Effects of antifreeze protein from Lolium perenne L. (LpAFP) in the vitrification of in vitro-produced bovine embryos.

In the present study, the cryoprotective effects of Lolium perenne antifreeze protein (LpAFP) on the vitrification of bovine embryos were evaluated. In vitro-produced blastocysts were divided into two groups: the control group (CG) without the addition of LpAFP and the treatment group (TG) with the addition of 500 ng/ml of LpAFP in the equilibrium and vitrification solution. Vitrification was carried out by transferring the blastocysts to the equilibrium solution [7.5% ethylene glycol (EG) and 7.5% dimethyl sulfoxide (DMSO)] for 2 min and then to the vitrification solution (15% EG, 15% DMSO and 0.5M sucrose). The blastocysts were deposited on a cryotop device and submerged in liquid nitrogen. Warming was carried out in three steps in solutions with different sucrose concentrations (1.0, 0.5, and 0.0 M, respectively). Embryos were evaluated for re-expansion/hatching, the total cell count, and ultrastructural analysis. There was no significant difference in the re-expansion rate 24 h after warming; however, there was variation (P < 0.05) in the hatching rate in the TG and the total number of cells 24 h after warming was higher in the TG (114.87 ± 7.24) when compared with the CG (91.81 ± 4.94). The ultrastructural analysis showed changes in organelles related to the vitrification process but, in the TG, there was less damage to mitochondria and rough endoplasmic reticulum compared with the CG. In conclusion, the addition of 500 ng/ml of LpAFP during the vitrification of in vitro-produced bovine embryos improved the hatching rate and total cell number of blastocysts after warming and mitigated intracellular damage.

PI3K inhibition by BKM120 results in anti-proliferative effects on corticotroph tumor cells.

PURPOSE: Cushing's disease is associated with significant morbidity; thus, additional tumor-directed drugs with the potential to exert antineoplastic effects on corticotroph adenoma cells are desired. The phosphoinositide-3-kinase (PI3K)/protein kinase B (AKT) pathway, which plays regulatory role in cell survival and proliferation, is activated in pituitary adenomas. The present study evaluated the effects of BKM120 (Buparlisib), an oral PI3K inhibitor, on cell viability, apoptosis, cell cycle phase distribution, and ACTH production in mouse corticotroph tumor cells. METHODS: AtT-20/D16v-F2 mouse pituitary corticotroph tumor cells were treated with increasing concentrations of BKM120 or vehicle. Cell viability was measured using an MTS-based assay. Apoptosis was evaluated by Annexin V staining. Cell cycle analysis was performed by propidium iodide DNA staining and flow cytometry. Gene expression of cell cycle regulators (Cdkn1b, Ccnd1, Ccne1, Cdk2, Cdk4, Myc, and Rb1) was assessed by qPCR. Protein expression of p27, total and phosphorylated Akt was assessed by Western blot. ACTH levels were measured in the culture supernatants by chemiluminescent immunometric assay. RESULTS: Treatment with BKM120 decreased AtT-20/D16v-F2 cell viability, induced a G0/G1 cell cycle arrest, reduced the phosphorylation of Akt at Serine 473, and increased p27 expression. Furthermore, BKM120 treatment diminished ACTH levels in the cell culture supernatants. CONCLUSION: In vitro inhibition of PI3K/AKT pathway by BKM120 resulted in anti-proliferative effects on corticotroph tumor cells, decreasing cell viability and ACTH production. These encouraging findings shape the path for further experiments with the inhibition of PI3K/AKT pathway in Cushing's disease.

Bioecological aspects of triatomines and marsupials as wild Trypanosoma cruzi reservoirs in urban, peri-urban and rural areas in the Western Brazilian Amazon.

In the Amazon region, Trypanosoma cruzi transmission cycles involve a great diversity of Triatominae vectors and mammal reservoirs. Some Rhodnius spp. mainly inhabit palm trees that act as microhabitats for hosts and vectors. The current study aimed to describe aspects of the bio-ecology of the vectors and reservoirs of T. cruzi in relation to human populations resident near areas with large quantities of palm trees, in rural, peri-urban and urban collection environments, located in the Western Brazilian Amazon. Rhodnius pictipes and Didelphis marsupialis were respectively the most predominant vector and reservoir, with rates of 71% for R. pictipes and 96.5% for D. marsupialis. The vast majority of T. cruzi isolates clustered with TcI. The most prevalent haplotype was TcI COII1 (69.7%). Mauritia flexuosa and Attalea phalerata were the main ecological indicators of infestation by triatomines. Birds were the most common food source (27,71%). T. cruzi isolated from R. robustus has the haplotype HUM-13, previously detected in a chronic Chagas patient living in the same area. Our results demonstrate the relevance of this study, with the occurrence of elevated infection rates in animals, and suggest the importance of the Amazon zones where there is a risk of infection in humans.

