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Biomedical subjects

D Lee

Publications and source records attributed to D Lee.

At least 361 records · Page 20Linked to original sources
PubMed1995

COOH-terminal disruption of lipoprotein lipase in mice is lethal in homozygotes, but heterozygotes have elevated triglycerides and impaired enzyme activity.

The role of the enzyme lipoprotein lipase (LPL) in atherosclerosis is uncertain. To generate an animal model of LPL deficiency, we targeted the LPL gene in embryonic stem cells with a vector designed to disrupt the COOH terminus of the protein and used these cells to generate LPL-deficient mice. Germ line transmission of the disrupted LPL allele was achieved with two chimeric males, and offspring from each of these animals were phenotypically identical. Pups homozygous (-/-) for LPL deficiency died within 48 h of birth with extreme elevations of serum triglycerides (13,327 mg/dl) associated with essentially absent LPL enzyme activity in heart and carcass. Newborn heterozygous (+/-) LPL-deficient pups had lower LPL enzyme activity and higher triglycerides (370 versus 121 mg/dl) than wild type (+/+) littermates. Adult heterozygotes had higher triglycerides than wild type mice with ad libitum feeding (236 mg/dl for +/- versus 88 mg/dl for +/+) and after fasting for 4 h (98 mg/dl for +/- versus 51 for +/+) or 12 h (109 mg/dl for +/- versus 56 mg/dl for +/+). Triglycerides were present as very low density lipoprotein particles and chylomicrons, but high density lipoprotein cholesterol levels were not decreased in +/- animals. Plasma heparin-releasable LPL activity was 43% lower in +/- versus +/+ adult animals. LPL activity, mRNA, and protein were lower in the tissues of +/- versus +/+ mice. Homozygous LPL deficiency caused by disruption of the COOH terminus of the enzyme is lethal in mice. Heterozygous LPL deficiency caused by this mutation is associated with mild to moderate hypertriglyceridemia without affecting static HDL cholesterol levels. Heterozygous LPL-deficient mice could be useful for determining if hypertriglyceridemia, independently or in combination with other discrete defects, influences atherosclerosis.

Animals↗
PubMed1995

Does ulnar neuropathy predispose to focal dystonia?

We have observed a high incidence of ulnar neuropathy in musicians with dystonic flexion of the ipsilateral little and ring fingers. To investigate the relationship between ulnar neuropathy and focal dystonia, we compared the patterns of surface EMG activity in extensor digitorum communis (EDC4) and flexor digitorum superficialis (FDS4) during tapping of the ring finger in normal controls and patients with ulnar neuropathy or focal dystonia. Ten of 10 normal subjects exhibited well-formed alternating EMG bursts in EDC4 and FDS4 separated by clear silent periods. Seven of 7 patients with dystonic flexion of the little and ring fingers showed loss of silent periods between poorly formed bursts in FDS or EDC. Surprisingly, 9 of 10 patients with ulnar neuropathy showed burst pattern abnormalities qualitatively similar to those observed in the dystonic patients. These data suggest that ulnar neuropathy alters the execution of a motor task involving multiple peripheral nerves.

Adult↗
PubMed1995

The Arabidopsis thaliana 4-coumarate:CoA ligase (4CL) gene: stress and developmentally regulated expression and nucleotide sequence of its cDNA.

An Arabidopsis cDNA clone encoding 4-coumarate:CoA ligase (4CL), a key enzyme of phenylpropanoid metabolism, was identified and sequenced. The predicted amino acid sequence is similar to those of other cloned 4CL genes. Southern blot analysis indicated that 4CL is single-copy gene in Arabidopsis. Northern blots showed that 4CL expression was activated early during seedling development. The onset of 4CL expression was correlated with the onset of lignin deposition in cotyledons and roots 2-3 days after germination. The timing of the expression of a parsley 4CL1-GUS fusion in transgenic Arabidopsis seedlings was examined in parallel and was very similar to that of endogenous 4CL. In mature plants, highest 4CL expression was observed in bolting stems, where relatively large amounts of lignin accumulate. Both 4CL and 4CL1-GUS mRNA accumulation was strongly and transiently activated by wounding of mature Arabidopsis leaves. 4CL expression was specifically activated within 6 h after infiltration of Arabidopsis ecotype Columbia leaves with a Pseudomonas syringae pv. maculicola strain harboring the bacterial avirulence gene avrB, which causes in incompatible interaction. The timing of 4CL activation was identical to the previously observed activation of PAL gene expression in this interaction. No activation of 4CL expression was observed in a compatible interaction caused by a Pseudomonas syringae pv. maculicola strain without avrB.

