Contribution of matrix vesicles and alkaline phosphatase to ectopic bone formation

Endochondral calcification involves the participation of matrix vesicles (MVs), but it remains unclear whether calcification ectopically induced by implants of demineralized bone matrix also proceeds via MVs. Ectopic bone formation was induced by implanting rat demineralized diaphyseal bone matrix into the dorsal subcutaneous tissue of Wistar rats and was examined histologically and biochemically. Budding of MVs from chondrocytes was observed to serve as nucleation sites for mineralization during induced ectopic osteogenesis, presenting a diameter with Gaussian distribution with a median of 306 ± 103 nm. While the role of tissue-nonspecific alkaline phosphatase (TNAP) during mineralization involves hydrolysis of inorganic pyrophosphate (PPi), it is unclear how the microenvironment of MV may affect the ability of TNAP to hydrolyze the variety of substrates present at sites of mineralization. We show that the implants contain high levels of TNAP capable of hydrolyzing p-nitrophenylphosphate (pNPP), ATP and PPi. The catalytic properties of glycosyl phosphatidylinositol-anchored, polidocanol-solubilized and phosphatidylinositol-specific phospholipase C-released TNAP were compared using pNPP, ATP and PPi as substrates. While the enzymatic efficiency (k cat/Km) remained comparable between polidocanol-solubilized and membrane-bound TNAP for all three substrates, the k cat/Km for the phosphatidylinositol-specific phospholipase C-solubilized enzyme increased approximately 108-, 56-, and 556-fold for pNPP, ATP and PPi, respectively, compared to the membrane-bound enzyme. Our data are consistent with the involvement of MVs during ectopic calcification and also suggest that the location of TNAP on the membrane of MVs may play a role in determining substrate selectivity in this micro-compartment.

Hepatitis B and C virus infection among Brazilian Amazon riparians

INTRODUCTION: Viral hepatitis is a major public health concern in Brazil. There are few past studies on this issue, especially among riparian communities. This study aims at determining the seroprevalence of viral hepatitis B and C in the riparian community of Pacuí Island, within the Cametá municipality of Pará State, Brazil. Moreover, this study aims to investigate the principal risk factors that this community is exposed to. METHODS: The current study has accessed blood samples from 181 volunteers who have answered an epidemiological questionnaire. Analyses on serological markers have been tested with commercial ELISA kits for detecting HBsAg, total anti-HBc, anti-HBs, and anti-HCV. Within seroreactive patients for HCV, RT-PCR and line probe assay have been performed to identify the viral genotype. RESULTS: In the serological marker analysis for hepatitis B, no reactivity for HBsAg, rate of 1.1% for total anti-HBc, and rate of 19.3% for anti-HBs have been observed. On hepatitis C, 8.8% seroprevalence has been found, in which 62.5% have gotten viral RNA. Among the risk factors studied, the following have been highlighted: non-use of condoms, sharing of cutting instruments, use of illicit drugs, and reports of family disease with HBV or HCV. CONCLUSIONS: The vaccination coverage against HBV is low, and the high prevalence of HCV within this community has been observed.

Prevalencia de hepatitis B, hepatitis C y sífilis en trabajadoras sexuales de Venezuela

OBJETIVO: En Venezuela las trabajadoras sexuales reciben un control sanitario para la sífilis y el virus de inmunodeficiencia humana (VIH). Sin embargo, otras importantes infecciones de transmisión sexual no son evaluadas. Así, se realizó este estudio con el objetivo de determinar el nivel socio-cultural de un grupo de trabajadores sexuales y su relación con la sero-presencia de marcadores de Hepatitis C y Hepatitis B, en adición a la evaluación de rutina. MÉTODOS: Se evaluaron 212 trabajadoras sexuales, que acudieron al control sanitario en el servicio de infecciones de transmisión sexual, de la ciudad de Los Teques, Venezuela. Fueron entrevistadas en cuanto a edad, nivel educativo, uso de anticonceptivos y del condón. Se les tomó una muestra de sangre para determinar sífilis, antígeno de superficie de hepatitis B (HBsAg) y la presencia de anticuerpos contra el core de hepatitis B (anti-HBc), virus de hepatitis C (anti-HC) y VIH. Los datos fueron evaluados estadísticamente por Chi-cuadrado y correlación de Pearson. RESULTADOS: La prevalencia fue de 2,4% para sífilis, 0,5% para anti-HC, 3,8% para HBsAg y 13,8% para anti-HBc. Un aumento en la prevalencia de marcadores de hepatitis B se correlacionó con un bajo nivel educativo (p<0,05) e incremento en la edad (p<0,05). No se encontró ningún caso positivo de VIH. La encuesta reveló que el 38,5% de las trabajadoras sexuales nunca utilizan el condón y un 25,6% de ellas no utiliza ningún tipo de método anticonceptivo. CONCLUSIONES: Es necesario implementar planes de inmunización de hepatitis B en este grupo de mujeres, así como campañas de educación sobre la importancia del uso del condón para disminuir la probabilidad de contraer infecciones de transmisión sexual.

