Selecting sugarcane genotypes by the selection index reveals high gain for technological quality traits.

Sugarcane (Saccharum sp) is one of the most promising crops and researchers have sought for renewable alternative energy sources to reduce CO2 emission. The study of strategies, which allow breeders in the selection of superior genotypes for many traits simultaneously, is important. Therefore, the objectives of this study were: i) to apply path analysis to better understand the relationship between the lignocellulosic traits and technological quality traits with total recoverable sugars (TRS) and ii) to use several multivariate selection indexes to predict the genetic gain and to select superior genotypes in the sugarcane breeding. A total of 40 sugarcane genotypes were evaluated in an experimental design using incomplete blocks with two replicates. The follow traits were evaluated: dry matter (DM), total soluble solids (BRIX), apparent sucrose content in the juice (POL), apparent sucrose content in sugarcane (POLS), fiber content (FIB), purity (PUR), TRS, lignin content (LC), cellulose content (CC), hemicellulose content (HC), and ash content (AC). These traits were analyzed by analysis of variance, phenotypic correlation network, path analysis, and selection index. The highest direct effect on TRS was obtained by POLS (0.337), POL (0.299), BRIX (0.227), and FIB (-0.146). The estimates of phenotypic correlation between these characters and TRS were in the same direction, which demonstrated a cause-and-effect relationship. The highest indirect effect was of POL via POLS (0.331) followed by POLS via POL (0.294). BRIX presented high indirect effects via POLS (0.266) and via POL (0.246). On the other hand, FIB presented negative indirect effects via POLS (-0.169) and POL (-0.103). In conclusion, path analysis and index selection are useful strategies to help breeders in the selection of superior genotypes in sugarcane.

Development of a method for extraction and assay of human erythrocyte acetylcholinesterase and pesticide inhibition.

A method of extracting membranes from red blood cells (RBCs) is described, which were in turn used to assay acetylcholinesterase (AChE) activity. The evidence for the enzyme activity was established by selective inhibition using 1,5-bis(4-allyldimethylammoniumphenyl) pentan-3-one dibromide, tetraisopropyl pyrophosphoramide and neostigmine. Blood samples were exposed to three organophosphorus (dichlorvos, chlorpyrifos and diazinon) and two carbamate (carbaryl and carbofuran) pesticides. Afterwards AChE activities in RBC membranes were determined. The concentrations capable to inhibit the enzyme activity by 50% (IC50) for the pesticides were 10.66 µM (dichlorvos), 21.42 µM (chlorpyrifos), 109.98 µM (carbaryl) and 5.44 µM (carbofuran). The results related to 20% enzyme inhibition (level used in the estimation of threshold limits for anticholinesterase compounds) were below those acceptable daily intake values enacted by relevant national and international regulations. These results suggest that the proposed AChE extraction from RBC and assay could be a suitable method for monitoring occupational exposure to pesticides.

Levels of oxidative stress markers: correlation with hepatic function and worm burden patients with schistosomiasis.

Schistosomiasis is caused by Schistosoma mansoni and is a public health problem in Brazil. The typical granulomatous lesion is associated with the increase in the oxidative damage by generation of free radicals. The aim of this work was to correlate some oxidative stress markers with the worm burden on carriers of schistosomiasis (n = 30) in the acute phase in comparison to healthy subjects (n = 30). The pro-oxidant parameter used was the colorimetric quantification of reactive substances to thiobarbituric acid, while the antioxidant markers used were blood content of reduced glutathione and determination of the activity of catalase. The worm burden was assessed by Kato-Katz method. The results pointed out that initially there was no difference in the catalase activity. However, there was a positive correlation between the increase in parasitic load and intensity of lipid peroxidation, and decrease in the content of reduced glutathione. Additionally, only the aspartate aminotransferase levels presented to be high, while there was a decrease in bilirubin level. Therefore, a possible association between the establishment of the oxidative stress in tissue and the parasitic load of Schistosoma mansoni is suggested.

