Juarez Gabardo

New evidence that the MHC influences odor perception in humans: a study with 58 Southern Brazilian students

Increasing evidence suggests a correlation between mate choice, odor preference, and genetic similarity at the Major Histocompatibility Complex (MHC) in a variety of animals, including our species. The MHC is a highly polymorphic group of genes that play an important role in the immunological self/nonself recognition. Its products have been reported to take part on the variety of compounds and reactions that together build an individuals body odor. It has been suggested, therefore, that animals use body odor as a guide to identify possible mates as MHC-similar or MHC-dissimilar from their own genotype. Preference for a MHC-dissimilar partner enhances MHC heterozygosity of an individuals offspring. The possible adaptive advantages are clear: it is a mechanism of avoiding inbreeding and MHC-heterozygous offspring may have enhanced immunocompetence. The aim of this study was to search, in our species, new evidence on the correlation between specificities at HLA-A and HLA-B and assessments of pleasantness regarding specific body odors. HLA is the name for the human MHC. Four olfactory sessions were performed with 58 young Southern Brazilian students, in order to investigate whether assessments of pleasantness of body odors from individuals correlate to a persons HLA phenotype. Body odors were collected via sweat and urine from all participants. Women smelled and scored all male odor samples and men did the same with all female samples. We found a significant correlation only when female smellers evaluated male sweat odors.

Identification and colonization of endophytic fungi from soybean (Glycine max (L.) Merril) under different environmental conditions

ABSTRACT A total of 297 endophytic fungi were isolated from 1728 leaf and stem fragments collected about twenty and forty days after germination from soybean (Glycine max (L.) Merril) plants grown in the field and a greenhouse. The fungi belonged to eight groups, six dematiaceous genera (Alternaria , Cladosporium , Chaetomium , Curvularia , Drechslera and Scopulariopsis ) and the non-dematiaceous genera Acremonium , Aspergillus , Colletotrichum , Fusarium , Paecilomyces and Penicillium along with some Mycelia sterilia .. There were qualitative and quantitative differences in the type and number of isolates obtained from greenhouse and field-grown plants, with more isolates being obtained from the latter. No difference was found in the number of fungi isolated from leaves and stems irrespective of where the plants was grown. For was field-grown plants, the number of isolates decreased as the plants aged and more fungi were found in tissues near the soil, while for greenhouse-grown plants the number of isolates increased as the plants aged but in this case no more fungi were isolated from those tissues nearer the soil. These results could have biotechnological relevance for the biological control of pests or plant growth promotion . Key words: Endophytic fungi, Soybean, Biological control

Vaccinating against HPV: Physicians’ and medical students’ point of view

The present survey was conducted among medical students and physicians affiliated to the Federal University of Paraná, Curitiba, Brazil, between August 2006 and December 2007. 252 individuals responded to a questionnaire composed of 13 items regarding their individual status and their personal knowledge of the HPV vaccine properties. The data analysis that was carried out using chi-square test showed that 79.7% of the interviewed population would indicate the vaccine, and mostly, to girls aged 10-15 years old. While vaccine effectiveness and prophylaxis appeared to have been adequately understood, some of its properties such as safety and immunity duration still need further elucidation.

Establishment of experimental conditions for preserving samples of fish blood for analysis with both comet assay and flow cytometry

When environmental analysis is performed, the high number of samples required and handling conditions during the transport of these samples to the laboratory are common problems. The comet assay is a useful, highly sensitive tool in biomonitoring. Some studies in the literature aim to preserve slides in lysis solution for use in the comet assay. Until now, however, no efficient methodology for preserving blood samples for this assay has been described. Because of this, the present report aimed to establish the proper conditions for samples maintenance prior to comet assay analysis. Samples were conserved in three different solutions: a high protein concentration solution (fetal bovine serum-FBS), an anticoagulant agent (a calcium chelator - ethylenediaminetetracetic acid - EDTA), and a salt buffered solution (phosphate buffered saline-PBS). Therefore, peripheral blood samples of Rhamdia quelen specimens were collected and maintained in these solutions until testing at 72h. Analyses of DNA fragmentation via the comet assay and cell viability via flow cytometry were performed at intervals of 24h. The results showed that samples maintained in FBS were preserved better; this was followed by those preserved in PBS and then last by those preserved in EDTA. In conclusion, blood samples from freshwater fish can be preserved up to 48h in fetal bovine serum at 4 degrees C in the absence of light. In this period, no DNA fragmentation occurs. We thus describe an excellent method of sample conservation for subsequent analysis in the laboratory.

