Partial purification and characterization of lysine-ketoglutarate reductase in normal and opaque-2 maize endosperms

Lysine-ketoglutaratc reductase catalyzes the first step of lysine catabolism in maize (Zea mays L.) endosperm. The enzyme condenses L-lysine and α-ketoglutarate into saccharopine using NADPH as cofactor. It is endosperm-specific and has a temporal pattern of activity, increasing with the onset of kernel development, reaching a peak 20 to 25 days after pollination, and thereafter decreasing as the kernel approaches maturity. The enzyme was extracted from the developing maize endosperm and partially purified by ammonium-sulfate precipitation, anion-exchange chromatography on DEAE-cellulose, and affinity chromatography on Blue-Sepharose CL-6B. The preparation obtained from affinity chromatography was enriched 275-fold and had a specific activity of 411 nanomoles per minute per milligram protein. The native and denaturated enzyme is a 140 kilodalton protein as determined by polyacrylamide gel electrophoresis. The enzyme showed specificity for its substrates and was not inhibited by either aminoethyl-cysteine or glutamate. Steady-state product-inhibition studies revealed that saccharopine was a noncompetitive inhibitor with respect to α-ketoglutarate and a competitive inhibitor with respect to lysine. This is suggestive of a rapid equilibriumordered binding mechanism with a binding order of lysine, α-ketoglutarate, NADPH. The enzyme activity was investigated in two maize inbred lines with homozygous normal and opaque-2 endosperms. The pattern of lysine-ketoglutarate reductase activity is coordinated with the rate of zein accumulation during endosperm development. A coordinated regulation of enzyme activity and zein accumulation was observed in the opaque-2 endosperm as the activity and zein levels were two to three times lower than in the normal endosperm. Enzyme extracted from L1038 normal and opaque-2 20 days after pollination was partially purified by DEAE-cellulose chromatography. Both genotypes showed a similar elution pattern with a single activity peak eluted at approximately 0.2 molar KCL. The molecular weight and physical properties of the normal and opaque-2 enzymes were essentially the same. We suggest that the Opaque-2 gene, which is a transactivator of the 22 kilodalton zein genes, may be involved in the regulation of the lysine-ketoglutarate reductase gene in maize endosperm. In addition, the decreased reductase activity caused by the opaque-2 mutation may explain, at least in part, the elevated concentration of lysine found in the opaque-2 endosperm.

Control of parasitic infections among school children in the peri-urban area of Botucatu, Sao Paulo, Brazil

The prevalence of intestinal parasitosis was investigated in a primary school located in Rubiao Junior, a peri-urban district of Botucatu, Sao Paulo state, Brazil, in order to assess the effect of treatment and practical measures of prophylaxis in the control of parasitic infections among 7-to-18-year-old school children of a low socio-economic status. The first series of parasitological examinations included 219 school children, of which 123 (56.1%) were found to be infected with one or more parasite species. Eighty-four children carrying pathogenic parasites were submitted to various anti-parasitic treatment schedules. We re-evaluated 75 (89%) students after 4 to 6 months postchemotherapy. The results indicate that the combination of treatment with prophylactic measures has been successful in the control of parasitic infections, since reinfection rates were generally low (≤5.3%), except for Giardia lamblia infections (18.6%), and a marked reduction on the prevalence rates was observed with a significant percentage of cure (≤73.1%) in children infected with most parasite species. The reasons for the apparent failure in the control of infections caused by Hymenolepsis nana and Strongyloides stercoralis are discussed.

HIV attitudes and practices among professors in a Brazilian dental school.

Although there is considerable published research on Acquired Immunodeficiency Syndrome (AIDS), individual biases persist because of lack of information regarding HIV virus transmission. As a result, both infected patients and health care professionals suffer. The objective of this study was to determine if there is prejudice among university professors at the School of Dentistry at Aracatuba's Sao Paulo State University (FOA-UNESP) concerning HIV-positive patients or HIV-positive health care professionals. Out of the seventy-seven professors who responded to the questionnaire, 62.3 percent (forty-eight) stated that they advise their students not to refuse to treat a patient with HIV. Although 96.2 percent (fifty-two) of the fifty-four professors who treat patients have reported that they treat patients who are HIV-positive, only 65.3 percent of them were aware of infection control precautions, and only 32.7 percent reported that they would treat an HIV-positive patient like any other patient. There is also prejudice regarding HIV-positive professionals because only 48.1 percent (thirty-seven) of the professors responded that they would be willing to be treated by an infected professional. It can be concluded that there is prejudice among some of the FOA-UNESP university professors regarding individuals who are HIV-positive.

