Biomedical subjects

L A Smith

Publications and source records attributed to L A Smith.

At least 127 records · Page 7Linked to original sources

PubMed1993

Cloning and functional expression of dendrotoxin K from black mamba, a K+ channel blocker.

Mamba dendrotoxins, 7K M(r) polypeptides with three disulfide bonds, selectively inhibit certain fast-activating, voltage-sensitive neuronal K+ channels and have been instrumental in their identification, localization, and purification. However, derivatives with more refined specificity are essential to define the structural and functional properties of the multiple subtypes known to reside in the nervous system. Hence, utilizing a constructed cDNA library from the venom glands of the black mamba (Dendroaspis polylepis), the gene encoding dendrotoxin K was isolated, amplified, and expressed as a maltose-binding fusion protein in the periplasmic space of Escherichia coli. After cleavage of the chaperone from the affinity-purified product, a recombinant protein was isolated and shown to be identical to native dendrotoxin K in its N-terminal sequence, chromatographic behavior, convulsive-inducing activity, and binding to voltage-activated K+ channels in bovine synaptic membranes. This successful expression of refolded active toxin, in adequate yield, makes possible for the first time the preparation of mutants with specificity tailored for each K+ channel subtype, based both on the recently derived three-dimensional structure of alpha-dendrotoxin and the identified binding site on cloned K+ channels.

Amino Acid Sequence↗

PubMed1993

Alterations in striatal and extrastriatal D-1 and D-2 dopamine receptors in the MPTP-treated common marmoset: an autoradiographic study.

In adult common marmosets (Callithrix jacchus), MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine) treatment induced almost total depletion of cells in the substantia nigra pars compacts (SNc) but partial cell loss in the ventral tegmental area (VTA). There was severe depletion of [3H]-mazindol binding to dopamine (DA) uptake sites in the caudate, putamen, and SNc. The loss of [3H]-mazindol binding in the nucleus accumbens (NAc) and olfactory tubercle (OT) was less marked. [3H]-mazindol binding in the body of caudate nucleus showed a small but significant recovery with increasing post-lesion survival times. The specific binding of [3H]-SCH 23390 to D-1 DA receptor sites was increased after MPTP treatment in all subregions of both caudate and putamen but was unaltered in the NAc and OT. Substantia nigra pars reticulata (SNr), frontal cortex, and medial segment of globus pallidus (GPm) all demonstrated moderate levels of [3H]-SCH 23390 binding in control animals, which were unaffected by MPTP treatment. Specific [3H]-spiperone binding to D-2 DA receptor sites was not altered by MPTP treatment in the subregions of caudate-putamen. Moderate levels of [3H]-spiperone binding were observed in control animals in the NAc, OT, SNc, and the lateral segment of globus pallidus (GP1). [3H]-spiperone binding in the SNc and OT was partially decreased in MPTP-treated animals. The changes in specific [3H]-spiperone and [3H]-SCH 23390 binding induced by MPTP-treatment did not alter with post-lesion survival times. These results demonstrate that MPTP treatment causes greater dopaminergic denervation of the caudate-putamen than in NAc/OT. This resulted in an increase in postsynaptic D-1 DA receptor sites in the caudate-putamen but not in the NAc/OT. Also, there appeared to be loss of presynaptic D-2 DA receptor sites in the SNc and OT. In the caudate-putamen, the loss of presynaptic D-2 DA receptor sites may have masked postsynaptic D-2 DA receptor upregulation.

1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine↗

PubMed1993

Assessing violent offending in serious juvenile offenders.

The convergent validity of the two most frequently used methods for assessing violent offending in juveniles (i.e., self-reports and arrests) was evaluated. Participants were 87 serious juvenile offenders and their maternal figures, primarily from disadvantaged families. Validation measures tapped established behavioral, family, and peer correlates of delinquency. Results failed to support the ability of either arrests for violent crimes or self-reported violent offenses to index violent criminal behavior accurately. Several methodological features of the study support our hypothesis that the findings were not spurious. Procedural and conceptual implications of the findings are discussed.

