Genetic Diversity in Nannotrigona testaceicornis (Hymenoptera: Apidae) Aggregations in Southeastern Brazil.

The Meliponini, also known as stingless bees, are distributed in tropical and subtropical areas of the world and plays an essential role in pollinating many wild plants and crops These bees can build nests in cavities of trees or walls, underground or in associations with ants or termites; interestingly, these nests are sometimes found in aggregations. In order to assess the genetic diversity and structure in aggregates of Nannotrigona testaceicornis (Lepeletier), samples of this species were collected from six aggregations and genetically analyzed for eight specific microsatellite loci. We observed in this analysis that the mean genetic diversity value among aggregations was 0.354, and the mean expected and observed heterozygosity values was 0.414 and 0.283, respectively. The statistically significant Fis value indicated an observed heterozygosity lower than the expected heterozygosity in all loci studied resulting in high homozygosis level in these populations. In addition, the low number of private alleles observed reinforces the absence of structuring that is seen in the aggregates. These results can provide relevant information about genetic diversity in aggregations of N. testaceicornis and contribute to the management and conservation of these bees' species that are critical for the pollination process.

Small but not isolated: a population genetic survey of the tropical tree Cariniana estrellensis (Lecythidaceae) in a highly fragmented habitat.

Here, we explore the mating pattern and genetic structure of a tropical tree species, Cariniana estrellensis, in a small population in which progeny arrays (n=399), all adults (n=28) and all seedlings (n=39) were genotyped at nine highly informative microsatellite loci. From progeny arrays we were able to identify the source tree for at least 78% of pollination events. The gene immigration rates, mainly attributable to pollen, were high, varying from 23.5 to 53%. Although gene dispersal over long distance was observed, the effective gene dispersal distances within the small population were relatively short, with mean pollination distances varying from 69.9 to 146.9 m, and seed dispersal distances occurring up to a mean of 119.6 m. Mating system analyses showed that C. estrellensis is an allogamous species (tm=0.999), with both biparental inbreeding (tm-ts=-0.016) and selfing rates (s=0.001) that are not significantly different from zero. Even though the population is small, the presence of private alleles in both seedlings and progeny arrays and the elevated rates of gene immigration indicate that the C. estrellensis population is not genetically isolated. However, genetic diversity expressed by allelic richness was significantly lower in postfragmentation life stages. Although there was a loss of genetic diversity, indicating susceptibility of C. estrellensis to habitat fragmentation, no evidence of inbreeding or spatial genetic structure was observed across generations. Overall, C. estrellensis showed some resilience to negative genetic effects of habitat fragmentation, but conservation strategies are needed to preserve the remaining genetic diversity of this population.

Retrospective analysis of the use of G-CSF and its impact on dose response for anthracycline plus taxane-based schedules in early breast cancer

PURPOSE: T cells are dominant in the immune regulation of malignant pleural effusion (MPE). However, it is unclear about the role of IL-17+ T cells, particularly for IL-17+CD8+ Tc17 cells in antitumor immunity. This retrospective study is aimed at evaluating the prognostic significance of IL-17+ T cells in patients with MPE. METHODS: The frequency of IL-17+CD4+ Th17 and IL-17+CD8+ Tc17 cells in peripheral blood (PB), pleural fluids (PF), and tumor tissues in 24 patients undergoing thoracoscopy was determined by flow cytometry, immunohistochemistry, and ELISA. The association among the different measures was analyzed by Spearman's correlation tests. RESULTS: The percentages of PF Th17 and Tc17 cells were significantly higher than those in the PB of MPE patients and healthy controls (p < 0.01). Analysis of Th17 and Tc17 cells in the tumor tissues indicated that the percentages of Th17 and Tc17 cells in the invading tumor edge were significantly higher than those in the non-tumor tissues and intra-tumor regions (p < 0.05). More importantly, the percentages of IL-17+ T cells were associated with prolonged survival of patients with MPE. CONCLUSIONS: Both Th17 and Tc17 cells were involved in the tumor immunity against MPE. Increased frequency of Tc17 cells may serve as a biomarker for the prognosis of patients with MPE (AU)

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Retrospective analysis of the use of G-CSF and its impact on dose response for anthracycline plus taxane-based schedules in early breast cancer.

