SABIIA - Sistema Aberto e Integrado de Informação em Agricultura (Sabiia)

Da-Costa-Pinto,E.A.L.; Dorsa,T.K.; Altimani,A.; Andreollo,N.A.; Cardoso,S.R.; Morais,D.J.; Bustorff-Silva,J.M..

The objective of the present study was to assess esophageal motor function in 21 children (7.5 ± 2.9 years) with caustic strictures. Esophageal manometry was performed using a water-infusion system interfaced with a polygraph and displayed on a computer screen. The data were compared with those obtained from 9 healthy children. Radionuclide transit was determined by studying deglutition of a single bolus of 99mTc pertechnetate in 10 ml of water. Non-peristaltic low-amplitude and long-duration waves were the most common findings detected in patients with strictures longer than 20% of esophageal length (N = 11). Compared with the control group, these patients presented lower mean amplitude and longer mean duration of waves (24.4 ± 11.2 vs 97.9 ± 23.7 mmHg, P...

A gasometric method to determine erythrocyte catalase activity

Palavras-chave: Caustic esophagitis; Esophageal dysfunction; Manometry; Esophageal transit.

Ano: 2004

URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2004001100005

Siqueira,A.J.S.; Remião,J.O.; Azevedo,A.M.P.; Azambuja,C.R.J..

We describe a new gasometric method to determine erythrocyte catalase activity by the measurement of the volume of oxygen produced as a result of hydrogen peroxide decomposition in a system where enzyme and substrate are separated in a special reaction test tube connected to a manometer and the reagents are mixed with a motor-driven stirrer. The position of the reagents in the test tube permits the continuous measurement of oxygen evolution from the time of mixing, without the need to stop the reaction by the addition of acid after each incubation time. The enzyme activity is reported as KHb, i.e., mg hydrogen peroxide decomposed per second per gram of hemoglobin (s-1 g Hb-1). The value obtained for catalase activity in 28 samples of hemolyzed human blood...

A glycoprotein E gene-deleted bovine herpesvirus 1 as a candidate vaccine strain

Palavras-chave: Erythrocyte; Catalase assay; Gasometry.

Ano: 1999

URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X1999000900006

Weiss,M.; Brum,M.C.S.; Anziliero,D.; Weiblen,R.; Flores,E.F..

A bovine herpesvirus 1 (BoHV-1) defective in glycoprotein E (gE) was constructed from a Brazilian genital BoHV-1 isolate, by replacing the full gE coding region with the green fluorescent protein (GFP) gene for selection. Upon co-transfection of MDBK cells with genomic viral DNA plus the GFP-bearing gE-deletion plasmid, three fluorescent recombinant clones were obtained out of approximately 5000 viral plaques. Deletion of the gE gene and the presence of the GFP marker in the genome of recombinant viruses were confirmed by PCR. Despite forming smaller plaques, the BoHV-1△gE recombinants replicated in MDBK cells with similar kinetics and to similar titers to that of the parental virus (SV56/90), demonstrating that the gE...

A high-carbohydrate diet enhances the adverse effect of the S2 allele of APOC3 SstI polymorphism on the TG/HDL-C ratio only in young Chinese females

Palavras-chave: Cattle pathogen; Immunization; Control; Differential vaccine.

Ano: 2015

URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2015000900843

Song,Yong Yan; Gong,Ren Rong; Zhang,Zhen; Li,Yuan Hao; Xiao,Li Ying; Zhou,Xue Dong; Fang,Ding Zhi.

Both genetic background and diet have profound effects on plasma lipid profiles. We hypothesized that a high-carbohydrate (high-CHO) diet may affect the ratios of serum lipids and apolipoproteins (apo) differently in subjects with different genotypes of the SstI polymorphism in the apoCIII gene (APOC3). Fifty-six healthy university students (27 males and 29 females, 22.89 ± 1.80 years) were given a washout diet of 54% carbohydrate for 7 days, followed by a high-CHO diet of 70% carbohydrate for 6 days without total energy restriction. Serum triglyceride (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), apoB100, apoAI, and the APOC3 SstI polymorphism were analyzed. The ratios of serum...

A high-carbohydrate diet induces greater inflammation than a high-fat diet in mouse skeletal muscle

Palavras-chave: High-carbohydrate diet; Lipid ratio; Gene polymorphism; Human APOC3.