Infecção pelo HIV, hepatites B e C e sífilis em moradores de rua, São Paulo

OBJETIVO: Estimar a prevalência das infecções pelo HIV, vírus das hepatites B e C, e da sífilis em moradores de rua. MÉTODOS: Estudo transversal com intervenção educativa, realizado no município de São Paulo, de 2002 a 2003. Selecionou-se amostra de conveniência de moradores de rua que utilizavam albergues noturnos, segundo os critérios: >18 anos e não apresentar distúrbios psiquiátricos. Em entrevistas, foram coletados dados sociodemográficos e de comportamento, e realizados exames laboratoriais para HIV, hepatite B e C e sífilis, e aconselhamento pós-teste. RESULTADOS: Participaram 330 usuários dos albergues, com 40,2 anos (média), 80,9% homens, nas ruas, em média, há um ano. Observaram-se prevalências de 1,8% de HIV, 8,5% de vírus de hepatite C, 30,6% de infecção pregressa por hepatite B, 3,3% de infecção aguda ou crônica pelo vírus hepatite B e 5,7% de sífilis. Uso consistente de preservativo foi referido por 21,3% e uso de droga injetável, por 3% dos entrevistados. A positividade para HIV foi de 10% e 50% para vírus da hepatite C entre usuários de drogas injetáveis, versus 1,5% para HIV e 7,3% para hepatite C nos demais, evidenciando associação entre esse vírus e uso de droga injetável. Prisão anterior foi referida por 7,9% das mulheres e 26,6% dos homens, com prevalência de 2,6% para HIV e 17,1% para vírus da hepatite C. CONCLUSÕES: As elevadas prevalências de HIV e vírus de hepatite B e C requerem programas de prevenção baseados na vacinação contra hepatite B, diagnóstico precoce dessas infecções e inserção dos moradores de rua em serviços de saúde.

Hepatites B e C em usuários de drogas injetáveis vivendo com HIV em São Paulo, Brasil

OBJETIVO: Descrever o perfil de usuários de drogas injetáveis vivendo com HIV/Aids e estimar a prevalência de hepatites B e C nesse grupo. MÉTODOS: Estudo transversal realizado com 205 pessoas vivendo com HIV/Aids, usuários de drogas injetáveis em acompanhamento em três unidades de atendimento da rede pública do Município de São Paulo, em 2003. Foi selecionada amostra não-probabilística, obtida de forma consecutiva e voluntária, nos dias em que compareciam para consulta nas unidades de atendimento. Por meio de entrevistas, foram levantados dados pessoais e informações sobre comportamento sexual, uso de drogas e conhecimento de hepatites. Foram realizados testes para detecção da infecção pelos vírus das hepatites B e C. RESULTADOS: Dos entrevistados, 81% eram homens e 19% mulheres, com idade média de 39 anos (dp=6,1) e seis anos de educação formal (dp=2,0). Não havia diferença em relação ao estado marital entre os sexos, 48% eram solteiros, 42% casados e 8% divorciados. A idade média do primeiro uso de tabaco, álcool e drogas ilícitas foi 13, 15 e 18 anos, respectivamente. Prevalências de hepatites B e C foram, respectivamente, de 55% (IC 95%: 49;63) e 83% (IC 95%: 78;88). Antes de usar droga injetável pela primeira vez, 80% dos respondentes não tinham ouvido falar de hepatites B e C. CONCLUSÕES: A alta prevalência de hepatites B e C e o baixo nível de conhecimento sobre a doença justificam a inclusão de esclarecimentos sobre as infecções hepáticas e de vacinação contra hepatite B nas estratégias de redução de danos pelo HIV.