Amino Acid Sequence↗
PubMed1995

Effect of selenium depletion on thyroidal type-I iodothyronine deiodinase activity in isolated human thyrocytes and rat thyroid and liver.

The effects of dietary selenium deficiency on hepatic and thyroidal type I iodothyronine deiodinase (ID-I) and selenium-dependent glutathione peroxidase (GPx) activities have been studied in weanling rats. In selenium-deficient animals hepatic ID-I activity was reduced to 11% of the activity found in the selenium-replete groups, whilst thyroidal ID-I activity increased by 42%. Hepatic and thyroidal GPx activities were also reduced by selenium deficiency to approximately 0.6 and 70%, respectively, of the values found in the selenium-replete animals. We have also studied the effects of thyrotropin (TSH), and selenium supply on the activity of IDI and GPx in human thyrocytes grown in primary culture. When thyrocytes were grown in selenium-deficient (< 1 nmol l-1 Se) medium in the absence of TSH, addition of sodium selenite up to 1000 nmol l-1 had little or no effect on ID-I activity. In the absence of added selenite, TSH addition produced a significant increase in ID-I activity and this stimulation was increased further when selenite was added at concentrations of 50-1000 nmol l-1 with an optimal effect on ID-I activity being observed at a 500 nmol l-1. Selenium content and GPx activity in human thyrocytes grown in selenium-free media (selenium content < 1 nmol l-1) were not significantly lower than the corresponding measurements made in cells grown in media containing selenium at a concentration of 5.4 nmol l-1.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1995

Localized Mycobacterium avium-intracellulare mastitis in an immunocompetent woman with silicone breast implants.

While M. avium-intracellulare has received increasing recognition due to its association with AIDS, silicone breast implants have come under increased FDA scrutiny, requiring new safety and efficacy protocols. We report a case of localized M. avium-intracellulare mastitis associated with a breast implant in an immunocompetent, HIV-negative woman who was cured with implant removal and a long course (6 months) of clarithromycin. Serous exudates occurring in association with breast implants should be cultured for acid-fast organisms. Clarithromycin may be a therapeutic adjunct to effectively cure these infections.

Adult↗
PubMed1995

Multicentre trial of antepartum low-dose anti-D immunoglobulin.

Routine antenatal Rh immunoprophylaxis would substantially increase the use of anti-D Ig in the U.K. As availability of anti-D Ig is one factor influencing a decision to introduce routine antenatal prophylaxis, a trial was undertaken to test the efficacy of a lower dose of anti-D Ig than that used in earlier studies. RhD-negative primigravidae were randomized as controls or recipients of two doses of 250iu of anti-D Ig given at 28 and 34 weeks gestation. Blood samples were tested at delivery and at 6 months postpartum for the presence of immune anti-D, and again later if results were equivocal. Nine (1.5%) out of 595 control patients had immune anti-D at follow-up at 6 months and later; 4 (0.78%) of 513 treated women were immunized. It was concluded that, while two doses of 250iu of anti-D Ig may reduce alloimmunization, they are not as effective as two doses of 500iu in a previous trial.

Dose-Response Relationship, Drug↗
PubMed1995

Quantification of anti-c in haemolytic disease of the newborn.

Anti-c is an important Rh antibody that causes haemolytic disease of the newborn (HDN). We have carried out a retrospective analysis of the clinical outcome of pregnancy in 120 mothers with anti-c. Of these, 100 gave birth to c-positive infants, of whom 14 had severe HDN requiring exchange transfusion. In all of these, the maternal level was 9.5 iu/ml or greater. Of the 29 women with anti-c levels of 9.5 iu/ml or above, in addition to the 14 with severely affected infants, 15 had infants requiring only phototherapy or no treatment. Our observations suggest that when the anti-c level is below 7.5 iu/ml, the fetus is unlikely to be seriously affected and invasive obstetric intervention is unnecessary. Of the 120 women studied, 50% had had a blood transfusion, in most cases for obstetric complications in a previous pregnancy. Although it was not possible to attribute alloimmunization to blood transfusion rather than previous pregnancy in any individual case, this observation points to the value of routine c typing as part of antenatal screening, so that c-negative blood can be selected.

Bilirubin↗
PubMed1995

Anti-D quantification by flow cytometry: an alternative to the AutoAnalyser?

