Segmental aortic wall stiffness from intravascular ultrasound at normal and subnormal aortic pressure in pigs.

Segmental aortic wall stiffness was calculated from intravascular ultrasound images and intravascular pressures in six pigs at normal and subnormal aortic pressures (21 sequences of pressures and areas before and after boli of intravenous nitroglycerin). The wall stiffness was expressed as the pressure-strain elastic modulus (Ep). The Ep was calculated from the formula: Ep = delta PR delta R-1 (P, pressure; R, radius) in two different ways. First from maximal and minimal values of pressure and area. Second as the slope of linear regression line of delta PR as a function of delta R from 29 simultaneous recorded pressures and images. The average Ep value for all sequences in the different segments was 0.58 +/- 0.55 10(5) Pa (Method 1) and 0.50 +/- 0.40 10(5) Pa (Method 2). Ep increased with the distance from the heart at normal aortic pressures. At subnormal aortic pressures after intravenous nitroglycerin this relationship was not so evident. At subnormal aortic pressures the calculated Ep values were significantly reduced in the lower half of the abdominal aorta. The phase lag, i.e. hysteresis, between pressure and diameter was demonstrated. Our study shows the applicability of intravascular ultrasound as a tool to evaluate arterial wall stiffness.     

DNA immunization efficiently targets conserved functional domains of protease and ATPase/helicase of nonstructural 3 protein (NS3) of human hepatitis C virus

Nonstructural protein 3 (NS3) of human hepatitis C virus (HCV) is a conserved multi-functional protein essential for replication and translation of viral RNA and polyprotein processing. Early T-cell response against NS3 is capable of restricting viremia. We aimed at characterizing the immunogenicity in gene immunization of the conserved regions of NS3 critical for protein folding and activity. C57BL/6 mice were injected with NS3 gene of Russian HCV 1b isolate 274933RU. Immunization did not exert any overt histological changes and had no long-term effects on the immune status of NS3 gene-recipients. The immune response in NS3 gene-recipients was screened by antibody ELISA, T-cell proliferation test and immune assays for specific cytokine production. T-lymphocytes of NS3 gene-recipients proliferated in response to peptides representing conserved regions of protease and ATPase/helicase. Stimulated T-lymphocytes produced IL-2, and in response to protease-derived peptides, also IFN-gamma. Potent and long-lasting antibody response was raised against conserved NS3 regions including "Greek-key" motif of protease, motifs II, V and polynucleotide-binding domains of ATPase/helicase. Thus, gene immunization effectively targeted conserved regions critical for NS3 protease and helicase function. In type and specificity, immune response of NS3 gene-immunized mice mimicked immunity achieved in the acute self-limiting HCV infection of human and primates and in virus-exposed healthy individuals, indicating promiscuity of NS3 as immunogen.     

Partial purification and characterization of anhydrotetracycline oxygenase of Streptomyces aureofaciens

Anhydrotetracycline oxygenase was purified both by affinity chromatography and by hydrophobic interaction chromatography. Molecular weight of anhydrotetracycline oxygenase was determined to be 115,000 by Sephadex G-200 gel filtration. Using preparative isoelectric focusing the isoelectric point of the enzyme was estimated to be 5.3. The enzyme showed a sensitivity to thiol-specific inhibitors. During the hydrophobic interaction purification step, the activity dropped considerably. Reactivation occurred when a heat treated crude extract was added to the reaction mixture.     

Evaluation of psychomotor agility during the period of withdrawal symptom dissipation in patients with alcohol dependence type 1 and 2 or Cloninger's

28 alcohol dependent men were examined three times (on the 1st, 3rd or 4th and 7th day of withdrawal). The results confirmed the reported differences in the course of withdrawal syndrome in type 1 and type 2 alcoholics. Patients with type 2 alcoholism had more pronounced psychophysiological and cognitive disturbances. Tremor was more intensive in these patients and their reaction time was slower. Also, impaired estimation of passing time lasted longer, but at the same time their mood improved faster with the diminished intensity of withdrawal symptoms than in patients with type 1 alcoholism. The results confirm the possibility of diverse etiology in alcohol dependence.     

