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Biomedical subjects

D Clark

Publications and source records attributed to D Clark.

At least 217 records · Page 12Linked to original sources
PubMed1992

Differences in energy metabolism between normal weight 'large-eating' and 'small-eating' women.

Nine 'large-eating' (approximately 12 MJ/d) and nine 'small-eating' (approximately 5.3 MJ/d) women were selected from the population on the basis of diet and activity diaries. At rest and in the post-absorptive state the rate of oxygen consumption (VO2)/kg fat-free mass (FFM) and rate of carbon dioxide production (VCO2)/kg FFM were 9-17% higher (P < 0.05) in the 'large-eaters' than in the 'small-eaters'. As energy expenditure was increased by walking at 2.4, 3.9 and 5.4 km/h the differences between the two experimental groups for both VO2/kg FFM and VCO2/kg FFM were decreased to negligible values, but energy expended on a body-weight basis (MJ/kg per min) remained significantly higher (5-10%) in 'large-eaters'. Oral temperature was also consistently higher (up to 0.5 degrees) in this group both at rest and during sitting, standing and walking activities. Although the average thermic effect of a standardized liquid meal tended to be higher (27%; not significant) in the 'small-eaters', the other results demonstrate that the 'large-eating' females had a markedly higher rate of energy expenditure at rest and during light physical activities.

Adult↗
PubMed1992

Monoclonal antibody to Pneumocystis carinii. Comparison with silver stain in bronchial lavage specimens.

Monoclonal 3F6 anti-Pneumocystis carinii antibody (MAB-3F6) was used to stain cell blocks from 164 bronchial lavage specimens from patients with the acquired immune deficiency syndrome (AIDS) and AIDS-related complex and compared with slides stained with Grocott's modification of the Gomori methenamine silver stain. Pneumocystis organisms were present in 83 of 164 cases using MAB-3F6 stain, whereas Grocott's modified silver stain demonstrated Pneumocystis organisms in 48. MAB-3F6 demonstrated Pneumocystis organisms in 38 cases with negative silver stains, whereas silver stain identified Pneumocystis organisms in only three MAB-3F6-negative cases. Of 70 patients with clinical Pneumocystis pneumonia at the time of the specimen was obtained, 59 had MAB-3F6-positive specimens, whereas 39 had organisms detected using Grocott's modified silver stain. Of 37 patients without clinically apparent Pneumocystis pneumonia any time in their course, 4 had abundant organisms and 33 had negative stains with MAB-3F6. MAB-3F6 detected Pneumocystis organisms in 22 of 31 cases of Pneumocystis pneumonia that had no organisms identified using Grocott's silver stain (X2 = 5.76, P = 0.016). MAB-3F6 immunochemical staining is a more sensitive method than Grocott's modified silver stain to detect Pneumocystis organisms.

Antibodies, Monoclonal↗
PubMed1992

Developmental regulation of globin gene expression.

We have used the globin family of genes in chicken to study developmental regulation of gene expression, both at the level of individual interaction of trans-acting factors with local promoters and enhancers, and at the level of chromatin structure. Regulation of all members of the alpha- and beta-globin clusters is affected by the erythroid regulatory factor GATA-1. Separate mechanisms exist for regulation of individual members of the family. As an example, we describe the control mechanisms that play a role in the expression of the rho-globin gene, which is expressed only in primitive lineage erythroid cells. In addressing the involvement of chromatin structure in gene activation, we have examined the role of locus control elements, and also considered the way in which RNA polymerase molecules might accommodate to the presence of nucleosomes on transcribed genes.

Animals↗
PubMed1992

Agonist and antagonist activity of low efficacy D2 dopamine receptor agonists in rats discriminating d-amphetamine from saline.