Ano: 2011

URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2011000600005

Antunes,M.M.; Godoy,G.; de Almeida-Souza,C.B.; da Rocha,B.A.; da Silva-Santi,L.G.; Masi,L.N.; Carbonera,F.; Visentainer,J.V.; Curi,R.; Bazotte,R.B..

We previously reported that both the high-carbohydrate diet (HCD) and high-fat diet (HFD) given for two months promote lipid deposition and inflammation in the liver and brain of mice. The results obtained indicate a tissue-specific response to both diets. Herein, we compared the effects of HCD and HFD on fatty acid (FA) composition and inflammation in the gastrocnemius muscle. Male Swiss mice were fed with HCD or HFD for 1 or 2 months. Saturated FA (SFA), monounsaturated FA (MUFA), n-3 polyunsaturated FA (n-3 PUFA), and n-6 PUFA were quantified. The activities of stearoyl-CoA desaturase 1 (SCD-1), Δ-6 desaturase (D6D), elongase 6, and de novo lipogenesis (DNL) were estimated. As for indicators of the inflammatory tissue state, we measured myeloperoxidase...

A high-copy T7 Escherichia coli expression vector for the production of recombinant proteins with a minimal N-terminal His-tagged fusion peptide

Palavras-chave: Saturated fatty acids; Monounsaturated fatty acids; Polyunsaturated fatty acids; N-6/n-3 PUFA ratio; SFA/n-3 PUFA ratio.

Ano: 2020

URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2020000300603

Ramos,C.R.R.; Abreu,P.A.E.; Nascimento,A.L.T.O.; Ho,P.L..

We report here the construction of a vector derived from pET3-His and pRSET plasmids for the expression and purification of recombinant proteins in Escherichia coli based on T7 phage RNA polymerase. The resulting pAE plasmid combined the advantages of both vectors: small size (pRSET), expression of a short 6XHis tag at N-terminus (pET3-His) and a high copy number of plasmid (pRSET). The small size of the vector (2.8 kb) and the high copy number/cell (200-250 copies) facilitate the subcloning and sequencing procedures when compared to the pET system (pET3-His, 4.6 kb and 40-50 copies) and also result in high level expression of recombinant proteins (20 mg purified protein/liter of culture). In addition, the vector pAE enables the expression of a fusion...

A high-fructose diet induces changes in pp185 phosphorylation in muscle and liver of rats

Palavras-chave: Escherichia coli; Expression vector; Immobilized metal affinity chromatography; Protein purification.

Ano: 2004

URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2004000800001

Ueno,M.; Bezerra,R.M.N.; Silva,M.S.; Tavares,D.Q.; Carvalho,C.R.; Saad,M.J.A..

Insulin stimulates the tyrosine kinase activity of its receptor resulting in the tyrosine phosphorylation of pp185, which contains insulin receptor substrates IRS-1 and IRS-2. These early steps in insulin action are essential for the metabolic effects of insulin. Feeding animals a high-fructose diet results in insulin resistance. However, the exact molecular mechanism underlying this effect is unknown. In the present study, we determined the levels and phosphorylation status of the insulin receptor and pp185 (IRS-1/2) in liver and muscle of rats submitted to a high-fructose diet evaluated by immunoblotting with specific antibodies. Feeding fructose (28 days) induced a discrete insulin resistance, as demonstrated by the insulin tolerance test. Plasma...

A high-fructose diet induces insulin resistance but not blood pressure changes in normotensive rats

Palavras-chave: Fructose; Insulin receptor; Pp185 phosphorylation.

Ano: 2000

URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2000001200004

Bezerra,R.M.N.; Ueno,M.; Silva,M.S.; Tavares,D.Q.; Carvalho,C.R.O.; Saad,M.J.A.; Gontijo,J.A.R..

Rats fed a high-fructose diet represent an animal model for insulin resistance and hypertension. We recently showed that a high-fructose diet containing vegetable oil but a normal sodium/potassium ratio induced mild insulin resistance with decreased insulin receptor substrate-1 tyrosine phosphorylation in the liver and muscle of normal rats. In the present study, we examined the mean blood pressure, serum lipid levels and insulin sensitivity by estimating in vivo insulin activity using the 15-min intravenous insulin tolerance test (ITT, 0.5 ml of 6 µg insulin, iv) followed by calculation of the rate constant for plasma glucose disappearance (Kitt) in male Wistar-Hannover rats (110-130 g) randomly divided into four diet groups: control, 1:3 sodium/potassium...

