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Biomedical subjects

S Hayes

Publications and source records attributed to S Hayes.

At least 19 recordsLinked to original sources

Working hours: an employer's duty.

Recent EC negotiations on the Working Time Directive have shown UK thinking to be at odds with the rest of Europe. Susan Hayes examines the implications for employee health.

Employment

Mapping ethanol-induced deletions.

Chromosomal rearrangements, uniformly represented by very large deletions, were stimulated upon transiently exposing Escherichia coli cells with a defective lambda prophage to about 18% (v/v) ethanol. It was shown that the ethanol treatment induced deletion formation rather than enriching for ethanol-tolerant cells. The deletions in 435 mutants were mapped to 26 groups. Ethanol treatment changed the spectrum of deletions relative to those arising spontaneously, and stimulated the formation of deletions with endpoints in E. coli DNA flanking the lambda fragment. The promotion of deletion formation by ethanol involves the joining of distant, nonhomologous linear DNA segments, which can be considered an illegitimate recombination event; however, activity of the E. coli recA gene product was also required. Although spontaneous deletions arose in comparable cells defective for recA, the incidence of deletion formation in recA cells was not altered by ethanol. It is proposed that ethanol stimulates chromosomal rearrangements involving two oppositely oriented replication forks, since the localized deletions commonly removed or inactivated the chromosomal segment including the bidirectional lambda origin of replication. The results imply a novel mutagenic process induced by an agent that does not act directly on DNA.

Bacteriophage lambda

Alcohol treatment of defective lambda lysogens is deletionogenic.

We ascertained that transient exposure to ethanol, above 18%, was deletionogenic to an Escherichia coli strain with a fragment (12.5 kb) of bacteriophage lambda integrated within the chromosome. The lambda attL B.P' through P fragment provided a forward selection for mutants, and a target for mutagenesis. The cells were killed by thermal derepression of transcription and replication of the lambda fragment when transferred from 30 degrees to 42 degrees C. Survivor mutants, capable of forming colonies at 42 degrees C, were selected from untreated starting cells. About half no longer supported marker rescue of the lambda fragment imm lambda (immunity) region, comprising the cI repressor, and the PL and PR promoters. Ethanol treatment of starting cells increased the occurrence of imm lambda-defective clones to near 100%. The mutations responsible for the imm lambda defect were found to be large deletions (12 kb or more of DNA). Ethanol treatment of the starting cells also produced a 5- to 18-fold increase in the occurrence of E. coli pgl mutations, which likely arose by the deletion mechanism generating the imm lambda defects, since pgl was closely linked to the integrated lambda fragment. A unifying hypothesis for these observations was that ethanol was deletionogenic. The inclusion or substitution of the int-kil segment of the lambda fragment produced no real change in the spontaneous occurrence of large imm lambda deletions from the untreated cells. Substitution of this segment suppressed the deletionogenic effect of ethanol, implying a prerequisite for sequence homology or gene function from this interval.(ABSTRACT TRUNCATED AT 250 WORDS)

Bacteriophage lambda

Stimulation of mutations suppressing the loss of replication control by small alcohols.

Transient exposure of lysogenic Escherichia coli cells to small alcohols stimulated the frequency of mutations suppressing the lethal loss of replication control from a prophage fragment of bacteriophage lambda. The stimulation in mutation frequency paralleled the effect of mutagenic agents, and in this sense the alcohols behaved as mutagens. 10-min treatments above distinct threshold concentrations at 23%, 18%, 10% and 4% (v/v) were required in order for methanol, ethanol, isopropanol and propanol to evoke mutagenic effects. The selected mutant cells were, in general, equally or more sensitive to ethanol than the starting cells. The mutagenicity of methanol and ethanol was detected only with E. coli strains with lambda fragments that included the site-specific and general recombination genes found within the phage int-kil gene interval; whereas, stimulation of the frequency of phenotypically identical mutations by nitrosoguanidine or ionizing radiation did not require that the lambda fragment encode these genes. Treatments of lysogenic cells with mutagenic concentrations of ethanol did not trigger prophage induction and were concluded not to induce a cellular SOS response nor to denature the prophage repressor, or to disrupt repressor-operator binding. The toxicity of ethanol was pH-dependent. Cellular sensitivity to ethanol toxicity was unaffected by the integrated lambda fragment(s) or by an intact lambda prophage; but, it was increased by deletions of the E. coli chromosome extending rightward from bio into uvrB, and rightward from chlA.

1-Propanol

What are the 'best' measurements for monitoring patients during short-term second-line therapy?

Seventy-one patients with active rheumatoid arthritis were treated with one of five second-line agents and monitored for 24 weeks using seven clinical and seven laboratory measurements. Statistical tests were used to determine which measures changed fastest, which changed most and which most closely reflected changes in the others. The results consistently showed that articular index and summated change score were the 'best' clinical measures while ESR and plasma viscosity were the 'best' laboratory measures. Traditional measures such as grip strength and joint size fared badly and cannot be recommended. Clinical variables improved slightly more rapidly than laboratory measures, but the latter showed the greater change.