Inhibition of colony formation in agarose of metastatic human breast carcinoma and melanoma cells by synthetic glycoamine analogs

We studied the influence of 10 synthetic glycoamine analogs on colony formation in 0.3 and 0.9% agarose by metastatic human breast carcinoma (MDA-MB-435) and melanoma (TXM-13) cells. Nine synthetic analogs significantly inhibited the colony formation in 0.9% agarose of MDA-MB-435 human breast carcinoma cells; five compounds caused a 73–83% reduction of colony formation. Seven synthetic glycoamines caused a significant inhibition of colony formation in 0.9% agarose by TXM-13 melanoma cells with the inhibitory effect ranging from 71 to 87%. The 50% inhibition (I₅₀) doses and relative activity rank of the compounds were similar for both breast carcinoma and melanoma cell lines. The murine B16 melanoma cell aggregation assay was employed to elucidate the potential mechanism(s) of the inhibitory activity of synthetic glycoamines. The relative activity ranks of the compounds based on the independently determined I₅₀ doses for both cell aggregation and clonogenic growth assays were very similar for the four most active synthetic analogs and clearly indicated the importance of hydrophobic amino acid in mediating the bioactivity of synthetic glycoamines. In both experimental systems (clonogenic growth in agarose and cell aggregation assay) the leading compound was N-(1-deoxy-d-fructos-1-y1)-d-leucine (Fru-d-Leu) and the least active analog was N-(1-deoxy-d-fructos-1-yl)-glycine (Fru-Gly). These results show that synthetic glycoamines may act by competing for specific carbohydrate-lectin interactions, particularly those involving -galactoside-specific lectins expressed on metastatic cells.     

Implications of imidazolines and imidazoline receptors role at the vascular level

There are both post- and pre-synaptic vascular imidazoline (IM) binding sites. The importance of direct IM actions and that of peripheral imidazoline receptors (IRs) are shadowed by the central effects of IMs and by their interaction with alpha 2 adrenoceptors. Since the discovery of clonidine the many studies on IMs have been focused on their hypotensive effect, with rilmenidine and moxonidine as representative drugs. Formerly called IM preferring alpha 2 or IM/guanidium sites, the IRs (idazoxan-sensitive) are the plasmalemmal I1 (clonidine-sensitive) and the various I2 (one structure identified as MAO). I1 signaling includes activation of phosphatidylcholine-selective phospholipase C and inhibition of some ligand-gated channels. Inhibitory IRs on postganglionic sympathetic terminals, are not alpha 2, H3, I1 or I2. Some IMs directly affect CaL, while others inhibit K+ efflux. Clonidine-displacing substances including agmatine are endogenous ligands at IRs and alpha 2 and may participate in arterial pressure control. Beside few speculations, the roles of vascular IRs are largely unknown.     

Naja melanoleuca cobra venom contains two forms of complement-depleting factor (CVF)

Two forms of complement-depleting cobra venom factor (CVFm1 and CVFm2), possessing molecular masses of 142.6 kDa (CVFm1) and 143.1 kDa (CVFm2), according to MALDI mass-spectrometry, were isolated from the Naja melanoleuca cobra venom. As shown by polyacrylamide gel electrophoresis in the presence of SDS, both forms similarly to factor from the Naja kaouthia cobra venom (CVFk) consist of three polypeptide chains with molecular masses of about 70, 50, and 30 kDa, the two large subunits being glycosylated. As determined by MALDI mass-spectrometry, 30 kDa subunits of CVFm1 and CVFm2 have considerably different finger-prints of tryptic digests that suggests differences in their amino acid sequences. A study of activity in vivo has shown no significant differences in C3 consumption by CVFm1, CVFm2 and CVFk in mouse blood. However, as shown by an immunoassay method, they differ in their ability to activate the complement system via C3 conversion, the ratio of these activities for CVFm1:CVFm2:CVFk being 2.5:1.6:1. Kinetic studies using a hemolytic test showed that complement depletion by CVFm1 is faster than that by CVFm2. Thus, for the first time the presence in a single venom of two forms of CVF differing by both amino acid sequence and biological activity has been shown.