The demands of family caregivers of elderly individuals with dementia / Demanda do cuidados familiar com idoso demenciado / Demandas del cuidador familiar del anciano con demencia

The purpose of this cross-sectional observational study was to identify characteristics of caregivers of elderly people with dementia, types of care demands and to relate demands to the stage of dementia. The study was carried out in 2004 with 104 older adults and 90 caregivers in Ribeirão Preto, state of São Paulo. The OARS instrument was utilized and a questionnaire answered by caregivers. Among older adults, 66.3 percent were female, aged 75.5 years in average and 86.5 percent had caregivers. Eighty percent of the caregivers were women family members, aged 52.3 years in average. They spent 15.10 hours/day with care, without help. An important relationship was observed between caregiver's burden, physical and emotional effort and stage of dementia. Emotional overburden was higher at dementia early and late stages, this difference was statistically non-significant. Results reveal the urgent need to plan formal and informal support strategies to caregivers of Brazilian elderly people with dementia.

As finalidades deste estudo foram identificar as características dos cuidadores de idosos demenciados, os tipos de demandas de cuidados e relacionar a demanda com o estágio da demência. O estudo seccional e observacional foi conduzido em 2004, com 104 idosos e 90 cuidadores, em Ribeirão Preto/SP, por meio do instrumento OARS e de um questionário para o cuidador. Dos 104 idosos, 66.3 por cento eram mulheres, a idade média de 75.5 anos, sendo que 86.5 por cento deles tinham cuidadores. Entre os cuidadores, 80 por cento eram mulheres, membros da família, com idade média de 52.3 anos. Dedicavam 15.10 horas/dia para a atividade de cuidar, sem nenhuma ajuda. Encontrou-se importante relação entre a sobrecarga do cuidador, o esforço físico e emocional, e o estágio da demência. A sobrecarga emocional foi maior nos estágios iniciais e tardios da demência, mas a diferença foi estatisticamente não-significante. Os resultados atuais revelam a urgente necessidade de planejar estratégias de suporte formal e informal para cuidadores de idosos brasileiros com demência.

La finalidad de este estudio fue identificar características de los cuidadores de ancianos con demencia, tipos de demanda de cuidados y relacionar la demanda con el estadio de la demencia. Este estudio seccional y observacional fue efectuado en 2004 con 104 ancianos y 90 cuidadores en Ribeirão Preto/SP/Brasil, a través del instrumento OARS y un cuestionario para el cuidador. De los 104 ancianos, 66,3 por ciento eran mujeres, la edad media fue de 75,5 años y 86,5 por ciento del total tenían cuidadores. Entre los cuidadores, 80 por ciento eran mujeres, miembros de la familia, con edad media de 52,3 años. Dedicaban 15,10 horas/día para la actividad de cuidar, sin ayuda ninguna. Se encontró una importante relación entre la sobrecarga del cuidador, el esfuerzo físico y emocional y el estadio de la demencia. La sobrecarga emocional resultó mayor en los estadios iniciales y tardíos de la demencia, esta diferencia no fue estadísticamente significativa. Los resultados actuales revelan la necesidad urgente de planear estrategias de soporte formal e informal para cuidadores de ancianos brasileños con demencia.

The importance of serological assays in diagnosing acute pulmonary histoplasmosis

Histoplasmosis is a systemic mycosis caused by inhalation of Histoplasma capsulatum microconidia. The disease does not normally affect immunocompetent individuals after a single, transient inhalation exposure. However, longer exposure may cause chronic or disseminated acute pulmonary infection. Herein, we report the case of a 24-year-old immunocompetent patient, who presented fever, cough and dyspnea for one month. The chest radiography revealed interstitial infiltrate and diffuse micronodules. The patient reported having had close and prolonged contact with bats. Diagnosis was confirmed by positive double immunodifusion and immunoblotting assays. She was treated with ketoconazole (400 mg) and there was complete resolution of the disease.

