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Biomedical subjects

C J Cook

Publications and source records attributed to C J Cook.

At least 19 recordsLinked to original sources
PubMed2004

Frequencies of prion protein (PrP) genotypes and distribution of ages in 15 scrapie-affected flocks in Great Britain.

The frequencies of prion protein (PrP) genotypes were investigated in 15 scrapie-affected flocks in Great Britain. The flocks were heterogeneous in the frequencies of different genotypes and alleles, and in their age distributions. The median flock frequency of animals with VRQ-containing genotypes was 21 per cent (range 2 to 82 per cent, mean 25 per cent). The VRQ-containing and other non-ARR genotypes made up 11 to 82 per cent of a flock (median 46 per cent, mean 48 per cent). In comparison with data from the general population the scrapie-affected population had a lower frequency of the ARR/ARR genotype, and so of the ARR allele, and had a higher frequency of VRQ/non-ARR heterozygote genotypes, and thus of the VRQ allele.

Alleles↗
PubMed2004

Influence of stocking density on the performance, carcass characteristics and selected welfare indicators of broiler chickens.

AIM: To compare the effects of four stocking densities (5, 10, 15 or 20 birds per m2) on the performance, carcass characteristics and selected welfare indicators of broiler chickens. METHODS: A total of 756 day-old male broiler chicks were assigned as six replicates of 13, 25, 38 and 50 birds per pen, to stocking densities of 2,000, 1,000, 667 and 500 cm2 floor space per bird, respectively. The birds were housed in floor pens in an environmentally controlled room. Normal commercial diets were fed ad libitum and fresh water was freely available throughout the 5-week study period. Liveweights of birds and feed intake were recorded at weekly intervals. Data on carcass parameters and welfare indicators (moisture, gait scores, feather scores, breast and hip lesions, and foot pad and hock burns) were obtained on Day 35. Adrenal glands were removed at slaughter, weighed and examined histologically to determine the total proportion of cortical and medullary tissues. RESULTS: During the study, the birds at the lowest density grew faster (p<0.001-0.01) and consumed more (p<0.001- 0.01) feed than those at the other three stocking densities. The liveweight gains and feed intake of birds reared at densities of 10, 15 and 20 birds per m2 were similar (p=0.11-0.83 and p=0.17-0.57, respectively). Stocking density had no influence on the feed/weight gain, mortality or carcass characteristics. No breast or hip lesions were observed in carcasses from any of the groups. The weight and structure of the adrenal glands were similar across all groups although the staining of medullary tissue of birds was most intense in the highest stocking density group. As stocking density increased, so did moisture and gait score, hock and foot pad burn scores, and the feather score was adversely affected. CONCLUSIONS: The data showed that the relationship between stocking density and productivity traits was not linear in nature, in accordance with other studies. Under the conditions of the current study, the performance, mortality and carcass characteristics of broilers grown at densities of 10, 15 and 20 birds per m2 were similar. There were some welfare implications at high population densities as indicated by adverse effects on litter quality, gait scores and the incidence of hock and foot pad burns.

Journal Article↗
PubMed2004

Technical note: protein conjugate-based immunoassay of whole milk progesterone.

The objectives were to develop a novel protein conjugate-based ELISA test for whole milk progesterone with a dynamic range capable of fully profiling estrous cycles in the dairy cow and to study effects of whole milk medium on antibody binding to progesterone-protein conjugates. A series of progesterone-4-ovalbumin conjugates with different length intermediate linkers were applied as coating antigens in an ELISA format to determine antibody-binding performance in whole milk. Use of an 18-atom linker gave higher binding than use of a 4- or 11-atom linker at certain conjugate concentrations, but no further increase was observed with increasing linker length. An ELISA constructed with the 18-atom linker conjugate gave a detection limit of 0.089 ng/mL progesterone and correlated well to an established radioimmunoassay procedure (r = 0.94). The assay has the distinct advantages of a wide linear range (0.1 to 100 ng/mL), allowing full profiling of bovine estrous cycles, use of whole milk directly without extraction or prior dilution, and employing more easily purified protein conjugates as coating antigens compared with commercial progesterone-enzyme conjugate for milk ELISA assays.

Animals↗
PubMed2003

The LightTyper: high-throughput genotyping using fluorescent melting curve analysis.