Adolescent↗

PubMed1993

Effects of the substance P receptor antagonist CP-96,345 on renal sensory receptor activation.

In anesthetized rats, we examined the effects of the substance P (SP) receptor antagonist CP-96,345 on the renorenal reflex responses to renal mechanoreceptor (MR) stimulation produced by increased ureteral pressure. Renal pelvic administration of SP at 0.16, 0.8, 4.0, 20, and 100 micrograms/ml increased ipsilateral afferent renal nerve activity (ARNA) in a concentration-dependent fashion, 45 +/- 28, 134 +/- 26, 185 +/- 24, 185 +/- 25, and 274 +/- 39%, respectively. Renal pelvic perfusion with CP-96,345 at 0.01, 0.1, and 1.0 mg/ml dose-dependently reduced the ARNA response to 4.0 micrograms/ml SP, by 5 +/- 17, 46 +/- 9, and 72 +/- 12%, respectively. Increasing ureteral pressure three times in the presence of renal pelvic perfusion with vehicle, 1 mg/ml CP-96,345, and vehicle, increased ARNA 77 +/- 21, 29 +/- 13, and 101 +/- 36%, respectively. Thus CP-96,345 produced a reversible blockade of the ARNA responses to renal MR stimulation. Increasing ureteral pressure three times in the presence of renal pelvic perfusion with vehicle, CP-96,345 (1 mg/ml), and CP-96,345 + 25-40 micrograms/ml SP, increased ARNA 108 +/- 15, 43 +/- 14, and 153 +/- 63%, respectively. Thus CP-96,345 produced a competitive blockade of the ARNA responses to renal MR stimulation. The contralateral diuretic and natriuretic responses to increased ureteral pressure were also blocked in a reversible and competitive fashion by CP-96,345. The inactive 2R,3R enantiomer of CP-96,345, CP-96,344 (1 mg/ml). had no effect on the ipsilateral ARNA or the contralateral renal excretory responses to increased ureteral pressure.(ABSTRACT TRUNCATED AT 250 WORDS)

Afferent Pathways↗

PubMed1993

Role of prostaglandins in renal sensory receptor activation by substance P and bradykinin.

In anesthetized rats increasing ureteral pressure results in an increase in ipsilateral afferent renal nerve activity and a reflex increase in contralateral urine flow rate and urinary sodium excretion that is dependent on intact prostaglandin synthesis. Activation of renal pelvic substance P receptors contributes to the renorenal reflex responses to increased ureteral pressure. Because these data suggested that renal sensory receptors could be activated by both prostaglandins and substance P we examined whether activation of renal sensory receptors by substance P was dependent on intact prostaglandin synthesis. The renal pelvis was perfused with capsaicin, 2.5 micrograms/ml, or substance P, 4 micrograms/ml, before and during renal pelvic perfusion with the prostaglandin synthesis inhibitor indomethacin, 50 micrograms/ml. Indomethacin reduced the peak ipsilateral afferent renal nerve activity responses to capsaicin and substance P by 83 +/- 15% and 81 +/- 8%, respectively, as well as the contralateral diuretic and natriuretic responses. We also examined the effects of renal pelvic administration of indomethacin on the responses to renal pelvic perfusion with bradykinin. Bradykinin, 20 micrograms/ml, increased peak ipsilateral afferent renal nerve activity by 197 +/- 47% and contralateral urine flow rate and urinary sodium excretion by 31 +/- 6 and 20 +/- 6%, respectively. Indomethacin reduced the ipsilateral afferent renal nerve activity response by 76 +/- 9% and abolished the contralateral diuretic and natriuretic responses to bradykinin. We conclude that renal sensory receptor activation by capsaicin, substance P, and bradykinin is dependent on intact renal prostaglandin synthesis.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1993

Effects of age at cordotomy and subsequent exercise on contraction times of motor units in the cat.