The ability of the low efficacy D2 agonists preclamol and SDZ 208-911 to both antagonise, and substitute for, the d-amphetamine discriminative cue was investigated in rats trained to discriminate d-amphetamine (0.5mg/kg) from saline. All doses of preclamol (2.0-16.0mg/kg) and SDZ 208-911 (0.125-1.0mg/kg) only partially antagonised d-amphetamine discrimination. In contrast, the lower efficacy D2 agonist SDZ 208-912 completely blocked the cueing properties of d-amphetamine. Preclamol (4.0 and 16.0mg/kg) and SDZ 208-911 (1.0mg/kg) also partially substituted for d-amphetamine. In an additional study, the former drug enhanced the discriminative effects of a low dose of d-amphetamine (0.125mg/kg), whilst antagonising the effects of the training dose. Preclamol also partially antagonised the ability of the selective D2 agonist quinpirole (0.125mg/kg) to substitute for d-amphetamine. In contrast to the drug discrimination findings, preclamol completely antagonised the locomotor hyperactivity induced by acute d-amphetamine, in animals which had received the same long-term d-amphetamine treatment as the drug discrimination rats. The present findings reveal that preclamol and SDZ 208-911 can exert both agonist and antagonist activity in animals trained to discriminate d-amphetamine from saline. This partial agonist profile is probably due to the low efficacy D2 agonists interacting with a postsynaptic D2 receptor population possessing a higher response capability than those D2 receptors mediating d-amphetamine-induced locomotor hyperactivity.

Journal Article↗
PubMed1992

In vitro fertilisation in immunological infertility.

Fertilisation occurs through a complex and orderly sequence of changes in both the oocyte and the spermatozoon. Although mechanisms have evolved to protect spermatozoa from recognition by the immune system, there is evidence that a breakdown in protection can occur resulting in homologous or heterologous antisperm antibodies which can immobilise or agglutinate spermatozoa leading to infertility. Antisperm antibodies may affect the reproductive process by interfering with sperm survival and motility and sperm transport through cervical mucus. Fertilisation is also affected, and those oocytes that do fertilise go on to develop poorly. Antisperm antibodies appear to inhibit sperm attachment to and penetration of the zona pellucida. Immune recognition of sperm antigens, which are normally present on the surface of the embryo once fertilisation has occurred, may lead to abortion after implantation. Despite the adverse effects of antisperm antibodies on fertilisation and embryo development, acceptable pregnancy rates can be achieved with assisted reproductive techniques using sperm-washing to remove antibody and using a protein source other than female serum. Further study is necessary to understand the mechanism of immunological infertility and to evaluate newer techniques using micromanipulation of gametes to achieve fertilisation.

Antibodies↗
PubMed1992

Results of the multicenter evaluation of the CEDIA cortisol assay.

The present paper describes the multicenter evaluation of the CEDIA Cortisol test for total cortisol. The observed linearity of the test was between 1.2 and 50 micrograms/dL cortisol. The limit of detection was calculated as 1.2/dL. Imprecision studies covering the diagnostically relevant range (5-20 micrograms/dL cortisol) yielded coefficients of variation between 1.7-8.9% (within-run) and 2.7-10.5% (between-day). An interlaboratory survey using 41 human samples and three control sera demonstrated that the new CEDIA Cortisol assay has a good interlaboratory transferability. Method comparison studies between the CEDIA Cortisol test and EIA, FIA, FPIA, and various RIAs yielded an acceptable level of agreement and concordant results in most cases. Low cross-reactivity of the antibody used in the new cortisol assay was observed with precursors or metabolites of cortisol. Especially, dexamethasone did not cross-react. However, prednisolone, 6-methylprednisone, and corticosterone showed cross-reactivities. No limitation by endogenous interferences was observed. The CEDIA Cortisol assay permits the precise, fast and sufficiently specific determination of cortisol. Furthermore, it offers the advantages of a non-radioactive assay and can be performed conveniently on Boehringer Mannheim/Hitachi analyzers in combination with routine clinical chemistry.

Humans↗
PubMed1992

Evaluation of CNBr, FMP and hydrazide resins for immunoaffinity purification of factor IX.

