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Biomedical subjects

R J Wilson

Publications and source records attributed to R J Wilson.

At least 37 records · Page 2Linked to original sources
PubMed2000

Bacterial orthologues indicate the malarial plastid gene ycf24 is essential.

ycf24 is a well conserved gene found in all major groups of bacteria, as well as on red algal plastid genomes and the vestigal plastid genome of apicomplexan pathogens like the malaria parasite Plasmodium falciparum (ORF470). Some database annotations describe Ycf24 as an ABC transporter subunit, but we find the level of significance is low. To investigate ycf24's function we disrupted it in the cyanobacterium Synechocystis sp., strain PCC6803 which has a multi-copy genome. This showed ycf24 is essential, partial loss producing a terminal phenotype of chlorosis, reduced cell size, loss of DNA, and a striking arrest in cytokinesis. Attempts to disrupt the single copy of ycf24 in E. coli failed to give stable transformants. When Ycf24 was over-expressed in E. coli as a soluble fusion protein, it localized mostly as a band on either side of the nucleoid and nucleoid partitioning was aberrant. We propose the relict plastid organelle of apicomplexans retains its capacity for protein synthesis because Ycf24 is essential.

ATP-Binding Cassette Transporters↗
PubMed2000

Actinobacillus suis infection in pigs in Australia.

This is the first report of the isolation of Actinobacillus suis in association with significant disease among preweaned pigs in Australia. Sudden deaths occurred in preweaned piglets at one facility and enlarged joints, particularly the stifles and tarsi, occurred at another. Isolates with the biochemical phenotype and apx genotype profile of A suis were cultured from affected piglets. Both facilities were of high health status and one had undergone a recent depopulation, disinfection and sow repopulation process. Reviewing initial reports of A suis disease overseas, it is apparent that outbreaks occurred sporadically in various locations, with disease occurring among the herds involved for some months only, before development of immunity. A suis disease is now considered an emerging disease on high health status farms.

Actinobacillus↗
PubMed2000

Evolution of air-breathing and central CO(2)/H(+) respiratory chemosensitivity: new insights from an old fish?

While little is known of the origin of air-breathing in vertebrates, primitive air breathers can be found among extant lobe-finned (Sarcopterygii) and ray-finned (Actinopterygii) fish. The descendents of Sarcopterygii, the tetrapods, generate lung ventilation using a central pattern generator, the activity of which is modulated by central and peripheral CO(2)/H(+) chemoreception. Air-breathing in Actinopterygii, in contrast, has been considered a 'reflexive' behaviour with little evidence for central CO(2)/H(+) respiratory chemoreceptors. Here, we describe experiments using an in vitro brainstem preparation of a primitive air-breathing actinopterygian, the longnose gar Lepisosteus osseus. Our data suggest (i) that gill and air-breathing motor patterns can be produced autonomously by the isolated brainstem, and (ii) that the frequency of the air-breathing motor pattern is increased by hypercarbia. These results are the first evidence consistent with the presence of an air-breathing central pattern generator with central CO(2)/H(+) respiratory chemosensitivity in any primitive actinopterygian fish. We speculate that the origin of the central neuronal controller for air-breathing preceded the divergence of the sarcopterygian and actinopterygian lineages and dates back to a common air-breathing ancestor.

Animals↗
PubMed2000

Segmental control of midbody peristalsis during the consummatory phase of feeding in the medicinal leech, Hirudo medicinalis.

The ingestive behavior of the medicinal leech includes peristalsis-like movements of the midbody that are organized into both rostral-to-caudal and caudal-to-rostral waves (C. M. Lent, K. H. Fliegner, E. Freedman, & M. H. Dickenson, 1988). The neuronal control of this behavior is unknown. Using surgical manipulations and electromyograms, the authors show that (a) the head and tail ganglia are not necessary for this behavior; (b) the circuit is distributed, with components reiterated along the length of the leech; (c) excitatory signals transmitted from rostral segments via the nerve cord can initiate peristalsis in "empty" caudal segments; (d) inhibitory signals from caudal segments limit the frequency of peristalsis; and (e) stretch of the gut and/or body wall is sufficient to produce peristalsis in the absence of heat or chemical cues. These results are compared with peristalsis in the digestive tract of mammals. The leech may be a good model for studying peristalsis-like behaviors at the cellular level.

