Gestational protein restriction induces CA3 dendritic atrophy in dorsal hippocampal neurons but does not alter learning and memory performance in adult offspring

Studies have demonstrated that nutrient deficiency during pregnancy or in early postnatal life results in structural abnormalities in the offspring hippocampus and in cognitive impairment. In an attempt to analyze whether gestational protein restriction might induce learning and memory impairments associated with structural changes in the hippocampus, we carried out a detailed morphometric analysis of the hippocampus of male adult rats together with the behavioral characterization of these animals in the Morris water maze (MWM). Our results demonstrate that gestational protein restriction leads to a decrease in total basal dendritic length and in the number of intersections of CA3 pyramidal neurons whereas the cytoarchitecture of CA1 and dentate gyrus remained unchanged. Despite presenting significant structural rearrangements, we did not observe impairments in the MWM test. Considering the clear dissociation between the behavioral profile and the hippocampus neuronal changes, the functional significance of dendritic remodeling in fetal processing remains undisclosed. © 2012 ISDN.

Major components of energy drinks (caffeine, taurine, and guarana) exert cytotoxic effects on human neuronal SH-SY5Y cells by decreasing reactive oxygen species production

Scope. To elucidate the morphological and biochemical in vitro effects exerted by caffeine, taurine, and guarana, alone or in combination, since they are major components in energy drinks (EDs). Methods and Results. On human neuronal SH-SY5Y cells, caffeine (0.125-2 mg/mL), taurine (1-16 mg/mL), and guarana (3.125-50 mg/mL) showed concentration-dependent nonenzymatic antioxidant potential, decreased the basal levels of free radical generation, and reduced both superoxide dismutase (SOD) and catalase (CAT) activities, especially when combined together. However, guarana-treated cells developed signs of neurite degeneration in the form of swellings at various segments in a beaded or pearl chain-like appearance and fragmentation of such neurites at concentrations ranging from 12.5 to 50 mg/mL. Swellings, but not neuritic fragmentation, were detected when cells were treated with 0.5 mg/mL (or higher doses) of caffeine, concentrations that are present in EDs. Cells treated with guarana also showed qualitative signs of apoptosis, including membrane blebbing, cell shrinkage, and cleaved caspase-3 positivity. Flow cytometric analysis confirmed that cells treated with 12.5-50 mg/mL of guarana and its combinations with caffeine and/or taurine underwent apoptosis. Conclusion. Excessive removal of intracellular reactive oxygen species, to nonphysiological levels (or antioxidative stress), could be a cause of in vitro toxicity induced by these drugs. © 2013 Fares Zeidán-Chuliá et al.

Bone morphogenetic protein 15 and fibroblast growth factor 10 enhance cumulus expansion, glucose uptake, and expression of genes in the ovulatory cascade during in vitro maturation of bovine cumulus-oocyte complexes

Oocyte-secreted factors (OSFs) regulate differentiation of cumulus cells and are of pivotal relevance for fertility. Bone morphogenetic protein 15 (BMP15) and fibroblast growth factor 10 (FGF10) are OSFs and enhance oocyte competence by unknown mechanisms. We tested the hypothesis that BMP15 and FGF10, alone or combined in the maturation medium, enhance cumulus expansion and expression of genes in the preovulatory cascade and regulate glucose metabolism favouring hyaluronic acid production in bovine cumulus-oocyte complexes (COCs). BMP15 or FGF10 increased the percentage of fully expanded COCs, but the combination did not further stimulate it. BMP15 increased cumulus cell levels of mRNA encoding a disintegrin and metalloprotease 10 (ADAM10), ADAM17, amphiregulin (AREG), and epiregulin (EREG) at 12 h of culture and of prostaglandin (PG)-endoperoxide synthase 2 (PTGS2), pentraxin 3 (PTX3) and tumor necrosis factor alpha-induced protein 6 (TNFAIP6 (TSG6)) at 22 h of culture. FGF10 did not alter the expression of epidermal growth factor-like factors but enhanced the mRNA expression of PTGS2 at 4 h, PTX3 at 12 h, and TNFAIP6 at 22 h. FGF10 and BMP15 stimulated glucose consumption by cumulus cells but did not affect lactate production or levels of mRNA encoding glycolytic enzymes phosphofructokinase and lactate dehydrogenase A. Each growth factor increased mRNA encoding glucosamine:fructose-6-PO4 transaminases, key enzymes in the hexosamine pathway leading to hyaluronic acid production, and BMP15 also stimulated hyaluronan synthase 2 (HAS2) mRNA expression. This study provides evidence that BMP15 and FGF10 stimulate expansion of in vitro-matured bovine COCs by driving glucose metabolism toward hyaluronic acid production and controlling the expression of genes in the ovulatory cascade, the first acting upon ADAM10, ADAM17, AREG, and EREG and the second on downstream genes, particularly PTGS2. © 2013 Society for Reproduction and Fertility.

