148 resultados para MULTIMODAL ELUTION
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
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Bothrops insularis venom contains a variety of substances presumably responsible for several pharmacological effects. We investigated the biochemical and biological effects of phospholipase A(2) protein isolated from B. insularis venom and the chromatographic profile showed 7 main fractions and the main phospholipase A(2) (PLA(2)) enzymatic activity was detected in fractions IV and V. Fraction IV was submitted to a new chromatographic procedure on ion exchange chromatography, which allowed the elution of 5 main fractions designated as lV-1 to IV-5, from which lV-4 constituted the main fraction. The molecular homogeneity of this fraction was characterized by high-performance liquid chromatography (HPLC) and demonstrated by mass spectrometry (MS), which showed a molecular mass of 13984.20 Da; its N-terminal sequence presented a high amino acid identity (up to 95%) with the PLA(2) of Bothrops jararaca and Bothrops asper. Phospholipase A(2) isolated from B. insularis (Bi PLA(2)) venom (10 mu g/mL) was also studied as to its effect on the renal function of isolated perfused kidneys of Wistar rats (n = 6). Bi PLA(2) increased perfusion pressure (PP), renal vascular resistance (RVR), urinary flow (UF) and glomerular filtration rate (GFR). Sodium (%TNa+) and chloride tubular reabsorption (%TCl-) decreased at 120 min, without alteration in potassium transport. In conclusion, PLA(2) isolated from B. insularis venom promoted renal alterations in the isolated perfused rat kidney. (c) 2007 Elsevier Ltd. All rights reserved.
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O presente trabalho objetivou analisar o fluxo energético no escoamento de soja da região Centro-Oeste do Brasil, considerando as rotas atualmente existentes, mais precisamente, do município de Rio Verde -GO até o porto de Santos - SP, visto que este é um grande polo exportador de granéis sólidos. Foram selecionadas duas rotas para a análise, uma contemplando a multimodalidade, ou seja, os modais rodoviário, ferroviário e hidroviário, e outra, uma única modalidade, o modal rodoviário. A conversão dos fatores físicos e operacionais em unidades energéticas foi realizada por meio de coeficientes energéticos levantados junto à literatura. Os resultados obtidos indicaram, pela rota 1 multimodal, um gasto energético específico maior para o modal rodoviário (0,50 MJ km-1 t-1), seguido pelo modal ferroviário (0,42 MJ km-1 t-1) e, em terceiro, o modal hidroviário (0,22 MJ km-1 t-1). Pela rota 2, unimodal rodoviário, o resultado indicou 0,50 MJ km-1 t-1. Nas participações de energia apresentadas, comparando as rotas 1 e 2, a que apresentou maior gasto energético específico total foi a rota 2 (0,50 MJ km-1 t-1), seguida pela rota 1 (0,34 MJ km-1 t-1).
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Propolis obtained from honeybee hives has been widely used in medicine, cosmetics, and industry due to its versatile biological activities (antioxidant, antimicrobial, fungicidal, antiviral, antiulcer, immunostimulating, and cytostatic). These activities are mainly attributed to the presence of flavonoids in propolis, which points out the interest in quantifying these constituents in propolis preparations, as well as validation of analytical methodologies. High-performance liquid chromatography (HPLC) methods have been reported to quantify isolated flavonoids or these compounds in complex biological matrices, such as herbal raw materials and extractive preparations. An efficient, precise, and reliable method was developed for quantification of propolis extractive solution using HPLC with UV detection. The chromatograms were obtained from various gradient elution systems (GES) tested in order to establish the ideal conditions for the analysis of propolis extractive solution, using methanol and water: acetonitrile (97.5 : 2.5, v/v) as mobile phase. Gradient reversed phase chromatography was performed using a stainless steel column (250 x 4.6 mm i.d., 5 mum) filled with Chromsep RP 18 (Varian), column temperature at 30.0 +/- 0.1degreesC and detection at 310 nm. The main validation parameters of the method were also determined. The method showed linearity for chrysin in the range 0.24-2.4 mug mL(-1) with good correlation coefficients (0.9975). Precision and accuracy were determined. The obtained results demonstrate the efficiency of the proposed method. The analytical procedure is reliable and offers advantages in terms of speed and cost of reagents.