Use of immunoblotting assay improves the sensitivity of paracoccidioidomycosis diagnosis

The purpose of this work was to evaluate two serological assays: double immunodiffusion (DI) and immunoblotting (IB) in immunodiagnosis of paracoccidioidomycosis (PCM). We evaluated by IB assay 23 sera samples from patients with clinical confirmation of PCM, all of them with negative DI results against culture filtrate from Paracoccidioides brasiliensis isolate 113. For IB, as well as for comparative DI assay, we employed soluble components of the cell wall outer surface (SCCWOS) from P. brasiliensis isolate 113 cultivated at 36°C in Fava-Neto's agar medium for 5 and 10 days. Among the 20 sera samples analyzed by DI, 13 (65%) were negative and 7 (35%) were positive against SCCWOS obtained on the 5th and 10th days. By IB assay, 95.4% and 100% of sera reacted against gp43 and gp70 present in SCCWOS from the 5th day and 95.6% recognized these fractions when evaluated against SCCWOS from the 10th day. Our results demonstrated that the use of an immunoenzymatic assay significantly improves the sensitivity of PCM immunodiagnosis and also suggests that at least two serological tests for antibody detection should be adopted in cases of questionable diagnosis.

Use of immunoblotting assay improves the sensitivity of paracoccidioidomycosis diagnosis

The purpose of this work was to evaluate two serological assays: double immunodiffusion (DI) and immunoblotting (IB) in immunodiagnosis of paracoccidioidomycosis (PCM). We evaluated by IB assay 23 sera samples from patients with clinical confirmation of PCM, all of them with negative DI results against culture filtrate from Paracoccidioides brasiliensis isolate 113. For IB, as well as for comparative DI assay, we employed soluble components of the cell wall outer surface (SCCWOS) from P. brasiliensis isolate 113 cultivated at 36°C in Fava-Neto's agar medium for 5 and 10 days. Among the 20 sera samples analyzed by DI, 13 (65 percent) were negative and 7 (35 percent) were positive against SCCWOS obtained on the 5th and 10th days. By IB assay, 95.4 percent and 100 percent of sera reacted against gp43 and gp70 present in SCCWOS from the 5th day and 95.6 percent recognized these fractions when evaluated against SCCWOS from the 10th day. Our results demonstrated that the use of an immunoenzymatic assay significantly improves the sensitivity of PCM immunodiagnosis and also suggests that at least two serological tests for antibody detection should be adopted in cases of questionable diagnosis.

Cytotoxicity effect of algal polysaccharides on HL60 cells.

The present study shows the cytotoxic effect of three different classes of algal polysaccharides on HL60 cells. Three galactofucans, fucoidan, and glucan were the polysaccharides utilized in this analysis. The parameters used for evaluating cell viability were [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] (MTT) reduction, protein content, and phosphatase activity. We demonstrated stimulation of phosphatase activity, MTT reduction, and protein content in relation to three types of galactofucans (1, 2, and 3) with different molecular weights (1600, 1200, and 360 kD). However, when HL60 cells were treated with galactofucan type 3, the total protein remained unchanged. Under the same experimental conditions, an expressed increase in the phosphatase activity was detected when galactofucan 3 was utilized. In relation to the mitochondrial function, the stimulation was higher in cells treated with galactofucan type 1. Fucoidan did not have a significant effect on MTT reduction, but protein content was decreased (IC50 around 30 microg/ml). Glucan also activated all the parameters that were analyzed, and this effect was more expressed in the phosphatase activity and in the protein content. This study provides new insights into the cytotoxic action of polysaccharides on HL60 cells and suggests for the first time the possible involvement of phosphatases in this process.

Biosynthesis of chondroitinase and hyaluronidase by different strains of Paracoccidioides brasiliensis.

