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Biomedical subjects

M Gold

Publications and source records attributed to M Gold.

At least 163 records · Page 9Linked to original sources

Comparison between the prevalence and treatment of wheezing and coughing in Brampton and Mississauga children.

An analysis of hospital discharge data showed that Brampton children 4 years and under had higher rates of hospital admissions for asthma, bronchitis and upper respiratory infections than children in the same age group living in Mississauga and other Ontario municipalities. The present study was done to compare the prevalence and treatment of wheezing and coughing between Brampton and Mississauga children 4 and 5 years of age. The study showed that increased use of pediatricians on the part of Mississauga children in comparison to Brampton children resulted in a higher proportion being placed on anti-asthmatic medications even though the underlying prevalence rate of wheezing and coughing was the same in the two samples. Further study is required to determine whether differences in primary care between the two samples can explain a two-fold increase in hospital admission rates for lower respiratory illness among Brampton preschool children in comparison to their counterparts in Mississauga.

Child, Preschool↗

Anorexia and bulimia.

Anorexia nervosa and bulimia nervosa are eating disorders with distinct clinical presentations. Reduced caloric intake, a hallmark of both disorders, is manifested by self-induced starvation in anorexia and by binge eating and gastrointestinal purging in bulimia. Treatment includes nutritional intervention, psychotherapy and pharmacotherapy in either the ambulatory or the hospital setting.

Adolescent↗

On the effects of replacing the carboxylate-binding arginine-171 by hydrophobic tyrosine or tryptophan residues in the L-lactate dehydrogenase from Bacillus stearothermophilus.

For L-lactate dehydrogenases (LDH's), the interaction of the guanidinium group of their Arg 171 residue with the carboxylate group of an alpha-keto acid is of primary importance in orienting the substrate productively at the active site. LDH's such as that of Bacillus stearothermophilus (BSLDH) are of practical importance for the preparation of chiral 2-hydroxy acids used as synthons in asymmetric synthesis but would even be more valuable in this regard if their specificities were broader. With a view to tailoring the specificity of BSLDH toward carbonyl substrates that lack an alpha-carboxyl group such as ketones, site-directed mutagenesis has been applied to replace Arg 171 by the approximately isosteric, but hydrophobic, amino acids Tyr and Trp. The mutant enzymes exhibit remarkably good catalytic activities toward representative alpha-keto acids RCOCOOH, where R = Me, Et, n-Pr, n-Bu, and CH2OH, although for the mutant enzymes the kcat/KM's are lower by approximately 10(3)-10(4)-fold than those for native BSLDH. Surprisingly, the 171----Tyr/Trp enzymes are significantly more active than 171----Lys (Hart et al., 1987a), for which an interaction of a positively charged side chain with substrate COO- is retained. Preparative-scale 171----Trp catalyzed reduction of pyruvate gave optically pure L-lactate, showing that L stereospecificity of such LDH enzymes was unaffected by the loss of Arg 171. The retention of L stereospecificity is attributed to secondary polar or hydrogen-bonding associations of Arg 109 and Thr246, respectively, with the substrate COO-function that are of sufficient magnitude to maintain "normal" substrate orientation.(ABSTRACT TRUNCATED AT 250 WORDS)

Arginine↗

On the effect on specificity of Thr246----Gly mutation in L-lactate dehydrogenase of Bacillus sterothermophilus.

The function of the amino acid Thr246 in L-lactate dehydrogenase from Bacillus stearothermophilus has been investigated by site-directed replacement with glycine. Kinetic experiments with a number of 2-oxo acids showed strongly reduced activity for the mutated enzyme. However, the mutant enzyme shows a relative preference for the large hydrophobic sidechains of alpha-keto acids and an even higher specific activity than the wild-type lactate dehydrogenase for the polar oxaloacetate substrate. Graphic analyses indicate that the loss of one hydrogen bond, or intrusion of water into the active site, might be responsible for the reduced activity. The kinetic results suggest that the binding modes of bulky hydrophobic or polar substrates compensate to some degree for the partially disrupted active site.

Amino Acid Sequence↗

Integration host factor (IHF) stimulates binding of the gpNu1 subunit of lambda terminase to cos DNA.

