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Biomedical subjects

D Jordan

Publications and source records attributed to D Jordan.

At least 19 recordsLinked to original sources

Relationships between the density of different indicator organisms on sheep and beef carcasses and in frozen beef and sheep meat.

AIM: To describe the relationship between the concentration of different indicator bacteria in red meat. METHODS AND RESULTS: Enumeration data for aerobic plate count (APC), Enterobacteriaceae, coliforms and Escherichia coli biotype I were analysed from an Australia-wide survey of beef carcasses, sheep carcasses, frozen beef and frozen sheep meat. In all commodities, there was only low-to-moderate rank correlation (0.16-0.47) between concentration of APC and concentration of each Gram-negative indicator. Rank correlations between counts of Gram-negative indicators were much higher (0.47-0.92) especially when nondetections were excluded from analysis (0.78-0.94). Receiver-operator characteristics analysis showed that detection of coliforms can predict the presence of E. coli biotype I with almost 100% sensitivity but fails to predict absence in 2.7-8.5% of samples not containing E. coli biotype I. CONCLUSIONS: Enumeration of coliforms is a useful adjunct to enumeration of E. coli biotype I or Enterobacteriaceae in red meat. The density of coliforms or Enterobacteriaceae can be used to predict the presence or absence of E. coli biotype I, although when the latter is at low prevalence errors in positive test prediction can be large. SIGNIFICANCE AND IMPACT OF THE STUDY: A quantitative basis is provided for comparing the concentration of different indicator bacteria measured in the production, regulation and trade of red meat.

Animals↗

Mass screening for antimicrobial resistant Escherichia coli in dairy cows in northern New South Wales.

OBJECTIVE: To describe aspects of the epidemiology of antimicrobial resistance in Escherichia coli shed in the faeces of milking cows in a dairying region of New South Wales. DESIGN: A survey based on multi-stage sampling with repeated measures made within herds for estimating within-herd correlation of resistance status, and with repeated measures made on identical specimens for estimating test-retest reliability. PROCEDURE: From a population of 110 dairy herds, 30 were selected at random and from each herd between 5 and 10 faecal specimens were obtained from fresh manure pats. E coli from faecal specimens were grown on hydrophobic grid membrane filters (HGMF) and replicated onto chromogenic agar and agar containing antimicrobials (gentamicin, ampicillin, tetracycline and sulfamethoxazole). Image analysis was used to assess colony growth. Data were analysed descriptively, by generalised linear mixed models and by Taylor series linearisation to account for attributes of the survey design. RESULTS: Of the 10,279 E coli isolates assessed, 91% expressed no resistance, 7.3% were resistant to sulfamethoxazole, 3.6% to tetracycline, 2.2% to ampicillin and 0.09% to gentamicin. The most common multiple resistance phenotype was ampicillin-tetracycline-sulfamethoxazole (1.8% of isolates). Most multiple resistant isolates appeared clustered within particular herds but were too rare to obtain valid estimates of variance, confidence intervals or intra-herd correlation. The estimated proportion of isolates in the population that were susceptible to all four antimicrobials was 97% (95% CI: 91% to 100%) and 55% of cows had no resistance detected in faecal E coli (95% CI: 27% to 83%). Within-herd correlation of shedding status (any resistance pattern) was absent and test-retest reliability of the measurement system was estimated to be at the lower end of good (0.40) but increased to excellent (0.89) after excluding sulfamethoxazole resistance, which had a greater measurement error. CONCLUSION: Antimicrobial resistance was uncommon in E coli in the population of dairy cows studied. HGMF and image analysis is an effective tool for detecting rare forms of resistant E coli that are not uniformly distributed in livestock populations.

Animals↗

The relationship between concentration of a dual marker strain of Salmonella Typhimurium in bovine faeces and its probability of detection by immunomagnetic separation and culture.

