Effects of statins on matrix metalloproteinases and their endogenous inhibitors in human endothelial cells

Statins exert anti-inflammatory effects and downregulate matrix metalloproteinases (MMPs) expression, thus contributing to restore cardiovascular homeostasis in cardiovascular diseases. We aimed at comparing the effects of different statins (simvastatin, atorvastatin, and pravastatin) on MMP-2, MMP-9, tissue inhibitors of metalloproteinases (TIMP)-1, TIMP-2, and MMP-9/TIMP-1 and MMP-2/TIMP-2 ratios released by human umbilical vein endothelial cells (HUVEC) stimulated by phorbol myristate acetate (PMA). HUVECs were incubated with statins (0.1-10 mu M) for 12 h before stimulation with PMA 100 nM. Monolayers were used to perform cell viability assays and the supernatants were collected to determine MMPs and TIMPs levels by gelatin zymography and/or enzyme immunoassay. While treatment with PMA increased MMP-9 and TIMP-1 levels (by 556% and 159%, respectively; both P < 0.05), it exerted no effects on MMP-2 and TIMP-2 levels. Simvastatin and atorvastatin, but not pravastatin, attenuated PMA-induced increases in MMP-9 levels (P < 0.05). Only atorvastatin decreased baseline MMP-2 levels significantly (P < 0.05). We found no effects on TIMP-2 levels. Simvastatin and atorvastatin, but not pravastatin, decreased MMP-9/TIMP-1 ratio significantly (both P < 0.05), whereas atorvastatin and pravastatin, but not simvastatin, decreased MMP-2/TIMP-2 ratio significantly (both P < 0.05). Our data support the notion that statins with different physicochemical features exert variable effects on MMP/TIMP ratios (which reflect net MMP activity). Our results suggest that more lipophilic statins (simvastatin and atorvastatin), but not the hydrophilic statin pravastatin, downregulate net MMP-9 activity. However, atorvastatin and pravastatin may downregulate net MMP-2 activity. The clinical implications of the present findings deserve further investigation.

Avaliação de dois métodos laboratoriais para diagnóstico de hidatidose

A hidatidose, vulgarmente conhecida por quisto hidático, é causada pelos estados larvares do parasita Echinococcus granulosus. O seu diagnóstico baseia-se na clínica, na epidemiologia e nas técnicas imagiológicas, sendo suportado por testes serológicos. O tratamento mais comum é o cirúrgico, sendo importante o diagnóstico definitivo da doença antes da cirurgia, para se prevenir a disseminação dos quistos que pode ocorrer durante a mesma. Neste trabalho comparam-se dois métodos imunoserológicos para o diagnóstico da hidatidose: Enzyme-linked Immunosorbent Assay (ELISA) para pesquisa de Imunoglobulina G; e Fluoro-enzyme Immunoassay (FEIA) para pesquisa de Imunoglobulina E. Dos 55 indivíduos incluídos no estudo, todos em fase pré-operatória, 31% possuíam quisto hidático calcificado, 54,5% quisto hidático não calcificado e 14,5% não possuíam hidatidose, mas quistos simples. Os testes apresentaram a mesma especificidade (87,5%), sendo a sensibilidade do ELISA IgG mais baixa (63,8%) do que a do FEIA IgE (76,6%). Foi demonstrada uma boa correlação entre os dois métodos (r = 0,726, p < 0,05).

The presence of antibodies for hepatitis a virus in amazonia Didelphis marsupialis (Vertebrata, Marsupialia)

Anti-HAV was detected by enzyme - immunoassay in sera collected from 6 (18,75%) of 32 Didelphis marsupialis trapped in the Amazon region. No anti-HAV were found in the sera from 136 other wild animals, including small rodents, reptiles and other marsupials.

Seroepidemiology of hepatitis a and b in two urban communities of Rio de Janeiro, Brazil

Hepatitis B markers were determined in 397 individuals from Niterói and 680 from Nova Iguaçu and prevalences of 9.1% (1.0% of HBsAg and 8.1% of anti HBs) and 11.1% (1.8% of HBsAg and 9.3% of antiHBs) were found, respectively. The comparative prevalence of both markers in relation to age showed a higher prevalence of HBsAg in the group 21-50 years old. Considering the antiHBs antibody, it was demostrated a gradual increase with age, reaching 14.9% in Niterói and 29.1% in Nova Iguaçu in individuals over 51 years old. For hepatitis A, in 259 samples from Niterói, equally distributed by age groups, an overall prevalence of 74.5% of anti-HAV antibodies was found. This prevalence increases gradually reaching 90.0% at age over thirty. In 254 samples from Nova Iguaçu analysed, a prevalence of 90.5% of antibodies was encountered when the same criteria of distribution of samples were used. This level of prevalence reached 90.0% already in the age over ten years old. The tests were performed by enzyme immunoassay with reagents prepared in our laboratory.

