Purification and regulation of glutamine synthetase in a collagenolytic Vibrio alginolyticus strain

Glutamine synthetase (EC 6.3.1.2) has been purified from a collagenolytic Vibrio alginolyticus strain. The apparent molecular weight of the glutamine synthetase subunit was approximately 62,000. This indicates a particle weight for the undissociated enzyme of 744,000, assuming the enzyme is the typical dodecamer. The glutamine synthetase enzyme had a sedimentation coefficient of 25.9 S and seems to be regulated by a denylylation and deadenylylation. The pH profiles assayed by the -glutamyltransferase method were similar for NH₄-shocked and unshocked cell extracts and isoactivity point was not obtained from these eurves. The optimum pH for purified and crude cell extracts was 7.9. Cell-free glutamine synthetase was inhibited by some amino acids and AMP. The transferase activity of glutamine synthetase from mid-exponential phase cells varied greatly depending on the sources of nitrogen or carbon in the growth medium. Glutamine synthetase level was regulated by nitrogen catabolite repression by (NH₄)₂SO₄ and glutamine, but cells grown, in the presence of proline, leucine, isoleucine, tryptophan, histidine, glutamic acid, glycine and arginine had enhanced levels of transferase activity. Glutamine synthetase was not subject to glucose, sucrose, fructose, glycerol or maltose catabolite repression and these sugars had the opposite effect and markedly enhanced glutamine synthetase activity.Abbreviations GS glutamine synthetase - SMM succinate minimal medium - ASMM ammonium/succinate minimal medium - GT -glutamyl transferase - SVP snake venom phosphodiesterase     

Reduction in headache patients' medical expenses associated with biofeedback and relaxation treatments

Comparisons are made of self-reported medical costs from a sample of headache patients who underwent various combinations of relaxation training and biofeedback training. The average costs for the 2 years prior to self-regulatory treatment were $955±480 (3 SEM) for 45 patients; for the 2 years after completing treatment the average costs were $52±28 (3 SEM) for patients. Within the limitations of the study, medical costs do seem to have been markedly reduced.This research was supported by a grant from NINCDS, NS-15235.     

Influenza vaccination in the elderly: 2. The economics of sending reminder letters

Reminder letters and follow-up telephone calls were used to increase influenza vaccination acceptance by 273 well elderly registered at an urban community health centre. The net effect of the reminder letters was to increase overall coverage to 43%, from 17% in the previous year. Follow-up telephone calls to patients who had not responded to the letters increased coverage to only 55%. Calculation of costs per additional vaccination given revealed that the use of reminder letters alone was much more cost-effective than follow-up telephone calls in increasing coverage. However, with the current fee-for-service reimbursement by medical care insurance in Ontario, neither means of improving vaccination coverage would result in net practice earnings. The implications for an effective and efficient annual influenza program in Canada are discussed.     

Physiological characteristics of the tympanic organ in noctuoid moths

Summary The tympanic organ ofSpodoptera frugiperda, Mocis latipes, Erebus odorata (Noctuidae) andMaenas jussiae (Arctiidae) was stimulated with acoustic stimuli of 20 kHz, 45 ms and 5 s duration, and intensities ranging from 30 to 100 dB. The electric activity of the auditory receptors was recorded at the tympanic nerve with a stainless steel hook electrode. In all of these moth species there is an intensity range (ca. 20 dB) in which the response of each auditory receptor (A₁ and A₂ cells) to 45 ms pulses varies in a linear relation to the logarithm of stimulus intensity. For intensities higher than this value, depending on the species and the cell analysed, the spike discharge may continue to increase, may saturate or may diminish (Fig. 2). InE. odorata andM. latipes the A₁-cell response shows a decrease for stimulus intensities higher than 30 dB above the threshold. In the former species there is a statistically significant linear relation between the A₂-cell response and the decrease of the A₁-cell response, but this is not the case inM. latipes (Fig. 3). The similarity of the responses ofE. odorata to those described inEmpyreuma pugione (Coro and Pérez 1984) suggest that also in this noctuid species one may assume that the A₂ cell inhibits the A₁ receptor. In all of these moth species there is a maximum firing rate of the auditory cells at the beginning of the response to pure tones of 5 s and an exponential decrease of their discharge frequency with the course of time (Fig. 5). The analysed species differ in the adaptation rates of their auditory receptors. In all of these species the A₂ cell adapts more rapidly than the A₁ cell. In most of these species the stimulus intensity influences the adaptation rate of the auditory receptors (Fig. 7). These results are compared with data obtained by other authors, and it is concluded that there are more interspecific differences in the physiological characteristics of the auditory receptors in noctuoid species than those reported so far.Abbreviation AP action potential     

Analysis of a cell cycle model based on unequal division of metabolic constituents to daughter cells during cytokinesis

We demonstrate that the unequal division of RNA during cytokinesis explains the dispersion of cell generation times in CHO cell cultures. Experimental cytometric results reported previously serve as a basis for a probabilistic model of cytokinesis. Unequal RNA division to daughter cells, together with two simple laws of RNA production, are used as a source of randomness within the cell cycle. The model reproduces the experimental growth of the CHO cell population, including the observed variability in RNA content. The model has stabilizing properties which explain why a cell population with increased RNA content characteristics, a few cell cycles, to the original pattern. Other cell cycle characteristics, like sister-to-sister and mother-to-daughter generation time correlations implied by the model, are close to their experimental analogs. The conceptual basis of the model is general enough to include unequal division of factors other than RNA (cell mass, cell proteins, etc.) as sources of generation time variability. It seems that the observed dispersion of cell generation times, explained previously in the terms of random transitions in some part of the cell cycle (the Smith & Martin A and B state hypothesis), can be reduced to the single random event of unequal division. This supplies a new convenient tool in the investigation of cell cycle kinetics.     

