Effects of 17ß-estradiol on early gonadal development and expression of genes implicated in sexual differentiation of a South American teleost, Astyanax altiparanae.

Gonadal sex differentiation in teleost fish shows greater plasticity as compared to other vertebrates, as it can be influenced by a variety of factors such as exogenous sex steroids. Exogenous estrogens, such as 17ß-estradiol (E2), can induce feminization when administered during early embryonic development. However, the mechanisms underlying the E2-induced feminization are not fully understood, especially in Neotropical species. Therefore, the aim of this study was to evaluate the effects of E2 administration on the phenotypic sex characteristics, histological assessment of the gonads, and the expression of selected genes in Astyanax altiparanae exposed to dietary E2 prior to gonadal differentiation. At 4 days post-hatch (dph), groups of 30-40 undifferentiated larvae were fed with a diet containing varying amounts of E2 for 28 days, and fish were sampled at 90 dph. Previous studies revealed that ovary formation in A. altiparanae occurred at 58 dph, whereas the first sign of testis formation was found at 73 dph. In relation to the control, E2 exposure increased the proportion of phenotypic females in 120% and 148.4% for 4 and 6 mg E2/Kg, respectively. However, histological analysis revealed that treatments did not affect gonadal sex ratio between males and females, but induced intersex (testis-ova) in the group treated with 6 mg E2/Kg food. Treatment with E2 also altered gonadal transcript levels of a selected number of genes implicated in sexual differentiation. Males overexpressed dmrt1, sox9 and amh following E2 treatment as compared to control. Females showed increased mRNA levels of dmrt1 and sox9, which might be related to the down-regulation of cyp19a1a after E2 exposure. In summary, E2 exposure during early gonadal development affected male secondary characteristics without changing the gonadal sex ratio, and altered expression of genes implicated in sexual differentiation.

Distribution of constitutive heterochromatin in Pachycoris torridus (Hemiptera, Scutelleridae) with different chromatic patterns.

The stink bug Pachycoris torridus is a pest of great agricultural importance due to its records on culture of physic nut (Jatropha curcas), which is the raw material for biodiesel production. An interesting feature of this insect is its high phenotypic variability, a characteristic that resulted in it being classified as a new species on eight separate occasions. In the suborder Heteroptera, the heterochromatin pattern is specific and often allows species to be differentiated. To confirm whether there is differentiation between specimens of P. torridus with different color patterns (yellow, orange, brown, and red), samples were analyzed cytogenetically using the C-banding method. During meiotic prophase, the four color patterns analyzed showed a large heterochromatic chromocenter, consisting of a combination of both sex chromosomes (X and Y). Thus, the present study reports chromosomal homogeneity in different color patterns of P. torridus and highlights the importance of this tool in the description of new species.

High genetic variability and polychromatism in Pachycoris torridus (Heteroptera: Scutelleridae).

The stink bug Pachycoris torridus is listed among the most polyphagous insects in the world and it is a major pest of diverse crops, in particular the physic nut Jatropha curcas, which is used as a raw material for biodiesel production. A peculiar characteristic of this species is its high phenotypic variability, a characteristic that makes identification difficult: P. torridus has been described as a new species eight times. Thus, to aid in identification, genetic characterization of this insect was performed. We verified that, due to the high genetic variability of P. torridus, several genetic patterns exist that result in the same phenotype.

Study of nucleolar behavior during spermatogenesis in Martarega brasiliensis (Heteroptera, Notonectidae).

