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Biomedical subjects

B Inglis

Publications and source records attributed to B Inglis.

At least 19 recordsLinked to original sources

Expression of rabbit IL-4 by recombinant myxoma viruses enhances virulence and overcomes genetic resistance to myxomatosis.

Rabbit IL-4 was expressed in the virulent standard laboratory strain (SLS) and the attenuated Uriarra (Ur) strain of myxoma virus with the aim of creating a Th2 cytokine environment and inhibiting the development of an antiviral cell-mediated response to myxomatosis in infected rabbits. This allowed testing of a model for genetic resistance to myxomatosis in wild rabbits that have undergone 50 years of natural selection for resistance to myxomatosis. Expression of IL-4 significantly enhanced virulence of both virulent and attenuated virus strains in susceptible (laboratory) and resistant (wild) rabbits. SLS-IL-4 completely overcame genetic resistance in wild rabbits. The pathogenesis of SLS-IL-4 was compared in susceptible and resistant rabbits. The results support a model for resistance to myxomatosis of an enhanced innate immune response controlling virus replication and allowing an effective antiviral cell-mediated immune response to develop in resistant rabbits. Expression of IL-4 did not overcome immunity to myxomatosis induced by immunization.

Animals↗

In vivo 1H magnetic resonance imaging and spectroscopy of the rat spinal cord using an inductively-coupled chronically implanted RF coil.

An inductively coupled, chronically implanted short-solenoid coil was used to obtain in vivo localized 1H NMR spectra and diffusion-weighted images from a rat spinal cord. A 5 x 8 mm two-turn elliptically shaped solenoid coil was implanted in rats at the site of a T-12 vertebral-level laminectomy. Excitation was achieved solely by a 3 x 3 cm external surface coil, and signal detection was achieved by inductively coupling the external coil to the implanted coil. The image signal-to-noise ratio (SNR) obtained with the inductively-coupled implanted coil was compared with that obtained using a linear or a quadrature external surface coil. The implanted coil provided a gain by over a factor of 3 in SNR. The implanted coil was used to measure localized 1H spectra in vivo at the T13/L1 spinal-cord level within a 1.85 x 1.85 x 4.82 mm (16.5 microL) volume. With 256 averages, a approximately 3-s repetition delay and respiratory gating, a high-quality spectrum was acquired in 13 min. In addition, water translational diffusion was measured in three orthogonal directions using a stimulated-echo imaging sequence, with a short echo time (TE), to produce a quantitative map of diffusion in a rat spinal cord in vivo.

Animals↗

Student reflective groups at a Scottish College of Nursing.

In this paper student views on reflective groups, set up as an important element of the new Project 2000 course in a Scottish College of Nursing, are reported. A random sample of 19 students were interviewed. While the reflective groups were very popular with students because they provided support, there was little evidence of a linkage between theory and practice. It was clear that the ambitious objective of stimulating reflection-on-action was not attained. Practice certainly was discussed, but it tended to be dominated by dramatic and emotionally charged aspects of care rather than the more frequent routine concerns. There were, however, indications that the original aim of the reflective groups could be achieved if tutors could establish a common understanding of the purpose of the groups and of reflection, and if the practices on which students reflected consisted less of single day visits where the students saw themselves as nonparticipant outsiders.

Education, Nursing, Baccalaureate↗

Situating theory in practice: student views of theory-practice in Project 2000 nursing programmes.

In this paper the views of student nurses taking Project 2000 at a Scottish College of Nursing are reported. A random sample of 19 students were interviewed. The interviews focused on the relationship between the college course and the students' experiences on placement. The majority had difficulty in relating the college course to their practice, except when it came to very specific parts of the programme, for example basic medical procedures. Only a minority could integrate the broader aspects of the course to their practice, largely because there was such a long period of time between studying a topic in college and the related placement. It is tempting to see the results as supporting the situated cognition hypothesis that theory loses meaning out of context, and it is evident, even from this limited study, that the success of Project 2000, with its emphasis on the 'knowledgeable doer', could depend, to a great extent, on the careful phasing of college coursework and placement to enable students to integrate theory and practice more effectively.

Attitude of Health Personnel↗

IS257 and small plasmid insertions in the mec region of the chromosome of Staphylococcus aureus.

