Grb14 mRNA Levels During Follicular Deviation in Cattle are Higher in Granulosa Cells of Subordinate Compared to Dominant Follicles

The growth factor receptor‐bound protein 14 (Grb14) is a cellular adapter protein belonging to the Grb7 family of proteins. Studies with human and rodent cells have demonstrated that Grb14 acts as a negative regulator of tyrosine kinase receptor signalling through the MAPK and PI3K pathways. In cattle, tyrosine kinase receptors are activated during follicular development but the role of Grb14 in this process has not yet been investigated. Therefore, the aim of the present study was to characterize Grb14 mRNA expression in ovarian somatic cells during follicular growth and deviation in cattle. We found Grb14 mRNA expressed in both granulosa and theca cells derived from follicles at different stages of development (3–5 , 6–8, >8 mm in diameter). The abundance of mRNA for Grb14 was higher in granulosa cells of subordinate compared with those from dominant follicles at days 3 and 4 of the follicular wave (p < 0.05). Further, there was a negative correlation between the abundance of mRNA for Grb14 and P450Arom in granulosa cells (R² = 0.367; p < 0.05) and between the abundance of mRNA for Grb14 in granulosa cells and concentration of oestradiol in follicular fluid (R² = 0.545; p < 0.05). In theca cells, the expression of Grb14 mRNA did not differ between dominant and subordinate follicles (p > 0.05). These findings suggest that Grb14 may play a regulatory role in granulosa cells during follicular deviation in cattle.     

Sequential medium with GH and IGF‐1 improved in vitro development of bovine preantral follicles enclosed in ovarian tissue

The growth hormone (GH) and growth insulin‐like factor‐1 (IGF‐1) act directly upon the regulation and growth in the different phases of preantral follicles. Thus, it is necessary to define their sequentiality until the in vitro preovulatory development. Therefore, the study aimed to assess the effects of a sequential medium containing GH and/or IGF‐1 in the long‐duration in vitro culture of preantral ovarian follicles. Ovarian fragments were cultivated: first half (days 1–7), second half (days 7–14) or during 14 culture days. Treatments were identified as: αMEM+; GH → IGF‐1; IGF‐1 → GH and GH + IGF‐1. The culture was designed in 24‐well plates, in an incubator at 37°C and 5% CO₂. The parameters of normality, viability, follicles (primordial/in developing) and follicle diameter were evaluated. In addition, the ultrastructure was confirmed with electron transmission microscopy. The results showed that the culture treated with GH → IGF‐1 kept the follicular normality and the viability until the 14th day of culture and increased both in the follicular development until 7th day and in the follicular diameter until 14th day, when compared to the control. The treatments IGF‐1 → GH and GH + IGF‐1 were not effective in the developing and follicular diameter after 7 days of culture, and also reduced the percentage of viability. It is concluded that the bovine preantral follicles cultured in the sequential medium treated with GH → IGF‐1 improved the follicular development until the first half of the culture and kept these parameters with normality, viability and ultrastructure until the second half of the in vitro culture.     

<Emphasis Type="Italic">In vitro</Emphasis> morphogenic response of leaf sheath of <Emphasis Type="Italic">Phyllostachys bambusoides</Emphasis>

Nodal segments from secondary branches of saplings of Phyllostachys bambusoides were inoculated in MS medium to assess the in vitro morphogenic response of leaf sheath through the induction to callogenesis by Picloram(4-amino-3,5,6-trichloropicolinic acid) at different concentrations of carbohydrate under the same conditions with presence or absence of luminosity.In our experiment,secondary explants were kept in MS medium containing 8.0 mg·L-1 of Picloram for the callus formation.Calluses were transferred in MS medium supplemented with sucrose,fructose and glucose(control,2%,4% and 6%).Results show that Picloram induced the callogenesis in leaf sheath.The secondary embryogenesis was formed in yellow-globular callus.The sucrose as carbohydrate source in the absence of light was more efficient to induce rhizogenesis.Glucose was more efficiency in the presence of light.Callogenic induction and further embryogenesis evidenced the competence and determination of leaf sheath cells.     

Pathogenic and molecular comparison of Puccinia kuehnii isolates and reactions of sugarcane varieties to orange rust

Sugarcane orange rust, a disease caused by Puccinia kuehnii, was first reported in Brazil in 2009. There are no studies comparing the Brazilian P. kuehnii collections and the reaction of important sugarcane varieties under controlled conditions. This work compared the reaction of seven sugarcane varieties inoculated with six different P. kuehnii isolates from Brazilian sugarcane areas and verified the pathogenic and genetic variability of these isolates. The incubation (I) and latency (L) disease periods, disease severity (SEV), total number of lesions (TNL), total number of sporulating lesions (TNSL), and percentage of sporulating lesions (%SL) were evaluated. Furthermore, ITS1 and IGS ribosomal sequences of all P. kuehnii isolates used in this study were compared with pathogen sequences from 13 different countries. The disease incubation ranged from 7 to 10 days and the latency ranged from 10 to 21 days. SEV and TNL showed large variations and few significant differences between the reaction of the varieties to P. kuehnii, in contrast with the variables TNSL and %SL. The P. kuehnii isolates did not compose different virulent races, but the isolate from one site (Araras) was a more aggressive race. The ITS1 and IGS ribosomal sequences of six P. kuehnii isolates were identical with each other and to most P. kuehnii American sequences deposited at GenBank. The studied sequences of P. kuehnii isolates differed from the sequences from Asia, Tahiti and Oceania.     