Few cytogenetic studies have been undertaken using aquatic heteropterans and the nucleolar behavior of these insects has been described in only four species, Limnogonus aduncus, Brachymetra albinerva, Halobatopsis platensis, and Cylindrostethus palmaris. The nucleolus is a cellular structure related to biosynthetic activity and it exhibits a peculiar behavior in the heteropterans of the Triatominae subfamily; it persists during all stages of meiosis. Thus, this study aims to analyze spermatogenesis in Martarega brasiliensis, with an emphasis on nucleolar behavior. Twenty M. brasiliensis adult males were used and collected from the Municipal reservoir in the city of São José do Rio Preto, São Paulo, Brazil. The species were fixed in methanol:acetic acid (3:1), then dissected, and the testicles were extracted, torn apart, and impregnated with silver ions. During prophase, the nuclei of M. brasiliensis were composed of the nucleolus and nucleolar corpuscles, which varied in number from one to four, emphasizing that this insect has great synthetic activity during meiosis. The analysis of cells in metaphase I showed that M. brasiliensis presents a nucleolar organizing region in at least one autosome. Furthermore, the phenomenon of nucleolar persistence was not observed. All spermatids presented nucleolar markings that varied in number and position according to the stage of elongation. Moreover, it was also possible to highlight the presence of a vesicle in spermatids. Thus, this paper describes the nucleolar behavior of M. brasiliensis and highlights important characteristics during spermatogenesis, thus, increasing the knowledge about the biology of these aquatic heteropterans.

Adaptação e adaptação cruzada de Listeria monocytogenes aos compostos eugenol e carvacrol / Adaptation and Cross adaptation of Listeria monocytogenes to eugenol and carvacrol compounds

RESUMOA evolução de certos microrganismos permite sua rápida adaptação aos ambientes em constante mudança, desenvolvendo assim, tolerância ou resistência ao aumento de determinados estresses. O uso de compostos bioativos provenientes da flora nativa tem sido apontado como uma possível solução para os problemas de controle da resistência e proliferação bacteriana. Este trabalho visou verificar a adaptação e adaptação cruzada de L. monocytogenes, frente aos compostos fenólicos eugenol e carvacrol. A concentração mínima inibitória (CMI) dos compostos fenólicos foi determinada pela técnica de microdiluição em placas de 96 cavidades, em caldo TSB + 0,5% de Tween 80. As concentrações finais (%) obtidas foram: 0,06; 0,12; 0,24; 0,49; 0,98; 1,95; 3,9; 6,25; 12,5; 25; 50. A suspensão bacteriana padronizada foi inoculada nas cavidades das placas, as quais foram incubadas a 37°C por 24 horas com posterior leitura da absorbância a 620 nm e determinação da CMI. A adaptação das células de L. monocytogenes ao eugenol e carvacrol foi realizada com o cultivo das células em TSB + 0,06% de eugenol ou carvacrol à 37°C por 2 horas. A cultura foi então centrifugada e as células ressuspendidas e padronizadas em TSB. A seguir, realizou-se novamente a técnica de microdiluição em caldo. Os resultados obtidos demonstraram que L. monocytogenes apresentou adaptação e adaptação cruzada frente ao carvacrol e eugenol. A CMI do eugenol e carvacrol foi de 24%. A pré-exposição de L. monocutogenes a concentração sub-letal de 0,06% de carvacrol ou de eugenol aumentou sua resistência. A pré-exposição ao carvacrol promoveu a adaptação de L.monocytogenes a ele aumentando a CMI para 12,5%. Já para o eugenol a CMI passou para 25%. Quando submetidas à concentração sub-letal de eugenol, este promoveu a adaptação das células tanto ao carvacrol quanto ao eugenol, sendo a CMI de 12,5%. Os resultados obtidos demonstraram que L. monocytogenes apresentou adaptação e adaptação cruzada ao carvacrol e eugenol. O presente trabalho sugere estudos futuros ainda mais abrangentes quanto à potencialidade antimicrobiana destes compostos.

