RESUMO
Using nonradioactive hybridization, the multilocal probes 33.15, F10 and (CAC)5 were shown to recognize multiple minisatellite regions in nuclear DNA of Leishmania, producing on Southern blots complex banding patterns typical of DNA fingerprints. We used the 33.15 probe to study 14 different strains belonging to 6 different species from both the sub-genus Viannia and the Leishmania mexicana complex of the sub-genus Leishmania. Distinct DNA fingerprints were obtained for each strain, permitting their identification. On the other hand, each strain showed little or no clonal variation. The information from the fingerprinting maps could be used for constructing phenograms and cladograms of the species and strains of Leishmania.
Assuntos
Impressões Digitais de DNA , DNA de Protozoário/genética , Leishmania/classificação , Leishmania/genética , Animais , Sondas de DNA , DNA Satélite/genética , Especificidade da EspécieRESUMO
To investigate the effect of lung volume on chest wall and lung mechanics in the rats, we measured the impedance (Z) under closed- and open-chest conditions at various positive end-expiratory pressures (0-0.9 kPa) by using a computer-controlled small-animal ventilator (T. F. Schuessler and J. H. T. Bates. IEEE Trans. Biomed. Eng. 42: 860-866, 1995) that we have developed for determining accurately the respiratory Z in small animals. The Z of total respiratory system and lungs was measured with small-volume oscillations between 0.25 and 9.125 Hz. The measured Z was fitted to a model that featured a constant-phase tissue compartment (with dissipation and elastance characterized by constants G and H, respectively) and a constant airway resistance (Z. Hantos, B. Daroczy, B. Suki, S. Nagy, and J. J. Fredberg. J. Appl. Physiol. 72: 168-178, 1992). We matched the lung volume between the closed- and open-chest conditions by using the quasi-static pressure-volume relationship of the lungs to calculate Z as a function of lung volume. Resistance decreased with lung volume and was not significantly different between total respiratory system and lungs. However, G and H of the respiratory system were significantly higher than those of the lungs. We conclude that chest wall in rats has a significant influence on tissue mechanics of the total respiratory system.
Assuntos
Pulmão/anatomia & histologia , Mecânica Respiratória/fisiologia , Tórax/fisiologia , Resistência das Vias Respiratórias/fisiologia , Algoritmos , Animais , Pulmão/fisiologia , Medidas de Volume Pulmonar , Masculino , Ratos , Ratos Sprague-DawleyRESUMO
The purpose of the present study was to establish how the dependence of respiratory mechanics on lung inflation changes during development. We studied seven groups of rats from 10 days to 3 mo of age at five levels of positive end-expiratory pressure (PEEP) from 0 to 7 hPa (1 hPa = 0.1 kPa approximately 1 cmH(2)O). At each PEEP level, we measured respiratory system resistance and elastance at both 0.9 and 4.8 Hz to partition the mechanical properties into its airway and tissue components. Elastance increased more rapidly with PEEP in the younger animals, which we interpret as reflecting a more pronounced strain stiffening of the younger parenchyma. However, the decrease in airway resistance with PEEP was more pronounced in the older animals. Morphometric analysis showed that mean tissue density decreased and total alveolar surface area increased with age. Our data suggest that the mechanical interdependence between airways and parenchyma is weaker in very young animals compared with mature animals. This may play a role in the hyperresponsiveness of immaturity.
Assuntos
Pulmão/crescimento & desenvolvimento , Pulmão/fisiologia , Respiração com Pressão Positiva , Mecânica Respiratória , Envelhecimento , Animais , Peso Corporal , Feminino , Pulmão/anatomia & histologia , Medidas de Volume Pulmonar , Masculino , Tamanho do Órgão , Alvéolos Pulmonares/crescimento & desenvolvimento , Alvéolos Pulmonares/fisiologia , Ratos , Ratos Sprague-DawleyRESUMO
Because of the wide utilization of rodents as animal models in respiratory research and the limited data on measurements of respiratory input impedance (Zrs) in small animals, we measured Zrs between 0.25 and 9.125 Hz at different levels (0-7 hPa) of positive end-expiratory pressure (PEEP) in mice, rats, guinea pigs, and rabbits using a computer-controlled small-animal ventilator (Schuessler TF and Bates JHT, IEEE Trans Biomed Eng 42: 860-866, 1995). Zrs was fitted with a model, including a Newtonian resistance (R) and inertance in series with a constant-phase tissue compartment characterized by tissue damping (Gti) and elastance (Hti) parameters. Inertance was negligible in all cases. R, Gti, and Hti were normalized to body weight, yielding normalized R, Gti, and Hti (NHti), respectively. Normalized R tended to decrease slightly with PEEP and increased with animal size. Normalized Gti had a minimal dependence on PEEP. NHti decreased with increasing PEEP, reaching a minimum at approximately 5 hPa in all species except mice. NHti was also higher in mice and rabbits compared with guinea pigs and rats at low PEEPs, which we conclude is probably due to a relatively smaller air space volume in mice and rabbits. Our data also suggest that smaller rodents have proportionately wider airways than do larger animals. We conclude that a detailed, comparative study of respiratory system mechanics shows some evidence of structural differences among the lungs of various species but that, in general, rodent lungs obey scaling laws similar to those described in other species.