The lambda terminase enzyme binds to the cohesive end sites (cos) of multimeric replicating lambda DNA and introduces staggered nicks to regenerate the 12 bp single-stranded cohesive ends of the mature phage genome. In vitro this endonucleolytic cleavage requires spermidine, magnesium ions, ATP and a host factor. One of the E. coli proteins which can fulfill this latter requirement is Integration Host Factor (IHF). IHF and the gpNu1 subunit of terminase can bind simultaneously to their own specific binding sites at cos. DNase I footprinting experiments suggest that IHF may promote gpNu1 binding. Although no specific gpNu1 binding to the left side of cos can be detected, this DNA segment does play a specific role since a cos fragment that does not include the left side or whose left side is replaced by non-cos sequences, is unable to bind gpNu1 unless either spermidine or IHF is present. Binding studies on the right side of cos using individual or combinations of gpNu1 binding sites I, II and III indicate that binding at sites I and II is not optimal unless site III is present.

Attachment Sites, Microbiological↗

Highly atopic children: formation of IgE antibody to food protein, especially peanut.

Highly atopic infants often form IgE antibodies toward multiple food protein in the first 2 years of life. They begin producing IgE antibody to inhalant allergens between the first and second year of life. We hypothesized that highly atopic children would be at significant risk of sensitization to peanut. We defined high atopy as serum IgE greater than or equal to 10 times 1 SD from normal plus multiple positive RASTs. In this study we have characterized the immunologic status of 141 patients by measuring total serum IgE and specific IgE to several allergens, including peanut. These data demonstrated that, independent of clinical history, a positive RAST to peanut was more common in the highly atopic category compared to the low atopy category. Significantly more patients who were highly atopic and had a positive peanut RAST had a positive RAST for egg or milk compared to low atopic patients. More significantly, 33 of the patients had never knowingly received peanut, yet 21 (63.6%) had a positive RAST for peanut, whereas seven (21.2%) had a peanut antibody in the highest RAST category. All these seven patients were considered highly atopic according to the definition above, and three were younger than 2 years of age. These results suggest that highly atopic infants are at special risk for sensitization to peanut, even when they have never received peanut, and that characterization of immunologic sensitization to milk, egg, and peanut will identify the highly atopic infant.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗

Two types of delinquents and their institutional adjustment.

This research tested a typology classifying 326 incarcerated delinquent boys as buoyant or beset, depending on their level of anxiety and depression at intake. Findings reveal that the two types of boys differed in some aspects of life history and personality, that the buoyant type adjusted better to the institution's group-oriented treatment program, and that buoyant and beset boys responded differentially to some components of the treatment. Implications are drawn regarding the usefulness of the typology for guiding differential treatment.

Adaptation, Psychological↗

The Nu1 subunit of bacteriophage lambda terminase.

The maturation and packaging of bacteriophage lambda DNA are catalyzed by the phage terminase enzyme. Terminase is composed of two protein subunits, gpNu1 and gpA. The holoenzyme is multifunctional in vitro; it binds to and cleaves lambda DNA at the cos site (where cos represents cohesive-end site), packages DNA into lambda proheads, and is also a DNA-dependent ATPase. The genes of the two subunits have been cloned separately into powerful expression vectors which allow for very high levels of protein overproduction. The gpNu1 protein has been purified to homogeneity and has a monomeric molecular weight of 21,200, in close agreement with the Mr of 20,444 expected from its amino acid sequence. Both gel filtration and sedimentation velocity centrifugation indicate that the native gpNu1 protein exists as a Mr greater than 500,000 aggregate. The sequence of the first 20 amino acids and the overall composition both match those predicted by the nucleotide sequence of the Nu1 gene. Purified gpNu1 is able to complement gpA-containing extracts in both lambda DNA packaging and cos cleavage assays. The Nu1 gene amino acid sequence predicts DNA binding by the protein, and gpNu1 does show specific binding to lambda DNA by filter binding assays. Also, as predicted from its sequence, gpNu1 exhibits ATPase activity; but in contrast to the holoenzyme, this activity is DNA-independent.

Amino Acids↗

Terminase host factor: a histone-like E. coli protein which can bind to the cos region of bacteriophage lambda DNA.

