Diogo André Pilger

Detection of human bocavirus and human metapneumovirus by real-time PCR from patients with respiratory symptoms in Southern Brazil

The introduction of newer molecular methods has led to the discovery of new respiratory viruses, such as human metapneumovirus (hMPV) and human bocavirus (hBoV), in respiratory tract specimens. We have studied the occurrence of hMPV and hBoV in the Porto Alegre (PA) metropolitan area, one of the southernmost cities of Brazil, evaluating children with suspected lower respiratory tract infection from May 2007-June 2008. A real-time polymerase chain reaction method was used for amplification and detection of hMPV and hBoV and to evaluate coinfections with respiratory syncytial virus (RSV), influenza A and B, parainfluenza 1, 2 and 3, human rhinovirus and human adenovirus. Of the 455 nasopharyngeal aspirates tested, hMPV was detected in 14.5% of samples and hBoV in 13.2%. A unique causative viral agent was identified in 46.2% samples and the coinfection rate was 43.7%. For hBoV, 98.3% of all positive samples were from patients with mixed infections. Similarly, 84.8% of all hMPV-positive results were also observed in mixed infections. Both hBoV and hMPV usually appeared with RSV. In summary, this is the first confirmation that hMPV and hBoV circulate in PA; this provides evidence of frequent involvement of both viruses in children with clinical signs of acute viral respiratory tract infection, although they mainly appeared as coinfection agents.

Tendências na utilização de antimicrobianos em um hospital universitário, 1990-1996

OBJETIVO: Uma das principais preocupacoes mundiais quanto ao uso de medicamentos esta relacionada a utilizacao de antimicrobianos. Nos paises em desenvolvimento, poucos recursos sao empregados na monitorizacao de acoes sobre o uso racional de medicamentos. Sao, tambem, limitados os dados sobre o uso daqueles agentes em hospitais. Assim realizou-se estudo para determinar os padroes de uso de antimicrobianos em hospital num periodo de sete anos (1990 a 1996). METODOS: O estudo foi realizado em um hospital universitario, terciario, com 690 leitos, localizado em Porto Alegre, RS. Os registros hospitalares foram revisados visando identificar o consumo de antimicrobianos por pacientes internados, sendo os resultados expressos em dose diaria definida (DDD) por 100 leitos-dia. A analise de conglomerados foi realizada para determinar as tendencias de consumo dos agentes individuais. RESULTADOS: O consumo de antimicrobianos aumentou com o correr dos anos, passando de 83,8 DDD por 100 leitos-dia, em 1990, a 124,58 DDD por 100 leitos-dia em 1996. O grupo de medicamentos mais utilizado foi de penicilinas (39,6%), seguido por cefalosporinas (15,0%), aminoglicosideos (14,4%), sulfonamidas (12,8%), glicopeptideos (3,6%) e lincosaminas (3,1%). Estes grupos foram responsaveis por cerca de 90% do consumo. A analise de conglomerados do uso de antimicrobianos mostrou 13 grupos principais de tendencia de consumo. CONCLUSOES: O consumo de antimicrobianos cresceu no periodo avaliado, sendo expressivamente mais alto em comparacao com o relatado em outros estudos. Quando novas alternativas terapeuticas foram disponibilizadas no hospital, o uso de medicamentos antigos decresceu e, em alguns casos, existiu manutencao dos niveis de consumo. Quando foi realizada intervencao especifica como uma campanha para o uso correto de cefoxitina, as mudancas esperadas ocorreram.

Effect of a 36-month Pharmaceutical Care Program on the Coronary Heart Disease Risk in Elderly Diabetic and Hypertensive Patients

PURPOSE To examine the effect of a pharmaceutical care program on the coronary heart disease risk in elderly diabetic and hypertensive patients. METHODS A total of 200 elderly (> 60 years) diabetic and/or hypertensive patients were recruited into a randomized, controlled, prospective clinical trial with a 36-month follow-up, developed in a public primary health care unit in a municipality in the Brazilian State of Sao Paulo. A range of clinical measurements were evaluated at the baseline and up to 36 months afterwards. The intervention group patients received pharmaceutical care from a clinical pharmacist, whereas the control group patients received their usual care from the medical and nursing staff. The Framingham scoring method was used to estimate changes in the 10-year coronary heart disease risk scores of all the patients. RESULTS A total of 194 patients completed the study. Significant reductions (p < 0.05) in the mean values (baseline vs. 36 months) for the systolic blood pressure [156.7 mmHg vs 133.7 mmHg; P < 0.001), diastolic blood pressure (106.6 mmHg vs. 91.6 mmHg; P < 0.001),fasting glucose (135.1 mg/dL vs. 107.9 mg/dL; P < 0.001), hemoglobin A1C (7.7% vs. 7.0%; P <0.001), triglycerides (206.0 mg/dL vs. 152.5 mg/dL; P < 0.001), low-density lipoprotein (LDL)cholesterol (112.4 mg/dL vs. 102.0 mg/dL; P < 0.001), high-density lipoprotein cholesterol (55.5 mg/dL vs. 65.5 mg/dL; P < 0.001), total cholesterol (202.5 mg/dL vs. 185.9 mg/dL; P < 0.001), body mass index (26.2 kg/m2 vs. 26.1 kg/m2; P < 0.001), and abdominal circumference (103.2 cm vs. 102.5 cm; P= 0.001) were observed in the intervention group, whereas no significant changes were verified in the control group. The mean Framingham risk prediction score in the intervention group was 6.8% at baseline and decreased to 4.5%; P < 0.001) after 36 months, but remained unchanged in the control group. CONCLUSION The pharmaceutical care program resulted in better clinical measurements and reduced the cardiovascular risk scores in elderly diabetic and hypertensive patients over a 36-month period.

