Expression of lysyl oxidase from cDNA constructs in mammalian cells: the propeptide region is not essential to the folding and secretion of the functional enzyme

Rat aortic lysyl oxidase cDNA was expressed under a metallothionein promoter in Chinese hamster ovary cells using a dihydrofolate reductase selection marker. One methotrexate-resistant cell line, LOD-06, generated by transfecting with full-length cDNA, yielded lysyl oxidase proteins consistent with the 50 kDa proenzyme and a 29 kDa mature catalyst. A second cell line, LOD32-2, was generated by transfection with a truncated cDNA lacking sequences which code for the bulk of the propeptide region. Both cell lines secreted apparently identical, 29 kDa forms of mature lysyl oxidase each of which catalyzed the deamination of human recombinant tropoelastin and alkylamines, consistent with the known specificity of lysyl oxidase. The secreted enzyme forms were inhibited by chemical inhibitors of lysyl oxidase activity, including beta-aminopropionitrile, phenylhydrazine, ethylenediamine, alpha, alpha'-dipyridyl, and diethyldithiocarbamate. Sensitivity to these agents is consistent with the presence of copper and carbonyl cofactors in the expressed enzymes, characteristic of lysyl oxidase from connective tissues. These results indicate the lack of essentiality of the deleted proprotein sequence for the proper folding, generation of catalytic function, and secretion of lysyl oxidase.     

Impact of minority physicians on health care

BACKGROUND: The fact that few doctors have their own GP may help explain doctors' poor health outcomes. Barriers to the doctor as patient role may arise from problematic perceptions of the 'doctor' subject position and from its subjugation of 'other-than-doctor' positions. Medical culture appears to foster and construct inappropriate health seeking behaviour of doctors. OBJECTIVE: A behaviour model is proposed--delusion, denial and delay; self-investigation, self-diagnosis, self-treatment and self-referral. Application of a behaviour change model could facilitate the doctor as patient role. DISCUSSION: Literary theory and sociology critiques may help peel off the social construction of 'doctor'. The realisation of such myths may allow access to medical care enjoyed by the general community.     

Thoracic MR aortography: imaging techniques and strategies

Three-dimensional (3D) gadolinium-enhanced magnetic resonance (MR) angiography is a promising technique for thoracic aortography that complements electrocardiographically gated T1-weighted spin-echo imaging and cine MR imaging. Axial and left anterior oblique T1-weighted spin-echo images are well suited to measurement of aortic dimensions and evaluation of aortic aneurysms. Sagittal and coronal spin-echo images are helpful in evaluation of vascular rings and aortic dissection. Cine gradient-echo and cine phase-contrast imaging allow dynamic evaluation of aortic and valvular flow. Cine phase-contrast imaging also enables noninvasive quantification of blood flow. Capable of being performed during a single breath hold, 3D gadolinium-enhanced MR angiography provides high-resolution 3D data that can be readily used for projection angiography and multiplanar reformation. This technique enables further demonstration of subtle pathologic conditions. Three-dimensional gadolinium-enhanced MR angiography allows more comprehensive and efficient evaluation of the thoracic aorta.     

Dynamics of the marital structure with respect to the nationality and birthplace of spouses in a modern urbanized Siberian population (as exemplified by Tomsk)

To analyze a population's marital structure with respect to the ethnicity and birthplace of the spouses and to estimate the indices of endogamy, migration, and marriage assortativeness, the records of marriages in Tomsk during two periods of time (1970-1972 and 1985-1990) were studied. The parameters of the population-genetic and demographic structure proved to change during the studied period: the endogamy index and the indices of marriage assortativeness increased, and the migration index decreased. These data suggest that the genetic structure of the Tomsk urbanized population stabilized with time and that the intrapopulation subdivision decreased (a tendency for panmixia).     

Functional stereotaxy with magnetic resonance tomographic guidance

The paper summarizes experience with stereotactic operations by magnetic resonance imaging data by using a Russian OREOL stereotactic manipulator. It describes methods of preoperative preparation of patients by employing diagnostic magnetic resonance tomographs. A number of points of the procedure for identification and localization of deep target structures from magnetic resonance images in parkinsonism, temporal epilepsy and some mental disorders.     

