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D Smart

Publications and source records attributed to D Smart.

At least 91 records · Page 5Linked to original sources
PubMed1994

Characterisation of mu-opioid receptors on SH-SY5Y cells using naloxonazine and beta-funaltrexamine.

The irreversible opioid receptor antagonists naloxonazine and beta-funaltrexamine have been used to determine whether multiple mu-opioid receptors exist on undifferentiated SH-SY5Y human neuroblastoma cells. Naloxonazine binds irreversibly to the mu 1-opioid receptor subtype and reversibly to the mu 2-opioid receptor subtype. On SH-SY5Y cells naloxonazine afforded a Ki of 3.4 +/- 0.7 nM, and was fully reversible, indicating the mu-opioid receptor population on SH-SY5Y cells was solely of the mu 2-opioid receptor subtype. The alkylating agent beta-funaltrexamine was maximally able to alkylate only 60% of the mu-opioid receptor sites on SH-SY5Y cells, labelled with [3H]diprenorphine or [3H][D-Ala2,MePhe4,Gly(ol)5]enkephalin (DAMGO). The reversible binding of naloxonazine and the insensitivity of a percentage of the mu-opioid receptor sites to alkylation by beta-funaltrexamine suggests that differences do exist in the mu 2-opioid receptor population on undifferentiated SH-SY5Y cells. This may indicate further heterogeneity or the inability of beta-funaltrexamine to alkylate all relevant nucleophilic groups in a single population of receptors.

Amino Acid Sequence↗
PubMed1994

Halothane and isoflurane enhance basal and carbachol-stimulated inositol(1,4,5)triphosphate formation in SH-SY5Y human neuroblastoma cells.

The cellular mechanisms underlying the clinical effects of volatile anaesthetics remain unknown, although the plasma membrane and its associated proteins are likely targets. One such protein is the enzyme phospholipase C (PLC), which catalyses the formation of the second messenger inositol(1,4,5)triphosphate [Ins(1,4,5)P3]. Using SH-SY5Y human neuroblastoma cells we have demonstrated that halothane (0.50, 0.75 and 1.00%) enhances basal Ins(1,4,5)P3 mass formation approximately 1.8-fold. Halothane also caused a dose-dependent enhancement of carbachol-stimulated biphasic Ins(1,4,5)P3 formation at both the peak (half-maximal stimulation, EC50 = 0.76%) and plateau (EC50 = 0.74%) phases. At 1%, halothane did not alter the affinity for carbachol at either the peak (IC50: air = 9.4 +/- 1.5, halothane = 12.7 +/- 1.0 microM) or plateau (EC50: air = 11.7 +/- 1.2, halothane = 11.6 +/- 1.0 microM) phase, but did increase the maximum Ins(1,4,5)P3 response at both phases (air vs halothane: peak, 79.9 +/- 0.5 vs 124.8 +/- 2.5; plateau, 33.2 +/- 0.5 vs 47.9 +/- 0.6 pmol/mg protein). Isoflurane (2%) also enhanced basal and carbachol-stimulated Ins(1,4,5)P3 formation 2-fold and 1.5-fold, respectively. In summary, clinically relevant doses of the volatile anaesthetics halothane and isoflurane enhance basal and carbachol-stimulated Ins(1,4,5)P3 formation. Thus, activation of PLC, and subsequent potential Ins(1,4,5)P3-mediated rises in intracellular calcium, could play a part in the cellular mechanisms of volatile agent-induced anaesthesia.

Carbachol↗
PubMed1994

mu-Opioid receptor stimulation of inositol (1,4,5)trisphosphate formation via a pertussis toxin-sensitive G protein.