Anti-Inflammatory Agents, Non-Steroidal

The generic shoe: a prescription tool.

A multiple-use testing shoe capable of various orthotic modifications has been developed. Its use in predicting prescription performance and patient compliance is presented. Patient compliance is enhanced by understanding gained through the testing procedure.

Biomechanical Phenomena

pK1' and bicarbonate concentration in plasma.

Values for pK1' were determined from pH measured at 37 degrees C with three blood-gas analyzers and from calculated pco2 values in 443 freshly separated plasmas, tonometered at 37 degrees C. Plasma was taken from healthy volunteers, seriously ill patients, and hyponatremic patients. pK1' values varied by considerably more than 0.06 in healthy volunteers as well as in very ill patients, and bicarbonate concentrations ([HCO3]p) calculated by blood-gas analyzers based on the pK1' value of 6.1 could be in error by some +/- 60%. pK1' was similarly determined for tonometered (37 degrees C) replicate dilutions of plasma samples. By adding weighed amounts of dry NaCl and NaHCO3 to the diluted samples we increased the Na+ concentration to approximately 150 mmol/L and bicarbonate concentrations to values ranging from approximately 2.5 to approximately 52.5 mmol/L. pK1' values decreased when [HCO3]p was increased in dilutions of plasma kept at constant ionic strength. At any given [HCO3]p, pK1' values were higher at high than at low values of pco2.

Bicarbonates

Spontaneous lambda OR mutations suppress inhibition of bacteriophage growth by nonimmune exclusion phenotype of defective lambda prophage.

Survivor clones with defects in gene functions that participate in the replicative killing of thermally induced Escherichia coli constructs with integrated lambda N through P or cIII through P gene fragments were selected at a frequency of about 10(-6). Among the population of survivors, clones were identified that exhibited normal lambda immunity at 30 degrees C, as shown by their ability to prevent the plating of lambda wild type and to support the plating of a nearly identical heteroimmune bacteriophage lambda imm434. However, when placed at 42 degrees C to inactivate the cIts857 repressor, these survivor isolates excluded the plating of both lambda wild-type and lambda imm434 phages, a phenotype designated nonimmune exclusion (Nie). Spontaneous mutants of lambda wild type were isolated that overcame the Nie phenotype and would plaque at 42 degrees C on cell lawns of these isolates. The acquired lambda se mutations suppressed nonimmune exclusion, prevented lysogenization by interrupting repressor expression from PRM, and made the phage insensitive to replicative inhibition. The se mutations were genetically mapped and sequenced within the rightward lambda operator site.

Bacteriophage lambda

Nephrotic syndrome with renal vein thrombosis: pathogenetic importance of a plasmin inhibitor (alpha 2-antiplasmin).

Tests of fibrinolysis were measured by fibrin plate methods in 44 patients with nephrotic syndrome, in 14 of whom renal vein thrombosis was demonstrated. In both groups the level of total fibrinolytic activity was normal, that of vascular plasminogen activator was decreased, and that of an inhibitor of plasminogen activation was elevated. The level of a plasmin inhibitor, measured by the fibrin plate method, was elevated in 13 of 14 patients with, but only in 12 of 30 without, renal vein thrombosis (p less than 0.005). The plasmin inhibitor was identical with alpha 2-antiplasmin. The data suggest that an increased level of alpha 2-antiplasmin may be a factor in determining susceptibility to the development and persistence of renal vein thrombosis in patients with nephrotic syndrome.

Fibrinolysis

Control of bacteriophage lambda repressor establishment transcription: kinetics of l-strand transcription from the y-cII-oop-O-P region.