Animals↗
PubMed1999

Efficacy of a low volume recirculating superfusion chamber for long term administration of expensive drugs and dyes.

We have characterized the efficacy of a low volume (2 ml) two-compartment experimental chamber in which a gas inflow equilibrates and recirculates the bathing fluid. This type of chamber is suitable for experiments employing en bloc preparations that require the administration of expensive molecular probes. The fluid in the chamber is pumped from a compartment holding the preparation to an elevated reservoir compartment using gas bubbles. The fluid returns via gravity along a different path. The flow rate of superfusate in the chamber was 30 ml min(-1). To determine the effectiveness of the chamber in dissolving gas, we filled the chamber with bicarbonate-buffered physiological saline and measured pH and P(O(2)) with ion-selective and Clark-style microelectrodes. Steady state values of pH and P(O(2)) in the chamber were almost identical to those in an external tonometer bubbled vigorously with the same gas mixture. When CO(2) was increased from 2 to 4.4%, the chamber pH fell with a time constant of 56 s (about twice that of the tonometer). To determine the effectiveness of gas exchange between a brain preparation and the fluid in the chamber we measured pH and P(O(2)) depth profiles of the in vitro tadpole brainstem. We found virtually no unstirred layer owing to excellent mixing and the high flow created by the recirculating mechanism. We demonstrate that despite the high flow rates, preparations are mechanically stable allowing intracellular electrophysiological recordings.

Animals↗
PubMed1999

Protein synthesis in the plastid of Plasmodium falciparum.

The plastid organelle of malarial and other apicomplexan parasites contains ribosome-like particles as well as a genome dedicated largely to specifying components of a protein expression system. We have identified plastid ribosomes using hybridization studies and show that in erythrocytic stages of Plasmodium falciparum a subset of polysomes carries plastid-specified rRNAs and mRNA, supporting the idea that protein synthesis is active in the plastid.

Animals↗
PubMed1999

Antibiotic inhibitors of organellar protein synthesis in Plasmodium falciparum.

Elongation factor Tu (EF-Tu) is encoded by the tuf gene of the plastid organelle of the malaria parasite Plasmodium falciparum. A range of structurally unrelated inhibitors of this GTP-dependent translation factor was shown to have antimalarial activity in blood cultures. The most active was the cyclic thiazolyl peptide amythiamicin A with an IC50 = 0.01 microM. Demonstrable complexes were formed in vitro between a recombinant version of P. falciparum EF-Tu(pl) and inhibitors that bind to different sites on EF-Tu; these included the antibiotics kirromycin, GE2270A and enacyloxin IIa.

Animals↗
PubMed1999

Emotional congruence in sexual offenders against children.

The tendency of some sexual offenders against children to display an exaggerated cognitive and emotional affiliation with childhood has been labeled "emotional congruence" by Finkelhor (1984) and as overidentification with childhood by others. This study consisted of an evaluation of emotional congruence in 194 men--homosexual and heterosexual pedophiles (N's = 41 and 31), heterosexual nonpedophilic incest offenders (N = 62), sexual offenders against women (N = 27), and nonsexual offender criminals (N = 33). A revised version of the Child Identification Scale (CIS-R) was used in combination with a semistructured interview (SSI) also intended to measure the construct. Maximum-likelihood factor analysis of the CIS-R generated eight factors. All subjects, particularly the nonsexual offenders, scored high on a factor indicating a wish to restart their lives. With respect to emotional congruence, results showed that only the homosexual pedophiles' scores indicated a preference for interacting with children on the child's level, while the incest offenders seemed to prefer to elevate their victims to adult status rather than fixating on the child role themselves. The group of heterosexual pedophiles, however, seemed to be motivated more by sexual gratification than by an emotional or relationship interest in their victims or children.

