Early morphological and neurochemical changes of the bed nucleus of stria terminalis (BNST) in gestational protein-restricted male offspring.

BACKGROUND: The bed nucleus of the stria terminalis (BNST) is a structure with a peculiar neurochemical composition involved in modulating anxietylike behavior and fear. AIM: The present study investigated the effects on the BNST neurochemical composition and neuronal structure in critical moments of the postnatal period in gestational protein-restricted male rats' offspring. METHODS: Dams were maintained during the pregnancy on isocaloric rodent laboratory chow with standard protein content [NP, 17%] or low protein content [LP, 6%]. BNST from male NP and age-matched LP offspring was studied using the isotropic fractionator method, Neuronal 3D reconstruction, dendritic-tree analysis, blotting analysis, and high-performance liquid chromatography. RESULTS: Serum corticosterone levels were higher in male LP offspring than NP rats in 14-day-old offspring, without any difference in 7-day-old progeny. The BNST total cell number and anterodorsal BNST division volume in LP progeny were significantly reduced on the 14th postnatal day compared with NP offspring. The BNST HPLC analysis from 7 days-old LP revealed increased norepinephrine levels compared to NP progeny. The BNST blot analysis from 7-day-old LP revealed reduced levels of GR and BDNF associated with enhanced CRF1 expression compared to NP offspring. 14-day-old LP offspring showed reduced expression of MR and 5HT1A associated with decreased DOPAC and DOPA turnover levels relative to NP rats. In Conclusion, the BNST cellular and neurochemical changes may represent adaptation during development in response to elevated fetal exposure to maternal corticosteroid levels. In this way, gestational malnutrition alters the BNST content and structure and contributes to already-known behavioral changes.

Effect of supplementation with algae ß-glucans on performance, health, and blood metabolites of Holstein dairy calves.

Studies have shown that ß-glucans extracted from the cell wall of cereals, algae, and yeasts have been associated with improved immune function. However, it is unknown whether algae ß-glucan supplementation affects the performance, blood metabolites, or cell counts of immune cells in dairy calves. The objective of this randomized clinical trial was to evaluate whether supplementation of ß-glucans to milk replacer in dairy calves fed 6 L/d improved growth performance and fecal status and altered the blood metabolite profile. In this trial, we enrolled Holstein calves (n = 34) at birth (body weight 36.38 ± 1.33 kg; mean ± standard deviation) to receive, from 1 d of age, either 2 g/d algae ß-glucans mixed into 6 L/d of milk replacer (22.4% crude protein and 16.2% fat) or an unsupplemented milk replacer (control). The calves were blocked in pairs according to birth weight, sex, and date of birth (up to 5 d difference). Calves were housed individually, and calf starter (24.7% crude protein and 13.9% neutral detergent fiber) was offered ad libitum based on orts of the previous day until 56 d of age (end of the trial). Body weight was measured weekly, and health checks and daily fecal consistency were evaluated daily in every calf by the same observer. Calves with 2 consecutive days of loose feces that sifted through bedding were considered diarrhea positive. We used a linear mixed effects model to evaluate the effects of ß-glucan supplementation fed during the preweaning period on performance (average daily gain), final weight, feed efficiency (FE), white blood cell count, and selected blood metabolites, repeated by time. A generalized linear mixed effects model was also run to evaluate the likelihood of a diarrhea bout in the first 28 d of life, controlling for the calf as the subject with a logistic distribution. We included age, serum total protein at 48 h, and birth weight as covariates. At 56 d, ß-glucan-supplemented calves weighed more than control calves (56.3 vs. 51.5 kg). Treatment had no effect on total starter intake, but there was a treatment by age interaction for FE, with greater FE for ß-glucan-supplemented calves in wk 3 and 5 of age. There was only a tendency for average daily gain to be greater in supplemented calves than in control calves for the duration of the study. Furthermore, control calves had 14.66 [95% confidence interval (95% CI): 9.87-21.77] times greater odds of having a diarrheal bout than ß-glucan-supplemented calves. Control calves had 12.70 (95% CI: 8.82-18.28) times greater odds of having an additional day with an abnormal fecal score compared with ß-glucan-supplemented calves, suggesting that supplementation ameliorated diarrhea severity. We found no association of treatment with concentrations of serum total protein, albumin, creatinine, or glucose during the preweaning period. Our findings suggest that dietary supplementation of 2 g/d of algae ß-glucans to milk replacer improved fecal status and may affect growth, as evidenced by a higher weaning weight, compared with control calves. Future studies should explore the effect of algae ß-glucans on lower-gut physiology and digestibility in dairy calves.

Supplementation of lysolecithin in milk replacer for Holstein dairy calves: Effects on growth performance, health, and metabolites.