The cellular mechanisms underlying opioid action remain to be fully determined, although there is now growing indirect evidence that some opioid receptors may be coupled to phospholipase C. Using SH-SY5Y human neuroblastoma cells (expressing both mu- and delta-opioid receptors), we demonstrated that fentanyl, a mu-preferring opioid, caused a dose-dependent (EC50 = 16 nM) monophasic increase in inositol (1,4,5)trisphosphate mass formation that peaked at 15 s and returned to basal within 1-2 min. This response was of similar magnitude (25.4 +/- 0.8 pmol/mg of protein for 0.1 microM fentanyl) to that found in the plateau phase (5 min) following stimulation with 1 mM carbachol (18.3 +/- 1.4 pmol/mg of protein), and was naloxone-, but not naltrindole- (a delta antagonist), reversible. Further studies using [D-Ala2, MePhe4, Gly(ol)5]enkephalin and [D-Pen2,5]enkephalin confirmed that the response was specific for the mu receptor. Incubation with Ni2+ (2.5 mM) or in Ca(2+)-free buffer abolished the response, as did pretreatment (100 ng/ml for 24 h) with pertussis toxin (control plus 0.1 microM fentanyl, 26.9 +/- 1.5 pmol/mg of protein; pertussis-treated plus 0.1 microM fentanyl, 5.1 +/- 1.3 pmol/mg of protein). In summary, we have demonstrated a mu-opioid receptor-mediated activation of phospholipase C, via a pertussis toxin-sensitive G protein, that is Ca(2+)-dependent. This stimulatory effect of opioids on phospholipase C, and the potential inositol (1,4,5)trisphosphate-mediated rises in intracellular Ca2+, could play a part in the cellular mechanisms of opioid action.

Calcium↗
PubMed1994

Opioids and suckling in relation to inhibition of oestradiol-induced LH secretion in postpartum ewes.

The effects of time postpartum and suckling status on the oestradiol-induced LH surge in ewes and some of the possible mechanisms involved were investigated. Lambs were removed from nonsuckled ewes at parturition (day 0) and the ewes hand-milked 12 and 48 h later to prevent discomfort. Nonsuckled ewes on day 1, 7, 14 or 28, and suckled ewes on day 7 or 28 postpartum were injected i.m. with 50 micrograms oestradiol benzoate (hour 0). A further eight suckled ewes were given oestradiol on day 1 (as above), and four were also given naloxone (1 mg kg-1 h-1) from 10 until 14 h after administration of oestradiol benzoate. There was no LH surge in response to oestradiol on day 1. In nonsuckled ewes the onset of the LH surge was significantly (P < 0.05) delayed when oestradiol was administered on day 7 (17.4 +/- 1.5 h), compared with when it was injected on days 14 and 28 postpartum (14.4 +/- 1.7 and 14.0 +/- 1.4 h). The LH surge was also delayed (P < 0.05) when oestradiol was administered on day 7 (18.0 +/- 2.8 h) in suckled ewes. There was no difference in the duration or amplitude of the LH surge with time postpartum in nonsuckled ewes. Suckling significantly (P < 0.05) reduced the amplitude of the LH surge on both day 7 (21.0 +/- 2.4 versus 26.4 +/- 2.3 ng ml-1) and day 28 (23.6 +/- 3.2 versus 28.8 +/- 2.9 ng ml-1), but had no effect on the onset or duration of the surge.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1994

Transport stress delays the oestradiol-induced LH surge by a non-opioidergic mechanism in the early postpartum ewe.

The present study was designed to investigate whether transport, a mild environmental stressor, could affect the oestradiol-induced LH surge in postpartum ewes and, if so, the mechanism involved. Welsh Mountain ewes, with lambs removed at parturition (day 0) and hand-milked 12 and 48 h later, were given 50 micrograms oestradiol benzoate intramuscularly at various times postpartum. Blood samples were taken via an indwelling jugular venous catheter every 2 h from 8 to 24 h after oestradiol injection. All results are given as means +/- S.D. On day 1 oestradiol was unable to induce an LH surge in any ewe. Transport (10-14 h after oestradiol) delayed the onset of the oestradiol-induced LH surge on day 14 (17.5 +/- 1.7 vs 14.4 +/- 2.0 h, n = 5 each; P < 0.05), but not on day 28 (14.9 +/- 2.0 vs 14.0 +/- 2.4 h, n = 5 out of 7). Transport had no effect on the amplitude of the surge on either day. Naloxone treatment (1 mg/kg per 2 h) was unable to prevent the delay caused by transport (18.0 +/- 1.1 vs 17.5 +/- 1.7 h, n = 8 each), and did not affect the amplitude of the surge (28.4 +/- 5.3 vs 28.1 +/- 2.3 ng/ml, n = 8 each).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1993

Discussion paper: towards a systematic classification for regulatory peptides.