Adult↗
PubMed1998

Actomyosin motor in the merozoite of the malaria parasite, Plasmodium falciparum: implications for red cell invasion.

The genome of the malaria parasite, Plasmodium falciparum, contains a myosin gene sequence, which bears a close homology to one of the myosin genes found in another apicomplexan parasite, Toxoplasma gondii. A polyclonal antibody was generated against an expressed polypeptide of molecular mass 27,000, based on part of the deduced sequence of this myosin. The antibody reacted with the cognate antigen and with a component of the total parasite protein on immunoblots, but not with vertebrate striated or smooth muscle myosins. It did, however, recognise two components in the cellular protein of Toxoplasma gondii. The antibody was used to investigate stage-specificity of expression of the myosin (here designated Pf-myo1) in P. falciparum. The results showed that the protein is synthesised in mature schizonts and is present in merozoites, but vanishes after the parasite enters the red cell. Pf-myo1 was found to be largely, though not entirely, associated with the particulate parasite cell fraction and is thus presumably mainly membrane bound. It was not solubilised by media that would be expected to dissociate actomyosin or myosin filaments, or by non-ionic detergent. Immunofluorescence revealed that in the merozoite and mature schizont Pf-myo1 is predominantly located around the periphery of the cell. Immuno-gold electron microscopy also showed the presence of the myosin around almost the entire parasite periphery, and especially in the region surrounding the apical prominence. Labelling was concentrated under the plasma membrane but was not seen in the apical prominence itself. This suggests that Pf-myo1 is associated with the plasma membrane or with the outer membrane of the subplasmalemmal cisterna, which forms a lining to the plasma membrane, with a gap at the apical prominence. The results lead to a conjectural model of the invasion mechanism.

Actomyosin↗
PubMed1997

A plastid of probable green algal origin in Apicomplexan parasites.

Protozoan parasites of the phylum Apicomplexa contain three genetic elements: the nuclear and mitochondrial genomes characteristic of virtually all eukaryotic cells and a 35-kilobase circular extrachromosomal DNA. In situ hybridization techniques were used to localize the 35-kilobase DNA of Toxoplasma gondii to a discrete organelle surrounded by four membranes. Phylogenetic analysis of the tufA gene encoded by the 35-kilobase genomes of coccidians T. gondii and Eimeria tenella and the malaria parasite Plasmodium falciparum grouped this organellar genome with cyanobacteria and plastids, showing consistent clustering with green algal plastids. Taken together, these observations indicate that the Apicomplexa acquired a plastid by secondary endosymbiosis, probably from a green alga.

Animals↗
PubMed1997

Predicting survival from out-of-hospital cardiac arrest: a multivariate analysis.

From 954 attempts to resuscitate patients from out-of-hospital cardiac arrest two datasets were derived, namely 861 cases of cardiac arrest and 906 cases of either cardiac or primary respiratory arrest. For each dataset, multivariate analysis was performed by fitting a number of explanatory variables with respect to the outcomes of admission to hospital and discharge home in logistic regression models. There were numerous interactions between these variables. Being conscious at the time of the arrival of the ambulance crew and subsequently having cardiac arrest strongly predicted survival, as did both the presence of a witness to the arrest and the initiation of cardiopulmonary resuscitation (CPR) by a bystander; this latter effect was a marker for early CPR. The strongest predictor of a poor outcome was delay to CPR or delay to advanced cardiac life support.

Age Distribution↗
PubMed1997

Outcome of out-of-hospital cardiorespiratory arrest in south Glamorgan.