Lysolecithin is an antiinflammatory emulsifier associated with improved apparent digestibility of total dietary fat and improved feed efficiency in dairy cattle. However, it is unknown if lysolecithin (LYSO) improves performance in calves. Moreover, since many conventional milk replacers use vegetable-sourced fat (e.g., palm oil), nutrient absorption and fecal score may be affected in neonatal calves. Thus, the objective of this study was to evaluate the effects of LYSO supplemented in milk replacer on performance, metabolites, and gut health of preweaned dairy calves. Holstein calves (n = 32) with adequate passive transfer were assigned in pairs (16 blocks) balanced by birth weight, date of birth, and sex at 1 d of age to randomly receive either LYSO (mixed in 2 milk replacer feedings at a rate of 4 g/d Lysoforte, Kemin Industries Inc., Des Moines, IA) or a milk replacer control (nothing added). Both treatments were fed 6 L/d milk replacer [22.5% crude protein, 16.2% crude fat (vegetable oil fat source) on a dry matter basis with 14% solids] by bucket in 2 daily feedings for 56 d. Calves were individually housed in wooden hutches and offered a commercial calf starter (24.6% crude protein and 13.9% neutral detergent fiber) and water by bucket ad libitum. Feed refusals and calf health was assessed daily. Weights and blood metabolites (glucose, total serum protein, albumin, creatinine, triglycerides, and cholesterol) were sampled weekly, and calves completed the study before weaning at 56 d of age. The effects of LYSO on calf average daily gain, feed efficiency, and blood metabolites were evaluated using a linear mixed model with time as a repeated measure, calf as the subject, and block as a random effect in SAS (SAS Institute Inc., Cary, NC). The effect of LYSO to improve the odds of abnormal fecal score was evaluated using a logistic model. Supplementation of LYSO increased average daily gain (control 0.28 ± 0.03 kg; LYSO 0.37 ± 0.03 kg; least squares means ± standard error of the mean) and increased feed efficiency (gain-to-feed; control 0.25 ± 0.03; LYSO 0.32 ± 0.03). Similarly, LYSO calves had a higher final body weight at d 56 (control 52.11 ± 2.33 kg; LYSO 56.73 ± 2.33 kg). Interestingly, total dry matter intake was not associated with LYSO despite improved average daily gain (total dry matter intake control 1,088.7 ± 27.62 g; total dry matter intake LYSO 1,124.8 ± 27.62 g). Blood glucose, albumin, creatinine, triglycerides, and cholesterol were not associated with LYSO. Indeed, only total serum protein had a significant interaction with LYSO and age at wk 5 and 6. Moreover, control calves had a 13.57 (95% confidence interval: 9.25-19.90) times greater odds of having an abnormal fecal score on any given day during the diarrhea risk period from d 1 to 28. The inclusion of LYSO as an additive in milk replacer in a dose of 4 g/d may improve performance, and calf fecal score, preweaning. Further research should investigate the mechanisms behind the effects of LYSO on fat digestibility in calves fed 6 L/d of milk replacer with vegetable-sourced fat.

Climate-induced variability in South Atlantic wave direction over the past three millennia.

Through alteration of wave-generating atmospheric systems, global climate changes play a fundamental role in regional wave climate. However, long-term wave-climate cycles and their associated forcing mechanisms remain poorly constrained, in part due to a relative dearth of highly resolved archives. Here we use the morphology of former shorelines preserved in beach-foredune ridges (BFR) within a protected embayment to reconstruct changes in predominant wave directions in the Subtropical South Atlantic during the last ~ 3000 years. These analyses reveal multi-centennial cycles of oscillation in predominant wave direction in accordance with stronger (weaker) South Atlantic mid- to high-latitudes mean sea-level pressure gradient and zonal westerly winds, favouring wave generation zones in higher (lower) latitudes and consequent southerly (easterly) wave components. We identify the Southern Annular Mode as the primary climate driver responsible for these changes. Long-term variations in interhemispheric surface temperature anomalies coexist with oscillations in wave direction, which indicates the influence of temperature-driven atmospheric teleconnections on wave-generation cycles. These results provide a novel geomorphic proxy for paleoenvironmental reconstructions and present new insights into the role of global multi-decadal to multi-centennial climate variability in controlling coastal-ocean wave climate.

Red propolis as an additive for preweaned dairy calves: Effect on growth performance, health, and selected blood parameters.

Propolis is a natural product produced by bees and sold commercially. It is well known for its antioxidant and anti-inflammatory effects and the ability to improve immune system functions in humans and animals. Many of its positive effects can contribute positively to animal productivity. The aim of this study was to evaluate possible beneficial effects of red propolis supplementation on growth performance, metabolism, and health status of dairy calves during the preweaning phase. Thirty-two newborn calves were individually housed, with free access to water and concentrate, and fed 6 L/d of whole milk. Animals were distributed in 2 treatments: (1) control and (2) supplementation with 4 mL/d of red propolis ethanolic extract (30%; EEP) in whole milk. Feed consumption and fecal and health scores were monitored daily from 1 d of age to 56 d. Calves were weighed and measured weekly. Blood samples were collected weekly, 2 h after morning milk feeding, for determination of glucose, total serum protein, urea, lactate, ß-hydroxybutyrate, total iron binding capacity, and total leukocytes (white blood cells) and erythrocytes (red blood cells). Starter feed intake, daily weight gain, and body measurements were not affected by the EEP supplementation. Blood parameters were also not affected, with the exception for the red blood cell counts, which were lower in animals supplemented with EEP. Supplementation with EEP significantly decreased fecal score, days with diarrhea, and veterinary treatments of diarrhea. These results indicate that red propolis supplementation has potential to improve calf health and reduce the incidence of diarrhea and, as a consequence, the use of antibiotics in calf rearing systems.

