Peroxidase from peach fruit: Thermal stability

Peroxidase from peach fruit was purified 28.9-fold by DEAE-cellulose, Sephadex G-100 and hydroxylapatite chromatography. The purified enzyme showed only one peak of activity with an optimum pH of 5.0 and temperature of 40 degreesC. The calculated activation energy (Ea) for the reaction was 7.97 kcal/mol. The enzyme was heat-labile in the temperature range of 60 to 80 degreesC with a fast inactivation at 80 degreesC. PAGE of the inactivation course at 70 degreesC showed only one band of activity. Different sugars increased the heat stability of the activity in the following order: sucrose>lactose>glucose>fructose. Measurement of residual activity showed a stabilizing effect of sucrose at various temperature/sugar concentrations (10 to 40%, w/w) with the Ea for inactivation increasing with sucrose concentration from 0 to 20% (w/w). After inactivation at 70 degreesC and 75 degreesC the enzyme was able to be reactivated by up to 40% of the initial activity when stared at 30 degreesC.

Potentiometric determination of saccharin in commercial artificial sweeteners using a silver electrode

A simple, precise, rapid and low-cost potentiometric method for saccharin determination in commercial artificial sweeteners is proposed. Saccharin present in several samples of artificial sweeteners is potentiometrically titrated with silver nitrate solution using a silver wire as the indicator electrode, coupled to a titroprocessor. The best pH range was from 3.0 to 3.5 and the detection limit of sodium saccharin was 2.5 mg/ml. Substances normally found along with saccharin in several commercial artificial sweeteners such as maltodextrin, glucose, sucrose, fructose, aspartame, cyclamate, caffeine, sorbitol, lactose, nitrate, methyl- and n-propyl-p-hydroxybenzoate, benzoic, citric and ascorbic acids do not interfere even in significant amounts (e.g. 20 excess relative to saccharin). Chloride ion interferes when present in concentrations larger than 10 mg l(-1); this interference is eliminated with previous extraction of the sweetener from the aqueous medium with ethyl acetate. The results obtained by applying the proposed method compared very favorably with those given by the HPLC method recommended by the FDA. (C) 2003 Elsevier Ltd. All rights reserved.

Potentiometric determination of alendronate in pharmaceutical formulations

A simple, precise, fast and low-cost potentiometric method for alendronate determination in pharmaceutical formulations has been proposed. Alendronate present in tablets at the known concentration was potentiometrically titrated in an aqueous solution using a combined glass electrode coupled with an autotitrator. Determination limit of alendronate was 20.0 mug mL(-1). In the presence of common excipients, such as lactose, microcrystalline cellulose, glucose, croscarmellose sodium, starch, talc, magnesium stearate and titanium dioxide no interferences were observed. The method proposed was validated by determining alendronate in various dosage forms. The average RSD was 0.75%.

Efeito de Diferentes Diluídores na Fertilidade de Éguas de Diferentes Haras Inseminadas com Sêmen Diluído, Resfriado e Transportado em Conteiner Especial

The effects of extender and farms on the fertility of sixty-five Mangalarga Marchador breed mares were studied. The mares were randomly assigned to a 2 x 2 factorial experiment, with two breeding farms (H1 and H2) and two extenders (glycine-egg yolk-T1 and lactose-egg yolk modified-T2), from only one stallion, with different number of replicates. The mares were inseminated, on alternated days, with diluted, cooled, transported semen, in a special container (MSP-2). There were no differences in the conception rate at first cycle, regardless of the breeding farm or extender used. Also. there was no difference among breeding farm or extender concerning the conception rate/cycle and the following reproductive characteristics: number of cycles/pregnancy, number of inseminations/mare, inseminations/pregnant mare, inseminations/open mare, water temperature in the container, temperature of semen in the container, time between collection and insemination and efficiency of pregnancy. The use of lactose-egg yolk modified extender for the dilution, cooling and transport of stallion semen between farms is recommended. The new container MSP-2 showed to be a option for the transportation between farms, for the same purpose.