The American Red Cross has developed an immunoaffinity chromatography method to purify human coagulation Factor IX to high levels of purity for therapeutic treatment of hemophilia B. The resin currently used in this process is Sepharose CL2B, a cross-linked 2% agarose, which is activated with cyanogen bromide to immobilize an anti-Factor IX monoclonal antibody. This study evaluated two alternative resins and coupling chemistries, a synthetic polymer bead activated by 2-fluoro-1-methyl-pyridinium toluene 4-sulfonate (FMP) and a cross-linked 2% agarose bead with free hydrazide groups for site-specific coupling. The cyanogen bromide and FMP chemistries immobilize the monoclonal antibody in a random orientation. In hydrazide coupling, the monoclonal antibody is immobilized by the non-antigen-binding part of the molecule which, theoretically, should increase the amount of immobilized monoclonal antibody able to bind antigen. To examine this, the capacity of the resins to bind Factor IX and the purity and recovery of Factor IX eluted from the resins were measured. The FMP-activated resin exhibited the lowest capacity, binding only 2% of the Factor IX feed. Sepharose CL2B bound 87% of the loaded protein, while the hydrazide resin bound 43%. These results suggest that (a) hydrazide activation may be insufficient to orient monoclonal antibody and (b) other factors such as steric hindrances and diffusional resistances during immobilization may be important. Neither of the other resins tested demonstrated improved performance compared with cyanogen bromide-activated Sepharose CL2B for the immunoaffinity purification of Factor IX.

Antibodies, Monoclonal↗
PubMed1991

Role of dopamine D1 and D2 receptors in mediating the d-amphetamine discriminative cue.

The role of D1 and D2 dopamine (DA) receptors in mediating the discriminative cue produced by d-amphetamine (0.5 mg/kg) in rats has been assessed by using compounds which exert strong selectivity for each of these DA receptor subtypes. The D2 agonists quinpirole and RU 24213 substituted completely for d-amphetamine, while the D1 agonists SKF 38393 and SKF 81297 failed to exert such effects. On the other hand, the D2 antagonists raclopride and YM 09151-2, and D1 antagonists SCH 23390 and SKF 83566, all completely blocked d-amphetamine discrimination. The D2 antagonists produced more pronounced inhibitory effects on response rate than did D1 antagonists. Quinpirole substitution for d-amphetamine was blocked by YM 09151-2, but not by SCH 23390, while the locomotor stimulatory effect of quinpirole was inhibited by both drugs. The present findings confirm that D2 receptors play a primary role in the d-amphetamine discriminative cue, while the precise role of D1 receptors remains to be disclosed.

Animals↗
PubMed1991

N-methyl-D-aspartate increases the excitability of nigrostriatal dopamine terminals.

The terminal excitability of nigrostriatal dopamine cells was measured before and after i.v. administration of N-methyl-D-aspartate (NMDA; 1 or 4 mg/kg), the competitive NMDA antagonist AP-7 (2-amino-7-phosphonoheptanoic acid) (4 mg/kg) or saline. NMDA produced a dose-dependent increase in terminal excitability, in the absence of an effect on the somal membrane, whereas AP-7 and saline were without effect. These data provide convergent in vivo evidence that glutamate can enhance striatal dopamine release via the NMDA receptor.

2-Amino-5-phosphonovalerate↗
PubMed1991

Localization of [3H]GABA-labelled nerve fibre networks in the rat intestinal mucosa.

Laminar preparations of the rat colonic mucosa were treated for [3H]GABA (5 x 10(-8) M) autoradiography. Under conditions specific for high-affinity labelling of neuronal elements, a network of fibres overlying the base of the mucosal crypts and a second network coursing in close association with the crypts, were intensely labelled. Along the course of the overlying network of fibres, were dense accumulations of silver grains reminiscent of junctions for fibres projecting into the mucosa. All labelling could be prevented by the specific neuronal uptake inhibitor, L-DABA (10(-3) M).

