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This study aimed to evaluate the transcriptional changes occurring in isolated perfused mammary alveolar tissue in response to inoculation with S. agalactiae and to identify the most affected biological functions and pathways after 3 h. Four udders taken at slaughter from cows with healthy mammary gland were perfused ex situ with warmed and gassed Tyrode's solution. Mammary alveolar tissue samples were taken from the left fore and rear quarters (IQ-inoculated quarters) before inoculation (hour 0) and at 3 h post inoculation (hpi) and at the same times from control right fore and rear quarters (not inoculated: NIQ). A total of 1756 differentially expressed genes (DEGs) were identified between IQ and NIQ at 3 hpi using edgeR package. Within this set of DEGs, 952 were up regulated and mainly involved with innate immune response and inflammatory response, e.g., CD14, CCL5, TLR2, IL-8, SAA3, as well as in transcriptional regulation such as FOS, STAT3 and NFKBIA. Genes down-regulated (804) included those involved with lipid synthesis e.g., APOC2, SCD, FABP3 and FABP4. The most affected pathways were chemokine signaling, Wnt signaling and complement and coagulation cascades, which likely reflects the early stage response of mammary tissue to S. agalactiae infection. No significant gene expression changes were detected by RNA-Seq in the others contrasts. Real time-PCR confirmed the increase in mRNA abundance of immune-related genes: TLR2, TLR4, IL-1β, and IL-10 at 3 hpi between IQ and NIQ. The expression profiles of Casp1 and Bax for any contrasts were unaffected whereas Bcl2 was increased in IQ, which suggests no induction of apoptosis during the first hours after infection. Results provided novel information regarding the early functional pathways and gene network that orchestrate innate immune responses to S. agalactiae infection. This knowledge could contribute to new strategies to enhance resistance to this disease, such as genomic selection.
The uptake of genomic selection (GS) by the swine industry is still limited by the costs of genotyping. A feasible alternative to overcome this challenge is to genotype animals using an affordable low-density (LD) single nucleotide polymorphism (SNP) chip panel followed by accurate imputation to a high-density panel. Therefore, the main objective of this study was to screen incremental densities of LD panels in order to systematically identify one that balances the tradeoffs among imputation accuracy, prediction accuracy of genomic estimated breeding values (GEBVs), and genotype density (directly associated with genotyping costs). Genotypes using the Illumina Porcine60K BeadChip were available for 1378 Duroc (DU), 2361 Landrace (LA) and 3192 Yorkshire (YO) pigs. In addition, pseudo-phenotypes (de-regressed estimated breeding values) for five economically important traits were provided for the analysis. The reference population for genotyping imputation consisted of 931 DU, 1631 LA and 2103 YO animals and the remainder individuals were included in the validation population of each breed. A LD panel of 3000 evenly spaced SNPs (LD3K) yielded high imputation accuracy rates: 93.78% (DU), 97.07% (LA) and 97.00% (YO) and high correlations (>0.97) between the predicted GEBVs using the actual 60 K SNP genotypes and the imputed 60 K SNP genotypes for all traits and breeds. The imputation accuracy was influenced by the reference population size as well as the amount of parental genotype information available in the reference population. However, parental genotype information became less important when the LD panel had at least 3000 SNPs. The correlation of the GEBVs directly increased with an increase in imputation accuracy. When genotype information for both parents was available, a panel of 300 SNPs (imputed to 60 K) yielded GEBV predictions highly correlated (⩾0.90) with genomic predictions obtained based on the true 60 K panel, for all traits and breeds. For a small reference population size with no parents on reference population, it is recommended the use of a panel at least as dense as the LD3K and, when there are two parents in the reference population, a panel as small as the LD300 might be a feasible option. These findings are of great importance for the development of LD panels for swine in order to reduce genotyping costs, increase the uptake of GS and, therefore, optimize the profitability of the swine industry.