Direct inhibitory effect of caffeine on viability, synthesis activity and gene expression in cultures of chondrocytes extracted from the articular cartilage of rats / Efeito inibidor direto da cafeína sobre a viabilidade, atividade de síntese e expressão gênica em culturas de condrócitos da cartilagem articular de ratos

The aim of this study was to evaluate the effect of concentrations of caffeine on the viability, synthesis activity and gene expression in cultures of chondrocytes. Extracted articular cartilage from the femurs and tibias of 15 Wistar rats at three days old to isolate chondrocytes. Chondrocytes were cultured in chondrogenic medium (control) or supplemented with caffeine (0.5, 1.0, 2.0mM). Cell viability, alkaline phosphatase activity and collagen synthesis were assessed using colorimetric assays at 7, 14, 21 days. The chondrocyte cultures of all groups grown under coverslips were stained with hematoxylin-eosin to determine the percentage of cells/field and with PAS, safranin O, alcian blue to determine the percentage of matrix chondrogenic/field at 21 days. The expressions of gene transcripts for aggrecan, collagen-II, Sox-9, Runx-2 and alkaline phosphatase were also evaluated by RT-PCR at 21 days. The means were compared using Student-Newman-Keuls. Caffeine significantly reduced the conversion of MTT to formazan, percentage of cells/field, collagen synthesis, alkaline phosphatase activity, synthesis of PAS+, safranin O+ and alcian blue+ chondrogenic matrix, and the expression of aggrecan, Sox-9 and II collagen. It is concluded that caffeine at concentrations of 0.5, 1.0, 2.0mM has a direct inhibitory effect on chondrogenesis in cultures of chondrocytes from rats.(AU)

O objetivo deste estudo foi avaliar o efeito direto de concentrações de cafeína sobre a viabilidade, atividade de síntese e expressão gênica em culturas de condrócitos de ratos. As cartilagens dos fêmures e tíbias de 15 ratos Wistar com três dias foram extraídas para isolamento de condrócitos. Os condrócitos foram cultivados em meio condrogênico (controle) ou em meio acrescido de diferentes concentrações de cafeína (0,5, 1,0, 2,0mM). Foram avaliadas a viabilidade celular, a atividade da fosfatase alcalina e a síntese de colágeno por ensaios colorimétricos aos sete, 14 e 21 dias. Condrócitos cultivados sob lamínulas foram corados pela hematoxilina e eosina, para se determinar a porcentagem de células/campo, e pelo PAS, safranina O, alcian Blue, para se determinar a porcentagem de matriz condrogênica/campo aos 21 dias. Foi avaliada a expressão de transcriptos gênicos para Sox-9, Runx-2, agrecano, colágeno-II e fosfatase alcalina por qRT-PCR, aos 21 dias. As médias foram comparadas pelo Student-Newman-Keuls. A cafeína reduziu significativamente o MTT em cristais de formazan, a porcentagem de células/campo, a síntese de colágeno, a atividade da fosfatase alcalina e a síntese de matriz condrogênica PAS+, safranina O+, alcian blue+ e expressão de Sox-9 e colágeno-II. Conclui-se que a cafeína, nas concentrações de 0,5, 1,0, 2,0mM, apresenta efeito inibidor direto sobre a condrogênese em culturas de condrócitos de ratos.(AU)

Direct inhibitory effect of caffeine on viability, synthesis activity and gene expression in cultures of chondrocytes extracted from the articular cartilage of rats / Efeito inibidor direto da cafeína sobre a viabilidade, atividade de síntese e expressão gênica em culturas de condrócitos da cartilagem articular de ratos

The aim of this study was to evaluate the effect of concentrations of caffeine on the viability, synthesis activity and gene expression in cultures of chondrocytes. Extracted articular cartilage from the femurs and tibias of 15 Wistar rats at three days old to isolate chondrocytes. Chondrocytes were cultured in chondrogenic medium (control) or supplemented with caffeine (0.5, 1.0, 2.0mM). Cell viability, alkaline phosphatase activity and collagen synthesis were assessed using colorimetric assays at 7, 14, 21 days. The chondrocyte cultures of all groups grown under coverslips were stained with hematoxylin-eosin to determine the percentage of cells/field and with PAS, safranin O, alcian blue to determine the percentage of matrix chondrogenic/field at 21 days. The expressions of gene transcripts for aggrecan, collagen-II, Sox-9, Runx-2 and alkaline phosphatase were also evaluated by RT-PCR at 21 days. The means were compared using Student-Newman-Keuls. Caffeine significantly reduced the conversion of MTT to formazan, percentage of cells/field, collagen synthesis, alkaline phosphatase activity, synthesis of PAS+, safranin O+ and alcian blue+ chondrogenic matrix, and the expression of aggrecan, Sox-9 and II collagen. It is concluded that caffeine at concentrations of 0.5, 1.0, 2.0mM has a direct inhibitory effect on chondrogenesis in cultures of chondrocytes from rats.(AU)