Metais pesados em solos de área de mineração e metalurgia de chumbo: I - Fitoextração

A fitoextracao tem sido sugerida como alternativa viavel as praticas tradicionais de recuperacao de solos contaminados por metais pesados (remocao do solo e destinacao em aterros ou coprocessamento em cimento, etc.), em razao dos menores custos e por ser menos impactante ao ambiente. Este trabalho objetivou avaliar a fitoextracao induzida (uso de acido citrico como agente quelante) de metais pesados, com o cultivo de aveia-preta (Avena strigosa Schreber), girassol (Helianthus annuus L.) e grama-batatais (Paspalum notatum Flugge), em solos poluidos (solos 2, 3 e 4) de uma area de mineracao de Pb em Adrianopolis (PR). O solo 1 foi amostrado em area de mata nativa (referencia). O experimento foi realizado em casa de vegetacao, com tres repeticoes. Foram determinados os teores de Pb, Cd, Cu, Ni, Cr e Zn no solo com HNO3/HCl (3:1) concentrados. Apos a colheita das plantas, determinaram-se a massa de materia seca e os teores de metais pesados nas raizes e parte aerea (digestao nitrico-perclorica). Os solos contaminados (2, 3 e 4) apresentaram as seguintes faixas de teores de metais pesados (mg kg-1): Pb - 2.598,5 a 9.678,2; Cd - 1,9 a 22,2; Cu - 165,5 a 969,2; Ni - 22,6 a 38,4; Cr - 15,2 a 27,8; e Zn - 87,4 a 894,8. A adicao de quelante nao induziu a uma absorcao mais efetiva de metais pesados pelas plantas. O solo 2 possibilitou melhor crescimento das plantas, e o girassol deve ser preferido na fitorremediacao das areas sob as mesmas condicoes. Na area proxima a planta fabril (solo 3) e nas areas com grande ocorrencia de rejeitos (solo 4), a fitoextracao nao foi eficiente. Nesses ambientes, recomenda-se o estudo de outras plantas nativas e, ou, exoticas resistentes a altos teores de Pb (solo 3) ou a remobilizacao de solo mais rejeito para aterros industriais (solo 4).

Activation of bone marrow cells treated with Canova in vitro

Canova is a Brazilian complex homeopathic medication produced from Aconitum, Thuya, Bryonia, Lachesis and Arsenicum. Previous studies demonstrated that Canova induces up‐regulation in numbers of leukocytes. The bone marrow microenvironment is composed of growth factors, stromal cells, extracellular matrix, and progenitor cells that differentiate into mature blood cells. As it is the major site of blood cell formation, we studied in vitro Canova effects on bone marrow cells of mice. Swiss mouse femurs were dissected, cleaned, and the marrow was flushed. The cells were plated, treated or not, incubated for different times and processed for light, scanning electron, and confocal microscopy, and also flow cytometry. The treatment did not modify the expression of the analyzed surface markers or cytokine production. All microscopy techniques showed that a monocytic lineage (CD11b+) and stromal cells (adherent cells) were activated by treatment. Canova also increased cell clusters over adherent cells, suggesting proliferation areas.

Stimulation of lymphocyte anti-melanoma activity by co-cultured macrophages activated by complex homeopathic medication

BackgroundMelanoma is the most aggressive form of skin cancer, and the most rapidly expanding cancer in terms of worldwide incidence. Chemotherapeutic approaches to treat melanoma have been uniformly disappointing. A Brazilian complex homeopathic medication (CHM), used as an immune modulator, has been recommended for patients with depressed immune systems. Previous studies in mice have demonstrated that the CHM activates macrophages, induces an increase in the number of leukocytes and improves the murine response against Sarcoma-180.MethodsHere we studied the interaction of mouse lymph node lymphocytes, co-cultured in vitro with macrophages in the presence or absence of the CHM, with B16F10 melanoma cells.ResultsLymphocytes co-cultured with macrophages in the presence of the CHM had greater anti-melanoma activity, reducing melanoma cell density and increasing the number of lysed tumor cells. There was also a higher proportion of activated (CD25+) lymphocytes with increased viability. Overall, lymphocytes activated by treatment destroyed growing cancer cells more effectively than control lymphocytes.ConclusionCo-culture of macrophages with lymphocytes in the presence of the CHM enhanced the anti-cancer performance of lymphocytes against a very aggressive lineage of melanoma cells. These results suggest that non-toxic therapies using CHMs are a promising alternative approach to the treatment of melanomas. In addition, they are attractive combination-therapy candidates, which may enhance the efficacy of conventional medicines by improving the immune response against tumor cells.