A flow cytometry method for quantifying levels of anti-D is described. The method is based on the indirect antiglobulin test and uses an FITC Fab anti-human IgG reagent for detection of bound anti-D. A standard curve is generated from the fluorescence measured by flow cytometry using the British Standard anti-D preparation 73/515. The fluorescence obtained from test samples by flow cytometry is converted into IU/mL anti-D, using the standard curve. In a limited study with 42 serum samples the assay showed good correlation with results obtained using the AutoAnalyser for sera with anti-D levels less than 100 IU/mL. The method is simple to perform and has potential as a replacement for the AutoAnalyser.

Autoanalysis↗
PubMed1995

An open trial of exposure therapy based on deconditioning for post-traumatic stress disorder.

Twenty-three patients who had experienced a major stressful event were given a debriefing session followed by eight weekly sessions of imaginal exposure and in vivo exposure. Patients recounted their traumatic experiences aloud, using the first person and the present tense, and included as much detail as possible. This account was audiotaped and patients were asked to listen to the tape between treatment sessions. There were reductions of 42 percent in the Impact of Events Scale (IES), of 61 percent in the General Health Questionnaire (GHQ), of 38 percent in the Symptom Checklist-90 (SCL-90) questionnaire, and of 35 percent in the Clinician Administered Post-Traumatic Stress scale (CAPS), all of which were statistically significant. The number of patients who satisfied the diagnostic criteria for post-traumatic stress disorder was halved.

Adult↗
PubMed1995

Flow cytometry in diagnosis and management of large fetomaternal haemorrhage.

AIMS: To evaluate an indirect immunofluorescence flow cytometry technique in a series of patients with large fetomaternal haemorrhage (FMH). METHODS: Patient samples identified by Kleihauer testing in local laboratories as having FMH > 4 ml were sent for flow cytometric analysis. In a proportion of cases the mothers received anti-D immunoglobulin prophylaxis according to the flow cytometer estimate of FMH volume. RESULTS: Forty three cases of FMH were studied prospectively. The correlation between Kleihauer and flow cytometry results was poor. In 38 (88%) cases the size of FMH quantitated by flow cytometry was lower than that estimated using the Kleihauer technique. In 13 (30%) cases no Rh D immunoglobulin positive cells were detected by flow cytometry. Centralised review of the original Kleihauer films using a calibrated microscope resulted in improved, but still suboptimal correlation with flow cytometry results. In 15 cases anti-D immunoglobulin was given according to the flow cytometer estimation of FMH size, resulting in a 58% reduction in the amount of anti-D immunoglobulin given. None of the patients were immunised when tested six months later. CONCLUSIONS: Flow cytometry is helpful for the accurate quantitation and management of patients with large FMH and in cases where the presence of maternal haemoglobin F containing cells renders the Kleihauer technique inaccurate. Worthwhile reductions in the use of anti-D immunoglobulin can be achieved.

Cell Separation↗
PubMed1995

Glucocorticoids inhibit D1B, but not D2, receptor-mediated effects on hypothalamic atrial natriuretic factor neurons.

Recent evidence suggests that ANF neurons of the hypothalamus are dopamine sensitive, and the catecholamine may exert a direct stimulatory or inhibitory effect on the neurons mediated through D1 or D2 receptors, respectively, in a manner related to the differential dopamine binding sensitivity of the two receptor subtypes. Employing well characterized ANF RIA and colorimetric Northern blot analysis with synthetic oligonucleotide probes complementary to pro-ANF messenger RNA (mRNA), we report here the effect of dexamethasone (DM), a potent synthetic glucocorticoid, on DA-stimulated ANF neurons in long term primary cultures of neonatal rat hypothalamic cells. Although DM alone did not affect basal secretion of immunoreactive ANF, it approximately halved immunoreactive ANF secretion induced by D1 agonist, SKF38393 (P < 0.01). The effect of DM was both time dependent and dose related, with an EC50 of 0.1 nM; it was blocked by 100 nM RU38486 (P < 0.05), a glucocorticoid receptor antagonist, but not by 100 nM RU28318, a mineralocorticoid receptor antagonist. In addition, the effect of DM was mimicked by corticosterone (EC50, 10 nM), but not deoxycorticosterone. The increased expression of pro-ANF mRNA signal induced by the D1 agonist in culture was suppressed by DM in a similar manner. In contrast, DM did not modulate ANF production and secretion induced by D2 agonist, quinpirole. Furthermore, reverse transcription-polymerase chain reaction demonstrated that D1B, but not D2, receptor mRNA expression was selectively suppressed by glucocorticoids. Thus, we conclude that in monolayer cultures of rat hypothalamic neurons, glucocorticoids differentially modulate dopamine receptor-induced responsiveness of ANF neurons by down-regulating D1B, but not D2, receptor-mediated changes. Hence, in severe stress, high levels of circulating glucocorticoids may negate the D1B-induced stimulatory response but allow dopamine to suppress the function of hypothalamic ANF neurons through D2 receptor activation.

