Alguns aspectos citológicos da Leptospira Icterohaemorrhagiae em microscópio eletrônico

Os autores estudando culturas de 11 e 12 meses de Leptospira icterohaemorrhagiae, mantidas em meio de Schüffner modificado por Verwoort, observaram em microscópio eletrônico, corpos de leptospira com zonas de condensação do citoplasma assumindo aspecto granular Figs. 1, 3, 4, 9, 7, 12. Nas Figs. 6 e 7 vemos com nitidez um dêsses grânulos desmembrando-se do corpo da leptospira e cuja imagem é semelhante a outros grânulos esparsos. O número dessas formações é variável, parecendo depender de muitas causas. A deformação do corpo da leptospira parece depender da perda de consistência da membrana em virtude de alteração da sua permeabilidade, fenomenos que deve preceder á formação dos grânulos. Notamos também nas mesmas condições de experiência, como na Fig. 3, que os referidos grânulos tem dois aspectos diferentes: um é o que já referimos de forma irregular e opaco como se fôsse uma imagem de cromatina condensada e outro aspecto é também de forma irregular, porém, transparente como se houvesse esvaziado seu conteúdo primitivo, Figs. 5, 7 e 13. Na fig. 12, observamos o esvaziamento de uma forma destas o retraimento do seu conteúdo em outra, observação facilitada com auxílio de lente. A fig. 2 revela um aspecto frqüente encontrado em trabalhos desta natureza como no de CZEKALWSKI, J. W. e EAVES, G., isto é: formação de um grânulo com posterior segmentação do mesmo. Finalmente a Fig. 11 nos mostra grânulos com a mesma densidade que os fragmentos dos corpos da leptospira. Como dissemos acima, a cultura contendo material com êste aspecto foi inoculada em cobaias cuja hemocultura nos forneceu de novo o germe reisolado, o que faz admitir constituirem êsses grânulos uma fase no ciclo vital da leptospira ou então elemento capazer de manter condições e fatôres necessários à continuidade da espécie.

Observações sôbre eliminação de dejeções e tempo de sucção em alguns triatomíneos sul-americanos

São relatadas as observações a respeito dos hábitos de defecação e de sucção em 6 espécies de hemípteros hematófagos. O quadro abaixo reúne os resultados mais gerais obtidos, apreciados em conjunto: Espécies de exemplares utilizados; % de insetos que defecaram durante ou logo após a picada; N.º médio de defecações nas 3 primeiras horas após o repasto; Duração média de sucção (minutos); % de insetos que sugaram sem interromper a picada. R. prolixus 2 machos, 8 fêmeas e 10 ninfas (50,0; 13,7; 14,2; 20,0). R. neglectus 2 machos, 8 fêmeas e 10 ninfas (30,0; 9,6; 18,5; 20,0). T. infestans 2 machos, 11 fêmeas e 20 ninfas (30,0; 7,1; 15,5; 47,5). P. megistus 6 machos, 10 fêmeas e 12 ninfas (22,7; 3,4; 22,7; 82,1). T. sordida 13 machos e 11 ninfas (12,5; 4,5; 20,0; 79,2). T. vitticeps 11 ninfas (0; 6,2; 26,8; 90,9). De modo geral, a capacidade de defecar no ato da picada mostrou-se diretamente proporcional ao número de defecações (nas três primeiras horas) e à freqüência das interrupções da picada, e inversamente proporcional ao tempo de duração da sucção. Os barbeiros adultos se mostraram mais aptos a defecar no ato da picada do que em fase de ninfa, as fêmeas mais do que os machos. Das espécies observadas, o R. prolixus foi a que melhores condições demonstrou para realizar a contaminação fecal do hospedeiro vertebrado. Sugere-se que um estudo mais aprofundado sôbre a eficácia contaminativa dos diversos transmissores do S. cruzi seja feito de preferência no verão e à noite, em zonas onde grassa endêmicamente a doença de Chagas.

Infecção experimental de Lutzomyia whitmani em cães infectados com Leishmania braziliensis braziliensis

Lutzomyia (N.) whitmani foi infectada em lesões leishmanióticas de três de nove cães parasitados por Leishmania braziliensis braziliensis . Os índices de infecção desses flebotomíneos foram 8,3% (1/12), 7,1% (1/14) e 1,8% (3,160), respectivamente. Por outro lado, 180 Lu. whitmani que se alimentaram em áreas não-ulceradas em um dos cães foram negativos, após dissecação. É discutida a potencialidade de Lu. whitmani como vector de L.B. braziliensis na região endêmica de Três Braços, Bahia, Brasil.