Diagnóstico laboratorial das leucemias mielóides agudas

Acute lymphoid leukemia (ALL) is the commonest type of cancer in childhood, representing 70% of cases. In adults its incidence is 20% only. The initial diagnosis approach consists of a peripheral blood and bone marrow cytomorphological examination. The rate of correctly classified cases is increased up to 99% through immunophenotypic study, which allows to identify the cell line (T or B) and the different stages of cell maturation. Nearly 20% derive from T cells, 75% from B-cell precursors and 5% from mature B-cells. The cytogenetic techniques have markedly contributed to the better understanding of molecular biology and ALL treatment. Chromosomal abnormalities, associated with the immunophenotyping panel, are the most important parameter for leukemia classification. Together with clinical and in vitro factors, this parameter allows the patients stratification in different risk groups, which is of great importance for establishing a prognosis and a suitable treatment. The aim of this study was to perform a bibliographic review of the laboratory methods by morphological, cytochemical, immune, cytogenetic and molecular genetic patterns, which are useful tools for acute lymphoid leukemia classification and diagnosis.

Early hematological and immunological alterations in gasoline station attendants exposed to benzene

INTRODUCTION Elucidation of effective biomarkers may provide tools for the early detection of biological alterations caused by benzene exposure and may contribute to the reduction of occupational diseases. This study aimed to assess early alterations on hematological and immunological systems of workers exposed to benzene. METHODS Sixty gasoline station attendants (GSA group) and 28 control subjects were evaluated. Environmental and biological monitoring of benzene exposure was performed in blood and urine. The potential effect biomarkers evaluated were δ-aminolevulinate dehydratase (ALA-D) activity, CD80 and CD86 expression in lymphocytes and monocytes, and serum interleukin-8 (IL-8). The influence of confounding factors and toluene co-exposure were considered. RESULTS Although exposures were below ACGIH (American Conference of Governmental Industrial Hygienists) limits, reduced ALA-D activity, decreased CD80 and CD86 expression in monocytes and increased IL-8 levels were found in the GSA group compared to the control subjects. Furthermore, according to multiple linear regression analysis, benzene exposure was associated to a decrease in CD80 and CD86 expression in monocytes. CONCLUSIONS These findings suggest, for the first time, a potential effect of benzene exposure on ALA-D activity, CD80 and CD86 expression, IL-8 levels, which could be suggested as potential markers for the early detection of benzene-induced alterations.

Differential Expression and Enzymatic Activity of DPPIV/CD26 Affects Migration Ability of Cervical Carcinoma Cells.

Dipeptidyl peptidase IV (DPPIV/CD26) is a transmembrane glycoprotein that inactivates or degrades some bioactive peptides and chemokines. For this reason, it regulates cell proliferation, migration and adhesion, showing its role in cancer processes. This enzyme is found mainly anchored onto the cell membrane, although it also has a soluble form, an enzymatically active isoform. In the present study, we investigated DPPIV/CD26 activity and expression in cervical cancer cell lines (SiHa, HeLa and C33A) and non-tumorigenic HaCaT cells. The effect of the DPPIV/CD26 inhibitor (sitagliptin phosphate) on cell migration and adhesion was also evaluated. Cervical cancer cells and keratinocytes exhibited DPPIV/CD26 enzymatic activity both membrane-bound and in soluble form. DPPIV/CD26 expression was observed in HaCaT, SiHa and C33A, while in HeLa cells it was almost undetectable. We observed higher migratory capacity of HeLa, when compared to SiHa. But in the presence of sitagliptin SiHa showed an increase in migration, indicating that, at least in part, cell migration is regulated by DPPIV/CD26 activity. Furthermore, in the presence of sitagliptin phosphate, SiHa and HeLa cells exhibited a significant reduction in adhesion. However this mechanism seems to be mediated independent of DPPIV/CD26. This study demonstrates, for the first time, the activity and expression of DPPIV/CD26 in cervical cancer cells and the effect of sitagliptin phosphate on cell migration and adhesion.