The relationship between onset of blood lactate accumulation, critical velocity, and maximal lactate steady state in soccer players

The objective of this study was to analyze the validity of the velocity corresponding to the onset of blood lactate accumulation (OBLA) and critical velocity (CV) to determine the maximal lactate steady state (MLSS) in soccer players. Twelve male soccer players (21.5 ± 1.0 years) performed an incremental treadmill test for the determination of OBLA. The velocity corresponding to OBLA (3.5 mM of blood lactate) was determined through linear interpolation. The subjects returned to the laboratory on 7 occasions for the determination of MLSS and CV. The MLSS was determined from 5 treadmill runs of up to 30-minute duration and defined as the highest velocity at which blood lactate did not increase by more than 1 mM between minutes 10 and 30 of the constant velocity runs. The CV was determined by 2 maximal running efforts of 1,500 and 3,000 m performed on a 400-m running track. The CV was calculated as the slope of the linear regression of distance run versus time. Analysis of variance revealed no significant differences between OBLA (13.6 ± 1.4 km·h-1) and MLSS (13.1 ± 1.2 km·h-1) and between OBLA and CV (14.4 ± 1.1 km·h-1). The CV was significantly higher than the MLSS. There was a significant correlation between MLSS and OBLA (r = 0.80), MLSS and CV (r = 0.90), and OBLA and CV (r = 0.80). We can conclude that the OBLA can be utilized in soccer players to estimate the MLSS. In this group of athletes, however, CV does not represent a sustainable steady-state exercise intensity. © 2005 National Strength & Conditioning Association.

Immune reconstitution in HIV-1 infected patients treated for two years with highly active antiretroviral therapy

The aim of this paper was to evaluate the immune reconstitution of HIV-1 patients subjected to highly active antiretroviral therapy (HAART) for two years or more according to CD 45RA and CD 45RO cell count; determination of IL-2, IFN-γ, IL-4, IL-10 and TNF-α serum levels; CD 4 + T and CD 8 + T lymphocyte count; and plasma viral load (VL) determination. For this purpose, a cross sectional study was carried out in the Tropical Diseases Area, Botucatu School of Medicine, São Paulo State University, UNESP, Botucatu, São Paulo, Brazil. Between June 2001 and April 2002, 37 HIV-1 infected patients were evaluated, 13 with treatment indication but untreated (G1), 9 subjected to HAART for 5-7 months (G2), and 15 treated for two years or more (G3); both treated groups used medication regularly and without failure. Forty-nine normal individuals were studied as controls (GC-1 and GC-2). There was a tendency (p<0.10) for the predominance of two nucleoside reverse transcriptase inhibitors (NRTI) associated with one non-nucleoside reverse transcriptase inhibitor (NNRTI) regimen in G2; and two NRTI associated with a protease inhibitor (PI) in G3. Statistical differences between groups were seen for CD 45RA (G1<[G3=GC-2]; p<0.05) and CD 45RO (G1[G2=G3]; p<0.001), TNF-α serum determination ([G1>G3; G2=intermediate]>GC-1; p<0.001), IL-2 (G1<[G2=G3=GC-1]; p<0.01), IFN-γ ([G1=GC-1]>[GC-2=G3]; p<0.001), IL-4 and IL-10 ([G1=G2=G3]>GC-1; p<0.001), serum cytokine profiles, with a higher proportion of subtype 2 in G1 and mature subtype 0 in G2 and G3 (p<0.005). There was no statistical difference for CD 8 + T lymphocyte counts (G1=G2=G3; p<0.50). Consistency was seen between positive correlations of profile 1 definer cytokines (IL-2 and IFN-γ), CD 45RA and CD 45RO cells, and CD 4 + T lymphocyte counts and between positive correlations of profile 2 definer cytokines (IL-4 and IL-10) with TNF-α, and VL. The negative correlations were also consistent as they expressed the inverse of the positives. The variables with the highest number of correlations were IL-2, IFN-γ, and VL, followed by CD 45RA and CD 45RO cells, and IL-10. The variables with the lowest number of correlations were CD 4 + T and CD 8 + T lymphocytes. The results express the partial but important immune reconstitution in HIV-1 infected individuals with the interference of HAART and the importance of cytokines especially IL-2 and IFN-γ, and CD 45RA and CD 45RO cells as surrogate markers of this reconstitution.