Identification of Human T-lymphotropic Virus Type I (HTLV-I) Subtypes Using Restrited Fragment Length Polymorphism in a Cohort of Asymptomatic Carriers and Patients with HTLV-I-associated Myelopathy/tropical Spastic Paraparesis from São Paulo, Brazil

Although human T-lymphotropic virus type I (HTLV-I) exhibits high genetic stability, as compared to other RNA viruses and particularly to human immunodeficiency virus (HIV), genotypic subtypes of this human retrovirus have been characterized in isolates from diverse geographical areas. These are currently believed not to be associated with different pathogenetic outcomes of infection. The present study aimed at characterizing genotypic subtypes of viral isolates from 70 HTLV-I-infected individuals from São Paulo, Brazil, including 42 asymptomatic carriers and 28 patients with HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP), using restricted fragment length polymorphism (RFLP) analysis of long terminal repeat (LTR) HTLV-I proviral DNA sequences. Peripheral blood mononuclear cell lysates were amplified by nested polymerase chain reaction (PCR) and amplicons submitted to enzymatic digestion using a panel of endonucleases. Among HTLV-I asymptomatic carriers, viral cosmopolitan subtypes A, B, C and E were identified in 73.8%, 7.1%, 7.1% and 12% of tested samples, respectively, whereas among HAM/TSP patients, cosmopolitan A (89.3%), cosmopolitan C (7.1%) and cosmopolitan E (3.6%) subtypes were detected. HTLV-I subtypes were not statistically significant associated with patients' clinical status. We also conclude that RFLP analysis is a suitable tool for descriptive studies on the molecular epidemiology of HTLV-I infections in our environment.

Antibacterial activity of extracts and neolignans from Piper regnellii (Miq.) C. DC. var. pallescens (C. DC.) Yunck

The evaluation of the activity of the aqueous and ethyl acetate extracts of the leaves of Piper regnellii was tested against gram-positive and gram-negative bacteria. The aqueous extractdisplayed a weak activity against Staphylococcus aureus and Bacillus subtilis with minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) of 1000 µg/ml. The ethyl acetate extract presented a good activity against S. aureus and B. subtilis with MIC and MBC at 15.62 µg/ml. In contrast to the relative low MICs for gram-positive bacteria, gram-negative bacteria were not inhibited by the extracts at concentrations < 1000 mg/ml. The ethyl acetate extract was fractionated on silica gel into nine fractions. The hexane and chloroform fractions were active against S. aureus (MIC at 3.9 µg/ml) and B. subtilis (MIC at 3.9 and 7.8 µg/ml, respectively). Using bioactivity-directed fractionation, the hexane fraction was rechromatographed to yield the antimicrobial compounds 1, 2, 5, and 6identified as eupomatenoid-6, eupomatenoid-5, eupomatenoid-3, and conocarpan, respectively. The pure compounds 1 and 2 showed a good activity against S. aureus with MIC of 1.56 µg/ml and 3.12 µg/ml, respectively. Both compounds presented MIC of 3.12 µg/ml against B. subtilis. The pure compound 6 named as conocarpan was quite active against S. aureus and B. subtilis with MIC of 6.25 µg/ml. The antibacterial properties of P. regnellii justify its use in traditional medicine for the treatment of wounds, contaminated through bacteria infections.

Laboratory evaluation of methanolic extract of Atlantia monophylla (Family: Rutaceae) against immature stages of mosquitoes and non-target organisms

Methanolic extracts of the leaves of Atlantia monophylla (Rutaceae) were evaluated for mosquitocidal activity against immature stages of three mosquito species, Culex quinquefasciatus, Anopheles stephensi, and Aedes aegypti in the laboratory.Larvae of Cx. quinquefasciatus and pupae of An. stephensi were found more susceptible, with LC50 values of 0.14 mg/l and 0.05 mg/l, respectively. Insect growth regulating activity of this extract was more pronounced against Ae. aegypti, with EI50 value 0.002 mg/l. The extract was found safe to aquatic mosquito predators Gambusia affinis, Poecilia reticulata, and Diplonychus indicus, with the respective LC50 values of 23.4, 21.3, and 5.7 mg/l. The results indicate that the mosquitocidal effects of the extract of this plant were comparable to neem extract and certain synthetic chemical larvicides like fenthion, methoprene, etc.

In situ hybridization of hepatitis C virus RNA in liver cells of an experimentally infected rhesus macaque

The liver tissue of a rhesus macaque inoculated with hepatitis C virus (HCV) has been analyzed for the presence of HCV RNA using the technique of in situ hybridization, both at light and electron microscopy levels. The animal was inoculated by the intrasplenic route using a HCV infected autogenic hepatocyte transplant. The serum sample used to infect the hepatocyte cells was characterized by polymerase chain reaction technique and shown to be positive for HCV RNA, genotype 3 with 10(7) RNA copies/ml. In situ hybridization was performed using a complementary negative strand probe made with the specific primer. We were able to detect and localize viral RNA in altered membranes of the rough endoplasmic reticulum of infected liver cells, showing evidence of virus replication in vivo.