The present study investigated the impact of a western diet during gestation and lactation on the anthropometry, serum biochemical, blood pressure and cardiovascular autonomic control on the offspring. Male Wistar rats were divided into two groups according to their mother’s diet received: control group (C: 18% calories of lipids) and westernized group (W: 32% calories of lipids). After weaning both groups received standard diet. On the 60th day of life, blood samples were collected for the analysis of fasting glucose and lipidogram. Cardiovascular parameters were measured on the same period. Autonomic nervous system modulation was evaluated by spectrum analysis of heart rate (HR) and systolic arterial pressure (SAP). The W increased glycemia (123±2 v. 155±2 mg/dl), low-density lipoprotein (15±1 v. 31±2 mg/dl), triglycerides (49±1 v. 85±2 mg/dl), total cholesterol (75±2 v. 86±2 mg/dl), and decreased high-density lipoprotein (50±4 v. 38±3 mg/dl), as well as increased body mass (209±4 v. 229±6 g) than C. Furthermore, the W showed higher SAP (130±4 v. 157±2 mmHg), HR (357±10 v. 428±14 bpm), sympathetic modulation to vessels (2.3±0.56 v. 6±0.84 mmHg2) and LF/HF ratio (0.15±0.01 v. 0.7±0.2) than C. These findings suggest that a western diet during pregnancy and lactation leads to overweight associated with autonomic misbalance and hypertension in adulthood.
The aim of this study was to evaluate the effect of dietary lysine on performance, protein deposition and respiratory chain gene expression in male broilers. A total of 252 Cobb 500 broilers were distributed, in a completely randomized design, into four treatments with seven replicates of nine birds per experimental unit. Experimental treatments consisted of diets based on corn and soybean meal, with four levels of digestible lysine: 1.016%, 1.099%, 1.182% and 1.265%. The increase in the level of digestible lysine in the diet provided higher weight gains, feed efficiency and body protein deposition. Birds fed the lowest level of dietary lysine (1.016%) showed a lower expression of genes such as NADH dehydrogenase subunit I (ND1), cytochrome b (CYTB) and cytochrome c oxidase subunits I (COX I), II (COX II) and III (COX III), displaying the worst performance and body protein deposition. This demonstrates the relationship existing between the expression of the evaluated genes and the performance responses. In conclusion, results indicate that broilers fed diets with higher levels of digestible lysine have increased messenger RNA expression of some genes coded in the mitochondrial electron transport chain (ND1, CYTB, COX I, COX II and COX III). It may be stated that diets with proper levels of digestible lysine, within the ‘ideal protein’ concept, promote the expression of genes, which increases the mitochondrial energy, thereby fostering body protein deposition and the performance of broilers in the starter phase.
The dune of Oitavos, the underlying paleosol, and Helix sp. gastropod shells found within the paleosol were dated using a combination of radiocarbon and blue optically stimulated luminescence (OSL). The organic component of the paleosol produced a significantly older age (∼20,000 cal BP) than the OSL age measurement (∼15,000 yr), while 14C age measurements on the inorganic component and the gastropods produced ages of ∼35,000 yr and ∼34,000 yr, respectively. Rare-earth element analyses provide evidence that the gastropods incorporate geological carbonate, making them an unreliable indicator of the age of the paleosol. We propose that the 14C age of the small organic component of the paleosol is also likely to be unreliable due to incorporation of residual material. The OSL age measurement of the upper paleosol (∼15,000 yr) is consistent with the age for the base of the dune (∼14,500 yr). The younger OSL age for the top of the dune (∼12,000 yr) suggests that it was built up by at least 2 sand pulses or that there was a remobilization of material at the top during its evolution, prior to consolidation.