Effect of beta-lactoglobulin polymorphism and seasonality on bovine milk composition

The objective was to evaluate the effect of beta-lactoglobulin (beta-lg) polymorphism and seasonality on milk composition (fat, lactose, total solids, milk urea nitrogen, total protein, true protein, casein and somatic cell counts) of Holstein and Girolando cows. Milk and blood samples from 278 Holsteins cows and 156 Girolando cows were taken during two dry seasons and two rainy seasons, for milk composition analysis and to determine beta-lg genotypes, respectively. BB genotype was the most frequent for both breeds, followed by AA genotype for Holstein (BB>AA>AB) and by AB for Girolando cows (BB>AB>AA). No differences were found in milk compositional characteristics among genetic variants of beta-lg (AA, AB and BB) either between Holstein or Girolando cows. No association between milk composition and beta-lg genetic polymorphism was observed. During the dry season, independently of the breed considered, higher contents of lactose, true protein, casein and casein :true protein ratio were found.

Location of the β-galactosidase of the yeast Kluyveromyces marxianus var. marxianus ATCC 10022

During the growth of Kluyveromyces marxianus var. marxianus ATCC 10022 on lactose, peaks of glucose, but not β-galactosidase activity, were detected in culture medium. Harvested and washed whole cells produced glucose and galactose from lactose, or ortho-nitro-phenol from the chromogenic substrate ortho-nitro-phenyl-β-D-galactopyranoside (ONPG), indicating that β-galactosidase is physically associated with cells. ONPG hydrolysis by whole cells presented a monophasic kinetics (Km 36.6 mM) in lactose exponential growth phase cells, but a biphasic kinetics (Km 0.2 and 36.6 mM) in stationary growth phase cells. Permeabilization with digitonin or disruption of cells from both growth phases led to monosite ONPG hydrolysis (Km 2.2 to 2.5 mM), indicating that β-galactosidase is not located in the periplasm. In addition, the energy inhibitors fluoride or arsenate, as well as the uncouplercarbonyl cyanide m-chlorophenylhydrazone (CCCP) prevented ONPG hydrolysis by whole cells. These findings indicate that energy coupled transmembrane transport is the rate-limiting step for intracellular ONPG cleavage. The taxonomic and physiologic implications of the exclusive intracellular location of β-galactosidase of K. marxianus var. marxianus ATCC 10022 are discussed. © 1996 Kluwer Academic Publishers.

Antibiotic 26-deoxylaidlomycin isolated from Streptomyces sp. Ar386 from Brazilian soil

An actinomycete strain (Ar386) was isolated from the soil of the Araraquara regio, SP, Brazil. The strain, named Streptomyces jacareensis, formed irregular rayed, rugose, grayish-white mycelium with sinuous, branched hyphae carrying rare isolated spores; assimilated glucose, galactose, inositol, ribose, maltose, sucrose, melibiose and starch but not mannitol, rhamnose, arabinose, xylose, lactose and raffinose; and contained LL- diaminopimelic acid in its cell wall. An antibiotic active against Gram- positive bacteria, which was characterized as being 26-deoxylaidlomycin and which may have application against poultry coccidiosis, was isolated from cultures of the strain. This was the first isolation of this antibiotic from a microorganism of the genus Streptomyces and also the first isolation of this antibiotic in Brazil.

β-galactosidase immobilization on controlled pore silica

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

Control of amylase production and growth characteristics of Aspergillus ochraceus

The growth and the extracellular amylase production by Aspergillus ochraceus were studied in a stationary culture medium. Maximum growth rate of this fungus was found after 5 days of incubation at 30° C, but maximum amylase production was obtained after 2 days. The highest amylase production were attained with lactose, maltose, xylose and starch as carbon sources. The extracellular amylase production and mycelial growth were influenced by the concentration of starch. Other carbohydrates supported growth but did not induce amylase synthesis and glucose repressed it, indicating catabolite repression in this microorganism. The presence of both mechanisms of induction and repression suggests that at least these multiple forms of regulation are present in A. ochraceus. Of the nitrogen sources tested, casaminoacids, ammonium nitrate and sodium nitrate stimulated the highest yield of amylase. Optimal amylase production was obtained at pH 5.0, but enzyme activity was found only in the 4.0-6.0 pH range. These results were probably due to the inhibitory effect of NH 4 +-N in the culture medium.

