Potentiometric Sensor for Furosemide Determination in Pharmaceuticals, Urine, Blood Serum and Bovine Milk

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

Modified E-test by the addition of EDTA-Tris and dimethyl sulfoxide on the potentiation of the effects of some antimicrobials in Pseudomonas aeruginosa strains isolated from bovine mastitis

A concentração inibitória mínima-MIC em 30 estirpes de Pseudomonas aeruginosa isoladas de mastite bovina foi avaliada utilizando o E-test padrão e o método modificado, pela adição de Tris-EDTA e DMSO. Os métodos modificados apresentaram redução significativa da MIC das estirpes utilizando a gentamicina, a ciprofloxacina e a norfloxacina.

Effect of bovine bone morphogenetic proteins on radius fracture healing in rabbits

OBJETIVO: Investigar a influência de Proteínas Morfogenéticas Ósseas de origem bovina (bBMPs) ligadas a hidroxiapatita mais colágeno na consolidação de fraturas instáveis do rádio. MÉTODOS: em 15 coelhos com aproximadamente 5,5 meses de idade e peso médio de 3,5kg foi realizada uma fratura transversa na porção média da diáfise do rádio de ambos os membros. Na fratura do rádio direito foi aplicada mistura de bBMPs ligadas à hidroxiapatita (bBMP-HA) e colágeno bovino como aglutinante e na do rádio esquerdo, considerada controle, nenhum tratamento foi usado. Os coelhos (cinco por período) foram submetidos à eutanásia aos 30, 60 e 90 dias após a cirurgia para realização do processamento histológico e análise microscópica. RESULTADOS: A análise histológica descritiva revelou que a consolidação foi similar para os membros tratado e controle. Pela análise histomorfométrica, a área de novo osso foi em média 867442,16 mm², 938743.00 mm² e 779621,06 mm² para os membros controles e 841118,47 mm², 788038,76mm² e 618587,24 mm² para os membros tratados, aos 30, 60 e 90 dias, respectivamente. Desta forma, aos 60 dias de pós-operatório a área de novo osso foi 12.17% maior no membro tratado com bBMP-HA/colágeno em relação ao membro controle (p<0.05, teste de Tukey). em ambos os membros a área de novo osso aumentou durante o período experimental até a total consolidação da fratura. CONCLUSÃO: Baseado nos resultados obtidos foi possível concluir que a mistura de bBMP-HA/colágeno induziu pequena, porém significante melhora na consolidação da fratura.

Melatonin in maturation media fails to improve oocyte maturation, embryo development rates and DNA damage of bovine embryos

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

Endocrine profiles and ovulation rate of cows superovulated with FSH following passive immunization against steroid free-bovine follicular fluid

Dez vacas multíparas, secas, foram distribuídas aleatoriamente em dois grupos de cinco animais cada. Nos dias 8 a 12 do diestro, o primeiro grupo recebeu 100 ml de anti-soro contra líquido folicular livre de esteróides (anti-LFb) produzido em ovelhas ovariectomizadas. O segundo grupo (controle) recebeu 100 ml de soro de ovelhas não-imunizadas. Seis horas após a aplicação, os dois grupos foram superovulados com FSH (18 NIH-FSH-S1 unidades) e LH (0,29 NIH-LH-S1 unidades) administrados em quantidades decrescentes durante quatro dias. Na manhã do terceiro dia, foi administrada uma dose luteolítica de cloprostenol. Duas inseminações foram realizadas 48 e 60 horas após. Os embriões foram recuperados pelo método cervical 7 dias após a primeira inseminação. Amostras de sangue foram coletadas durante todo o período experimental para determinar, por radioimunoensaio, as concentrações plasmáticas de FSH, LH e progesterona. Todas as vacas do grupo imunizado e 3 do grupo controle apresentaram mais de 2 CL. Não existiu diferença significativa (P>0,05) na taxa de ovulação entre os grupos imunizado e controle (14,4 e 9,9, respectivamente). O número de embriões recuperado não foi significativamente diferente (P>0,05) entre os grupos, embora o grupo imunizado tenha apresentado maior número de embriões transferíveis (3,4 ± 1,0 versus 0,8 ± 0,4, P<0,05). As concentrações de gonadotrofinas plasmáticas não foram correlacionadas com a taxa de ovulação ou com o número de embriões recuperados. As concentrações de progesterona plasmática foram positivamente correlacionadas (r = 0,88, P<0,01) com a taxa de ovulação. Os resultados sugerem que o anti-LFb, aplicado antes da superovulação, não reduz a variabilidade da resposta ovariana.