AIMS: To modify a strain of Salmonella serotype Typhimurium to express unique marker traits and then define how the concentration of the marker in bovine faeces affects the probability of its detection by culture preceded by immunomagnetic separation (IMS). METHODS AND RESULTS: DNA encoding for the production of green fluorescent protein (gfp) and resistance to kanamycin was inserted into the bacterial chromosome of Salm. Typhimurium. Transposon insertion was demonstrated by Southern blot hybridization. Varying amounts of one electroporant (gfpSal-1) were inoculated into suspensions of bovine faeces and attempts made to isolate gfpSal-1 using a protocol based on pre-enrichment incubation, IMS and enrichment in selective media. Isolates of gfpSal-1 were differentiated from wild strains of Salmonella using fluorescence under u.v. light and expression of kanamycin resistance. A logistic and Gompertz function each derived from the dose-response data partially explained the observations with the fit of the Gompertz function judged to be superior. The 10, 50 and 90% limits of detection from the Gompertz function were estimated to be 1.92, 2.03 and 2.27 CFU g(-1) respectively. CONCLUSIONS: Reliance on the traditional concept of 'limit of detection' could introduce unacceptable errors in the interpretation of test findings when the concentration of Salm. Typhimurium in bovine faeces (pooled or individual) is below ca 3 CFU g(-1) of faeces. SIGNIFICANCE AND IMPACT OF THE STUDY: The dose-response curve can be used to aid the design of protocols for detecting Salmonella in individual and pooled faecal specimens. The experiments demonstrate that both reporter genes in tandem are useful for studying the performance of culture-based methods for detecting pathogens in faeces.

Animals↗

Physiological, pharmacological, and immunohistochemical characterisation of juxtacellularly labelled neurones in rat nucleus tractus solitarius.

The pharmacology and anatomy of neurones in the nucleus tractus solitarius (NTS) have proved to be difficult to study in vivo because of their generally small size and high packing density. To overcome these problems, we have developed an approach that combines drug application through multibarrelled electrodes with juxtacellular labelling via an attached single-barrelled electrode followed by immunohistochemical processing. This approach has allowed us to assess the responses of individual NTS neurones in vivo to ionotropic glutamate receptor agonists and antagonists and then, to determine whether the neurones expressed the glutamate receptor subunits, GLUR2,3 and NMDAR2a,b. It should also be possible to extend these techniques further and correlate morphology with these features and to examine pharmacologically characterised, dye-filled neurones at the ultrastructural level.

Animals↗

Effect of stimulating non-myelinated vagal axons on atrio-ventricular conduction and left ventricular function in anaesthetized rabbits.

It has previously been demonstrated in several species that stimulation of myelinated vagal efferent fibres evokes slowing of heart rate and atrio-ventricular (A-V) conduction and a decreased ventricular contractility but recruitment of non-myelinated fibres did not further increase the response. Only in rabbits was a significant bradycardia evoked on recruiting non-myelinated fibres. However, if stimulating myelinated fibres produced a near maximal response, then effects of further activation of non-myelinated fibres may have been missed. Indeed, selective stimulation of non-myelinated fibres alone now has been shown to evoke a slowing of heart rate independent of the effects of myelinated fibres. In the present study we tested in rabbits whether selective stimuli are also capable of slowing A-V conduction and changing ventricular contractility. In rabbits pretreated with the beta 1-adrenoceptor antagonist atenolol, ECG, arterial blood pressure, left ventricular pressure and dP/dt were recorded before and during stimulation of non-myelinated vagal efferent fibres using an anodal block technique (J. Physiol. 273 (1977) 539). R-R interval and A-V conduction times were computed off-line. Stimulation of non-myelinated vagal fibres increased R-R interval by 97.7 +/- 18.8 ms from a baseline of 315.3 +/- 7.7 ms, increased A-V conduction time by 9.9 +/- 1.1 ms from a baseline of 81.9 +/- 3.1 ms and decreased left ventricular dP/dtmax by 2486 +/- 362 mmHg s-1 from a baseline of 11,186 +/- 795 mmHg s-1. When hearts were paced at a rate about 10% higher than normal, A-V conduction time still increased by 13.3 +/- 1.9 ms from a baseline of 104.2 +/- 3.6 ms and dP/dtmax still fell by 2300 +/- 188 mmHg s-1 from a baseline of 11,200 +/- 777 mmHg s-1. Ganglionic blockade with hexamethonium (15-20 mg kg-1) always abolished the evoked increases in A-V conduction time, whilst there was still an increase in R-R interval in seven of the 12 animals tested. The data demonstrate that non-myelinated vagal efferent fibres can modulate chronotropic, dromotropic and inotropic actions on the heart.

