumentados de CH50, APH50 e MBL foram verificados em pacientes nos estádios mais avançados da doença. Níveis elevados de C3d na maioria dos pacientes indicam que o complemento encontra-se ativado durante a infecçäo pelo HIV em crianças. CH50, APH50, e MBL estavam abaixo dos limites inferiores em duas, dez e duas crianças, respectivamente.

Vaginal intraepithelial neoplasia: diagnosis and management.

A review of 42 patients with vaginal intraepithelial neoplasia (VAIN) seen in Gynecologic Department of Veterans General Hospital-Taipei between 1971 and 1987 was made. The patients were from 36 to 79 years of age with a mean 58 of years. Fifteen patients (36%) had synchronous cervical neoplasia, 26 cases (62%) had antecedent cervical neoplasia and one had undergone operation for uterine myoma. Twenty-four of 27 patients with VAIN alone presented an abnormal Papanicolou smear. The upper one third of the vagina was the most common site of the lesion. Multifocal lesions were demonstrated in 58% cases while unifocal disease accounted for 42% cases who had VAIN subsequent to cervical neoplasia. Colposcopically directed biopsies were used for diagnosis in 13 patients. White epithelium and punctation were the most common findings of VAIN. While the results of different treatments were reviewed, surgical excision was found to be the most frequently employed method and radiotherapy was the second. Both were effective and 5 patients died of vaginal disease. On account of the relatively benign nature of VAIN than that of invasive vaginal cancer and the need for preservation of a functional vagina, more conservative approaches, for example, topical 5-Fluorouracil application and carbon dioxide laser vaporization, are discussed.