PURPOSE: To evaluate the impact on survival of the relative dose intensity (RDI) achieved in patients with early breast cancer receiving anthracycline plus taxane-based chemotherapy in the adjuvant setting. PATIENTS AND METHODS: Patients with early breast cancer diagnosed from January 1999 through December 2006 were included. Dose intensity was evaluated according to the number of delayed cycles and days and the percentage of RDI. RESULTS: A total of 231 breast cancer patients were included. Granulocyte colony-stimulating factor (G-CSF) was given to 39 % of patients. Few patients delayed chemotherapy administration ≥2 cycles (6 %) and ≥15 days (2 %), and the majority of them received ≥85 % of the RDI (98 %). Overall survival was statistically lower at 5 years in patients who received <85 % of RDI in comparison with those who received ≥85 % of RDI (80 vs. 97 %; p = 0.026). CONCLUSIONS: With a wide use of G-CSF in patients treated with adjuvant anthracyclines plus taxane-based schedules, 98 % of patients received a RDI ≥85 %. A significant although inconsistent impairment of survival was found in those patients with lower RDI.

Variabilidade genética em algumas criações comerciais brasileiras de escargots (Helix aspersa, Müller, 1774) / Genetic variation at eight isoenzyme loci in subpopulations of the edible snail (Helix aspersa, Müller, 1774)

Descreveram-se os marcadores isoenzimáticos e estimou-se a variabilidade genética de 20 subpopulações brasileiras de escargots (Helix aspersa). O estudo dos oito locos foi feito por eletroforese em gel de amido, em amostras com 30 indivíduos cada, obtidas em criatórios dos estados de Santa Catarina, São Paulo e Rio de Janeiro (uma, duas e 17 amostras, respectivamente). Observou-se polimorfismo nos locos das enzimas LAP, 6-PGD, PEP 2, PEP 1 e MDH, com três alelos nos três primeiros locos e dois nos demais. Os locos da ME, da SOD e da PGI apresentaram-se monomórficos. As freqüências gênicas de sete amostras ajustaram-se ao modelo de Hardy-Weinberg (P<0,05), e as de outras seis amostras ajustaram-se ao modelo de Wright (P<0,05), indicando que elas estão submetidas a diferentes regimes reprodutivos. Os desvios da panmixia para toda a população (F IT ) e dentro das subpopulações (F IS) não foram significativos (P³0,05). O desvio entre as subpopulações (F ST=0,0485) foi significativo (P<0,05) e apontou pequena diferenciação entre elas. As estimativas de diversidade total (Ht), entre subpopulações (Dst) e dentro das subpopulações (Hs), indicaram que a diversidade genética é reduzida e sua maior parte encontra-se dentro das subpopulações, sugerindo uma base genética estreita para essa população. As distâncias genéticas também foram pequenas, não permitindo a construção de um dendrograma.

Genetic structure of Blastocerus dichotomus populations in the Parana River basin (Brazil) based on protein variability.

The population structure of 147 marsh deer (Blastocerus dichotomus) from three areas in the Paraná River basin, Brazil, was studied by observing protein polymorphism at 17 loci. Six loci were polymorphic and 11 monomorphic. The proportion of polymorphic loci (P) was 35.29% and the average heterozygosity (H) was 6.31%. Wright's FST indicated that only 4.9% of the total variation in allelic frequencies was due to genetic differences between the three groups. The high value of F(IS) (0.246) indicated inbreeding in the marsh deer. Genetic distance values (D = 0.014-0.051) showed little divergence between the three areas. We suggest that probable mechanisms accounting for the genetic structure are female phylopatry and polygyny and also that inbreeding has resulted from decreasing areas of wetland leading to isolation, overhunting, and diseases transmitted by cattle.

Identification of microsatellite DNA markers for the giant anteater Myrmecophaga tridactyla.

The giant anteater (Myrmecophaga tridactyla) is found from Belize and Guatemala to Paraguay and Argentina. Its conservation status is considered vulnerable by IUCN. Here we report the isolation and characterization of six microsatellite loci. Positive loci for (GT)(n) were isolated using a magnetic bead hybridization selection protocol. The number of alleles per locus as well as the heterozygosity and PCR conditions are described. These loci will be useful for studying population structure, genetic diversity, and paternity in M. tridactyla wild populations.

Predominant African-derived mtDNA in Caribbean and Brazilian Creole cattle is also found in Spanish cattle (Bos taurus).