Acquisition of serum antibodies to specific viral glycoproteins of parainfluenza virus 3 in children.

A radioimmunoprecipitation assay was used to study antibody responses to parainfluenza virus 3 glycoproteins in human sera. The method was not only more sensitive than the neutralization test for the detection of antibody but also provided semiquantitative assessments of the antibody response to both glycoproteins in a single assay system. Anti-hemagglutinin-neuraminidase titers were consistently higher than anti-fusion levels in the same serum specimen. Thirteen children were monitored serologically and virologically from birth until 12 months or more after their primary infection with parainfluenza virus 3. At 1 to 3 months after infection, a significant increase in the level of antibody to the hemagglutinin-neuraminidase protein developed in 12 children; of these, 9 showed rises in the level of fusion protein. In 11 of the children, antibody titers continued to rise and the geometric mean titers to the hemagglutinin-neuraminidase protein was highest in sera collected 8 to 10 months after primary infection. Reinfection as the reason for these progressive increases in antibody levels could only be confirmed for four of the children. Three other children had reinfections after the 10-month sera were obtained; in each instance the only antibody responses were to the fusion protein.     

A quantitative analysis of C3 binding to O-antigen capsule, lipopolysaccharide, and outer membrane protein of E. coli 0111B4

The binding of serum C3 to the O-antigen capsule (OAg Cap), lipopolysaccharide (LPS), and outer membrane proteins (OMP) of Escherichia coli 0111B4 was examined. Bacteria were intrinsically labeled with [3H] or [14C]galactose (*gal) in the OAg Cap and LPS moieties or with [14C]leucine (*leu) to label proteins. Organisms were then incubated in serum containing differentially labeled C3, the above fractions were separated, and the proportion of each binding to a column containing anti-C3 was measured. The OAg Cap fraction bound 72 to 82% of the C3, which bound to E. coli 0111B4 during incubation in absorbed 10% pooled normal human serum (10% PNHS) or absorbed 40% C8-deficient serum (C8D). This distribution did not change when the organism was presensitized with immune IgG before serum incubation. A total of 2.93% +/- 0.48 of OAg Cap and 0.52% +/- 0.16 of LPS *gal bound specifically to Sepharose-containing antibodies to C3 (A:C3-Seph) after incubation in 10% PNHS; these values increased to 10.1% +/- 4.5 and 1.8% +/- 0.3, respectively, when C3 deposition was increased fourfold by incubation in 40% C8D. When encapsulated E. coli 0111B4 was incubated in 10% PNHS containing biotinylated C3, specific attachment of OAg Cap *gal to avidin-Sepharose was demonstrated in 1% sodium dodecyl sulfate (SDS), and complete release of bound *gal but not C3 occurred with 1 M NH2OH. When a mutant of E. coli 0111B4 lacking OAg Cap was incubated in 40% C8D, the outer membrane (OM) bound 85% of C3. Five percent of OM *gal from the unencapsulated organism bound to A:C3-Seph in 0.05% SDS, indicating that the fraction of LPS molecules with bound C3 increased threefold in the absence of OAg Cap. OAg Cap does not contain protein, and no net specific binding of *leu from OAg Cap fractions to A:C3 was detectable; 2.4 to 3.6% of OM *leu bound to A:C3-Seph. Immunoprecipitation of 82.9% of OAg Cap *gal with antisera that were directed to E. coli 0111B4 was associated with co-precipitation of 69.5% of C3 in the capsular fraction. Therefore, the majority of C3 bound to E. coli 0111B4 was covalently attached to OAg Cap and LPS. As corroboration of experiments with whole bacteria, purified OAg Cap and purified LPS consumed C3 when incubated in serum in the fluid phase. These results are the first to evaluate the acceptor site for C3 deposition on a Gram-negative organism incubated in serum, and show that LPS, OAg Cap, and OMP are all major acceptor sites for C3 in nonimmune serum.     

The scrotal myocutaneous flap

The scrotum is a thermoregulatory, well-vascularized structure formed by skin and nonstriated muscle with unique elastic properties. This makes it an ideal source of tissue coverage for problem wounds in its vicinity. Two patients in which scrotal musculocutaneous flaps were used are reported: one, a paraplegic, with a recurrent ischioperineal decubitus ulcer, and another with an ulcer of the penis with exposed Dacron graft previously placed to treat Peyronie's disease. After reviewing the anatomy of the scrotum and the existent literature, we studied scrotal vascularity in a fresh specimen by transillumination. Based on our experience, we conclude that this flap is easy to perform, reliable, and very useful for wounds around the perineal region.     

Hierarchical selection theory and sex ratios I. General solutions for structured populations

Models of sex-ratio evolution in structured populations are derived with G.R. Price's covariance form for the hierarchical analysis of natural selection (1970, Nature 227, 520-521). Previous work on competition among related males for mates (local mate competition), competition among related females for a limiting resource (local resource competition), inbreeding, group selection, and asymmetry of genetic inheritance between males and females, are subsumed under a general formulation for sex-ratio biases in structured populations. I found that the evolutionarily stable strategy sex ratio (males:females) for diploids is 1 - rho m:1 - rho f, where rho m is the regression coefficient of relatedness of the controlling genotypes on males competing for mates, rho f is the regression of controlling genotypes on females that compete for a fixed, limiting resource, and there is no inbreeding. For inbreeding and no competition among females, the evolutionarily stable strategy is 1 - rho m:1 + rho mf, where rho mf is the regression of controlling genotypes on females' mates.