Four copies of the insertion sequence IS257 are found in the mec region of the chromosome of the Australian methicillin resistant Staphylococcus aureus (MRSA) strain ANS46, two flanking a merAmerB sequence (encoding resistance to mercurial compounds), the other two flanking an integrated copy of the plasmid pT181 (tetracycline resistance). The termini of the integrated copy of the plasmid pT181 carry a direct repeat of 8 bp of plasmid sequence, but otherwise there are no similarities in the 8 bp sequences flanking the four copies of IS257 in this strain. Integrated copies of pT181 in strains R35 (a New Jersey MRSA) and GH32 (MRSA of Greek origin) have the same terminal repeat as in ANS46, suggesting either a specific site of insertion of IS257 into the free plasmid before integration into the chromosome, or a common evolutionary lineage for these geographically diverse isolates. A different 8 bp terminal repeat of plasmid sequence is found in the chromosomally integrated copy of pUB110 (flanked by a pair of IS257s) in R155, another New Jersey MRSA. This 8 bp repeat differs from that reported previously for pUB110/IS257 inserted into the plasmid pSK41, indicating insertion of IS257 into different sites of pUB110 before integration into the chromosome or into pSK41. In the plasmid pSK1, the two outer copies of IS257 of the three associated with Tn4003 (trimethoprim resistance) are also flanked by 8 bp repeats.(ABSTRACT TRUNCATED AT 250 WORDS)

Base Sequence↗

Methicillin-sensitive and -resistant homologues of Staphylococcus aureus occur together among clinical isolates.

Clinical isolates of Staphylococcus aureus collected from hospitals in Australia were analyzed for genetic similarities using restriction fragment length polymorphisms. Methicillin-resistant (Mcr) isolates from Melbourne (1982) and from Hobart (1986) were closely related to a methicillin-multiresistant S. aureus (MRSA) strain, ANS46, originally isolated in Melbourne in 1982 and studied extensively since. Methicillin-sensitive (Mcs) isolates were isolated concurrently with the Melbourne and Hobart Mcr isolates. These were found to be similar to induced Mcs variants of ANS46; these laboratory variants have lost approximately 40-70 kb of DNA carrying multiple resistance (R) determinants clustered around the mec gene. The Melbourne and Hobart Mcs isolates appear to be natural variants lacking this region of their chromosome. A clinical Mcr and Mcs pair of isolates differing only in the presence of an R-cluster near the mec gene were also isolated in Melbourne in 1990; these are not of the same clonal line as the earlier types from Melbourne and Hobart. These data suggest that insertion or deletion (or both) similar to that produced by known mutagens occurs in the mec region of the chromosome of MRSA in clinical populations under natural selective pressures; such processes may be important in the balance of resistant and sensitive staphylococci in hospitals and other clinical environments.

Australia↗

Analysis of an outbreak of variably methicillin-resistant Staphylococcus aureus with chromosomal RFLPs and mec region probes.

An outbreak of infections with multi-resistant Staphylococcus aureus with unusual methicillin resistance at a Melbourne hospital was investigated by examining restriction fragment length polymorphisms (RFLPs) of total DNA digested with the rare-cutting endonuclease SmaI. The polymorphisms were identified by pulsed-field gel electrophoresis (PFGE) and were analysed numerically to give quantitative estimates of genetic distances between isolates. The majority of the isolates were found to belong to one group, with only minor genetic differences between the isolates that showed varying resistance to methicillin, thereby suggesting development of resistant variants from one clonal type during the outbreak. These results were confirmed by DNA hybridisation analysis with specific resistance gene probes for parts of a multi-resistance gene cluster (including methicillin) in the chromosome. Analysis of the RFLP patterns of S. aureus isolates is potentially a useful procedure in clinical epidemiology.

Australia↗

Potential problems in the use of oligonucleotide probes for staphylococcal enterotoxin genes.

Oligonucleotide probes unique to the five major enterotoxin genes of Staphylococcus aureus were synthesized and used to detect DNA sequences homologous to these genes in 27 non-clinical isolates of Staph. aureus isolated from nasal swabs of 74 healthy human volunteers. Genomic DNA from all 27 isolates reacted with at least one of the probes. In a phenotypic assay for toxin production by a reverse passive latex agglutination test however, only 15 of the 27 isolates produced enterotoxin in culture. The results raise the possibility that a number of Staph. aureus isolates harbour DNA sequences that are apparently silent or mutant copies of the enterotoxin genes. This complicates the identification of enterotoxin producers by tests which depend on oligonucleotide or DNA hybridization.

Base Sequence↗

Lethal and mutational effects of solar and UV radiation on Staphylococcus aureus.

