Morfologia e desenvolvimento ultraestrutural do sistema renal de embriões bovinos com idade gestacional entre 10 e 50 dias

O presente estudo teve como objetivo descrever o desenvolvimento dos sistemas renais de bovinos durante o período embrionário compreendido entre 10 e 50 dias. Embriões bovinos coletados em frigorífico foram fotografados e medidos utilizando-se o método Crow-Rump (CR) para estimar a idade gestacional. Os embriões destinados à miscroscopia óptica foram fixados em solução de Bouin para a avaliação do desenvolvimento do sistema renal, assim como suas estruturas. Alguns embriões também foram fixados em Glutaraldeído 2,5% e destinados à microscopia eletrônica de transmissão para o estudo ultraestrutural das células do sistema renal. Embriões entre o 14° e o 15° dia de desenvolvimento (E14-15) não apresentaram pronefro, mas apresentaram mesonefro, assim como indícios morfológicos que indicam sua atividade funcional. O mesonefro apresentou, no interior de suas células tubulares, inúmeras mitocôndrias e interdigitações, indicando uma alta atividade de transporte iônico. O metanefro, ou rim definitivo, iniciou seu desenvolvimento em E23-24. Os achados emonstram que a involução do mesonefro acontece simultaneamente com a diferenciação metanefrogênica. Em E45-46, já iniciando a fase fetal, o metanefro possuiu unidades filtradoras (néfrons), com seus respectivos glomérulos, túbulos contorcidos proximais e distais e alça de Henle. Nessa fase, o rim ainda não apresenta lobação externa.

Parâmetros genéticos para idade ao primeiro parto estimados por diferentes modelos para rebanhos da raça Nelore

Com objetivo de estimar parâmetros genéticos e estudar a utilização de diferentes efeitos em avaliações genéticas para idade ao primeiro parto (IPP) por diferentes modelos, foram utilizados registros de IPP de animais da raça Nelore, nascidos entre os anos de 1990 e 2005. Foram considerados os seguintes modelos (M): M1, incluindo o efeito fixo de GC1 (constituído pelos animais nascidos na mesma fazenda e ano), além da covariável, peso aos 365 dias de idade (efeito linear e quadrático), totalizando 24.263 registros de IPP; M2, considerando os efeitos fixos de GC1, ano e estação de parição, totalizando 59.792 registros de IPP e M3, incluindo os efeitos fixos de GC2 (agrupando os animais nascidos na mesma fazenda, ano e que conceberam no mesmo manejo reprodutivo), ano e estação de parição, totalizando 59.792 registros de IPP. As estimativas dos componentes de variância e herdabilidade e os valores genéticos (VG) foram obtidos pelo método da máxima verossimilhança restrita, com a inclusão da matriz de parentesco disponível. As diferenças esperadas na progênie (DEPs) foram obtidas dividindo os VG por dois. Após a obtenção desses resultados, foram realizadas correlações entre os VG e o ranqueamento das DEPs dos reprodutores para IPP, utilizando-se o procedimento PROC CORR (SAS, 2003). Ao se considerar o ano e a estação de parto nos modelos de análise (M2 e M3), esses produziram um maior R², indicando que tais modelos conseguiram explicar, em maior grau, as diferenças existentes entre os animais para IPP. As herdabilidades estimadas foram de baixa magnitude (0,14 e 0,15). As correlações entre os VG obtidas por diferentes modelos foram 0,73 (M1 x M2); 0,91 (M2 x M3) e 0,66 (M1 x M3).

