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Biomedical subjects

J A Lindsay

Publications and source records attributed to J A Lindsay.

At least 19 recordsLinked to original sources
PubMed2002

Molecular characterisation of the dominant UK methicillin-resistant Staphylococcus aureus strains, EMRSA-15 and EMRSA-16.

Epidemic methicillin-resistant Staphylococcus aureus types 15 and 16 (EMRSA-15 and EMRSA-16) are the dominant types of MRSA found in UK hospitals, but accurate designation of strains has been difficult. Restriction fragment length polymorphism (RFLP) profiles of seven core virulence genes were used to classify unambiguously isolates of MRSA from St George's Hospital into two groups corresponding to EMRSA-15 and EMRSA-16. Variants of both EMRSA-15 and EMRSA-16 isolates occurred that had lost virulence genes encoded on mobile genetic elements. EMRSA-16 isolates had core gene profiles identical to a cluster of previously characterised MSSA (methicillin-sensitive S. aureus) isolates from St George's Hospital, suggesting that they have arisen from this source, or that loss of the accessory genetic element encoding methicillin resistance is frequent. EMRSA-15 and EMRSA-16 strains were distinct from other MRSA strains previously identified in UK hospitals, and always carried a mobile genetic element encoding multiple superantigens. These results contribute to the understanding of the types of MRSA found in UK hospitals, how they vary and how they arose.

Blotting, Southern↗
PubMed2001

Genetic variation among hospital isolates of methicillin-sensitive Staphylococcus aureus: evidence for horizontal transfer of virulence genes.

Staphylococcus aureus strains often carry in their genomes virulence genes that are not found in all strains and that may be carried on discrete genetic elements. Strains also differ in that they carry one of four classes of an accessory gene regulator (agr) locus, an operon that regulates virulence factor expression and that has been proposed to be a therapeutic target. To look at their distribution among hospital strains, we investigated 38 methicillin-sensitive S. aureus isolates, classifying the isolates by agr class and screening them for the presence and restriction fragment length polymorphisms (RFLPs) of 12 core and 14 accessory virulence genes. Twenty-three (61%) were agr class I, 10 (26%) were agr class II, and 5 (13%) were agr class III. None were agr class IV. The S. aureus strains had distinguishable RFLP profiles, although clusters of isolates with clearly related core gene profiles were found among our strains, including all five agr class III strains, two sets of six strains within agr class I, and six strains within agr class II. Within these clusters there was evidence of horizontal acquisition and/or loss of multiple accessory virulence genes. Furthermore, two isolates from the same patient were identical except for the presence of the sea gene, indicating that movement of mobile elements may occur in vivo. Several strong correlations with the carriage of virulence genes between strains were seen, including a positive correlation between tst and agr class III and negative correlations between tst and lukE-splB and between lukE-splB and seg-sei. This suggests that the core genome or the presence of accessory genetic elements within a strain may influence acquisition and loss of other elements encoding virulence genes.

Bacterial Proteins↗
PubMed2000

Peptic ulcer irony.

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Genes, Bacterial↗
PubMed1999

Evidence for Clostridium perfringens enterotoxin (CPE) inducing a mitogenic and cytokine response in vitro and a cytokine response in vivo.

We investigated some immunogenic properties of Clostridium perfringens enterotoxin (CPE) in vitro using murine J774A macrophages (MPhi) and in vivo using Swiss Webster (SW) mice. CPE was a potent mitogen in vitro, where cell proliferation increased with CPE concentration. CPE was nonmitogenic when MPhi were concurrently incubated with CPE and interferon gamma (IFN-gamma). MPhi incubated in the presence of CPE induced the synthesis of interleukin-1 (IL-1), interleukin-6 (IL-6), tumor necrosis factor alpha (TNF-alpha) and interferon-gamma (IFN-gamma), but not interleukin-2 (IL-2). In vivo, CPE induced a pro-inflammatory cytokine response with striking production of IFN-gamma, IL-1, and IL-6. Regardless of route of CPE entry, serum cytokine levels generally peaked within 1 h of administration and were maintained for 4-8 h. Although CPE engenders an intense immune response during toxicosis, the toxin does not appear to be a superantigen. Death from CPE-induced shock appears to result from various interrelating immunological mechanisms.

Animals↗
PubMed1999

Interactive regulatory pathways control virulence determinant production and stability in response to environmental conditions in Staphylococcus aureus.