The kinetics of lambda l-strand repressor establishment RNA synthesis were measured from the y-cII region of induced tof- prophage. The activity of the repressor is epistatic to the expression of gene tof coding for the antirepressor (Tof). The activity of Tof, is epistatic to the expression of repressor gene cI transcription from Prm and the expression of repressor establishment transcription from a site 600 to 800 nucleotides upstream from Prm. Three modes of l-strand rex-cI-tof-y-cII-oop transcription occur: (a) Prm promoted cI-rex mRNA synthesis from noninduced prophage, (b) coordinate lit and oop synthesis from induced tof+ prophage and (c) establishment transcription from induced tof- prophage. The synthesis or stability of oop RNA is much reduced from induced tof-, compared with tof+ prophage. The oop transcription from tof- prophage is not coordinate with RNA synthesis from the y-cII interval. The y-cII-(oop) portion of the establishment transcript appears more unstable than the translated downstream copy of genes rex-cI. The initiation of any repressor establishment transcription requires the products of lambda genes cIII, cII, P and Escherichia coli genes dnaB, dnaG, but not actual lambda DNA synthesis. This result demonstrates that common factors, i.e. replication gene products, are required for the initiation of establishment transcription, lambda replication and lit, oop RNA synthesis; and explains why cIII+ cI+ cII+ replication defective phage lysogenize poorly at low multiplicities of infection. The cIII and cII products were shown to act after an earlier replication initiation or activation event. Repressor establishment transcription and repressor mRNA synthesis from Prm (from induced cI- tof-, cIII- cI- tof- or cI- tof- cii- prophage) are amplified by gene dosage. The extent of lysogenization of E. coli by lambda cIII-, cII- or replication minus mutants, defective for initiation of establishment synthesis, is attributed to gene dosage dependent transcription from Prm. The mechanism by which Tof inhibits the initiation of establishment transcription does not appear to require repression of RNA synthesis from PL and PR. RNA synthesis from these promoters is blocked by renaturation of the repressor 5 min after induction, before establishment transcription is detected; however, establishment RNA synthesis measured between 12-13 min after induction, i.e. 7 min after renaturation of the repressor, is only partially reduced.

Coliphages

Use of ganglioside affinity filters to identify toxigenic strains of Clostridium botulinum types C and D.

Clostridium botulinum neurotoxin is synthesized by toxic clones grown anaerobically on ganglioside affinity filters. The toxin binds to the filters and is detected by reaction with 125I-immunoglobulin G from type-specific antitoxin. Toxin spots from culture filtrates were similarly identified. The C. botulinum type C and D strains were selected for developing this affinity filter assay because synthesis of the C1 and D toxins is bacteriophage dependent. Toxigenic clones were distinguished from prophage-cured atoxigenic derivatives. These studies represent a first step toward the development of a general nonbiological screening procedure for identifying botulinal toxin and toxigenic cells. The affinity filter methodology should facilitate genetic analysis of the basis of C. botulinum toxicity.

Bacteriological Techniques

Ultrasound in the evaluation and diagnosis of jaundice.

Our experience in the evaluation and diagnosis of jaundice by ultrasound in a consecutive series of patients examined in the past year is reported. A final diagnosis is available in 49 patients with obstructive jaundice and 41 patients with nonobstructive jaundice. The accuracy of separation into these two groups is 97% in this series. We would like to stress the value of ultrasound as a complete investigation in many jaundiced patients. In this series a full diagnosis of the cause of jaundice was achieved in 58% of patients.

Adult

Control of lambda repressor prophage and establishment transcription by the product of gene tof.

Control of expression of the bacteriophage lambda (lambda) repressor was studied by measuring repressor transcription in noninduced and derepressed lambda lysogens. Three distinct modes of leftward transcription were observed from cI and the adjacent genes associated with the control of repressor synthesis: The prophage or maintenance mode Prm-cI-rex-ti repressor transcript occurs from repressed lysogens; the oop (Po-oop-to) transcript, and the lit (lit-ti) RNA, from the distal half of gene rex, both occur from induced tof+ prophage; the repressor establishment mode of transcription is observed throughout the rex-cI-tof-y-cII-oop interval between Po and ti from induced tof- prophage. The overall level of establishment mRNA synthesis is partially template dependent. However, the actual initiation step for repressor establishment transcription requires the participation of the lambda cIII, cII products, and also either requires the activity of Escherichia coli replication proteins, or is triggered by a replication initiation event. The cII cIII products do not positively stimulate de novo initiation of establishment transcription, but rather act after an initial replication-dependent step. Initiation of the establishment mode of repressor transcription is totally inhibited by more than 125-fold, in an all or none fashion, by the lambda antirepressor (Tof), the product of gene tof (cro). Since Tof only reduces the in vivo rightward transcription of cII from Pr by about 2-fold, we suggest that Tof inhibits repressor establishment transcription by either uncoupling the replication and cII-cIII dependent events, or by inhibiting the activity rather than the expression of the cIII, cII products. Our results do not fully support either of the present hypotheses that establishment transcription is initiated from the hypothetical Pre promoter in the y-interval, or arises through antitermination of the oop RNA. Since the initiation and control of the establishment mode of repressor transcription parallels the control of lit RNA synthesis, we propose a common mechanism underlies the initiation of these transcripts.

Coliphages

Verbal communication between students in multidisciplinary health teams.

The study reported here was connected with the influence of two variables--the context in which communication occurs, and the concept availability of various health professionals--upon the verbal interaction of various health professionals. Specifically, the verbal output of health professionals in groups consisting entirely of their own professional peers (i.e. all nurses, or all medical students) was compared with that of professionals in multidisciplinary health teams. The study, in attempting to delineate possible barriers to effective verbal interaction, has important implications for the functioning of multidisciplinary health teams.

Australia