O objetivo deste estudo foi avaliar o efeito direto de concentrações de cafeína sobre a viabilidade, atividade de síntese e expressão gênica em culturas de condrócitos de ratos. As cartilagens dos fêmures e tíbias de 15 ratos Wistar com três dias foram extraídas para isolamento de condrócitos. Os condrócitos foram cultivados em meio condrogênico (controle) ou em meio acrescido de diferentes concentrações de cafeína (0,5, 1,0, 2,0mM). Foram avaliadas a viabilidade celular, a atividade da fosfatase alcalina e a síntese de colágeno por ensaios colorimétricos aos sete, 14 e 21 dias. Condrócitos cultivados sob lamínulas foram corados pela hematoxilina e eosina, para se determinar a porcentagem de células/campo, e pelo PAS, safranina O, alcian Blue, para se determinar a porcentagem de matriz condrogênica/campo aos 21 dias. Foi avaliada a expressão de transcriptos gênicos para Sox-9, Runx-2, agrecano, colágeno-II e fosfatase alcalina por qRT-PCR, aos 21 dias. As médias foram comparadas pelo Student-Newman-Keuls. A cafeína reduziu significativamente o MTT em cristais de formazan, a porcentagem de células/campo, a síntese de colágeno, a atividade da fosfatase alcalina e a síntese de matriz condrogênica PAS+, safranina O+, alcian blue+ e expressão de Sox-9 e colágeno-II. Conclui-se que a cafeína, nas concentrações de 0,5, 1,0, 2,0mM, apresenta efeito inibidor direto sobre a condrogênese em culturas de condrócitos de ratos.(AU)

Beak trimming by infrared radiation of layers

This study aimed at evaluating beak-trimming methods and intensities on the performance, beak length and uniformity, and egg parameters of commercial layers. During the rearing period, birds were distributed in randomized blocks, with a 2x3 factorial design + two treatments, consisting of beak trimming with infrared (IR) radiation (moderate or severe); three IR intensities (low: 42 nm, intermediate: 46 nm, or high: 52 nm); and two control hot blade (HB) beak trimming treatments (moderate or severe) with eight treatments and five replicates of 21 chicks each, totaling 840 birds. During the growing phase (second experiment) the factorial design was modified to 2 x 3 x 2 + 4, comprising two IR beak-trimming intensities (moderate or severe); three IR radiation intensities (low, moderate and high), second beak trimming at 11 weeks of age or not, and four additional HB beak-trimming treatments (moderate or severe) beak-trimming intensities, and second beak trimming at 11 weeks of age or not. There were 16 treatments with six replicates of seven birds each, totaling 672 laying hens, out of which 576 were selected for evaluation during the production period, with the same number of treatments and replicates of the previous phase. Weight gain (g), body weight (g), body weight uniformity (%), beak length (mm) and uniformity (%), absence of cannibalism (%), livability (%), egg production (%), feed intake (g), feed conversion per eggmass, eggmass, specific gravity, eggshell strength, Haugh unit, % yolk were evaluated. Beak trimming by infra-red radiation may be employed in commercial laying hens.

Beak trimming by infrared radiation of layers

This study aimed at evaluating beak-trimming methods and intensities on the performance, beak length and uniformity, and egg parameters of commercial layers. During the rearing period, birds were distributed in randomized blocks, with a 2x3 factorial design + two treatments, consisting of beak trimming with infrared (IR) radiation (moderate or severe); three IR intensities (low: 42 nm, intermediate: 46 nm, or high: 52 nm); and two control hot blade (HB) beak trimming treatments (moderate or severe) with eight treatments and five replicates of 21 chicks each, totaling 840 birds. During the growing phase (second experiment) the factorial design was modified to 2 x 3 x 2 + 4, comprising two IR beak-trimming intensities (moderate or severe); three IR radiation intensities (low, moderate and high), second beak trimming at 11 weeks of age or not, and four additional HB beak-trimming treatments (moderate or severe) beak-trimming intensities, and second beak trimming at 11 weeks of age or not. There were 16 treatments with six replicates of seven birds each, totaling 672 laying hens, out of which 576 were selected for evaluation during the production period, with the same number of treatments and replicates of the previous phase. Weight gain (g), body weight (g), body weight uniformity (%), beak length (mm) and uniformity (%), absence of cannibalism (%), livability (%), egg production (%), feed intake (g), feed conversion per eggmass, eggmass, specific gravity, eggshell strength, Haugh unit, % yolk were evaluated. Beak trimming by infra-red radiation may be employed in commercial laying hens.(AU)

Production indicators and levels of corticosterone in pullets treated with beak-trimming protocols

The aim of this study was to compare the effects of beak trimming by hot blade or infrared radiation on production indicators and plasma levels of corticosterone in pullets. During the rearing phase, an entirely random delineation was used a 2 x 3 factorial design (two different techniques for beak trimming x 3 strains), a total of six treatments with six repetitions of 20 hens. In the growing phase, the previous treatments were divided into 12, with eight repetitions with seven hens. These treatments were then distributed in a factorial design of 2 x 3 x 2 (two methodologies for beak trimming during rearing phase x three strains x with or without beak trimming at ten weeks of age). In the rearing phase the birds treated by infrared radiation had a lower total consumption and a longer beak length. In the growing phase, the execution of beak trimming at ten weeks of age, led to a lower total consumption and a lower body weight, but a better food conversion, when compared to those hens that did not have beak treatment. Beak trimming at ten weeks did not change the plasmatic levels of corticosterone.

