Cytogenetic and identification of the nucleolus organizer region in Heliconia bihai (L.) L.

The genus Heliconia is not much studied and the number of existing species in this genus is still uncertain. It is known that this number relies between 150 to 250 species. In Brazil, about 40 species are native and known by many different names. The objective of this paper was to characterize morphometrically and to identify the NOR (active nucleolus organizer regions) by Ag-NOR banding of chromosomes of Heliconia bihai (L) L. Root meristems were submitted to blocking treatment in an amiprofos-methyl (APM) solution, fixed in methanol-acetic acid solution for 24 hours, at least. The meristems were washed in distilled water and submitted to enzymatic digestion with pectinase enzyme. The slides were prepared by dissociation of the root meristem, dried in the air and also on hot plate at 50°C. Subsequently, some slides were submitted to 5% Giemsa stain for karyotype construction and to a solution of silver nitrate (AgNO3) 50% for Ag-NOR banding. The species H. bihai has 2n = 22 chromosomes, 4 pairs of submetacentric chromosomes and 7 pairs of metacentric chromosomes, and graded medium to short (3.96 to 0.67 μM), with the presence of active NOR in pairs 1 and 2 and interphase cells with 2 nucleoli. These are the features of a diploid species.

Identification of 'Ubá' mango tree zygotic and nucellar seedlings using ISSR markers

Polyembryonic seeds are characterized by the development of over one embryo in the same seed, which can be zygotic and nucellar. The objective of this work was to identify the genetic origin, whether zygotic or nucellar, of seedlings of polyembryonic seeds of 'Ubá' mango tree using ISSR markers, and relating them with the vigor of the seedlings. Thus, mangos were harvested in Visconde do Rio Branco (accession 102) and Ubá (accessions 112, 138, 152 and 159), whose seeds were germinated in plastic trays filled with washed sand. Fifty days after sowing, seedlings from five seeds of each one of the accessions 102, 112, 138, 159 and from 10 seeds of the accession 152, were analyzed. These sseedlings were characterized and evaluated for plant height, stem circumference and mass of fresh aerial part and the most vigorous seedling was the one displaying at least two of these traits higher than the other seedlings from seed. Leaves were collected for genomic DNA extraction, which was amplified using seven ISSR primers previously selected based on the amplification profile and considering the number and resolution of fragments. Zygotic seedlings were found in 18 seeds, which were the most vigorous in six seeds. The results evidenced the existence of genetic variability in orchards using seedlings grown from seeds, because the farmer usually uses the most vigorous ones, assuming that this is of nucellar origin. These results also indicate that the most vigorous seedling are not always nucellar, inasmuch as of 20% of the total seeds evaluated, the zygotic seedling was the most vigorous.

Isolation and serological identification of enteropathogenic Escherichia coli in pasteurized milk in Brazil

OBJECTIVE: To evaluate the microbiological quality of pasteurized milk commercialized in Rio de Janeiro, Brazil, and determine serologically enteropathogenic Escherichia coli (EPEC) strains in E. coli isolates obtained from milk samples. METHODS: Ninety samples of pasteurized milk -- types B and C -- of three different commercial brands, purchased in supermarkets and bakeries in Rio de Janeiro, were examined. The amount of total and fecal coliform bacteria was estimated using the Most Probable Number technique. Mesophilic, psychrotrophic, and thermoduric microorganism counts were determined by the Standard Plate Count technique. Isolation and identification of E. coli were carried out using conventional physiological tests. Commercial antisera were used for serological characterization of EPEC. RESULTS: The three milk brands analyzed revealed bacterial counts above the regulated values of the Brazilian government. It was found that among 208 strains of E. coli isolated, 46 (22.1%) were serologically classified as EPEC. The most common EPEC serogroup was O55 (15.2%). CONCLUSIONS: Though recent studies on virulence factors indicate that not all strains serologically classified as EPEC are able to attaching/effacing lesion, it is believed that the isolation of EPEC serogroups from pasteurized milk represent a potential risk for children, as well as an indicative of the presence of other enteropathogens.

