Genetic stability of micropropagated plants of Crambe abyssinica Hochst using ISSR markers.

Crambe (Crambe abyssinica) is a non-edible annual herb, which was first cultivated to extract oil for industry, and now has great potential for biodiesel production. The objective of this investigation was to evaluate the genetic stability of micropropagated plants of the C. abyssinica Hochst cultivar 'FMS brilhante' using polymerase chain reaction techniques based on inter-simple sequence repeat (ISSR) molecular markers. The aim was to develop a protocol for the in vitro regeneration of these plants with low genetic variation as compared to the donor plant. For micropropagation, shoot tips from in vitro germinated seedlings were used as explants and were initially cultivated for 90 days on MS medium with 5.0 µM 6-benzylaminopurine (BAP), which at 90 days, led to the highest number of shoots per explant (NSE) (12.20 shoots) being detected. After 120 days, the interaction between BAP concentration and naphthalene acetic acid (NAA) was tested, and the highest NSE was observed following exposure to 0.0/0.5 µM BAP/NAA (11.40 shoots) and 1.0/0.0 µM BAP/NAA (11.00 shoots). The highest proportion of rooting phase were observed following exposure to 0.5 µM NAA (30%). The 13 ISSR primers used to analyze genetic stability produced 91 amplification products, of which only eight bands were polymorphic and 83 were monomorphic for all 10 regenerated crambe plants, compared to the donor plant explant. These results indicate that crambe shoot tips are a highly reliable explant that can be used to micropropagate genetically true-to-type plants or to maintain genetic stability, as verified using ISSR markers.

Influência de desfolhas artificiais para simular perdas na produção do feijoeiro (Phaseolus vulgaris L. cv. Xamego) / Influence of artificial defoliation to simulate losses in production of beans (Phaseolus vulgaris L. cv. Xamego)

A redução da área foliar é uma das causas da queda de produção em feijoeiro comum. O objetivo deste trabalho foi verificar a influência de diferentes níveis de desfolha em vários estádios de desenvolvimento sobre a produtividade, massa de 100 sementes e número de vagens por planta do feijoeiro da cultivar Xamego, em condições de campo no Sul do Espírito Santo. Utilizou-se o delineamento em blocos casualizados, sendo as parcelas dispostas no esquema fatorial 5 x 4: cinco épocas de desfolha (1ª folha trifoliolada, florescimento, formação das vagens, enchimento das vagem, e vagens secas) x quatro níveis de remoção das folhas (0, 33, 67 e 100%), com cinco repetições. Houve decréscimo linear da produtividade com os níveis de desfolha em todos os estádios de desenvolvimento, com exceção ao de vagens secas, sendo este decréscimo maior no estádio de formação das vagens. Pela análise de regressão atribui-se maior redução da produtividade com 100% de desfolha feita aos 44 dias após emergência. O componente de rendimento mais correlacionado com a produtividade foi o número de vagens por planta.

The reduction of foliar area can cause yield reduction in common bean crops. The objective of this work was to ascertain the effect of different defoliation levels realized in several developmental stages of common beans (Phaseolus vulgaris L. cv. Xamego) in regard to yield, weight of 100 seeds and number of pods per plant. A randomized complete block design was used considering a two-way 5 x 4 factorial arrangement: (i) five defoliation stages (first trifoliate leaf, flowering, pods formation, pods filling, and dry pods), and (ii) four defoliation levels (0, 33, 67 and 100%). Five replications were carried out. A linear decrease of yield was observed for all defoliation levels considering all development stages, except for dried pods. The highest decrease was detected for pods formation. At 100% of defoliation, a higher decrease of yield was observed when it was done at 44 days after emergence. Regression analysis gave a greater reduction in yield with 100% defoliation done at 44 days after emergence. The evaluation parameter that showed the best correlation with yield was the number of pods per plant.

