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.

Bone marrow mesenchymal stem cells stimulated by bFGF up-regulated protein expression in comparison with periodontal fibroblasts in vitro

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

Hybrid sorubim viscera protein concentrate in the diets for barred sorubim

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Hypusine Modification of the Ribosome-binding Protein eIF5A, a Target for New Anti-Inflammatory Drugs: Understanding the Action of the Inhibitor GC7 on a Murine Macrophage Cell Line

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

The effect of lipid sources on intake, rumen fermentation parameters and microbial protein synthesis in Nellore steers supplemented with glycerol

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

BtoxDB: a comprehensive database of protein structural data on toxin-antitoxin systems

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

Different true-protein sources do not modify the metabolism of crossbred Bos taurus x Bos indicus growing heifers

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

Effects of protein supplementation frequency on physiological responses associated with reproduction in beef cows

The objective of this experiment was to determine if frequency of protein supplementation impacts physiological responses associated with reproduction in beef cows. Fourteen nonpregnant, nonlactating beef cows were ranked by age and BW and allocated to 3 groups. Groups were assigned to a 3 x 3 Latin square design, containing 3 periods of 21 d and the following treatments: 1) soybean meal supplementation daily (D), 2) soybean meal supplementation 3 times/week (3WK), and 3) soybean meal supplementation once/week (1WK). Within each period, cows were assigned to an estrus synchronization protocol: 100 mu g of GnRH + controlled internal drug release device (CIDR) containing 1.38 g of progesterone (P-4) on d 1, 25 mg of PGF(2 alpha) on d 8, and CIDR removal + 100 mu g of GnRH on d 11. Grass-seed straw was offered for ad libitum consumption. Soybean meal was individually supplemented at a daily rate of 1 kg/cow (as-fed basis). Moreover, 3WK was supplemented on d 0, 2, 4, 7, 9, 11, 14, 16, and 18 whereas 1WK was supplemented on d 4, 11, and 18. Blood samples were collected from 0 (before) to 72 h after supplementation on d 11 and 18 and analyzed for plasma urea-N (PUN). Samples collected from 0 to 12 h were also analyzed for plasma glucose, insulin, and P-4 (d 18 only). Uterine flushing fluid was collected concurrently with blood sampling at 28 h for pH evaluation. Liver biopsies were performed concurrently with blood sampling at 0, 4, and 28 h and analyzed for mRNA expression of carbamoyl phosphate synthetase I (CPS-I; h 28) and CYP2C19 and CYP3A4 (h 0 and 4 on d 18). Plasma urea-N concentrations were greater (P < 0.01) for 1WK vs. 3WK from 20 to 72 h and greater (P < 0.01) for 1WK vs. D from 16 to 48 h and at 72 h after supplementation (treatment x hour interaction, P < 0.01). Moreover, PUN concentrations peaked at 28 h after supplementation for 3WK and 1WK (P < 0.01) and were greater (P < 0.01) at this time for 1WK vs. 3WK and D and for 3WK vs. D. Expression of CPS-I was greater (P < 0.01) for 1WK vs. D and 3WK. Uterine flushing pH tended (P <= 0.10) to be greater for 1WK vs. 3WK and D. No treatment effects were detected (P >= 0.15) on expression of CYP2C19 and CYP3A4, plasma glucose, and P-4 concentrations, whereas plasma insulin concentrations were greater (P <= 0.03) in D and 3WK vs. 1WK. Hence, decreasing frequency of protein supplementation did not reduce uterine flushing pH or plasma P-4 concentrations, which are known to impact reproduction in beef cows.

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)