Strains of Staphylococcus aureus, an opportunistic pathogen commonly found on human skin, were exposed to sunlight and UV C radiation, and the lethal and mutational effects measured. Sunlight killed cells with an inactivation constant of 3 x 10(-5) per joule per square metre; UV C was much more lethal, giving an inactivation constant of approximately 0.1 per joule per square metre. Some strains tested showed a sensitivity to sunlight that was dependent on the growth phase of the cells, exponentially growing cells showing a greater sensitivity. Mutational effects of irradiation were measured by the appearance of mutants sensitive to methicillin following irradiation of a multiresistant strain. Mutants appeared at a frequency of 10(-3); this high frequency of mutation in the region of the mec gene has also been observed when multiresistant strains are subjected to nutritional or thermal stress. Mutants showed the same chromosomal alteration (seen in pulse-field gel electrophoresis of SmaI-digested DNA) whether induced by solar or UV C irradiation.

DNA Damage↗

Physical mapping of the mec region of an Australian methicillin-resistant Staphylococcus aureus lineage and a closely related American strain.

Methicillin-resistant (Mcr) staphylococci contain chromosomal DNA that is absent from Mcs cells. This extra DNA harbours the methicillin resistance determinant mec and often other resistance determinants. The mec region can differ substantially in structure among different isolates. We present studies on the mec region of a group of Staphylococcus aureus isolates prevalent in Australia and London. Southern hybridization analyses of a prototype Australian isolate, ANS46, and an isogenic Mcs deletion mutant, ANS62, allowed the physical map of the region to be extended to 55 kb. The DNA corresponding to the deletion, which includes mec and resistance determinants for mercury, cadmium (Cd) and tetracycline, amounted to 41 kb. It was bounded precisely at one end by the macrolides-lincosamides-streptogramin B (MLS)-resistance transposon, Tn554. Near the other end was an element with homology to Tn554, psi Tn554, which carried the Cdr determinant. The mec region of an American Mcr isolate, R35, was found to be virtually the same as that of ANS46, except that it lacked Tn554. Another class of American Mcr isolates, prevalent since 1987, differs markedly from ANS46 in mec region organization. However, this other American class also contains an insertion of Tn554 in the mec region, and the attachment site for this insertion was found to have significant homology to attachment sites for the Tn554 and psi Tn554 insertions in the mec region of the Australian strain. These results suggest possible roles of Tn554 and Tn554-like elements in the evolutionary variation of the mec region.

Australia↗

The effect of muscle vibration on human position sense during movements controlled by lengthening muscle contraction.

Muscle vibration studies suggest that during voluntary movement limb position is coded by muscle spindle information derived from the lengthening, antagonist muscle. However, these investigations have been limited to movements controlled by shortening contractions. This study further examined this property of kinesthesia during movements controlled by lengthening contraction. Subjects performed a horizontal flexion of the right forearm to a mechanical stop randomly positioned at 30, 50 and 70 degrees from the starting position. The movement was performed against a flexor load (1 kg) requiring contraction of the triceps muscle. Vision was occluded and movements were performed under three conditions: no vibration, vibration of the right biceps and vibration of the right triceps. The perceived position of the right forearm was assessed by instructing subjects to simultaneously match the right limb position with the left limb. Vibration of the shortening biceps muscle had no effect on limb matching accuracy. However, triceps vibration resulted in significant overestimation of the vibrated limb position (10-13 degrees). The variability in movement distance was uninfluenced by muscle vibration. During movements controlled by lengthening contraction, there is a concurrent gamma dynamic fusimotor input that would enhance primary afferent discharge. Despite this additional regulating input to the muscle spindle, it appears that muscle spindle information from the lengthening muscle is important for the accurate perception of limb movement and/or position.

Adult↗

Epidemiological analysis of a methicillin-resistant Staphylococcus aureus outbreak using restriction fragment length polymorphisms of genomic DNA.

The genomic DNA of 58 isolates of methicillin-resistant Staphylococcus aureus (MRSA) obtained during an infection outbreak at two major Canberra hospitals was analysed for restriction fragment length polymorphism (RFLP) by digestion with the endonuclease SmaI and resolution of the fragments by pulsed-field gel electrophoresis. Based on the fraction of common fragments generated by the endonuclease, DNA similarities among the isolates were estimated. Distance matrix analysis showed that the MRSA isolates could be divided into two major clusters (RFLP types I and II) and one minor one (type 46). A fourth group of miscellaneous isolates was found to be heterogeneous in terms of DNA sequence similarity. The epidemiological data indicated that RFLP type I was most common in the intensive care units in the two hospitals, with particular subtypes of RFLP type I concentrated in individual units. RFLP type II and the miscellaneous group were more generally distributed. Type 46 isolates appear to be related to a group which was present in epidemics in Melbourne hospitals in the early 1980s. Using the standard phage set, the RFLP type I group was largely untypable. However, type II isolates were all phage typable, with a shared susceptibility to phages 29/85/95/90; type 46 isolates had a shared susceptibility to phages 85/90. The miscellaneous isolates were of variable phage types.