African-derived mitochondrial DNA (mtDNA) have been described in South American and Caribbean native cattle populations, which could have been introduced into America from Iberia or by direct importation from Africa. However, the similarity among described haplotypes is not known. We examined mtDNA variation in Guadeloupe Creole and Spanish cattle in an attempt to identify African-derived mtDNA haplotypes and compare them with those previously described. Eleven haplotypes clustered into the European taurine haplogroup (T3), two haplotypes into the African taurine (T1) haplogroup, and three haplotypes into the African-derived American haplogroup (AA). The AA1 and Eucons haplotypes were the most frequently observed. The presence of the AA haplogroup in Spanish cattle confirms historical records and genetic evidence of Iberian cattle as the main source of American native cattle origin. The possible origin of African-derived mitochondrial haplotypes in Iberian and Creole cattle is discussed, and the accumulated evidence does not support a founder effect from African ancestral cattle by direct importations. The presence of taurine AA and T3 haplotypes in Brazilian Nellore may indicate introgression by local European-derived cattle. Data presented in this work will contribute to the understanding of the origin of Guadeloupe Creole cattle.

African-derived mitochondria in South American native cattle breeds (Bos taurus): evidence of a new taurine mitochondrial lineage.

This article reports the nucleotide diversity within the control region of 42 mitochondrial chromosomes belonging to five South American native cattle breeds (Bos taurus). Analysis of these data in conjunction with B. taurus and B. indicus sequences from Africa, Europe, the Near East, India, and Japan allowed the recognition of eight new mitochondrial haplotypes and their relative positions in a phylogenetic network. The structure of genetic variation among different hypothetical groupings was tested through the molecular variance decomposition, which was best explained by haplotype group components. Haplotypes surveyed were classified as European-related and African-related. Unexpectedly, two haplotypes within the African cluster were more divergent from the African consensus than the latter from the European consensus. A neighbor-joining tree shows the position of two haplotypes compared to European/African mitochondrial lineage splitting. This different and putatively ancestral mitochondrial lineage (AA) is supported by the calibration of sequence divergence based on the Bos-Bison separation. The European/African mitochondria divergence might be subsequent (67,100 years before present) to that between AA and Africans (84,700 years before present), also preceding domestication times. These genetic data could reflect the haplotype distribution of Iberian cattle five centuries ago.

African dung beetle Onthophagus gazella Fabricius (Coleoptera: Scarabaeidae) esterase isozymes

African beetles Onthophagus gazella from both sexes were analyzed by electrophoresis for an investigation of esterase isozymes using alpha-naphthyl propionate and methylumbelliferyl propionate as substrates. Only one of the esterases (Est. 6) reacted with one of the substrates (alpha-naphthyl propionate). Six areas of activity were found, two of them being polymorphic (Est. 3 and Est. 4). For presence of Est. 3, 337 individuals were analyzed, including descendants of 32 controlled crossings: two alleles were identified, whose frequencies are Est. 3A = 0.447 and Est. 3B = 0.553. The population is in equilibrium for this locus (qui-square = 4.18; 0.2 > P > 0.1). For Est. 4, 338 individuals, descendants of 32 controlled crossings, were analysed. In this case, three alleles were identified whose frequencies are: Est. 4A = 0.277; Est. 4B = 0.661; and Est. 4C = 0.062. The population is not in equilibrium for this locus (qui-square = 40.259; p < 0.001). Two esterases were detected only in the pupal stage and another one in larvae. Of the 23 loci analyzed in these insects up to now, 3 are polymorphic (13 percent), which indicates very low variability in the population here studied

African dung beetle Onthophagus gazella Fabricius (Coleoptera: Scarabaeidae) esterase isozymes

African beetles Onthophagus gazella from both sexes were analyzed by electrophoresis for an investigation of esterase isozymes using alpha-naphthyl propionate and methylumbelliferyl propionate as substrates. Only one of the esterases (Est. 6) reacted with one of the substrates (alpha-naphthyl propionate). Six areas of activity were found, two of them being polymorphic (Est. 3 and Est. 4). For presence of Est. 3, 337 individuals were analyzed, including descendants of 32 controlled crossings: two alleles were identified, whose frequencies are Est. 3A = 0.447 and Est. 3B = 0.553. The population is in equilibrium for this locus (qui-square = 4.18; 0.2 > P > 0.1). For Est. 4, 338 individuals, descendants of 32 controlled crossings, were analysed. In this case, three alleles were identified whose frequencies are: Est. 4A = 0.277; Est. 4B = 0.661; and Est. 4C = 0.062. The population is not in equilibrium for this locus (qui-square = 40.259; p 0.001). Two esterases were detected only in the pupal stage and another one in larvae. Of the 23 loci analyzed in these insects up to now, 3 are polymorphic (13%), which indicates very low variability in the population here studied.