Quimiotipagem do Cryptococcus neoformans. Revisão da literatura. Novos dados epidemiológicos sobre a criptococose. Nossa experiência com o emprego do meio de C.G.B. no estudo daquela levedura

O presente trabalho, além da revisão da literatura sobre quimiotipagem do C. neoformans, com novos dados sobre a epidemiologia da criptococose, teve por finalidade principal a caracterização das duas variedades desta levedura em pacientes com neurocriptococose, HIV + e HIV -. As variedades neoformans e gattii estão hoje bem definidas bioquimicamente, com o emprego do meio C.G.B., proposto por KWON-CHUNG et al. (1982) 24. O isolamento do C. neoformans var. gattii das flores e folhas do Eucalyptus camaldulensis e do Eucalyptus tereticornis, na Austrália, através dos trabalhos de ELLIS & PFEIFFER (1990)16 e PFEIFFER & ELLIS (1992)41, possibilitou investigações epidemiológicas das mais interessantes sobre este microrganismo, levedura capsulada a qual SANFELICE50, 51, na Itália, em 1894 e 1895 despertou a atenção do meio médico. BUSSE8, em 1894, descrevia o primeiro caso de criptococose humana sob a forma de lesão óssea, simulando sarcoma. As pesquisas nacionais sobre o assunto em foco foram destacadas, seguindo-se a experiência dos Autores com o meio de C.G.B. (L - canavanina, glicina e azul de bromotimol). Foi possível, através deste meio o estudo de 50 amostras de líquor, sendo 39 procedentes de aidéticos (78%) e 11 de não aidéticos (22%). De pacientes HIV+, 37 (74%) foram identificados como C. neoformans var. neoformans e 2 (4%) como C. neoformans var. gattii. Dos HIV- 8 ( 16%) foram classificados como C. neoformans var. neoformans e 3 (6%) como C. neoformans var. gattii. Através deste trabalho, evidencia-se a importância da neurocriptococose, principalmente entre os aidéticos, demonstrando-se mais uma vez o interesse do meio CGB na quimiotipagem do C. neoformans em suas duas variedades, ganhando em importância a demonstração de que duas espécies de eucalipto podem funcionar como "árvores-hospedeiras" para o Cryptococcus neoformans var. gattii.

PREVALENCE OF HEPATITIS B AND HEPATITIS C MARKERS IN ALCOHOLICS WITH AND WITHOUT CLINICALLY EVIDENT HEPATIC CIRRHOSIS

We assessed the frequency of serological markers of hepatitis B virus (HBV) and hepatitis C virus (HCV) infections in 365 alcoholics by determining, by ELISA, the presence of HBsAg, anti-HBc, anti-HBs and anti-HCV. Fifty patients were cirrhotics and 315 had no evidence of hepatic cirrhosis; of the latter HBsAg was assessed in all, anti-HBc and anti-HBs in 130, and anti-HCV in 210. Among the alcoholics the frequencies of HBsAg (1.9%), anti-HBc (28.3%) and anti-HCV (3.8%) were higher (p<0.001) than among the controls (N=17,059), 0.4%, 4.0% and 0.4% respectively. The frequency of positive HBsAg was higher (p<0.001) in the cirrhotic patients (8.0%) than in alcoholics without cirrhosis (0.95%) and in controls (0.4%), and similar between the latter; of anti-HBc in alcoholics without cirrhosis (28.5%) was similar in cirrhotics patients (28.0%) and higher (p<0.001) than in the controls (4.0%); of anti-HBs in alcoholics without cirrhosis (20.8%) was similar to that of the cirrhotic patients (10.0%), and the anti-HCV was similar between alcoholics with (6.0%) and without cirrhosis (3.3%) and higher (p<0.001) than in controls (0.4%). We concluded that: a) alcoholics with or without cirrhosis have similar frequencies of infection with HBV and HCV between them, and higher than in nonalcoholics; b) alcoholics without cirrhosis had a frequency of HBV active infection (HBsAg+) which was similar to the controls, whereas among those who progressed to cirrhosis this frequency was significantly higher, what suggests that HBV may be implicated in the pathogenesis of cirrhosis in a few alcoholic individuals.

Prevalence of hepatitis B and C in the sera of patients with HIV infection in São Paulo, Brazil

The objective of this study was to evaluate the prevalence of hepatitis B and C viruses in a group of HIV infected patients, followed at a single institution since 1996. 1,693 HIV positive patients (1,162 male, 531 female) were tested for HBV infection. Virological markers for HBV included HBsAg and total anti-HBc by ELISA. 1,457 patients (1,009 male, 448 female) were tested for HCV infection. Detection of HCV antibodies was carried out by ELISA. A sample of HCV antibody positive patients was tested for HCV by PCR to confirm infection. Of 1,693 patients tested for HBV, 654 (38.6%) and 96 (5.7%) were anti-HBc and HBsAg positive, respectively. Of 1,457 patients tested for HCV, 258 (17.7%) were anti-HCV positive. 82 of these patients were also tested by PCR and 81 were positive (98%). Of 1,411 patients tested for HBV and HCV 26 (1.8%) were positive for both viruses.