Bovine gene polymorphisms related to fat deposition and meat tenderness

Leptin, thyroglobulin and diacylglycerol O-acyltransferase play important roles in fat metabolism. Fat deposition has an influence on meat quality and consumers' choice. The aim of this study was to determine allele and genotype frequencies of polymorphisms of the bovine genes, which encode leptin (LEP), thyroglobulin (TG) and diacylglycerol O-acyltransferase (DGAT1). A further objective was to establish the effects of these polymorphisms on meat characteristics. We genotyped 147 animals belonging to the Nelore (Bos indicus), Canchim (5/8 Bos taurus + 3/8 Bos indicus), Rubia Gallega X Nelore (1/2 Bos taurus + 1/2 Bos indicus), Brangus Three-way cross (9/16 Bos taurus + 7/16 Bos indicus) and Braunvieh Three-way cross (3/4 Bos taurus + 1/4 Bos indicus) breeds. Backfat thickness, total lipids, marbling score, ribeye area and shear force were fitted, using the General Linear Model (GLM) procedure of the SAS software. The least square means of genotypes and genetic groups were compared using Tukey's test. Allele frequencies vary among the genetic groups, depending on Bos indicus versus Bos taurus influence. The LEP polymorphism segregates in pure Bos indicus Nelore animals, which is a new finding. The T allele of TG is fixed in Nelore, and DGAT1 segregates in all groups, but the frequency of allele A is lower in Nelore animals. The results showed no association between the genotypes and traits studied, but a genetic group effect on these traits was found. So, the genetic background remains relevant for fat deposition and meat tenderness, but the gene markers developed for Bos taurus may be insufficient for Bos indicus.

Damage assessment of the equine sperm membranes by fluorimetric technique

To validate a practical technique of simultaneous evaluation of the plasma, acrosomal and mitochondrial membranes in equine spermatozoa three fluorescent probes (PI, FITC-PSA and MITO) were associated. Four ejaculates from three stallions (n=12) were diluted in TALP medium and split into 2 aliquots, 1 aliquot was flash frozen in liquid nitrogen to induce damage in cellular membranes. Three treatments were prepared with the following fixed ratios of fresh semen: flash frozen semen: 100:0 (T100), 50:50 (T50), and 0:100 (T0). A 150-µL aliquot of diluted semen of each treatment was added of 2 µL of PI, 2 µL of MITO and 80 µL of FITC-PSA; incubated at 38.5ºC/8 min, and sperm cells were evaluated by epifluorescent microscopy. Based in regression analysis, this could be an efficient and practical technique to assess damage in equine spermatozoa, as it was able to determine the sperm percentage more representative of the potential to fertilize the oocyte.

Associação da medroxiprogesterona ao protocolo Ovsynch para inseminação artificial em tempo fixo de búfalas cíclicas (Bubalus bubalis) criadas na Amazônia Oriental

O presente estudo visou avaliar os efeitos da associação da medroxiprogesterona (análogo sintético da progesterona) ao protocolo Ovsynch sobre o crescimento folicular, a ovulação e a taxa de concepção de búfalas criadas na Amazônia Oriental (Tracuateua-PA). Vinte e sete fêmeas adultas (G1 n=14 e G2 n=13), cíclicas, sem bezerro ao pé e com ECC 3,5 foram submetidas a Ovsynch. Os animais do G2 receberam 60 mg de medroxiprogesterona entre D0 e D7 (D0=início do tratamento). A ultra-sonografia ovariana foi realizada nos D 0, 7, 9 e 10. O contingente de folículos pequenos diferiu no D7 (G1: 4,57±0,60 versus G2: 6,54±0,67; P=0,05). Tempo e tratamento influenciaram o diâmetro folicular no D7. O crescimento do folículo dominante entre D7 e D9 foi maior nos animais tratados (G1: 2,05±0,49 mm/dia versus 3,48±0,41 mm/dia; P<0,05). Mais animais do G1 ovularam precocemente (35,71% versus 30,77%), porém isso não afetou as taxas de concepção (G1: 50,00% e G2: 30,77%; P>0,05). Os achados sugerem que a medroxiprogesterona (1) aumenta recrutamento folicular e retarda o crescimento dos folículos com diâmetro maior que 5,0 mm entre D0 e D7; (2) sua retirada incrementa em 1,7 vezes o crescimento folicular do D7 ao D9; (3) pode contribuir para a ovulação de folículos maiores e, em tese, para maior formação de tecido luteínico; (4) não promove ovulação precoce após o Ovsynch; (5) não eleva as taxas de concepção após sincronização de fêmeas cíclicas e com bom escore corporal, devendo ser avaliada para uso em fêmeas acíclicas ou com ECC mais baixo.