The biosynthesis of chondroitinase and hyaluronidase by different isolates of Paracoccidioides brasiliensis was investigated in 20 strains isolated from patients (17 strains), a penguin (Pygocelis adeliae, one strain), an armadillo (Dasypus novemcinctus, one strain) and the environment (dog food, one strain). All the P. brasiliensis isolates studied had the ability to produce chondroitinase and hyaluronidase, although differences in colony morphology and enzyme production were detected among them. These results suggest that further investigations should be carried out in the clinical field in order to clarify the potential role of P. brasiliensis enzyme production in the pathogenesis of paracoccidioidomycosis.

Production of proteinase and phospholipase by Paracoccidioides brasiliensis.

We have investigated the production of proteinase and phospholipase by 20 different isolates of Paracoccidioides brasiliensis. Isolates were grown in Bacto-peptone, Dextrose, pH 5.5, agar slants, at 27 degrees C for 30 days, and cultures were transferred onto Petri dishes containing basis medium and bovine serum albumin fraction V and sterile egg yolk as substrates for enzyme production, and incubated at 27 degrees C. After 30 days net enzyme activity was visualized and quantitatively evaluated, measuring a ratio between colony diameter and diameter of the transparent (proteinase) or white (phospholipase) ring zone surrounding it. Results demonstrated that all isolates had the ability to produce proteinase and phospholipase, even though variability in enzyme production was noted among different isolates of P. brasiliensis.

Lung tissue mechanics in the early stages of induced paracoccidioidomycosis in rats

Pulmonary dysfunction represents the most important cause of death in patients with paracoccidioidomycosis (PBM). In order to investigate the functional changes of the lungs in the early stages of PBM, a model of benign disease was developed by intratracheal challenge of 12-week old isogenic Wistar rats with 1 x 10(6) yeast forms of Paracoccidioides brasiliensis. Animals were studied 30 and 60 days after infection, when fully developed granulomas were demonstrable in the lungs. Measurements of airway reistance, lung elastance and tissue hysteresis were made during sinusoidal deformations (100 breaths/min, tidal volume = 2 ml) with direct measurement of alveolar pressure using the alveolar capsule technique. Infection caused a significant increase in hysteresis (infected: 1.69, N = 13; control: 1.13, N=12,P = 0.024, ANOVA), with no alterations in airway resitance or lung elastance. Histopathological analysis revealed the presence of fully developed granulomas located in the axial compartment of the lung interstitial space. These results suggest that alterations of tissue mechanics represent an early event in experimental PBM.

Utilidade da oftalmoscopia na hipertensäo arterial sistêmica aguda / Ophthalmoscopy in acute blood hypertension

O objetivo deste foi avaliar a prevalência de alteraçöes oftalmoscópicas na crise hipertensiva e sua relaçäo com o tipo de crise e o nível de pressäo sanguínea. Neste estudo prospectivo, 111 pacientes foram examinados através da oftalmoscopia direta, entre junho/94 e junho/96. Seguindo-se a classificaçäo de Keith-Wagener-Barker, foi realizado estadiamento do fundo de olho para determinar a presença de retinopatia. Entre os 111 pacientes, retinopatia estava presente em 91 (82 p/cento), 88 (79,3 p/cento) foram urgências hipertensivas, 23 (20,7 p/cento) foram emergências. A admissäo hospitalar por emergência hipertensiva associou-se com maior severidade de retinopatia e o nível de pressäo arterial (p>0,05). Este estudo sugere que retinopatia é frequente em pacientes com crise hipertensiva. Näo foi encontrada relaçäo entre o nível pressórico sanguíneo determinado por esfingmomanometria e a retinopatia. Houve associaçäo da admissäo hospitalar por emergência hipertensiva com maior severidade de retinopatia

IgG, IgM and IgA antibody response for the diagnosis and follow-up of paracoccidioidomycosis: comparison of counterimmunoelectrophoresis and complement fixation.