Zinc, manganese, and copper amino acid complexes improve performance and bone characteristics of layer-type chicks under thermoneutral and cold stress conditions.

Two experiments were designed to evaluate the effect of mineral-amino acid complexes (AACM) as a partial replacement of inorganic mineral (IM) in layer-type chicks' diets. Both studies had the same dietary treatments, where in experiment 1 (Exp. 1) was conducted under thermoneutral conditions from 0 to 35 D and chicks in experiment 2 (Exp. 2) were exposed to cold stress conditions at nighttime during the first 15 D and to thermoneutral condition from 16 to 35 D. For each trial, 1,200 one-day-old Lohmann Brown chicks were used, with 20 cage replicates with 30 chicks per cage. Treatments consisted of the control diet (IM; with 70, 70, and 8 mg/kg of zinc [Zn], manganese [Mn], and copper [Cu], respectively) and the treatment diet (AACM, with 40, 40, and 2.75 mg/kg of Zn, Mn, and Cu, respectively, from IM sources, along with 30, 30, and 5.25 mg/kg of Zn, Mn, and Cu, respectively). Data were submitted to analysis of variance, and means were compared using the t-test (P < 0.05). In Exp. 1, there were no significant differences between treatments on chick performance. However, AACM-fed chicks had higher thymus (P = 0.03) and cecum weight (P < 0.01), superior micromineral deposition in the tibias (P < 0.01), and reduced phosphorus excretion (P = 0.03). In Exp. 2, chicks fed with AACM had higher body weight gain (P = 0.04), better average daily feed intake (P = 0.03), lower phosphorus excretion (P = 0.02), and higher liver and pancreas weight (P < 0.01) in the last week of the study. In conclusion, chicks fed with AACM under thermoneutral conditions had higher bone mineralization and reduced excretion of phosphorus, and in adverse conditions, AACM improves performance and liver and pancreas weight, also reducing phosphorus excretion.

True absolute determination of photoluminescence quantum yields by coupling multiwavelength thermal lens and photoluminescence spectroscopy.

Photoluminescence quantum yields denote a critical variable to characterise a fluorophore and its potential performance. Their determination, by means of methodologies employing reference standard materials, inevitably leads to large uncertainties. In response to this, herein we report for the first time an innovative and elegant methodology, whereby the use of neat solvent/reference material required by thermal lens approaches is eliminated by coupling it to photoluminescence spectroscopy, allowing for the discrimination between materials with similar photoluminescence quantum yields. To achieve this, both radiative and non-radiative transitions are simultaneously measured using a photoluminescence spectrometer coupled to a multiwavelength thermal lens spectroscopy setup in a mode-mismatched dual-beam configuration, respectively. The absorption factor independent ratio of the thermal lens and photoluminescence signals can then be used to determine the fluorescence quantum yield both accurately and precisely. We validated our reported method using rhodamine 6G and further applied it to three novel structurally related diketopyrrolopyrrole based materials, which, in contrast to results obtained by other methods, unveiled significant differences in their photoluminescence quantum yields.

Laser cooling of Yb3+:KYW.

We report the first observation of laser cooling in Yb3+:KYW and validate the results by comparison with experiments in the well-studied material Yb3+:YAG. Radiation from a single-mode Ti:Al2O3 laser was used to achieve cooling of 1.5 K/W in 1% Yb:KYW at 1025 nm, comparing well with the reference material 3% Yb:YAG which cooled by 3.5 K/W at 1030 nm under open lab conditions. Experimental results for KYW crystals mounted on aerogels and doped with 1-20% Yb were in excellent agreement with the theoretical dependence of cooling power on the Yb absorption spectrum. Elimination of thermal conduction through the sample support structure was found to permit the attainment of lower temperatures and to simplify modeling of radiation balance conditions in self-cooled lasers with longitudinal thermal gradients. Contrary to the notion that more coolant ions yield higher cooling power, concentrations of Yb over 1% caused re-absorption of luminescence in KYW crystals, leading to a progressive red shift in the optimal cooling wavelength and the prevention of laser cooling altogether in a 20% sample at room temperature. The prospect of attaining radiation-balanced lasing in commercially-available tungstate crystals is evaluated.