Serum protein profile of hookworm infection in dogs

Hookworms are parasitic nematodes that cause anemia and intestinal infections in dogs, especially with large worm burdens. However, the serum protein profile of this parasitological disease is still poorly understood. The present study was design to evaluate 80 asymptomatic dogs (age; 8 months–2 years) to detect the presence of the hookworm thin-shelled, morulated eggs in faeces using faecal flotation and to evaluate the serum protein fractions determined by means of sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Forty dogs had an elevated quantity of eggs in faeces (+++) (PD) and 40 dogs were healthy (HD). Infected dogs showed significant increases for IgG estimated concentrations (PD 1.79±0.8 g/dL and HD 1.44±0.72 g/dL, p= 0.04), for ceruloplasmin estimated concentrations (PD 19±15 mg/dL and HD 5±3.5 mg/dL, p=0.0001), for alpha 1-acid glycoprotein estimated concentrations (PD 31.4±17.9 mg/dL and HD 13.5±12.1 mg/dL, p=0.0001) and for a non-identified protein of 23 kDa estimated concentrations (PD 641.5±194.9 mg/dL and HD 519.8± 197.9 mg/dL, p=0.007). Dogs with hookworm infection showed significant differences in the serum protein profile when compared to healthy animals.

Linker for Activation of T-cell Family Member2 (LAT2) a Lipid Raft Adaptor Protein for AKT Signaling, Is an Early Mediator of Alkylphospholipid Anti-leukemic Activity

Lipid rafts are highly ordered membrane domains rich in cholesterol and sphingolipids that provide a scaffold for signal transduction proteins; altered raft structure has also been implicated in cancer progression. We have shown that 25 mu M 10-(octyloxy) decyl-2-(trimethylammonium) ethyl phosphate (ODPC), an alkylphospholipid, targets high cholesterol domains in model membranes and induces apoptosis in leukemia cells but spares normal hematopoietic and epithelial cells under the same conditions. We performed a quantitative (SILAC) proteomic screening of ODPC targets in a lipid-raft-enriched fraction of leukemic cells to identify early events prior to the initiation of apoptosis. Six proteins, three with demonstrated palmitoylation sites, were reduced in abundance. One, the linker for activation of T-cell family member 2 (LAT2), is an adaptor protein associated with lipid rafts in its palmitoylated form and is specifically expressed in B lymphocytes and myeloid cells. Interestingly, LAT2 is not expressed in K562, a cell line more resistant to ODPC-induced apoptosis. There was an early loss of LAT2 in the lipid-raft-enriched fraction of NB4 cells within 3 h following treatment with 25 mu M ODPC. Subsequent degradation of LAT2 by proteasomes was observed. Twenty-five mu M ODPC inhibited AKT activation via myeloid growth factors, and LAT2 knockdown in NB4 cells by shRNA reproduced this effect. LAT2 knockdown in NB4 cells also decreased cell proliferation and increased cell sensitivity to ODPC (7.5X), perifosine (3X), and arsenic trioxide (8.5X). Taken together, these data indicate that LAT2 is an early mediator of the anti-leukemic activity of alkylphospholipids and arsenic trioxide. Thus, LAT2 may be used as a target for the design of drugs for cancer therapy. Molecular & Cellular Proteomics 11: 10.1074/mcp.M112.019661, 1898-1912, 2012.

Protein diets promote the maturation of oocytes and spawning of Piaractus mesopotamicus kept in cages

The objective of this study was to evaluate the effects of balanced diets on the maturation of oocytes and the reproductive performance of P. mesopotamicus in cages. A completely random design with 224 fish in 16 cages measuring 5 m(3) was employed for this purpose. The treatments consisted of diets containing 18, 24, 30, and 36% crude protein (CP) provided ad libitum. The external and internal morphological characteristics of the specimens were examined, as well as: the position of the germinal vesicle, the distribution of oocyte diameters, the fertilization and hatching rates, the number of oocytes released, the total number of oocytes, the remaining weight and total weight of the ovaries, the gonadosomatic index, the condition factor (K), and the histology of the oocytes and ovaries post-spawning and during ovarian regression. The diameters of the oocytes collected before the first hormonal application displayed a unimodal distribution for the lowest protein content and a polymodal distribution for the other treatments. A similar situation was seen during spawning. The lowest fertilization and hatching rates were found as a consequence of the treatment with 30% CP (P < 0.05). The greatest hatching rate occurred in the females fed 18% CP. The greatest total oocyte weight was found in the specimens that received between 30 and 36% CP. The lowest K index was found in the females fed 36% CP. In conclusion, a diet containing 18% CP satisfies the reproductive requirements of females adapted to this system.