IgG, IgM and IgA antibodies to GP43 (glycoprotein fraction of Paracoccidioides brasiliensis) were measured by ELISA in 63 samples from 23 patients with paracoccidioidomycosis before and twice after chemotherapy was started. Antibodies against P. brasiliensis were detected by indirect immunofluorescence (IF) (IgG, IgM and IgA isotypes), counterimmunoelectrophoresis (CIE) and complement fixation. Two control groups composed of 19 healthy individuals and 12 patients with other diseases (six with histoplasmosis, three with tuberculosis and three with other mycoses). The highest efficiency percentages were found with IgG and IgA-ELISA (100%), IgG-IF (96.2%), CIE (94.4%) and the lowest with CF (75.9%). Highest positive and negative predictive values (100%) were observed for IgG and IgA ELISA. IgG and IgM-ELISA antibodies are more often found in patients with acute than chronic disease (P = 0.01). Four to six months after treatment follow-up showed decreased levels of IgG and IgM-ELISA for acute cases and decreased titres of CIE for chronic cases in relation to pretreatment levels. This study suggests that IgG-ELISA anti-GP43 represents a good marker to monitor clinical response to therapy.

Monocyte adherence to Paracoccidioides brasiliensis, zymosan-C3b and erythrocyte-hemolysin in patients with paracoccidioidomycosis.

Independent and dependent (C3b/Fc receptors) opsonic adherence ability of monocytes from thirty-three patients with acute or chronic paracoccidioidomycosis and from 13 healthy individuals were studied in the presence of Paracoccidioides brasiliensis (Pb), Paracoccidioides brasiliensis opsonized by patient's serum (PbPS) or normal serum (PbNS), zymosan opsonized by fresh sera from healthy donors (ZyNS) and erythrocytes opsonized by hemolysin (EA). Statistically significant differences concerning the percentage of adhered monocytes to PbPS (number of adhered monocytes/total number of monocytes) were detected between control and chronic (active and inactive) groups. Significant differences in relationship to the mean number of PbPS (number of fungi in monocytes/total number of monocytes) were also observed between control and chronic active mycosis. Present data suggest that patients with chronic disease have more ability in the first step of phagocytic activity, considered as the main effector mechanism to control the dissemination and severity of paracoccidiodomycosis.

Lung tissue mechanics in the early stages of induced paracoccidioidomycosis in rats.

Pulmonary dysfunction represents the most important cause of death in patients with paracoccidioidomycosis (PBM). In order to investigate the functional changes of the lungs in the early stages of PBM, a model of benign disease was developed by intratracheal challenge of 12-week old isogenic Wistar rats with 1 x 10(6) yeast forms of Paracoccidioides brasiliensis. Animals were studied 30 and 60 days after infection, when fully developed granulomas were demonstrable in the lungs. Measurements of airway resistance, lung elastance and tissue hysteresis were made during sinusoidal deformations (100 breaths/min, tidal volume = 2 ml) with direct measurement of alveolar pressure using the alveolar capsule technique. Infection caused a significant increase in hysteresis (infected: 1.69, N = 13; control: 1.13, N = 12, P = 0.024, ANOVA), with no alterations in airway resistance or lung elastance. Histopathological analysis revealed the presence of fully developed granulomas located in the axial compartment of the lung interstitial space. These results suggest that alterations of tissue mechanics represent an early event in experimental PBM.

Immunochemical study of a Paracoccidioides brasiliensis polysaccharide-like antigen.

The polysaccharide antigen from P. brasiliensis has been largely employed in serologic tests ,as well as in skin tests, to evaluate cellular immunity. SDS-PAGE analysis of this antigen has revealed a variability in the number of bands exhibited by isolates SN, 265, 339, 113, and 18 (7 to 16 bands). The antigens obtained from isolates 2, PTL, 192 and Adel showed two or three bands. Glycoprotein analysis demonstrated a broad region between 50 and 90 kDa. Major bands of 48 and 30 kDa were present in almost all antigens. Optimal complement fixing dilution appears to be unaffected by the number of bands presented by different antigens. The immunoblot analysis revealed that the 90 and 30 kDa bands were mainly recognized by sera from paracoccidioidomycosis patients. Bands of high molecular weight were also recognized by most of the sera studied. Sera from histoplasmosis recognized the 94 kDa band. In conclusion, although the isolates exhibit quantitative variability in the number of fractions, it is possible to use only one or two samples given the greatest frequency of reactivity is seen in the 30 and 90 kDa fractions.