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The carotenoid composition of Brazilian Valencia orange juice was determined by open column chromatography (OCC) and high-performance liquid chromatography. Carotenoid pigments were extracted using acetone and saponified using 10% methanolic potassium hydroxide. Sixteen pigments were isolated by OCC and identified as alpha-carotene, zeta-carotene, beta-carotene, alpha-cryptoxanthin, beta-cryptoxanthin, lutein-5,6-epoxide, violaxanthin, lutein, antheraxanthin, zeaxanthin, luteoxanthin A, luteoxanthin B, mutatoxanthin A, mutatoxanthin B, auroxanthin B and trollichrome B. Thirteen carotenoid pigments were separated using a ternary gradient (acetonitrile-methanol-ethyl acetate) elution on a C-18 reversed-phase column. Among these, violaxanthin, lutein, zeaxanthin, beta-cryptoxanthin, zeta-carotene, alpha-carotene, and beta-carotene were quantified. The total carotenoid content was 12 +/- 6.7 mg/1, and the major carotenoids were lutein (23%), beta-cryptoxanthin (21%), and zeaxanthin (20%). 2005 Elsevier Ltd. All rights reserved.
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
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No presente estudo procedeu-se ao isolamento e caracterização da fração globulina majoritária (11 S) de grão-de-bico, var. IAC-Marrocos. A globulina majoritária extraída foi isolada por cromatografia de filtração em gel e de troca-iônica mostrando apenas uma banda de proteína na eletroforese em gel de poliacrilamida. A globulina majoritária, após passagem em coluna de Sephadex, revelou duas bandas protéicas de 55 e 52,5kDa e três bandas menores em gel de poliacrilamida dodecilsulfato de sódio. Na presença de 2-mercaptoetanol 6 polipeptídios na faixa de 18 a 42kDa foram revelados na eletroforese. A globulina isolada foi submetida à ação da tripsina e quimotripsina onde a forma nativa mostrou-se resistente à ação enzimática enquanto o aquecimento (96 e 121°C/15min) não foi suficiente para aumentar a susceptibilidade à hidrólise, significativamente. Adição de NaCl 0,3M levou a um aumento da estabilidade estrutural com menor susceptibilidade à digestão proteolítica, fato em parte perdido com o aquecimento. As hidrólises foram acompanhadas por eletroforese em gel de poliacrilamida dodecilsulfato de sódio.
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A germinação das sementes de grão-de-bico foi acompanhada por um período de 6 dias, no qual pequenas variações nos teores de nitrogênio e globulina total foram registradas. A globulina majoritária (tipo 11 S) apresentou maiores variações após o quarto dia de germinação. A natureza e distribuição da fração globulina majoritária isolada na cromatografia em Sepharose CL-6B mostrou pequenas modificações ao final do período de germinação. A eletroforese em gel de poliacrilamida com dodecilssulfato de sódio do pico eluído na cromatografia em Sepharose CL-6B demonstra modificações nas bandas de proteínas entre os pesos moleculares de 20 e 30 kDa e acima de 60 kDa, indicando degradação protéica durante o período. Atividade proteolítica foi detectada na fração albumina da semente que aumentou até o quarto dia, seguido de queda até o sexto dia de germinação, quando da utilização de globulina total isolada da semente e caseína como substratos. Farinha de grão-de-bico, frações albumina e globulina total isoladas não apresentaram aumento na digestibilidade in vitro; entretanto, a fração globulina majoritária isolada foi mais suscetível à hidrólise após germinação.