Seqüência de drogas consumidas por usuários de crack e fatores interferentes

OBJETIVO: Identificar, entre usuários de crack, uma progressão no uso de drogas e seus fatores interferentes. MÉTODOS: Utilizou-se metodologia qualitativa para uma investigação mais profunda, considerando o ponto de vista que o entrevistado tem do fenômeno. Foram aplicados entrevistas de longa duração e questionários semi-estruturados. Foi delineada uma amostra intencional, e uma amostragem com critérios foi conseguida. Para atingir a saturação teórica, foram entrevistados 31 usuários ou ex-usuários de crack. RESULTADOS: Foram detectadas duas fases distintas de uso de drogas. A primeira, com drogas lícitas, sendo o cigarro e o álcool as mais citadas pela amostra. Parentes e amigos dos entrevistados foram os incentivadores do consumo, e o motivo alegado para o uso dessas substâncias foi a necessidade de autoconfiança. A idade precoce do consumo e o uso pesado de uma ou ambas as drogas foram determinantes para o início de uma escalada de drogas ilícitas. A maconha foi a primeira droga dessa segunda fase. Uma postura mais ativa na busca da droga como fonte de prazer passou a ser o motivo do consumo. CONCLUSÕES: O estudo revela que a identificação de uma seqüência de drogas parece estar mais associada a fatores externos (pressões de grupo, influência do tráfico etc.) do que à preferência do usuário. Foram identificadas duas progressões diferentes: entre os mais jovens (=30 anos), cuja a escalada começou com o cigarro e/ou álcool e passou pela maconha e cocaína aspirada até o uso de crack; e os mais velhos (>30 anos), que iniciaram o uso de drogas pelo cigarro e/ou álcool, seguido de maconha, medicamentos endovenosos, cocaína aspirada, cocaína endovenosa e, por fim, crack.

Caracterização da cultura de crack na cidade de São Paulo: padrão de uso controlado

OBJETIVO: Caracterizar a situação do uso de crack na cidade de São Paulo, assim como o perfil sociodemográfico de seu usuário. PROCEDIMENTOS METODOLÓGICOS: Estudo qualitativo etnográfico com amostra intencional de usuários (n=45) e ex-usuários de crack (n=17). Os participantes foram recrutados pela técnica de amostragem em cadeias e responderam a uma entrevista semi-estruturada, direcionada por questionário, durante os anos de 2004 e 2005. O conjunto de cada questão e suas respectivas respostas originou relatórios específicos que foram interpretados individualmente. ANÁLISE DOS RESULTADOS: O perfil predominante do usuário de crack foi ser homem, jovem, solteiro, de baixa classe socioeconômica, baixo nível de escolaridade e sem vínculos empregatícios formais. O padrão de uso mais freqüentemente citado foi o compulsivo, caracterizado pelo uso múltiplo de drogas e desenvolvimento de atividades ilícitas em troca de crack ou dinheiro. Entretanto, identificou-se o uso controlado que consiste no uso não-diário de crack, mediado por fatores individuais, desenvolvidos intuitivamente pelo usuário e semelhantes, em natureza, às estratégias adotadas por ex-usuários para o alcance do estado de abstinência. CONCLUSÕES: A cultura do uso de crack tem sofrido mudanças quanto ao padrão de uso. Embora a maioria dos usuários o faça de forma compulsiva, observou-se a existência do uso controlado, que merece maior detalhamento, principalmente quanto às estratégias adotadas para seu alcance.

HIV/AIDS risk among female sex workers who use crack in Southern Brazil

OBJECTIVE: To understand the social context of female sex workers who use crack and its impact on HIV/AIDS risk behaviors. METHODODOLOGICAL PROCEDURES: Qualitative study carried out in Foz do Iguaçu, Southern Brazil, in 2003. Twenty-six in-depth interviews and two focus groups were carried out with female commercial sex workers who frequently use crack. In-depth interviews with health providers, community leaders and public policy managers, as well as field observations were also conducted. Transcript data was entered into Atlas.ti software and grounded theory methodology was used to analyze the data and develop a conceptual model as a result of this study. ANALYSIS OF RESULTS: Female sex workers who use crack had low self-perceived HIV risk in spite of being engaged in risky behaviors (e.g. unprotected sex with multiple partners). Physical and sexual violence among clients, occasional and stable partners was widespread jeopardizing negotiation and consistent condom use. According to health providers, community leaders and public policy managers, several female sex workers who use crack are homeless or live in slums, and rarely have access to health services, voluntary counseling and testing, social support, pre-natal and reproductive care. CONCLUSIONS: Female sex workers who use crack experience a plethora of health and social problems, which apparently affect their risks for HIV infection. Low-threshold, user-friendly and gender-tailored interventions should be implemented, in order to increase the access to health and social-support services among this population. Those initiatives might also increase their access to reproductive health in general, and to preventive strategies focusing on HIV/AIDS and other sexually transmitted infections.