Production indicators and levels of corticosterone in pullets treated with beak-trimming protocols

The aim of this study was to compare the effects of beak trimming by hot blade or infrared radiation on production indicators and plasma levels of corticosterone in pullets. During the rearing phase, an entirely random delineation was used a 2 x 3 factorial design (two different techniques for beak trimming x 3 strains), a total of six treatments with six repetitions of 20 hens. In the growing phase, the previous treatments were divided into 12, with eight repetitions with seven hens. These treatments were then distributed in a factorial design of 2 x 3 x 2 (two methodologies for beak trimming during rearing phase x three strains x with or without beak trimming at ten weeks of age). In the rearing phase the birds treated by infrared radiation had a lower total consumption and a longer beak length. In the growing phase, the execution of beak trimming at ten weeks of age, led to a lower total consumption and a lower body weight, but a better food conversion, when compared to those hens that did not have beak treatment. Beak trimming at ten weeks did not change the plasmatic levels of corticosterone.(AU)

Conforto térmico de ovinos da raça santa inês confinados com dietas contendo três níveis de inclusão de concentrado / Thermal comfort of santa ines sheep fed diets containing three levels of inclusion of concentrates

Objetivou-se avaliar o conforto térmico de ovinos confinados recebendo dietas com a inclusão de três níveis de concentrado. Foram utilizados 15 animais machos, da raça Santa Inês, peso médio 17±3,32 kg e idades entre 4 meses e 1 ano, recebendo três níveis de inclusão de concentrado (20, 40 e 60%). Foram determinadas as variáveis ambientais (temperatura do ar (TA), temperatura máxima (Tmáx) e mínima (Tmín), umidade relativa do ar (UR) e velocidade do vento (VV)) e os parâmetros fisiológicos (frequência respiratória (FR), temperatura retal (TR) e superficial (TS)) às 8:00, 12:00, 14:00 e 17:00 h. O delineamento utilizado foi em blocos casualizados arranjados em parcelas subdivididas, utilizando análise de regressão para TA, Tmáx, Tmín, UR e VV, contrastes ortogonais para desdobramentos da interações e teste Tukey para análise das médias das variáveis FR, TR e TS. A inclusão de concentrado na dieta de ovinos acima de 20% afetou o conforto térmico dos animais nas condições ambientais apresentadas.(AU)

This study aimed to evaluate the thermal comfort of feedlot sheep fed diet with the inclusion of three different concentrate levels. 15 males from Santa Ines breed were used, average weight 17±3.32 kg, age from 4 months to 1 year, receiving three levels of concentrate (20, 40 and 60%). The environmental variables were determined such as air temperature (AT), maximum temperature (maxT) and minimum (minT), relative humidity (RH) and wind speed (WS)) and physiological parameters (respiratory rate (RR), rectal temperature (RT) and surface (ST) at 8:00 a.m., 12:00 p.m., 2:00 p.m. and 5:00 p.m..The design was randomized blocks arranged in a split plot, using regression analysis for AT, maxT, minT, RH and WS, orthogonal contrasts for unfolding interactions and Tukey test for the analysis of the means of the variables RR, RT and ST. The inclusion of concentrate in the diet of sheep over 20% affects thermal comfort.(AU)

Conforto térmico de ovinos da raça santa inês confinados com dietas contendo três níveis de inclusão de concentrado / Thermal comfort of santa ines sheep fed diets containing three levels of inclusion of concentrates

Objetivou-se avaliar o conforto térmico de ovinos confinados recebendo dietas com a inclusão de três níveis de concentrado. Foram utilizados 15 animais machos, da raça Santa Inês, peso médio 17±3,32 kg e idades entre 4 meses e 1 ano, recebendo três níveis de inclusão de concentrado (20, 40 e 60%). Foram determinadas as variáveis ambientais (temperatura do ar (TA), temperatura máxima (Tmáx) e mínima (Tmín), umidade relativa do ar (UR) e velocidade do vento (VV)) e os parâmetros fisiológicos (frequência respiratória (FR), temperatura retal (TR) e superficial (TS)) às 8:00, 12:00, 14:00 e 17:00 h. O delineamento utilizado foi em blocos casualizados arranjados em parcelas subdivididas, utilizando análise de regressão para TA, Tmáx, Tmín, UR e VV, contrastes ortogonais para desdobramentos da interações e teste Tukey para análise das médias das variáveis FR, TR e TS. A inclusão de concentrado na dieta de ovinos acima de 20% afetou o conforto térmico dos animais nas condições ambientais apresentadas.