Anesthetics↗

Central nervous pathways and control of the airways.

Neural control of airway muscles and secretions is predominantly by excitatory parasympathetic and non-adrenergic, non-cholinergic innervations (excitatory and/or inhibitory depending on the species). Functionally distinct afferents effecting airway reflexes terminate in different but overlapping parts of the nucleus tractus solitarius, where integration of simultaneously evoked reflex responses occurs. Parasympathetic preganglionic neurones are located in the dorsal vagal nucleus and nucleus ambiguus, which also contains upper airway motoneurones. These output neurones receive inputs from the central respiratory network which modify the effectiveness of reflex activity. This is particularly important since many afferents evoking airway reflexes concurrently modify respiratory drive. Thus, their effect on the outflow is twofold, a direct reflex effect and an indirect respiratory action and these may facilitate or antagonise one another. Although there is reflex control of individual motor outflows, in some defined situations, e.g. swallowing and coughing a stereotypical pattern of motor outflow is evoked. The neural mechanisms underlying these aspects of airway control are discussed.

Animals↗

Distal forearm fracture history in an older community-dwelling population: the Nottingham Community Osteoporosis (NOCOS) study.

OBJECTIVES: to assess the prevalence of a history of Colles' fracture (occurring after the age of 40 years) and to ascertain the extent of investigation and treatment of osteoporosis in this population. METHODS: we studied subjects aged > or =60 years from the age-sex register of three general practices. We recorded a history of fractures and details of any previous investigation for osteoporosis and treatment with bone-protective drugs. Bone mineral density was performed at the heel using dual-energy x-ray absorptiometry (Lunar PIXI machine). We classified subjects into normal, osteopaenic or osteoporotic according to the machine manufacturer's recommended World Health Organisation 'equivalent T-score thresholds' (0.6 for osteopaenia and 1.6 for osteoporosis). RESULTS: of the 605 subjects invited, we recruited 259 women and 194 men (response rate=74.8%). Twenty-eight (10.8%) of the women and five (2.6%) of the men had a history of Colles' fracture. Of women with a prevalent Colles' fracture, 39% were osteoporotic and 36% were osteopaenic. These rates were significantly greater than in women without a Colles' fracture (19.9% osteoporotic, 29.4% osteopaenic; P=0.018). Assuming the same PIXI thresholds for men, two (40%) of the five men with a history of Colles' fractures were osteoporotic and the rest were osteopaenic, compared with 20.6 and 31.2% of men without a history of Colles' fractures. None of the subjects in the Colles' fracture group had previously been investigated with bone densitometry. Women with and without a history of Colles' fracture did not differ significantly in ever having (32.1% vs 27.2%; P=0.4) or currently having (14.3% vs 10.4%; P=0.4) hormone replacement treatment. None of the men and only one woman with a previous Colles' fracture had ever taken a non-hormone replacement treatment for osteoporosis. CONCLUSIONS: older community-dwelling subjects with previous Colles' fracture have a high prevalence of osteoporosis and are under-investigated and under-treated. Methods for identifying subjects with a previous Colles' fracture need to be developed in primary and secondary care.

