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Biomedical subjects

M Miller

Publications and source records attributed to M Miller.

At least 433 records · Page 24Linked to original sources
PubMed1993

Membrane topology and omeprazole labeling of the gastric H+,K(+)-adenosinetriphosphatase.

The gastric H+,K(+)-ATPase is an alpha beta heterodimer with close homology to the Na+,K(+)-ATPase. Digestion of intact cytoplasmic-side-out vesicles at a trypsin to protein ratio of 1/4 removed most of the cytoplasmic protein, leaving membrane-spanning pairs in high yield. These were visualized on gels and poly(vinylidene difluoride) (PVDF) membranes by sodium dodecyl sulfate solubilization of the membrane-embedded segments and labeling of the cysteine residues with fluorescein maleimide prior to electrophoresis. The membrane-spanning residues of the alpha subunit were found between positions 104 and 162 (M1/M2), 291 and 358(M3/M4), 776 and 835 (M5/M6), and 853 and 946 (M7/M8). Although this method did not detect membrane retention of the hydrophobic sequences subsequent to position 946, it provided biochemical evidence for at least eight membrane segments in the catalytic subunit. Intact vesicles containing this enzyme transport acid in the presence of KCl, valinomycin, and MgATP. Omeprazole accumulates in these acidified vesicles and converts to a cationic sulfenamide. This forms disulfides with accessible cysteines. The reaction with this extracytoplasmic thiol reagent inhibits ATPase activity. Full inhibition was obtained with a stoichiometry of 2.2 mol of omeprazole bound/mg of protein. Only the alpha subunit was labeled. The cysteines reacting with omeprazole were defined by proteolytic cleavage of 3H- or 14C-omeprazole-labeled enzyme followed by peptide sequencing of fragments separated on tricine gradient gels and transferred to PVDF membranes. Tryptic digestion at a 1/40 trypsin to protein ratio in the presence of ligands that stabilize the E2P form of the enzyme produced two large fragments, one of 68 kDa stretching from Glu47 to probably Arg666 that contained minor labeling and the other of 333 kDa beginning at Ala671 and extending to probably Arg946 that contained greater than 85% of the label. Digestion of labeled vesicles at 1/75 or 1/4 trypsin to protein ratios gave radioactive patterns consistent with labeling at Cys813 and/or Cys822 and at Cys892 and/or Cys927 and/or Cys938. V8 protease digestion of the solubilized alpha subunit produced a fragment extending from Ser838 to possible Asp900 that was omeprazole-labeled, showing that Cys892 was labeled and Cys927 and Cys938 were not. Hence, omeprazole labels the H+,K(+)-ATPase at cysteines within the M5/M6 and M7/M8 regions of the alpha subunit, accounting for its inhibitory action in vivo and in vitro.

Adenosine Triphosphate↗
PubMed1993

Alteplase versus heparin in acute pulmonary embolism: randomised trial assessing right-ventricular function and pulmonary perfusion.

Data from a non-randomised study have hinted that in patients with acute pulmonary embolism (PE), thrombolysis followed by heparin more rapidly reverses right-ventricular dysfunction and restores pulmonary tissue perfusion than does heparin alone. We have pursued this idea in a randomised protocol. 46 haemodynamically stable patients were randomised to recombinant tissue plasminogen activator (alteplase, rt-PA) 100 mg over 2 h followed by intravenous heparin and 55 to heparin alone. Right-ventricular wall motion was assessed qualitatively, and right-ventricular end diastolic area was estimated by planimetry from echocardiograms at baseline and at 3 and 24 hours. Pulmonary perfusion scans were obtained at baseline and 24 hours. In 39% of rt-PA patients but in only 17% of heparin alone patients right-ventricular wall motion at 24 hours had improved from baseline and in 2% and 17%, respectively, it worsened (p = 0.005). rt-PA patients also had a significant decrease in right-ventricular end-diastolic area during the 24 hours after randomisation and a significant absolute improvement in pulmonary perfusion (14.6% vs 1.5%). No clinical episodes of recurrent PE were noted among rt-PA patients, but there were 2 fatal and 3 non-fatal clinically suspected recurrent PEs within 14 days in patients randomised to heparin alone. rt-PA rapidly improves right-ventricular function and pulmonary perfusion among patients with PE and may lead to a lower rate of adverse clinical outcomes.

Acute Disease↗
PubMed1993

A unique presentation of multicentric reticulohistiocytosis in pregnancy.

