Free and total GMP (glycomacropeptide) contents of milk during bovine lactation

Amostras quinzenais, desde o parto até o final do período de lactação, obtidas de 34 vacas de três diferentes raças e propriedades, foram analisadas quanto à presença de GMP livre. Um pool das amostras quinzenais de cada rebanho foi analisada tanto para o conteúdo de GMP livre quanto para o GMP total (liberado da k-caseína pela ação da renina), correlacionando-os com as condições sanitárias do animal e do úbere, à fase da lactação e à produção de leite. A maioria dos problemas sanitários concentrou-se próximo ao parto, com poucas e espaçadas ocorrências de mastites clínicas. Os resultados do teste de CMT mostraram reações compatíveis às fases da lactação. Para o GMP livre as maiores variações ocorreram em função do período de lactação e em conseqüência de mastites clínicas e subclínicas. Valores elevados foram observados no início da lactação (5,87mg de ácido siálico/L de leite), normalizando para valores próximos de 3,30mg/L já ao final do segundo mês e voltando a elevar-se no terço final da lactação. em média, as mesmas tendências foram observadas para o teor de GMP total liberado pela ação de coalho comercial, iniciando com valores ligeiramente elevados (35,59mg/L), tornando-se normal e assim se mantendo até o sexto mês com valores próximos a 27,15mg/L, e novamente elevando-se gradualmente até o final da lactação, com 58,35mg de ácido siálico/L de leite. Esses resultados mostram-se úteis para a correta interpretação de métodos aplicados à seleção do leite, seja em relação ao status proteolítico da matéria-prima ou mesmo para coibição de fraudes por adição de soro ao leite.

Changes in sodium, potassium-ATPase induced by repeated fencamfamine: the roles of cyclic AMP-dependent protein kinase and the nitric oxide-cyclic GMP pathway

Fencamfamine (FCF) is an indirect dopamine agent with effects similar to amphetamine and cocaine. In the present study, we investigate changes in Na,K-ATPase, cyclic AMP-dependent protein kinase (PKA) and nitric oxide synthase (NOS) activity and cyclic GMP levels in the nucleus accumbens (NAc) and striatum (ST) of animals acutely or repeatedly treated with FCF (3.5 mg/kg). Na,K-ATPase had a similar activity in control and repeatedly treated animals, but was reduced in the NAc of the acute group. This enzyme was reduced in the ST in acute and repeatedly treated animals, compared to the control group. Expression of the alpha(1,2,3)-Na,K-ATPase isoforms in the NAc and the ST was not altered in all groups studied. Acute FCF induced a significant increase in PKA activity in both the ST and the NAc. Repeatedly treated animals showed a higher increase in PKA activity in the NAc, but not in the ST, when compared to the acute group. There was also an increase in both NOS activity and cyclic GMP levels only in the NAc of FCF repeatedly treated animals compared to the acute and control groups. We suggest that chronic FCF treatment is linked to a modification in Na,K-ATPase activity through the PKA and NO-cyclic GMP pathway. (C) 2003 Elsevier Ltd. All rights reserved.

Determinação de GMP e CMP* no leite por métodos espectrofotométrico (ANSM) e cromatográfico (HPLC) - parâmetros metodológicos (*glicomacropeptídeo e caseinomacropeptídeo)

Glycomacropeptide is a glycosilated fraction of bovine kappa-casein that remains soluble when milk is clotted by rennin. Determinations of milk sialic acid content are useful because its concentration reflects the amount of free GMP of milk. In normal milk these amounts are very low, 12 to 16 times lower than in sweet whey. Therefore, its determination may be applied to verify possible frauds with whey addictions, since it works as a fingerprint. With the description of a new spectrophotometric method for determination of free GMP (ANSM) occurred a simplification of procedures, being faster than others (HPLC method), without loss of accuracy. However, due to variations of glycosilation in kappa-casein between animals, during the lactation period, due to mastitis and yet due to proteolysis on milk, it was necessary to know these variations to interpret correctly the analytical results. It was analyzed 1,703 samples of producer's raw milk and 1,189 samples of processed milk (HTST and UHT). The results showed that normal milk from herd (producer's milk) have only small amounts of free GMP, with A470nm = 0.232±0.088 or 3.89±1.25 mg of sialic acid/L. The upper limit of this distribution was A = 0.496; thus every bigger value may represent a problem, being outside of normal distribution.