This study aimed to evaluate the thermal comfort of feedlot sheep fed diet with the inclusion of three different concentrate levels. 15 males from Santa Ines breed were used, average weight 17±3.32 kg, age from 4 months to 1 year, receiving three levels of concentrate (20, 40 and 60%). The environmental variables were determined such as air temperature (AT), maximum temperature (maxT) and minimum (minT), relative humidity (RH) and wind speed (WS)) and physiological parameters (respiratory rate (RR), rectal temperature (RT) and surface (ST) at 8:00 a.m., 12:00 p.m., 2:00 p.m. and 5:00 p.m..The design was randomized blocks arranged in a split plot, using regression analysis for AT, maxT, minT, RH and WS, orthogonal contrasts for unfolding interactions and Tukey test for the analysis of the means of the variables RR, RT and ST. The inclusion of concentrate in the diet of sheep over 20% affects thermal comfort.

Osteogenic differentiation of adipose tissue-derived mesenchymal stem cells cultured with different concentrations of prolactin / Diferenciação osteogênica de células tronco mesenquimais do tecido adiposo cultivadas com diferentes concentrações de prolactina

The objective was to evaluate the in vitro effect of prolactin in osteogenic potential of adipose tissue-derived mesenchymal stem cells (ADSCs) in female rats. ADSCs were cultured in osteogenic medium with and without the addition of prolactin and distributed into three groups: 1) ADSCs (control), 2) ADSCs with addition of 100ng/mL of prolactin and 3) ADSCs with addition of 300ng/mL of prolactin. At 21 days of differentiation, the tests of MTT conversion into formazan crystals, percentage of mineralized nodules and cells per field and quantification of genic transcript for alkaline phosphatase, osteopontin, osteocalcin, bone sialoprotein, BMP-2 and collagen I by real-time RT-PCR were made. The addition of prolactin reduced the conversion of MTT in group 3 and increased the percentage of cells per field in the groups 2 and 3, however without significantly increasing the percentage of mineralized nodules and the expression of alkaline phosphatase, osteopontin, osteocalcin, bone sialoprotein, BMP-2 and collagen I. In conclusion, the addition of prolactin in concentrations of 100ng/mL and 300ng/mL does not change the osteogenic differentiation to the ADSCs of female rats despite increase in the cellularity of the culture.(AU)

O objetivo do presente trabalho foi avaliar o efeito in vitro da prolactina sobre o potencial osteogênico de células-tronco mesenquimais do tecido adiposo (CTM-TA) em ratas. CTM-TA foram cultivadas em meio osteogênico com e sem adição de prolactina e distribuídas em três grupos: 1) CTM-TA (controle), 2) CM-TA com adição de 100ng/mL de prolactina e 3) CTM-TA com adição de 300ng/mL de prolactina. Aos 21 dias de diferenciação, foram realizados os testes de conversão do MTT em cristais de formazan, porcentagem de nódulos mineralizados e células por campo e quantificação dos transcritos gênicos para fosfatase alcalina, osteopontina, osteocalcina, sialoproteína óssea, BMP-2 e colágeno I. A adição de prolactina reduziu a conversão do MTT no grupo 3 e aumentou a porcentagem de células por campo nos grupos 2 e 3, sem alterar significativamente a porcentagem de nódulos mineralizados e a expressão de fosfatase alcalina, osteopontina, osteocalcina, sialoproteína óssea, BMP-2 e colágeno I. Conclui-se que a adição de prolactina nas concentrações de 100ng/mL e 300ng/mL não altera a diferenciação osteogênica das CTM-TA de ratas, apesar do aumento de celularidade da cultura.(AU)

Osteogenic differentiation of adipose tissue-derived mesenchymal stem cells cultured with different concentrations of prolactin / Diferenciação osteogênica de células tronco mesenquimais do tecido adiposo cultivadas com diferentes concentrações de prolactina

The objective was to evaluate the in vitro effect of prolactin in osteogenic potential of adipose tissue-derived mesenchymal stem cells (ADSCs) in female rats. ADSCs were cultured in osteogenic medium with and without the addition of prolactin and distributed into three groups: 1) ADSCs (control), 2) ADSCs with addition of 100ng/mL of prolactin and 3) ADSCs with addition of 300ng/mL of prolactin. At 21 days of differentiation, the tests of MTT conversion into formazan crystals, percentage of mineralized nodules and cells per field and quantification of genic transcript for alkaline phosphatase, osteopontin, osteocalcin, bone sialoprotein, BMP-2 and collagen I by real-time RT-PCR were made. The addition of prolactin reduced the conversion of MTT in group 3 and increased the percentage of cells per field in the groups 2 and 3, however without significantly increasing the percentage of mineralized nodules and the expression of alkaline phosphatase, osteopontin, osteocalcin, bone sialoprotein, BMP-2 and collagen I. In conclusion, the addition of prolactin in concentrations of 100ng/mL and 300ng/mL does not change the osteogenic differentiation to the ADSCs of female rats despite increase in the cellularity of the culture.(AU)