Modelo de suprimento sanguíneo do intestino grosso do tamanduá bandeira (Myrmecophaga tridactyla)

Em seis animais adultos Myrmecophaga tridactyla estudou-se o modelo de suprimento sanguíneo do intestino grosso, que é dependente das artérias mesentérica cranial (AMCr) e caudal (AMC). Os espécimes coletados conforme as normas do IBAMA (Instituto Brasileiro do Meio Ambiente e dos Recursos Naturais Renováveis) foram perfundidos com água (40 C), injetados com látex corado, fixados em formol e conservados em solução alcoólica (50%). O mesocólon está disposto na linha sagital mediana e fixa o intestino grosso à parede dorsal do abdome. Derivaram da AMC: a artéria retal cranial, sete a 14 artérias cólicas e uma ou duas artérias ileocólicas, que apresentaram anastomoses de irrigação com a AMCr. A AMC finaliza-se na borda mesocólica das alças intestinais, emitindo ramos cólicos retos a partir das arcadas justacólicas, que penetram na intimidade da musculatura longitudinal. Ao longo do trajeto da AMC foram observadas ilhas arteriais, e a região ileocólica apresentou maior densidade vascular.

Creatine supplementation spares muscle glycogen during high intensity intermittent exercise in rats

Background: The effects of creatine (CR) supplementation on glycogen content are still debatable. Thus, due to the current lack of clarity, we investigated the effects of CR supplementation on muscle glycogen content after high intensity intermittent exercise in rats. Methods: First, the animals were submitted to a high intensity intermittent maximal swimming exercise protocol to ensure that CR-supplementation was able to delay fatigue ( experiment 1). Then, the CR-mediated glycogen sparing effect was examined using a high intensity intermittent sub-maximal exercise test ( fixed number of bouts; six bouts of 30-second duration interspersed by two-minute rest interval) ( experiment 2). For both experiments, male Wistar rats were given either CR supplementation or placebo (Pl) for 5 days. Results: As expected, CR-supplemented animals were able to exercise for a significant higher number of bouts than Pl. Experiment 2 revealed a higher gastrocnemius glycogen content for the CR vs. the Pl group (33.59%). Additionally, CR animals presented lower blood lactate concentrations throughout the intermittent exercise bouts compared to Pl. No difference was found between groups in soleus glycogen content. Conclusion: The major finding of this study is that CR supplementation was able to spare muscle glycogen during a high intensity intermittent exercise in rats.

Nutritive value and physical characteristics of Xaraes palisadegrass as affected by grazing strategy

The aim of this study was to ascertain whether the defoliation frequency based on a fixed rest period would generate variable sward structural and physiological conditions at each subsequent grazing event. The relative importance of the physiological age was established in comparison with the chronological age in the determination of the forage nutritive value of Xaraes palisadegrass [Brachiaria brizantha (Hochst ex A. RICH.) STAPF. cv. Xaraes]. Two grazing frequencies were defined by light interception (LI) at initiation of grazing (95% LI - ""target grazing"" [TG] or 100% LI - ""delayed grazing"" [DG]) and one based on chronological time, grazing every 28 days (28-d). Forage produced under the TG schedule was mostly leaves (93%) with a higher concentration of crude protein (CP; 138 g/kg in the whole forage), a lower concentrations of neutral detergent fibre (NDF) in the stems (740 g/kg), and higher in vitro dry matter digestibility (IVDMD) of the leaves (690 g/kg), compared to the other treatments. Lower grazing frequency strategies (DG and 28-d) resulted in forage with higher proportions of stems (10 and 9%, respectively). Strategies based on light interception did not produce pre-graze forage with a uniform nutritive value, as the indicators varied across grazing cycles. The treatment based on fixed days of rest did not result in uniformity.

Non-additive genetic effects on weights and performance of a Brazilian Bos taurus x Bos indicus beef composite

The aim of the present study was to evaluate the heterosis effects on weaning weight at 205 days (WW, N = 146,464), yearling weight at 390 days (YW, N = 69,315) and weight gain from weaning to yearling (WG, N = 59,307) in composite beef cattle. The fixed models were: RM, which included contemporary groups, class of age of dam, outcrossing percentages for direct and maternal effects, and additive direct and maternal ( AM) breed effects; R, RM model, minus AM breed effects, and H, RM model, minus additive breed effects. The estimates for W205 were in general positive (P < 0.01). The R and H models resulted in similar estimates, but they were very different from the ones estimated by the RM model. For W390, the R and H models resulted in general positive estimates (P < 0.05). For WG, the RM model resulted in general significant heterosis effects (P < 0.05). It can be concluded that the RM model seems to supply estimates of better quality (P < 0.01).

