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M H Richmond

Publications and source records attributed to M H Richmond.

At least 19 recordsLinked to original sources

'Cells' and 'organisms' as a habitat for DNA.

Although the bulk of the hereditary information in bacteria is organized as a single chromosome, it has been known for some years now that bacteria may also carry pieces of self-replicating extrachromosomal DNA. These units are known as plasmids. Sometimes such plasmids carry the information necessary to give rise to mature bacterial viruses under appropriate conditions, but in other cases they specify the production of enzymes and other proteins which alter the bacterial phenotype. Plasmids are often inessential for survival of bacteria, although they may widen the range of environmental conditions under which they flourish. Thus plasmids may be thought of as adventitious additions to the genetic content of bacterial cells. Recently it has become clear that furthur organizational units of DNA are to be found in bacterial cells. These units are called insertion sequences and transposons. Unlike plasmids and the chromosome, however, these DNA units do not carry enough genetic information to specify their own independent replication: they must rely on plasmids or the chromosome for that purpose. Nevertheless they behave in many respects as independent functional units. Although it is possible to think of the chromosome, plasmids and transposons/insertion sequences as three distinct hierarchies of bacterial DNA, genes may move from one hierarchy to another; and such transitions have important implications for the evolution of bacterial populations. Moreover, their study in bacteria may throw much light on the type of DNA interactions occurring in higher cells.

Base Sequence

Persistence of plasmid-carrying tetracycline-resistant Escherichia coli in a married couple, one of whom was receiving antibiotics.

This paper describes an extension to an earlier account of the coliform flora carried by a married couple, one of whom was taking tetracycline for prolonged periods. The latter phase of this study was notable for the following: first, certain tetracycline-resistant Escherichia coli O antigen types persisted in one of the participants for several weeks after tetracycline was withdrawn; second, a course of ampicillin led to replacement of the tetracycline-resistant flora by one that was ampicillin resistant, but the end of the ampicillin course led to the reappearance of the tetracycline-resistant line, even though no tetracycline was being taken; and third, the tetracycline-sensitive O75 E. coli, which appeared toward the end of the survey, had not lost their plasmid but carried a derivative in which the tetracycline resistance gene(s) had been inactivated by the insertion of an extra piece of deoxyribonucleic acid with a molecular weight of about 1 megadalton.

Antigens, Bacterial

Adhesion of commensal bacteria to the large intestine wall in humans.

Biopsies taken during colonoscopic examination of the human large bowel were used to examine the relationship of the commensal bacterial to the mucosal epithelial cell surface. Bacteria were seen adhering to the exposed epithelial cell surface and also to the mucus sheet. Isolation of aerobic organisms showed that Escherichia coli are closely associated with the gut wall throughout the large intestine. One strain of E. coli predominated in each biopsy, and this strain was present along the whole length of bowel. Adhesion of bacteria to the gut wall does occur in vivo and may be one of the factors involved in the ability of an organism to colonize and persist.

Escherichia coli

Adhesion to a human cell line by Escherichia coli strains isolated during urinary tract infections.

It has been shown that some, but not all, Escherichia coli strains isolated from urine adhere, in vitro, to the surface of uroepithelial or vaginal cells. In the present study, 212 strains, isolated from urine of 212 infected patients, were tested for adhesion by using an in vitro human cell line assay. A variable degree of attachment to the cell monolayer was detected in these strains. From patients with cystitis, only 19 (9.7%) of the 195 strains examined were adherent, whereas 5 (29.4%) of the 17 pyelonephritis strains had similar properties (P less than 0.05). To investigate the incidence of adhesion in the clinical manifestations of urinary tract infection, a sample of patients was picked at random from those with cystitis. During cystitis caused by adhesive bacteria, patients suffer more often from macroscopic hematuria than from dysuria, frequency, or recurrency (P less than 0.05). This study shows that E. coli strains isolated from urine samples possess a strikingly difference in capacity to adhere to a human cell line surface as demonstrated previously with uroepithelial or vaginal cells. Moreover, according to these data, the adhesion of E. coli may be considered as a virulent factor and would play a part in the infection of the urinary tract in humans.

Adhesiveness

Sites of insertion of TnA and TnM in RP1 and its derivatives.

The sites of insertion of TnM into the plasmid RP1, into derivatives of RP1 lacking TnA, and into derivatives in which the location of TnA within RP1 has been altered, were determined. Similarly, the sites of insertion of TnA into derivatives of RP1 from which TnA had been deleted, both with and without copies of TnM, have been examined. These studies show the presence in the plasmids of 'hot-spots' for TnA and TnM insertion. It is clear from the observations, however, that a particular DNA sequence in the recipient replicon is not sufficient to definite a 'hot-spot' since particular sequences sometimes do, and sometimes do not, contain many sites of insertion for a given transposon.

Base Sequence

Beta-lactamases.

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Bacteria

The persistence of R-plasmid-carrying E. coli in a married couple, one of whom was receiving antibiotics.