Advances in microsequencing technology have led to the elucidation of primary structures of peptidic messengers from many eukaryotic and prokaryotic life forms. Existing peptide nomenclature is based upon such factors as bioactivity, source of isolation (tissue or species), chemical attributes, acronyms derived from a combination of these factors, or by other arbitrary means. In order to overcome many of the problems arising from current nomenclature, a standardised classification scheme for peptidic messengers is proposed which utilises an alphanumeric code string analogous to EC enzyme classification to convey information on origin, chemistry and relatedness to other similar molecules. It is anticipated that the scheme outlined will provide the basis for the rational classification of new peptides.

Animals↗
PubMed1993

Use of specific antisera for the localisation and quantitation of leucokinin immunoreactivity in the nematode, Ascaris suum.

1. The leucokinins (LKs) are a group of eight related peptides isolated from the cockroach, Leucophaea maderae. 2. Antisera raised against LK-V, which were specific for the conserved mid to C-terminal region of the LKs, were used to immunostain the parasitic nematode Ascaris suum. 3. LK-IR was observed in neurons in the anterior nerve ring, retrovesicular ganglion, and ventral and dorsal nerve cords of the parasite. Immunostaining was specific in that it was abolished by preabsorption of the antiserum with different leucokinins. Some of the LK-immunoreactive neurons were identified on the basis of their morphological similarity with identified neurons in the free-living nematode Caenorhabditis elegans. 4. Immunoreactivity towards a number of other peptides, notably neuropeptide F, FMRF-amide, KGQELE and KELTAE, has previously been demonstrated in all LK-immunoreactive neurons, thus confirming the multiple-peptidergic nature of certain nematode nerves. 5. LK-IR was demonstrated and quantified in a number of tissues using RIA. Highest amounts were found in extracts of gut; LK-IR was also demonstrated in extracts of body wall, heads and tails, testes, ovaries and pseudocoelomic fluid. 6. The distribution of tissue LK-IR did not correlate with the amount of neuronal tissue in the samples. 7. Dilution experiments suggested that the LK-IR in the parasite is heterogeneous and that the peptide(s) in some tissues may not be analogous to the insect LKs. 8. The LK-IR in the parasite remains to be characterized; however, the results suggest that peptides related to the leucokinins may have a far wider phylogenetic distribution than has hitherto been thought.

Amino Acid Sequence↗
PubMed1993

Effect of needle changing and intravenous cannula collection on blood culture contamination rates.

STUDY OBJECTIVES: We tested the hypotheses that blood culture positivity and contamination rates were not increased by not changing needles between venipuncture and inoculation of blood culture bottles or by taking blood for culture by freshly inserted IV cannulae. DESIGN: A prospective study of blood cultures collected by venipuncture or IV cannulae taken from an emergency department population. Venipuncture samples were randomized into needle change (standard method) or no needle change before inoculation into blood culture bottles. PARTICIPANTS: Nine hundred forty patients requiring blood cultures after assessment in the ED. INTERVENTIONS: A standard disinfection procedure using 0.5% chlorhexidine in 70% alcohol was used. Blood was collected by venipuncture and inoculated with or without needle change. Blood collected by IV cannula was inoculated with a fresh needle applied to the collection syringe. MEASUREMENTS AND MAIN RESULTS: There was no statistically significant difference in contamination rates for blood collected by venipuncture with no needle change (6.4%) compared with needle change (4.2%, P > .30). No significant difference in contamination rates was noted for blood taken by freshly inserted IV cannulae (4.3%) compared with venipuncture with needle change after sampling (4.2%, P > .90). Some problems with randomization resulted in unequal numbers in the needle-change (286) versus no-needle-change (141) subgroups, and this may have introduced bias. A higher rate of pathogen growth was observed in blood taken by IV cannula (11.4%) compared with the standard method (6.3%) (P < .025). A significantly greater rate of Gram-negative sepsis was noted in the IV cannula group (6.6%) compared with direct venipuncture with needle change (1.1%) and no needle change (4.2%, P < .01). CONCLUSION: The results of this study do not support the practice of changing needles before inoculating blood samples into blood culture bottles. Collection of blood for culture through freshly inserted IV cannulae is associated with a low contamination rate and is an acceptable alternative to direct venipuncture. Sources of bias in this study suggest that further research is needed to determine the optimal technique for collecting blood cultures.

