Chondroitin sulphate at the endothelial lumen of pig aorta: Assay by radiolabelling and by X-ray microanalysis

Trypsin-releasable glycosaminoglycans from the luminal surface of intact pig aorta were measured following metabolic labelling with³⁵S]sulphate. Chondroitin sulphate was found to be present at a surface density equal to that already established for heparan sulphate (5×10¹¹ chains per cm²). This result was confirmed by X-ray microanalysis of the luminal sulphur content before and after treatment with specific glycosaminoglycan-degrading enzymes. This result implies that approximately half of the luminal surface is occupied by sulphated glycosaminoglycans.     

The use of 13C-n.m.r. spectroscopy to monitor alginate biosynthesis in mucoid Pseudomonas aeruginosa.

The biosynthesis of alginate by a mucoid strain of Pseudomonas aeruginosa, isolated from a cystic-fibrosis patient, was monitored by using 13C-n.m.r. spectroscopy of bacterial cultures incubated with 1-13C- or 2-13C-enriched fructose. When 1-13C- or 2-13C-enriched fructose was used as the precursor of alginate, enrichment with 13C in the constituent uronic acid monomers of the polysaccharide could only be detected in C-1 or C-2 respectively, indicating that alginate is synthesized in Ps. aeruginosa directly from fructose, with the hexose molecule being retained intact; this rules out the involvement of C3 intermediates, which occurs when glucose is the alginate precursor. The absence of detectable poly-L-gluluronate block sequences from the alginate of Ps. aeruginosa was confirmed, and it was shown that there is no modification of the arrangement of the constituent uronic acids between polymerization to form alginate and the appearance of the mature alginate in the extracellular medium. The 13C-n.m.r. data also provided independent evidence for acetylation on D-mannuronate residues and for the ratio of D-mannuronate to L-guluronate residues in newly synthesized alginate, which had previously been determined only for material secreted from bacteria into the extracellular medium.     

Modification of functional arginine residues in purified bovine testicular hyaluronidase with butane-2, 3-dione

The effect of mononucleotides on the cytosolic rat liver glucocorticoid receptor has been studied by the use of aqueous dextran-poly(ethylene glycol) two-phase partitioning. During incubations in 0.4 M KCl at 0 degrees C, millimolar concentrations of ADP and ATP, but not AMP, CTP, UTP and GTP, inhibit the increase in the receptor partition coefficient associated with receptor activation. This inhibition is counteracted by millimolar concentrations of theophylline and MgCl2. Two nonhydrolyzable analogues of ATP, alpha, beta-Methyleneadenosine 5'-triphosphate and beta, gamma-methyleneadenosine 5'-triphosphate, also inhibit the increase in the partition coefficient. alpha, beta-Methyleneadenosine 5'-triphosphate is much more potent than ATP in doing so, and this compound was also shown to reduce the amount of receptor to bind to DNA-Sepharose after the incubations. Thus, adenine nucleotides induce a change in the state of the receptor, apparently consisting in an inhibition of receptor activation.     

Isolation,characterisation and expression of a cDNA for pea cholinephosphate cytidylyltransferase

In plants, phosphatidylcholine is the major phospholipid in extra-plastid membranes and is synthesised mainly by the CDP-choline pathway. Evidence from studies in animals, as well as in plants, suggests that the intermediate step catalysed by cholinephosphate cytidylyltransferase (CPCT) has a major control in carbon flux to this lipid. We have isolated a full-length CPCT cDNA (designated PCT2) from Pisum sativum cv. Feltham First using an Arabidopsis probe and the polymerase chain reaction (PCR). The deduced amino acid of PCT2 is 48%, 43% and 76% identical to the rat, yeast and Brassica napus amino acid sequences, respectively. Expression of the CPCT protein in Escherichia coli confirmed the activity of the enzyme. Expression of the PCT2 mRNA in pea roots and stems was increased by treatment with 0.1 µM indole-3-acetic acid.     

The long-term genetic stability and individual specificity of the human gut microbiome

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Non-linear dynamics in muscle fatigue and strength model during maximal self-perceived elbow extensors training

Our aim was to determine the dynamics in muscle strength increase and fatigue development during repetitive maximal contraction in specific maximal self-perceived elbow extensors training program. We will derive our functional model for m. triceps brachii in spirit of traditional Hill’s two-component muscular model and after fitting our data, develop a prediction tool for this specific training system. Thirty-six healthy young men (21±1.0 y, BMI 25.4±7.2 kg/m²), who did not take part in any formal resistance exercise regime, volunteered for this study. The training protocol was performed on the isoacceleration dynamometer, lasted for 12 weeks, with a frequency of five sessions per week. Each training session included five sets of 10 maximal contractions (elbow extensions) with a 1 min resting period between each set. The non-linear dynamic system model was used for fitting our data in conjunction with the Levenberg–Marquardt regression algorithm. As a proper dynamical system, our functional model of m. triceps brachii can be used for prediction and control. The model can be used for the predictions of muscular fatigue in a single series, the cumulative daily muscular fatigue and the muscular growth throughout the training process. In conclusion, the application of non-linear dynamics in this particular training model allows us to mathematically explain some functional changes in the skeletal muscle as a result of its adaptation to programmed physical activity—training.     

Plate Assay for Simultaneous Detection of Alginate Lyases and Determination of Substrate Specificity

A plate assay to detect the presence of alginate lyases (EC 4.2.2.3) has been developed. The simultaneous use of specific alginate block structures of defined composition allows the substrate specificity of the enzymes to be determined. Clearing zones in the alginate-containing media are visualized with either cetyl pyridinium chloride or ruthenium red.