Instrumentation, chemistry, and software for high-throughput genotyping using fluorescent melting curves are described. The LightTyper system provides post-amplification genotyping within 10 min using samples in 96- or 384-well microplate formats. The system is homogenous because all reagents are added at the beginning of the reaction and there is no sample manipulation between amplification and genotyping. High-resolution melting curves are achieved by slow and steady heating. As samples are heated, panels of blue light-emitting diodes excite the probes, and fluorescence emission is acquired with a cooled charge-coupled device camera. A variety of probe chemistries are compatible for genotyping on the LightTyper, including dsDNA dyes, single-labeled probes, and fluorescence resonance energy transfer systems. Genotyping is performed automatically, and each sample is given a score reflecting the similarity of the genotype to the standards provided. Standard genotypes can be selected from within the run or imported from other files. Samples and genotypes can be grouped to allow multiple-allele detection on one or many samples. The utility of the LightTyper is illustrated by genotyping samples for the Factor V Leiden mutation and for mutations in the CFTR gene.

Base Sequence↗
PubMed2001

Measuring of extracellular cortisol and corticotropin-releasing hormone in the amygdala using immunosensor coupled microdialysis.

A method is described for the measurement of cortisol or corticotropin-releasing hormone (CRH) from the brain, within, or in the dialysate of, a microdialysis probe using an antibody-linked assay. Polyclonal antibodies for either cortisol or CRH provide a specificity of measurement. These antibodies are affixed on a platinum electrode within the probe. Determination of bound cortisol or CRH is performed via an indirect assessment of competitive ligand also bound, and conjugated to HRP. This is activated and measured as current change. These probes were used for extracellular measurements in the amygdala, a limbic brain region, of sheep. Direct measurement in vivo compared favorably to the use of either the immunosensor or standard RIA techniques on dialysate emerging from a classical microdialysis probe, suggesting that the method could be used directly in-vivo. This is the first report in literature of rapid on-line measurement of CRH and cortisol from a discrete brain area. The probes have fast response times (>90% of maximum response within 30 s of start of analysis), high sensitivity (<0.1 pg/ml for CRH) and can acquire data every 2 min. They are stable in-vivo (>72 h) and regenerable, offering on-line measurement, with rapid time resolution, of neurohumoral substances.

Amygdala↗
PubMed1999

Patterns of weaning and adult response to stress.

Early environmental effects including variation in maternal care, can modify hypothalamic-pituitary-adrenal (HPA) axis function. One of the more overt early effects, involving maternal care, is weaning restraint. In this study the effects of different patterns of weaning, in the rat (Rattus norvegicus), on both adult response to restraint stress and to dexamethasone administration were examined. Animals that as pups experienced a gradual lengthening time of separation from the mother, between 21 and 30 days of postnatal age (completely separated on 30 days), showed lower levels of systemic corticosterone and glutamate in the sensory cortex in response to restraint stress than seen in other groups. These animals also showed greater suppression of corticosterone by dexamethasone than did animals abruptly removed from the mother at 21 days of age. Animals left in the cage with the mother until either 30 or 40 days of age showed the greatest levels of corticosterone and glutamate in the sensory cortex in response to the stress and the least suppression by dexamethasone. These results suggest that changes in maternal contact at time of weaning can influence adult responsiveness to stress.

Animals↗
PubMed1999

Maternal behaviour in sheep (Ovis aries) following administration of opioid agonists.

AIMS: As kappa(kappa)-opioids have marked effects on pain threshold in sheep during oestrus, late pregnancy and after birth, a study was undertaken to determine if kappa -opioids also had other roles during these states. METHODS: Opioid agonists (GR89696, DAMAGO) were administered into either the lateral hypothalamus (LH) or amygdala of non-pregnant sheep (n = 5) or pregnant sheep (n = 15) in late pregnancy (n = 5) within 12 h of birth (n = 5) or in the third week after birth (n = 5). Cortisol, behavioural and electrocardiographic responses to an auditory stimulus were assessed with and without drug administration, as was feed consumption and suckling events. RESULTS: In non-pregnant sheep, DAMAGO injected into either the LH or amygdala had little effect on the response to the auditory stimulus but when injected into the LH did decrease food consumption. GR 89696 had no obvious effects. In late pregnancy, cortisol and heart rate responses to the auditory stimulus were significantly reduced compared to the non-pregnant animals irrespective of treatment. This was also the case in the third week after birth. Immediately post-partum cortisol responses were closer to that seen in the non-pregnant animal. In late pregnancy and both post-partum periods, GR 89696 in the LH significantly increased feed consumption while DAMAGO had no significant effects. GR 89696 injected into the amygdala reduced the behavioural response to the auditory stimulus in late pregnancy and both post-partum periods. CONCLUSION: Changes in sensitivity to opioids occur in both the amygdala and LH with pregnancy and after birth. These may influence maternal behaviours and feed intake.