QTL affecting body weight in a candidate region of cattle chromosome 5

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

Simulation and economic analysis of bovine sex selection

A simulation model implemented in the programming software Delphi XE® was applied to evaluate sex selection in bovine. The hypothesis under investigation was that a dynamic model with stochastic and deterministic elements could detect the sexed semen technique to minimize pregnancy cost and to determine the adequate number of recipients required for in vivo (ET) and in vitro embryo production (IVP) in the proposed scenarios. Sex selection was compared through semen sexed using flow cytometry (C1) and density gradient centrifugation techniques (C2) in ET and IVP. Sensibility analyses were used to identify the adequate number of recipients for each scenario. This number was reinserted into the model to determine the biological and financial values that maximized ET and IVP using sexed semen (C1M and C2M). New scenarios showed that the density gradient technique minimized pregnancy cost based on the proposed scenarios. In addition, the adequate number of recipients (ET - C1M - 115 and C2M - 105)/(IVP - C1M - 145 and C2M - 140) per donor used was determined to minimize the pregnancy cost in all scenarios.

Isolation of Candida spp. from mastitic bovine milk in Brazil

The purpose of this study was to isolate yeast (Candida) from the quarter milk of cow udders from 37 dairy farms in Brazil and to identify the different species involved in mastitis. The samples were collected between October 2002 and February 2003. Two-hundred-and-sixty milk samples from cows with clinical and subclinical mastitis were examined. Milk samples were plated onto Blood agar, Mac Conkey agar and Sabouraud dextrose agar. Forty-five (17.3%) samples were positive for the genus Candida. The Candida species isolated were C. krusei (44.5%), C. rugosa (24.5%), C. albicans (8.9%), C. guilliermondii (8.9%), and others (13.2%). We also isolated Escherichia coli (26.5%), coagulase-positive Staphylococcus (25.0%), Streptococcus spp. (8.1%), Enterobacter spp. (8.1%), and other fungi (8.1%), among others.

Role of inflammatory mediators in priming, activation, and deformability of bovine neutrophils

Objective-To determine the capacity of inflammatory mediators tumor necrosis factor-alpha (TNF-alpha), interleukin-8 (IL-8), platelet-activating factor (PAF), lipopolysaccharide (LPS), and leukotoxin to prime, activate, or alter deformability of adult bovine neutrophils.Sample Population-Blood collected from 5 healthy adult Holstein cows.Procedure-Isolated neutrophils or whole brood was incubated with TNF-alpha, IL-8, PAF, LPS, or leukotoxin, and neutrophil chemiluminescence, degranulation, deformability, shape change, CD11b expression, and size distribution was measured.Results-incubation with TNF-alpha, IL-8; PAF, and IFS primed neutrophils for oxygen radical release but caused minimal oxygen radical release by themselves. None of the inflammatory mediators induced degranulation. Incubation with TNF-alpha and PAF resulted in a decrease in neutrophil deformability and induced shape change in neutrophils. incubation with PAF consistently resulted in an increase in neutrophil size as measured by use of flow cytometry. Only IL-8 caused an increase in expression of CD11b by neutrophils.Conclusions and Clinical Relevance-Inflammatory mediators tested had minimal effects on neutrophil oxygen radical production or degranulation but did prime neutrophils for oxygen radical production. Incubation with PAF and TNF-alpha caused a decrease in neutrophil deformability and altered neutrophil shape and size. Results of our study indicate that PAF- and TNF-alpha-induced changes in neutrophil deformability and size may cause integrin- and setectin-independent trapping of neutrophils in the lungs of cattle with pneumonic pasteurellosis.

Topographical, diametral, and quantitative analysis of dentin tubules in the root canals of human and bovine teeth

The aim of this study was to evaluate the number and the diameter of dentin tubules in root canals, in the cervical, middle, and apical thirds, of human and bovine teeth. Twenty-four single-rooted, human premolars were divided into four groups (n = 6): GH1, 10 to 15 years; GH2, 16 to 30 years; GH3, 31 to 45 years; and GH4, 46 to 80 years; and 24 bovine incisors were divided into four groups (n = 6): GB1, central; GB2, lateral first; GB3, lateral second; and GB4, lateral third. The crowns were removed from the specimens, which were then debrided, sectioned longitudinally in the vestibular-lingual direction, and submitted to ultrasonic cleaning. Scanning electron microscopic evaluations were made with 1,000x and 5,000x magnification. According to the root thirds, statistically significant differences were found both for the number and the diameter of dentin tubules, with the cervical third presenting the highest mean values for both specimen types. As regards the number of dentin tubules, it was observed that the bovine specimens presented a significantly higher mean value than the human specimens; this difference was not observed when the diameters of the two types were compared.