Relationship between the prevalence of antibodies to hepatitis B core antigen and arbovirus in fishermen from the Ribeira Valley, Brazil

Sera from 299 fishermen 16 to 80 years old, residents in Cananeia and Iguape counties, southern cost of São Paulo State, Brazil, were studied in order to identify a possible association between the prevalence of specific antibodies to the hepatitis B virus (HBV) and exposure to haematophagus mosquitoes evaluated by the prevalence of arbovirus antibodies. This professional group presented the highest prevalence of arbovirus antibodies (54.1%) in past investigations carried out in this heavily forested region. Detection of antibody to hepatitis B core antigen (anti-HBc) in the sera was done by enzyme immunoassay (Roche). Prevalence of anti-HBc antibodies in this group was 31.4% (94/299) which is very high compared with 7.2% to 15.0% for different groups of healthy adults in State of São Paulo. No significant difference is observed between the prevalences of HBV antibodies in Iguape and Cananeia. Prevalence of anti-HBc and anti-arbovirus antibodies increases with age. There is a concordance in the distribution according to age groups of the frequency of anti-HBc and anti-arbovirus positive sera. Ag HBs was detected in 4% of the studied sera. These results support the hypothesis that the transmission of the hepatitis B virus and the arboviruses may be due to the same factor, one of the possibilities would be by anthropophilic mosquitoes.

Stool examination and rectal biopsy in the diagnosis and evaluation of therapy of schistosomiasis mansoni

From each of a group of 217 adult males selected through enzyme-immunoassay or skin-test (Group A), six stool samples were examined by both the Lutz/Hoffman, Pons & Janer (Lutz/HPJ) and Kato/Katz methods. In addition, one oogram of the rectal mucosa was performed. By these methods, schistosomiasis was detected in 44.7%, 47.5% and 40.1% of the individuals respectively. To evaluate the methods in the assessment of cure, the last 40 patients from group A, treated with a single oral dose of oxamniquine at 15 mg/kg were followed up for six months (Group B). The criteria for parasitological cure included three stool examinations by Kato/Katz and Lutz/HPJ methods, one, three and six months post-treatment and a rectal biopsy between the fourth and sixth months post-treatment. The examinations were negative in 87.5%, 90% and 95% of the patients, respectively. The efficacy of oxamniquine was 82.5% when the three methods were considered together and there was no statistically significant difference between the sensitivity of the individual methods.

Immunodiagnosis of hydatid disease: evaluation of antigens from hydatid cyst fluid and the vesicularfluid of Taenia crassiceps metacestode

The specificity and sensitivity of the enzyme immunoassay (EIA), presently used in South America areas where hydatidosis caused by Echinococcus granulosus is endemic, was compared to two alternative EIA. One of these uses an hydatid antigen of different prepraration and the other vesicular fluid of Taenia crassiceps cisticerci (VFCC). The effect of previous neutralization in the serum sample of antibodies anti-normal ovine or murine sera and anti-phosphorylcholine on the diagnostic efficiency of these EIA was studied. The frequency of distribution of the titers obtained with normal sera, hydatid sera positive to DD5 test and hydatid sera negative to DD5 test in three EIA systems was analyzed. Results showed a significant decrease of sensitivity of the EIA using VFCC when compared to these EIA using hydatid antigens. This makes inconvenient the use of VFCC for the immunodiagnosis of hydatid disease. No significant differences between the two EIA using hydatid antigens were observed. SDS-PAGE analysis showed remarkable differences between the VFCC and the hydatid antigens composition and some differences among these latters probably due to manufacturing procedures.

Human parvovirus B19 infection: clinical and epidemiological study of 24 cases

From March 1994 to November 1995 24 cases of human parvovirus B19 infection were seen at the Infectious Diseases Department of the Hospital Universitário Antônio Pedro, Niterói - RJ. Serum samples for IgM detection (capture enzyme immunoassay) were positive from the 1st to the 27th day after the onset of the exathema. The classical features of erythema infectiosum (slapped cheecked syndrome) were observed in 8 (33.3%) cases all of them children. Eight patients (6 adults and 2 children) presented a symmetrical polyartropathy, seen more frequently in women. These results show that B19 infection diagnosis is difficult when the disease does not present the classical features and because of the frequent involvement of the joints this infection should be considered in the differential diagnosis of early rheumatoid arthritis.