Bacteriological Techniques↗
PubMed1993

Immunocytochemical demonstration of neuropeptides in the central nervous system of the roundworm, Ascaris suum (Nematoda: Ascaroidea).

The localization and distribution of neuropeptides in the central nervous system of the pig roundworm, Ascaris suum, have been determined by an indirect immunofluorescence technique in conjunction with confocal microscopy. Antisera to 25 vertebrate peptides and two invertebrate peptides were used to screen the worm for immunoreactivity (IR). Immunostaining was obtained with antisera to pancreatic polypeptide (PP), peptide YY (PYY), neuropeptide Y (NPY), gastrin, cholecystokinin (CCK), substance P (SP), atrial natriuretic peptide (ANP), salmon gonadotropin-releasing hormone (SGnRH), mammalian gonadotropin-releasing hormone (MGnRH), chromogranin A (CGA) and FMRFamide. The most extensive patterns of IR occurred with antisera to PYY, FMRFamide and gastrin. IR was evident in nerve cells and fibres in the ganglia associated with the anterior nerve ring and in the main nerve cords and their commissures; IR to FMRFamide also occurred in the posterior nerve ring. Immunostaining for the other peptides was confined to the nerve cords, with the number of immunoreactive nerve fibres varying from peptide to peptide.

Animals↗
PubMed1993

Sex differences in psychological adjustment from infancy to 8 years.

The objective of this study was to explore sex differences in development from infancy to 8 years of age in a community sample. Measures of biological, social, interactive, and parental functioning as well as teacher reports were obtained. There were minimal differences in infancy, but major psychosocial differences emerged with increasing age. In the biological sphere boys were disadvantaged only in ratings of language and motor skills at 3 to 4 years old. They showed greater temperamental "difficulty" and low persistence factor scores from 5 years onward. Boys were significantly more likely to have problems with adaptive behavior and social competence and to show behavior problems of the hyperactive and aggressive type, as rated by mothers. Parent and family functioning measures did not differentiate between the sexes. Teachers rated boys as having more problems in academic and behavioral domains the first 3 years of school. Path analyses combining data sets gathered when the children were 3 to 8 years old demonstrated the differential courses of development for boys and girls although temperamental flexibility was the best predictor of behavioral adjustment for both sexes. A social learning explanation of the increased incidence of problems among males is supported, although biological influences are not ruled out.

Adaptation, Psychological↗
PubMed1993

Precursors of hyperactivity and aggression.

OBJECTIVE: This study investigated the early characteristics of children who at 8 years old were hyperactive, aggressive, or both, to clarify the etiology of these disorders. METHOD: Prospective data from infancy to 8 years on children who were hyperactive (N = 65), aggressive (N = 57), or both (N = 60) at 8 years were compared with those on a normal comparison group (N = 70). Variables included ratings of temperament, behavior, school performance, and socioeconomic and life stress indices. RESULTS: The two aggressive groups, particularly the hyperactive-aggressive group, were more difficult in temperament and behavior from infancy and had less optimal environments. In contrast, the "pure" hyperactive group showed more problems than did the comparison group only from 3 to 4 years on. All three clinical groups had poorer academic performance than did the controls. CONCLUSIONS: The findings are consistent with a transactional model of development in which aggression (with or without hyperactivity) emerges when difficultness in infancy interacts with a stressed environment. Hyperactivity, when unassociated with aggression, may emerge later from poor self-regulation when faced with societal, especially school, demands. The pattern of group differences found suggest that risk indicators for specific patterns of later maladjustment may be identifiable for early intervention.