ABSTRACTSome microorganisms have evolved and therefore are able to rapidly adapt themselves to a constantly changing environment, thus developing tolerance or resistance to the increase of some specific stresses. The use of bioactive compounds from the native vegetation has been pointed out as a possible solution to the problems of control of bacterial resistance and proliferation. This work aimed to check the adaptation and cross adaptation of L. monocytogenes, towards the phenolic compounds eugenol and carvacrol. The minimum inhibitory concentration (MIC) of the phenolic compounds was determined by the microdilution technic in plates of 96 cavities, in CALDO TSB + 0,5% of Tween 80. The final concentrations (%) obtained were: 0,06; 0,12; 0,24; 0,49; 0,98; 1,95; 3,9; 6,25; 12,5; 25; 50.The patronized bacterial suspension was inoculated into the captivities of the plates, and incubated at 37°C for 24 hours with posterior reading of the absorbance at 620 nm and determination of the MIC. The adaptation of the cells of L. monocytogenes to the eugenol and carvacrol was made through the cultivation of the cells in TSB + 0,06% of eugenol or carvacrol at 37°C for 2 hours. The sample was then centrifuged and the cells were re-suspended and patronized in TSB. After that, the microdilution technic was performed one more time. The obtained results revealed that the L. monocytogenespresented adaptation and cross adaptation to the eugenol and carvacrol. The eugenol and carvacrol CMI was at 24%. The pre-exposition of L. monocytogenes to sub-lethal doses of 0,06% of eugenol or carvacrol enhanced its resistance. The pre-exposition to carvacrol promoted the adaptation of L. monocytogenes to it thus increasingthe MIC to 12,5%. To the eugenol the CMI got to 25%. When submitted to sub-lethal concentrations of eugenol, the latterpromoted the adaptation of the cells to carvacrol and eugenol, bringing the CMI to 12,5%. The obtained results showed that theL. monocytogenespresented adaptation and cross adaptation to the eugenol and carvacrol. The current work suggests future studies even broader regarding the antimicrobial potentialityof these compounds.

Capim-piatã adubado com fontes de fósforo de diferente solubilidade em água / Piata palisadegrass fertilized with different sources of phosphorus for solubility in water

The development of piatã grass fertilized with phosphorus sources with different solubilities in water was evaluated. The experiment was developed in the unit of Aquidauana of UEMS, MS, from February to August, 2012. Four treatments were evaluated: control (without phosphorus fertilization), slow solubility, fast solubility (readily available), and mixed solubility sources of phosphorus (FH Pasture(r)). It was adopted a randomized blocks design with four blocks and four replicates in a total of 16 pastures (0.5 ha each one). Measurements were performed weekly to evaluate structural characteristics and morphogenic variables of the plants. Data were submitted to a variance analysis according to the model. When necessary, the means of treatments with different phosphorus fertilizer sources were compared with the control using the Dunnett test and with other treatments by using student's t test. The significance level of 5% was adopted in all analysis. No significant effect was observed (P 0.05) in the evaluated traits in the first year of the experiment.

Structural investigation and photoluminescent properties of gadolinium(III), europium(III) and terbium(III) 3-mercaptopropionate complexes.

This work reports on the synthesis, crystallographic determination and spectroscopic characterization of gadolinium(III), terbium(III) and europium(III) 3-mercaptopropionate complexes, aqua-tris(3-mercaptopropionate)lanthanide(III)--[Ln(mpa)3(H2O)]. The Judd-Ofelt intensity parameters were experimentally determined from emission spectrum of the [Eu(mpa)3(H2O)]complex and they were also calculated from crystallographic data. The complexes are coordination polymers, where the units of each complex are linked together by carboxylate groups leading to an unidimensional and parallel chains that by chemical interactions form a tridimensional framework. The emission spectrum profile of the [Eu(mpa)3(H2O)] complex is discussed based on point symmetry of the europium(III) ion, that explains the bands splitting observed in its emission spectrum. Photoluminescent analysis of the [Gd(mpa)3(H2O)] complex show no efficient ligand excitation but an intense charge transfer band. The excitation spectra of the [Eu(mpa)3(H2O)] and [Tb(mpa)3(H2O)] complexes do not show evidence of energy transfer from the ligand to the excited levels of these trivalent ions. Therefore the emission bands are originated only by direct f-f intraconfigurational excitation of the lantanide(III) ions.

Synthesis and photoluminescent properties of lanthanides acetoacetanilide complexes.