Animals↗
PubMed1995

D1 receptors mediate dopamine stimulation of immunoreactive atrial natriuretic factor (ANF) release and pro-ANF messenger ribonucleic acid expression of rat hypothalamic neurons in culture.

Atrial natriuretic factor (ANF) and its smaller congeners are produced and released from rat brains to regulate the cardiovascular system, drinking behavior, and neurohormone release at the central level. In the hypothalamus, the adenylyl cyclase-cAMP system plays an important role in modulating the function of ANF neurons in the paraventricular nuclei and the periventricular regions that receive rich dopaminergic inputs. We report here a novel observation of a dopamine (DA) D1 agonist, SKF-38393, modulating immunoreactive (ir) ANF and pro-ANF messenger RNA (mRNA) expression in rat hypothalamic neurons. In long term primary cultures of neonatal rat hypothalamic cells, treatment with SKF-38393 increased irANF secretion in a time-related and dose-dependent manner, with an ED50 of approximately 10(-7) M and a concentration producing maximum effect of 10(-5) M. This stimulating effect of SKF-38393 was mimicked by 10(-5) M DA, a physiological ligand for D1 receptors. Furthermore, both of the stimulatory effects were abolished by SCH-23390, a D1 antagonist. These immunoassay findings were accompanied by corresponding changes in the abundance of pro-ANF mRNA in the cultures, as examined by colorimetric Northern blot analysis. By combining the techniques of in situ hybridization and immunocytochemistry, the mRNA of D1 receptor was colocalized in approximately 90% of the irANF-positive cells in the cultures. In addition, cholera toxin, an irreversible activator of adenylyl cyclases, markedly increased irANF secretion and cAMP production in a dose-dependent manner. This effect mimicked that of D1 agonist-stimulated ANF release, in that the latter also concurrently enhanced cAMP production in the hypothalamic cultures. Furthermore, the stimulatory effect of D1 agonist on irANF release was markedly suppressed by rp-cAMPS, a cAMP antagonist. We, thus, conclude that the release and gene expression of ANF in rat hypothalamic neurons are directly stimulated by DA acting through its D1 receptors on ANF neurons; this effect may operate at the genomic level and is mediated at least in part through activation of the cAMP-dependent kinase-A pathway.

2,3,4,5-Tetrahydro-7,8-dihydroxy-1-phenyl-1H-3-ben↗
PubMed1995

Elevation of microtubule-associated protein tau in the cerebrospinal fluid of patients with Alzheimer's disease.

Currently, there is no biochemical marker clinically available to test for the presence of Alzheimer's disease (AD). Recent studies suggest that the core component of AD-associated neurofibrillary tangles (NFTs), the microtubule-associated protein tau, might be present in CSF. This study focuses on establishing both the presence of tau in CSF and its potential utility in the diagnosis of AD. We obtained CSF from 181 individuals; 71 of these were diagnosed as having probable AD by NINCDS-ADRDA criteria. The remaining 110 individuals were divided into three groups: (1) age-matched demented non-AD patients (n = 25), (2) neurologic controls (n = 59), and (3) other controls (n = 26). We developed a sensitive enzyme-linked immunosorbent tau assay using monoclonal antibodies prepared against recombinant human tau. We confirmed specificity of the antibodies by a combination of immunoprecipitation and immunoblot results. By this assay we measured that the AD population has a mean level of tau 50% greater than the non-AD dementia patients. Comparing AD patients with all other groups, the difference in tau levels as analyzed by one-way ANOVA is highly statistically significant (p < 0.001). Postmortem analysis of two AD patients with high levels of CSF tau revealed a high density of NFTs in the hippocampus. There was no significant correlation between tau and age in the non-AD groups. This study suggests that CSF tau is elevated in AD and might be a useful aid in antemortem diagnosis.

Aged↗