The contraction times (CTs) of functionally isolated motor units (MUs) in the soleus (SOL) and medial gastrocnemius (MG) muscles were determined in cats that had been spinalized at ages 2 (n = 15) or 12 (n = 9) wk and then either subjected to exercise on a treadmill or simply given manipulative care of the hindlimbs. The MUs were tested approximately 12 wk after the low-thoracic cordotomy, and comparisons were made with data from control animals. The CT of 50.9 ms obtained for SOL units (n = 163) in the spinal cats was 22% shorter than the mean of 65.0 ms for MUs (n = 57) from control cats (n = 4). Contrary to expectation, the CT in animals spinalized at 12 wk was significantly shorter than that in the 2-wk group. The CT for MG units (n = 105) in spinal cats was also significantly shorter (11%) than that in controls cats (n = 66, 6 cats), and those units identified by their high fatigue index as being of slow or fatigue-resistant type had a shorter CT than units with a low index. No distinction in CT of exercised and nonexercised groups was detected for either muscle. These findings are discussed in relation to the bearing influences of supraspinal and segmental origin have on CT duration in SOL and MG muscles during growth of the kitten. A slight, significant decrease (6%) in the fatigue index of SOL MUs (n = 144) was detected, but the values remained high (mean 0.87).

Aging↗

PubMed1992

High prevalence of human papillomavirus transcripts in all grades of cervical intraepithelial glandular neoplasia.

Cervical biopsy specimens containing cervical intraepithelial glandular neoplasia (CIGN) were examined for the presence of human papillomavirus (HPV) RNA transcripts by in situ hybridization with iodine 125-labeled riboprobes. HPV RNA was detectable in 95.2% of biopsy specimens. HPV 16 RNA was present in 12, HPV 18 in 27, and both in 1 of the 42 cases examined. Among HPV-positive cases, HPV RNA was detectable in all grades of CIGN and, in three cases, in glands displaying only minimal nuclear abnormality insufficient for a diagnosis of CIGN. Patients with HPV RNA-positive CIGN were younger than those with negative findings for HPV, and patients with less severe grades of CIGN showed a trend toward a younger age of presentation than patients with severe glandular lesions. Increasing grades of CIGN may reflect progressive stages in the development of cervical adenocarcinoma, and this progression may closely involve HPV gene expression from its earliest stages.

Adult↗

PubMed1992

Molecular cloning of the gene encoding developing seed L-asparaginase from Lupinus angustifolius.

A genomic sequence encoding Lupinus angustifolius L-asparaginase has been obtained, and is the first report of this gene from a plant source. The 3.2 kb of DNA sequenced contains a 1136 bp 5' flanking sequence, four exons and three introns. Intron-exon borders were mapped by comparing the genomic sequence with that of a L. arboreus cDNA. Primer extension analysis revealed transcription start sites 16 bp and 13 bp 5' of the initiating ATG for L. angustifolius and L. arboreus, respectively. The 5' flanking region contained sequences associated with seed-specific expression.

Amino Acid Sequence↗

PubMed1992

Molecular cloning of a cDNA encoding aspartate aminotransferase-P2 from lupin root nodules.

Two isoenzymic forms of aspartate aminotransferase are present in the plant fraction of developing lupin root nodules. One of these forms, aspartate aminotransferase-P2 (AAT-P2), increases dramatically with the onset of biological nitrogen fixation and is associated with the assimilation of ammonia by the plant in the Rhizobium-legume symbiosis. A day 18 lupin nodule cDNA library in the lambda ZapII vector was immunoscreened with a monoclonal antibody specific for AAT-P2 and yielded two near-full-length 1700 bp clones. These clones were sequenced. Amino acid sequences from three peptides derived from immunopurified AAT-P2 were aligned, and showed 100% homology with the amino acid sequence deduced from the cDNA clones. The DNA sequence showed 50% homology with AAT sequences from a range of animal sources. Conversion of the clones to the phagemid form allowed their expression in Escherichia coli where both exhibited enzyme activity that could be immunoprecipitated with AAT-P2-specific monoclonal antibodies. Western blot analysis revealed protein moieties with molecular masses of 39, 43, 45 and 55 kDa. The 5' end of the clones coded for a hydrophobic leader sequence of about 50 amino acids indicative of a targeting sequence and consistent with the plastid localisation of nodule AAT-P2.