This work reports on the photoluminescent properties of three new lanthanide complexes with acetoacetanilide (aaa), a ß-diketonate ligand. The complexes have the general molecular formulae [RE(aaa)3(H2O)], they are soluble in organic solvents such as ethanol and chloroform and insoluble in water. The energy of the triplet state was determined at about 4,700 cm(-1) higher than the (5)D4 emitting level of the Tb(III) ion, leading to an absolute quantum yield of 22% for the [Tb(aaa)3(H2O)] complex. The photoluminescent properties were studied and the luminescence parameters of the [Eu(aaa)3(H2O)] complex were experimentally determined. The photostabilities of the complexes under continuous UV irradiation were measured and the data indicate low stability of the [Tb(aaa)3(H2O)] complex when the system is excited at the band attributed to energy transfer from the ligand to terbium(III) ion. However, its photostability is significantly improved under inert atmosphere.

Effects of 174 G/C polymorphism in the promoter region of the interleukin-6 gene on plasma IL-6 levels and muscle strength in elderly women

We investigated the effect of -174 G/C single-nucleotide polymorphism in the promoter region of the IL6 gene on plasma IL-6 levels and muscle strength, and the relationship between IL-6 levels and muscle strength in elderly women. The sample consisted of 199 elderly residents (73.0 ± 7.8 years old) from rest homes and the community in Belo Horizonte, MG, Brazil. -174 G/C polymorphism was determined by direct sequencing of the product by PCR, and plasma IL-6 concentrations were measured by ELISA. Muscle strength in the knee joint was evaluated using a Biodex System 3 Pro® isokinetic dynamometer. ANCOVA was used to determine the effect of polymorphism on IL-6 levels and muscle strength, and the Pearson correlation coefficient to assess the relationship between IL-6 levels and muscle strength. -174 G/C polymorphism was associated with the plasma IL-6 levels of elderly women (P < 0.01) since homozygotes for the G allele showed high IL-6 levels (GG 3.85 pg/mL, GC + CC 2.13 pg/mL). There was no association of polymorphism on muscle strength (P > 0.05). No association was found between IL-6 levels and knee extensor muscle (r = 0.087, P = 0.306) or flexor (r = -0.011, P = 0.894) strength. An interaction between -174 G/C polymorphism and housing conditions of the sample of elderly women was identified, with the effect of genotype on IL-6 levels being higher in the institutionalized elderly. These results support the evidence that -174 G/C polymorphism of the IL6 gene associates with individual variability of plasma IL-6 levels in elderly women.

Effects of 174 G/C polymorphism in the promoter region of the interleukin-6 gene on plasma IL-6 levels and muscle strength in elderly women.

We investigated the effect of -174 G/C single-nucleotide polymorphism in the promoter region of the IL6 gene on plasma IL-6 levels and muscle strength, and the relationship between IL-6 levels and muscle strength in elderly women. The sample consisted of 199 elderly residents (73.0 ± 7.8 years old) from rest homes and the community in Belo Horizonte, MG, Brazil. -174 G/C polymorphism was determined by direct sequencing of the product by PCR, and plasma IL-6 concentrations were measured by ELISA. Muscle strength in the knee joint was evaluated using a Biodex System 3 Pro® isokinetic dynamometer. ANCOVA was used to determine the effect of polymorphism on IL-6 levels and muscle strength, and the Pearson correlation coefficient to assess the relationship between IL-6 levels and muscle strength. -174 G/C polymorphism was associated with the plasma IL-6 levels of elderly women (P < 0.01) since homozygotes for the G allele showed high IL-6 levels (GG 3.85 pg/mL, GC + CC 2.13 pg/mL). There was no association of polymorphism on muscle strength (P > 0.05). No association was found between IL-6 levels and knee extensor muscle (r = 0.087, P = 0.306) or flexor (r = -0.011, P = 0.894) strength. An interaction between -174 G/C polymorphism and housing conditions of the sample of elderly women was identified, with the effect of genotype on IL-6 levels being higher in the institutionalized elderly. These results support the evidence that -174 G/C polymorphism of the IL6 gene associates with individual variability of plasma IL-6 levels in elderly women.

CompCytogen of two of the smallest Amazonian fishes: Fluviphylax simplex Costa, 1996 and Fluviphylax zonatus Costa, 1996 (Cyprinodontiformes, Poeciliidae).