Natriuretic peptide system regulation in granulosa cells during follicle deviation and ovulation in cattle

Natriuretic peptides (NPs) are known to regulate reproductive events in polyovulatory species, but their function and regulation in monovulatory species remain to be fully characterized. Using a well‐established in vivo model, we found that bovine granulosa cells from follicles near the deviation stage express mRNA for the three NP receptors (NPR1, NPR2 and NPR3), but not for NP precursors (NPPA, NPPB and NPPC). The abundance of NPR3 mRNA was higher in dominant compared to subordinate follicles at the expected time of follicular deviation. After deviation, mRNA for all NP receptors was significantly more abundant in the dominant follicle. Intrafollicular inhibition of oestrogen receptors downregulated NPR1 mRNA in dominant follicles. In granulosa cells from preovulatory follicles, NPPC mRNA increased at 3 and 6 h after systemic GnRH treatment, but decreased at 12 and 24 h to similar levels observed in samples collected at 0 h. After GnRH treatment, NPR1 mRNA was upregulated at 24 h, NPR3 mRNA gradually decreased after 3 h, while NPR2 mRNA was not regulated. The mRNA expression of the enzyme FURIN increased at 24 h after GnRH treatment. These findings revealed that the expression of mRNA encoding important components of the NP system is regulated in bovine granulosa cells during follicular deviation and in response to GnRH treatment, which suggests a role of NP system in the modulation of these processes in monovulatory species.     

Correlation between the nature of immunity induced by different immunogens and the establishment of latent infection by wild-type pseudorabies virus

To assess the correlation between the nature of immunity induced by different types of immunogens and the establishment of latent infection by wild-type pseudorabies virus (PrV), we used a murine model immunized with different immunogens, the PrV modified live vaccine (MLV), inactivated vaccine (IAV), and commercial oil-adjuvant subunit vaccine (OSV), via either intranasal (i.n.) or intramuscular (i.m.) route. Both MLV and IAV induced a different nature of immunity biased to Th1- and Th2-type, respectively, as judged by the ratio of PrV-specific IgG isotypes (IgG2a/IgG1) and the profile of cytokine IL-2, IL-4, and IFN-gamma production. In contrast, the OSV induced a lower isotype IgG2a to IgG1 ratio and higher level of IL-2 production. The MLV (inducing Th1-type) provided more effective protection against a virulent wild-type PrV challenge than IAV and OSV (inducing Th2- and mixed type, respectively). In addition, the MLV impeded the establishment of a latent infection with wild-type PrV, and the decrease in the PrV latency load by immunization with the MLV appeared to be mediated by the immune T-cells. These results demonstrate the substantial role of the immune responses driven by preceding vaccination in modulating the establishment of PrV latency caused by the post-infection of a field virus.     

Correlation of malt quality parameters and beer flavor stability: multivariate analysis

Malt is known to have an impact on beer flavor stability mainly due to the presence of antioxidants. In this study, five barley varieties were malted at industrial and micro scale, and quality parameters of the resulting malts were measured (diastatic power, friability, beta-glucan content, antiradical power, reducing power, lipoxygenase activity, and nonenal potential) and correlated with the sensory data obtained for the corresponding fresh and forced aged beers. A statistical strategy using multiple linear regressions was applied to explore relationships between the malt chemical parameters and beer sensory data, showing antiradical power as the major contribution of malt to beer flavor stability. Additionally, the measured antiradical power, which is well correlated with the polyphenolic content, was found to be very similar for malt and barley, emphasizing the key role of barley endogenous polyphenols.     

Impacts of anthropic pressures on soil phosphorus availability,concentration, and phosphorus forms in sediments in a Southern Brazilian watershed

Purpose  

The transfer of soil sediments and phosphorus from terrestrial to aquatic systems is a common process in agricultural lands. The aims of this paper are to quantify the soil phosphorus availability and to characterize phosphorus forms in soil sediments as contaminant agents of waters as a function of anthropic pressures.     

Selection of families and parents of sugarcane (Saccharum spp.) through mixed models by joint analysis of two harvests

At present, the sugarcane (Saccharum spp.) breeding programs from around the world have practiced early family selection, because in the early stages of breeding the evaluated characteristics have shown low heritability. This study aimed to select through restricted maximum likelihood/best linear unbiased prediction also known “mixed models” the best families among the 78 that comprise the RB05 sugarcane series, stage T1, of Programa de Melhoramento Genético da Cana-de-açúcar of the Rede Interuniversitária para o Desenvolvimento do Setor Sucroalcoleiro, Brazil. The experiments, conducted during 2007/2008 and 2008/2009 in Paranavaí, PR were implemented using an incomplete-block design with five replicates of each family. Each plot was composed of two planting rows of 5 m, spaced from each other by 1.40 m and spacing between plants was of 0.50 m, containing ten seedlings for planting row, resulting in a total of 20 individuals per replicate of each family. The characteristics evaluated were Brix (°Brix), ton of stalk per hectare (TCH) and ton of °Brix per hectare (TBH). The joint analysis obtained from these three traits in both harvests favored the selection of 35–41 families. The analysis also favored the selection of the best parents. The top five families for Brix were: F41M60, F02M77, F41M82, F61M38 and F62M35; to TCH were F66M30, F35M06, F78M45, F70M30 and F57M46 and for TBH the five elites families were F66M30, F35M06, F78M45, F70M30 and F01M39. The five best sugarcane parents for Brix were RB835486, SP91-1049, SP80-3280, RB72454 and RB925211; to TCH were IAC87-3396, RB941531, RB855511, RB915141 and RB957689, and for TBH the elite were IAC87-3396, RB855511, RB957689, RB941531 and RB855563. As each harvest season had different order of the best families, the joint analysis proved to be a primordial tool for plant breeders.