Hemocompatibility in hemodialysis and erythropoietin therapy

Two studies designed to investigate the effect of recombinant human erythropoietin (rHuEPO) treatment of anemia in chronic dialysis patients on hemocompatibility were conducted. Study 1, whose main aim was to establish whether treatment with rHuEPO enhances coagulation activation during dialysis, included 15 patients before rHuEPO therapy at a mean hematocrit (HCT) of 22.3% and then during therapy at a HCT of 29.3%. The plasma concentrations of the thrombin-antithrombin III complex were not higher during rHuEPO therapy than before it when performing hemodialysis with a Cuprophan membrane. No significant difference was demonstrated either in the values of activated clotting times (Hemochron), thrombocyte or white blood cell counts (Coulter S+II), or in plasma C5a concentrations (ELISA) established during dialysis sessions before and during rHuEPO therapy. In Study 2, which focused primarily on the question of whether or not rHuEPO therapy increases thrombocyte activation during hemodialysis, 8 patients on chronic dialysis were examined both before therapy at a mean HCT value of 22.1% and during rHuEPO therapy at a HCT of 31.5%, invariably during dialysis with either a Cuprophan or polyacrylonitrile (AN69HF) membrane. The plasma concentrations of beta-thromboglobulin (ELISA) did not differ between the examinations made during rHuEPO and before rHuEPO therapy; however, statistically significant differences were found between dialysis sessions involving Cuprophan and AN69HF membranes. No significant difference between examination before and during rHuEPO was demonstrated in activated clotting time nor thrombocyte and white blood cell counts in this study either. The authors conclude that rHuEPO therapy does not enhance coagulation activation during hemodialysis, does not have an effect on thrombocyte activation, and does not influence complement activation and changes in white blood cell counts.     

NADPH-diaphorase-stained neurons after experimental epilepsy in rats

The aim of this study was to determine the neuronal participation of nitric oxide (NO) in experimental epilepsy. To reach this objective, we established the amount of cells presenting nitric oxide synthase (NOS) and the amygdaline concentrations in the L-arginine-nitric oxide synthesis pathway. A group of fully epileptic rats, induced by the kindling procedure and that had reached at least 10 generalized seizures, was studied. We evaluated behavioral stages, electroencephalographic activities, and histochemical NOS-positive cells and carried out high-pressure liquid chromatography (HPLC) determinations of arginine, citrulline, and glutamic acid. Our results showed that behavioral and electrographic frequency, and duration of epileptic activities, were increased during the kindling process. Image processing system of NOS cells showed two types of intensities in cell stains in hippocampus, caudate-putamen, and amygdala. When we independently counted the two types of NOS stain cells, a selective increase in the number and density of weak-stained cells was observed, while dark-stained cells did not change in the studied structures. Additionally, arginine, citrulline, and glutamic acid concentrations in amygdala increased in kindled animals. The differential and specific increase in the stained cells expressing the nitric oxide synthase, as well as the increase in concentrations of the L-arginine-nitric oxide pathway in amygdala, suggested a relationship with the progressive augmentation in the electrophysiological hyperactivity characteristic of generalized epilepsy.     

Cellular UDP-glucose deficiency caused by a single point mutation in the UDP-glucose pyrophosphorylase gene

We previously isolated a mutant cell that is the only mammalian cell reported to have a persistently low level of UDP-glucose. In this work we obtained a spontaneous revertant whose UDP-glucose level lies between those found in the wild type and the mutant cell. The activity of UDP-glucose pyrophosphorylase (UDPG:PP), the enzyme that catalyzes the formation of UDP-glucose, was in the mutant 4% and in the revertant 56% of the activity found in the wild type cell. Sequence analysis of UDPG: PP cDNAs from the mutant cell showed one missense mutation, which changes amino acid residue 115 from glycine to aspartic acid. The substituted glycine is located within the largest stretch of strictly conserved residues among eukaryotic UDPG:PPs. The analysis of the cDNAs from the revertant cell indicated the presence of an equimolar mixture of the wild type and the mutated mRNAs, suggesting that the mutation has reverted in only one of the alleles. In summary, we demonstrate that the G115D substitution in the Chinese hamster UDPG:PP dramatically impairs its enzymatic activity, thereby causing cellular UDP-glucose deficiency.     

Effect of sulphate removal on the surface texture and acid-base properties of TiO2 (anatase)

Commercial anatase containing an appreciable amount of sulphur has been treated for sulphur removal by calcination at increasing temperatures or soaking with aqueous NaOH solutions. It has been found that both methods are active for sulphur removal, but the first one also leads to a sharp sintering of the titania particles. On the other hand, treatment with NaOH_aq leads to incorporation of sodium cations. Both treatments lead also to changes in surface acidity, as measured by Fourier transform infrared (FT-IR) monitoring of pyridine adsorption.     

Characterization of a lectin-like protein isolated from Lachesis muta snake venom

A lectin-like protein was isolated from L. muta venom by gel filtration on BIO Gel P-100 followed by column Chromatography on DEAE-sephades A-50. The protein eluted at 0.4 M Nacl in 0.01 Tris pH 7.3 and exhibited agglutinin activity toward 0+ human erythrocytes. The protein is a dimer with Mr 28 kDa. Amino acid analysis revealed high content of tryptophan and acid recidues and low content of cysteine and methionine residues. No neutral carbohydrates and sialic acid were detected. Circular dichroic spectrum shows 78% of B structure and 1% of alpha structure. In vitro experiments with erythrocytes from rat, rabbit and dog revealed strong agglutination while red blood cells from mice, sheep and goat were not agglutinated. In vivo experiments using anesthetized rats, a sharp and prolonged fall in the blood pressure was observed at protein dose of 1.5 mg/kg. Double dose of protein caused the death of the animal.     

High temperature ferroelastic phase in LiKSO4 crystals

Optical studies of LiKSO₄ crystals were performed in polarized 11yh.t within the temperature range from 300 up to 1000 K. The observations confirmed the existence of two phase transitions at about 700 and 940 K respectively. The high-temperature phase between 700 and 940 K was found to be ferroelastic.