Foram analisados adultos de ambos os sexos do besouro africano Onthophagus gazella por eletroforese, para investigação de isozimas de esterases, utilizando como substratos alfa-naftil propionato e metil umbeliferil propionato. Somente uma das esterases (Est. 6) reagiu apenas com um dos substratos (alfa-naftil propionato). Foram encontradas seis regiões de atividades diferentes, sendo duas polimórficas (Est. 3 e Est. 4). Na região de Est. 3, foram analisados 337 indivíduos, incluindo descendentes de 32 cruzamentos controlados, e identificados 2 alelos, cujas freqüências são: Est. 3A = 0,447 e Est. 3B = 0,553. A população está em equilíbrio para esse loco (qui-quadrado = 4,18; 0,2 > p > 0,1). Na região de Est. 4, foram analisados 338 indivíduos, incluindo descendentes de 32 cruzamentos controlados, e identificados 3 alelos, cujas freqüências são: Est. 4A = 0,277; Est. 4B = 0,661; e Est. 4C = 0,062. A população não está em equilíbrio para esse loco (qui-quadrado = 40, 259; p 0,001). Foram detectadas duas regiões de esterases características da fase de pupa e uma região que aparece somente na larva. Do total de 23 locos amostrados até o momento nesses insetos, 3 são polimórficos (13%), o que indica variabilidade genética muito baixa na população estudada.

Effect of sub-MICs of antimycotics on expression of intracellular esterase of Trichophyton rubrum.

The electrophoretic pattern of the intracellular esterase of the dermatophyte Trichophyton rubrum was altered when this fungus was grown in the presence of subinhibitory concentrations of the antimycotics tioconazole or griseofulvin. All strains (original isolate and antimycotic resistant mutants) presented five clearly visible bands when cultivated on medium containing below-minimum inhibitory concentrations (sub-MICs) of tioconazole or griseofulvin, and only two clearly visible bands when cultivated in medium without antimycotics. No extra bands were detected in the electrophoretic patterns of the extracellular esterase of these fungi (mutants or the original isolate) when cultivated with or without tioconazole or griseofulvin (sub-MIC values). These results suggest that additional forms of esterase are produced inside the cell and may be a nonspecific response to cellular stress, or may participate in cellular detoxification processes in the presence of these antimycotics.

African dung beetle Onthophagus gazella Fabricius (Coleoptera: Scarabaeidae) esterase isozymes.

African beetles Onthophagus gazella from both sexes were analyzed by electrophoresis for an investigation of esterase isozymes using alpha-naphthyl propionate and methylumbelliferyl propionate as substrates. Only one of the esterases (Est. 6) reacted with one of the substrates (alpha-naphthyl propionate). Six areas of activity were found, two of them being polymorphic (Est. 3 and Est. 4). For presence of Est. 3, 337 individuals were analyzed, including descendants of 32 controlled crossings: two alleles were identified, whose frequencies are Est. 3A = 0.447 and Est. 3B = 0.553. The population is in equilibrium for this locus (qui-square = 4.18; 0.2 > P > 0.1). For Est. 4, 338 individuals, descendants of 32 controlled crossings, were analysed. In this case, three alleles were identified whose frequencies are: Est. 4A = 0.277; Est. 4B = 0.661; and Est. 4C = 0.062. The population is not in equilibrium for this locus (qui-square = 40.259; p < 0.001). Two esterases were detected only in the pupal stage and another one in larvae. Of the 23 loci analyzed in these insects up to now, 3 are polymorphic (13%), which indicates very low variability in the population here studied.

Restriction fragment length polymorphism (RFLP) in exon 2 of the BoLA-DRB3 gene in South American cattle.

The Bola-DRB3 gene participates in the development of the immune response and is highly polymorphic. For these reasons, it has been a candidate gene in studies of the genetic basis of disease resistance and in population genetic analysis. South American native cattle breeds have been widely replaced by improved exotic breeds leading to a loss of genetic resources. In particular, South American native breeds have high levels of fertility and disease resistance. This work describes genetic variability in the BoLA-DRB3 gene in native (Caracu, Pantaneiro, Argentinean Creole) and exotic (Holstein, Jersey, Nelore, Gir) cattle breeds in Brazil and Argentina. PCR-RFLP alleles were identified by combining the restriction patterns for the BoLA-DRB3.2 locus obtained with RsaI, BstY and HaeIII restriction enzymes. Allelic frequencies and deviations from the Hardy-Weinberg equilibrium were also calculated. Analysis of the 24 BoLA-DRB3 PCR-RFLP alleles identified showed differences in the allele distributions among breeds.