Search for Salmonella spp. in ostrich productive chain of Brazilian southeast region

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

Chemical properties and enzyme activity in a sewage sludge-treated soil

A soil sample was taken from the top 0-20cm at Jaboticabal county, São Paulo State, Brazil, air dried, sieved to 5mm, and placed into pots (2700g per pot). Sewage sludge was air-dried, ground to 2mm, and thoroughly mixed to the top 0-10cm soil of each pot, which were irrigated with distilled water in a total volume equivalent to the last 30years average rainfall in the region. Sorghum was sowed 120days after sewage sludge incorporation and then the irrigation was made according to the plants' requirement. When the plants were about 10 cm high, they were thinned to two per pot. Soil samples (0-10, 10-20, and 20-30 cm depth) were obtained immediately after the incorporation of sewage sludge and at 30, 60, 120, and 170 days after, air dried, sieved to 2 mm and analyzed for organic matter (OM), pH (0,01 mol L-1 CaCl2), extractable P (resin), potassium (K), calcium (Ca), and magnesium (Mg), amylase and cellulase activity. Sewage sludge increased soil OM, pH, extractable phosphorus (P), K. Ca. amylase and cellulase activity, especially at the rate 16 t ha(-1). Organic matter, extractable P, K, Ca, Mg. and amylase activity were higher in the top 0-10cm, while pH was higher in the 20-30cm layer. Amylase activity was not affected by sampling depth. Organic matter, pH, extractable P. K, Ca, and Mg decreased during the experimental period. Amylase activity decreased until sorghum was sowed and increased afterwards. Cellulase activity increased until 90 days after sewage sludge application and then decreased. Sewage sludge used in the experiment should already contain some amylase activity or a substance that was a soil enzyme activator and also a substance that was an inhibitor of soil cellulase inhibitor. Sonic of the plant nutrients contained in sewage sludge, mainly P, did not migrate down the soil column. an indication that sewage sludge should be incorporated into the soil to improve nutrient bioavailability. Sorghum roots increased amylase activity but did not affect cellulase activity.

Abordaje práctico sobre cómo mejorar el bienestar en los bovinos

Current challenges of humanity require a new paradigm for animal production, and invest time and attention to the development of new production techniques that take into account the principles of sustainability and animal welfare science. To do this, we must be committed to promoting animal welfare and health, ensure environmental sustainability, consumer satisfaction and profitability for producers. To change our relationship with animals there is no need for changes in market conditions or large investments, it is enough to know better the needs of the animals we raise, the management system adapted to its characteristics. Even in the most favorable management conditions, on farms that have technological resources and trained personnel, there is much to change, especially in the daily management of the animals. Some of the risks more evident in the traditional systems of beef production are: problems during parturition, morbidity and mortality of calves, lack of shade in the pastures, improper handling of animals, risk of injury, inadequate infrastructure, poor management practices during routine processing (marking, castration, vaccination, dehorning), poor welfare during loading, transport, unloading and slaughter. To remedy this, you can adopt good management practices, which have been shown to reduce risk and improve the welfare of cattle in different stages of the production chain. The objective of this review is to show some of the risks that reduce the welfare of cattle and examples of how the adoption of best management practices impact on improving the productivity of these production systems. Finally, some indicators for assessing welfare in production systems are shown.

Hypoxanthine-guanine phosphoribosyltransferase from Mycobacterium tuberculosis H37Rv: Cloning, expression, and biochemical characterization

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Purification and physicochemical properties of a ribonuclease produced by Aspergillus flavipes (IZ : 1501)

A RNase of Aspergillus flavipes (IZ:1501) was purified from culture medium by chromatography on DEAE-cellulose and Sephadex G50 columns, after 96 h of cultivation. The molecular weight of the RNase was estimated to be 15 kD by gel filtration using Sephadex G100, and the optimum pH and temperature were 4.0 and 55 degrees C, respectively. Catalytic activity was inhibited by Hg2+, Ag+, Fe3+, Co2+ and Zn2+. The enzyme showed guanosine specificity producing only 3'-GMP from yeast RNA.

High-fat diet associated with obesity induces impairment of mouse corpus cavernosum responses

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

Platelet hyperaggregability in high-fat fed rats: A role for intraplatelet reactive-oxygen species production

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

Three-dimensional structure of ribonuclease T-1 complexed with an isosteric phosphonate substrate analogue of GpU: Alternate substrate binding modes and catalysis