Responses of T and B lymphocytes to a Paracoccidioides brasiliensis cell wall extract in healthy sensitized and nonsensitized subjects.

Antigen-specific cellular immunity in paracoccidioidomycosis (PCM) has been poorly studied due to lack of standard in vitro lymphocyte proliferation assays. To standardize such an assay, we studied T and B cell responses to a Paracoccidioides brasiliensis cell wall extract (PbAg) in healthy subjects sensitized to either P. brasiliensis [Pb(+)Hc(-)] or to Histoplasma capsulatum [Hc(+)Pb(-)], and in nonsensitized persons. All subjects showed, as expected, a vigorous proliferative response to a control fungal antigen obtained from Candida albicans. Lymphocytes from Pb(+)Hc(-) donors, but not from Pb(-)Hc(-) donors, reacted to PbAg by proliferating in a dose-dependent manner with a maximum reaction after 6-9 days, suggesting a secondary specific immune response. Most activated cells were CD+CD4+ lymphocytes. However, Hc(+)Pb(-) donors' cells reacted with PbAg. Cross-reactivity with H. capsulatum was not unexpected, since both fungi, but not C. albicans, share cell wall immunogenic compounds. An enzyme-linked immunosorbent assay to detect human immunoglobulins (Ig) demonstrated that B cells from Pb(+)Hc(-) donors, but not from Pb(-)Hc(-) ones, reacted with PbAg by secreting high levels of IgG and IgM in 12-day culture supernatants. This secretion was possibly mediated by PbAg-activated CD4+ cells. We believe that analysis of T and B lymphocyte responses to PbAg will be useful in the investigation of the infection-associated immune impairment seen in some PCM patients.

Reacao enxerto x hospedeiro: forma cronica com acometimento pulmonar / Graft vs host reaction: chronic disease with pulmonary involvement

Os autores descrevem a evolucao de uma menina de 11 anos com reacao enxerto versus hospedeiro-forma cronica, pos transplante de medula ossea por anemia aplastica com acometimento pulmonar comprovado atraves de biopsia.

Lack of reactivity of paracoccidioidomycosis sera in the double immunodiffusion test with the gp43 antigen: report of two cases.

Sera from two patients with chronic active paracoccidioidomycosis yielded negative double immunodiffusion results with a culture filtrate antigen from Paracoccidioides brasiliensis routinely used in our laboratory. Complement fixation tests were positive for both sera using a polysaccharide-rich antigen. This study reports the results of a more extensive serological investigation of these two sera. Both a somatic antigen and a saline extract from the fungus yielded positive results in the double immunodiffusion. However, the immunodominant 43 kDa glycoprotein antigen showed negative results, although it was recognized by both sera in the Western blot assay. The value of the double immunodiffusion as a single serological test in paracoccidioidomycosis diagnosis is discussed.

Paracoccidioides brasiliensis. A mycologic and immunochemical study of a sample isolated from an armadillo (Dasipus novencinctus).

A sample of P. brasiliensis isolated from the spleen and the liver of an armadillo (Dasipus novencinctus) has been analysed under a mycological and immunochemical viewpoint. The armadillo was captured in an area of Tucuruí (State of Pará, Brazil), the animal being already established as an enzootic reservoir of P. brasiliensis at that region of the country. This sample maintained in the fungal collection of the Tropical Medicine Institute of São Paulo (Brazil) numbered 135, has got all the characteristics of P. brasiliensis, with a strong antigenic power and low virulence for guinea-pigs and Wistar rats. The specific exoantigen of P. brasiliensis--the glycoprotein with a molecular weight of 43 kDa--was easily demonstrated with double immunodiffusion, immunoelectrophoresis, SDS-PAGE and immunobloting techniques.