Evolução do consumo de crack em coorte com histórico de tratamento

OBJETIVO: Analisar a evolução do consumo entre usuários de crack com histórico de tratamento. MÉTODOS: Uma coorte de, originalmente, 131 dependentes de crack admitidos em uma enfermaria de desintoxicação em São Paulo, SP, entre 1992 e 1994, foi re-entrevistada em três ocasiões: 1995-1996, 1998-1999 e 2005-2006. As variáveis averiguadas foram: dados demográficos, comportamento sexual de risco, padrões de consumo de crack e outras substâncias, prisões, desaparecimentos e óbitos. Na análise estatística empregou-se o teste de qui-quadrado, a regressão logística multinomial e regressão de Cox. RESULTADOS: Dos pacientes avaliados, 43 estavam abstinentes do crack (12 meses ou mais), 22 eram usuários, 13 estavam presos, dois desaparecidos e 27 estavam mortos. Foram identificados três grupos com trajetórias distintas de consumo pós-alta. Comportamento seguro com uso de preservativo foi identificado como fator relacionado ao grupo de abstinentes estáveis (p = 0,001). Teste HIV positivo na internação (p = 0,046); consumo de cocaína aspirada no último ano (p = 0,001) e tempo de uso de cocaína aspirada na vida (mais de 132 meses) (p = 0,000) foram fatores relacionados a uso de longo termo. Uso pregresso de cocaína endovenosa aumentou em 2,5 vezes as chances de óbito em 12 anos (p = 0,031) (IC95%: 1,08; 5,79). CONCLUSÕES: A recorrência e persistência do consumo nos anos pós-alta de tratamento refletem novas modalidades de uso do crack. Por outro lado, padrões de abstinência estável apontam a viabilidade dos processos de recuperação relativos ao uso de crack.

Fissura por crack: comportamentos e estratégias de controle de usuários e ex-usuários

OBJETIVO: Compreender a fissura do usuário de crack, bem como descrever os comportamentos desenvolvidos sob fissura e estratégias utilizadas para seu controle. PROCEDIMENTOS METODOLÓGICOS: Estudo qualitativo com amostra intencional por critérios de 40 usuários e ex-usuários de crack em São Paulo, SP, nos anos de 2007 e 2008. Os entrevistados foram recrutados pela técnica da bola de neve e submetidos à entrevista semi-estruturada em profundidade, até a saturação teórica. Após transcrição literal, seguiu-se a análise do conteúdo das entrevistas para elaboração de inferências e hipóteses alicerçadas nessas narrativas. ANÁLISE DOS RESULTADOS: Os entrevistados estavam igualmente distribuídos quanto ao sexo, possuíam idade entre 18 e 50 anos, abrangeram todos os níveis de escolaridade e a maior parte possuía poucos recursos financeiros. Além da fissura sentida na abstinência de crack e da fissura induzida por pistas ambientais e emocionais, constatou-se um tipo de fissura que faz parte do próprio efeito do crack. Esta última apareceu como forte fator mantenedor dos binges de consumo, que foram os maiores responsáveis pelo rebaixamento de valores do usuário, sujeitando-o a práticas arriscadas para a obtenção da droga. Os métodos mais citados para a obtenção de crack ou dinheiro para comprá-lo foram: prostituição, manipulação de pessoas, endividamento, troca de pertences por crack e roubo. Foram relatadas estratégias para o alívio da fissura e táticas farmacológicas e comportamentais para evitar o seu desenvolvimento, como: comer, ter relação sexual, jogar futebol, trabalhar, evitar o contexto social de uso de crack e usar drogas que causam sonolência. CONCLUSÕES: Os binges de consumo de crack são causados pela fissura induzida durante o uso da droga. As medidas criadas pelo próprio usuário para lidar com a sua fissura melhoram sua relação com o crack e podem ser ferramenta importante para o aprimoramento do tratamento.