Acute phase protein profile in agoutis (Dasyprocta azarae; Lichtenstein, 1823) in captivity, determined by sodium dodecyl sulfate polyacrilamyde gel electrophoresis

The acute phase response refers to a nonspecific and complex systemic reaction of the organism that occurs shortly after any tissue injury. The acute phase response is considered a part of the innate host defense system, which is responsible for the survival of the host during the critical early stages of attack, and in evolutionary terms, it precedes the acquired immune response. The purpose of this study was to determine serum protein concentrations, including the acute phase protein profile in agoutis (Dasyprocta azarae) in captivity, by means of sodium dodecyl sulfate polyacrylamide gel electrophoresis. Blood samples from 11 adult healthy animals (nine females and two males) were obtained. The serum proteinogram had 21 proteins with molecular weights ranging from 15 to 240 kD. The acute phase proteins identified were: ceruloplasmin, transferrin, albumin, haptoglobin, α-1-acid glycoprotein, and hemoglobin. IgA, IgG heavy and light chains, and nonnominal identified proteins of 240, 210, 140, 98, 78, 48, 35, 31, 23, and 15 kD were also identified. The determination of the acute phase protein concentrations is a useful method for the early detection of subclinical disease or changes in the healthy animal, with predictive information on the development of disease in the future. It is possible to standardize the reference values of the serum protein profile of agoutis, which can be used for diagnosis and prognosis, treatment and clinical follow-up of nutritional disorders, and immune-mediated inflammatory diseases that may affect these animals. © 2012 Springer-Verlag London Limited.

STEAP1 protein overexpression is an independent marker for biochemical recurrence in prostate carcinoma

Aims: To investigate the prognostic value of expression levels of the genes STEAP1 and STEAP2, and of STEAP1 protein, in prostate carcinomas (PCa). Methods and results: STEAP1 and STEAP2 transcript levels were evaluated by RT-qPCR in samples from 35 PCa, 24 adjacent non-neoplastic prostate (AdjP) tissues, five cases of benign prostatic hyperplasia (BPH), and two histologically normal prostates (N). STEAP1 expression was assessed by immunohistochemistry in samples from 198 PCa, 76 AdjP, 22 BPH, and two N. The findings were compared with clinical and pathological parameters and patient outcome. STEAP1 and STEAP2 transcript analysis showed no differences between the groups tested. Although not significant, higher STEAP1 mRNA levels were detected in tumours with high Gleason scores and in patients who presented with biochemical recurrence (BCR). STEAP1 overexpression was detected in PCa, and was significantly associated with high-grade Gleason scores, seminal vesicle invasion, BCR, and worse outcome (metastasis or PCa-specific death). STEAP1 overexpression was significantly associated with shorter BCR-free survival. Multivariate analysis revealed that STEAP1 is an independent marker for BCR. Conclusions: These findings provide evidence that STEAP1 is a biomarker of worse prognosis in PCa patients. © 2013 John Wiley & Sons Ltd.

Triiodothyronine Increases mRNA and Protein Leptin Levels in Short Time in 3T3-L1 Adipocytes by PI3K Pathway Activation

The present study aimed to examine the effects of thyroid hormone (TH), more precisely triiodothyronine (T3), on the modulation of leptin mRNA expression and the involvement of the phosphatidyl inositol 3 kinase (PI3K) signaling pathway in adipocytes, 3T3-L1, cell culture. We examined the involvement of this pathway in mediating TH effects by treating 3T3-L1 adipocytes with physiological (P=10nM) or supraphysiological (SI=100 nM) T3 dose during one hour (short time), in the absence or the presence of PI3K inhibitor (LY294002). The absence of any treatment was considered the control group (C). RT-qPCR was used for mRNA expression analyzes. For data analyzes ANOVA complemented with Tukey's test was used at 5% significance. T3 increased leptin mRNA expression in P (2.26 ± 0.36, p< 0.001), SI (1.99 ±0.22, p< 0.01) compared to C group (1± 0.18). This increase was completely abrogated by LY294002 in P (1.31±0.05, p< 0.001) and SI (1.33±0.31, p< 0.05). Western blotting confirmed these results at protein level, indicating the PI3K pathway dependency. To examine whether leptin is directly induced by T3, we used the translation inhibitor cycloheximide (CHX). In P, the presence of CHX maintained the levels mRNA leptin, but was completely abrogated in SI (1.14±0.09, p> 0.001). These results demonstrate that the activation of the PI3K signaling pathway has a role in TH-mediated direct and indirect leptin gene expression in 3T3-L1 adipocytes. © 2013 Oliveira et al.

Digestible tryptophan requirements for broilers from 22 to 42 days old

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Lesão do nervo fibular comum. Avaliação neuronal, muscular e funcional após neurotmese parcial: estudo no rato

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Critérios de avaliação das exigências em treonina, triptofano, valina e isoleucina para frangos de corte de 22 a 42 dias de idade

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Expression of P-glycoprotein, multidrug resistance-associated protein, glutathione-S-transferase pi and p53 in canine transmissible venereal tumor

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Digestible valine requirements for broilers from 22 and 42 days old

Current experiment established different criteria to evaluate the requirements of digestible valine for broilers from 22 and 42 days of age, by different regression models (quadratic, exponential and Linear Response Plateau) and, in the case of statistical significance, the comparison of means by Duncan test at 5% probability. A total of 1,920 Cobb 500 male broilers were used and distributed in an entirely randomized experimental design, with 6 treatments (6 digestible valine levels: 0.7192, 0.7729, 0.8265, 0.8802, 0.9339 and 0.9876%) and 8 replications, with 40 broilers each. Data on performance and carcass characteristics were evaluated. The level of 0.8265% digestible valine was considered standard. The inclusion of 0.816, 0.848 and 0.903% of digestible valine levels, corresponding to digestible valine:lysine ratios of approximately 76.00%, 79.00% and 84.12%, provided best feed intake, weight gain and feed conversion ratio, respectively for broiler from 22 to 42 days of age.