O objetivo do presente trabalho foi avaliar o efeito in vitro da prolactina sobre o potencial osteogênico de células-tronco mesenquimais do tecido adiposo (CTM-TA) em ratas. CTM-TA foram cultivadas em meio osteogênico com e sem adição de prolactina e distribuídas em três grupos: 1) CTM-TA (controle), 2) CM-TA com adição de 100ng/mL de prolactina e 3) CTM-TA com adição de 300ng/mL de prolactina. Aos 21 dias de diferenciação, foram realizados os testes de conversão do MTT em cristais de formazan, porcentagem de nódulos mineralizados e células por campo e quantificação dos transcritos gênicos para fosfatase alcalina, osteopontina, osteocalcina, sialoproteína óssea, BMP-2 e colágeno I. A adição de prolactina reduziu a conversão do MTT no grupo 3 e aumentou a porcentagem de células por campo nos grupos 2 e 3, sem alterar significativamente a porcentagem de nódulos mineralizados e a expressão de fosfatase alcalina, osteopontina, osteocalcina, sialoproteína óssea, BMP-2 e colágeno I. Conclui-se que a adição de prolactina nas concentrações de 100ng/mL e 300ng/mL não altera a diferenciação osteogênica das CTM-TA de ratas, apesar do aumento de celularidade da cultura.(AU)

Testicular Structure and Seminal Pathway in the Yellowtail Tetra Astyanax altiparanae (Characiforms: Characidae).

This study aimed to describe testicular and its main ducts structure in the yellowtail tetra Astyanax altiparanae, contributing to the knowledge of the region in which semen is produced, storage and released, focusing mainly on the dynamic of germinal epithelium and Sertoli cells during germ cell maturation. Ten sexually mature male A. altiparanae had their testes processed according to the routine protocols to optical microscopy. Moreover, spermatic ducts and tubular compartment of the testes of three specimens were perfused with vinyl resin for gross anatomy and scanning electron microscopy. Astyanax altiparanae testes are paired organs, separated for most of their extension, joining posteriorly in a spermatic duct formed by a squamous simple epithelium. Seminiferous compartment presents anastomosing tubular type organisation, and spermatogonia spread along its extent. Spermatogenesis is of cystic type, and there is no main testicular duct. Spermatogenesis develops in 'waves', from posterior to anterior part of the gonad. Thus, while sperm is storage posteriorly, spermatogenesis keeps maturing germ cells anteriorly, making the germinal epithelium very dynamic, holding Sertoli cells that change their function as a cystic envelope to produce secretions of the seminal fluid and store sperm. Such kind of development is thought to be responsible by the high prolificacy of this species.

Osteogenic differentiation of adipose tissue-derived mesenchymal stem cells cultured with different concentrations of prolactin

ABSTRACT The objective was to evaluate the in vitro effect of prolactin in osteogenic potential of adipose tissue-derived mesenchymal stem cells (ADSCs) in female rats. ADSCs were cultured in osteogenic medium with and without the addition of prolactin and distributed into three groups: 1) ADSCs (control), 2) ADSCs with addition of 100ng/mL of prolactin and 3) ADSCs with addition of 300ng/mL of prolactin. At 21 days of differentiation, the tests of MTT conversion into formazan crystals, percentage of mineralized nodules and cells per field and quantification of genic transcript for alkaline phosphatase, osteopontin, osteocalcin, bone sialoprotein, BMP-2 and collagen I by real-time RT-PCR were made. The addition of prolactin reduced the conversion of MTT in group 3 and increased the percentage of cells per field in the groups 2 and 3, however without significantly increasing the percentage of mineralized nodules and the expression of alkaline phosphatase, osteopontin, osteocalcin, bone sialoprotein, BMP-2 and collagen I. In conclusion, the addition of prolactin in concentrations of 100ng/mL and 300ng/mL does not change the osteogenic differentiation to the ADSCs of female rats despite increase in the cellularity of the culture.

RESUMO O objetivo do presente trabalho foi avaliar o efeito in vitro da prolactina sobre o potencial osteogênico de células-tronco mesenquimais do tecido adiposo (CTM-TA) em ratas. CTM-TA foram cultivadas em meio osteogênico com e sem adição de prolactina e distribuídas em três grupos: 1) CTM-TA (controle), 2) CM-TA com adição de 100ng/mL de prolactina e 3) CTM-TA com adição de 300ng/mL de prolactina. Aos 21 dias de diferenciação, foram realizados os testes de conversão do MTT em cristais de formazan, porcentagem de nódulos mineralizados e células por campo e quantificação dos transcritos gênicos para fosfatase alcalina, osteopontina, osteocalcina, sialoproteína óssea, BMP-2 e colágeno I. A adição de prolactina reduziu a conversão do MTT no grupo 3 e aumentou a porcentagem de células por campo nos grupos 2 e 3, sem alterar significativamente a porcentagem de nódulos mineralizados e a expressão de fosfatase alcalina, osteopontina, osteocalcina, sialoproteína óssea, BMP-2 e colágeno I. Conclui-se que a adição de prolactina nas concentrações de 100ng/mL e 300ng/mL não altera a diferenciação osteogênica das CTM-TA de ratas, apesar do aumento de celularidade da cultura.