Brain biopotential mapping in emotional and cognitive pathology

The EEG rhythms spectral power and intracortical connections were studied in depressive and schizophrenic patients in four schematically outlined main cortical quadrants. In depressive patients two cortical quadrants--right anterior and left posterior ones appeared to be predominantly involved in negative emotions regulation. Alpha-rhythm spectral power and cortical connections in these areas were decreased, pointing to their relative hyperactivation and functional disturbances. Schizophrenics patients were divided into two groups--with the predominance of "positive" and "negative" symptoms. In the first group there was revealed the decrease of the most EEG rhythms spectral power only in parietal areas, on the second one--in all cortical areas. There was also the significant alpha-rhythm spectral power asymmetry, different in the two groups of schizophrenic patients. In "positive" schizophrenics the decrease of alpha-rhythm spectral power was revealed in parietal and occipital areas of the right hemisphere, and in "negative" ones--vice versa, these results pointing of the relatively higher activation of the right posterior quadrant in the former group and of the left posterior quadrant in the latter one. In the anterior cortical quadrants the intracortical connections were however higher in the left hemisphere in "positive" and in the right one--in "negative" schizophrenics. Thus, there are some contradictory results obtained by different methods in anterior and posterior cortical quadrants in each hemisphere, these results being of opposite directions in patients with "positive" and "negative" manifestations. "Region of interest" (ROI) in schizophrenics appeared to be predominantly the frontal and temporal ones, while the "Rhythm of interest"--beta2.     

The effect of activated macrophage products on neurite growth in an organotypic culture of chick embryo sensory ganglia]

Murine peritoneal macrophages, activated by BCG vaccine, and human peripheral blood monocytes, activated by lipopolysaccharides, exerted neurite stimulating or neurite inhibiting effects in various periods of activation. The supernatants of these preparations were active in organotypic culture of chick embryo dorsal root ganglia. The inhibition of neurite growth on the 1st day of cultivation was followed by the neurite-stimulating effect. The fluctuation of neurite-inhibition and neurite-stimulation effect of macrophage supernatants suggest the availability of certain changes in cytokine composition in different periods of macrophage activation.     

Functional analysis of the DNA-packaging/terminase protein gp17 from bacteriophage T4

In bacteriophage T4, the terminase complex constituted by the large subunit gp17 (69 kDa) and the small subunit gp16 (18 kDa) is a critical component of the ATP-driven DNA-packaging pump that translocates DNA into an empty capsid shell. Evidence suggests that the large subunit gp17 is the critical component and consists of a number of the functional sites required for DNA-packaging. It exhibits a terminase activity that introduces non-specific cuts into DNA, a portal vertex binding site that allows linkage of cleaved DNA to an empty prohead, an in vitro DNA-packaging activity, and an ATPase activity. In addition, a consensus metal-binding motif and two consensus ATP-binding sites have been identified by sequence analysis. In order to understand the mechanism of action of the multifunctional gp17, we developed an expression-based selection strategy to select for mutants that are defective in terminase function. Characterization of one of the mutants revealed a unique phenotype in which a single H436R mutation resulted in a dramatic loss of both the terminase and the DNA-packaging functions. Indeed, in vivo substitution of H436 with any of the 12 amino acids for which a suppressor is available was lethal to T4 development. According to one hypothesis, H436 is part of a metal-binding motif that is essential for gp17 function. This hypothesis was tested by introducing mutations at each of the three histidine pairs, the H382-X2-H385 pair, the H411-X2-H414 pair and the H430-X5-H436 pair, which constitute the histidine-rich region near the C terminus of gp17. A mutation at either the H411 pair or the H430 pair resulted in a loss of gp17 function, whereas a mutation at the H382 pair had no effect. In addition to the putative metal-binding motif, substitutions at residue K166 within the putative N terminus-proximal ATP-binding site also resulted in a loss of gp17 function. We propose that a metal-binding motif involving the histidine residues within the sequence H411-X2-H414-X15-H430-X5-H436 is essential for gp17 function. Metal-terminase interactions may be required for structural alignment and stabilization of functional sites in phage T4 terminase and other double-stranded DNA phage terminases.     

Prospects for development of functional diagnosis in antituberculosis institutions in the Russian Federation

The proposed method for measuring hyaluronidase activity of microorganism is based on prevention of hyaluronic acid clot formation with rivanol under the effect of hyaluronidase. This made possible the quantitative and qualitative detection of hyaluronidase activities of different staphylococcus species and strains. The maximum level of the enzyme and highest rate of its detection were typical of St. aureus. Its strains producing hyaluronidase in quantities of at least 0.5 IU are significantly (p < 0.01) more often isolated from medical staff.     

Various combinations of antihypertensive drugs: comparative assessment in elderly patients]

Our objective in this study was a comparative evaluation of efficiency of modern drug alternatives to treat peptic ulcer, based on new algorithms of diagnosis and treatment. The results obtained suggest to us that in a major proportion of cases uncomplicated duodenal ulcers might be efficiently managed in an outpatient-clinic setting even with a two-to-four week monotherapy. The best results of treatment are recordable in those groups of patients having received inhibitors of proton pump.