During 138 weeks an emergency medical service (EMS) of mixed skill-level attempted to resuscitate 954 patients from prehospital cardiac arrest (883 attempts per million population per year); 75% of the arrests were of cardiac cause. This paper is one of the first analyses from europe to use the 'Utstein template' to report outcomes of such arrests. In cases where an arrest rhythm could be recorded, 38.4% were ventricular fibrillation (VF), 45.5% were asystolic, and the remainder were either electromechanical dissociation or respiratory arrests. Using univariate analysis factors associated with a greater likelihood of survival include the presence of a witness, bystander-initiated cardiopulmonary resuscitation (CPR), early CPR and VF as the arrest rhythm. Twenty of 155 cases (13%) survived where VF arrest was witnessed by non-EMS personnel.

Aged↗
PubMed1997

Extrachromosomal DNA in the Apicomplexa.

Malaria and related apicomplexan parasites have two highly conserved organellar genomes: one is of plastid (pl) origin, and the other is mitochondrial (mt). The organization of both organellar DNA molecules from the human malaria parasite Plasmodium falciparum has been determined, and they have been shown to be tightly packed with genes. The 35-kb circular DNA is the smallest known vestigial plastid genome and is presumed to be functional. All but two of its recognized genes are involved with genetic expression: one of the two encodes a member of the clp family of molecular chaperones, and the other encodes a conserved protein of unknown function found both in algal plastids and in eubacterial genomes. The possible evolutionary source and intracellular location of the plDNA are discussed. The 6-kb tandemly repeated mt genome is the smallest known and codes for only three proteins (cytochrome b and two subunits of cytochrome oxidase) as well as two bizarrely fragmented rRNAs. The organization of the mt genome differs somewhat among genera. The mtDNA sequence provides information not otherwise available about the structure of apicomplexan cytochrome b as well as the unusually fragmented rRNAs. The malarial mtDNA has a phage-like replication mechanism and undergoes extensive recombination like the mtDNA of some other lower eukaryotes.

Animals↗
PubMed1996

A model of the hydrostatic skeleton of the leech.

A mathematical model of the hydrostatic skeleton of the leech has been developed to predict the shape of and internal pressure within the animal in response to a given pattern of motor neuron activity in different behaviors. The model incorporates experimental data on: the dimensions of the animal at behavioral extremes, the passive properties of the tissues, the active length-tension behavior of the muscles in response to neural activation, the relations between firing frequencies and forces developed by the muscles. The model is based on three general assumptions: (i) the cross-sectional geometry of each segment is elliptical, (ii) the volume of each segment remains constant during movement, (iii) the shape of the animal reflects dimensions that minimize the total potential energy. Presently the model is implemented to simulate the vermiform elongation of the leech, predicting the shape and the pressure changes during behavior. The results are in good agreement with the experimental measurements. The pattern of motor neuronal activity was determined by the known intersegmental travel time and estimated delay time between relaxation of the longitudinal muscles and the activation of the circular muscles. The anesthetized state of the leech was taken as the reference state for the model in which the active and passive stresses are zero.

Animals↗
PubMed1996

Complete gene map of the plastid-like DNA of the malaria parasite Plasmodium falciparum.

Malaria parasites, and other parasitic protists of the Phylum Apicomplexa, carry a plastid-like genome with greatly reduced sequence complexity. This 35 kb DNA circle resembles the plastid DNA of non-photosynthetic plants, encoding almost exclusively components involved in gene expression. The complete gene map described here includes genes for duplicated large and small subunit rRNAs, 25 species of tRNA, three subunits of a eubacterial RNA polymerase, 17 ribosomal proteins, and a translation elongation factor. In addition, it codes for an unusual member of the Clp family of chaperones, as well as an open reading frame of unknown function found in red algal plastids. Transcription is polycistronic. This plastid-like DNA molecule is conserved in several genera of apicomplexans and is conjectured to have been acquired by an early progenitor of the Phylum by secondary endosymbiosis. The function of the organelle (plastid) carrying this DNA remains obscure, but appears to be specified by genes transferred to the nucleus.

Amino Acid Sequence↗