The aerobic Gram-negative intestinal flora of two individuals, husband and wife, has been followed for about 20 months. The wife was receiving prolonged tetracycline treatment for acne during the first year of the study and was found to carry a large proportion of tetracycline resistant E. coli in her faecal flora even after the tetracycline treatment had ended. A brief therapeutic course of ampicillin during the period when no tetracycline was being taken resulted in the temporary disappearance of the tetracycline resistant flora, but this returned, even though no tetracycline was being taken, as soon as the ampicillin ended. The husband took no antibiotics during the period under study but was frequently found to excrete the same resistant E. coli as his wife. Moreover, the plasmids carried by the tetracycline resistant strains in the wife and the husband were often indistinguishable. This suggests that R-plasmids may spread from people under treatment to close relatives who have not been treated.

Ampicillin

Regional preference of insertion of Tn501 and Tn802 into RP1 and its derivatives.

The sites of insertion of Tn501 into RP1 and into derivatives of this plasmid that either lack the Tn801 (TnA) element or contain it in a different location have been determined. Similarly, the sites of insertion of Tn802 into a derivative of RP1 that lacks the Tn801 element and into recombinants of this plasmid with Tn501 were determined. 'Hot spots' for insertion were observed with both transposons; but it is clear that a particular DNA sequence is not sufficient to define a 'hot spot', since a particular region does contain many insertions when present in one plasmid but does not do so when part of another.

Base Sequence

The stable carriage of two TnA units on a single replicon.

Bacterial plasmids which contain a copy on TnA are refactory to the uptake of a second by transposition. However plasmids containing two such copies can be constructed by in vitro recombination techniques. Some plasmids containing two copies of TnA have been obtained by conventional transposition, but in all cases they arose by the virtually simultaneous insertion of both units into a replicon that carried no TnA. All stable plamids containing two copies of TnA carried the transposons in opposite orientation.

DNA Replication

Fermentation of raffinose by lactose-fermenting strains of Yersinia enterocolitica and by sucrose-fermenting strains of Escherichia coli.

Introduction of plasmids carrying the lacY gene (lactose permease gene) into Yersinia enterocolitica results in cells being able to ferment both lactose and raffinose. Transfer of such plasmids into Escherichia coli C600 (lacY) confers ability to ferment lactose but not raffinose. Derivatives of C600 that ferment both lactose and sucrose (Lac+ Scr+ strains) are able to ferment raffinose, but do not grow well on raffinose minimal medium. Fermentation of raffinose by Lac+ strains of Y. enterocolitica, and by Lac+ Scr+ strains of E. coli, is explained in terms of transport of raffinose via the lac permease and subsequent breakdown catalyzed by invertase.

Conjugation, Genetic

beta-lactamases and R-plasmids of Haemophilus influenzae.

The emergence of resistance to ampicillin and other antibiotics in Haemophilus influenzae has been a relatively recent event. In contrast, drug resistance has been rampant in the Enterobacteriaceae for many years. Ampicillin-resistance in H. influenzae is almost invariably attributable to possession of the TEM (Type III a)beta-lactamase. As is common in other bacteria the gene specifying this enzyme is plasmid-borne in Haemophilus. Some ampicillin-resistant strains of H. influenzae can transfer the TEM beta-lactamase gene to other strains of Haemophilus, to Escherichia coli and to Pseudomonas aeruginosa. The features of such transfer are unusual and lead for example, to the induction of adenine requirement in recipient strains of P. aeruginosa. Crypticity measurements of beta-lactamase activity show that in comparison to P. aeruginosa or E. coli, the outer membrane of H. influenzae affords only a weak penetration barrier to beta-lactam antibiotics. This may have consequences for the stability and distribution of beta-lactamase production in Haemophilus spp. which are discussed. A comparison of the molecular properties of R-plasmids determining a variety of resistances and carried by strains of H. influenzae isolated in diverse geographical locations has revealed unexpected homologies. A series of such plasmids of similar molecular weights (about 30 X 10(6)) differ substantially only in the transposable resistance genes that they carry. A model based on these findings is presented to explain the acquisition of ampicillin- and other resistances by Haemophilus.

Amidohydrolases

beta-lactamase insensitive or inhibitory beta-lactams: two approaches to the challenge of ampicillin-resistant E. coli.

The rising incidence of Gram-negative bacteria resistant to ampicillin in hospitals has brought about a need to develop novel beta-lactam antibiotics. This challenge is being met in two ways. First, attempts have been made to develop compounds insensitive to beta-lactamase hydrolysis. This has proved difficult in the penicillin series, but cefuroxime and cefoxitin are cephalosporin derivatives with the necessary properties. The second approach has been to discover novel beta-lactam nuclei capable of inhibiting beta-lactamases. So far this had led to the production of the clavulins and the olivacins, the latter class of molecules being closely related chemically to thienamycin.

Amidohydrolases