The molecular motor Myosin Va interacts with the cilia-centrosomal protein RPGRIP1L.

Myosin Va (MyoVa) is an actin-based molecular motor abundantly found at the centrosome. However, the role of MyoVa at this organelle has been elusive due to the lack of evidence on interacting partners or functional data. Herein, we combined yeast two-hybrid screen, biochemical studies and cellular assays to demonstrate that MyoVa interacts with RPGRIP1L, a cilia-centrosomal protein that controls ciliary signaling and positioning. MyoVa binds to the C2 domains of RPGRIP1L via residues located near or in the Rab11a-binding site, a conserved site in the globular tail domain (GTD) from class V myosins. According to proximity ligation assays, MyoVa and RPGRIP1L can interact near the cilium base in ciliated RPE cells. Furthermore, we showed that RPE cells expressing dominant-negative constructs of MyoVa are mostly unciliated, providing the first experimental evidence about a possible link between this molecular motor and cilia-related processes.

The influence of canine brucellosis on morphofunctional features of epididymal spermatozoa: case report / Influência da brucelose canina nos aspectos morfofuncionais de espermatozoides epididimários: relato de caso

The present work reports a clinical case of a mongrel dog, with serological diagnosis of brucellosis, from which epididymal sperm analysis was performed. Sperm samples were collected from different segments of the epididymis (tail, corpus, and caput). Sperm samples were evaluated for computer-assisted motility analysis (CASA), spermatic morphology, mitochondrial activity and sperm plasmatic membrane and acrosomal integrity. Changes in sperm movement patterns were found (progressive motility, percentage of rapid sperm, percentage of rapid velocity, average pathway, curvilinear velocity, velocity straight line, amplitude of lateral head displacement, straightness and linearity), increase of total morphological defects (51%) and absence of sperm mitochondrial activity (20%) were verified, especially for cauda epididymides. We highlight that such changes can contribute to clinical diagnosis of Brucellosis in dogs and to the use of epididymal sperm in reproductive biotechnologies.(AU)

Relata-se o caso de um cão mestiço, com diagnóstico sorológico para brucelose canina, a partir do qual foram realizadas análises do sêmen epididimário. As amostras espermáticas foram coletadas dos diferentes segmentos epididimários (cabeça, corpo e cauda). Foram realizadas as avaliações de motilidade computadorizada do sêmen (CASA), morfologia espermática, atividade mitocondrial, integridade das membranas plasmática e acrossomal. Houve alteração no padrão de movimentação espermática (motilidade progressiva, espermatozoides rápidos, velocidade média da trajetória, velocidade curvilínea, velocidade linear progressiva, amplitude de deslocamento lateral da cabeça, retilinearidade e linearidade), aumento do total de defeitos morfológicos (51%) e da ausência de atividade mitocondrial espermática (20%) dos espermatozoides, especialmente da cauda do epidídimo. Ressalta-se que tais achados podem contribuir para o diagnóstico clínico da brucelose canina e para a utilização do sêmen epididimário em biotecnologias da reprodução.(AU)

Energia metabolizável da levedura de cana-de-açúcar submetida a diferentes temperaturas e tempos de secagem para frangos de corte e poedeiras / Metabolizable energy of sugar cane yeast submitted to different temperatures and drying times for broilers and layers hens

Objetivou-se determinar a temperatura e o tempo de secagem por rolos rotativos, aos quais a, levedura de cana-de-açúcar é submetida que permitam seu melhor aproveitamento energético por galinhas poedeiras e frangos de corte. Para isso foram realizados três ensaios de metabolismo para determinar os valores de energia metabolizável aparente (EMA), aparente corrigida para nitrogênio (EMAn) e os coeficientes de metabolizabilidade aparente da matéria seca (CMMS) e da energia bruta (CMEB). O primeiro ensaio foi conduzido com galinhas poedeiras (E1), o segundo com frangos de corte (E2) em crescimento e o terceiro com frangos de corte em diferentes idades (E3)...

This study aimed to determine the temperature and drying time through rotative rolls, that sugar cane yeast is subjected to in order to allow best energy utilization by laying hens and broilers. Three metabolism trials were conducted to determine the values of apparent metabolizable energy (AME) and apparent corrected for nitrogen balance (AMEn), coefficient of apparent metabolizable dry matter (CAMDM) and gross energy (CAMGE). The first experiment was conducted with laying hens (E1), the second with broilers (E2) in growth and the third with broilers at different ages (E3)...