Supplementation based on protein or energy ingredients to beef cattle consuming low-quality cool-season forages: I. Forage disappearance parameters in rumen-fistulated steers and physiological responses in pregnant heifers

Two experiments evaluated the influence of supplement composition on ruminal forage disappearance, performance, and physiological responses of Angus x Hereford cattle consuming a low-quality cool-season forage (8.7% CP and 57% TDN). In Exp. 1, 6 rumen-fistulated steers housed in individual pens were assigned to an incomplete 3 x 2 Latin square design containing 2 periods of 11 d each and the following treatments: 1) supplementation with soybean meal (PROT), 2) supplementation with a mixture of cracked corn, soybean meal, and urea (68:22:10 ratio, DM basis; ENER), or 3) no supplementation (CON). Steers were offered meadow foxtail (Alopecurus pratensis L.) hay for ad libitum consumption. Treatments were provided daily at 0.50 and 0.54% of shrunk BW/steer for PROT and ENER, respectively, to ensure that PROT and ENER intakes were isocaloric and isonitrogenous. No treatment effects were detected on rumen disappearance parameters of forage DM (P >= 0.33) and NDF (P >= 0.66). In Exp. 2, 35 pregnant heifers were ranked by initial BW on d -7 of the study, allocated into 12 feedlot pens (4 pens/treatment), and assigned to the same treatments and forage intake regimen as in Exp. 1 for 19 d. Treatments were fed once daily at 1.77 and 1.92 kg of DM/heifer for PROT and ENER, respectively, to achieve the same treatment intake as percent of initial BW used in Exp. 1 (0.50 and 0.54% for PROT and ENER, respectively). No treatment effects (P = 0.17) were detected on forage DMI. Total DMI was greater (P < 0.01) for PROT and ENER compared with CON and similar between PROT and ENER (P = 0.36). Accordingly, ADG was greater (P = 0.01) for PROT compared with CON, tended to be greater for ENER compared with CON (P = 0.08), and was similar between ENER and PROT (P = 0.28). Heifers receiving PROT and ENER had greater mean concentrations of plasma glucose (P = 0.03), insulin (P <= 0.09), IGF-I (P <= 0.04), and progesterone (P = 0.01) compared to CON, whereas ENER and PROT had similar concentrations of these variables (P >= 0.15). A treatment x hour interaction was detected (P < 0.01) for plasma urea N (PUN), given that PUN concentrations increased after supplementation for ENER and PROT (time effect, P < 0.01) but did not change for CON (time effect, P = 0.62). In conclusion, beef cattle consuming low-quality cool-season forages had similar ruminal forage disappearance and intake, performance, and physiological status if offered supplements based on soybean meal or corn at 0.5% of BW.

Circadian-rhythm of the endopeptidase 22.19 (ec 3.4.22.19) in the rat-brain

The endopeptidase 22.19 (EC 3.4.22.19) has been associated with the metabolism of neuropeptides by its ability to convert small enkephalin-containing peptides (8 to 13 amino acids) into enkephalins. In addition, this enzyme cleaves the Arg8-Arg9 bond of neurotensin and the Phe5-Ser6 bond of bradykinin. We analyzed the circadian variation of endopeptidase 22.19 in the whole and individual areas of the rat brain. Endopeptidase 22.19 activity was analyzed by high-performance liquid chromatography (HPLC) using bradykinin as an operative substrate. Enzymatic specific activities were analyzed by rhythmometric methods and indicate a circadian fluctuation of endopeptidase 22.19 specific activity (mU of enzyme/mg of protein) in the whole brain [p < 0.001, mesor (M) = 7.62, amplitude (A) = 2.89, and acrophase (phi) = 23:08 h], striatum (p < 0.001, M = 2.92, A = 0.62, phi-23:03 h), hypothalamus (p < 0.001, M = 3.15, A = 0.86, phi-01:12 h), periaqueductal gray matter (p < 0.005, M = 2.62, A = 0.34, phi = 22:35 h), and cerebellum (p < 0.0 14, M = 4.27, A = 0.88, phi = 17:12 h). The circadian rhythmicity in endopeptidase 22.19 specific activity suggests that light may have an effect on the peptidase activity in whole brain and in areas of the central nervous system and may be essential for the mechanisms of circadian fluctuations of neuropeptides in the brain.

Metabolism and ruminal parameters of Holstein × Gir heifers fed sugarcane and increasing levels of crude protein

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)