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Biomedical subjects

J Stephen

Publications and source records attributed to J Stephen.

At least 37 records · Page 2Linked to original sources
PubMed1995

Biological and genetic characterization of TnphoA mutants of Salmonella typhimurium TML in the context of gastroenteritis.

TnphoA transposon insertion mutants of phoN-negative derivatives of Salmonella typhimurium TML (of human gastroenteritic origin) were selected by growing mutagenized recipient bacteria under a variety of growth conditions. Ninety-seven individual mutants, which expressed alkaline phosphatase, were collected and tested for their ability to invade HEp-2 cells. Seven smooth mutants had a reduced ability to invade HEp-2 cells, and three smooth mutants were consistently more invasive than their corresponding parental strains. One rough mutant was of similar invasiveness and two were of reduced invasiveness when compared with that of parental strains. The seven smooth hypoinvasive mutants, the three smooth hyperinvasive mutants, and the three rough mutant strains were tested for their abilities to invade ileal enterocytes by the rabbit ileal invasion assay described previously (3). All smooth mutants exhibited parental levels of invasiveness. The rough mutants were hypoinvasive in the rabbit ileal invasion assay. The HEp-2 system is therefore not a good predictor of behavior in gut tissue in this model. DNA sequences flanking the transposon were determined for five mutants which were hypoinvasive in the HEp-2 cell assay. The mutations were found to be insertions in two previously identified invasion genes, invG and invH, and in a gene not normally associated with invasion, pagC. These observations lead one to be cautious in the interpretation of the biological significance of data obtained from invasion of tissue culture monolayers when extrapolated to gut tissue.

Alkaline Phosphatase↗
PubMed1995

The East York Health Needs Study. I: Prevalence of DSM-III-R psychiatric disorder in a sample of Canadian women.

BACKGROUND: This study reports the prevalence of psychiatric disorder in women from a Canadian community. The GHQ and the CES-D were compared for their utility. METHOD: A thousand women over the age of 18 were mailed the GHQ and the CES-D. Our return rate was 44.4%; 24% were personally interviewed by interviews blind to screening information. The CIDI was used to establish DSM-III-R diagnoses. Four versions of the GHQ and one version of the CES-D were calibrated against the CIDI. RESULTS: The prevalence of general psychiatric disorder was estimated as between 15% and 19%, anxiety disorders between 10% and 13%, and depression occurring with anxiety between 3% and 4%. The calibrated GHQ was the most reliable instrument. CONCLUSIONS: Prevalence of DSM-III-R psychiatric disorder can be reliably determined with the calibrated GHQ. Anxiety disorders are most prevalent in this community, and were best detected using calibrated versions of the longer form GHQ.

Adolescent↗
PubMed1995

The cochlear lesion in experimental bacterial meningitis of the rabbit.

Sensorineural hearing loss was studied in a rabbit model of experimental bacterial meningitis using electrophysiological and ultrastructural techniques. Hearing impairment was monitored by auditory brain-stem evoked responses (ABERs) and concomitant structural lesions were identified by both transmission (TEM) and scanning (SEM) electron microscopy. Meningitis was induced by intra-cerebrospinal fluid injection of either Escherichia coli (strain 2073 and type K-12) or Haemophilus influenzae type b. Auditory loss of approximately equal to 10 dB occurred in all rabbits by about 10 hours post infection and progressed in severity until by 20 h following infection, hearing losses up to and > 60 dB were obtained. At levels of hearing loss < 20 dB ultrastructural damage to the organ of Corti was barely detectable. With greater levels of hearing loss, patchy structural damage to hair cells, synaptic nerve terminals, supporting cells and inner spiral sulcus cells and cells of the stria vascularis was clearly evident. Bacteria were found in scala tympani, the basilar membrane, the organ of Corti, scala media, the spiral ligament and at the margin of the stria vascularis. Evidence of bleeding was found in some cochleas; erythrocytes were found in scala tympani, scala media, amongst hair cells and beneath the tectorial membrane. The results show that hearing loss is associated with bacterial invasion and damage to the organ of Corti and that the cause of hearing loss is likely to result from multiple lesions within the cochlea. Lesions to sensory cells almost certainly will produce permanent hearing loss. Lesions to supporting cells, nerve terminals and to stria vascularis may well produce only temporary hearing loss.

