Diagnostic value of platelet counts in mammals.
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Animal Diseases↗
Biomedical subjects
P M Bennett
Publications and source records attributed to P M Bennett.
Calcium regulated thin filaments from molluscan catch muscles contain a caldesmon-like regulatory protein.
The thin filaments of the anterior byssus retractor muscle of the edible mussel Mytilus and the transluscent and opaque adductors of the oyster Crassostrea have been isolated and their properties investigated. We find that the thin filaments from all three muscles can activate skeletal muscle myosin ATPase in the presence of calcium but that the activity is inhibited in its absence. The filaments contain a protein which interacts with antibodies to vertebrate smooth muscle caldesmon on immunoblots. The antibodies relieve the inhibition of the thin-filament-activated myosin MgATPase. They can also bundle the thin filaments. We conclude that a caldesmon-like protein is present in molluscan muscle. As in the vertebrate smooth muscle, it could act as part of a control mechanism in addition to the myosin regulatory system. Vertebrate smooth muscle caldesmon can crosslink actin and myosin and it has been suggested that it may in this way contribute to the latch state. A similar interaction may be involved in the catch mechanism in molluscan muscle.
Actin Cytoskeleton↗
Broad spectrum beta-lactamases of Citrobacter diversus.
Citrobacter diversus NF85 produced a chromosomal beta-lactamase that was induced by a variety of beta-lactam antibiotics. Two major forms of the enzyme, with isoelectric points (pI's) of 5.7 and 6.2, were found in crude cell extracts. Derepressed mutants of NF85, generated by nitrosoguanidine treatment, displayed different levels of beta-lactamase expression to the parent strain and had different patterns of resistance to a range of beta-lactam antibiotics. Those mutants of NF85 that were totally derepressed, expressing high, constitutive levels of enzyme, were found to have an additional beta-lactamase activity with a pI of 6.8.
Anti-Bacterial Agents↗
Induction of the Citrobacter freundii group I beta-lactamase in Escherichia coli is not dependent on entry of beta-lactam into the cytoplasm.
Induction of the cloned ampC Citrobacter freundii beta-lactamase in Escherichia coli by 6-aminopenicillanic acid was prevented by periplasmic TEM beta-lactamase in the same cell. In contrast, cytoplasmic TEM beta-lactamase did not prevent induction. This result implies that entry of the beta-lactam into the cytoplasm is not necessary for induction of ampC.
Anti-Bacterial Agents↗
Paracrystals of myosin rod.
To help understand the packing of myosin tails in the backbone of the vertebrate striated muscle thick filament, paracrystals of myosin rod, a proteolytic fragment corresponding to the whole myosin tail, have been examined by electron microscopy and image analysis. Two types of paracrystal were observed. Type I paracrystals were similar to those seen by Moos et al. (1975; J. molec. Biol. 97, 1-9). These showed a 14-nm axial repeat, but yielded no other structural information. Type II paracrystals were long, flexible ribbons with more regularity. When negatively stained they exhibited a weak 43-nm axial striation and appeared to be composed of a layer of narrow filaments. Optical diffraction showed that the paracrystals had a rectangular unit cell of dimensions 43 nm axially and 12.4 nm laterally. Transverse sections indicated a paracrystal depth similar to the lateral dimension of the unit cell. Each unit cell contained two filaments arranged antiparallel and related by a two-fold screw axis perpendicular to the length, and in the plane of the ribbon. The filaments probably consist of parallel rod molecules related by axial displacements of 43 nm and higher multiples of 43 nm. The nature of these paracrystals indicates that the myosin tail alone can form structures like thick filament subfilaments. Their structure, based on distinguishable parallel and antiparallel rod interactions, was sensitive to pH and divalent cations in a similar way to the ionic effects on the structure of thick filaments. This behaviour suggests that some of the interactions present in the paracrystal are the same as those in the thick filament.
Animals↗
The 'catch' mechanism in molluscan muscle: an electron microscopy study of freeze-substituted anterior byssus retractor muscle of Mytilus edulis.