Species of Anastrepha (Diptera: Tephritidae) captured in a guava orchard (Psidium guajava L., Myrtaceae) in Boa Vista, Roraima, Brazil / Espécies de Anastrepha (Diptera: Tephritidae) capturadas em pomar de goiabeira (Psidium guajava L., Myrtaceae) em Boa Vista, Roraima, Brasil

The guava fruit (Psidium guajava) is among the most strongly affected by fruit flies in Brazil. In the Brazilian Amazon, 11 species of Anastrepha have been reported in guava orchards to date. This work aimed to identify the species of Anastrepha present in a guava orchard in the municipality of Boa Vista, determine the species infesting the fruits, and identify any parasitoids present. Two McPhail traps with food bait were installed and weekly collections were made between January and December 2008. Fruits were also collected systematically during this period, with a view to determining the association between host plant and tephritid species. Nine species of Anastrepha were identified, in addition to one specimen belonging to a probable new species. Anastrepha striata Schiner, Anastrepha sororcula Zucchi, Anastrepha obliqua (Macquart), and Anastrepha fraterculus (Wiedemann) were the dominant species in the orchard, accounting for 84.8% of all captured individuals. All females collected directly from fruits were A. striata. Doryctobracon areolatus (Szépligeti) was the only parasitoid species obtained. In this work, Anastrepha ethalea (Walker) is reported for the first time in the state of Roraima.

A goiaba (Psidium guajava) é uma das frutas mais afetadas pelas moscas-das-frutas no Brasil. Na Amazônia brasileira 11 espécies de Anastrepha já foram relatadas nesta cultura. O presente trabalho teve como objetivos identificar as espécies de Anastrepha presentes em um pomar de goiabeira no município de Boa Vista, verificar quais espécies infestam os frutos e identificar os parasitoides presentes. Foram instaladas duas armadilhas tipo McPhail, contendo atrativo alimentar, sendo as coletas realizadas semanalmente, de janeiro a dezembro de 2008. Para determinar a associação entre a planta hospedeira e as espécies de tefritídeos foram realizadas coletas sistemáticas de frutos no mesmo período. Nove espécies de Anastrepha foram identificadas, além dessas foi obtido um exemplar pertencente a uma provável espécie nova. Anastrepha striata Schiner, Anastrepha sororcula Zucchi, Anastrepha obliqua (Macquart) e Anastrepha fraterculus (Wiedemann) foram as espécies dominantes no pomar e representaram 84,8% dos indivíduos capturados. Todas as fêmeas coletadas diretamente dos frutos pertenciam à espécie A. striata. Doryctobracon areolatus (Szépligeti) foi a única espécie de parasitoide obtida. Neste trabalho, Anastrepha ethalea (Walker) é assinalada pela primeira vez no estado de Roraima.

Species of Anastrepha (Diptera: Tephritidae) captured in a guava orchard (Psidium guajava L., Myrtaceae) in Boa Vista, Roraima, Brazil.

The guava fruit (Psidium guajava) is among the most strongly affected by fruit flies in Brazil. In the Brazilian Amazon, 11 species of Anastrepha have been reported in guava orchards to date. This work aimed to identify the species of Anastrepha present in a guava orchard in the municipality of Boa Vista, determine the species infesting the fruits, and identify any parasitoids present. Two McPhail traps with food bait were installed and weekly collections were made between January and December 2008. Fruits were also collected systematically during this period, with a view to determining the association between host plant and tephritid species. Nine species of Anastrepha were identified, in addition to one specimen belonging to a probable new species. Anastrepha striata Schiner, Anastrepha sororcula Zucchi, Anastrepha obliqua (Macquart), and Anastrepha fraterculus (Wiedemann) were the dominant species in the orchard, accounting for 84.8% of all captured individuals. All females collected directly from fruits were A. striata. Doryctobracon areolatus (Szépligeti) was the only parasitoid species obtained. In this work, Anastrepha ethalea (Walker) is reported for the first time in the state of Roraima.

Cytogenetic damage in the buccal epithelium of Brazilian aviators occupationally exposed to agrochemicals.