The genus Fluviphylax Whitley, 1965is comprized of five valid species (Fluviphylax pygmaeus Myers et Carvalho, 1955, Fluviphylax zonatus, Fluviphylax simplex, Fluviphylax obscurus Costa, 1996,and Fluviphylax palikur Costa et Le Bail, 1999), which are endemic to the Amazon region. These fishes are the smallest known South American vertebrates and among the smallest know vertebrates on Earth. All species but the type Fluviphylax pygmaeus have been described in late 1990's, and much remains unknown about the biology, taxonomy and systematics of this group of fishes. The aims of the present study were to establish the diploid and haploid number of Fluviphylax zonatus and Fluviphylax simplex, and to find species-specific markers for the discrimination of taxa. The diploid number for both species was 48 chromosomes, with no sex chromosome heteromorphism. Fluviphylax zonatus exhibited the karyotypic formula 4m+8sm+22st+14a and FN=82, and Fluviphylax simplex exhibited 4m+16sm+18st+10a and FN=86. The determination of the total mean length of the chromosomes and their grouping into five size classes demonstrated different chromosome composition of the two species. This difference was further supported by the distribution of constitutive heterochromatin. The meiotic analysis revealed 24 bivalents in both species, but Fluviphylax zonatus exhibited chromosomes with late pairing of the telomeric portions in the pachytene. These data reveal that cytogenetic characterization is useful and important for the discrimination of these species. Our study further indicates that this method could be employed in the analysis of other species of small fishes that are difficult to distinguish using traditional morphological traits or are morphologically cryptic.

Dietary fat and apolipoprotein genotypes modulate plasma lipoprotein levels in Brazilian elderly women.

Studies show that genetic polymorphisms in apolipoproteins, which are in charge of lipid transport, predispose to atherogenic dyslipidemia. This study aimed to investigate the impact of apolipoprotein E, A5, and B genotypes and dietary intake on lipid profile in a sample of elderly women in Brazil. Two hundred and fifty-two women (60 years or older) living in the outskirts of the Brazilian Federal District underwent clinical and laboratory assessments to characterize glycemic and lipidemic variables, and also to exclude confounding factors (smoking, drinking, hormone replacement, cognitive impairment, physical activity). Three-day food records were used to determine usual dietary intake, whereas genotypic evaluations were in accordance to established methodologies. Genotype frequencies were consistent with the Hardy-Weinberg equilibrium. Prior to adjustment, individuals carrying the epsilon2 allele showed higher serum levels of triglycerides (P<0.05) and VLDL (P<0.005) compared to epsilon4 carriers, whereas LDL levels were considerably elevated in epsilon4 compared to epsilon2 carriers. In the presence of high intake of total fat or a low ratio of polyunsaturated to saturated fatty acid, epsilon4 carriers lost protection against hypertriglyceridemia. There was no association of the apolipoprotein A5 and B genotypes with lipidemic levels independently of the fat intake regimen. Results are suggestive of a dysbetalipoproteinemic-like phenotype in postmenopausal women, with remarkable gene-diet interaction.

Long-term outcome of renal transplantation at a university center in Rio de Janeiro between 1975 and 2000.

This study reports the 25-year experience of a single university center with respect to the impact of selected variables on long-term survival and half-life of 742 transplants. We calculated 1-, 5-, and 10-year Kaplan-Meier survival rates for grafts and patients, with separate analyses for HLA match and for each quinquennium. We also investigated the impact of cyclosporine administration and OKT3 induction. Global graft and patient survival rates were 85.2%, 61.0%, and 43.5% and 93.0%, 78.9%, and 66.6% for 1, 5, and 10 years, respectively. The half-life of 23.3 years for the 105 HLA-identical transplants was significantly better (P <.0001) than that for all other matches. Half-lives for 319 1-haplotype matched, 116 living-unrelated, and 153 cadaver grafts were 8.2, 5.7, and 5.6 years, respectively. No survival advantage was noted among these other matches. Introduction of cyclosporine in all non HLA-identical transplants and OKT3 induction for cadaver grafts conferred significant survival advantages compared with no use (P =.0002 and P =.009). There has been a continuous improvement in the long-term results in each quinquennium studied.