We describe a patient with multicentric reticulohistiocytosis who presented, during the second trimester of pregnancy, with symmetric polyarthritis, marked erythematous, pulsatile synovial swelling of the distal interphalangeal joints of both hands, and widespread telangiectasias. She did not have the typical skin manifestations of multicentric reticulohistiocytosis. The erythema and pulsatility of the synovial swellings of the distal interphalangeal joints resolved after delivery, but she continued to have widespread active synovitis, which did not resolve until treatment with low-dose oral methotrexate was instituted.

Adult↗
PubMed1993

Is aggressive heparinization necessary for elective PTCA?

The optimal level of heparin anticoagulation for elective PTCA is unknown. To determine if PTCA complications are related to the level of anticoagulation, serial ACT values were prospectively measured in 189 patients undergoing 201 elective PTCA procedures. The mean heparin dose before balloon inflation (pre-inflation) was 10,100 units, and the mean dose per procedure was 13,200 units. The mean pre-inflation ACT was 295 sec, but was < 300 sec in more than 50% of patients. Acute complications were not related to any ACT parameter and the development of new intracoronary thrombus was not observed. In elective PTCA procedures, the routine monitoring of ACT values is unnecessary when standard heparin doses are used.

Angioplasty, Balloon, Coronary↗
PubMed1993

Incorporation of oleic acid and eicosapentaenoic acid into glycerolipids of cultured normal human fibroblasts.

Confluent skin fibroblasts from normal humans were incubated in serum free medium with up to 100 nmole/mL eicosapentaenoic acid (EPA; bound to albumin in a 4.6:1 ratio) and compared with cells incubated with oleic acid (OA) at similar concentrations. The rate of [14C]OA incorporation into triacylglycerol (TG) (nmol/mg/h) was approximately 5-fold greater than that of [14C]EPA. The mass of TG formed after incubation of fibroblasts with EPA was also significantly lower than that formed with OA (43.2 +/- 9.3 vs. 59.5 +/- 6.6 micrograms/mg cell protein, respectively, P = 0.006). The addition of excess, unlabeled EPA reduced the rate of incorporation of [14C]OA into TG whereas unlabeled OA stimulated incorporation of [14C]EPA into TG. When the cells were preincubated with human serum basic proteins (BP I, II and III), the mass of TG formed (compared to baseline) was significantly higher with the basic proteins whether OA or EPA was studied. Only BP I significantly stimulated the mass of cell phospholipids, an effect that occurred with either OA or EPA in the medium. The results suggest that in cultured normal human fibroblasts, OA is a better substrate for TG synthesis than EPA and that this effect may be accentuated by the presence of the basis proteins.

Binding, Competitive↗
PubMed1993

Effect of gemfibrozil in men with primary isolated low high-density lipoprotein cholesterol: a randomized, double-blind, placebo-controlled, crossover study.

PURPOSE: To evaluate the efficacy of gemfibrozil in men with primary isolated low high-density lipoprotein cholesterol (HDL-C) levels. PATIENTS AND METHODS: Fourteen men with low levels of HDL-C but desirable total cholesterol levels received gemfibrozil in a randomized, double-blind, placebo-controlled, crossover trial. The men were placed on a National Cholesterol Education Program Step-Two Diet. They were randomly assigned to receive placebo and gemfibrozil each for 3 months, with a 1-month washout period between phases. RESULTS: Overall, gemfibrozil increased the total HDL-C concentration by 9.2% (p = 0.001), reduced triglyceride (TG) levels by 38% (p < 0.01), and significantly lowered the total cholesterol:HDL-C ratio (p = 0.01). Those with fasting TG levels of 1.07 mmol/L (95 mg/dL) or greater had a significant elevation in the HDL-C level (14.6%, p = 0.005) and a reduction in TG levels (50%, p = 0.002) with gemfibrozil; those with fasting TG levels less than 1.07 mmol/L had a smaller increase in the HDL-C level (4.1%, p > 0.05) and a smaller reduction in TG levels (15%, p = 0.02). There were no significant differences in the plasma levels of low density lipoprotein-cholesterol, HDL2-C, apolipoproteins (apo) A-I and B, or Lp(a). HDL3-C and apo A-II levels rose slightly. The adverse effects attributable to gemfibrozil were minimal. CONCLUSION: In men with desirable total cholesterol levels, gemfibrozil raises HDL-C and lowers TG levels to a similar extent as reported for hyperlipidemic men in the Helsinki Heart Study. These lipid-altering effects were most pronounced in those with the highest fasting TG levels.

Adult↗
PubMed1993

Erythrocyte membrane structure in bipolar affective disorder: a non-replication.