Animals↗
PubMed1994

Transcranial electrical stimulation of the motor cortex in man: further evidence for the site of activation.

1. The motor cortex was stimulated electrically (vertex anode; cathode 6 cm lateral) in neurologically normal subjects undergoing surgery for scoliosis, and the evoked corticospinal volleys were recorded from the spinal cord using epidural electrodes. 2. Stimuli > 330 V produced a complex D-wave volley containing three separate peaks, with high-threshold components, 0.8 ms (D2) and 1.6 ms (D3), in advance of the lowest-threshold component (D1). As stimuli increased up to 1500 V, D3 replaced the later components completely, but there was no further latency 'jump'. 3. Brainstem stimulation using electrodes over each mastoid process produced a descending volley that had the same latencies as D3. At threshold, stimulation of the brainstem or spinal cord attenuated the D wave evoked by simultaneous cortical stimulation. 4. It is concluded that transcranial electrical stimulation of the motor cortex at high intensities can access corticospinal neurones at the pyramidal decussation, and that stimulation of the brainstem (and the spinal cord) preferentially accesses corticospinal axons. At threshold, motor cortex stimulation probably activates corticospinal neurones at or near the cerebral cortex.

Adolescent↗
PubMed1994

Invasion of rabbit ileal tissue by Enterobacter cloacae varies with the concentration of OmpX in the outer membrane.

The outer membrane protein OmpX of Enterobacter cloacae shows high amino acid homology with virulence proteins PagC and Rck from Salmonella typhimurium and with Ail from Yersinia enterocolitica. Here we demonstrate a role for OmpX in the invasion of rabbit ileal tissue by E. cloacae. An organ culture system was used for maintenance of rabbit gut tissue during the experiments. The invasiveness of three E. cloacae strains, which differed in OmpX content, were compared with each other and with that of Salmonella typhimurium TML (a highly invasive strain) and S. typhimurium LT7 (a noninvasive strain). There was no significant difference between the invasiveness of the wild type and that of an ompX deletion mutant strain of E. cloacae; they were equally as invasive or less invasive than S. typhimurium LT7. The invasiveness of an OmpX overproducer strain of E. cloacae was 10-fold higher than that of its immediate parent carrying only the multicopy plasmid, higher than that of S. typhimurium LT7, but lower than that of S. typhimurium TML. The invasiveness of E. cloacae thus varied directly with the level of OmpX in the outer membrane in rabbit ileal enterocytes challenged in situ.

Animals↗
PubMed1994

Quantitative studies of invasion of rabbit ileal mucosa by Salmonella typhimurium strains which differ in virulence in a model of gastroenteritis.

An asymmetric organ culture system in which ileal tissues, freshly removed from rabbits, can be maintained structurally and functionally for up to 4 h has been developed. The composition of the solutions used to maintain ileal tissue in vitro were as follows. The serosal surface was bathed in the World Health Organization (WHO) rehydration formulation: NaCl, 60 mM; NaHCO3, 30 mM; KCl, 20 mM; and glucose, 111 mM. The mucosal surface was bathed in the same solution with two important changes: all the sodium was replaced by choline, which is not absorbed, and tissue culture medium (consisting of commercial minimal essential medium to which was added fetal calf serum and glutamine to final concentrations of 10% [vol/vol] and 2.0 mM, respectively) was added to the choline-containing medium to a final concentration of 10% (vol/vol). The initial invasiveness (first 2 h) of seven strains of Salmonella typhimurium differing in virulence (defined in terms of clinical origin or the ability to induce fluid loss in monkeys or rabbit ileal loops) was assessed quantitatively in an in vitro invasion assay with the organ culture system. The virulent strains (TML, W118, and WAKE) were found to be about 25- to 100-fold more invasive than the avirulent strains (SL1027, M206, LT7, and Thax-1). Thus, a clear correlation between initial mucosal invasion and virulence of S. typhimurium in a model which is relevant to human gastroenteritis was established. This is the first time, to our knowledge, that quantitative studies of invasiveness have been carried out in vitro on freshly isolated functioning gut.