Assuntos
Mecânica Respiratória/fisiologia , Roedores/fisiologia , Resistência das Vias Respiratórias/fisiologia , Animais , Cobaias , Camundongos , Camundongos Endogâmicos , Modelos Biológicos , Respiração com Pressão Positiva , Coelhos , Ratos , Ratos Sprague-Dawley , Respiração Artificial , Especificidade da EspécieRESUMO
DNA hybridisation was used to type 26 samples from lesions of human patients from the Rio Doce Valley (Minas Gerais, Brazil) clinically diagnosed as having cutaneous leishmaniasis, using kinetoplast DNA (kDNA) cloned mini-circle probes specific for the Leishmania mexicana and Leishmania braziliensis complexes. All samples were found to belong to the L. braziliensis complex. When biopsies were pressed directly onto touch blot membranes 38.5% of the samples were positive. The positivity and specificity obtained were both 100% when cultured blotted parasites were used. The results were confirmed by polymerase chain reaction (PCR) analysis using primers specific for the L. mexicana and L. braziliensis complexes.
Assuntos
DNA de Cinetoplasto/análise , DNA de Protozoário/análise , Leishmania braziliensis/genética , Leishmania mexicana/genética , Leishmaniose Cutânea/parasitologia , Animais , Biópsia , Brasil , DNA de Cinetoplasto/genética , DNA de Protozoário/genética , Doenças Endêmicas , Humanos , Leishmania braziliensis/química , Leishmania braziliensis/classificação , Leishmania mexicana/química , Leishmania mexicana/classificação , Leishmaniose Cutânea/diagnóstico , Leishmaniose Cutânea/epidemiologia , Hibridização de Ácido Nucleico , Reação em Cadeia da Polimerase , Sensibilidade e EspecificidadeRESUMO
This work reports the use of multidimensional HPLC by coupling a restricted access medium (RAM) bovine serum albumin (BSA) octadecyl column (100 x 4.6 mm I.D., 10 microm particle size and 120 A pore size) to an octadecyl Hypersil column (150 x 4.6 mm I.D., 5 microm particle size and 120 A pore size) to the analysis of amoxycillin in human plasma by direct injection. Ion pairing was necessary to extract amoxycillin with good recovery from the plasma proteins. To prepare the spiked samples, aliquots (60 microl) of the appropriated standard solutions were added to each culture tube containing an 180 microl of plasma and a solution of 0.30 mM tetrabuthylammonium phosphate (60 microl). They were vortexed for 15 s and then 290 microl were transferred to autosampler vials. Aliquots (250 microl) of the spiked plasma samples were injected to a column-switching HPLC system. An analysis time of 25 min with no time spent on sample preparation was achieved. The developed method showed good selectivity, sensitivity, accuracy and precision for direct analysis of this polar low wavelength ultraviolet absorption antibiotic using only 180 microl of human plasma. The validated method proved to be reliable and sensitive for the determination of amoxycillin in plasma samples of five healthy volunteers to whom test and reference formulations were administered as an oral dose (500 mg).
Assuntos
Amoxicilina/sangue , Cromatografia Líquida de Alta Pressão/métodos , Cromatografia Líquida de Alta Pressão/instrumentação , Humanos , Reprodutibilidade dos Testes , Espectrofotometria UltravioletaRESUMO
O trabalho teve como objetivo avaliar a influência de doses de cinzas em diferentes substratos no enraizamento de estacas de manjericão (Ocimum basilicum). O experimento foi conduzido em área do Núcleo de Capacitação e Pesquisa da Universidade Federal Rural da Amazônia, em Belém. Foi utilizado o delineamento experimental inteiramente casualizado, em esquema fatorial 2 x 5, constituído pelas combinações entre os fatores substratos (casca de arroz carbonizada e casca de castanha triturada) e acréscimo de cinzas (5 mL e 10 mL de cinza de casca de arroz, 5 mL e 10 mL de cinza de casca de castanha e sem cinza) com 4 repetições. Cada unidade experimental foi constituída por 10 estacas, num total de 400 estacas. Foram avaliadas as características percentual de enraizamento de estacas, número e comprimento de raízes. Não houve interações significativas entre os fatores avaliados (substrato e cinzas). Entre as características avaliadas o substrato casca de arroz carbonizada apresentou diferenças apenas no comprimento da raiz e no número de raízes, enquanto as dosagens de cinzas apresentaram diferenças apenas no percentual de enraizamento. O trabalho permitiu concluir que as dosagens de cinzas apresentaram efeito positivo sobre o enraizamento das estacas de manjericão, assim como o substrato a base de casca de arroz carbonizada propiciou melhores condições para o desenvolvimento das raízes. Assim, a casca de arroz é uma alternativa totalmente viável, pois além de apresentar características apropriadas para o enraizamento também possui baixo custo de aquisição, estando ao alcance de muitos produtores.