The accessory gene regulator (agr) and staphylococcal accessory regulator (sar) loci are important regulators of toxin production in Staphylococcus aureus. In this study we examined how environmental conditions degree of aeration and salt concentration - affect the transcription and translation of mRNAs for alpha-haemolysin (Hla) and serine protease (Ssp) via these pathways and influence the stability of these proteins. Using Northern analysis, we have confirmed earlier observations that sarA is involved in the upregulation of RNAIII, the effector molecule encoded by the agr locus. However, this effect was abolished in highly aerated cultures. While sarA does appear to have an up-regulatory effect on hla transcription that is independent of agr, we propose that the PC1839 (sarA) mutant produces less alpha-haemolysin activity mainly as a result of post-translational inactivation by proteases. The most obvious phenotypic feature of PC1839 (sarA) is the upregulation of proteases. In this study we show that ssp is repressed by SarA at the transcriptional level. Western analysis using an anti-alpha-haemolysin antibody identified a major breakdown product that is only present in the supernatant of strains that are overexpressing serine protease. We have also confirmed that agr exerts a significant regulatory influence on hla at the level of translation, as well as transcription. Finally, the addition of salt upregulates ssp transcription and dramatically downregulates transcription of hla, and is an example of an environmental parameter that affects toxin production independently of agr and sarA. How environmental signals are transduced to control alpha-haemolysin and serine protease production, activity and stability at multiple levels are discussed.

Bacterial Proteins↗
PubMed1998

The gene for toxic shock toxin is carried by a family of mobile pathogenicity islands in Staphylococcus aureus.

Tst, the gene for toxic shock syndrome toxin-1 (TSST-1), is part of a 15.2 kb genetic element in Staphylococcus aureus that is absent in TSST-1-negative strains. The prototype, in RN4282, is flanked by a 17 nucleotide direct repeat and contains genes for a second possible superantigen toxin, a Dichelobacter nodosus VapE homologue and a putative integrase. It is readily transferred to a recA recipient, and it always inserts into a unique chromosomal copy of the 17 nucleotide sequence in the same orientation. It is excised and circularized by staphylococcal phages phi13 and 80alpha and replicates during the growth of the latter, which transduces it at very high frequency. Because of its site and orientation specificity and because it lacks other identifiable phage-like genes, we consider it to be a pathogenicity island (PI) rather than a transposon or a defective phage. The tst element in RN4282, near tyrB, is designated SaPI1. That in RN3984 in the trp region is only partially homologous to SaPI1 and is excised by phage 80 but not by 80alpha. It is designated SaPI2. These PIs are the first in any gram-positive species and the first for which mobility has been demonstrated. Their mobility may be responsible for the spread of TSST-1 production among S. aureus strains.

Amino Acid Sequence↗
PubMed1997

Chronic health effects of microbial foodborne disease.

The acute effects of foodborne disease are sometimes not the end of the illness. Several significant foodborne pathogens are capable of triggering chronic disease, and even permanent tissue or organ destruction, probably via immune mechanisms. Arthritis, septic and reactive, inflammatory bowel disease, haemolytic uraemic syndrome, Guillain-Barré syndrome, and possible several autoimmune disorders can be triggered by foodborne pathogens or their toxins. Research is needed to more fully understand the mechanisms by which the immune system is inappropriately activated by these common foodborne disease-causing agents.

Autoimmune Diseases↗
PubMed1996

Clostridium perfringens type A enterotoxin (CPE): more than just explosive diarrhea.

The bacterial pathogen Clostridium perfringens is the most prolific toxin-producing species within the clostridial group. The toxins are responsible for a wide variety of human and veterinary diseases, many of which are lethal. C. perfringens type A strains are also associated with one of the most common forms of food-borne illness (FBI). The toxicosis results from the production and gastrointestinal absorption of a protein-enterotoxin known as CPE. The regulation, expression, and mechanism of action of CPE has been of considerable interest as the protein is unique. CPE expression is sporulation associated, although the mechanism of cpe-gene regulation is not fully elucidated. Cloning studies suggest the involvement of global regulators, but these have not been identified. Although very few type A strains are naturally enterotoxigenic, the cpe gene appears highly conserved. In FBI strains, cpe is chromosomally encoded; whereas in veterinary strains, cpe may be plasmid-encoded. Variation in cpe location suggests the involvement of transposable genetic element(s). CPE-like proteins are produced by some C. perfringens types C and D; and silent remnants of the cpe gene can be found in C. perfringens type E strains associated with the iota toxin gene. CPE has received attention for its biomedical importance. The toxin has been implicated in sudden infant death syndrome (SIDS) because of its superantigenic nature. CPE can destroy a wide variety of cell types both in vitro and in vivo, suggesting that it could have potential in the construction of immunotoxins to neoplastic cells. It is obvious that CPE is an interesting protein that deserves continued attention.