Amino Acid Sequence↗

PubMed1992

Polymorphonuclear neutrophil leukocyte function in clinical bovine patients and in cows with or without Staphylococcus aureus mastitis.

A fluorochrome microassay was used to investigate peripheral blood polymorphonuclear leukocyte (PMNL) function in cattle. Glass-adherent PMNL were reacted with Staphylococcus aureus preincubated in 20% bovine serum for 30, 60 and 90 min. Coverslips were stained with acridine orange (AO) followed by crystal violet to quench extracellular bacterial fluorescence. PMNL function was evaluated by counting the number of dead (stained red with AO) and live (stained green with AO) S. aureus contained within 100 PMNL. A phagocytic index was calculated as the average number of bacteria contained within PMNL. The percentage killing of S. aureus was calculated from the average proportion of S. aureus within PMNL that were dead. Six clinically normal Holstein calves, 3-4 months of age, were sampled on 6 consecutive days. PMNL phagocytosis and killing did not vary significantly (p greater than 0.05) among repeated samplings per calf. PMNL function increased with increasing time of incubation of PMNL with S. aureus. Means (+/- SD) for percentage killing were 46.7 +/- 13.1, 57.4 +/- 11.6, and 62.1 +/- 9.8% for 30, 60 and 90 min of reaction, respectively. Means (+/- SD) for the phagocytic index were 2.9 +/- 0.8, 3.6 +/- 1.0, and 4.2 +/- 1.1 bacteria/PMNL for 30, 60 and 90 min of reaction, respectively. PMNL function was determined in 30 normal cattle of various breeds, age and sex, and these values were pooled to provide normal values for PMNL function. When values for bovine clinical patients (n = 25) with various diagnoses were compared with normal values (defined by the mean +/- 2SD for the 30 normal cattle) for PMNL function, only one patient was observed to exhibit PMNL hypofunction. A cow with disseminated intravascular coagulation in association with peracute coliform mastitis exhibited decreased PMNL killing capacity. Abnormal PMNL function was uncommon in the hospital population studied. Peripheral blood PMNL function was evaluated in lactating Holstein cows with (n = 15) or without (n = 15) chronic subclinical S. aureus mastitis. There was no significant (p greater than 0.05) difference in PMNL function among these cows.

Acridine Orange↗

PubMed1992

Motor activity following the administration of selective D-1 and D-2 dopaminergic drugs to MPTP-treated common marmosets.

The ability of selective D-1 agonist and antagonist drugs to alter motor deficits and locomotor activity was studied in MPTP-treated common marmosets. Both the D-2 agonist quinpirole and the mixed D-1/D-2 agonist apomorphine reversed the motor impairments and induced locomotor activity. The D-1 antagonist SCH 23390 and the D-2 antagonist raclopride given alone further reduced motor function in MPTP-treated animals. The actions of quinpirole were potently and completely inhibited by raclopride but only partially and inconsistently by SCH 23390. In contrast, the effects of apomorphine were markedly but incompletely inhibited by both raclopride and SCH 23390. The D-1 agonist SKF 38393 alone caused a dose related reduction in motor activity. SKF 38393 weakly and partially inhibited the improvements in motor function produced by quinpirole but had a more pronounced effect on apomorphine induced motor activity. The induction of motor activity in MPTP treated common marmosets may separately involve both D-1 and D-2 receptors. Comparison with our previous data on the effect of the same drugs in normal common marmosets provides some evidence for a breakdown of linkage between D-1 and D-2 systems following MPTP treatment. The actions of SKF 38393 in MPTP-treated common marmosets contrasts with its ability to induce behavioural activation and a facilitation of D-2 mediated behaviour in rodents. SKF 38393 may not be the compound with which to delineate the role of D-1 receptors in primates.