Hepatitis B and C virus markers among patients with hepatosplenic mansonic schistosomiasis

PURPOSE: To evaluate the frequency and the consequences of the co-infection of hepatitis B and C viruses in patients with hepatosplenic schistosomiasis (HSS). METHODS: B and C serologic markers, exposure to risk factors, biochemical assays, upper gastrointestinal endoscopies, and abdominal ultrasonograms were evaluated in 101 patients with HSS from 1994 to 1997. Whenever possible, PCR was tested and histopathological studies were reviewed. RESULTS: At least one HBV virus marker was found in 15.8%, and anti-HCV was detected in 12.9% of the subjects. The seropositive subjects tended to be older than the seronegative ones. A history of blood transfusion was significantly related to the presence of anti-HCV. Three (18.75%) out of 16 subjects exposed to B virus were HBsAg positive. Eleven (84.6%) out of thirteen patients who were anti-HCV positive demonstrated viral activity. Patients with ongoing viral infection presented a higher average level of liver aminotransferases, a higher frequency of cell decompensation and a higher rate of chronic hepatitis. Portal hypertension parameters were not influenced by viral exposure. CONCLUSIONS: The rate of hepatitis B and C viruses serologic markers observed in the patients with HSS was higher than the control group. The co-infection was responsible for a higher frequency of cell decompensation.

Comparative morphometry and morphology of Anopheles aconitus Form B and C eggs under scanning electron microscope

Comparative morphometric and morphological studies of eggs under scanning electron microscope (SEM) were undertaken in the three strains of two karyotypic forms of Anopheles aconitus, i.e., Form B (Chiang Mai and Phet Buri strains) and Form C (Chiang Mai and Mae Hong Son strains). Morphometric examination revealed the intraspecific variation with respect to the float width [36.77 ± 2.30 µm (Form C: Chiang Mai strain) = 38.49 ± 2.78 µm (Form B: Chiang Mai strain) = 39.06 ± 2.37 µm (Form B: Phet Buri strain) > 32.40 ± 3.52 µm (Form C: Mae Hong Son strain)] and number of posterior tubercles on deck [2.40 ± 0.52 (Form B: Phet Buri strain) = 2.70 ± 0.82 (Form B: Chiang Mai strain) < 3.10 ± 0.32 (Form C: Chiang Mai strain) = 3.20 ± 0.42 (Form C: Mae Hong Son strain)], whereas the surface topography of eggs among the three strains of two karyotypic forms were morphologically similar.

Serological markers and risk factors for hepatitis B and C viruses in patients infected with human immunodeficiency virus

Both hepatitis B and hepatitis C viruses (HBV and HCV) infection are common in HIV-infected individuals as a result of shared risk factors for acquisition. A serological study for HBV and HCV was performed in 251 HIV-positive individuals from Medellín, Colombia. A qualitative RT-PCR for HCV was done in 90 patients with CD4+ T-cell count < 150 per mm³. Serological markers for HBV infection were present in 97 (38.6%) patients. Thirty six of them (37.1%) had isolated anti-HBc. A multivariate analysis indicated that the following risk factors were significantly associated with the presence of these markers: age (OR = 1.05, 95% CI: 1.01-1.08), pediculosis pubis (OR = 1.83, 95% CI: 1.01-3.33), men who have sex with men and women (OR = 3.23, 95% CI: 1.46-7.13) and men who have sex only with men (OR = 3.73, 95% CI: 1.58-8.78). The same analysis restricted to women showed syphilis as the only significant risk factor. Thus, HBV infection was considerably associated with high risk sexual behavior. HCV was present in only two (0.8%) of HIV patients. Both of them were positive by RT-PCR and anti-HCV. This low frequency of HIV/HCV coinfection was probably due to the uncommon intravenous drug abuse in this population. The frequent finding of isolated anti-HBc warrants molecular approaches to rule out the presence of cryptic HBV infection.

Seroprevalence of hepatitis B and C infection markers among children and adolescents in the southern Brazilian region

Hepatitis B virus (HBV) and hepatitis C virus (HCV) infections account for a substantial proportion of liver diseases worldwide. The aim of this study was to determine the prevalence of HBV and HCV serological markers among children and adolescents and verify the epidemiology of the HBV infection over than a decade of the introduction of vaccination program. Serologic markers to HBsAg, total anti-HBc and anti-HCV had been tested in 393 samples. The seropositivity for HBsAg was 0.76% and for total anti-HBc was 1.02%. Copositivity between HBsAg and total anti-HBc was verified in 0.76% of the analyzed samples. There was no seropositivity for anti-HCV marker. The seroprevalence of HBV infection markers among children and adolescents in the southern Brazilian region is high compared to that reported in other countries. Preventive measures, such as educational activities in addition to the universal childhood HBV vaccination, should be initiated in order to reduce the morbimortality and the economic burden associated with the disease.