Amino Acid Sequence↗
PubMed1995

Staphylococcus aureus but not Staphylococcus epidermidis can acquire iron from transferrin.

Staphylococci grow and cause infection under the iron-restricted conditions found in vivo. They therefore must possess mechanisms to obtain iron for metabolism from this environment. To determine if staphylococci can extract iron bound to human transferrin, we labelled transferrin with 55Fe and performed uptake assays on cells grown in iron-restricted and iron-plentiful conditions. Growing cultures of Staphylococcus aureus NCTC 8532 could take up radioactive iron during mid- to late-exponential phase of growth. This process was iron-regulated and did not require direct contact between the cell and the labelled transferrin. Siderophore production was detected during this phase, but reductase or protease activity was not. S. epidermidis ATCC 14990 could not access 55Fe bound to transferrin, nor did this isolate produce siderophore, reductase or protease. This difference in the ability to acquire iron bound to transferrin may contribute to the increased virulence of S. aureus when compared to S. epidermidis.

Biological Transport↗
PubMed1995

Is thermophily a transferrable property in bacteria?

Bacteria exhibit unique diversity in their ability to grow at different temperatures. Indeed, eubacteria and archaebacteria are the only organisms able to grow above 65 degrees C. The temperature range for a species is generally considered to be a stable character; however, mutants may be isolated that have a Tmin or Tmax below or above the parent organism. Some bacteria may also be coaxed to grow at different temperature by training cultures, through an incremental increase or decrease of temperature. Genetic approaches, for example, the transformation of mesophilic Bacillus to thermophily using DNA from closely related thermophiles, has been very controversial. A major problem has been the lack of stability of the high-temperature phenotype upon subculture, which has not allowed extensive genetic and biochemical characterization of the transformants. The mechanism whereby the thermophilic phenotype is carried is unknown, although it is possible that the adapter genes are plasmid encoded. Studies using phenotypically stable transformants indicated that the thermostability of some cellular components was significantly increased, both in the vegetative cell and spore state. Enzyme thermostability, for example, appeared to be associated with an increased use of hydrophobic amino acids; however, the biochemical mechanisms for these alterations remain unknown. Thermophily is still a challenging problem with some interesting molecular biology.

Adaptation, Physiological↗
PubMed1994

Production of siderophore by coagulase-negative staphylococci and its relation to virulence.

The ability to produce siderophore is considered to be a virulence factor for many pathogenic bacteria. To determine if siderophore production by coagulase-negative staphylococci (CNS) was related to virulence, 40 clinical isolates of CNS cultured from peritoneal dialysis fluid were compared with 38 commensal skin isolates. Siderophore activity was detected using the chrome azurol S liquid assay. Using precursor studies, Staphylococcus epidermidis isolates were shown to be more likely to produce the siderophore staphyloferrin A. Production of staphyloferrin B amongst non-Staphylococcus epidermidis species was associated with clinical isolates rather than commensal isolates, and therefore may play a role in pathogenicity.

Citrates↗
PubMed1994

Sudden infant death syndrome (SIDS): are common bacterial toxins responsible, and do they have a vaccine potential?

Despite extensive research, no unifying concept has satisfactorily explained the cause of the sudden infant death syndrome (SIDS). The details are briefly outlined of some of the evidence supporting the hypothesis that common bacterial toxins are important in the aetiology of SIDS. These bacterial toxins act as triggers to initiate a biochemical cascade resulting in death. Data from four research groups, each working independently, indicated that the bacteria Clostridium perfringens, Escherichia coli, Staphylococcus aureus, Streptococcus spp. and Enterococcus spp. were present in higher numbers in infants who had suffered SIDS than in control infants. Certainly more detailed studies need to be performed on the role of bacterial infections in infants. There are many implications arising from this work, particularly the use of vaccination as a means of reducing infections, and consequently the number of SIDS deaths.

Bacterial Toxins↗
PubMed1994

Can superantigens trigger sudden infant death?