A key role for Na+/K+-ATPase in the endothelium-dependent oscillatory activity of mouse small mesenteric arteries

Palavras-chave: High-fructose diet; Insulin resistance; Blood pressure; Hypertriglyceridemia; Plasma cholesterol.

Ano: 2001

URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2001000900008

Giachini,F.R.C.; Carneiro,F.S.; Lima,V.V.; Carneiro,Z.N.; Brands,M.W.; Webb,R.C.; Tostes,R.C..

Oscillatory contractile activity is an inherent property of blood vessels. Various cellular mechanisms have been proposed to contribute to oscillatory activity. Mouse small mesenteric arteries display a unique low frequency contractile oscillatory activity (1 cycle every 10-12 min) upon phenylephrine stimulation. Our objective was to identify mechanisms involved in this peculiar oscillatory activity. First-order mesenteric arteries were mounted in tissue baths for isometric force measurement. The oscillatory activity was observed only in vessels with endothelium, but it was not blocked by L-NAME (100 µM) or indomethacin (10 µM), ruling out the participation of nitric oxide and prostacyclin, respectively, in this phenomenon. Oscillatory activity was not...

A laboratory cage for foster nursing newborn mice

Palavras-chave: Oscillatory activity; Na+/K+-ATPase pump; Ouabain; Small mesenteric arteries.

Ano: 2009

URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2009001100010

Marques-de-Araújo,S.; Cardoso,M.A..

We describe a cage to be used for foster nursing in order to guarantee that original mother's colostrum is not ingested by the newborn mice. A common (30.5 cm x 19.5 cm x 12.0 cm) mouse cage was fitted with a wire net tray with a mesh (1 cm x 1 cm), which divides the cage into an upper and a lower compartment. Mice born to females placed in the upper compartment pass through the mesh and fall into the lower compartment, where another lactating female with one or two of its own pups are. Of a total of 28 newborn mice of C3H/He and Swiss strains, 23 were successfully fostered. Important observations are presented to show that this is a valuable alternative for foster studies without great suffering on the part of the female.

A leucine-supplemented diet improved protein content of skeletal muscle in young tumor-bearing rats

Palavras-chave: Foster nursing techniques; Foster mother; Cage for fostering; Fostering.

Ano: 1999

URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X1999000300011

Gomes-Marcondes,M.C.C.; Ventrucci,G.; Toledo,M.T.; Cury,L.; Cooper,J.C..

Cancer cachexia induces host protein wastage but the mechanisms are poorly understood. Branched-chain amino acids play a regulatory role in the modulation of both protein synthesis and degradation in host tissues. Leucine, an important amino acid in skeletal muscle, is higher oxidized in tumor-bearing animals. A leucine-supplemented diet was used to analyze the effects of Walker 256 tumor growth on body composition in young weanling Wistar rats divided into two main dietary groups: normal diet (N, 18% protein) and leucine-rich diet (L, 15% protein plus 3% leucine), which were further subdivided into control (N or L) or tumor-bearing (W or LW) subgroups. After 12 days, the animals were sacrificed and their carcass analyzed. The tumor-bearing groups showed a...

A link between sleep loss, glucose metabolism and adipokines

Palavras-chave: Leucine; Amino acid; Walker 256 carcinosarcoma; Body composition; Skeletal muscle protein.

Ano: 2003

URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2003001100017

Padilha,H.G.; Crispim,C.A.; Zimberg,I.Z.; De-Souza,D.A.; Waterhouse,J.; Tufik,S.; de-Mello,M.T.

The present review evaluates the role of sleep and its alteration in triggering problems of glucose metabolism and the possible involvement of adipokines in this process. A reduction in the amount of time spent sleeping has become an endemic condition in modern society, and a search of the current literature has found important associations between sleep loss and alterations of nutritional and metabolic contexts. Studies suggest that sleep loss is associated with problems in glucose metabolism and a higher risk for the development of insulin resistance and type 2 diabetes mellitus. The mechanism involved may be associated with the decreased efficacy of regulation of the hypothalamus-pituitary-adrenal axis by negative feedback mechanisms in...

A liquid phase blocking ELISA for the detection of antibodies against infectious bronchitis virus

Palavras-chave: Glucose metabolism; Insulin resistance; Sleep loss; Adipokines; Type 2 diabetes mellitus; Inflammatory status.