Is hepatitis C a risk factor to posttransplant diabetes mellitus after renal transplantation in patients using tacrolimus?

The objective of this study was to evaluate the association between previous hepatitis C virus (HCV) infection and the occurrence of posttransplant diabetes mellitus (PTDM) among patients undergoing kidney transplants using tacrolimus (FK). From August 1999 to January 2003, 66 patients (36.4 +/- 15.5 years) underwent kidney transplantation using an immunosuppressive regimen of tacrolimus, mycophenolate mofetil, or azathioprine and steroids. Thirty-four patients (52%) received kidneys from living donors and 32 (48%) from cadaveric donors. The diagnosis of diabetes mellitus was established after two consecutive ambulatory measurements of fasting glycemia > or = 126 mg/dL. Thirty-five percent of the patients (23/66) were HCV+ and 65% (43/66) HCV-. Of the 66 patients, 33% (22) developed PTDM, 19 (82%) from the HCV+ group and only 3 (7%) from the HCV- group. Among those who developed PDTM, the diagnosis was established in the first 2 posttransplant months in most cases (68.2%). The results showed a significant association between HCV and PTDM (P < or = .0001). In this group of patients HCV infection was strongly associated with the development of PTDM. Therefore, additional care is required regarding the immunosuppressive regimen among patients with chronic HCV infection.

Determinação do teor de umidade e presença de nitrito em amostras de charque / Determination of tenor of humidity and nitrite in dried meat samples

O nitrito de sódio é um aditivo que, ultimamente, vem preocupando a comunidade científica mundial devido a sua capacidade de combinar-se com aminas secundárias e terciárias, formando N-nitrosaminas, consideradas potentes carcinógenos, além de apresentarem efeitos teratogênicos e mutagênicos. Teve como objetivo determinar a concentração de nitrito e o teor de umidade de amostras de charque provenientes de diferentes estabelecimentos. Foram analisadas 24 amostras, provenientes de estabelecimentos comerciais do estado do Rio de Janeiro, classificados como A (grandes redes de supermercados) e B (pequenos comerciantes). O teor de umidade foi determinado pelo método da estufa a 105ÝC (Brasil, 1981) e, o nitrito, pelo método de Griess-llosvay (Brasil, 1981). Os resultados demonstraram que 16,66 por cento das amostras provenientes de estabelecimentos "A" apresentaram teor de nitrito em torno de 50ppm, 41,67 por cento acima deste valor e 41,67 por cento aproximadamente 10ppm. As amostras provenientes de estabelecimentos classificados como "B" apresentaram os resultados divididos em 33,33 por cento para quantidade de nitrito em torno de 50ppm, acima de 50ppm e 10 ppm. Em relação ao teor de umidade, observou-se que 100 por cento das amostras encontravam-se com teor de umidade acima de 45 por cento. A partir dos resultados obtidos, conclui-se que existe a necessidade de um maior controle no processamento do charque por parte das autoridades competentes, a fim de garantir um produto dentro das especificações estabelecidas pela legislação vigente. Entretanto, devido à aceitação de tal produto pelo mercado consumidor, sugere-se uma avaliação do processamento do charque, para uma possível reformulação de tais especificações, de acordo com a preferência do consumidor, desde que não tragam danos à saúde do mesmo.

[Evaluation of antiparasitic activity of allopurinol, against Trypanosoma cruzi, in experimental system using infected triatomines]. / Avaliação da atividade antiparasitária do alopurinol, referente ao Trypanosoma cruzi, em sistema experimental que utiliza triatomíneos infectados.

The antiparasitic activity of allopurinol, against Trypanosoma cruzi, was evaluated by a procedure using infected triatomines. This methodology indicated that the drug was unable to eliminate the protozoa in the digestive tract of the insects. Therefore, further knowledge to improve our understanding of allopurinol in the context of the etiologic treatment of infection by T. cruzi was not acquired. Despite this finding the drug continues to be used, even though its performance appears to be full of divergences and contradictions.