Automatic design of decision-tree induction algorithms tailored to flexible-receptor docking data

Background: This paper addresses the prediction of the free energy of binding of a drug candidate with enzyme InhA associated with Mycobacterium tuberculosis. This problem is found within rational drug design, where interactions between drug candidates and target proteins are verified through molecular docking simulations. In this application, it is important not only to correctly predict the free energy of binding, but also to provide a comprehensible model that could be validated by a domain specialist. Decision-tree induction algorithms have been successfully used in drug-design related applications, specially considering that decision trees are simple to understand, interpret, and validate. There are several decision-tree induction algorithms available for general-use, but each one has a bias that makes it more suitable for a particular data distribution. In this article, we propose and investigate the automatic design of decision-tree induction algorithms tailored to particular drug-enzyme binding data sets. We investigate the performance of our new method for evaluating binding conformations of different drug candidates to InhA, and we analyze our findings with respect to decision tree accuracy, comprehensibility, and biological relevance. Results: The empirical analysis indicates that our method is capable of automatically generating decision-tree induction algorithms that significantly outperform the traditional C4.5 algorithm with respect to both accuracy and comprehensibility. In addition, we provide the biological interpretation of the rules generated by our approach, reinforcing the importance of comprehensible predictive models in this particular bioinformatics application. Conclusions: We conclude that automatically designing a decision-tree algorithm tailored to molecular docking data is a promising alternative for the prediction of the free energy from the binding of a drug candidate with a flexible-receptor.

Extensive structural change of the envelope protein of dengue virus induced by a tuned ionic strength: conformational and energetic analyses

The Dengue has become a global public health threat, with over 100 million infections annually; to date there is no specific vaccine or any antiviral drug. The structures of the envelope (E) proteins of the four known serotype of the dengue virus (DENV) are already known, but there are insufficient molecular details of their structural behavior in solution in the distinct environmental conditions in which the DENVs are submitted, from the digestive tract of the mosquito up to its replication inside the host cell. Such detailed knowledge becomes important because of the multifunctional character of the E protein: it mediates the early events in cell entry, via receptor endocytosis and, as a class II protein, participates determinately in the process of membrane fusion. The proposed infection mechanism asserts that once in the endosome, at low pH, the E homodimers dissociate and insert into the endosomal lipid membrane, after an extensive conformational change, mainly on the relative arrangement of its three domains. In this work we employ all-atom explicit solvent Molecular Dynamics simulations to specify the thermodynamic conditions in that the E proteins are induced to experience extensive structural changes, such as during the process of reducing pH. We study the structural behavior of the E protein monomer at acid pH solution of distinct ionic strength. Extensive simulations are carried out with all the histidine residues in its full protonated form at four distinct ionic strengths. The results are analyzed in detail from structural and energetic perspectives, and the virtual protein movements are described by means of the principal component analyses. As the main result, we found that at acid pH and physiological ionic strength, the E protein suffers a major structural change; for lower or higher ionic strengths, the crystal structure is essentially maintained along of all extensive simulations. On the other hand, at basic pH, when all histidine residues are in the unprotonated form, the protein structure is very stable for ionic strengths ranging from 0 to 225 mM. Therefore, our findings support the hypothesis that the histidines constitute the hot points that induce configurational changes of E protein in acid pH, and give extra motivation to the development of new ideas for antivirus compound design.

Immunolocalization of bone morphogenetic protein 2 during the early healing events after guided bone regeneration

Objective. The objective of this study was to evaluate the immunolocalization of bone morphogenetic protein 2 (BMP-2) after autogenous block grafting covered or not with an e-PTFE membrane. Study Design. Forty-eight rats were divided into 2 groups, autogenous block graft (B) and autogenous block graft + e-PTFE membrane (MB), and were evaluated by immunohistochemistry at baseline and 3, 7, 14, 21, and 45 days. Results. The largest number of positive cells in the recipient bed was observed after 3 days in both groups. At the graft border, the largest number of positive cells was seen after 7 days in group B and after 14 days in group MB. The highest proportion of staining in the graft was observed after 3 days in group B and after 21 days in group MB. Conclusions. High proportions of stain were related to intense revascularization and osteogenesis. Except for the interface, BMP-2 staining occurred later in group MB than in group B in all structures analyzed. (Oral Surg Oral Med Oral Pathol Oral Radiol 2012;113:533-541)