Isolation of human fungi from soil and identification of two endemic areas of Cryptococcus neoformans and Coccidioides immitis

The present study was carried out in two different areas of Province of Cordoba, Argentina, where there was a suspicious of endemic mycosis. The previous data were the presence of a clinical case of pulmonary cryptococcosis in one area (Alta Gracia) and the previous findings of a high incidence of coccidioidin and cryptococcin reactors in the population of the second one (Villa Dolores). In both areas soil samples for fungi were studied and Cryptococcus neoformans was found in 2/25 samples from Alta Gracia. In Villa Dolores Coccidioides immitis was isolated in 2/40 samples, and C. neoformans in 1/40 samples. Delayed hypersensitivity test with cryptococcin was determined in the population from Alta Gracia and it was found to be 5.3%. Positive cutaneous tests with coccidioidin (33.8%) and cryptococcin (31.9%) in Villa Dolores were obtained. With these findings two endemic areas of systemic mycoses in Cordoba, Argentina were delimited.

Simultaneous identification of Trypanosoma cruzi surface and internal antigens reactive to different immunoglobulin classes (radio-immunoblotting)

A radioactive Western-blotting technique was developed by which the reactivity of Immunoglobulins (Igs) from different classes to both membrane radiolabelled and internal parasite antigens is simultaneously identified. The method includes radioiodination of parasites, polypeptide fractionation by SDS-PAGE, Western-blot transfer and autoradiography of the immunoblots developed with anti-Igs conjugates labelled with enzymes. The analysis is then performed by the comparison of common bands on the autoradiograms and the respective substrate stained nitrocellulose blots. This technique was used to analyse T. cruzi trypomastigote surface labelled antigens reactive to IgM, IgA and IgG specific antibodies. A different pattern of reactivity with acute Chagas' disease patients sera was thus obtained.

Schistosoma mansoni: identification of a 46KDa antigen of the schistosomular surface by monoclonal antibody

An IgG2a subclass monoclonal antibody, C6G9, was obtained by immunization of BALB/c mice with Schistosoma mansoni egg antigens. With this monoclonal antibody, it was possible to identify a schistosomular antigen with a molecular weight of 46 kilodaltons (KDa), and its expression being evaluated by means of indirect immunofluorescence. The antigen persisted in the integument of the developing schistosomulum, for at least 96 hours post-transformation. The monoclonal antibody also reacted with the cercaria surface, but not with that of adult worm. The C6G9 was also able to mediate significant levels of cytotoxicity in the presence of complement for newly transformed schistosomula.

Identification of factors and groups at risk of infection with Schistosoma mansoni: a strategy for the implementation of control measures?

A fourteen year schistosomiasis control program in Peri-Peri (Capim Branco, MG) reduced prevalence from 43.5 to 4.4%; incidence from 19.0 to 2.9%, the geometric mean of the number of eggs from 281 to 87 and the level of the hepatoesplenic form cases from 5.9 to 0.0%. In 1991, three years after the interruption of the program, the prevalence had risen to 19.6%. The district consists of Barbosa (a rural area) and Peri-Peri itself (an urban area). In 1991, the prevalence in the two areas was 28.4% and 16.0% respectively. A multivariate analysis of risk factors for schistosomiasis indicated the domestic agricultural activity with population attributive risk (PAR) of 29.82%, the distance (< 10 m) from home to water source (PAR = 25.93%) and weekly fishing (PAR = 17.21%) as being responsible for infections in the rural area. The recommended control measures for this area are non-manual irrigation and removal of homes to more than ten meters from irrigation ditches. In the urban area, it was observed that swimming at weekly intervals (PAR = 20.71%), daily domestic agricultural activity (PAR = 4.07%) and the absence of drinking water in the home (PAR=4.29%) were responsible for infections. Thus, in the urban area the recommended control measures are the substitution of manual irrigation with an irrigation method that avoids contact with water, the creation of leisure options of the population and the provision of a domestic water supply. The authors call attention to the need for the efficacy of multivariate analysis of risk factors to be evaluated for schistosomiasis prior to its large scale use as a indicator of the control measures to be implemented.

Human cutaneous leishmaniasis caused by Leishmania (Viannia) braziliensis in Santiago del Estero, Argentina: identification of parasites by monoclonal antibodies and isoenzymes