Aged↗

In vivo modulation of nucleus tractus solitarius (NTS) neurones by activation of 5-hydroxytryptamine(2) receptors in rats.

In in vivo experiments, 5-hydroxytryptamine (5-HT) and (+/-)-2,5-dimethoxy-4-iodoamphetamine HCl (DOI), a 5-HT(2) receptor agonist, were applied by ionophoresis to rat nucleus tractus solitarius (NTS) neurones identified by their vagal and cardiopulmonary afferent inputs to test whether the response of NTS cells to 5-HT(2) receptor activation was related to whether they received mono- or polysynaptic vagal inputs and their presumed function as defined by their afferent input. Cells were classified on the basis of the variability of the latency of the vagal-evoked spikes: this varied by less than 3 ms for Group 1, from 3 to 5 ms for Group 2, and more than 5 ms for Group 3. Both 5-HT and DOI inhibited most Group 1 cells (16/18) and inactive (without ongoing activity) cells (8/13) in Group 2. Cells inhibited by DOI were also inhibited by cardiopulmonary afferent stimulation, evoked by atrial phenylbiguanide administration. By contrast, application of 5-HT and DOI excited the majority of Group 3 cells (14/19) and Group 2 with ongoing activity (7/9). Cells excited by DOI were also activated by cardiopulmonary stimulation. Both actions of DOI were reversed by application of ketanserin (n=15). In conclusion, these data demonstrate that activation of 5-HT(2) receptors in the NTS produces different effects dependent on whether the neurones received mono- or polysynaptic vagal input and their response to cardiopulmonary afferent stimulation.

Afferent Pathways↗

Effect of pulmonary C-fibre afferent stimulation on cardiac vagal neurones in the nucleus ambiguus in anaesthetized cats.

It has been demonstrated previously that the vagal bradycardia evoked by activation of pulmonary C-fibres is not respiratory modulated. Experiments were carried out in alpha-chloralose anaesthetized cats to determine if these cardiac vagal preganglionic neurones (CVPNs) in the nucleus ambiguus (NA), which have respiratory modulated activity, can be activated when pulmonary C-fibre afferents are stimulated by right atrial injections of phenylbiguanide (PBG). Eleven CVPNs with B-fibre axons in the right cardiac vagal branches were identified and found to be localized within or ventrolateral to the nucleus ambiguus. Ionophoretic application of a high current of dl-homocysteic acid (DLH) induced a vagally mediated bradycardia and hypotension in six of eight sites from which CVPNs were recorded. The activity of B-fibre CVPNs, whether spontaneous (n = 4) or induced by ionophoresis of DLH (n = 7) was respiratory modulated, firing perferentially during post-inspiration and stage 2 expiration. This activity also correlated with the rising phase of the arterial blood pressure wave consistent with these CVPNs receiving an arterial baroreceptor input. Right atrial injections of PBG excited nine of eleven CVPNs tested. In eight of these activated neurones the onset latency of the excitation was within the pulmonary circulation time, consistent with being activated only by pulmonary C-fibre afferents. In two neurones the PBG-evoked excitation still occurred when central inspiratory drive was inhibited, as indicated by the disappearance of phrenic nerve activity. In conclusion, B-fibre respiratory modulated CVPNs can be activated following stimulation of pulmonary C-fibre afferents.

Action Potentials↗

Inhibition of rat nucleus tractus solitarius neurones by activation of 5-HT2C receptors.

In vivo, nucleus tractus solitarius (NTS) neurones receiving monosynaptic vagal input and inactive intermediate neurones were inhibited by both DOI and a selective 5-HT2C receptor agonist, MK-212. Cells receiving a more polysynaptic input were excited by DOI and although MK-212 also excited a few of these cells, the majority of cells in these groups were unaffected by MK-212. The inhibitory, but not the excitatory actions of both MK-212 and DOI were prevented by a selective 5-HT2C receptor antagonist, RS-102221. In contrast, most dorsal vagal preganglionic neurones were unaffected by application of either DOI or MK-212, the few remaining cells being excited by both agonists. These data demonstrate that DOI-evoked inhibition of NTS cells activated by vagal afferent input and DOI-evoked excitation of vagal preganglionic neurones is mediated by 5-HT2C receptors.