Serological profile of anti-Toxoplasma gondii antibodies in apparently healthy dogs of the city of Botucatu, São Paulo state, Brazil

Toxoplasmosis is a zoonosis caused by Toxoplasma gondii, a coccidian protozoan of worldwide distribution. The seroprevalence in canine population can be an alternative for measuring T. gondii urban spreading. A total of 780 blood samples from dogs were collected, during the yearly anti-rabies campaign, carried out by the Department of Veterinary Hygiene and Public Health, School of Veterinary Medicine and Animal Husbandry (FMVZ), São Paulo State University, UNESP, together with the county health authorities, in August 1999. Using Indirect Fluorescent Antibody Test (IFAT) for detecting antibodies anti-T. gondii in the sera samples, we observed that 258 dogs (33.1%) were positive. The associations between the serological results and the epidemiological variables were studied. Statistically significant differences were not found regarding sex (32.2% male and 34.3% female reactors). Dogs without a defined breed showed seropositivity statistically higher than the pedigreed group. The occurrence of infection was considered higher with age.

Occurrence of Actinobacillus actinomycetemcomitans in patients with chronic periodontitis, aggressive periodontitis, healthy subjects and children with gingivitis in two cities of the state of São Paulo, Brazil

The aim of this study was to determine the frequency of isolation of Actinobacillus actinomycetemcomitans (Aa) in 100 patients with chronic periodontitis, 14 patients with aggressive periodontitis, 142 pre-school children with gingivitis and 134 periodontally healthy subjects. Samples of subgingival plaque were taken using sterilized paper points introduced into periodontal pockets or gingival crevice for 60 seconds and inoculated on TSBV agar, which was incubated under anaerobiosis at 37°C, for 4 days. Microbial identification was performed through biochemical methods and morphocellular and morphocolonial analysis. Aa was detected in 40.3% of healthy subjects, 68% of patients with chronic periodontitis, 92.86% of patients with aggressive periodontitis and 40.14% of children with gingivitis. The rate of recovery of Aa in the tested human groups proved to be higher than previously reported and in agreement with participation of this facultative anaerobe as a member of native microbiota of the periodontium and its relation with aggressive and chronic periodontitis in Brazil.

Dental age in patients with Down syndrome

The aim of this research was to evaluate dental age in 102 patients with Down Syndrome, using panoramic radiographs. A software program developed by the Discipline of Radiology, School of Dentistry of São José dos Campos, São Paulo State University (UNESP), was used. A table of mineralization chronology of permanent teeth among Brazilians conceived by Nicodemo, Moraes and Medici Filho was used within the software. Statistical analysis of the results showed that 70.91% of the males land 61.21% of the females presented advanced dental age. Only 32.09% of the males and 39.79% of the females presented delayed dental age. Regarding the differences between the dental and chronological ages, two thirds of the males and females presented dental age with differences of up to 12 months, which means that they can be considered to be within normal standards, whereas only 18.87% of the males and 10.21% of the females presented dental age outside normal standards, with differences of over 24 months. In conclusion, the majority of the patients with Down Syndrome were considered to be within the normal standards of mineralization chronology.

Genetic data of 10 X-chromosomal loci in Vitória population (Espírito Santo State, Brazil)

Genetic population data for 10 X-STR (DXS8378, DXS9898, DXS7133, GATA31E08, GATA172D05, DXS7423, DXS6809, DXS7132, DXS9902 and DXS6789) were obtained from Vitória population (Espírito Santo State, Brazil). No deviations from the Hardy-Weinberg equilibrium and linkage disequilibrium were observed. The combined powers of discrimination in males and females were 0.9999995 and 0.99999999996, respectively. These high values show the potential of this system in human identification in Vitória population, Brazil. © 2009 Elsevier B.V. All rights reserved.

Aerobic fitness evaluation during walking tests identifies the maximal lactate steady state

Objective. The aim of this study was to verify the possibility of lactate minimum (LM) determination during a walking test and the validity of such LM protocol on predicting the maximal lactate steady-state (MLSS) intensity. Design. Eleven healthy subjects (24.2 ± 4.5 yr; 74.3 ± 7.7 kg; 176.9 ± 4.1 cm) performed LM tests on a treadmill, consisting of walking at 5.5 km h -1 and with 20-22% of inclination until voluntary exhaustion to induce metabolic acidosis. After 7 minutes of recovery the participants performed an incremental test starting at 7% incline with increments of 2% at each 3 minutes until exhaustion. A polynomial modeling approach (LMp) and a visual inspection (LMv) were used to identify the LM as the exercise intensity associated to the lowest [bLac] during the test. Participants also underwent to 24 constant intensity tests of 30 minutes to determine the MLSS intensity. Results. There were no differences among LMv (12.6 ± 1.7 %), LMp (13.1 ± 1.5 %), and MLSS (13.6 ± 2.1 %) and the Bland and Altman plots evidenced acceptable agreement between them. Conclusion. It was possible to identify the LM during walking tests with intensity imposed by treadmill inclination, and it seemed to be valid on identifying the exercise intensity associated to the MLSS. Copyright © 2012 Guilherme Morais Puga et al.