Seroprevalence of toxoplasmosis in Fortaleza, Ceará, Brazil

A serological survey of Toxoplasma gondii infection in population groups in Fortaleza, Brazil, was conducted, in order to identify seroprevalence rates according to age and risk factors associated with acquired infection. A Toxoplasma IgG-antibody enzyme immunoassay (Sanofi Pasteur Diagnostics, Marnes la Coquette, France) was employed to assess the immunity. Public day-care centers and schools were randomly selected, while three large antenatal clinics and maternity units were choosen by its importance. Population groups and results of 997 blood tests were: 227 children (mean age 3.8 years), 22.8% seropositives; 584 students (mean 11.4 years), 58.4%, and pregnant and postpartum women (mean 24 years), 71.5% seropositives (p < 0.001). Of 256 participants reporting close contact with cats, 59.8% were seropositive, in contrast with 51% seropositives without contact (relative risk 1.17; 95% confidence interval 1.04-1.33; p = 0.01). Having three or more siblings at home was related to higher seroprevalence in children and students (relative risk 1.39; 95% confidence interval 1.21-1.60; p < 0.01). In conclusion, toxoplasmosis seroprevalence showed a rapid increase during the first ten years of life, in association with close contact with cats and larger households, probably related to inappropriate hygiene and child-care practices.

Seasonal variation of hepatitis A virus infection in the city of Rio de Janeiro, Brazil

Hepatitis A virus (HAV) infection constitutes a major public health problem in Brazil. The transmission of HAV is primarily by fecal-oral route so the water is an important vehicle of HAV dissemination. There is a great incidence of acute cases of hepatitis A in some areas of Brazil however the seasonal variation of these cases was not documented. The aim of this study was to determine the seasonality of HAV infection in Rio de Janeiro. From January 1999 to December 2001, 1731 blood samples were collected at the National Reference Center for Hepatitis Viruses in Brazil (NRCHV). These samples were tested by a commercial enzyme-immunoassay to detect anti-HAV IgM antibodies. Yearly positive rates were 33.74% in 1999, 32.19% in 2000, and 30.63% in 2001. A seasonal variation was recognized with the highest incidence in spring and summer. Furthermore a seasonal increase in incidence of HAV infection was found during the rainy season (December to March) because the index of rains is very high. It is concluded that HAV infections occur all year round with a peak during hot seasons with great number of rains.

Prevalence of IgG antibodies specific to Toxoplasma gondii among blood donors in Recife, Northeast Brazil

A serological survey of Toxoplasma gondii infection in blood donors was carried out in order to identify seroprevalence in Recife, Brazil. Sera from 160 individuals (119 male and 41 female) were evaluated by using a Toxoplasma IgG-antibody enzyme immunoassay (Denka Seiken Co., LTD., Tokyo, Japan). The seropositive percentual for males (79.0%) showed to be higher (p < 0.05) than for females (63.4%). This percentage increases with age, ranging from 18.2% to 92.6% for individuals aging under 20 and 40-50 years old, respectively. For women of childbearing age (18-40 years) it was found a prevalence of 51.6%.

Childhood astrovirus-associated diarrhea in the ambulatory setting in a Public Hospital in Cordoba city, Argentina

Human astroviruses have been increasingly identified as important agents of diarrheal disease in children. However, the disease burden of astrovirus infection is still incompletely assessed. This paper reports results on the epidemiological and clinical characteristics of astrovirus-associated diarrhea, as well as the impact of astrovirus infection on the ambulatory setting at a Public Hospital in Córdoba city, Argentina. From February 2001 through January 2002, 97 randomly selected outpatient visits for diarrhea among children < 36 months old were enrolled. A single specimen of stool from each child was collected and tested for astrovirus antigen by enzyme immunoassay. Astroviruses were detected in 12.37% of the diarrheal episodes. All the positive cases occurred in children 4 to 18 months, but the highest rate was in children aged 4 to 6 months (23.80%). The clinical symptoms of astrovirus associated-diarrhea were fever 41.66%, vomiting 25.00% and dehydration 8.33%; overall 16.66% required hospitalization. Astrovirus was identified through the year and no seasonally pattern was detected (cool semester 15.21% versus warm semester 9.80% p > 0.05). According to our estimation about one out of seventy-four children in this cohort would be assisted annually for an astroviral-diarrheal episode in the Public Hospital and one out of eight diarrheal cases could be attributed to astrovirus infection. Astrovirus is a common symptomatic infection in pediatric outpatient visits in the public hospital in the study area, contributing 12.37% of the overall morbidity from diarrhea.