Diagnostic and parasite characterization and identification studies were carried out in human patients with cutaneous leishmaniasis lesions in Santiago del Estero, Northern Province of Argentina. Diagnostic procedures were biopsies of lesions for smears and inoculations in hamster, needle aspirations of material from ulcers for "in vitro" cultures. Immunodiagnostic techniques applied were IFAT-IgG and Montenegro skin test. Primary isolation of eight stocks of leishmanial parasites was achieved from patients with active lesions. All stocks were biologically characterized by their behaviour in hamster, measurements of amastigote and promastigotes and growth "in vitro". Eight stocks were characterized and identified at species level by their reactivity to a cross-panel of sub-genus and specie-specific Monoclonal Antibodies through an Indirect Immunofluorescence technique and a Dot-ELISA. We conclude from the serodeme analysis of Argentina stocks that: stocks MHOM/AR/92/SE-1; SE-2; SE-4; SE-8; SE-8-I; SE-30; SE-34 and SE-36 are Leishmania (Viannia) braziliensis. Three Leishmania stocks (SE-1; SE-2 and SE-30) did not react with one highly specie-specific Monoclonal Antibody (Clone: B-18, Leishmania (Viannia) braziliensis marker) disclosing two serodeme group patterns. Five out of eight soluble extracts of leishmanial promastigotes were electrophoresed on thin-layer starch gels and examined for the enzyme MPI, Mannose Phosphate Isomerase; MDH, Malate Dehydrogenase; 6PGD, 6 Phosphogluconate Dehydrogenase; NH, Nucleoside Hydrolase, 2-deoxyinosinc as substrate; SOD, Superoxide Dismutase; GPI, Glucose Phosphate Isomerase and ES, Esterase. From the isoenzyme studies we concluded that stocks: MHOM/AR/92/SE-1; SE-2; SE-4; SE-8 and SE-8-I are isoenzymatically Leishmania (Viannia) braziliensis. We need to analyze more enzymes before assigning them to a braziliensis zymodeme.

EVALUATION OF A RAPID SCREENING ASSAY FOR BACTERIAL IDENTIFICATION (DOT-ELISA) IN FECAL SAMPLES FROM CHILDREN

With the objective of standardizing a Dot Enzyme-Linked Immunosorbent Assay (Dot-ELISA) to detect antigens of fecal bacterial enteropathogens, 250 children, aged under 36 months and of both sexes, were studied; of which 162 had acute gastroenteritis. The efficacy of a rapid screening assay for bacterial enteropathogens (enteropathogenic Escherichia coli "EPEC", enteroinvasive Escherichia coli "EIEC", Salmonella spp. and Shigella spp.) was evaluated. The fecal samples were also submitted to a traditional method of stool culture for comparison. The concordance index between the two techniques, calculated using the Kappa (k) index for the above mentioned bacterial strains was 0.8859, 0.9055, 0.7932 and 0.7829 respectively. These values express an almost perfect degree of concordance for the first two and substantial concordance for the latter two, thus enabling this technique to be applied in the early diagnosis of diarrhea in infants. With a view to increasing the sensitivity and specificity of this immunological test, a study was made of the antigenic preparations obtained from two types of treatment: 1) deproteinization by heating; 2) precipitation and concentration of the lipopolysaccharide antigen (LPS) using an ethanol-acetone solution, which was then heated in the presence of sodium EDTA

Detection and identification of dengue virus isolates from Brazil by a simplified reverse transcription - polymerase chain reaction (RT-PCR) method

We show here a simplified RT-PCR for identification of dengue virus types 1 and 2. Five dengue virus strains, isolated from Brazilian patients, and yellow fever vaccine 17DD as a negative control, were used in this study. C6/36 cells were infected and supernatants were collected after 7 days. The RT-PCR, done in a single reaction vessel, was carried out following a 1/10 dilution of virus in distilled water or in a detergent mixture containing Nonidet P40. The 50 µl assay reaction mixture included 50 pmol of specific primers amplifying a 482 base pair sequence for dengue type 1 and 210 base pair sequence for dengue type 2. In other assays, we used dengue virus consensus primers having maximum sequence similarity to the four serotypes, amplifying a 511 base pair sequence. The reaction mixture also contained 0.1 mM of the four deoxynucleoside triphosphates, 7.5 U of reverse transcriptase, 1U of thermostable Taq DNA polymerase. The mixture was incubated for 5 minutes at 37ºC for reverse transcription followed by 30 cycles of two-step PCR amplification (92ºC for 60 seconds, 53ºC for 60 seconds) with slow temperature increment. The PCR products were subjected to 1.7% agarose gel electrophoresis and visualized by UV light after staining with ethidium bromide solution. Low virus titer around 10 3, 6 TCID50/ml was detected by RT-PCR for dengue type 1. Specific DNA amplification was observed with all the Brazilian dengue strains by using dengue virus consensus primers. As compared to other RT-PCRs, this assay is less laborious, done in a shorter time, and has reduced risk of contamination