Molecular studies of FIme resistance plasmids, particularly in epidemic Salmonella typhimurium.
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Biomedical subjects
Publications and source records attributed to H R Smith.
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Strains of Salmonella typhimurium of predominantly Middle Eastern origin, but distributed from England to India, were found to carry at least three types of resistance plasmid. The most important was initially identified as an F(I) plasmid by compatibility tests, but differs from the F factor on the one hand and the F(I) factors R162 and ColV on the other. The three groups of F(I) plasmids can be distinguished by their compatibility reactions with the MP10 plasmid of S. typhimurium (Smith, Humphreys, Grindley, Grindley & Anderson, 1973) and group H(1) factors: the F factor is unilaterally incompatible with group H(1) (Smith, Grindley, Humphreys & Anderson, 1973; Anderson, 1975b); the F(I) factors are compatible with MP10 and group H(1); and F(I)me factors are incompatible with MP10 but compatible with H(1). The majority of S. typhimurium cultures belonged to phage type 208; most of those that did not, belonged to types related to 208. Only a minority of their F(I)me plasmids were autotransferring. The remainder were mobilizable by F-like plasmids, and by group H(1) and H(2) factors, but not by the fi(-) I(1) factor Delta, or by plasmids of the I(2), B, P, W, N and com 7 groups. The compatibility reactions of the autotransferring F(I)me plasmids were identical with those of the non-transferring members of the group, and both were large, single-copy plasmids.The S. typhimurium strains of this series carried A or AK, and SSu resistance determinants: small, probably multicopy, non-transferring plasmids similar to those originally described in phage type 29 of S. typhimurium (Anderson & Lewis, 1965b).These S. typhimurium cultures probably represent a clone of wide geographical distribution. The accurate epidemiological study of such clonal outbreaks requires, in addition to phage typing, precise identification of the plasmids harboured by the epidemic strains, and may have to be carried to the molecular level.F(I)me plasmids were identified in other drug-resistant salmonellas, notably in a strain of S. wien which caused large outbreaks of mainly paediatric infection in Algeria, and also spread to Britain. An F(I)me plasmid was found in S. typhi phage type 44 from Algeria, in which the phage-restricting properties of the plasmid are responsible for the specificity of the type.
An investigation of in vitro mutagenesis of plasmid DNA with hydroxylamine is described. The treated plasmid DNA was used to transform Escherichia coli K12. Mutants of the plasmid NTP3, which codes for resistance to ampicillin and sulphonamides, were isolated and characterised. They were classified according to the reduction in level of their beta-lactamase activity. Hydroxylamine-induced mutants of NTP14 were also isolated. This plasmid codes for ampicillin resistance, synthesis of colicin E1, and the EcoRI restriction and modification enzymes. One class of mutants is lethal to the host strain at temperatures above 33 degrees C, but carrier strains grow well at 28 degrees C. There is evidence that these mutants code for a temperature-sensitive EcoRI modification activity: the lethal effect probably results from the cleavage of the host-cell DNA by the restriction enzyme at non-permissive temperatures. The possible genetic uses of the mutant plasmids for the production of hybrid plasmids in the bacterial cell are discussed.
The properties of two plasmids coding for the CcoRI restriction and modification enzymes are described. Both plasmids are non auto-transferring (NTP) but can be mobilised by transfer factors. Strains carrying NTP13 produce colicin E1 and the EcoRI enzymes. This plasmid has a molecular weight of 6 X 10(6) daltons and is present as approximately 12 copies per chromosome. The second plasmid, NTP14, was detected after mobilisation of the EcoRI plasmid with the R factor RI-19. NTP14 codes for ampicillin resistance, synthesis of the EcoRI enzymes and colicin E1. The molecular weight of NTP14 is 10.7 X 10(6) daltons and there are about 14 copies per chromosome. DNA-DNA reassociation experiments were performed to determine the interrelationships of NTP13, NTP14, ColE1 and the R factor R1-19. NTP13 and NTP14 continue to replicate when cellular protein synthesis is inhibited by the addition of chloramphenicol.
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The four R factors described in this paper form a single compatibility group which has been previously designated group H. Recombination was demonstrated between any pair of the plasmids TP117, 123, and 124. In contrast, TP116 did not appear to recombine with any other members of the group. TP117, 123, and 124 usually displaced the F factor from Escherichia coli K-12F(+), while TP116 and F coexisted stably in that strain. Deoxyribonucleic acid reassociation experiments showed minimal homology between F and the four group H plasmids. The results indicate that there are limitations to using incompatibility alone for classification of bacterial plasmids.
Chloramphenicol resistance has been reported in individual strains of Salmonella typhi since 1950, but there had been no accounts of epidemics caused by resistant strains of the organism until 1972, when one occurred in Mexico. Two British patients have been infected in that country, and the organism isolated from them corresponds in all respects with the description of the Mexican epidemic strain of S. typhi: it is a degraded Vi strain resistant to chloramphenicol, streptomycin, sulphonamides, and tetracyclines. It owes its resistance to an R factor which can be transferred to Escherichia coli and thence to drugsensitive S. typhi. Although there is a negligible risk of dissemination of this strain in Britain the incident is a reminder of the need to restrict the use of drugs such as chloramphenicol to the serious diseases for which they are virtually specific.
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The accuracy of skin lesion description by 24 general practitioners was assessed as they used a computer diagnosis assistance system (DERMIS). Descriptive accuracy determines the quality of advice. Only four doctors showed a trend to accurate description over 25 hospital referrals.
Individuals vary in their ability to react to irritants, which can be demonstrated for sodium lauryl sulfate (SLS) using the irritant threshold (IT) test. We aimed to study whether the histological and immunohistochemical features of the skin following SLS exposure varied with subject's IT. 8 subjects were recruited. Their IT was measured. Biopsies were taken after 2 hr and 4 hr of occlusion with 20% SLS and control. The specimens were stained with haematoxylin and eosin and for Langerhans cells. At 4-hr, low-threshold subjects developed changes to a greater extent than high-threshold subjects. The relationship of histological reaction to IT could be related to a differential pro-inflammatory cytokine response in subjects. Low IT has been previously associated with a tumour necrosis factor alpha promoter region polymorphism.
We have developed a simple, efficient procedure for the long term freezing of Tetrahymena thermophila in liquid nitrogen. This technique yields excellent recovery of viable cells with all strains tested and does not require the use of a controlled rate low temperature freezer. To optimize the freezing technique, we have examined the effects of varying a number of parameters, including the physiological state of the cells prior to freezing, the time of exposure to cryoprotectant, and the rate of freezing and thawing. The frequency of viable cell recovery following freezing using this technique has been tested for a variety of different cell lines.