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
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ObjectiveTo compare the post-operative analgesic effects of butorphanol or firocoxib in dogs undergoing ovariohysterectomy.Study designProspective, randomized, blinded, clinical trial.AnimalsTwenty-five dogs > 1 year of age.MethodsDogs received acepromazine intramuscularly (IM), 0.05 mg kg-1 and either butorphanol IM, 0.2 mg kg-1 (BG, n = 12) or firocoxib orally (PO), 5 mg kg-1 (FG, n = 13), approximately 30 minutes before induction of anesthesia with propofol. Anesthesia was maintained with isoflurane. Ovariohysterectomy was performed by the same surgeon. Pain scores using the dynamic and interactive visual analog scale (DIVAS) were performed before and at 1, 2, 3, 4, 6, 8 and 20 hours after the end of surgery by one observer, blinded to the treatment. Rescue analgesia was provided with morphine (0.5 mg kg-1) IM and firocoxib, 5 mg kg-1 (BG only) PO if DIVAS > 50. Groups were compared using paired t-tests and Fisher's exact test (p < 0.05). Data are presented as mean +/- SD.ResultsThe BG required significantly less propofol (BG: 2.6 +/- 0.59 mg kg-1; FG: 5.39 +/- 0.7 mg kg-1) (p < 0.05) but the anesthesia time was longer (BG: 14 +/- 6, FG: 10 +/- 4 minutes). There were no differences for body weight (BG: 7.9 +/- 5.0, FG: 11.5 +/- 4.6 kg), sedation scores, and surgery and extubation times (BG: 10 +/- 2, 8 +/- 5 minutes; FG: 9 +/- 3, 8 +/- 4 minutes, respectively) (p > 0.05). The FG had significantly lower pain scores than the BG at 1, 2 and 3 hours following surgery (p < 0.05). Rescue analgesia was administered to 11/12 (92%) and 2/13 (15%) dogs in the BG and FG, respectively (p < 0.05).Conclusion and clinical relevanceFirocoxib produced better post-operative analgesia than butorphanol. Firocoxib may be used as part of a multimodal analgesia protocol but may not be effective as a sole analgesic.
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The analgesic efficacy of tramadol and/or vedaprofen was evaluated in cats submitted for elective ovariohysterectomy, using a randomised double blind placebo controlled design. Forty adult female cats (3.0 +/- 0.32 kg; 1.8 +/- 0.7 years) were distributed into four groups. Vedaprofen PO (0.5 mg/kg), tramadol SC (2 mg/kg), both, or placebo was administered 1 h before surgery and every 24 and 8 h, respectively, for 72 h after surgery. Pain score evaluated by interactive visual analogue and composite pain score and hyperalgesia by the von Frey filament test were recorded at 1, 2, 4, 6, 8, 12, 24, 28, 32, 48, 52, 56, 72, 96 h and on the 7th day after surgery. Animals treated with combined vedaprofen and tramadol treatment did not need rescue analgesia, did not develop hyperalgesia, and their serum cortisol concentrations and pain scores were lower than placebo until 24 and 72 h after surgery, respectively. Combined vedaprofen and tramadol treatment provided more effective postoperative analgesia and prevented hyperalgesia than when used on their own. Multimodal technique is a superior method of treating pain after feline ovariohysterectomy. This work also provides evidence for the benefits of analgesia for up to 3 days following ovariohysterectomy. (C) 2008 ESFM and AAFP. Published by Elsevier Ltd. All rights reserved.
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Objectives. To evaluate the effects of current resin-modified glass-ionomer cements (RMGICs) applied on culture of cells or implanted into subcutaneous tissue of rats.Methods. Experiment 1 - Thirty round-shaped samples of every RMGICs: Rely X Luting Cement (RL), Vitremer (VM), and Vitrebond (VB) were placed into wells with 1.1 mL of culture medium (DMEM), and incubated for 24,48 or 72 h. The extracts from every sample were applied on the MDPC-23 cells. Fresh DMEM was used as control group. The MTT assay was carried out for mitochondrial respiration. Experiment 2 - Fifty-four polyethylene tubes filled with the experimental materials were implanted into the dorsal subcutaneous tissue of rats. At 7, 30, and 90 days the animals were killed and the biopsies were processed for histological evaluation.Results. Experiment 1 - Both time of elution and material significantly influenced cell respiratory activity. in general, the extracts obtained at 24 h were less cytotoxic than 48 and 72 h incubation. The cytotoxic effect of VM and RL were not statistically different (P < 0.05) for the 24-hour period. VB showed the highest cytotoxic effect. Experiment 2 - All RMGICs elicited at 7 days a moderate to intense inflammatory reaction which decreased over time. However, connective healing occurred for most of samples at 90-day evaluation.Significance. Glass-ionomer cements may cause noticeable inflammatory response when in direct contact to connective tissue. The toxic effects of this kind of soluble material depend on the amount of components released in the aqueous environment. (C) 2005 Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved.