Beta-glucosidase from Chalara paradoxa CH32: purification and properties

The hyphomycete Chalara paradoxa CH32 produced an extracellular beta-glucosidase during the trophophase. The enzyme was purified to homogeneity by ion-exchange and size-exclusion chromatography. The purified enzyme had an estimated molecular mass of 170 kDa by size-exclusion chromatography and 167 kDa by SDS-PAGE. The enzyme had maximum activity at pH 4.0-5.0 and 45 degrees C. The enzyme was inactivated at 60 degrees C. At room temperature, it was unstable at acidic pH, but it was stable to alkaline pH. The purified enzyme was inhibited markedly by Hg(2+) and Ag(2+) and also to some extent by the detergents SDS, Tween 80, and Triton X-100 at 0.1%. Enzyme activity increased by 3-fold in the presence of 20% ethanol and to a lesser extent by other organic solvents. Purified beta-glucosidase was active against cellobiose and p-nitrophenyl-beta-D-glucopyranoside but did not hydrolyze lactose, maltose, sucrose, cellulosic substrates, or galactopyranoside, mannopyranoside, or xyloside derivatives of p-nitrophenol. The V(max) of the enzyme for p-NPG (K(m) = 0.52 mM) and cellobiose (K(m) = 0.58 mM) were 294 and 288.7 units/mg, respectively. Hydrolysis of pNPG was inhibited competitively by glucose (K(i) = 11.02 mM). Release of reducing sugars from carboxymethylcellulose by a purified endoglucanase produced by the same organism increased markedly in the presence of beta-glucosidase.     

Intervertebral Disc Protrusion in a Ferret with Triple Thoracic Block Vertebrae

Hind limb paresis or plegia in ferrets is a common sign of illness, which can be produced by intervertebral disc (IVD) extrusion or protrusion along with many other systemic diseases. This case report describes a ferret (Mustela putorius furo) with chronic onset paraplegia in which T7-T8, T8-T9, and T10-T11 IVD herniations and T9-T10 block vertebrae were diagnosed with the assistance of myelography. It is believed that the excessive stress on the adjacent annulus fibrosus and vertebral endplates resulting from block vertebrae could have been the cause of IVD disease in the atypical location described in this case.     

Rupture of gastrocnemius tendon in broiler breeder hens

Lameness and mortality of 0.55% to 1.00% per week occurred in 32-to-33-wk-old broiler breeder hens in a complex involving eight houses. Males were not affected. Laboratory evaluation of 19 birds, over 2 wk, revealed complete rupture or partial tear, primarily of the gastrocnemius tendon; the fibularis longus, fibularis brevis, and flexor muscles and their tendons were injured in a few birds. Rupture of the gastrocnemius tendon has been observed in broiler breeder chickens older than 12 wk of age and it is characterized by lameness secondary to unilateral or bilateral rupture of the tendon. The cause of this condition has been primarily attributed to reovirus infection but rupture by other causes can also occur. In this case, there was no evidence of synovitis or reovirus infection in the birds. The cause of the ruptures was attributed to aggressive males when the ratio of males to females was increased from seven or eight males per 100 females at 22-23 wk to nine or 10 per 100 at 31 wk of age.     

Cell wall polysaccharides of near-isogenic lines of melon (Cucumis melo L.) and their inbred parentals which show differential flesh firmness or physiological behavior

We characterized differences in cell wall material and polysaccharide structures, due to the quantitative trait loci associated with higher flesh firmness in a nonclimacteric near-isogenic line (NIL) SC7-2, and with the climacteric behavior of the NIL SC3-5-1, using their nonclimacteric inbred parentals, "Piel de Sapo" (PS) and PI 161375 (SC). PS was firmer and had a higher ripening index and greater hemicellulosic content than SC, with its lower wall material yield, and uronic acid, neutral sugar, cellulose and free sugar content and higher pectic content. SC3-5-1 showed lower uronic acid values, a higher soluble solid content, and similar flesh firmness to PS. SC3-5-1 yielded mainly high molecular weight polysaccharides in the imidazole-soluble fraction than PS. SC7-2 showed greater flesh firmness, a higher neutral sugar (especially galactose and mannose) and uronic acid content, together with a larger cellulose and α-cellulose residue than PS. SC7-2 also contained more polysaccharides of low molecular weight in the first pectic fraction and shifted toward higher molecular weights in the main peak of the 4 M potassium-soluble fraction compared with PS.     

Isolation and identification of phase II enzyme inductors obtained from black Shawaya sorghum [Sorghum bicolor (L.) Moench] bran

The antioxidant capacity as oxygen radical absorbance capacity (ORAC) and in vitro chemopreventive effect of phytochemicals extracted, partitioned and isolated from the decorticated bran of black Shawaya sorghum (Sorghum bicolor L. Moench) was studied. The chemopreventive effect was evaluated as the induction effect of quinone reductase (QR) using murine hepatoma cells Hepa 1c1c7. The crude methanol extract with an antioxidant capacity of 3.7 μmol Trolox equivalent (TE)/mg was partitioned into 120 fractions with fast centrifugal partition chromatography (FCPC). Fractions with partition coefficients between 0.1 and 0.26 exerted more than 50% of the antioxidant activity of the original crude extract and contained the highest concentration of anthocyanins. No correlation was found for ferulic acid content and ORAC or chemoprevention. A derivative of 7-methoxyapigeninidin at a concentration of 1 μg/ml increased by 31% the QR activity while the original extract reached a maximum of 27% at 40 μg/ml.     

Comparative pharmacokinetics and a clinical laboratory evaluation of intravenous acetaminophen in Beagle and Galgo Español dogs

Objective

To assess the pharmacokinetics (PK) and conduct a clinical laboratory evaluation of acetaminophen in Beagle and Galgo Español (GE) dogs.