American visceral leishmaniasis: Factors associated with lethality in the state of São Paulo, Brazil

Objectives. To identify factors associated with death in visceral leishmaniasis (VL) cases. Patients and Methodology. We evaluated prognostic factors for death from VL in São Paulo state, Brazil, from 1999 to 2005. A prognostic study nested in a clinical cohort was carried out by data analysis of 376 medical files. A comparison between VL fatal cases and survivors was performed for clinical, laboratory, and biological features. Association between variables and death was assessed by univariate analysis, and the multiple logistic regression model was used to determine adjusted odds ratio for death, controlling confounding factors. Results. Data analysis identified 53 fatal cases out of 376 patients, between 1999 and 2005 in São Paulo state. Lethality was 14.1 (53/376), being higher in patients older than fifty years. The main causes of death were sepsis, bleeding, liver failure, and cardiotoxicity due to treatment. Variables significantly associated with death were severe anemia, bleeding, heart failure, jaundice, diarrhea, fever for more than sixty days, age older than fifty years, and antibiotic use. Conclusion. Educational health measures are needed for the general population and continuing education programs for health professionals working in the affected areas with the purpose of identifying and treating early cases, thus preventing the disease evolution towards death. © 2012 Geraldine Madalosso et al.

Oral health-related quality of life and satisfaction before and after treatment with complete dentures in a Dental School in Brazil

Purpose: The aim of this study was to evaluate the impact of treatment with new complete dentures on oral health-related quality of life and satisfaction of edentulous patients attending the Prosthodontic Department in a Dental School in Brazil. Methods: A total of 70 edentulous subjects (37-86 years) treated by undergradute students at the Araçatuba Dental School, University of State of São Paulo, were accessed. A specific questionnaire for edentulous patients (EDENT) based on the Oral Health Impact Profile (OHIP-EDENT) was applied to collect information on patient oral health-related quality of life. Questions related to the personal satisfaction of the edentulous patients with their complete dentures were also included. The patients were accessed before the treatment, and 3 months after receiving the new dentures. Results: After rehabilitation with new complete dentures, all domains of OHIP-EDENT showed significant improvements. There was also a significant improvement in patient satisfaction after placement of new complete dentures. Furthermore, it was possible to observe association between upper denture satisfaction and age. Conclusion: Conventional complete dentures may have a positive impact on oral health-related quality of life and satisfaction of edentulous patients. © 2012 Japan Prosthodontic Society.

Motor intervention in children with school learning difficulties

This study sought to evaluate motor development in children aged 6 to 11 years with learning difficulties and school characteristics of delayed motor development, before and after application of a motor intervention program. The sample consisted of 28 children with a mean age of 107.21 ± 16.56 months, who were evaluated by the Motor Development Scale and received motor intervention for 6 months, followed by reassessment. We observed a statistically significant difference between the average of the motor activity ratios in all areas of the evaluation and reevaluation. Also verified in the evaluation were the concentration ratios of children with motor activity greater than or equal to 80 and there was a revaluation increase in this concentration on re-evaluation, the areas with the greatest increase in concentration and significant differences being: Body Schema, Space and temporal Organization. In the overall evaluation of MDS, most children presented the classification of low normal. However, in the reassessment most have evolved into the average normal, only 4 of themremaining in the same classification. Therefore, in this study, children with learning disabilities also showed motor deficits and the intervention applied contributed to an increase in the motor ratios with consequent improvement in motor development. Besides psychopedagogical asistance, it is essential to reassess them and if necessary apply the intervention in the motor development of children with learning difficulties.

Avaliação dos níveis de citocinas e HLA-G solúvel em linhagens celulares tumorais de colo uterino tratadas com alcalóides de Pterogyne nitens

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

A prática pedagógica de professores do ensino fundamental na perspectiva de uma educação de qualidade para todos

Pós-graduação em Educação - FCT