Evaluation of Recurrence Predictors and Survival Probability After Liver Transplantation for Hepatocellular Carcinoma: Analysis From a Single Center.

BACKGROUND: Liver transplantation is the criterion standard treatment for hepatocellular carcinoma, but tumor recurrence remains a problem leading to a worse prognosis. We aimed to evaluate factors associated with recurrence of hepatocellular carcinoma and survival after this procedure. METHODS: We retrospectively reviewed data from 101 patients submitted to liver transplantation because of hepatocellular carcinoma from January 2005 to January 2012 at our single center. Univariate and multivariate analyses were performed to identify preoperative factors and histologic findings associated with lower survival rates and recurrence. The significance level was 5%. RESULTS: There was recurrence in 10 cases (9.9%), with an average time of 25.28 ± 26.92 months. Microvascular invasion (P = .005; hazard ratio [HR], 4.94; 95% confidence interval [CI], 1.42-17.12) was an independent factor for recurrence. Microvascular invasion (P = .035; HR, 1.87; 95% CI, 1.04-3.25) and tumors outside the criteria of the University of San Francisco (P = .046; HR, 1.81; 95% CI, 1.01-3.25) were independent factors for the risk of death. Poorly differentiated tumors had a higher level of alphafetoprotein (P = .03), and values <100 ng/mL were associated with well differentiated tumors. CONCLUSIONS: Microvascular invasion was associated with recurrence and lower survival. Tumors outside the criteria of the University of California, San Francisco had lower survival. Alpha-fetoprotein levels >100 ng/mL were associated with poorly differentiated tumors.

Can microcystins affect zooplankton structure community in tropical eutrophic reservoirs? / Microcistinas podem afetar a estrutura da comunidade zooplanctônica em reservatórios eutróficos tropicais?

Abstract The aim of our study was to assess whether cyanotoxins (microcystins) can affect the composition of the zooplankton community, leading to domination of microzooplankton forms (protozoans and rotifers). Temporal variations in concentrations of microcystins and zooplankton biomass were analyzed in three eutrophic reservoirs in the semi-arid northeast region of Brazil. The concentration of microcystins in water proved to be correlated with the cyanobacterial biovolume, indicating the contributions from colonial forms such as Microcystis in the production of cyanotoxins. At the community level, the total biomass of zooplankton was not correlated with the concentration of microcystin (r2 = 0.00; P > 0.001), but in a population-level analysis, the biomass of rotifers and cladocerans showed a weak positive correlation. Cyclopoid copepods, which are considered to be relatively inefficient in ingesting cyanobacteria, were negatively correlated (r2 = 0.01; P > 0.01) with the concentration of cyanotoxins. Surprisingly, the biomass of calanoid copepods was positively correlated with the microcystin concentration (r2 = 0.44; P > 0.001). The results indicate that allelopathic control mechanisms (negative effects of microcystin on zooplankton biomass) do not seem to substantially affect the composition of mesozooplankton, which showed a constant and high biomass compared to the microzooplankton (rotifers). These results may be important to better understand the trophic interactions between zooplankton and cyanobacteria and the potential effects of allelopathic compounds on zooplankton.(AU)

Resumo Com o objetivo de avaliar se as cianotoxinas (microcistinas) podem afetar a composição da comunidade zooplanctônica, levando à dominância de formas microzooplanctônicas (protozoários e rotiferos), as variações nas concentrações de microcistina e a biomassa do zooplâncton foram analisadas em três reservatórios eutróficos na região semi-árida do nordeste brasileiro. A concentração de microcistinas na água esteve correlacionada com o biovolume de cianobactérias, indicando a contribuição de formas coloniais como Microcystis na produção de cianotoxinas. A nível de comunidade, a biomassa total do zooplâncton não apresentou correlacão com a concentração de microcistina (r2 = 0.00; P > 0.001), mas em uma análise a nível de populações, a biomassa de rotíferos e cladóceros apresentou uma fraca correlação positiva. Copépodos Cyclopoida, os quais são considerados relativamente ineficientes na ingestão de cianobactérias, estiveram negativamente correlacionados com a concentração de microcistinas (r2 = - 0.01; P > 0.01). Surpreendentemente, a biomassa de copépodos Calanoida foi positivamente correlacionada com a concentração de cianotoxinas (r2 = 0.44; P > 0.001). Os resultados indicam que mecanismos de controle alelopáticos (efeitos negativos da microcistina sobre o zooplâncton) parecem não afetar substancialmente a composição do mesozooplâncton, que apresentou uma alta e constante biomassa, quando comparada à biomassa do microzooplâncton (rotíferos). Esses resultados podem ser importantes para um melhor entendimento das interações tróficas entre o zooplâncton e cianobactérias, e do efeito potencial de compostos alelopáticos sobre o zooplâncton.(AU)