Journal Article↗
PubMed1998

Monitoring on-line of extracellular gamma-amino-4-butyric acid using microdialysis coupled to immunosensor analysis.

A method is described for the measurement of gamma-amino-4-butyric acid (GABA) within a microdialysis probe using an antibody-linked assay. A monoclonal antibody for GABA provides the specificity of measurement and these antibodies are affixed on a working platinum electrode within the probe. Determination of bound GABA is performed via an indirect assessment of competitive ligand also bound, and conjugated to horseradish peroxidase which is activated and measured as current change. Using this probe directly for extracellular measurements in the somatosensory cortex of sheep compared favorably to the use of this probe on dialysate emerging from a classical microdialysis probe suggesting that it could be used directly in vivo. The probe has a fast response time (> 90% of maximum response within 30 s of start of analysis) and high sensitivity (< 0.5 mumol/l) and can acquire data every 2 min. It is stable over a period of time in vivo (> 48 h) and is regenerable. The probe design offers on-line measurement, with rapid time resolution of substances not amenable to enzyme-based amperometric measurement.

Animals↗
PubMed1998

Influence of mouse position on muscular activity in the neck, shoulder and arm in computer users.

Anecdotal reports of shoulder and arm discomfort related to computer mouse use are common. The aim of this study of ten subjects was to examine the influence of mouse position, relative to the keyboard, on shoulder and arm muscular activity and working posture. Surface electromyography and the Rapid Upper Limb Assessment were used to determine the differences in muscle activity and posture during an editing task in each of three mouse positions. Significantly less anterior (p = 0.01) and middle (p = 0.03) deltoid electromyographic activity was demonstrated when the computer mouse was positioned adjacent to a keyboard without a numeric pad when compared to performance of an identical task with the mouse adjacent to a standard keyboard. Electromyographic activity in trapezius muscle did not differ between mouse positions. Working posture of right handed mouse users is improved by removal of the numeric keypad.

Adult↗
PubMed1998

Steroidal hormones determine sex-related differences in opioid-induced elevation of nociceptive threshold in sheep (Ovis aries).

AIMS: As sex related differences in opioid-induced elevations of nociceptive thresholds have been observed in sheep, a study was undertaken to determine if puberty, oestrus, pregnancy or administration of the hormones oestrogen and testosterone had any effect on these differences. METHODS: Withdrawal latency to a nociceptive thermal source was measured in Romney cross sheep consisting of lambs (n = 6) prior to and after their first oestrus, ram lambs (n = 6) prior to and after puberty, non-oestrous ewes (n = 6), oestrous ewes (n = 6), pregnant and subsequently suckling ewes (n = 12), wethers (n = 6) and rams (n = 6). The effects of a kappa opioid agonist (GR 89696), oestradiol benzoate, testosterone and saline on this withdrawal latency were tested in all groups. RESULTS: GR 89696 increased withdrawal latency in all animals, compared to saline, but this was most marked in ewes, particularly those in oestrus or in late pregnancy. Testosterone and oestradiol had no effect on their own, or in combination, on the withdrawal latency seen with saline alone. Oestradiol increased the effects of GR 89696 on lambs pre-pubertal ram lambs, ewes and wethers but not on post-pubertal ram lambs nor rams. Co-administration of testosterone antagonised this oestradiol effect. CONCLUSION: The efficacy of kappa opioids appears to be affected by animal sex, an effect that can be contributed to by both oestradiol and testosterone.

Journal Article↗
PubMed1997

Real-time extracellular measurement of neurotransmitters in conscious sheep.

Measurements of neurotransmitters in conscious animals have been restrictive in real-time. The use of specific enzymes within an amperometric probe, based upon a microdialysis membrane, may overcome some of these problems. I report the use of such a probe, with different perfusions of enzymes, to allow real-time measurement of glutamate, catecholamines and indoleamines, in conscious animals. At an adjacent site microdialysis sample collections were made concurrently and neurotransmitters measured in the dialysate. Both probes were positioned within the somatosensory cortex. Values obtained by amperometric probes were similar to those in dialysate samples for glutamate, catecholamines and indoleamines during basal and stimuli related collections. Amperometric measurements showed higher peak concentrations and better time resolution than dialysate sampling. This reflects sampling differences. Application of external stimuli increased extracellular concentrations of glutamate, catecholamines and indoleamines, in both forms of sampling. Dopamine measurements did not correlate well between the two forms of sampling. This may reflect the non-specificity of the enzyme (dopamine-beta-hydroxylase) used in the amperometric probe. This combination, of microdialysis and amperometry, offers a useful tool for real-time neurotransmitter studies in vivo.