Structure and genetic relationship among Brazilian naturalized and imported goat breeds.

Fourteen goat populations were studied regarding their genetic relationship and structure. Parameters of genetic diversity (HT, HS and GST) and F statistic (FIS, FIT and FST) were estimated. Undefined breed populations presented high homogeneity, as did imported breed populations. Naturalized breed populations showed high differentiation. The genetic distances separating these 14 goat populations were calculated from gene frequency data for eight blood genetic markers (esterase D, phosphoglucomutase 1, carbonic anhydrase II, peptidase B, amylase, haemoglobin, transferrin, and protein X). Working with the genetic distance matrix of Nei corrected for small samples (DA), we constructed a dendrogram using the unweighted pair group method with arithmetic mean. DA values ranged from 0.0027 to 0.1518. The dendrogram divided the populations into two groups, one consisting of three populations of naturalized breeds, and another including the other populations (imported breeds, undefined breeds and some other naturalized breeds).

Occult cancer in patients with venous thromboembolism: which patients, which cancers.

We have previously demonstrated that patients with idiopathic venous thromboembolism (VTE) have a higher frequency of underlying cancer. Now we present a retrospective analysis of our 5-year experience with a series of 674 consecutive otherwise healthy patients, and a more restricted battery of diagnostic tests. Occult cancer was found in 15 patients during admission. The diagnostic tools which led to suspect occult cancer were: abdominal CT-scan (4 patients); high carcinoembryonic levels (2 patients); and high prostate-specific antigen levels (9 patients). Eight further patients were diagnosed of cancer after discharge. Cancer was more commonly found in patients with idiopathic VTE: 13/105 patients (12%) versus 10/569 patients (2%); p <0.01; O.R.: 7.9 (95% CI: 3.14-20.09). During the same period of time we diagnosed VTE in 147 patients with previously known cancer. When overall considered, VTE was the first sign of malignancy in most patients with prostatic and pancreatic carcinoma. On the contrary, most patients with breast, lung, uterine and brain cancers developed VTE as a terminal event of the disease. At variance with VTE patients and previously known cancer, most patients with occult malignancy were at an early stage. Further studies are needed to confirm whether patients with idiopathic VTE could benefit from screening for occult cancer. Meanwhile, our findings may serve as guidelines for physicians in this field.

Enzymatic variability in natural populations of Aedes aegypti (Diptera:Culicidae) from Brazil.

Yellowfever mosquitoes, Aedes aegypti (L), were collected from 3 towns located within 70 km of each other in the State of Sao Paulo to evaluate the protein variability of natural populations. Electrophoretic analysis permitted the identification of 6 loci responsible for the production of leucine aminopeptidase, 3 for esterases and malic enzyme, 2 for malate dehydrogenase, isocitrate dehydrogenase and alpha-glycerophosphate dehydrogenase, and 1 for phosphoglucomutase. For the 3 populations, the range for alleles per locus (1.63-2.03), the proportion of polymorphic loci (37.5-50.0%), and the expected mean heterozygosity (0.48-0.53) were greater than reported for other populations of A. aegypti. Although the 3 populations were similar genetically, the 1.8% differentiation was significant.

Biology of Amazonian anopheline. XX. Ontogeny of esterases, leucine aminopeptidase and alpha-glycerophosphate dehydrogenase in Anopheles (Nyssorhynchus) darlingi Root, 1926 (Diptera, Culicidae).

The esterases, leucine aminopeptidase and alpha-glycerophosphate dehydrogenase revealed modifications in gene expressions during the development of Anopheles darlingi. The esterases showed five activity bands, 1 and 2 being more deeply stained during the larval stages than in pupae or adults, esterases 3 and 4 more deeply stained in pupae and adults whereas esterase 5 was present throughout development. Leucine aminopeptidase showed five activity bands: LAP2 and LAP5 were characteristic of larvae, LAP3 was specific for pupae and adults, LAP4 was detected only in pupae, and LAP1 and LAP6 were detected in all stages. alpha-Glycerophosphate dehydrogenase presented one activity band on starch gel whose intensity increased with development. Two activity bands were detected on polyacrylamide gel (alpha-GPDH1 and alpha-GPDH2) in 4th-instar larvae (old pigmented larvae) and this activity increased with development.