Aminobutyrates↗
PubMed1991

Differential agonist profile of the enantiomers of 3-PPP at striatal dopamine autoreceptors: dependence on extracellular dopamine.

The effects of the enantiomers of 3-hydroxyphenyl-N-n-propylpiperidine (3-PPP) at dopamine (DA) synthesis modulating autoreceptors, measured as DOPA accumulation after decarboxylase inhibition, were assessed in vivo and in rat striatal slices. In vivo, (+)-3-PPP inhibited DOPA accumulation in the striatum, nucleus accumbens, and medial prefrontal cortex, whereas (-)-3-PPP either increased (striatal) or had no effect (accumbens, prefrontal cortex), on DOPA accumulation. In vitro, both (+)- and (-)-3-PPP reduced basal DOPA accumulation with a similar order of potency (apparent EC50 = 2.1 and 1.0 microns, respectively) and maximal effect, although they were less potent than the D2 DA receptor agonist quinpirole (EC50 = 0.15 microM). The inhibition of tyrosine hydroxylation was also observed in slices obtained from reserpine-pretreated rats and was blocked by the selective D2 DA antagonist (-)-sulpiride. This suggests that 3-PPP inhibition of DOPA accumulation was mediated directly by stimulation of DA D2 receptors. Increasing the amount of extracellular DA by depolarizing slices with 30 mM K+ did not alter the qualitative effects of either quinpirole or (+)-3-PPP. However, the stimulation of DA autoreceptors by (-)-3-PPP was no longer apparent under conditions of elevated extracellular DA. Under these depolarizing conditions, (-)-3-PPP actually antagonized the inhibitory effect afforded by either quinpirole or pergolide. A similar switch in profile was observed with transdihydrolisuride (TDHL). The data support the notion that (-)-3-PPP and TDHL are partial agonists at synthesis modulating DA autoreceptors.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1991

Effects of low, autoreceptor selective doses of dopamine agonists on the discriminative cue and locomotor hyperactivity produced by d-amphetamine.

The ability of low doses of the dopamine (DA) agonists quinpirole and (+)-3-PPP to reduce the discriminative stimulus properties and locomotor hyperactivity produced by d-amphetamine (0.5 mg/kg) was assessed in two groups of rats. Quinpirole (0.0125-0.05 mg/kg) and (+)-3-PPP (1.0-2.0 mg/kg) completely antagonized d-amphetamine-induced locomotor hyperactivity. In contrast, only single doses of quinpirole (0.025 mg/kg) and (+)-3-PPP (2.0 mg/kg) were effective in the drug discrimination paradigm; the antagonisms were small (18-47%), but significant. The inhibitory effects of quinpirole and (+)-3-PPP in these behavioural models are probably due to their ability to selectively stimulate DA autoreceptors in the nucleus accumbens and reduce the increase in DA release produced by d-amphetamine. It is suggested that the much weaker effects of the drugs in the discrimination paradigm are due to changes produced by the long-term periodic administration of d-amphetamine to these animals, such as a down-regulation in the sensitivity of DA autoreceptors.

Animals↗
PubMed1991

Behavioural profile of partial D2 dopamine receptor agonists. 1. Atypical inhibition of d-amphetamine-induced locomotor hyperactivity and stereotypy.