Animals↗
PubMed1997

Oxytocin and prolactin suppress cortisol responses to acute stress in both lactating and non-lactating sheep.

Cortisol response to stress appears to differ between lactating and non-lactating animals. Lactating (14 d post partum) and non-lactating sheep were fitted with probes so that drugs and hormones could be infused directly into the posterior pituitary and paraventricular nucleus of the hypothalamus. The animals were also fitted with instruments to allow monitoring of heart rate, body temperature and blood cortisol levels. Their reactions to a source of acute stress (a barking dog) were then followed, with or without drug and hormone manipulation. Results in both lactating and non-lactating animals indicated shortcomings in the use of cortisol as a stress indicator. Infusing prolactin and oxytocin into either the posterior pituitary or the paraventricular nucleus of the hypothalamus suppressed cortisol responsiveness to stress in both lactating and non-lactating animals (the latter to a greater extent). In the absence of drugs, lactating animals had a slightly higher basal level of cortisol and a lower cortisol response to stress than their non-lactating counterparts. Despite suppression of cortisol responses, with or without drugs, other indicators of stress still changed with the presence of a barking dog, suggesting the complexity of control involved in stress responses.

Animals↗
PubMed1997

Real-time measurements of corticosteroids in conscious animals using an antibody-based electrode.

An electrochemical immunosensor for real-time determination of corticosteroids, a group of steroidal hormones, is reported. The sensor measures competitive binding of endogenous corticosteroid and a corticosteroid-peroxidase conjugate with antibodies, immobilized on a platinum electrode, by monitoring of peroxidase activity. The electrode is encased within a dialysate membrane, which separates the electrode environment from the sampled fluid, allowing corticosteroid to equilibrate across the membrane. This permits measurements to be made in vivo. The small size of the probe (350 microns D) allows implantation into tissue or circulatory systems. The electrode was viable for 200 to 400 sequential measurements and at least 48 h in vivo. Detection sensitivity is 0.2 to 0.6 microgram/100 ml of cortisol or corticosterone. The results suggest the possibility of monitoring hormones in real-time within the blood or organ systems of conscious animals.

Animals↗
PubMed1996

Social behaviour in sheep relates to behaviour and neurotransmitter responses to nociceptive stimuli.

Sheep in the field display differences in social behaviour. These differences allow a division into three social groups with distinct behavioural occurrences and frequencies. The behavioural and neurotransmission responses of each of these groups to aversive stimuli were compared. Behavioural responses were seen to both forelimb electric shocks and thermal heating of the nose in all groups. These responses changed with stimulus repetition in a group-dependent manner. Microdialysis probe studies of neurotransmitter release in the somatosensory cortex indicated neurotransmitter responses to stimuli in all animals that varied with both animal group and stimulus repetition. Group 1 animals, aggressive and socially active, showed increases in gamma amino-4-butyric acid (GABA) with initial stimulus presentation; this increased with stimulus repetition. Behavioural responses to the stimuli decreased with repetition and nonstimulus-related behaviours, during the course of the experiment, increased. Both of these appeared dependent upon GABA. Group 2 animals, moderately aggressive and socially active, released opioid-like peptides (OLP) upon initial exposure to stimuli but, with repetition, switched to using GABA. Group 3 animals, nonaggressive and socially inactive, released OLP with initial and repeat stimuli. In groups 2 and 3, both GABA and OLP appear to reduce stimulus-related behaviour, but OLP appeared to also reduce nonstimulus-related behaviour and GABA increased these. Changes were independent of animal liveweight. Glutamate was released in response to stimuli in all 3 groups and, with repetition, fell in groups 1 and 2 but increased in group 3. An animal's social behaviour and status may predict its response to a stimulus.

Animals↗
PubMed1996

Changes in the release of amino acid neurotransmitters in the brains of calves and sheep after head-only electrical stunning and throat cutting.

In calves aged two to five months, throat cutting resulted in an increase in the concentration of the amino acid neurotransmitters glutamate and aspartate in the brain. Electrical head-only stunning by itself also increased the concentrations of these two neurotransmitters. The levels induced by stunning resulted in a seizure state characterised by epileptiform-like activity in the electroencephalograph. Combing head-only stunning with throat cutting within 10 seconds of the stun had a synergistic effect upon glutamate and aspartate, increasing their concentration by a greater amount and more quickly than either procedure on its own. An irreversible loss of brain function also occurred more quickly than after throat cutting alone. The administration of glutamate and aspartate receptor antagonists before the throat cutting lengthened the time to the loss of brain function in a dose dependent manner. Similar changes were observed in sheep but they occurred much more quickly than in cattle.

Abattoirs↗