Registro de ácaros em avestruz no estado do Rio de Janeiro / Report of mites in ostrich in Rio de Janeiro State

For the first time, it was identified a mite in a three-year-old male ostrich that had lost feathers and showed skin irritation. The animal belonged to a flock of 50 birds, located in Baixada Fluminense, in Rio de Janeiro State. After they were fixed in 70 percent alcohol, clarifyed in lactophenol and mounted in Canada balsam, the mites were identified in the laboratory. They belonged to the Pterolichidae family and Struthioperolichus sculpturatus species (Hirst). The ostrich was treated by ivermectin in dose of 200µg/kg of body weight, with recovery and disappearance of the clinical symptoms. The occurrence of mite infestation by the described species in ostrich should be monitored by professionals and farmers, as they cause fall of feathers and economic losses.

Systemic antibody response to diarrheagenic Escherichia coli and LPS O111, O157 and O55 in healthy Brazilian adults.

Enterohaemorrhagic Escherichia coli (EHEC) can cause a variety of human illnesses ranging from uncomplicated diarrhoea to haemorrhagic colitis and haemolytic uremic syndrome. The serotype O157:H7 has been associated with numerous outbreaks worldwide, but in Brazil the infection is rare. Brazilian adults present antibodies reactive with the principal virulence factors of enteropathogenic E. coli (EPEC) that have many genetic and antigenic similarities with EHEC. Lipopolysaccharides (LPS) are components of outer membranes and important virulence factors of Gram-negative bacteria. LPS O111 is present in EPEC and EHEC strains. LPS O157 is found only in EHEC strains, but it has some structural similarities with LPS O55 present in EPEC strains. This study investigates the levels of IgG and IgM seric antibodies reactive with EHEC O157:H7, EHEC O111:H-, EPEC O111:H- and the levels of anti-LPS O111, LPS O157 and LPS O55 antibodies in healthy adults living in São Paulo, Brazil. The antibody levels were determined by an enzyme-linked immunosorbent assay for 100 individual serum samples, and the presence of anti-bacterial and anti-LPS seric antibodies was confirmed. Positive correlations were found among the three kinds of antibodies. The concentrations of IgM anti-LPS were significantly higher than those of IgG, and surprisingly, the concentrations of anti-LPS O157 were high in view of the infrequent isolation of O157 bacteria in Brazil. Our results suggest that there is a cross-reacting immunity to EHEC in the Brazilian population, which may be a result of the immunity to EPEC antigens. Alternatively, Brazilians may be exposed to EHEC more frequently than has previously been thought.

IgA secretora total e específica no colostro e leite de mães da cidade do Natal-Rio Grande do Norte, Brasil / Total and specific IgA in colostrum and milk of mothers of Natal-Rio Grande do Norte, Brasil

PURPOSE: To determine the concentration of total secretory IgA and evaluate the repertoire of IgA antibodies to enteropathogenic Escherichia coli and Shigella flexneri antigens in colostrums and milk from mothers in Natal, RN. METHODS: The sample was constituted by 22 healthy clinically women whose babies were born at public hospital in Natal, RN. To determine total secretory IgA a radial immunedifusion tecnique (Mancini et al, 1965), was employed and to detect specific antibodies, immuneenzimatic assays, ELISA was used. RESULTS: The median values of total secretory IgA concentration presented individual variations with high levels in colostrums samples, decreasing during lactation, it was observed a p < 0.001 among the samples from the first day of lactation, to the thirtieth for total IgA concentration. All the donators present in colostrum and milk specific antibodies to Escherichia coli enteropathogenic (EPEC) and Shigella flexneri with titles higer in colostrum. There was parallel and directional pattern between total IgA and IgA anti-EPEC and Shegella flexneri, during period. CONCLUSION: The concentrations of total SIgA and specific antibodies to enteropathogenic Escherichia coli and Shigella flexneri in colostrums and milk in our study do not differ from others accomplished among populations with the same social and econimic features, stressing the importance of human milk as a protector agent against pathogens.

Treatment emergent affective switch: a controlled study.

