Carbohydrate molecular weight profiling, sequence, linkage, and branching data: ES-MS and CID

This section summarizes several strategies for a more complete understanding of carbohydrate structure with a focus on glycolipids and glycoprotein glycans. The techniques include periodate oxidation to impart greater molecular specificity to isomeric glycans, methylation to improve sensitivity and the information content within CID spectra, electrospray for "soft" and efficient ionization, and CID to obtain structural detail. The lipophilicity of the products following derivatization contributes to product cleanup by solvent extraction and enhances sensitivity during ES. When combined with CID information, this yields sequence, linkage, and branching information. Oxidation and reduction preceding methylation augments CID analysis with an altered structure that can be profiled at the same sensitivity. Within the context of established motifs, these contrasting profiles corroborate glycan structure and specifically identify isobaric elements transparent in the initial profile. An earlier report indicating ring-opening fragments were essentially absent in low-energy collisions of methylated and natriated oligosaccharides contrasts our observations. However, as this report used a methylated oligomer containing an internal N-acetylhexose as an illustration, the conclusion is plausible (cf., Figure 9). The poor ionization efficiency of FAB and the high matrix background limit the dynamic range in the CID spectrum and, thereby, the ability to unambiguously identify weaker peaks. It would be expected that high-energy CID affords a broader range of fragment types, including ring-opening fragments. In terms of a structural methodology, this is ambivalent since the increase in fragmentation pathways also applies to small molecule eliminations which are usually less informative. In ES-CID-MS, the carbohydrate chemist has a powerful new tool in hand for structural elucidations that can be conducted at the low-picomole level. Parallel developments can be expected to continue for other ionization methods, in particular matrix-assisted desorption/ionization on linear and reflectron time of flight mass spectrometers, and improvement in the performance and sensitivity of high-resolution mass analyzers through the use of focal plane detectors and more sophisticated hardware and software for Fourier transform ion cyclotron resonance mass measurements. These have, as yet, only begun to be applied to carbohydrate structural analysis but should add still more versatility to experimental design in the future.     

Low- and high-density lipoproteins as mitogenic factors for vascular smooth muscle cells: individual, additive and synergistic effects

The mitogenic activities of low (LDL)- and high (HDL)-density lipoproteins have been examined in cultures of human vascular smooth muscle cells (VSMC). LDL and HDL3 dose-dependently (EC50 values approximately 50 micrograms/ml) stimulated DNA and protein synthesis ([3H]-thymidine and [3H]-leucine incorporation, respectively) in the absence of exogenously added mitogens. The synthetic responses of VSMC to combinations of LDL and HDL3 were additive, indicating that each lipoprotein mediates discrete effects. LDL or HDL3 promoted VSMC proliferation under strict mitogen-free conditions, but this growth response was not sustained. VSMC exposed to combinations of lipoproteins (either LDL or HDL3) and growth factors (either PDGF-BB, EGF, bFGF or IGF) exhibited synergistic DNA synthesis responses. In the combined presence of PDGF-BB and either LDL or HDL3, VSMC proliferation was sustained. Anionized lipoprotein preparations (oxidized, acetylated, carbamylated or malonimylated) also stimulated DNA and protein synthesis. Since the antioxidant beta-hydroxylated toluene did not block the effect of native LDL on DNA synthesis, and fucoidin, a specific competitor for the 'scavenger' receptor, did not inhibit oxidized LDL-induced DNA synthesis, activation of mitogenic signals by lipoproteins does not depend on lipid peroxidation. Rather, the apparent intrinsic mitogenic potential of lipoproteins may depend upon their direct activation of replication-coupled signal transduction systems.     

Activity of threonine deaminase and biosynthesis of avermectins in the culture of Streptomyces avermitilis

The influence of ammonium, threonine, isoleucine and valine on the activity of threonine deaminase and the biosynthesis of avermectins in the culture of two mutants of Streptomyces avermitilis, i.e. a sensitive one and a resistant one with respect to alpha-amino-beta-oxyvaleric acid, a threonine antimetabolite, was studied. It was shown that the synthesis of threonine deaminase was induced by threonine and valine in the mycelium of both the mutants. The level of threonine deaminase was higher in the mycelium of the antimetabolite resistant mutant. The antibiotic activity of the resistant mutant was lower while the relative content of the group B avermectins in the pool of the synthesized avermectins was higher than that in the culture of the sensitive mutant.     

Essential phospholipids in the treatment of chronic pyelonephritis]

Through TCD test, observation on the blood flow dynamics change before and after treatment with 31 cases of the child cerebral atrophy was made. It is found that scalp therapy is able to speed up the blood flow of part artery, especially increase the even blood flow speed of MCA, ACA obviously (P < 0.05), which preliminary shows the mechanism of scalp therapy for child cerebral atrophy mean while. It is found that scalp therapy also has the function to restrain epilepsy.     

Making of the clinicophysiological diagnosis within the system of medical flight certification

Adaptation responses were evaluated in flying personnel during medical examination with the use of simulators of the main dynamic factors of flight. Utility of assessing the adaptive reactions of the cardiovascular system by their physiological "cost" has been demonstrated. Parameters of the normal cardiovascular response to moderate hypoxia in altitude chamber have been established. On the basis of accumulated data, more sensitive and informative criteria for assessment of the functional state of the human organism are proposed for clinicophysiological diagnostics within the system of medical certification of flying personnel.     

Re-entrant activity and its control in a model of mammalian ventricular tissue

We characterize the meander of re-entrant excitation in a model of a sheet of mammalian ventricular tissue, and its control by resonant drift under feedback driven stimulation. The Oxsoft equations for excitability in a guinea pig single ventricular cell were incorporated in a two dimensional reaction-diffusion system to model homogeneous, isotropic tissue with a plane wave conduction velocity of 0.35 m s-1. Re-entrant spiral wave solutions have a spatially extended transient motion (linear core) that settles down into rotation with an irregular period of 100-110 ms around an irregular, multi-lobed spiky core. In anisotropic tissue this would appear as a linear conduction block. The typical velocity of drift of the spiral wave induced by low amplitude resonant forcing is 0.4 cm s-1.     

Use of a DNA polymerase III bypass mutant of Escherichia coli, pcbA1, to isolate potentially useful mutations of a complex plasmid replicon

We report a case of a 63-year-old woman who presented with pseudoaneurysm of the free wall of the left ventricle secondary to myocardial infarction, in the presence of angiographically normal major coronary arteries. This is the only such case we know of, in which the patient underwent successful surgical correction. At last follow-up, the patient was in good condition with no evidence of cardiac disease, at 9 years after surgery.     

Serial cytological assay of micronucleus induction: a new tool to predict human cancer radiosensitivity

The role of the propeptide sequence and a disulfide bridge between sites 1 and 122 in chymotrypsin has been examined by comparing enzyme activities of wild-type and mutant enzymes. The kinetic constants of mutants devoid of the Cys1-Cys122 disulfide-linked propeptide show that this linkage is not important either for activity or substrate specificity. However this linkage appears to be the major factor in keeping the zymogen stable against non-specific activation. A comparison of zymogen stabilities showed that the trypsinogen propeptide is ten times more effective than the chymotrypsinogen propeptide in preventing non-specific zymogen activation during heterologous expression and secretion from yeast. This feature can also be transferred in trans to chymotrypsinogen; i.e. the chymotrypsin trypsin propeptide chimera forms a stable zymogen.