Amphetamines↗

The first case of new variant Creutzfeldt-Jakob disease in France: clinical data and neuropathological findings.

Clinical data and autopsy findings in a case of new variant Creutzfeldt-Jakob disease (vCJD) are reported. This case, the first histologically confirmed case described outside the United Kingdom, very much resembles the cases described by Will et al. [(1996) Lancet 347:921-925] and Zeidler et al. [(1997) Lancet 350:903-908, 908-910]. Neuropathological studies failed to reveal any conspicuous clues that could be relevant for understanding the pathophysiology of the disease. For epidemiological surveillance, neuropathologists should scrutinize suspected cases keeping in mind the possibility of vCJD.

Adult↗

In vivo modulation of vagal-identified dorsal medullary neurones by activation of different 5-Hydroxytryptamine(2) receptors in rats.

1. In in vivo experiments, DOI (a 5-HT(2) receptor agonist), MK-212 (a 5-HT(2C) receptor agonist), and BW-723C86 (a 5-HT(2B) receptor agonist) were applied by ionophoresis to neurones in the rat nucleus tractus solitarius (NTS) receiving vagal afferent input. 2. The majority of the putative 'monosynaptically' vagal activated cells were inhibited by both MK-212 (4/6) and DOI (2/4), but unaffected by BW-723C86 (12/14). In contrast, 'polysynaptically' activated NTS cells were excited by both BW-723C86 (13/19) and DOI (9/10). Inactive 'intermediate' cells were inhibited by BW-723C86 (9/12), MK-212 (5/6) and DOI (3/4), whilst active cells of this group were excited by BW-723C86 (7/13) and DOI (5/5). 3. The selective 5-HT(2B) receptor antagonist LY-202715 significantly reduced the excitatory actions of BW-723C86 on 'intermediate' and 'polysynaptic' cells (13/13), but not the inhibitory effects observed on inactive Group 2 cells (n=5) whereas the selective 5-HT(2C) receptor antagonist RS-102221 reversed the inhibitory effects of MK-212 and DOI on 'monosynaptic and 'intermediate' neurones. 4. Cardio-pulmonary afferent stimulation inhibited two of four putative 'monosynaptically' activated calls and all four inactive intermediate cells. These were also inhibited by DOI and MK-212. In contrast, cardio-pulmonary afferents excited all five active intermediate cells and all six putative 'polysynaptically' activated NTS cells, while all were also previously excited by BW-723C86 and/or DOI. 5. In conclusion, these data demonstrate that neurones in the NTS are affected differently by 5-HT(2) receptor ligands, in regard of their vagal postsynaptic location, the type of cardio-pulmonary afferent they receive and the different 5-HT(2) receptors activated.

Afferent Pathways↗

Structure and expression of the variant melanin-concentrating hormone genes: only PMCHL1 is transcribed in the developing human brain and encodes a putative protein.

PMCHL1 and PMCHL2 are two copies of the so-called variant melanin-concentrating hormone (MCH) gene that are located, respectively, on human chromosome 5p14 and 5q13 and that emerged recently during primate evolution. They correspond to a 5'-end truncated version of the MCH gene mapped on chromosome 12q23 and encoding a neuropeptide precursor. The gene organization and regulation of the expression of the variant MCH genes in the human brain are the central issues we investigated. First, the structure and fine chromosomal mapping of the 5p and 5q variant MCH genes were established. These revealed several point mutations and length variations of one CA/TA repeat which allow discrimination between each copy. Using a combination of RACE-PCR, RT-PCR, and sequencing analysis, we provided strong evidence for the expression of the PMCHL1 gene but not the PMCHL2 gene in the human fetal, newborn, and adult brains. Sense, potentially coding, RNAs, as well as noncoding antisense RNAs, were identified and displayed a region-specific expression in the human brain. Strikingly, sense unspliced RNAs of the PMCHL1 gene carried a novel open reading frame and may produce an NLS-containing protein of 8 kDa named VMCH-p8. These transcripts were translated in vitro and in transfected COS cells. Therefore, the PMCHL1 gene provides a unique example of the generation of a gene in the Hominoidae lineage which is specifically transcribed in the developing human brain and has the capacity to be translated into a putative novel protein.