Ano: 2011

URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2011001000003

Cardoso,T.C.; Mouro-Sousa,R.L.; Oliveira,C.; Stringhini,G.; Augusto-Pinto,A..

A liquid phase blocking ELISA (LPB-ELISA) was developed for the detection and measurement of antibodies against infectious bronchitis virus (IBV). The purified and nonpurified virus used as antigen, the capture and detector antibodies, and the chicken hyperimmune sera were prepared and standardized for this purpose. A total of 156 sera from vaccinated and 100 from specific pathogen-free chickens with no recorded contact with the virus were tested. The respective serum titers obtained in the serum neutralization test (SNT) were compared with those obtained in the LPB-ELISA. There was a high correlation (r2 = 0.8926) between the two tests. The LPB-ELISA represents a single test suitable for the rapid detection of antibodies against bronchitis virus in...

A liver metalloendopeptidase which degrades the circulating hypotensive peptide hormones bradykinin and atrial natriuretic peptide

Palavras-chave: Liquid phase blocking ELISA; Humoral response; Infectious bronchitis virus.

Ano: 1999

URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X1999000600010

Carvalho,K.M.; Nava,R.A.; França,M.S.F.; Medeiros,M.A.S.; Camarão,G.C.; Juliano,L..

A new metalloendopeptidase was purified to apparent homogeneity from a homogenate of normal human liver using successive steps of chromatography on DEAE-cellulose, hydroxyapatite and Sephacryl S-200. The purified enzyme hydrolyzed the Pro7-Phe8 bond of bradykinin and the Ser25-Tyr26 bond of atrial natriuretic peptide. No cleavage was produced in other peptide hormones such as vasopressin, oxytocin or Met- and Leu-enkephalin. This enzyme activity was inhibited by 1 mM divalent cation chelators such as EDTA, EGTA and o-phenanthroline and was insensitive to 1 µM phosphoramidon and captopril, specific inhibitors of neutral endopeptidase (EC 3.4.24.11) and angiotensin-converting enzyme (EC 3.4.15.1), respectively. With Mr 85 kDa, the enzyme exhibits optimal...

A low concentration of ouabain (0.18 µg/kg) enhances hypertension in spontaneously hypertensive rats by inhibiting the Na+ pump and activating the renin-angiotensin system

Palavras-chave: Liver metalloendopeptidase; Bradykinin; Atrial natriuretic peptide.

Ano: 1999

URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X1999000100007

Siman,F.D.M.; Stefanon,I.; Vassallo,D.V.; Padilha,A.S..

We investigated the effects of low ouabain concentrations on systolic (SAP) and diastolic (DAP) arterial pressures and on pressor reactivity in 3-month-old male spontaneously hypertensive rats (SHR). Arterial blood pressure (BP) and pressor reactivity to phenylephrine (PHE) were investigated before and after 0.18 μg/kg ouabain administration (N = 6). The influence of hexamethonium (N = 6), canrenone (N = 6), enalapril (N = 6), and losartan (N = 6) on ouabain actions was evaluated. Ouabain increased BP (SAP: 137 ± 5.1 to 150 ± 4.7; DAP: 93.7 ± 7.7 to 116 ± 3.5 mmHg; P<0.05) but did not change PHE pressor reactivity. Hexamethonium reduced basal BP in control but not in ouabain-treated rats. However, hexamethonium + ouabain increased DAP sensitivity to...

A majority of Brazilian patients with rheumatoid arthritis HLA-DRB1 alleles carry both the HLA-DRB1 shared epitope and anti-citrunillated peptide antibodies

Palavras-chave: Ouabain; Hypertension; Na+ pump; Renin-angiotensin system.

Ano: 2010

URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2010000800010

Louzada-Júnior,P.; Freitas,M.V.C.; Oliveira,R.D.R.; Deghaide,N.H.S.; Conde,R.A.; Bertolo,M.B.; Donadi,E.A..

The objective of the present study was to evaluate the contribution of the shared epitope (SE), the rheumatoid arthritis (RA) protection model, and the occurrence of anti-cyclic citrullinated peptide (anti-CCP) antibodies in RA patients from a genetically diverse population. One hundred and forty Brazilian RA patients and 161 matched controls were typed for HLA-DRB1 alleles using amplified DNA hybridized with sequence-specific oligonucleotide probes or primers. Patients were stratified according to the presence or absence of SE (DRB1*0401, *0404, *0405, *0101, *1001, and *1402), of the DERAA alleles (DRB1*0103, *0402, *1102, *1103, *1301, *1302, and *1304), and X (all other alleles). Anti-CCP antibodies were measured by ELISA. The combined frequency of...