Animals↗
PubMed1993

Direct comparison of corticospinal volleys in human subjects to transcranial magnetic and electrical stimulation.

1. The effects of graded transcranial magnetic and anodal electrical stimulation of the human motor cortex were compared in human subjects undergoing orthopaedic operations on the spine, before and after withdrawal of volatile anaesthesia. Corticospinal volleys were recorded from the spinal cord in the low-cervical and low-thoracic regions (six subjects) or the mid-thoracic region (two subjects) using bipolar electrodes inserted into the epidural space. 2. Electrical stimuli were delivered using anode at the vertex and cathode 7 cm laterally. The corticospinal volley at threshold consisted of a single deflection with a mean latency to peak of 4.17 ms at the rostral recording site. With further increases in stimulus strength the latency of this D wave shortened in two steps, first by 0.89 ms (seven subjects) and then by a further 0.8 ms (two subjects), indicating that the site of activation of some corticospinal neurones had shifted to deep subcortical sites. 3. When volatile anaesthetics were given, a corticospinal volley could not be defined in three subjects with magnetic stimuli of 70, 80 and 100% maximal stimulator output with the coil at the vertex (Novametrix Magstim 200, round coil, external diameter 14 cm). In the remaining five subjects, the component of lowest threshold was a D wave recorded at the rostral site at 4.0 ms when stimulus intensity was, on average, 70%. With stimuli of 90-100% a total of five small I waves could be defined in the five subjects (i.e. on average one I wave per subject). 4. After cessation of volatile anaesthetics in seven subjects, the thresholds for D and I waves were lower and their amplitudes were greater. The D wave remained the component of lowest threshold in all subjects, appearing at the low-cervical level with magnetic stimuli of 50%. However, in three subjects I waves also appeared at D wave threshold, and the D wave was smaller than with electrical stimulation at I wave threshold. There was no consistent change in latency of the magnetic D wave as stimulus intensity was increased to 100%. 5. These findings suggest that the previously reported difference in latency of the EMG potentials produced in upper-limb muscles by anodal stimulation and magnetic stimulation of the human motor cortex is not because the corticospinal volley induced by magnetic stimulation lacks a D wave.(ABSTRACT TRUNCATED AT 400 WORDS)

Adolescent↗
PubMed1993

Cytotoxic effects on hair cells of guinea pig cochlea produced by pneumolysin, the thiol activated toxin of Streptococcus pneumoniae.

The cytolytic toxin, pneumolysin, from the gram positive bacterium, Streptococcus pneumoniae, when perfused through the scala tympani of the guinea pig cochlea reduced the amplitude of both the compound action potential and the cochlear microphonic potential. When the surface of the organ of Corti was examined by scanning electron microscopy, both inner and outer hair cells and supporting cells were found to be damaged. Inner hair cells and outer hair cells of row 3 were the most susceptible to damage by pneumolysin, followed by row 2 and then by row 1 of the outer hair cells. Damage to hair cells included disruption and splaying of stereocilia, loss of stereocilia and complete dissolution of hair bundles. Apical surfaces of hair cells and supporting cells were torn, pitted and cratered with shrinkage and tearing of cell boundaries. Within the dose range perfused (0.05-1 micrograms/microliters in a 10 microliters aliquot), the magnitude of the physiological and anatomical lesions was concentration dependent. The cytotoxic effects of pneumolysin reported here may be clinically significant factors in deafness caused by meningitis and otitis media in humans.

Animals↗
PubMed1992

Moratorium-achievement (MAMA) cycles in lifespan identity development: value orientations and reasoning system correlates.