1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine↗

PubMed1992

Molecular properties and structure-function relationships of lethal peptides from venom of Wagler's pit viper, Trimeresurus wagleri.

Two new lethal peptides (waglerins) were purified from the venom of Trimeresurus wagleri, and sequenced. We found them to be analogs of lethal peptides (waglerins) I and II reported previously (Weinstein et al., Toxicon 29, 227-236, 1991), with an additional Ser-Leu on the amino terminus. Three of the four waglerins were synthesized and the products were chemically and biologically equivalent to the naturally occurring counterparts in venom. Murine i.p. LD50 for synthetic waglerins I, SL-I and II were 0.33, 0.22, and 0.51 mg/kg, respectively. The single, intramolecular disulfide bond in each synthetic peptide formed rapidly in high yield. The reduced (cysteine-containing) forms of the peptides appeared to have significant toxicities, even without prior disulfide bond formation, but synthetic analogs with serine substituted for cysteine were not toxic. The synthetic dimer of waglerin I, formed by two intermolecular disulfide bonds, was not toxic, but rapidly rearranged to lethal, monomeric waglerin I at alkaline pH upon the addition of 5 mM beta-mercaptoethanol. Waglerin I was inactivated by cleavage at Tyr-15 with chymotrypsin.

Amino Acid Sequence↗

PubMed1992

Pathologic changes induced by phospholipase A2 isolated from the venom of Collett's snake, Pseudechis colletti: a light and electron microscopic study.

Pseudechis colletti is an Australian elapid snake with a range limited to central Queensland, Australia. The venom of this snake, as well as that of several other Australian elapids, has been shown to contain a phospholipase A2 (PLA2) which can cause a marked myoglobinuria in mice. Few studies have described the histopathologic and ultrastructural changes that result from myotoxic PLA2-induced damage. Our investigation demonstrated that the isolated PLA2 induced myodegeneration and necrosis in myocardium in a dose-related manner, with subsequent myoglobinuria and myoglobinuric nephropathy.

Animals↗

PubMed1992

Preliminary evidence for a postsynaptic action of beta-bungarotoxin in mammalian skeletal muscle.

Two hours after treatment with beta-bungarotoxin (0.34-0.4 microM), when there was complete neuromuscular block, the peak contracture response to 50 microM succinylcholine was significantly reduced by about 35% in the mouse phrenic nerve-diaphragm preparation. Additionally, significant phospholipase A2 activity was detected on primary cell cultures from skeletal muscle which were incubated for 2 hr with concentrations of beta-bungarotoxin greater than or equal to 0.1 microM. Thus, beta-bungarotoxin appears to have pharmacologically and biochemically detectable postsynaptic actions in mammalian muscle systems.

Animals↗

PubMed1992

Family preservation using multisystemic therapy: an effective alternative to incarcerating serious juvenile offenders.

Multisystemic therapy (MST) delivered through a community mental health center was compared with usual services delivered by a Department of Youth Services in the treatment of 84 serious juvenile offenders and their multiproblem families. Offenders were assigned randomly to treatment conditions. Pretreatment and posttreatment assessment batteries evaluating family relations, peer relations, symptomatology, social competence, and self-reported delinquency were completed by the youth and a parent, and archival records were searched at 59 weeks postreferral to obtain data on rearrest and incarceration. In comparison with youths who received usual services, youths who received MST had fewer arrests and self-reported offenses and spent an average of 10 fewer weeks incarcerated. In addition, families in the MST condition reported increased family cohesion and decreased youth aggression in peer relations. The relative effectiveness of MST was neither moderated by demographic characteristics nor mediated by psychosocial variables.