Terminase Host Factor (THF), an E. coli protein capable of fulfilling the host factor requirement for in vitro bacteriophage lambda terminase activity, displays properties characteristic of the prokaryotic type II DNA-binding or "histone-like" proteins. It is a 22 K basic, heat- and acid-stable protein which binds non-specifically to various DNAs. Conditions can be established, however, where THF binds preferentially to the cohesive end site (cos) of lambda DNA forming several distinct complexes as visualized by band retardation in polyacrylamide gels. DNase I footprinting reveals that THF can protect several regions of the top strand on the right side (+) of cos but does not bind as well to the left side (-). The binding regions are separated either by unprotected or by DNase I- hypersensitive bases. Under the conditions used in these experiments, DNA which does not contain cos lambda sequences does not show this pattern of protection. Several repeated motifs in the cos lambda nucleotide sequence may represent a consensus sequence for THF interaction. THF may be similar to other "histone-like" proteins which display both non-specific and selective DNA-binding capacities.

Bacteriophage lambda↗

Prediction of an ATP reactive center in the small subunit, gpNu1, of the phage lambda terminase enzyme.

The small subunit of the bacteriophage lambda terminase enzyme, the product of the phage's Nu1 gene, is shown to contain amino acid segments homologous to those present in a large number of ATPases. In keeping with these predictions, the purified protein has been found to hydrolyze ATP with a relatively low turnover number. Terminase holoenzyme is a known ATPase, and the biochemical significance of an ATP-interactive center situated in the gpNu1 subunit is discussed.

Adenosine Triphosphatases↗

Late stages in bacteriophage lambda head morphogenesis: in vitro studies on the action of the bacteriophage lambda D-gene and W-gene products.

The in vitro maturation of bacteriophage lambda can be divided into discrete steps. Concatemers of lambda DNA bind terminase to form complex I. This DNA-terminase complex then binds a prohead to form a ternary complex (II). Complex II in turn can be converted to infectious phage by the addition of extracts containing the products of the phage genes D, W, FII, as well as phage tails. By using in vitro complementation assays gpD and gpW have been partially purified and their interactions with complex II studied. gpD can bind to complex II in vitro to form a new complex (III) which can be isolated by sedimentation on neutral sucrose gradients. This complex requires only the addition of gpW, gpFII, and phage tails to form mature phage particles. The sedimentation of complex III is virtually identical to that of complex II; however, the resistance of the former to inactivation by DNase is higher, likely due to the partial packaging of the DNA. In similar experiments it was shown that gpW cannot bind to complex II but can effectively interact with complex III. This latter reaction converts complex III to a DNase-resistant form which sediments in a manner identical to that of full phage heads (complex IV). After isolation of the complex IV only gpFII and tails are required for mature phage formation in vitro. gpW is a heat-stable protein of molecular weight approximately 10,000.

Bacteriophage lambda↗

The Nul subunit of bacteriophage lambda terminase binds to specific sites in cos DNA.

The maturation and packaging of bacteriophage lambda DNA are under the control of the multifunctional viral terminase enzyme, which is composed of the protein products of Nu1 and A, the two most leftward genes of the phage chromosome. Terminase binds selectively to the cohesive end site (cos) of multimeric replicating lambda DNA and introduces staggered nicks to regenerate the 12-base single-stranded cohesive ends of the mature phage genome. The purified gpNu1 subunit of terminase forms specific complexes with cos lambda DNA. DNase I footprinting experiments showed that gpNu1 bound to three distinct regions near the extreme left end of the lambda chromosome. These regions coincided with two 16-base-pair sequences (CTGTCGTTTCCTTTCT) that were in inverted orientation, as well as a truncated version of this sequence. Bear et al. (J. Virol. 52:966-972,1984) isolated a mutant phage which contained a CG to TA transition at the 10th position of the rightmost 16-base-pair sequence, and this phage (termed lambda cos 154) exhibits a defect in DNA maturation when it replicates in Escherichia coli which is deficient in integration host factor. Footprinting experiments with cos 154 DNA showed that gpNu1 could not bind to the site which contained the mutation but could protect the other two sites. Since the DNA-packaging specificity of terminase resides in the gpNu1 subunit, these studies suggest that terminase uses these three sites as recognition sequences for specific binding to cos lambda.

Bacteriophage lambda↗

Common sense on extending DRG concepts to pay for ambulatory care.

The development of Medicare's prospective payment system has led to considerable interest in expanding case-based payment to other forms of care, including ambulatory care. In applying approaches developed in one form of care to another, one must consider whether the same characteristics and conditions apply. This paper reviews the similarities and differences between ambulatory and inpatient care in light of available research. It identifies critical issues that should be considered in designing ambulatory payment reform and draws conclusions on the appropriateness of extending case-based payment principles to ambulatory care.

Ambulatory Care↗