Three important questions emerging in identity research are: Why do only some persons with Foreclosed identities move on to Identity Achievement? Why do some Identity Achievement persons move "back" to Foreclosure? How can life-span identity re-formulation be accounted for? This study proposes that individual's positions on two variables, "instrumental vs. experiential orientation" and formistic vs. dialectrical thinking, may be important in answering these questions. Among 60 university students, 23 males and 37 females, those in an identity status characterized by formation and maintenance of structure (Foreclosure) were instrumentally-oriented while those characterized by openness to change and revision (Moratorium) were experientially-oriented. Similarly, more Moratoriums than other statuses were high in dialectic thinking. These results are discussed in terms of their theoretical implications for life-span identity development.

Adult↗
PubMed1992

Assessment of corticospinal and somatosensory conduction simultaneously during scoliosis surgery.

The function of descending motor pathways and that of ascending sensory pathways in the spinal cord were monitored at the same time in 120 patients undergoing surgery for scoliosis. Transcranial electrical stimulation of the motor cortex was performed simultaneously with stimulation of the tibial nerves in the popliteal fossae, and the descending and ascending volleys were recorded from the spinal cord at two levels using epidural electrodes. Stable recordings of both volleys have been obtained in all neurologically normal patients and in many with pre-existing neurological deficits. The experimental conditions which resulted in reliable recordings were explored in select patients and include: a vertex-anode/lateral cathode montage for transcranial stimulation, epidural recording of evoked corticospinal and somatosensory volleys at two spinal levels, a high-pass filter of 500 Hz, and stable anaesthesia. The epidural recording allows full muscle relaxation and the use of volatile anaesthetics; recording at two levels allows a deterioration in function to be identified quickly and distinguished from an artifactual change.

Anesthetics↗
PubMed1992

Anodal and cathodal stimulation of the upper-limb area of the human motor cortex.

In 18 neurologically normal subjects the corticofugal volleys evoked by anodal and cathodal electrical stimulation of the motor cortex were recorded using epidural electrodes at the high-thoracic and low-thoracic regions of the spinal cord during surgery for scoliosis. At and just above threshold, anodal and cathodal stimulation of the upper-limb area and motor cortex produced a D wave that propagated to the low-thoracic region. The stimulus intensity required to produce D waves was significantly lower with anodal stimulation. I waves were recorded at higher stimulus intensities than the D wave but not more readily with cathodal stimulation. There was no significant difference in the extent to which stimulus intensity had to be increased above D-wave threshold to produce I waves with the two stimulus polarities, and the number of I waves was the same when the stimulus was increased by the same amount above D-wave threshold. After withdrawal of isoflurane, I waves could not be recorded when the stimulus intensity was below D-wave threshold with either stimulus polarity. Anodal stimulation over the upper-limb area remained more effective than cathodal stimulation in producing both D and I waves. These results indicate that, at threshold, regardless of anaesthesia, anodal and cathodal stimuli access upper-limb corticospinal neurons directly at a similar site, the anodal stimuli being more effective. In addition, the results suggest that some corticospinal neurons in the upper-limb area of motor cortex have projections to lumbar segments.

Adolescent↗
PubMed1992

Corticospinal volleys evoked by electrical stimulation of human motor cortex after withdrawal of volatile anaesthetics.

1. In twenty-two neurologically normal patients undergoing surgery for scoliosis, corticospinal volleys to transcranial electrical stimulation of the motor cortex were recorded from the spinal cord using epidural electrodes. While anaesthesia was maintained by nitrous oxide and narcotics, volatile anaesthetics were withdrawn to determine whether such agents had a depressant effect on the evoked corticospinal volley. 2. Profound changes were documented in liminal D waves, there being an increase in amplitude averaging 392% following withdrawal of the volatile anaesthetic. There was a proportionately smaller increase (averaging 26%) in supraliminal D waves; these had a complex bifid or trifid shape indicating that some corticofugal axons were being activated deep to cortex. In general the effect on the D wave of withdrawing the anaesthetic agent was similar to that of increasing stimulus intensity. 3. Withdrawal of isoflurane dramatically increased the number of I waves and their mean amplitude. In the absence of isoflurane, I3 (mean latency 3.5 ms after the D wave) became the dominant I wave. The amplitude of I2 (mean latency 2.2 ms) became slightly smaller. The change in I waves could not be likened to an increase in stimulus intensity, because I waves invariably increase in, or remain of the same, amplitude as stimulus intensity is increased. 4. These findings indicate that changes in motor cortex excitability can result in major changes in the corticospinal volley produced by transcranial electrical stimulation, affecting both the D wave and I waves. They caution against identifying a cortical action solely on the basis of a change in the responses to magnetic stimulation of motor cortex but no such change to electrical stimulation.