Respiratory viruses in the pediatric intensive care unit: prevalence and clinical aspects

A survey was conducted in two pediatric intensive care units in hospitals in Porto Alegre, Brazil, in order to monitor the main respiratory viruses present in bronchiolitis and/or pneumonia and their involvement in the severity of viral respiratory infections. Viral respiratory infection prevalence was 38.7%. In bronchiolitis, respiratory syncytial virus (RSV) was detected in 36% of the cases. In pneumonia, the prevalence rates were similar for adenovirus (10.3%) and RSV (7.7%). There was a difference among the viruses detected in terms of frequency of clinical findings indicating greater severity. Frequency of crackles in patients with RSV (47.3%) showed a borderline significance (p = 0.055, Fisher's exact test) as compared to those with adenovirus (87.5%). The overall case fatality rate in this study was 2.7%, and adenovirus showed a significantly higher case fatality rate (25%) than RSV (2.8%) (p = 0.005). Injected antibiotics were used in 49% of the children with RSV and 60% of those with adenovirus. Adenovirus was not detected in any of the 33 children submitted to oxygen therapy.

Isolation and identification of bovine tuberculosis in a Brazilian herd (São Paulo)

Mycobacterium was verified in animals from a Brazilian dairy herd, a total of 42 samples from 30 cows were submitted to culture and the isolated strains were analyzed by two polymerase chain reaction (PCR), the first specific for species belonging to the Mycobacterium complex (MTBC) and the other for differentiating M. tuberculosis from M. bovis. Twenty seven samples (64.3%) from 18 animals (60%) were positive for mycobacteria by culture, including samples from 15 retrofaryngeal lymphnodes (55.5%), 9 prescapular lymphnodes (33.3%), 2 lungs (7.4%), and 1 liver (3.7%). All isolated colonies were confirmed by PCR to contain MTBC organisms, and were identified as M. bovis by the same methodology.

Incidence of myiasis in Panama during the eradication of Cochliomyia hominivorax (Coquerel 1858, Diptera: Calliphoridae) (2002-2005)

We present the results of a study on myiasis in Panama during the first years of a Cochliomyia hominivorax eradication program (1998-2005), with the aim of investigating the behavior of the flies that produce myiasis in animals and human beings. The hosts that registered positive for myiasis were cattle (46.4%), dogs (15.3%), humans (14.7%), birds (12%), pigs (6%), horses (4%), and sheep (1%). Six fly species caused myiasis: Dermatobia hominis (58%), Phaenicia spp. (20%), Cochliomyia macellaria (19%), Chrysomya rufifacies (0.4%), and maggots of unidentified species belonging to the Sarcophagidae (3%) and Muscidae (0.3%). With the Dubois index, was no evidence that the absence of C. hominivorax allowed an increase in the cases of facultative myiasis.

Adenovirus, calicivirus and astrovirus detection in fecal samples of hospitalized children with acute gastroenteritis from Campo Grande, MS, Brazil

We analyzed fecal samples from hospitalized children up to three years of age with acute gastroenteritis at Campo Grande, Mato Grosso do Sul, Brazil, from May 2000-January 2004. Astrovirus and calicivirus were detected by Reverse Transcription-Polymerase Chain Reaction and adenovirus was detected using the Rotavirus and Adenovirus combined immunoenzyme assay. Astrovirus, adenovirus and calicivirus were detected at rates of 3.1%, 3.6% and 7.6%, respectively. These results re-emphasize the need for the establishment of regional vigilance systems to evaluate the impact of enteric viruses on viral gastroenteritis.

Evaluation of an enzyme-linked immunoelectrotransfer blot test for the confirmatory serodiagnosis of human toxocariasis

To improve the serodiagnosis of human toxocariasis, a sensitive and specific enzyme-linked immunoelectrotransfer blot (EITB-IgG) test was developed and evaluated using Toxocara canislarvae excretory-secretory antigens for detecting anti-Toxocara IgG antibodies. The EITB-IgG profile of toxocariasis was characterized by comparing 27 sera from patients with toxocariasis, 110 sera from healthy subjects and 186 sera from patients with other helminth diseases (ascariasis, ancylostomiasis, trichuriasis, enterobiasis, strongyloidiasis, hymenolepiasis, diphyllobothriasis, taeniasis, cysticercosis, hydatidosis and fascioliasis). Antigenic bands of 24, 28, 30, 35, 56, 117, 136 and 152 kDa were predominantly recognized in sera from all patients with toxocariasis. However, only bands of 24-35 kDa were highly specific for Toxocara infection (98.3%), whereas other antigenic bands observed displayed cross-reactivity. Additionally, when the results of the EITB-IgG test were compared to those of the ELISA-IgG test, a 100% concordance was observed for positive results in human toxocariasis cases. The concordance for negative results between the two tests for healthy subjects and patients with other helminth diseases were 96.3% and 53.7%, respectively, showing that the EITB-IgG test has a higher specificity than ELISA. In conclusion, the EITB-IgG test is a very useful tool to confirm the serological diagnosis of human toxocariasis.