A method for quick-freezing muscles while observing their mechanical properties until the moment of freezing is described. This method was used to freeze the anterior byssus retractor muscle (ABRM) of Mytilus edulis. Intact muscle in the presence of sucrose as a cryoprotectant was freeze-substituted in acetone, fixed and embedded for electron microscopy. ABRM was frozen in a number of mechanical states including 'catch', the state of high passive tension particularly associated with some molluscan muscles. Transverse sections were examined to determine the distribution of filaments in the muscle cells. In the relaxed muscle thick and thin filaments are fairly randomly distributed. Groups of thin filaments and of thick filaments are often seen, and there is no obvious association between the two types of filaments. In contrast, in rigor muscles, both glycerol-extracted and intact, most of the thin filaments were found to lie in rings or rosettes around the thick filaments. In some places bridges between thick and thin filaments could be distinguished. In actively contracting muscle (phasic contraction) the appearance is intermediate between that of the relaxed and rigor muscles. Many thick filaments are surrounded by rosettes of thin filaments but many of the thin filaments are grouped and have no connections with thick filaments. The 'catch' state, left after a period of tonic contraction, is similar in its distribution of thick and thin filaments to the active state, many of the thin filaments lying between the thick. Frequently thick and thin filaments seem to be closer together than in other states of the muscle where a pronounced exclusion zone is present around the thick filaments. There is no evidence for association between the thick filaments. The different distribution of thin filaments in the different states is consistent with the previously described X-ray diffraction data if it is assumed that most of the contribution to the equatorial reflection at 12 nm comes from the groups of thin filaments. Our data support a model for catch in which there is a change in the association between thick and thin filaments, rather than one in which thick filaments are clumped.
Actin Cytoskeleton↗
Characterization of Pseudomonas mercury-resistance transposon Tn502, which has a preferred insertion site in RP1.
Tn502mer differs in size and restriction map from the well-characterized Tn501mer. It also differs in its preferential and high-frequency insertion into the 6 kb PstI-C region of RP1. The affinity for this region is perpetuated in pVS76, a clone of RP1 PstI-C in pBR322. Restriction mapping of independent pVS76::Tn502 derivatives revealed that Tn502 inserted at the same site (or small region) in PstI-C corresponding to the 35 kb coordinate in RP1. Insertion occurred in both orientations, but one was preferred. When PstI-C was deleted from RP1, acquisition of Tn502 was reduced and the sites of insertion randomized.
Binding Sites↗
Penicillin-binding protein 2 is required for induction of the Citrobacter freundii class I chromosomal beta-lactamase in Escherichia coli.
Involvement of Escherichia coli penicillin-binding proteins (PBPs) in induction of the cloned ampC Citrobacter freundii beta-lactamase by 6-aminopenicillanic acid was investigated. The enzyme was not inducible at 42 degrees C in a mutant thermosensitive for expression of PBP 2. The results imply that PBP 2 is involved in the process leading to induction of ampC.
Acyltransferases↗
Cloning and characterization of an AAC(6') gene from Serratia marcescens.
A gene encoding AAC(6') mediated resistance to netilmicin and amikacin was cloned from the chromosome of a nosocomial strain of Serratia marcescens into the BamHI site of pACYC184. The resistance determinant was localized to a sequence of approximately 1 kb. A 490 base pair BamHI-PvuII restriction fragment from the cloned DNA was used as a probe in Southern hybridization studies. Homology with a 920 base pair fragment of PvuII digested S. marcescens DNA from AAC(6')-producing strains was observed. No homology was detected with aminoglycoside sensitive S. marcescens or Escherichia coli.
Acetyltransferases↗
Monitoring for the development of antimicrobial resistance during the use of olaquindox as a feed additive on commercial pig farms.
Since 1982, when olaquindox was introduced as a pig-feed additive in the UK, about 12 commercial farms in Suffolk have been monitored annually to check for the possible emergence of resistance to olaquindox and chloramphenicol among the coliform flora of the pigs and their environment. In spite of the sampling variability and the impossibility of controlling the use of feed additives and management on the farms, the overall results obtained were consistent and, it is suggested, the method is widely applicable. A steady, albeit low, increasing incidence and level of resistance to olaquindox was recorded (1982-1984) on farms using it and, to a lesser degree, on neighbouring farms that did not. No significant increase in the level of chloramphenicol resistance was observed. Genetical studies on a selection of olaquindox-resistant isolates suggested that the genes determining resistance were likely to be borne on the chromosome.
Animal Feed↗
Inhibition of Providencia stuartii cell envelope enzymes by chlorhexidine.