The study aimed to evaluate the influence of ash doses on different substrates on the stem rooting of basil (Ocimum basilicum). The experiment was conducted in the area of core training and research at the Universidade Federal Rural da Amazônia, in Belém, used a completely randomized design, factorial scheme 2 x 5 consisting of the combinations of the factors substrate (carbonized rice husk and bark brown ground) and increase ash (5 mL and 10 mL of rice husk ash 5 mL and 10 mL of husk ash and brown bark no gray) with four repetitions. Each experimental unit consisted of 10 cuttings, a total of 400 cuttings. Was evaluated the characteristics percentage of rooting, number and length of roots. There was no significant interaction between the factors evaluated (substrate and ash). Among the characteristics evaluated the carbonized rice showed only differences in root length and root number, while the levels of ash showed only differences in rooting percentage. The study revealed that the dosages of ash had a positive effect on rooting of cuttings of basil, as well as the substrate prepared with rice hulls provided the best conditions for root development. Thus, the rice husk is an alternative, as well as presenting features suitable for rooting also has a low cost of acquisition, being within reach of many producers.
Assuntos
Cinzas/métodos , Substratos para Tratamento Biológico/análise , Raízes de Plantas , Ocimum basilicum/crescimento & desenvolvimento , Casca de Planta/efeitos adversosRESUMO
Leishmania (Viannia) braziliensis isolates from two different geographic areas in Brazil were studied by DNA fingerprinting with the 33.15 multilocal probe and PCR with arbitrary primers (random amplification of polymorphic DNA-RAPD). The genetic distance of strains was measured by band sharing. The results showed that the strains isolated in Minas Gerais, southeastern Brazil, are very different from those isolated in Pará, northern Brazil. Strains from Minas Gerais constituted a relatively homogeneous group, presenting DNA fingerprint patterns with 76% of shared bands and RAPD profiles with 93% of shared bands. In contrast, strains from Pará showed higher genetic variability, with only 17 and 45% of shared bands in DNA fingerprint and RAPD patterns, respectively. This study suggests that genetic differences between L. braziliensis from both areas might have an epidemiological significance.
Assuntos
DNA de Protozoário/análise , Leishmania braziliensis/genética , Polimorfismo de Fragmento de Restrição , Animais , Sequência de Bases , Southern Blotting , Brasil , Impressões Digitais de DNA , Primers do DNA/química , DNA de Protozoário/química , Amplificação de Genes , Humanos , Leishmania braziliensis/classificação , Dados de Sequência Molecular , SoftwareRESUMO
Genetic relationships between Eimeria parasites of domestic fowl, including three isolates of E. acervulina, one of E. maxima, four of E. mitis, six of E. praecox, one of E. tenella, and two of uncertain identity, were analyzed by random amplified polymorphic DNA (RAPD) and the unweighted pair-group method with arithmetic mean, using 12 primers. Each primer amplified 4-34 DNA fragments/isolate. The similarity coefficients and phenograms were calculated from RAPD products with 230-2,000 bp in silver-stained polyacrylamide gels. Some primers generated RAPD markers which were species-specific for E. acervulina, E. mitis, and E. praecox. The phenograms revealed six clusters, each corresponding to an individual Eimeria species. The pBP and pBP2 isolates, of uncertain identity, clustered with the E. acervulina isolates (100% bootstrap). The intraspecific relationships showed certain degree of genetic isolation of the Eimeria populations, and it was associated with broiler house and with geographic separation.
Assuntos
Galinhas/parasitologia , Coccidiose/veterinária , DNA de Protozoário/análise , Eimeria/genética , Doenças das Aves Domésticas/parasitologia , Técnica de Amplificação ao Acaso de DNA Polimórfico , Animais , Coccidiose/parasitologia , Impressões Digitais de DNA , Primers do DNA/química , Eimeria/classificação , Eimeria/isolamento & purificação , Eletroforese em Gel de Poliacrilamida , Filogenia , Homologia de Sequência do Ácido Nucleico , Especificidade da EspécieRESUMO
O exame dos escritos de Freud permite a descoberta de uma invariavel em toda sua obra o inferir a si mesmo. Processo este que permite a avaliacao da constatacao do pouco interesse do psicanalista no trabalho institucional. O "habito de psicanalisar" e a manutencao do "mito da neutralidade", fora do "setting" analitico, sao fatos prejudiciais no engajamento do psicanalista na instituicao Repete assim, atraves da omissao, o mito edipico