The X-ray crystal structure of a complex between ribonuclease T-1 and guanylyl(3'-6')-6'-deoxyhomouridine (GpcU) has been determined at 2.0 Angstrom resolution. This Ligand is an isosteric analogue of the minimal RNA substrate, guanylyl(3'-5')uridine (GpU), where a methylene is substituted for the uridine 5'-oxygen atom. Two protein molecules are part of the asymmetric unit and both have a GpcU bound at the active site in the same manner. The protein-protein interface reveals an extended aromatic stack involving both guanines and three enzyme phenolic groups. A third GpcU has its guanine moiety stacked on His92 at the active site on enzyme molecule A and interacts with GpcU on molecule B in a neighboring unit via hydrogen bonding between uridine ribose 2'- and 3'-OH groups. None of the uridine moieties of the three GpcU molecules in the asymmetric unit interacts directly with the protein. GpcU-active-site interactions involve extensive hydrogen bonding of the guanine moiety at the primary recognition site and of the guanosine 2'-hydroxyl group with His40 and Glu58. on the other hand, the phosphonate group is weakly bound only by a single hydrogen bond with Tyr38, unlike ligand phosphate groups of other substrate analogues and 3'-GMP, which hydrogen-bonded with three additional active-site residues. Hydrogen bonding of the guanylyl 2'-OH group and the phosphonate moiety is essentially the same as that recently observed for a novel structure of a RNase T-1-3'-GMP complex obtained immediately after in situ hydrolysis of exo-(S-p)-guanosine 2',3'-cyclophosphorothioate [Zegers et al. (1998) Nature Struct. Biol. 5, 280-283]. It is likely that GpcU at the active site represents a nonproductive binding mode for GpU [:Steyaert, J., and Engleborghs (1995) fur. J. Biochem. 233, 140-144]. The results suggest that the active site of ribonuclease T-1 is adapted for optimal tight binding of both the guanylyl 2'-OH and phosphate groups (of GpU) only in the transition state for catalytic transesterification, which is stabilized by adjacent binding of the leaving nucleoside (U) group.

Detection of DNA Nucleotides on Pretreated Boron Doped Diamond Electrodes

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

CONTROL OF ADIPOSE-TISSUE LIPOLYSIS IN ECTOTHERM VERTEBRATES

Lipolytic activity of fish (Hoplias malabaricus), toad (Bufo paracnemis), and snake (Philodryas patagoniensis) adipose tissue was investigated in vivo and in vitro. Catecholamines or glucagon did not affect the release of free fatty acids (FFA) by incubated fish and toad adipose tissue. Catecholamines also failed to activate snake adipose tissue lipolysis, which even decreased in the presence of epinephrine. However, glucagon stimulated both the lipolytic activity of reptilian tissue in vitro and the mobilization of FFA to plasma when administered to snakes in vivo. The release of FFA from incubated fish, amphibian, and reptilian adipose tissue increased markedly in the presence of cAMP or xanthine derivatives, inhibitors of phosphodiesterase. Forskolin or fluoride, activators of specific components of the adenylate cyclase system, strongly stimulated toad adipose tissue lipolysis. The data suggest that adipocyte triacylglycerol lipase of ectotherm vertebrates is activated by a cAMP-mediated phosphorylation and that the organization of the membrane-bound adenylate cyclase system is similar to that of mammals.

The Combination of Amoxicillin and Metronidazole Improves Clinical and Microbiologic Results of One-Stage, Full-Mouth, Ultrasonic Debridement in Aggressive Periodontitis Treatment

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

Human eosinophil adhesion and degranulation stimulated with eotaxin and RANTES in vitro: Lack of interaction with nitric oxide

Background: Airway eosinophilia is considered a central event in the pathogenesis of asthma. The toxic components of eosinophils are thought to be important in inducing bronchial mucosal injury and dysfunction. Previous studies have suggested an interaction between nitric oxide (NO) and chemokines in modulating eosinophil functions, but this is still conflicting. In the present study, we have carried out functional assays (adhesion and degranulation) and flow cytometry analysis of adhesion molecules (VLA-4 and Mac-1 expression) to evaluate the interactions between NO and CC-chemokines (eotaxin and RANTES) in human eosinophils. Methods: Eosinophils were purified using a percoll gradient followed byimmunomagnetic cell separator. Cell adhesion and degranulation were evaluated by measuring eosinophil peroxidase (EPO) activity, whereas expression of Mac-1 and VLA-4 was detected using flow cytometry. Results: At 4 h incubation, both eotaxin (100 ng/ml) and RANTES (1000 ng/ml) increased by 133% and 131% eosinophil adhesion, respectively. L-NAME alone (but not D-NAME) also increased the eosinophil adhesion, but the co-incubation of L-NAME with eotaxin or RANTES did not further affect the increased adhesion seen with chemokines alone. In addition, L-NAME alone (but not D-NAME) caused a significant cell degranulation, but it did not affect the CC-chemokine-induced cell degranulation. Incubation of eosinophils with eotaxin or RANTES, in absence or presence of L-NAME, did not affect the expression of VLA-4 and Mac-1 on eosinophil surface. Eotaxin and RANTES (100 ng/ml each) also failed to elevate the cyclic GMP levels above baseline in human eosinophils. Conclusion: Eotaxin and RANTES increase the eosinophil adhesion to fibronectin-coated plates and promote cell degranulation by NO-independent mechanisms. The failure of CC-chemokines to affect VLA-4 and Mac-1 expression suggests that changes in integrin function (avidity or affinity) are rather involved in the enhanced adhesion. © 2008 Lintomen et al; licensee BioMed Central Ltd.