Factors affecting somatic cell counts and their relations with milk and milk constituent yield in buffaloes

Data concerning daily milk yield (MY), percentage of milk fat (%F), protein (%P), lactose (%LT), and total solids (%TS), and somatic cell counts (SCC) for a herd of 222 Murrah buffalo reared in the state of São Paulo, Brazil, were collected monthly from 1997 to 2000 in order to study the factors affecting SCC and their relation to milk production and constituents during lactation. SCC decreased in the second month of lactation and increased thereafter, up to the ninth month of lactation. The interaction of month of lactation x order of calving was significant. Mean MY observed during the first month of lactation was 6.87 kg, which increased to 7.65 kg during the second month, and then decreased until the ninth month of lactation (3.83 kg). During the different months of lactation, %F, %P, %LT, and %TS ranged from 6.28 to 8.38%, 4.05 to 4.59%, 4.96 to 5.34%, and 16.94 to 18.55%, respectively. Calving year, calving order, and order of month of lactation significantly affected MY, %F, %P, %LT, and %TS. The regression coefficients of transformed SCC on MY and %LT were negative and significant during all months of lactation, showing that milk and lactose yield decreased with increased transformed SCC, causing losses to buffalo milk producers.

Evaluation of adverse effects of long-term oral administration of carprofen, etodolac, flunixin meglumine, ketoprofen, and meloxicam in dogs

Objective - To evaluate adverse effects of long-term oral administration of carprofen, etodolac, flunixin meglumine, ketoprofen, and meloxicam in dogs. Animals - 36 adult dogs. Procedures - Values for CBC, urinalysis, serum biochemical urinalyses, and occult blood in feces were investigated before and 7, 30, 60, and 90 days after daily oral administration (n = 6 dogs/group) of lactose (1 mg/kg, control treatment), etodolac (15 mg/kg), meloxicam (0.1 mg/kg), carprofen (4 mg/kg), and ketoprofen (2 mg/kg for 4 days, followed by 1 mg/kg daily thereafter) or flunixin (1 mg/kg for 3 days, with 4-day intervals). Gastroscopy was performed before and after the end of treatment. Results - For serum γ-glutamyltransferase activity, values were significantly increased at day 30 in dogs treated with lactose, etodolac, and meloxicam within groups. Bleeding time was significantly increased in dogs treated with carprofen at 30 and 90 days, compared with baseline. At 7 days, bleeding time was significantly longer in dogs treated with meloxicam, ketoprofen, and flunixin, compared with control dogs. Clotting time increased significantly in all groups except those treated with etodolac. At day 90, clotting time was significantly shorter in flunixin-treated dogs, compared with lactose-treated dogs. Gastric lesions were detected in all dogs treated with etodolac, ketoprofen, and flunixin, and 1 of 6 treated with carprofen. Conclusions and Clinical Relevance - Carprofen induced the lowest frequency of gastrointestinal adverse effects, followed by meloxicam. Monitoring for adverse effects should be considered when nonsteroidal anti-inflammatory drugs are used to treat dogs with chronic pain.

Fatty acid production by four strains of Mucor hiemalis grown in plant oil and soluble carbohydrates

Four Mucor hiemalis strains (M1, M2, M3 and M4), isolated from soil at a depth of 0 - 15 cm in the Juréia-Itatins Ecology Station (JIES), in the state of São Paulo, Brazil and were evaluated for the production of γ-linolenic (GLA) and other unsaturated fatty acids. Five growth variables (temperature, pH, carbon source, nitrogen source, and vegetable oils) were studied. Liquid media containing 2% vegetable oil (palm oil, canola oil, soybean oil, sesame oil, or sunflower oil) or 2% carbohydrate (fructose, galactose, glycerol, glucose, lactose, maltose, sucrose, sorbitol or xylose) and 1% yeast extract as a nitrogen source were used. The greatest biomass production was observed with M3 and M4 strains in palm oil (91.5 g l -1) and sunflower oil (68.3 g l -1) media, respectively. Strain M4 produced greater quantities of polyunsaturated acids in medium containing glucose. The GLA production in the M4 biomass was 1,132.2 mg l -1 in glucose medium. Plant oils were inhibitors of fatty acid production by these strains. © 2007 Academic Journals.