The effects of partial D2 dopamine (DA) receptor agonists on the behavioural activation produced by 1.5 and 8.0 mg/kg d-amphetamine were compared with the changes produced by the classical DA antagonist haloperidol. Alterations in behaviour were assessed in standard activity monitoring cages by direct observation of the rats using a rapid time sampling procedure. Haloperidol blocked d-amphetamine (1.5 mg/kg)-induced increases in photocell counts, ambulation, rearing and sniffing up, and after the highest dose of the DA antagonist the animals were mainly inactive. The partial D2 DA agonist SDZ 208-911 was equipotent to haloperidol in blocking the increase in photocell counts and rearing produced by d-amphetamine. However, even high doses of the drug did not reduce the incidence of sniffing or induce inactivity, but qualitative changes in the form of sniffing did occur. Although considerably less potent, preclamol exerted similar effects to SDZ 208-911. The profiles of SDZ 208-912 and terguride were intermediary to those of SDZ 208-911 and haloperidol. All compounds blocked the repetitive sniffing down produced by 8.0 mg/kg d-amphetamine. After a low dose of haloperidol, these stereotyped behaviours were replaced by a behavioural syndrome similar to that observed with low dose d-amphetamine, but inactivity was observed following a further small increase in antagonist dose. The blockade of stereotypy by SDZ 208-911, preclamol and terguride was accompanied only by the low dose d-amphetamine behavioural syndrome; no inhibition of sniffing or induction of inactivity occurred. SDZ 208-912 exhibited a profile with features very similar to that noted with haloperidol. These findings suggest that partial D2 agonists exert similar, but not identical, behavioural effects to classical DA antagonists when dopaminergic function in increased by d-amphetamine. The differences in behavioural profile are discussed in relation to variations in the intrinsic efficacy of the dopaminergic compounds and to differences in the response capability of D2 receptor populations underlying the different behaviours produced by d-amphetamine.

Animals↗
PubMed1991

The sequence of the Arbacia punctulata bindin cDNA and implications for the structural basis of species-specific sperm adhesion and fertilization.

Bindin is the major protein component of the acrosome granule of sea urchin sperm which mediates the species-specific adhesion of sperm to the egg surface during fertilization. Bindin isolated from both Arbacia punctulata and Strongylocentrotus purpuratus sperm demonstrate a distinct adhesive preference for eggs of the same species although a significant amount of cross-species reactivity is observed. Here we describe the isolation and sequence of A. punctulata bindin cDNA clones and a comparison of the predicted protein sequence with the sequence previously reported for S. purpuratus bindin (Gao et al., 1986, Proc. Natl. Acad. Sci., USA 83, 8634-8638). Bindins from these genera show substantial sequence similarity in both the mature bindin domain and the probindin precursor region. The most striking identity is a region of 42 conserved amino acids in the central part of the mature bindins. This conserved domain may be responsible for conserved functions of bindin. Regions flanking this conserved element on both the amino and carboxyl side are more highly divergent, suggesting that they are responsible for the species-specific properties of bindin. The mature A. punctulata sequence contains a putative transmembrane segment between residues 431 and 451 that is absent from S. purpuratus bindin. This structural element may account for the previous observation that isolated A. punctulata bindin uniquely forms multilamellar structures reminiscent of lipid bilayers and binds significant amounts of phospholipid and detergent. The structure of this hydrophobic segment also displays a number of similarities to viral fusion peptides.

Amino Acid Sequence↗
PubMed1991

High-affinity uptake of [3H]GABA by submucous ganglion cells, nerve fibres and peri- and para-vascular fibres in guinea-pig and rat intestine.

Segments of the intestinal wall from the guinea-pig and rat were dissected, and laminae from the submucous layer subjected to light microscopic [3H]GABA autoradiography. The laminae were carefully prepared so that their planar arrangement could be easily viewed. Intense labelling of fine processes by [3H]GABA was found in the different laminae of the submucous including Henle's or Schabadasch's plexus, the vascular nerve plexus, and a network of fibres subjacent to the muscularis mucosae. This labelling was extensive and represented neuronal-specific high-affinity uptake of radiolabelled GABA. The pattern of labelling was different between the laminae; however, within individual laminae, the distribution of labelled fibres was characteristic of the local nerve networks. Ganglia displayed intense labelling of neuropil and ganglion cells. These results provide strong evidence for the presence of GABAergic nerve cells and processes in the mammalian intestinal submucosa.

Animals↗