OBJECTIVE: To study the clinical features of treatment emergent affective switch (TEAS) in comparison with spontaneous mania. METHODS: Twelve patients with TEAS within a 12-week period (average) of starting standard antidepressant medication were compared with 12 patients with spontaneous mania. RESULTS: Patients with TEAS were older, had longer duration of illness, more previous episodes, higher prevalence of subclinical hypothyroidism, and reported more previous episodes of mania associated with antidepressant use. TEAS was less severe, with a lower incidence of psychotic symptoms, lower Young Mania Rating Scale index score and rarely required hospitalization. The interval from intervention to response and remission was similar in both groups. CONCLUSION: TEAS was less severe, but had similar duration when compared with spontaneous mania. These results cannot directly answer the question of whether there is a causal relationship between antidepressant use and TEAS. While it is also possible that patients with longer duration of illness and higher cycle frequencies are more likely to experience episodes, it is difficult to attribute lesser severity of TEAS episodes to these clinical factors. Our observations are consistent with the suggestion that patients with longer duration of illness and previous history of TEAS may be at a greater risk of switching to mania during the use of antidepressants.

Anti-enteropathogenic Escherichia coli immunoglobulin Y isolated from eggs laid by immunised Leghorn chickens.

IgY, the egg yolk immunoglobulin, equivalent to the IgG from mammals, has been used in veterinary practice for passive immunisation against bacterial or viral infectious diseases. Enteropathogenic Escherichia coli (EPEC) is the main etiological agent of infantile diarrhoea in Brazil and other developing countries. Our aims were to isolate immunoglobulin IgY from egg yolk laid by EPEC -immunised Leghorn chickens and to study its reactivity to the antigens from this pathogen, including some virulence factors. Leghorn chickens were immunised with a bacterial suspension intramuscularly (three hens) or intravenously (three hens) or with PBS (two hens). Eggs were collected over a period of 17 weeks. IgY isolation procedures were carried out by salt precipitation (ammonium sulphate, in solid form) followed by centrifugations and dialysis. Final preparations were submitted to sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS - PAGE), enzyme-linked immunosorbent assay (ELISA) and immunoblotting. All immunised animals developed good levels of antibodies reactive to whole bacteria or lipopolysaccharide (LPS), in contrast to the control ones. Immunoblottings allowed the recognition of several antigenic fractions of bacterial antigens, some of which had a molecular weight compatible with bacterial virulence factors, confirming the efficacy of the immunisation and the adequacy of the method.

Lipid and acute-phase protein alterations in HIV-1 infected patients in the early stages of infection: correlation with CD4+ lymphocytes.

Lipid and acute-phase protein alterations have been described in various infection diseases, and they have been recorded during the early stages of HIV infection. Lipid and acute-phase protein profiles also have been correlated with cellular immunological abnormalities. To document these correlations during HIV infection, we studied 75 HIV-infected patients and 26 HIV-negative controls. Patients were classified according to the criteria proposed by the Walter Reed Army Institute: as WR-1 (CD4 lymphocytes, 1154 +/- 268/mm3), WR-2 (CD4, 793 +/- 348/mm3) and WR3/4 (CD4, 287+/-75 mm3). Triglycerides, total cholesterol and HDL-cholesterol concentrations were measured by enzymatic methods. Immunoglobulins (IgA and IgG) and acute-phase proteins (haptoglobin, alpha1-acid glycoprotein, C-reactive protein and transferrin) were determined by immunonephelometry. Haptoglobin levels were significantly increased in HIV-positive patients and correlated with the progression of HIV-infection (control

Effect of selected monoterpenes on methane oxidation, denitrification, and aerobic metabolism by bacteria in pure culture.