Adolescent↗

PubMed1991

Relation of blood pressure to cognitive function in the elderly.

Clinical case-control studies of the relation between blood pressure and cognitive function have generally found lower function among hypertensives. Most of these studies were small and incompletely controlled for confounders. Two population-based studies have yielded conflicting results. This study examines cognitive function over the entire range of blood pressure in a defined elderly population. A questionnaire administered in the home to 3,809 persons aged greater than or equal to 65 years in East Boston, Massachusetts, in 1982 and 1983 contained four brief cognitive tests: immediate memory, delayed memory, a mental status questionnaire, and digit span. In linear regression analyses adjusting for age, sex, and education, the direction of the association was not consistent among the tests. An increase in diastolic pressure of 10 mmHg was associated with an increase of 1.0 in percentile scores on the immediate memory test (95% confidence interval (CI) -0.04 to 1.9); with an increase of 1.1 in percentile scores on the delayed memory test (95% CI -0.1 to 2.3); with a decrease of -0.8 in percentile scores on the mental status questionnaire (95% CI -1.8 to 0.2); and with a decrease of -0.9 in percentile scores on the attention test (95% CI -1.5 to -0.2). These results suggest that blood pressure is not a substantial contributor to cognitive status in the elderly.

Aged↗

PubMed1991

Risk of death from Alzheimer's disease in a community population of older persons.

A random sample of 467 persons over age 65 years from the population of an urban US community, stratified by age, sex, and performance on a brief memory test, underwent clinical evaluation for dementing illness in 1982-1984. Of these persons, 134 had probable Alzheimer's disease, 166 had possible Alzheimer's disease, and 167 had no evidence of Alzheimer's disease. Over a median follow-up period of 4.9 years following evaluation, 165 (35%) died. Overall, persons with probable Alzheimer's disease had a relative risk of death 1.44 (95% confidence interval (Cl) 1.05-1.96) times that of the unaffected. Level of cognitive impairment and the presence of cachexia upon physical examination both strongly and independently modified risk of death. Among those with probable Alzheimer's disease, mortality for those with mild or moderate cognitive impairment and no evidence of cachexia was comparable to that of the unaffected. However, among those with probable Alzheimer's disease and either severe cognitive impairment or cachexia, the risk of death was substantially higher. Persons with probable Alzheimer's disease who had both severe cognitive impairment and clear cachexia had a risk of death 4.60 (95% Cl 1.63-13.1) times that of unaffected persons.

Aged↗

PubMed1991

Motor activity following the administration of selective D-1 and D-2 dopaminergic drugs to normal common marmosets.

In normal common marmosets administration of the D-1/D-2 agonist apomorphine or the selective D-2 agonist quinpirole caused a dose-dependent increase in motor activity and induced stereotyped behaviour. Both the selective D-2 antagonist raclopride and the selective D-1 antagonist SCH 23390 inhibited normal locomotor activity and induced catalepsy. Quinpirole- and apomorphine-induced motor activity were potently inhibited by pretreatment with raclopride. The effects of quinpirole, but not apomorphine, were weakly inhibited by SCH 23390. The selective D-1 partial agonist SKF 38393 decreased motor activity and did not induce grooming, oral movements or other behaviours. SKF 38393 inhibited motor activity induced by the administration of quinpirole but did not alter apomorphine-induced motor behaviour. Locomotor activity in normal common marmosets appears to be mediated mainly via D-2 systems. In contrast to rodents, administration of SKF 38393 does not induce behavioural activation and there does not appear to be a facilitating effect of D-1 systems on D-2 function in the normal common marmoset. However, the ability of both SKF 38393 and SCH 23390 to inhibit quinpirole locomotor activity suggests some interaction between D-1 and D-2 systems to occur in this species.

2,3,4,5-Tetrahydro-7,8-dihydroxy-1-phenyl-1H-3-ben↗