Histometry of the sublingual gland in male and female mice (Mus musculus) infected with the RAL strain of the Chagas parasite, Trypanosoma cruzi

Trypanosoma cruzi. The aim of this work was to analyze histologically and histometrically the sublingual gland of mice infected with the RAL strain of T cruzi, according to the sex. Swiss mice (Mus musculus) were inoculated with 2 x 10(4) blood trypomastigotes of the RAL strain of T cruzi. In the peak of the parasitemia (12th day) the mice were sacrificed, and the sublingual glands were fixed in ALFAC. HE-stained histological sections were evaluated histometrically. The parasitemia was higher in females. Histopatologically, acini of the infected animals were smaller, with scanty production of secretion, and smaller striated ducts. The nuclei of the demilunes were smaller and showed amastigote nests in the cytoplasm. Karyometrically, nuclei of the acini, demilunes and striated ducts were smaller in the infected mice. Stereologically, it was observed that relative volumes of acini and ducts were smaller and, inversely, relative volumen were greater for the conjunctive tissue in the infected males. The surface densities of acini and ducts were bigger and the diameter and thickness of the wall were smaller in this group. On the other hand, relative volume of acini was smaller and those of the ducts and conjunctive tissue were bigger in the infected females. The diameter and thickness of the wall of acini were smaller, and those of the striated ducts were bigger in this group. The RAL strain of T cruzi caused general atrophy in the sublingual gland, with numerous nests of parasites in the glandular parenchyma.

Analysis of breast cancer by small angle X-ray scattering (SAXS)

Small angle X-ray scattering (SAXS) images of normal breast tissue and benign and malignant breast tumour tissues, fixed in formalin, were measured at the momentum transfer range of 0.063 nm(-1) <= q (=4 pi sin(theta/2)/lambda) <= 2.720 nm(-1). Four intrinsic parameters were extracted from the scattering profiles (1D SAXS image reduced) and, from the combination of these parameters, another three parameters were also created. All parameters, intrinsic and derived, were subject to discriminant analysis, and it was verified that parameters such as the area of diffuse scatter at the momentum transfer range 0.50 <= q <= 0.56 nm(-1), the ratio between areas of fifth-order axial and third-order lateral peaks and third-order axial spacing provide the most significant information for diagnosis (p < 0.001). Thus, in this work it was verified that by combining these three parameters it was possible to classify human breast tissues as normal, benign lesion or malignant lesion with a sensitivity of 83% and a specificity of 100%.

Forward Neutral-Pion Transverse Single-Spin Asymmetries in p plus p Collisions at s=200 GeV

We report precision measurements of the Feynman x (x(F)) dependence, and first measurements of the transverse momentum (p(T)) dependence, of transverse single-spin asymmetries for the production of pi(0) mesons from polarized proton collisions at s=200 GeV. The x(F) dependence of the results is in fair agreement with perturbative QCD model calculations that identify orbital motion of quarks and gluons within the proton as the origin of the spin effects. Results for the p(T) dependence at fixed x(F) are not consistent with these same perturbative QCD-based calculations.

249 TP53 mutation has high prevalence and is correlated with larger and poorly differentiated HCC in Brazilian patients

Background: Ser-249 TP53 mutation (249(Ser)) is a molecular evidence for aflatoxin-related carcinogenesis in Hepatocellular Carcinoma (HCC) and it is frequent in some African and Asian regions, but it is unusual in Western countries. HBV has been claimed to add a synergic effect on genesis of this particular mutation with aflatoxin. The aim of this study was to investigate the frequency of 249Ser mutation in HCC from patients in Brazil. Methods: We studied 74 HCC formalin fixed paraffin blocks samples of patients whom underwent surgical resection in Brazil. 249Ser mutation was analyzed by RFLP and DNA sequencing. HBV DNA presence was determined by Real-Time PCR. Results: 249Ser mutation was found in 21/74 (28%) samples while HBV DNA was detected in 13/74 (16%). 249Ser mutation was detected in 21/74 samples by RFLP assay, of which 14 were confirmed by 249Ser mutant-specific PCR, and 12 by nucleic acid sequencing. All HCC cases with p53-249ser mutation displayed also wild-type p53 sequences. Poorly differentiated HCC was more likely to have 249Ser mutation (OR = 2.415, 95% CI = 1.001 - 5.824, p = 0.05). The mean size of 249Ser HCC tumor was 9.4 cm versus 5.5 cm on wild type HCC (p = 0.012). HBV DNA detection was not related to 249Ser mutation. Conclusion: Our results indicate that 249Ser mutation is a HCC important factor of carcinogenesis in Brazil and it is associated to large and poorly differentiated tumors.