Selected monoterpenes inhibited methane oxidation by methanotrophs (Methylosinus trichosporium OB3b, Methylobacter luteus), denitrification by environmental isolates, and aerobic metabolism by several heterotrophic pure cultures. Inhibition occurred to various extents and was transient. Complete inhibition of methane oxidation by Methylosinus trichosporium OB3b with 1.1 mM (-)-alpha-pinene lasted for more than 2 days with a culture of optical density of 0.05 before activity resumed. Inhibition was greater under conditions under which particulate methane monooxygenase was expressed. No apparent consumption or conversion of monoterpenes by methanotrophs was detected by gas chromatography, and the reason that transient inhibition occurs is not clear. Aerobic metabolism by several heterotrophs was much less sensitive than methanotrophy was; Escherichia coli (optical density, 0.01), for example, was not affected by up to 7.3 mM (-)-alpha-pinene. The degree of inhibition was monoterpene and species dependent. Denitrification by isolates from a polluted sediment was not inhibited by 3.7 mM (-)-alpha-pinene, gamma-terpinene, or beta-myrcene, whereas 50 to 100% inhibition was observed for isolates from a temperate swamp soil. The inhibitory effect of monoterpenes on methane oxidation was greatest with unsaturated, cyclic hydrocarbon forms [e.g., (-)-alpha-pinene, (S)-(-)-limonene, (R)-(+)-limonene, and gamma-terpinene]. Lower levels of inhibition occurred with oxide and alcohol derivatives [(R)-(+)-limonene oxide, alpha-pinene oxide, linalool, alpha-terpineol] and a noncyclic hydrocarbon (beta-myrcene). Isomers of pinene inhibited activity to different extents. Given their natural sources, monoterpenes may be significant factors affecting bacterial activities in nature.

Methane metabolism in a temperate swamp.

Comparisons between in situ CH(4) concentration and potential factors controlling its net production were made in a temperate swamp. Seasonal measurements of water table level and depth profiles of pH, dissolved CH(4), CO(2), O(2), SO(4), NO(3), formate, acetate, propionate, and butyrate were made at two adjacent sites 1.5 to 2 m apart. Dissolved CH(4) was inversely correlated to O(2) and, in general, to NO(3) and SO(4), potential inhibitors of methanogenesis. At low water table levels (August 1992), maximal CH(4) (2 to 4 muM) occurred below 30 cm, whereas at high water table levels (October 1992) or under flooded conditions (May 1993), CH(4) maxima (4 to 55 muM) occurred in the top 10 to 20 cm. Higher CH(4) concentrations were likely supported by inputs of fresh organic matter from decaying leaf litter, as suggested by high acetate and propionate concentrations (25 to 100 muM) in one of the sites in fall and spring. Measurements of potential CH(4) production (and consumption) showed that the highest rates generally occurred in the top 10 cm of soil. Soil slurry incubations confirmed the importance of organic matter to CH(4) production but also showed that competition for substrates by nonmethanogenic microorganisms could greatly attenuate its effect.

Ca2+ binding to occluded sites in the CrATP-ATPase complex of sarcoplasmic reticulum: evidence for two independent high-affinity sites.

Occluded Ca2+ sites in the CrATP-ATPase complex are studied by first forming the complex in the presence of EGTA so that the sites can be occluded while vacant. 45Ca2+ binding to the occluded sites is then studied under equilibrium conditions. Binding curves are produced for two independent Ca2+ sites with Kd(1) = 0.2 microM and Kd(2) = 1.6 microM. When both sites are saturated, only the Ca2+ bound to the lower affinity site can exchange with free Ca2+. On addition of EGTA (15 vs 0.5 mM Ca2+) all bound Ca2+ dissociates, the net dissociation rate of one-half of the Ca2+ being approximately 10-fold greater than that of the other one-half (at 37 degrees C). When Ca2+ is bound only to the higher affinity site, this Ca2+ will exchange slowly if the concentration of free Ca2+ is below the saturation level of the lower affinity site. An ionophore dependency of the rates of binding and dissociation indicates that the access to the sites is through the interior of the vesicle. Solubilization in C12E9 releases the Ca2+ in the higher affinity site. Our observations are consistent with a model of the ATPase where the lower affinity of two transport sites is associated with the interior position (closest to the lumen) in a transmembrane channel. It is further evident that when in the occluded state, the higher affinity site is available without Ca2+ first being bound to the lower affinity site, eliminating cooperativity from the binding mechanism. In turn, this implies a connection between the integrity of the high-affinity binding site and the linking of sections of the catalytic site by CrATP.

Association between parental and student smoking behaviour in a Brazilian medical school.

In a 20% random sample of University of Brasilia medical students no association was found between student and parental smoking behaviour. A low prevalence of habitual smoking (14.0%) and a low mean daily cigarette consumption (8.9 cigarettes per smoker, per day) characterized this population.