Hospitalização em Crianças com Infecção por Rotavírus

Introdução. A infecção por rotavírus é a principal causa de diarreia aguda em todo o mundo. Nos países desenvolvidos não constitui uma causa importante de morte, mas cursa com uma alta morbilidade. Objectivo. Caracterizar a infecção por rotavírus em crianças hospitalizadas em dois Departamentos de Pediatria de Lisboa. Material e métodos. Revisão casuística dos internamentos com o diagnóstico de infecção por rotavírus, num hospital central especializado e num hospital geral na Zona Metropolitana de Lisboa, entre Janeiro e Dezembro de 2005. O diagnóstico foi efectuado através da identificação de antigénios virais nas fezes por “enzyme immunoassay”. Resultados. Foram analisados 92 casos; 82% ocorreram entre Dezembro e Março e 52,8% em crianças entre os três e os doze meses. Em metade dos casos registou-se bom nível socio-económico. Os factores de risco epidemiológicos encontrados foram: frequência de instituição de ensino ou ama em 21/38 (55%), contacto com pessoas com sintomatologia semelhante em 10/53 (19%) e irmãos com idade inferior a cinco anos em 25/76 (33%) das crianças. As infecções nosocomiais foram responsáveis por 26% dos casos estudados. A clínica cursou com: diarreia aquosa (96%), vómitos (87%) e febre (69%). Ocorreram complicações em 19/92 (21%) crianças e estas foram mais frequentes em lactentes com menos de seis meses de idade (35% vs. 16%, p=0,058). A mediana da duração de internamento foi cinco dias e o custo hospitalar directo variou entre 629,63 e 2342,38 euros. Discussão. O número de internamentos por infecção por rotavírus, especialmente em lactentes, a frequência de infecções nosocomiais por este agente, as complicações inerentes e os elevados custos, reflectem a importância da infecção por este agente em países desenvolvidos como Portugal.

Performances of HTLV serological tests in diagnosing HTLV infection in high-risk population of São Paulo, Brazil

Testing problems in diagnosing human T-lymphotropic virus (HTLV) infection, mostly HTLV-II, have been documented in HIV/AIDS patients. Since December 1998, the Immunology Department of Instituto Adolfo Lutz (IAL) offers HTLV-I/II serology to Public Health Units that attend HTLV high-risk individuals. Two thousand, three hundred and twelve serum samples: 1,393 from AIDS Reference Centers (Group I), and 919 from HTLV out-patient clinics (Group II) were sent to IAL for HTLV-I/II antibodies detection. The majority of them were screened by two enzyme immunoassays (EIAs), and confirmed by Western Blot (WB 2.4, Genelabs). Seven different EIA kits were employed during the period, and according to WB results, the best performance was obtained by EIAs that contain HTLV-I and HTLV-II viral lysates and rgp21 as antigens. Neither 1st and 2nd, nor 3rd generation EIA kits were 100% sensitive in detecting truly HTLV-I/II reactive samples. HTLV-I and HTLV-II prevalence rates of 3.3% and 2.5% were detected in Group I, and of 9.6% and 3.6% in Group II, respectively. High percentages of HTLV-seroindeterminate WB sera were detected in both Groups. The algorithm testing to be employed in HTLV high-risk population from São Paulo, Brazil, needs the use of two EIA kits of different formats and compounds as screening, and because of high seroindeterminate WB, may be another confirmatory assay.

Canine visceral leishmaniasis: study of methods for the detection of IgG in serum and eluate samples

The Brazilian Ministry of Health recommends the culling and euthanasia of dogs with a positive serological test for canine visceral leishmaniasis (CVL). In the Municipality of Rio de Janeiro, the technique used for the diagnosis of CVL is the indirect fluorescent antibody test (IFAT), using blood samples eluted on filter paper (eluate). A dog survey was conducted over a period of one year in the region of Carapiá, in order to evaluate the diagnosis of CVL in this region. All animals underwent clinical examination, and blood samples (serum and eluate) were collected for analysis by enzyme immunoassay (ELISA) and IFAT. A skin biopsy was obtained for parasitological examination (culture). A total of 305 animals were studied and Leishmania chagasi was isolated from nine animals. Sensitivity and specificity were 100% and 96.6% for ELISA, respectively, 100% and 65.5% for IFAT (cut-off at a 1:40 dilution), 100% and 83.4% for IFAT (cut-off at a 1:80 dilution), and 22.2% and 97.0% for eluate IFAT. In conclusion, ELISA was the best tool for the diagnosis of CVL among the serological techniques tested. The present results suggest the need for a better evaluation of filter paper IFAT as the only diagnostic method for CVL in the Municipality of Rio de Janeiro.