The frequency of micronuclei in both buccal cells and peripheral blood lymphocytes is extensively used as a biomarker of chromosomal damage and genome stability in human populations. We examined whether prolonged exposure to complex mixtures of pesticides leads to an increase in cytogenetic damage. The exposed group comprised 50 agricultural aviators, mainly from Central and Southeast regions of Brazil, who had inhaled agrochemicals for more than 10 years without personal protection equipment; the control group consisted of 17 men from the same regions, without indication of exposure to pesticides, There were three times higher frequencies of micronuclei (P < 0.05) and 2.5 times higher frequencies of binucleated cells in the aviators when compared to controls. However, cytotoxic alterations such as broken eggs and karyorrhexis did not present statistically significant differences between the exposed and control groups. Therefore, diverse agrochemicals used to combat pests in agriculture possess genotoxic effects in the oral mucosa of the agricultural pilots, as showed in this study.

Substituição do óleo de peixe por óleo de soja em dietas para beijupirá (Rachycentron canadum) / Replacement of fish oil by soybean oil in diets for cobia (Rachycentron canadum)

A substituição do óleo de peixe em dietas para beijupirá (Rachycentron canadum) foi avaliada usando-se cinco proporções de óleo de peixe e soja. Duzentos juvenis (±12g) foram distribuídos aleatoriamente em 20 tanques e alimentados até a saciedade aparente durante 42 dias. À medida que se aumentou o teor de óleo de peixe nas dietas, houve aumento proporcional no desempenho. Os juvenis alimentados com dietas que continham 75 e 100 por cento de óleo de peixe apresentaram ganhos de peso mais altos que os das dietas com 0 e 25 por cento de óleo de peixe, enquanto os alimentados com a dieta com 50 por cento de óleo de peixe apresentaram crescimento intermediário. Não foram observadas diferenças nas concentrações de proteína e lipídio na carcaça dos peixes, mas os teores de umidade e cinzas diferiram entre os tratamentos. A inclusão de teores relativamente altos de ingredientes de origem vegetal diminuiu o consumo alimentar, o que afetou negativamente o crescimento. Os resultados indicam a possibilidade de substituição de até 50 por cento do óleo de peixe por óleo de soja em dietas para juvenis do beijupirá.

The replacement of fish oil for cobia (Rachycentron canadum) in diets was evaluated using five proportions of fish oil:soybean oil. Two hundred juveniles (±12g) were randomly distributed in twenty 480L tanks and fed to apparent satiety twice daily for 42 days. Diets contained 12 percent lipids with different proportions of fish to soybean oils and were therefore named OP0, OP25, OP50, OP75 and OP100 according to the inclusion content of fish oil (0, 25, 50, 75 and 100 percent, respectively). Increasing dietary fish oil levels resulted in higher survival, growth and feed intake. Fish fed diets OP75 and OP100 presented higher weight gain than those fed diets OP0 and OP25, whereas fish fed diet OP50 had an intermediate weight gain. Overall, performance was enhanced at higher contents of dietary fish oil. No significant differences on protein and lipid content of fish carcass were observed, but moisture and ash content differed significantly between treatments. The inclusion of relatively higher contents of plant ingredients affected feed intake, which resulted in lower growth rates. The present results suggest the possibility of replacing up to 50 percent of fish oil with soybean oil in diets for cobia juveniles.

Moyamoya disease associated with renovascular disease in a young African-Brazilian patient.

Moyamoya is a rare disease characterized by fibrous dysplasia of the internal carotid and proximal cerebral arteries, which has been described mainly in young Japanese. We present a case of Moyamoya disease with renal artery involvement in a young male patient with an African origin. A 15-year-old boy was referred to our hospital due to uncontrolled blood pressure, headache, somnolence, cognitive deficit and multiple lacunar infarcts in the computed tomography. Cerebral arteriography showed the absence of the normal vascular anatomy at the level of the circle of Willis. The intracranial vessels presented severe stenosis or were occluded and replaced by an extensive network of ectasic collateral vessels. Abdominal ultrasound examination identified asymmetric kidneys, and renal arteriography showed a tight stenosis of the ostium and proximal segment of right main artery, which was only partially relieved by balloon angioplasty. A saphenous bypass from aorta to the right renal artery re-established the renal blood flow. Blood pressure dropped after surgery and was controlled with low doses of diuretic and beta-blocker. After arteriography he presented right hemiplegia, with partial recovering in the following months. In conclusion, we described the first case of Moyamoya disease with concomitant renovascular disease in a young adult of African origin, an association that may be more frequent than usually suspected in clinical practice.