The possibility that chlorhexidine is a specific inhibitor of membrane bound bacterial adenosine triphosphatase (ATPase) was addressed. The in-vitro susceptibilities of several Providencia stuartii cell envelope enzymes, including ATPase, to chlorhexidine were compared. The following concentrations of chlorhexidine were required to cause 50% inhibition of enzyme activity in preparations from chlorhexidine-sensitive strains (MIC 50 mg chlorhexidine/l): ATPase (160 mg/l), succinic dehydrogenase (greater than 300 mg/l), penicillin binding protein 7 (300 mg/l) and beta-lactamase (45 mg/l). Fifty per cent inhibition of the ATPase from a chlorhexidine-resistant strain (MIC 1600 mg/l) was achieved at an in-vitro concentration of 225 mg chlorhexidine/l. Our observations do not support the suggestion that bacterial membrane-bound ATPases are specific targets for chlorhexidine.
Adenosine Triphosphatases↗
Citrobacter diversus ULA27 produces two forms of a chromosomal beta-lactamase.
A multi-resistant clinical isolate of Citrobacter diversus, particularly resistant to penicillins and cephalosporins, has been studied. The strain produced constitutively high levels of beta-lactamase which efficiently hydrolysed cephaloridine but not ampicillin. The apparent Km value for cephaloridine (125 +/- 25 microM) was within the range usually observed for beta-lactamases in similar bacteria. The strain possessed two forms of the enzyme as indicated by isoelectric focusing patterns, HPLC analysis and two pH optima. SDS-PAGE analysis indicated a single protein responsible for both forms. The strain possessed a single plasmid, as revealed by agarose gel electrophoresis, but beta-lactamase production was not plasmid encoded. The available data indicate the beta-lactamase(s) of C. diversus ULA27 to be chromosomally encoded.
Anti-Bacterial Agents↗
The R46 site-specific recombination system is a homologue of the Tn3 and gamma delta (Tn1000) cointegrate resolution system.
The nucleotide sequence of the R46 site-specific recombination system has been determined. The organization of the recombination gene (perR46) and the site at which it acts (per site), together with the extensive sequence homology displayed with the tnpR genes and res sites of the transposons Tn3 and gamma delta (Tn1000), suggests that they have been derived from a Tn3-like element. These site-specific recombination functions of R46 play a role in plasmid maintenance.
Base Sequence↗
Genetic basis of the spread of antibiotic resistance genes.
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Anti-Bacterial Agents↗
An ultra-rapid method for the study of antibiotic resistance plasmids.
A rapid method of isolating plasmid DNA has been developed from that of Birnboim & Doly (1979). This method allows large numbers of strains to be examined, and can be employed to isolate DNA from members of the Enterobacteriaceae, Pseudomonas aeruginosa, Staphylococcus aureus, (including methicillin-resistant strains) and coagulase-negative staphylococci. Plasmids of widely differing sizes are amenable to isolation by this technique, which yields DNA of sufficient purity to allow restriction endonuclease and homoduplex analysis.
Bacteria↗
Evolution of R plasmids by replicon fusion.
The occurrence of plasmid-plasmid fusions in nature and their possible influence on the spread of bacterial drug resistance is considered. Three ways in which replicon fusions can be formed are described; specifically fusions mediated by IS elements, "one-ended transposition" systems derived from Tn3 and like transposons, and pUB2380, a prototype for a new type of transposition. Non-transposition based systems which mediate replicon fusion are also briefly considered.
Biological Evolution↗
Do plasmids influence the survival of bacteria?
Bacterial plasmids may contribute to the fitness of a particular bacterial clone to survive in a specific environment. In many cases, however, the determinants that confer the advantage are unknown. The advantages to be gained from plasmid carriage appear to outweigh potential disadvantages imposed by an increased metabolic demand.
Bacteria↗
Location of the site-specific recombination system of R46: a function necessary for plasmid maintenance.
The R46 site-specific recombination system comprises a per (plasmid-encoded recombinase) gene and a site at which the gene product acts, the per site. The two functions have been cloned into pACYC184. They are encoded by sequences within a region of approximately 2 kb on the R46 genome. These R46 sequences are closely related to the site-specific recombination systems of the ampicillin resistance transposons collectively designated TnA. The R46 per function is interchangeable with the tnpR gene product of TnA. Both enzymes can mediate recombination between the related res and per sites in R46::TnA recombinant plasmids to generate site-specific deletions and inversions. Similar DNA rearrangements occur when TnA inserts into pACYC184 derivatives carrying the cloned R46 per functions. Carriage of this site-specific recombination system contributes to the stable maintenance of R46. By converting plasmid dimers to monomers the R46 per functions help to ensure equal partitioning at cell division.
Cloning, Molecular↗