Amino Acid Sequence↗

Leaky scanning is the predominant mechanism for translation of human papillomavirus type 16 E7 oncoprotein from E6/E7 bicistronic mRNA.

Human papillomaviruses (HPV) are unique in that they generate mRNAs that apparently can express multiple proteins from tandemly arranged open reading frames. The mechanisms by which this is achieved are uncertain and are at odds with the basic predictions of the scanning model for translation initiation. We investigated the unorthodox mechanism by which the E6 and E7 oncoproteins from human papillomavirus type 16 (HPV-16) can be translated from a single, bicistronic mRNA. The short E6 5' untranslated region (UTR) was shown to promote translation as efficiently as a UTR from Xenopus beta-globin. Insertion of a secondary structural element into the UTR inhibited both E6 and E7 expression, suggesting that E7 expression depends on ribosomal scanning from the 5' end of the mRNA. E7 translation was found to be cap dependent, but E6 was more dependent on capping and eIF4F activity than E7. Insertion of secondary structural elements at various points in the region upstream of E7 profoundly inhibited translation, indicating that scanning was probably continuous. Insertion of the E6 region between Renilla and firefly luciferase genes revealed little or no internal ribosomal entry site activity. However when E6 was located at the 5' end of the mRNA, it permitted over 100-fold-higher levels of downstream cistron translation than did the Renilla open reading frame. Internal AUGs in the E6 region with strong or intermediate Kozak sequence contexts were unable to inhibit E7 translation, but initiation at the E7 AUG was efficient and accurate. These data support a model in which E7 translation is facilitated by an extreme degree of leaky scanning, requiring the negotiation of 13 upstream AUGs. Ribosomal initiation complexes which fail to initiate at the E6 start codon can scan through to the E7 AUG without initiating translation, but competence to initiate is achieved once the E7 AUG is reached. These findings suggest that the E6 region of HPV-16 comprises features that sponsor both translation of the E6 protein and enhancement of translation at a downstream site.

5' Untranslated Regions↗

Routine outcome monitoring in a public mental health system: the impact of patients who leave care.

OBJECTIVE: An interest exists in using patient outcome data to evaluate the performance of publicly financed mental health organizations. Because patients leave these organizations at a high rate, the impact of patient attrition on routinely collected outcome data was examined. METHODS: In one county mental health system, routinely collected data on a wide range of outcomes were examined, and a random sample of patients who left treatment was interviewed. RESULTS: Of the 1,769 patients in ongoing treatment during a one-year period, 554 (31 percent) were lost to follow-up. Among a random sample of 102 patients who left treatment, two had died and 47 were interviewed. Compared with patients who left treatment, patients who stayed were older, more likely to have schizophrenia, less likely to be married, more likely to be living in an institution, more satisfied with their relationships with friends and family, and less likely to have legal problems. Average outcomes improved both for patients who stayed and for patients who left. Patients who left and could be located for follow-up were less severely ill and showed the greatest improvement and the best outcomes. Patients who left and could not be located may have been more severely ill at baseline. CONCLUSIONS: Outcomes appear to vary substantially by whether patients stay in care and whether they can be located after leaving care. Public mental health systems that wish to evaluate treatment quality using outcome data should attend carefully to which patients are being assessed. Biases can result from convenience sampling and from patients leaving care.

Adult↗