Risk factors associated with human papillomavirus infection in two populations from Rio de Janeiro, Brazil

We investigated human papillomavirus (HPV) infection in two female populations from diverse socio-economic strata from the state of Rio de Janeiro and we also investigated the possible co-factors related to infection and the progression to cancer. In Group I, the reference group of this study, 10.7% of the patients presented HPV infection, as detected by generic PCR, while in Group II (low socio-demographic conditions) HPV was detected in 31.1% of the samples. HPV16 was the most prevalent virus type found in both Groups I and II (5.3% and 10%, respectively), followed by HPV 18 (1.3% and 4.7%, respectively). Although only a small sample was analysed, we detected differences among the groups regarding the rates of HPV infection, HPV types, age, ethnicity, familial income, schooling, marital status, parity, tobacco smoking and oral contraceptive use. For Group I, the Papanicolaou test was the most powerful independent factor associated with HPV status, followed by an age of under 30 years old, the number of sexual partners and black ethnicity. Our data are in agreement with the co-factors that are typically described for the developed world. For Group II, the Pap test was also the most relevant variable that was analysed, but the history of other sexually transmitted diseases and the use of alcohol were additional factors that were implicated in infection. These findings point out the need for the development of general and specific strategies for HPV screening of all Brazilian women.

Identification of clinical, epidemiological and laboratory risk factors for leprosy reactions during and after multidrug therapy

This cross-sectional retrospective study evaluated 440 leprosy patients; 57% (251/440) had leprosy reactions during and/or after multidrug therapy, 80.5% (202/251) of whom presented with multibacillary leprosy. At diagnosis, positive bacterial index (BI) [odds ratio (OR) = 6.39; 95% confidence interval (CI): 4.1-10.1)] or polymerase chain reaction (PCR) (OR = 9.15; 95% CI: 5.4-15.5) in skin smears, anti-phenolic glycolipid-1 (anti-PGL-1) ELISA (OR = 4.77; 95% CI: 2.9-7.9), leucocytosis (OR = 9.97; 95% CI: 3.9-25.7), thrombocytopenia (OR = 5.72; 95% CI: 2.3-14.0) and elevated lactate dehydrogenase (OR = 2.38; 95% CI: 1.4-4.0) were potential markers for the development of reactions during treatment. After treatment, positive BI (OR = 8.47; 95% CI: 4.7-15.3) and PCR (OR = 6.46; 95% CI: 3.4-12.3) in skin smears, anti-PGL-1 ELISA (OR = 2.25; 95% CI: 1.3-3.9), anaemia (OR = 2.36; 95% CI: 1.2-4.5), leucocytosis (OR = 4.14; 95% CI: 1.5-11.6) and thrombocytopenia (OR = 3.70; 95% CI: 1.3-2.2) were risk factors for the occurrence of reactions during the study period. The identification of groups with an increased risk for developing reactions will allow for the timely development of a treatment plan to prevent nerve damage and, therefore, the appearance of the disabling sequelae associated with the stigma of leprosy.

O banco do futuro: perspectivas e desafios

Neste artigo, apresentam-se as perspectivas e os desafios dos bancos no futuro. Com o referencial teórico sobre o tema, construiu-se um questionário para identificar o grau de importância atribuído a 20 diferentes questões. Na pesquisa de campo, abarcaram-se 93 bancários dos níveis gerencial e operacional dos principais bancos brasileiros e 9 professores universitários especialistas em mercado financeiro. Os dados coletados foram analisados estatisticamente usando-se o software Statistical Package for the Social Sciences, versão 13.0. Observando-se as principais conclusões da amostra, não há diferença estatística entre os funcionários dos bancos pesquisados, mas o mesmo não ocorreu entre bancários e professores. Algumas variáveis apresentaram maior relevância: combate às fraudes, business intelligence, bancarização e atendimento rápido. A partir da análise fatorial, identificaram-se seis fatores: sustentabilidade e papel do Brasil; mobilidade e segurança; regulação e novas tecnologias; globalização, inserção e privacidade; atendimento inteligente e bancarização. Pela análise discriminante, classificaram-se corretamente 79,3 e 66,7% dos funcionários do Bradesco e do Banco do Brasil, respectivamente, e 78,8% dos gerentes.