Adolescent↗
PubMed1993

Repetitive nocturnal arterial oxygen desaturation and silent myocardial ischemia in patients presenting for vascular surgery.

OBJECTIVE: To determine whether nocturnal respiratory abnormality (cyclic oxygen desaturation and tachycardia) is associated with nocturnal myocardial ischemia in older individuals with ischemic heart disease. DESIGN: Non-invasive monitoring on a single occasion. SETTING: Tertiary care referral hospital. PATIENTS: Thirty four consecutive older (68.5 +/- 6 yrs) patients referred for elective abdominal or carotid reconstructive vascular surgery. RESULTS: Seven patients (21%) had moderately severe nocturnal respiratory abnormality, defined by more than 50 dips in arterial oxygen saturation and increases in heart rate during the night. Two of these seven had clinical risk factors for ischemic heart disease and had nocturnal myocardial ischemia. Ten patients (29%) developed ischemia at some time during the study, of whom seven hand known ischemic heart disease, hypertension, and/or angina. Those with increased nocturnal ischemia showed very low frequency (1-2 cycles per minute) cyclic heart rate oscillations and repetitive nocturnal episodes of arterial oxygen desaturation, similar to patients with sleep apnea. CONCLUSION: Repetitive nocturnal cyclic arterial desaturation and cyclic increases in heart rate are associated with nocturnal myocardial ischemia in individuals with clinical risk factors for ischemic heart disease. Further investigation in a large patient sample utilizing non-invasive monitoring of saturation, heart rate, and blood pressure may provide definitive evidence regarding causation of some of the nocturnal myocardial ischemia occurring in older individuals with vascular disease.

Aged↗
PubMed1992

The primary structure of TE-6: a novel neuropeptide from the nematode Ascaris suum.

Extensive immunoreactivity (IR) towards a hexapeptide (sequence KGQELE), which flanks the C-terminus of the pancreastatin sequence in rat chromogranin A (CGA), is found throughout the nervous system of the nematode parasite Ascaris suum. The peptide IR was purified from the gonoduct of the parasite and found to have the sequence TKQELE. This peptide, designated TE-6, has some C-terminal homology with several regions of the CGA molecule. However, TE-6 was the only peptide isolated suggesting that either the nematode does not possess CGA, or that the -ELE regions of parasite CGA-like peptides which would be larger than TE-6 are not accessible to the antiserum in RIA, or are not being successfully extracted from the parasite. The N-terminus of TE-6 has little homology with any of the sequences preceding -ELE regions in CGA. This, and the fact that the tissue from which TE-6 was isolated does not contain IR towards another, highly conserved, region of the CGA molecule (WE-14) suggests that TE-6 may belong to a new class of regulatory peptide unrelated to CGA.

Amino Acid Sequence↗
PubMed1992

Peptide tyrosine phenylalanine: a novel neuropeptide F-related nonapeptide from the brain of the squid, Loligo vulgaris.

A novel nonapeptide, sequence YAIVARPRFamide, was isolated from brain extracts of the squid, L. vulgaris. Designated peptide tyrosine phenylalanine (PYF), the peptide shows marked homology with the C-terminal nonapeptides of pancreatic polypeptide and neuropeptide F (NPF) from a number of sources. If PYF is the C-terminal nonapeptide of squid NPF, then it may be derived by a novel processing mechanism involving specific cleavage between two TYR residues. PYF may be a highly truncated, receptor-active variant of NPF.

Amino Acid Sequence↗