Adolescent↗
PubMed1991

A study of the microcirculation in whole villi of neonatal mice using a peroxidase histochemical staining method.

The anatomy of the microcirculation of intestinal villi from the upper, middle, and lower small intestine of neonatal mice from 8 to 14 days old was studied using a histochemical peroxidase technique that specifically stained erythrocytes. Over 8-14 days, there was little chronological variation between the same regions of gut; the exception was the lower intestine, which, in younger mice, was noticeably less well perfused with erythrocytes. Vascular beds in the middle and lower intestine comprised a hairpin loop with cross-connections. In the upper intestine, the capillary beds were generally more complex, particularly in apical regions of the villi. Most villi were well perfused with erythrocytes, but a minority (less than 10%) contained considerably fewer red cells, some to the point of being totally ischemic. Other villi (less than 5%) were hyperemic, and the vascular beds packed and engorged with red cells. Usually, the packing density (hematocrit) of red cells within blood vessels increased progressively from villus base to apex. Red-cell deformation was more prevalent at villus apices, with marked crenation in some villi, yet in the basal regions of these same villi, the red cells were of normal discoid shape. The peroxidase staining technique produces a reliable histological picture of red cells circulating through villi. It also reveals differential perfusion of erythrocytes between and within villi, and that blood vessels pass through hypertonic zones in the apical regions of villi.

Animals↗
PubMed1991

Rotavirus-induced changes in the microcirculation of intestinal villi of neonatal mice in relation to the induction and persistence of diarrhea.

Using a histochemical peroxidase technique, under conditions that preferentially stain erythrocytes, we have shown changes in the microcirculation of villi of neonatal mice infected with murine rotavirus. Between 18 and 48 h postinfection (PI), throughout all areas of the small intestine there occurred, sequentially, a marked ischemia and atrophy of villi. By 72 h PI, villi had recovered their normal height and showed incipient hyperemic microcirculation. At 96 h PI, hyperemic microcirculation was most marked. Between 120 and 144 h PI, a second phase of villus atrophy occurred, which was more attenuated and confined to the upper and middle regions of the intestine. This phase was not accompanied by a wide-spread ischemia of villi: a minority of villi were short and ischemic but many appeared hyperemic. Recovery of villus microcirculation occurred at 168 h PI, which coincided with recovery from diarrhea. These changes in villus microcirculation are discussed in relation to the pathology and pathophysiology of rotavirus infection. We make two novel suggestions. First, the reduction in red cells flowing through villi in the early stages of the infection instigates hypoxia and hence atrophy of villi. The ensuing but ephemeral increase in rate of cell division, necessary for the reconstitution of villi, induces hypersecretion. Second, the increase in numbers of erythrocytes found in villi during their regrowth phase and throughout the remaining time course of the infection perturbs the countercurrent system, lowering the osmolality of the hyperosmotic zone located at villus tips, thereby impairing water absorption and prolonging diarrhea.

Animals↗
PubMed1991

Invasion of HEp-2 cells by strains of Salmonella typhimurium of different virulence in relation to gastroenteritis.