Comparison of β-1,3-glucanase production by Botryosphaeria rhodina MAMB-05 and Trichoderma harzianum Rifai and its optimization using a statistical mixture-design

Botryosphaeria rhodina MAMB-05 produced β-1,3-glucanases and botryosphaeran when grown on glucose, while Trichoderma harzianum Rifai only produced the enzyme. A comparison of long-term cultivation (300h) by B. rhodina demonstrated a correlation between the formation of botryosphaeran (48h) and its consumption (after 108h), and de-repression of β-1,3-glucanase synthesis when glucose was depleted from the nutrient medium, whereas for T. harzianum enzyme production commenced during exponential growth. Growth profiles and levels of β-1,3-glucanases produced by both fungi on botryosphaeran also differed, as well as the production of β-1,3-glucanases and β-1,6-glucanases on glucose, lactose, laminarin, botryosphaeran, lasiodiplodan, curdlan, Brewer's yeast powder and lyophilized fungal mycelium, which were dependent upon the carbon source used. A statistical mixture-design used to optimize β-1,3-glucanase production by both fungi evaluated botryosphaeran, glucose and lactose concentrations as variables. For B. rhodina, glucose and lactose promoted enzyme production at the same levels (2.30UmL -1), whereas botryosphaeran added to these substrates exerted a synergic effect favorable for β-glucanase production by T. harzianum (4.25UmL -1). © 2010 Elsevier B.V.

Maltodextrina em rações de leitões desmamados com diferentes pesos: Desempenho e morfometria intestinal

Two trials were conducted: in the first trial, 96 weaning pigs at the age of 21 days were used, in a block design, with a 2 x 2 factorial arrangement (two sources of carbohydrate and two weaning weight classes). In the second trial, 48 pigs were used, in a block design, with 2 x 2 x 2 x 3 factorial arrangement (two weaning weight classes x two sources of carbohydrate x two portions of the small intestine x three slaughter periods). The effects of the replacement of lactose by maltodextrin in diets of weaning pigs, with different weights, were evaluated on daily feed intake, daily weight gain, feed:gain ratio, villus height, crypt depth and thickness of the mucosa. In the first trial, from 0 to 17 days and from 0 to 29 days after weaning, effects of weight classes were detected on daily weight gain and daily feed intake. In the second trial, maltodextrin resulted in an increase in the average thickness of the intestinal mucosa and in average crypt depth, only in lighter animals. Maltodextrin can be used as an alternative to lactose in diets of weaning pigs.

Estudos de pré-formulação de cápsulas gelatinosas duras contendo formulação de extrato de Gymnema sylvestre em pó

Gymnema sylvestre extract (GSE) is a plant product widely used as an adjuvant in the treatment of diabetes mellitus and commercially available as a powder. Owing to its low flowability, the manufacturing of hard gelatin capsules containing GSE faces specific problems. The purpose of this study was to investigate the best excipient (starch, lactose or microcrystalline cellulose) for hard gelatin capsules containing GSE. The technological properties such us bulk density (ρβ); tapped density (ρt); inter-particle porosity (Ie); Carr index (CI); Hausner ratio (HR); loss on drying (%LOD) and particle size distribution (%Pf) of the various GSE mixtures were investigated with the aim of identifying the best excipient. The need for lubricants (talc/magnesium stearate) was also assessed. GSE was characterized as a fine powder with more than 50% of its particles between 0.149mm to 0.250mm; furthermore, CI=25.6%; RH=1.3 and Ie = 25.6% and, as expected with such properties, it showed poor flowability. All the excipients investigated were able to change the technological properties of GSE and the powder mixture containing microcrystalline cellulose gave the best results.