The objective of the current study was to assess the use of nonlinear mixed model methodology to fit the growth curves (weight v. time) of two dairy goat genotypes (Alpine, +A and Saanen, +S). The nonlinear functions evaluated included Brody, Von Bertalanffy, Richards, Logistic and Gompertz. The growth curve adjustment was performed using two steps. First, random effects u1, u2 and u3 were linked to the asymptotic body weight (β1), constant of integration (β2) and rate constant of growth (β3) parameters, respectively. In addition to a traditional fixed-effects model, four combinations of models were evaluated using random variables: all parameters associated with random effects (u1, u2 and u3), only β1 and β2 (u1 and u2), only β1 and β3 (u1 and u3) and only β1 (u1). Second, the fit of the best adjusted model was refined by using the power variance and modelling the error structure. Residual variance (
$\sigma _e^2 $
) and the Akaike information criterion were used to evaluate the models. After the best fitting model was chosen, the genotype curve parameters were compared. The residual variance was reduced in all scenarios for which random effects were considered. The Richards (u1 and u3) function had the best fit to the data. This model was reparameterized using two isotropic error structures for unequally spaced data, and the structure known in the literature as SP(MATERN) proved to be a better fit. The growth curve parameters differed between the two genotypes, with the exception of the constant that determines the proportion of the final size at which the inflection point occurs (β4). The nonlinear mixed model methodology is an efficient tool for evaluating growth curve features, and it is advisable to assign biologically significant parameters with random effects. Moreover, evaluating error structure modelling is recommended to account for possible correlated errors that may be present even when using random effects. Different Richard growth curve parameters should be used for the predominantly Alpine and Saanen genotypes because there are differences in their growth patterns.
Children with conduct problems (CP) are a heterogeneous group. Those with high levels of callous–unemotional traits (CP/HCU) appear emotionally under-reactive at behavioural and neural levels whereas those with low levels of CU traits (CP/LCU) appear emotionally over-reactive, compared with typically developing (TD) controls. Investigating the degree to which these patterns of emotional reactivity are malleable may have important translational implications. Instructing participants with CP/HCU to focus on the eyes of fearful faces (i.e. the most salient feature) can ameliorate their fear-recognition deficits, but it is unknown whether this is mediated by amygdala response. It is also unknown whether focusing on fearful eyes is associated with increased amygdala reactivity in CP/LCU.
Functional magnetic resonance imaging (fMRI) was used to measure neural responses to fearful and calm faces in children with CP/HCU, CP/LCU and TD controls (n = 17 per group). On half of trials participants looked for a blue dot anywhere within target faces; on the other half, participants were directed to focus on the eye region.
Reaction time (RT) data showed that CP/LCU were selectively slowed in the fear/eyes condition. For the same condition, CP/LCU also showed increased amygdala and subgenual anterior cingulate cortex (sgACC)/orbitofrontal cortex (OFC) responses compared with TD controls. RT and amygdala response to fear/eyes were correlated in CP/LCU only. No effects of focusing on the eye region were observed in CP/HCU.
These data extend the evidence base suggesting that CU traits index meaningful heterogeneity in conduct problems. Focusing on regulating reactive emotional responses may be a fruitful strategy for children with CP/LCU.
The effect of feed restriction on gene expression of regulatory enzymes of intermediary metabolism was studied in two sheep breeds (Australian Merino and Dorper) subjected to two nutritional treatments: feed restriction (85% of daily maintenance requirements) and control (ad libitum feeding), during 42 days. The experimental animals (ram lambs) were divided into four groups, n = 5 (Australian Merino control (MC), Australian Merino Restriction (MR), Dorper control (DC) and Dorper Restriction (DR)). After the trial, animals were sacrificed and samples were taken from liver tissue to quantify glucose levels and gene expression of relevant intermediary metabolism enzymes (phosphofructokinase (PFK), pyruvate kinase (PK), phosphoenolpyruvate carboxykinase, fructose 1,6-bisphosphatase, glucose-6-phosphatase, glycogen synthase (GS), fatty acid synthase (FAS), glutamate dehydrogenase (GDH) and carbamoyl phosphate synthase (CPS)) through real-time PCR. During the experimental period, the MR animals lost 12.6% in BW compared with 5.3% lost by the Dorper lambs. MC and DC rams gained, respectively, 8.8% and 14% during the same period. Within the Dorper breed, restricted feed animals revealed a significant decrease over controls in the transcription of PFK (1.95-fold) and PK (2.26-fold), both glycolytic enzymes. The gluconeogenesis showed no change in the feed restricted animals of both breeds. DR feed group presented a significant decrease over the homologous Merino sheep group on GS. In both experimental breeds, FAS mRNA expression was decreased in restricted feed groups. GDH expression was decreased only in the DR animals (1.84-fold) indicating a reduced catabolism of amino acids in these animals. Finally, CPS was significantly (P < 0.05) higher in the Dorper sheep, indicating a facilitated urea synthesis in this breed. These results indicate a better adaptation of metabolic intermediate regulatory enzymes and hepatic glucose production of Dorper sheep to feed restriction concurring with the BW results in the experimental groups.
In this work we present two aspects of the Astronomy education activities carried out in 2012 by a multidisciplinary group at Universidad de Guanajuato, including specialists in Astronomy, Social Sciences and Environmental Engineering. The first program linked the Venus Transit, occurred in June 2012, with a national campaign of vulgarization of both modern and ancient (Mayan) Astronomy. Professional astronomers all around the country took advantage of the recent myth linked to the end of a large Mayan calendar cycle (13 baktuns, or some 5125 years) happening, after certain authors, in December 2012. In Guanajuato, the Astronomy Department organized live observations of the Venus Transit at two different locations, and complemented with conferences about astronomical events and the fake predictions of disasters linked to the “end“ of the Mayan calendar. This program was very successful not only in Guanajuato but throughout the country, with several thousands of people attending live observations, conferences, expositions, etc.
The efficiency of in vitro fertilization (IVF) depends on the viability of spermatozoa. For capuchin monkeys (Cebus apella), in vitro capacitation of spermatozoa is challenging because of their unique seminal coagulum. Motile spermatozoa can be obtained after liquefaction of the semen coagulum in coconut water-based solution. The objective of the present study was to establish an optimal in vitro maturation (IVM) protocol for capuchin monkeys and to observe the effect of follicle stimulating hormone (FSH) and luteinising hormone (LH) on IVF and parthenogenetic activation (PA) of oocytes collected from unstimulated females. We assessed spermatozoa quality after recovery from seminal coagulum using the solution ACP-118® as an extender. Oocytes were matured in vitro for 36 or 40 h and subjected to IVF or PA by applying ionomycin combined either with 6-dimethylaminopurine (6-DMAP) or roscovitine. In total, 87% of oocytes reached metaphase II (MII) after 40 IVM and 4-cell embryo production was obtained after IVF and parthenogenesis using ionomycin/6-DMAP. ACP-118® was used successfully to harvest viable spermatozoa from semen coagulum and in the preservation of spermatozoa, which were able to fertilize oocytes in vitro.
Silica doped alumina/aluminate materials present a combination of high strength and high toughness not achieved before in other alumina systems, except for transformation toughened alumina. We have associated the increase in toughness to crack bridging by anisotropically grown alumina grains with concurrent interfacial debonding of these grains. A HREM study of grain boundaries and hetero-interface structures in this material shows the absence of amorphous phases at grain boundaries. Local Auger electron analysis of fractured surfaces revealed the coexistence of Si and La at the grain facets exposed by the noticeable intergranular fracture mode of this material. It is concluded that a certain and important degree of boundaries weakness is related to both, presence of Si at the interfaces and existence of alumina/aluminate hetero-interfaces.
Intergranular glass phases can have a significant influence on the fracture resistance (R-curve behavior) of silicon nitride ceramics and appears to be related to the debonding of the β-Si3N4/oxynitride-glass interfaces. Applying the results from β-Si3N4-whisker/oxynitride-glass model systems, self-reinforced silicon nitrides with different sintering additive ratios were investigated. Silicon nitrides sintered with a lower Al2O3:Y2O3 additive ratio exhibited higher steady-state fracture toughness together with a steeply-rising R-curve. Analytical electron microscopy studies suggested that the different fracture behavior is related to the Al content in the SiAlON growth band on the elongated grains, which could result in differences in interfacial bonding structures between the grains and the intergranular glass.
We present experimental studies on the interaction of soft X-rays on gas-phase and solid-phase amino acids and nucleobases in an attempt to verify if these molecules (supposed to be formed in molecular clouds/protostellar clouds) can survive long enough to be observed or even to be found in meteorites. Measurements have been undertaken employing 150 eV photons under high vacuum conditions at the Brazilian Synchrotron Light Laboratory (LNLS). The produced ions from the gas-phase experiments (glycine, adenine and uracil) have been mass/charge analyzed by time-of-flight spectrometer. The analysis of solid phase samples (glycine, DL-proline, DL-valine, adenine and uracil) were performed by a Fourier transform infrared spectrometer coupled to the experimental chamber. Photodissociation cross sections and halflives were determined and extrapolated to astrophysical environments. The nucleobases photostability was up to two orders of magnitude higher than for the amino acids.
Aeroengine components operate under particularly severe conditions, including the combination of both high mechanical stresses and elevated temperatures in chemically aggressive environments. These normal operational requirements will eventually promote both cyclic and time dependent failure mechanisms related with creep-fatigue damages. Therefore, the use of adequate preventive inspection techniques is mandatory in order to maximize safety and reliability levels of these components during service. However, there are some occasional and unpredictable material failures which must be conveniently investigated in order to avoid future hazardous situations. Scanning electron microscopy (SEM) can play a major role in the context of evaluation of failed components, allowing the observation of fracture surfaces and other material's features which are essential to the identification of some particular failure mechanisms.
We present experimental studies on the photoionization and photodissociation processes (photodestruction) of gaseous amino acids and nucleobases in interstellar and interpla-netary radiation analogs conditions. The measurements have been undertaken at the Brazilian Synchrotron Light Laboratory (LNLS), employing vacuum ultraviolet (VUV) and soft X-ray photons. The experimental set up basically consists of a time-of-flight mass spectrometer kept under high vacuum conditions. Mass spectra were obtained using a photoelectron photoion coincidence technique. We have shown that the amino acids are effectively more destroyed (up to 70–80%) by the stellar radiation than the nucleobases, mainly in the VUV. Since polycyclic aromatic hydrocarbons have the same survival capability and seem to be ubiquitous in the ISM, it is not unreasonable to predict that nucleobases could survive in the interstellar medium and/or in comets, even as a stable cation.
This study provides the first compilation on age and growth of some delphinids in south-eastern Brazil (18°25′S–25°45′S). A total of 154 delphinids were reported: 44 Atlantic spotted dolphin Stenella frontalis; 36 bottlenose dolphin Tursiops truncatus; 26 ‘Brazilian’ common dolphin Delphinus sp.; 20 rough-toothed dolphin Steno bredanensis; 16 Fraser's dolphin Lagenodelphis hosei; 3 false killer whale Pseudorca crassidens; 3 unidentified Stenella sp.; 2 pantropical spotted dolphin Stenella attenuata; 2 short-finned pilot whale Globicephala macrorhynchus; 1 spinner dolphin Stenella longirostris; and 1 striped dolphin Stenella coeruleoalba. Age was estimated by counting the number of growth layer groups present in the dentine in 74.5% of the sample. The growth of 92 individuals of the first five species was determined by the Gompertz model to length-at-age data. Stenella frontalis—the oldest specimen was 23 y and the asymptotic length of 224.4 cm predicted by growth curve occurred at about 20 y; T. truncatus—the oldest specimen was 26 y and the asymptotic length of 301.3 cm predicted by growth curve occurred at about 20 y; Delphinus sp.—the oldest dolphin was 18 y and the asymptotic length of 215.9 cm predicted by growth curve occurred at about 5–6 y; S. bredanensis—the oldest specimen was 24 y and the asymptotic length of 258.1 cm predicted by growth curve occurred at about 10 y; L. hosei—the oldest specimen was 19 y and the asymptotic length of 231.2 cm predicted by growth curve occurred at about 7–8 y. Only age was estimated for the other species. The age-at-length data for S. frontalis, Delphinus sp., S. bredanensis and L. hosei were consistent, suggesting a good agreement with previous work on these species. For T. truncatus, the age at asymptotic length obtained in this study might be confirmed by increasing the sample size. The information currently presented will contribute to further life history research of delphinids on the western south Atlantic coast.