Australia↗

Bacteriophage 604: a marker phage for multi-resistant Staphylococcus aureus in Australia.

Of 28 multi-resistant isolates of Staphylococcus aureus collected during 1986 from hospitals in major cities around Australia, 27 were found to contain the same prophage (denoted phage 604). Hospital isolates carrying three or fewer resistance markers, and community isolates carrying one or no resistance markers, did not carry this prophage. Phage 604 does not confer antibiotic resistance on its lysogens, nor does it increase virulence in chick embryo assays. Phage 604 appears to be a correlate of antibiotic multi-resistance in S. aureus in Australia, and may provide a molecular marker for incipiently epidemic strains of this bacterium in Australian hospitals.

Animals↗

Induced deletions within a cluster of resistance genes in the mec region of the chromosome of Staphylococcus aureus.

Variants of a methicillin-resistant Staphylococcus aureus showing loss of or reduced resistance to the antibiotic were isolated at frequencies of 0.1-100% from cultures which had been starved, grown at elevated temperature, or given small doses of UV radiation. Three types of variant were identified on the basis of population distribution of resistance to the antibiotic, and field-inversion gel electrophoresis of digests of the chromosome cut with the rare-cutting restriction endonuclease SmaI. Type I variants are methicillin-sensitive and have a deletion in the mec region of the chromosome. Type II variants have reduced methicillin resistance and rearranged DNA elsewhere in the chromosome. Type II variants show reduced methicillin resistance and no detectable change in the chromosome. Type I deletions were mapped using cloned fragments from the mec region. In 13 of the 16 independently isolated deletion mutants, one of the deletion endpoints appears to correlate with the positions of insertion sequences or transposons found in this region of the staphylococcal chromosome.

Chromosome Deletion↗

The expression in Staphylococcus aureus of cloned DNA encoding methicillin resistance.

A 4 kb fragment of chromosomal DNA was cloned from a clinical strain of methicillin-resistant Staphylococcus aureus. It comprises part of a section of the chromosome that was lost when the strain was cured of resistance to methicillin and to other antimicrobial agents. The fragment mediates an increased level of methicillin resistance when inserted into a shuttle vector and transformed back into the sensitive strain generated when the original DNA was deleted.

Chromosomes, Bacterial↗

Mercury and tetracycline resistance genes and flanking repeats associated with methicillin resistance on the chromosome of Staphylococcus aureus.

Sections of a cloned 27 kb segment of chromosomal DNA, associated with resistance to four antimicrobial agents in a clinical isolate of methicillin-resistant Staphylococcus aureus (MRSA), were tested for their ability to determine resistance when transformed into a sensitive laboratory strain of S. aureus. This was achieved by inserting the sections into a newly constructed shuttle vector, amplifying the recombinant DNA in E. coli, and transforming protoplasts of the sensitive S. aureus strain. Two sections of the cloned DNA were found to determine resistance separately to mercuric ion and to tetracycline, in both S. aureus and Escherichia coli.

Chromosomes, Bacterial↗

Molecular relatedness of Staphylococcus aureus typing phages measured by DNA hybridization and by high resolution thermal denaturation analysis.

Fifteen bacteriophages representative of the serological and lytic groups of the International Typing Set for Staphylococcus aureus were examined for genomic homology by DNA hybridization and by analysis of high resolution thermal denaturation profiles. Phages 11 and 80 alpha, not part of the set, were also examined. DNA homology measured by filter hybridization showed values ranging from near zero to 88 per cent in pair-wise comparisons. Cluster analysis of these data by standard numerical taxonomical methods yielded clusters which closely reflect the subdivision of the international set on the basis of serological reactions. High resolution thermal denaturation analysis yielded characteristic profiles for each phage DNA, with members closely related by hybridization analysis showing only minor differences. Quantitative analysis of the extent of overlap of these profiles generated relational values which were subjected to the same numerical taxonomic analysis as for the DNA hybridization data. The resultant dendrogram was qualitatively different only in minor respects from that derived from the hybridization analyses, but quantitatively homology was greater by 80 per cent or more for the DNAs which were least related according to the hybridization analyses. This upward shift in measured homology appears to reflect the similar base composition of the DNAs from these phages. Statistical comparison of the homology data obtained by the two methods showed them to be significantly correlated. These results indicate that the International Typing Set consists of phages which all appear to be related to a greater or lesser extent. If, as the history of the collection of these phages indicates, they are a random sample of aureophages, then this group of phages may represent a common genetic pool within which recombination, mutation, and genome rearrangement occur to generate unique individual phages.

DNA, Viral↗