A majority of sudden infant death syndrome (SIDS) victims have respiratory or gastrointestinal infections prior to death. This has led to an investigation of the role of pathogenic bacteria and the potentially lethal toxins they produce as triggers for sudden infant death. A small group of bacteria have been consistently identified in SIDS victims as compared to controls, and remarkably, three of these produce superantigenic toxins. Superantigens exert a powerful effect on the immune system, stimulating T-cells, which subsequently induces the formation of large amounts of cytokines. Generation of an overwhelming inflammatory response may lead to death by shock, or other, as yet unrecognized effects of the toxin on the respiratory or cardiac systems. A SIDS/superantigen model is proposed which may explain many of the pathological characteristics of SIDS and establish quantifiable markers for SIDS.

Bacterial Infections↗
PubMed1994

Staphylococcal iron requirements, siderophore production, and iron-regulated protein expression.

Despite the ability of staphylococci to grow in iron-restricted conditions in vivo, their iron requirements and the mechanisms possessed by them for the uptake of iron are poorly understood. Many bacteria are known to produce siderophores. By using the chrome azurol S universal method for the detection of siderophores, all 14 isolates of Staphylococcus aureus tested grew well under conditions of iron restriction and produced iron-regulated siderophore in large quantities, while all 19 isolates of coagulase-negative staphylococci (CoNS) grew poorly under conditions of iron restriction and produced low levels of iron chelator. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis profiles of S. aureus isolates revealed altered protein patterns due to iron restriction, while altered profiles were not seen in the CoNS group. The ability to grow in iron-restricted conditions, possibly with the assistance of siderophore-mediated iron uptake, may contribute to the increased pathogenicity of S. aureus when compared with that of the CoNS.

Bacterial Outer Membrane Proteins↗
PubMed1993

Cloning of the cyclomaltodextrinase gene from Bacillus subtilis high-temperature growth transformant H-17.

The cyclomaltodextrinase gene from Bacillus subtilis high-temperature growth transformant H-17 was cloned on separate PstI, BamHI, and EcoRI fragments into the plasmid vector pUC18, but was expressed in an inactive form in the host, Escherichia coli DH5 alpha. High level constitutive expression of the gene product was also detrimental to the E. coli host, which led to structural instability of the recombinant plasmid. The cyclomaltodextrinase gene was cloned on a 3-kb EcoRI fragment into the plasmid vector pPL708, and the fragment was structurally maintained in the host B. subtilis YB886. The cloned gene product was synthesized in an enzymatically active form in the B. subtilis host; however, expression was at a low level. Subcloning of the 3-kb EcoRI fragment into pUC18 and transformation into E. coli XL1-Blue (F' lacIq) indicated that the cyclomaltodextrinase gene was cloned with its own promoter, since expression of the gene occurred in the absence of IPTG. Subcloning of the cyclomaltodextrinase gene downstream from the Bacillus temperature phage SPO2 promoter of pPL708 may increase expression of this gene.

Amino Acid Sequence↗
PubMed1993

Identification of Staphylococcus epidermidis and Staphylococcus hominis from blood cultures by testing susceptibility to desferrioxamine.

Testing susceptibility to desferrioxamine has recently been described as a method for the identification of Staphylococcus epidermidis. This method was compared to a commercial test and the tube coagulase test for the identification of staphylococci from blood cultures and other fluid specimens. A total of 216 isolates was tested over a 13-month period. Sensitivity of the desferrioxamine test in identifying isolates of Staphylococcus epidermidis and Staphylococcus hominis was 97.3%, while specificity was 91.8%. When isolates displaying discrepant desferrioxamine results were characterized using recently described interpretive criteria, sensitivity and specificity of the desferrioxamine test improved to 100%. The desferrioxamine test was reliable, inexpensive and simple to perform, and should prove useful in the diagnostic laboratory.

Bacteremia↗
PubMed1993

Success and failure rates of cylinder implants in type IV bone.

It is well documented that implant success rates are much lower in Type IV bone than in Types I, II, and III. However, all of these studies have reported on the use of screw type implants. The present study demonstrates the efficacy of cylinder implant use in Type IV bone. A total of 513 cylinders were placed in Type IV bone and were followed for 0 to 60+ months in function. The absolute success rate was 95.7%. Of the 22 fixture failures encountered, 8 were discovered at the time of second stage surgery, 3 during the first year following uncovering, and 11 occurred after 24 to 36 months in function. A possible rationale for the higher success rate of cylinders as compared to that reported for screw type implants in Type IV bone is presented. Further well-controlled studies directly comparing various implant designs in a range of bony sites are needed to better understand and to fully document the differences suggested by the present study.

Alveolar Bone Loss↗