Experiments to measure the invasiveness of seven strains of Salmonella typhimurium for HEp-2 cells showed that high inocula (100 bacteria/HEp-2 cell), as used by most workers to synchronise events and to increase the number of bacteria which invade, resulted in recovery of significantly less than 1% of the original inoculum after treatment with gentamicin to kill extracellular bacteria. Also, the cell culture medium became acidic, and microscopic examination of Giemsa-stained monolayers immediately following gentamicin treatment revealed high concentrations of bacteria associated with the cells. Moreover, with bacterium-cell interaction beyond 2 h, many HEp-2 cells became rounded, especially with virulent strains W118 and TML. Thus, the biological significance of the quantitative data was uncertain. The fall in pH and the rounding of HEp-2 cells were prevented by the use of a low (1:1) bacterium: cell ratio; but the recovery of bacteria after treatment with gentamicin was still lower than expected by microscopic examination. After treatment of cells with Triton X-100 to release bacteria, many remained bound to residual cell nuclei. Additional treatment with a rubber policeman, and vigorous pipetting to disperse aggregates of bacteria and cell debris, increased the recovery to c. 10% of the initial inoculum after interaction for 2 h, and 30-80% after 4 h, depending on the strain and experimental conditions. The pattern of invasiveness, but not the absolute count, was highly reproducible on different days and in different hands. However, after interaction exceeding 2 h, the distribution of bacteria was uneven, many cells having no associated organisms, others showing microcolonies.(ABSTRACT TRUNCATED AT 250 WORDS)

Bacterial Adhesion↗
PubMed1990

An in vitro model to study aspects of the pathophysiology of murine rotavirus-induced diarrhoea.

An in vitro system is described and validated for studying transport of solutes and water in both uninfected and rotavirus-infected neonatal mouse intestine. Control intestine exhibited stable water absorption for periods of up to 40 min. Water absorption was temperature-dependent. Na-dependent, and enhanced by glucose-containing perfusion solutions. Theophylline induced net secretion of water by control intestinal tissue. Water transport by rotavirus-infected lower small intestine was significantly depressed as compared to control levels, and rotavirus-infected middle small intestine exhibited net secretion of water. Upper small intestine and colon from infected animals did not differ significantly from control tissues in their ability to transport water. Water secretion by infected middle small intestine was reversed to absorption by glucose-containing solutions.

Animals↗
PubMed1990

X-ray microanalysis of rotavirus-infected mouse intestine: a new concept of diarrhoeal secretion.

Neonatal mice were infected at 7 days of age with rotavirus [epizootic diarrhea of infant mice (EDIM) virus] and killed at 24-h intervals postinfection (PI). Cytoplasmic concentrations of Na, Mg, P, S, Cl, K, and Ca intestinal epithelial cells from infected and age-matched control animals were measured by x-ray microanalysis. In villus tip cells, Ca concentration increased at 24-96 h PI; Na concentration increased at 24-72 h PI; Ca and Na concentrations were near normal by 168 h PI. K concentration decreased 24-72 h PI, and Cl concentration decreased 48-96 h PI. In crypt cells, changes were observed without a discernible pattern: at 96 h PI, Na, Mg, S, and Cl concentrations increased and K concentration decreased; at 120 h PI, the concentrations of all elements except Na and Ca increased. In villus base cells, the mean concentrations of all elements except Ca peaked at 48-72 h PI and at 120 h PI. Na and Cl concentrations increased dramatically in some cells from 48 h PI onward. All the above concentration values were obtained from freeze-dried specimens and expressed in millimoles per kilogram of dry weight. Conversion of a limited number of data, pertaining to villus base cells, from dry weight to wet weight was possible. This conversion revealed that villus base cells in infected animals were more hydrated than corresponding cells from control animals. Also, the Na and Cl concentrations in mmol/kg H2O were significantly higher in villus base cells from infected animals than in those from corresponding controls: 137 +/- 7 versus 38 +/- 4 (Na) and 121 +/- 5 versus 89 +/- 6 (Cl). Wet weight concentrations of other elements were either the same (Mg) or lower (P, S, and K) after infection with virus. From these studies, a new concept of the pathophysiology of rotavirus-induced diarrhoeal secretion is proposed: stimulation of villus base cells to rapid cell division is accompanied by transient accumulation of Na and Cl; excess NaCl is secreted into the lumen, which is the driving force for fluid loss.

Animals↗