Functional characteristics of rat gastrocnemius and tibialis anterior muscles during growth

Several morphological and functional characteristics of the rat gastrocnemius medialis and tibialis anterior muscle were studied in young, adult, and old rats to assess the influence of growth. Antagonist muscles were studied to determine how changes of muscle architecture and functional characteristics are influenced by the demands of increased body weight and by the specific roles of these muscles in locomotion. Both muscles change drastically, for instance, in muscle length, volume, physiological cross-sectional area aponeurosis length, and their muscular geometry changes allometrically for both muscles. The relationships between muscle length, distance between origin and insertion, tendon length, and tibial length also change with growth. Both muscles are rather pennate, so that the increase of physiological cross-sectional area is a major factor in the determination of muscle length. No significant difference could be shown for fundamental physiological characteristics (i.e., functional characteristics normalized for muscular dimensions such as maximal work per unit volume). The changes of morphological and functional variables of both muscles parallel each other as is apparent from the index of antagonist characteristics, which is constant for all variables studied with the exception of muscle volume and tendon length. Consequently, the considerable and similar changes of TA and GM morphology and functional characteristics that take place during growth from approximately four weeks postnatally is not caused by changes of muscular material but by changes of the amount and architectural arrangement of the material involved.     

Online solid phase extraction with liquid chromatography–tandem mass spectrometry to analyze remoxipride in small plasma-, brain homogenate-, and brain microdialysate samples

Remoxipride is a selective dopamine D₂ receptor antagonist, and useful as a model compound in mechanism-based pharmacological investigations. To that end, studies in small animals with serial sampling over time are needed. For these small volume samples currently no suitable analytical methods are available. We propose analytical methods for the detection of low concentrations remoxipride in small sample volumes of plasma, brain homogenate, and brain microdialysate, using online solid phase extraction with liquid chromatography–tandem mass spectrometry. Method development, optimization and validation are described in terms of calibration curves, extraction yield, lower limit of quantification (LLOQ), precision, accuracy, inter-day- and intra-day variability. The 20 μl plasma samples showed an extraction yield of 76%, with a LLOQ of 0.5 ng/ml. For 0.6 ml brain homogenate samples the extraction yield was 45%, with a LLOQ of 1.8 ng/ml. The 20 μl brain microdialysate samples, without pre-treatment, had a LLOQ of 0.25 ng/ml. The precision and accuracy were well within the acceptable 15% range. Considering the small sample volumes, the high sensitivity and good reproducibility, the analytical methods are suitable for analyzing small sample volumes with low remoxipride concentrations.     

The presence of gap junctions during early Patella embryogenesis: An electron microscopical study

The presence of gap junctions has been examined up to the sixth cleavage in the early Patella embryo. Gap junctions are located all over the blastomere borders. In 2-, 4-, and 8-cell embryos they were also observed at peripheral contacts. The frequency and size of the gap junctions increase at the 32-cell stage. The structure of gap junctions is similar in all stages investigated with hexagonally arranged equal-sized particles (11 nm) having a constant center-to-center spacing (13.0 nm). At the 32-cell stage formation plaques were observed, indicating an increase of gap junctions.     

Enhanced production of human gamma-interferon in nylon column-fractionated cell cultures

The production of human gamma-interferon (HuIFN-gamma) in unfractionated and nylon wool column-fractionated leukocyte cell cultures stimulated with PMA and PHA was investigated. Production was studied with normal and reduced autologous serum protein levels in 96-hr spinner cultures. A 10- to 15-fold enhancement of production and a 50-fold increase in specific activity of crude HuIFN-gamma was demonstrated in nylon column-fractionated/reduced serum cell cultures. Kinetic analysis revealed a production rate maximum within 6 hr of induction in unprocessed cell cultures, whereas production occurred at an essentially constant rate for 48 hr in fractionated cell cultures.     

Interaction of methyl-xanthines with myeloperoxidase. An anti-inflammatory mechanism.

1. Inhibition of myeloperoxidase (MPO)-catalyzed reactions by methyl-substituted xanthines has been investigated. 2. Except for theobromine and caffeine, all xanthines tested were potent inhibitors of the MPO-H2O2-Cl- system. 3. In contrast to methyl substitution in the 1 or 8 position of xanthine, substitution in the 3 or 7 position had a marked effect on the inhibition of MPO catalysis. 4. Two different inhibitory mechanisms were induced; scavenging of hypochlorous acid (HOCl) generated by the MPO system and accumulation of Compound II (ferryl MPO) which is inactive as a catalyst of Cl- oxidation.     

Antimycin binds to a small subunit of the ubiquinol: cytochrome c oxidoreductase

Bovine heart ubiquinol: cytochrome c oxidoreductase in Triton X-100 is split with guanidine into a number of fractions. A new method for measuring antimycin binding is developed using extraction with pentanol of the reversibly bound antimycin. By this method and the normal titration method, antimycin-binding capacity is found in a fraction containing a small subunit with a molecular mass of about 12000. This polypeptide was associated with cytochrome c1 but is probably not the 'hinge protein'. Fractions that contain cytochrome b did not show binding by the pentanol-extraction method.     

Lymphatic drainage in patients after replantation of extremities

Lymph drainage was studied by means of lymph scintigraphy in eight patients in whom successful replantation of a totally or subtotally amputated extremity had been performed. Scintigrams were made after subcutaneous injection of technetium-99m in the replanted part of the patient and the contralateral, normal extremity. In all scintigrams, axillary or inguinal lymph node activity is seen, implying drainage of lymph by means of the lymph vessels. Retention of colloid in the replanted part (79 to 94 percent) shows no significant difference with the contralateral, normal side (86 to 94 percent). Unquestionable evidence of regeneration of lymphatics in humans is delivered in the three patients, in whom lymph node activity and normal retention percentages are seen on the scintigrams after total amputation of an extremity followed by replantation without anastomosing of interrupted lymph vessels.     

Biorefining of wood: combined production of ethanol and xylanase from waste fiber sludge

The possibility to utilize fiber sludge, waste fibers from pulp mills and lignocellulose-based biorefineries, for combined production of liquid biofuel and biocatalysts was investigated. Without pretreatment, fiber sludge was hydrolyzed enzymatically to monosaccharides, mainly glucose and xylose. In the first of two sequential fermentation steps, the fiber sludge hydrolysate was fermented to cellulosic ethanol with the yeast Saccharomyces cerevisiae. Although the final ethanol yields were similar, the ethanol productivity after 9.5?h was 3.3?g/l/h for the fiber sludge hydrolysate compared with only 2.2?g/l/h for a reference fermentation with similar sugar content. In the second fermentation step, the spent fiber sludge hydrolysate (the stillage obtained after distillation) was used as growth medium for recombinant Aspergillus niger expressing the xylanase-encoding Trichoderma reesei (Hypocrea jecorina) xyn2 gene. The xylanase activity obtained with the spent fiber sludge hydrolysate (8,500?nkat/ml) was higher than that obtained in a standard medium with similar monosaccharide content (1,400?nkat/ml). Analyses based on deglycosylation with N-glycosidase?F suggest that the main part of the recombinant xylanase was unglycosylated and had molecular mass of 20.7?kDa, while a minor part had N-linked glycosylation and molecular mass of 23.6?kDa. Chemical analyses of the growth medium showed that important carbon sources in the spent fiber sludge hydrolysate included xylose, small aliphatic acids, and oligosaccharides. The results show the potential of converting waste fiber sludge to liquid biofuel and enzymes as coproducts in lignocellulose-based biorefineries.