Development and mechanical testing of a short intramedullary nail for fixation of femoral rotational osteotomy in cerebral palsy patients

Background: Rotational osteotomy is frequently indicated to correct excessive femoral anteversion in cerebral palsy patients. Angled blade plate is the standard fixation device used when performed in the proximal femur, but extensile exposure is required for plate accommodation. The authors developed a short locked intramedullary nail to be applied percutaneously in the fixation of femoral rotational osteotomies in children with cerebral palsy and evaluated its mechanical properties. Methods: The study was divided into three stages. In the first part, a prototype was designed and made based on radiographic measurements of the femoral medullary canal of ten-year-old patients. In the second, synthetic femoral models based on rapid-prototyping of 3D reconstructed images of patients with cerebral palsy were obtained and were employed to adjust the nail prototype to the morphological changes observed in this disease. In the third, rotational osteotomies were simulated using synthetic femoral models stabilized by the nail and by the AO-ASIF fixed-angle blade plate. Mechanical testing was done comparing both devices in bending-compression and torsion. Results: The authors observed proper adaptation of the nail to normal and morphologically altered femoral models, and during the simulated osteotomies. Stiffness in bending-compression was significantly higher in the group fixed by the plate (388.97 +/- 57.25 N/mm) than in that fixed by the nail (268.26 +/- 38.51 N/mm) as torsional relative stiffness was significantly higher in the group fixed by the plate (1.07 +/- 0.36 Nm/degrees) than by the nail (0.35 +/- 0.13 Nm/degrees). Conclusions: Although the device presented adequate design and dimension to fit into the pediatric femur, mechanical tests indicated that the nail was less stable than the blade plate in bending-compression and torsion. This may be a beneficial property, and it can be attributed to the more flexible fixation found in intramedullary devices.

Suppression subtractive hybridization analysis reveals expression of conserved and novel genes in male accessory glands of the ant Leptothorax gredleri

Background: During mating, insect males eject accessory gland proteins (Acps) into the female genital tract. These substances are known to affect female post-mating behavior and physiology. In addition, they may harm the female, e. g., in reducing its lifespan. This is interpreted as a consequence of sexual antagonistic co-evolution. Whereas sexual conflict abounds in non-social species, the peculiar life history of social insects (ants, bees, wasps) with lifelong pair-bonding and no re-mating aligns the reproductive interests of the sexes. Harming the female during mating would negatively affect male fitness and sexual antagonism is therefore not expected. Indeed, mating appears to increase female longevity in at least one ant species. Acps are presumed to play a role in this phenomenon, but the underlying mechanisms are unknown. In this study, we investigated genes, which are preferentially expressed in male accessory glands of the ant Leptothorax gredleri, to determine which proteins might be transferred in the seminal fluid. Results: By a suppression subtractive hybridization protocol we obtained 20 unique sequences (USs). Twelve had mutual best matches with genes predicted for Apis mellifera and Nasonia vitripennis. Functional information (Gene Ontology) was available only for seven of these, including intracellular signaling, energy-dependent transport and metabolic enzyme activities. The remaining eight USs did not match sequences from other species. Six genes were further analyzed by quantitative RT-PCR in three life cycle stages of male ants. A gene with carboxy-lyase activity and one of unpredicted function were significantly overexpressed in accessory glands of sexually mature males. Conclusions: Our study is the first one to investigate differential gene expression in ants in a context related to mating. Our findings indicate that male accessory glands of L. gredleri express a series of genes that are unique to this species, possibly representing novel genes, in addition to conserved ones for which functions can be predicted. Identifying differentially expressed genes might help to better understand molecular mechanisms involved in reproductive processes in eusocial Hymenoptera. While the novel genes could account for rapidly evolving ones driven by intra-sexual conflict between males, conserved genes imply that rather beneficial traits might get fixed by a process described as inter-sexual cooperation between males and females.