Metais pesados em solos de área de mineração e metalurgia de chumbo: II - formas e disponibilidade para plantas

The forms and availability of heavy metals in contaminated soils determine the uptake potential of plants and water contamination by leaching. In this study different methods of chemical extraction were used, in order to identify the forms of Pb, Cd, Cr, Ni, Cu, and Zn and evaluate the availability of these pollutants for sunflower (Helianthus annuus L.), black oat (Avena strigosa Schreber) (exotic species) and Bahiagrass (Paspalum notatum Flugge) (native species), as indicators, in soils of Pb mining and metallurgy in Adrianopolis, state of Parana, Brazil. Soil samples were collected in the 0-40 cm layer in four environments of the area: soil 1 - control (native forest); soil 2 - waste in soil profile; soil 3 - near the factory chimney (deposition of particulate material); soil 4 - heavy deposition of waste. The following extraction methods were used: DTPA-TEA pH 7.3; Ca(NO3)2 0.5 mol L-1; HNO3 0.5, 1.0 and 4.0 mol L-1, and concentrated HNO3/HCl (3:1). The plants were grown in a greenhouse, with three replications. The increase of pseudo-total, exchangeable and non-exchangeable heavy metal contents was verified; the highest contents of Pb and Zn were extracted by HNO3/HCl; 9,678.2 and 894.8 mg kg-1, respectively. Soil heavy metals based on acid extractors HNO3 0.5, 1.0 and 4.0 mol L-1 and HNO3/HCl were strongly correlated with each other. The extractor based on chelation (DTPA-TEA pH 7.3) was not efficient to predict the availability of heavy metals for plants under the study conditions. The nitric extractions should be preferred to determine Pb and Zn plant availability in the area.

Fungicide resistance and genetic variability in plant pathogenic strains of Guignardia citricarpa

A Mancha Preta dos Citros (MPC) tem ocorrencia mundial afetando a producao de citros na Africa, Oceania e America do Sul. No Brasil, onde o clima e favoravel ao seu desenvolvimento, a doenca esta espalhada nas regioes Sul e Sudeste. O controle da MPC, causada pelo fungo Guignardia citricarpa (anamorfo: Phyllosticta citricarpa) e baseado na aplicacao de fungicidas, como os benzimidazois. Na Africa do Sul, apos 10 anos de controle da doenca com o fungicida benomil, os casos de resistencia a altas concentracoes deste fungicida atingiram todas as areas produtoras. O fungicida estrolilurina chamado azoxistrobina tem se mostrado eficiente no controle dos fitopatogenos de uma grande variedade de culturas economicamente importantes, incluindo a MPC. Neste trabalho foram investigados os efeitos in vitro dos fungicidas benomil e azoxistrobina em 10 linhagens de G. citricarpa isoladas de lesoes em plantas citricas no Brasil e na Africa do Sul. Houve inibicao do crescimento micelial a 0,5 µg/mL do fungicida benomil entre as linhagens testadas, com excecao de PC3C de origem sul-africana, que apresentou resistencia ate a concentracao de 100,0 µg/mL de benomil. A frequencia de mutacao espontânea para resistencia ao benomil foi de 1,25 ´ 10-7. A estrobilurina azoxistrobina, mesmo em altas concentracoes, nao inibiu o crescimento micelial dos isolados, entretanto reduziu significativamente a producao de esporos, chegando a 100% de inibicao em concentracoes de 5,0 µg/mL de azoxistrobina. A variacao na sensibilidade das linhagens, principalmente com a estrobilurina azoxistrobina, possivelmente esta relacionada com a variabilidade genetica dos isolados de G. citricarpa.

Evaluation of the protective potential of a Taenia solium cysticercus mimotope on murine cysticercosis

An NC-1 mimotope from Taenia solium cysticerci can help identify patients with neurocysticercosis through immunoassay. After chemical synthesis, an NC-1 peptide was coupled to bovine serum albumin (NC-1/BSA) for used as an immunogen in murine Taenia crassiceps cysticercosis, which is an experimental model of cysticercosis caused by T. solium. NC-1/BSA immunisation decreased parasitaemia by inducing 74% protection compared to the 77% protection obtained with T. crassiceps crude antigen. The influence of immunisation was also observed on the size and stage of development of the parasite. Antibodies from NC-1/BSA-immunised mice recognised proteins from the tegument and from the buddings, and intense immunostaining was observed in the final stage of the metacestode. The capacity of NC-1/BSA to induce protective antibodies which are reactive to proteins from the tegument of the metacestode suggests that this mimotope is a potential candidate for a vaccine against human and animal cysticercosis.