Use of primers derived from a new sequence of the bovine Y chromosome for sexing Bos taurus and Bos indicus embryos

A bovine male-specific marker was identified in our laboratory through random amplified polymorphic DNA (RAPD) analysis. This fragment of 3216 bp was cloned, sequenced and mapped by fluorescent in situ hybridization (FISH) on the taurine Yq. Primers derived from this sequence were initially screened by polymerase chain reaction (PCR) for their ability to detect Y-specific segments in zebu and taurine genomic DNA. Two of these primers amplified a 655 bp Y-specific sequence present in taurine and zebu male genomic DNA. These primers were then used for detecting the 655 bp male sequence in DNA from 173 zebu and 30 taurine embryos, which had been previously sexed using primers for the sequence BC 1.2. The results revealed an accuracy of 100%. (C) 2002 Elsevier B.V. All rights reserved.

Fracture strength of bovine incisors after intra-radicular treatment with MTA in an experimental immature tooth model

Aim To evaluate, using an experimental immature tooth model, the fracture resistance of bovine incisors submitted to different reinforcement treatments with mineral trioxide aggregate (MTA).Methodology An immature tooth model was created by sectioning the coronal and apical portions of 40 bovine incisors 8 mm above and 12 mm below the cementoenamel junction. The root canals were irrigated with 1.0% sodium hypochlorite. They were enlarged both coronally and apically using number 703 carbide burs (ISO: 500-104-168-007-021) and their internal diameter was standardized to 2.1 mm. The specimens were assigned to four groups (n = 10): GI-control (without filling); GII-apical MTA plug + filling with gutta-percha and endodontic sealer; GIII-filling with MTA; GIV-apical MTA plug + filling with MTA + metallic post (Reforpost I). A polyether impression material was used to simulate the periodontal ligament. The specimens were submitted to a compressive load at a crosshead speed of 0.5 mm min(-1) in a servo-hydraulic universal testing machine (MTS 810) applied at 45 degrees to the long axis of the tooth until failure. Data were submitted to statistical analysis by the Kruskal-Wallis test at 5% significance level.Results GIV presented the highest fracture resistance (32.7N) and differed significantly from the other groups (P < 0.05). No statistically difference was found between GII (16.6N) and GIII (23.4N) (P > 0.05). GIII had a significantly higher fracture resistance than GI (P < 0.05).Conclusions the use of MTA + metallic post as an intra-radicular reinforcement treatment increased the resistance to fracture of weakened bovine teeth in an experimental immature tooth model.

Effect of post length on the fatigue resistance of bovine teeth restored with bonded fiber posts: A pilot study

This study evaluated the influence of the cementation length of glass fiber-reinforced composite (FRC) on the fatigue resistance of bovine teeth restored with an adhesively cemented FRC. Thirty roots of single-rooted bovine teeth were allocated to 3 groups (n = 10), according to the ratio of crown length/root length (post cementation length): group 1 = 2/3, group 2 = 1/2, and group 3 = 1/1. The roots were prepared, the fiber posts (FRC Postec Plus) were cemented, and the specimens were submitted to 2 million mechanical cycles. After fatigue testing, a score was given based on the number of fatigue cycles until fracture, and data were submitted to statistical analysis. All specimens were resistant to fatigue. Taking into account the methodology and results of this study, the evaluated fiber posts can be cemented based on the ratio of crown/root at 1/1. Further clinical studies must be conducted to verify this ratio.

Role of roscovitine and IBMX on kinetics of nuclear and cytoplasmic maturation of bovine oocytes in vitro

The 3-isobutyl-1-methylxanthine (IBMX) is able to prevent resumption of meiosis by maintaining elevated cyclic AMP (cAMP) concentrations in the oocyte, and roscovitine, a purine known to specifically inhibit MPF kinase activity, maintains bovine oocytes at the germinal vesicle (GV) stage. The present study was conducted to analyze whether cytoplasmic maturation (examined by the pattern of cortical granule (CG) distribution) of bovine oocytes is improved during meiotic arrest with IBMX and roscovitine. Oocytes were matured in vitro in a 10% Knockout(SR) supplemented TCM-199 medium (Control) with either 0.5 mM IBMX or 25 mu M roscovitine (ROSC). Oocytes were stained with fluorescein isothiocyanate conjugated Lens culinaris agglutinin (FITC-LCA) for CG evaluation and with Hoechst 33342 for nuclear stage assessment. At 16 h of culture, the percentage of oocytes remaining in the GV stage was higher (P < 0.05) in the ROSC group (32.41%) compared with the Control and IBMX groups (8.61% and 9.73%, respectively). At 24h of culture, progression of meiosis to M II stage was retarded (P < 0.05) in the ROSC group (24.05%) compared to the Control (60.20%), whereas the IBMX group (33.88%) showed no significant difference to the other two groups. At 16h of maturation, the proportion of oocytes with CG in clusters (immature cytoplasm) was similar between the groups, as was the percentage of peripheral CG (mature) at 24h of maturation. The results of the present study demonstrated that the meiotic inhibitors IBMX and roscovitine delay the progression of nuclear maturation without affecting cytoplasmic maturation, assessed by the analysis of CG repositioning. (c) 2006 Elsevier B.V. All rights reserved.