A mathematical framework for group analysis of von Willebrand factor multimeric composition following luminography

Palavras-chave: Susceptibility to rheumatoid arthritis; HLA; Brazilians; Shared epitope; Anti-cyclic citrullinated peptide.

Ano: 2008

URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2008000600010

Lopes,A.A.; Soares,R.P.S.; Maeda,N.Y..

The objective of the present study was to establish a method for quantitative analysis of von Willebrand factor (vWF) multimeric composition using a mathematical framework based on curve fitting. Plasma vWF multimers from 15 healthy subjects and 13 patients with advanced pulmonary vascular disease were analyzed by Western immunoblotting followed by luminography. Quantitative analysis of luminographs was carried out by calculating the relative densities of low, intermediate and high molecular weight fractions using laser densitometry. For each densitometric peak (representing a given fraction of vWF multimers) a mean area value was obtained using data from all group subjects (patients and normal individuals) and plotted against the distance between the peak...

A mechanistic view of mitochondrial death decision pores

Palavras-chave: Von Willebrand factor; Luminography; Densitometry; Curve fitting; Pulmonary hypertension.

Ano: 2002

URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2002001100002

Belizário,J.E.; Alves,J.; Occhiucci,J.M.; Garay-Malpartida,M.; Sesso,A..

Mitochondria increase their outer and inner membrane permeability to solutes, protons and metabolites in response to a variety of extrinsic and intrinsic signaling events. The maintenance of cellular and intraorganelle ionic homeostasis, particularly for Ca2+, can determine cell survival or death. Mitochondrial death decision is centered on two processes: inner membrane permeabilization, such as that promoted by the mitochondrial permeability transition pore, formed across inner membranes when Ca2+ reaches a critical threshold, and mitochondrial outer membrane permeabilization, in which the pro-apoptotic proteins BID, BAX, and BAK play active roles. Membrane permeabilization leads to the release of apoptogenic proteins: cytochrome c, apoptosis-inducing...

A meta-analysis of probiotics for preventing necrotizing enterocolitis in preterm neonates

Palavras-chave: Mitochondrial outer membrane permeabilization; Permeability transition pore; Cytochrome c; Reactive oxygen species; Caspases; BCL-2.

Ano: 2007

URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2007000800001

Yang,Y.; Guo,Y.; Kan,Q.; Zhou,X.G.; Zhou,X.Y.; Li,Y..

Necrotizing enterocolitis (NEC) is one of the most common acquired diseases of the gastrointestinal tract in preterm infants. Some randomized, controlled trials (RCTs) have indicated that probiotics may potentially lower the incidence of NEC and mortality. However, debate still remains about the safety of probiotics and their influence on normal infant growth. We performed this meta-analysis to assess the safety and benefits of probiotic supplementation in preterm infants. We searched in PubMed, Embase, and Cochrane databases for English references, and in Wanfang, VIP, and CNKI databases for Chinese references. Ultimately, 27 RCTs (including 9 Chinese articles) were incorporated into this meta-analysis. Relative risk (RR) and weighted mean difference...

A method for multiple sequential analyses of macrophage functions using a small single cell sample

Palavras-chave: Necrotizing enterocolitis; Probiotics; Meta-analysis; Preterm.

Ano: 2014

URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2014000900804

Nascimento,F.R.F.; Rodríguez,D.; Gomes,E.; Fernvik,E.C.; Russo,M..

Microbial pathogens such as bacillus Calmette-Guérin (BCG) induce the activation of macrophages. Activated macrophages can be characterized by the increased production of reactive oxygen and nitrogen metabolites, generated via NADPH oxidase and inducible nitric oxide synthase, respectively, and by the increased expression of major histocompatibility complex class II molecules (MHC II). Multiple microassays have been developed to measure these parameters. Usually each assay requires 2-5 x 10(5) cells per well. In some experimental conditions the number of cells is the limiting factor for the phenotypic characterization of macrophages. Here we describe a method whereby this limitation can be circumvented. Using a single 96-well microassay and a very small...