Clinical and epidemiologic profile of lower respiratory tract infections associated with human bocavirus.

Acute viral bronchiolitis (AVB) remains the leading cause of lower respiratory tract infection (LRTI) in infants under 2 years of age. Advances in molecular methods for virus detection have led to the identification of new infectious agents implicated in the development of AVB, such as human bocavirus (HBoV).

Comparação das técnicas de captura de híbridos e PCR para a detecção de HPV em amostras clínicas

INTRODUCAO E OBJETIVOS: Sao conhecidos mais de 100 tipos de papilomavirus humano (HPV), dos quais 30 tem sido reportados em infeccoes anogenitais. A infeccao tem importância clinica, pois alguns tipos virais estao associados a lesoes que podem progredir para o câncer cervical. Sabe-se que os metodos moleculares sao muito importantes para o diagnostico dessa infeccao. O objetivo do estudo e comparar a deteccao de HPV de alto risco pelo metodo de captura hibrida 2 (CH2) com a deteccao do virus pela reacao em cadeia da polimerase convencional (PCRc) e em tempo real (PCR-TR). METODOLOGIA: Foram analisadas 56 amostras ectocervicais por CH2 e, apos, por PCRc e PCR-TR. RESULTADOS: Ambas, PCRc e PCR-TR, apresentaram alta concordância entre si (95,1%), enquanto a comparacao entre as PCRs e a CH2 mostrou concordância razoavel entre os resultados (PCRc = 90,2% e PCR-TR = 87,8%). DISCUSSAO E CONCLUSAO: A CH e aceita para a deteccao do HPV, entretanto pode ser menos sensivel em comparacao com as tecnicas de PCR. A PCR-TR tem a vantagem sobre a PCRc em termos de velocidade, sendo tambem um pouco mais sensivel. Devido a alta sensibilidade e a rapidez, os metodos de PCR poderiam ser usados para a triagem de HPV em amostras ectocervicais.

Development and validity of a method for the evaluation of printed education material

Objectives To develop and study the validity of an instrument for evaluation of Printed Education Materials (PEM); to evaluate the use of acceptability indices; to identify possible influences of professional aspects. Methods An instrument for PEM evaluation was developed which included tree steps: domain identification, item generation and instrument design. A reading to easy PEM was developed for education of patient with systemic hypertension and its treatment with hydrochlorothiazide. Construct validity was measured based on previously established errors purposively introduced into the PEM, which served as extreme groups. An acceptability index was applied taking into account the rate of professionals who should approve each item. Participants were 10 physicians (9 men) and 5 nurses (all women). Results Many professionals identified intentional errors of crude character. Few participants identified errors that needed more careful evaluation, and no one detected the intentional error that required literature analysis. Physicians considered as acceptable 95.8% of the items of the PEM, and nurses 29.2%. The differences between the scoring were statistically significant in 27% of the items. In the overall evaluation, 66.6% were considered as acceptable. The analysis of each item revealed a behavioral pattern for each professional group. Conclusions The use of instruments for evaluation of printed education materials is required and may improve the quality of the PEM available for the patients. Not always are the acceptability indices totally correct or represent high quality of information. The professional experience, the practice pattern, and perhaps the gendre of the reviewers may influence their evaluation. An analysis of the PEM by professionals in communication, in drug information, and patients should be carried out to improve the quality of the proposed material.

Association of human papillomavirus and Chlamydia trachomatis with intraepithelial alterations in cervix samples

The influence of different infectious agents and their association with human papillomavirus (HPV) in cervical carcinogenesis have not been completely elucidated. This study describes the association between cytological changes in cervical epithelium and the detection of the most relevant aetiological agents of sexually transmitted diseases. Samples collected from 169 patients were evaluated by conventional cytology followed by molecular analysis to detect HPV DNA, Chlamydia trachomatis, herpes simplex virus 1 and 2,Neisseria gonorrhoeae, Mycoplasma genitalium, Trichomonas vaginalis, andTreponema pallidum, besides genotyping for most common high-risk HPV. An association between cytological lesions and different behavioural habits such as smoking and sedentariness was observed. Intraepithelial lesions were also associated with HPV and C. trachomatis detection. An association was also found between both simple and multiple genotype infection and cytological changes. The investigation of HPV and C. trachomatisproved its importance and may be considered in the future for including in screening programs, since these factors are linked to the early diagnosis of patients with precursor lesions of cervical cancer.