Fifteen bipolar patients were compared with sixteen controls in an attempt to replicate the findings of Pettegrew et al. in 1982 of decreased fluidity in the hydrocarbon core of the erythrocyte membrane. No significant differences were seen between groups. The present control group shows very similar membrane characteristics to the original control series; however, the bipolar patient group is not similar. Possible explanations are discussed.

Adult↗
PubMed1993

Glucocorticoid receptor binding to rat liver nuclei occurs without nuclear transport.

The binding of cell-free activated glucocorticoid receptor-steroid complexes from HTC cells to various preparations of HTC and rat liver nuclei has been examined under conditions that did or did not support the nuclear translocation of macromolecules via nuclear pores. To the best of our knowledge, this is the first such study with functionally active isolated nuclei. Conventionally prepared HTC nuclei were found to be porous, as determined from their inability to exclude the fluorescent macromolecule phycoerythrin (PE) at 4 degrees C. Thus the nuclear binding of activated complexes to these nuclei can not involve nuclear translocation. Further studies, using established conditions with sealed nuclei prepared from rat liver, revealed that nuclear translocation of PE containing a covalently linked, authentic nuclear translocation sequence could be obtained at 22 degrees C, but not at 4 degrees C. However, under the same conditions, activated glucocorticoid complexes displayed equal levels of nuclear binding at both temperatures. We therefore conclude that the current translocation conditions with intact rat liver nuclei are not sufficient to reproduce the nuclear transport of glucocorticoid complexes observed in intact cells. The nuclear binding that was seen with intact rat liver nuclei was not affected by aurintricarboxylic acid, which selectively inhibits protein-nucleic acid interactions. The antibody AP-64, shown to be specific for amino acids 506-514 of the nuclear translocation sequence of the rat glucocorticoid receptor, inhibited the nuclear binding of activated complexes, apparently by blocking receptor access to the nuclear membrane. Collectively, these data argue that activated complex binding to nuclei capable of nuclear translocation involves only an association with nuclear membrane components such as nuclear pores. Thus this system, and these reagents, may be useful in future studies of activated complex binding to nuclear pores.

Amino Acid Sequence↗
PubMed1993

Histologic determination of the ischemic threshold of muscle in the canine compartment syndrome model.

Our objective was to define the critical tissue pressure at which irreversible muscle damage occurs and to compare our results to those thresholds advocated in the orthopaedic literature. A standard plasma infusion compartment syndrome model was created in a canine model. Four dogs were in each of four experimental groups with compartment pressure maintained as follows: (a) 30 mm Hg with support of diastolic blood pressure to a level > 50 mm Hg; (b) 20 mm Hg less than diastolic pressure; (c) 10 mm Hg less than diastolic blood pressure; (d) a level equal to the animal's diastolic blood pressure. All animals were sacrificed 14 days after the procedure. Histology revealed the following: (a) tissues pressurized to 30 mm Hg in a normotensive dog demonstrated no significant abnormalities; (b) tissues pressurized to 20 mm Hg less than diastolic revealed occasional cells undergoing regeneration but no evidence of infarction or fibrosis; (c) tissues pressurized to 10 mm Hg less than diastolic showed scattered small areas of infarction and fibrosis; and (d) tissues pressurized to diastolic blood pressure demonstrated more widespread infarction and scarring. The ischemic threshold of muscle, beyond which irreversible tissue damage occurs, is directly related to the difference in compartment and perfusion pressure. Our findings document this pressure to be 10 mm Hg less than diastolic blood pressure or within 30 mm Hg of mean arterial pressure. This data refutes the use of absolute tissue pressure values as a guide to the necessity of fasciotomy. To abort an impending compartment syndrome and avoid irreversible tissue injury and their sequelae, fasciotomy should be done if tissue pressure reaches within 10-20 mm Hg of diastolic pressure.

Animals↗
PubMed1993

Identification of alcoholism and depression in a geriatric medicine outpatient clinic.

OBJECTIVE: To examine the utility of brief screening instruments for alcoholism and depression in the frail elderly medical outpatient. DESIGN: Cross-sectional examination. SETTING: Coffey Geriatric Outpatient Clinic, The Mount Sinai School of Medicine, New York City. SUBJECTS: Convenience sample of 84 consenting male and female outpatients over 62 years of age. MEASUREMENTS: The Michigan Alcohol Screening Test (MAST), CAGE questionnaire, and the Geriatric Depression Scale (GDS). RESULTS: Four subjects (5%) scored positive for alcoholism on the MAST, all of whom had been previously identified by the geriatric clinic staff. In contrast, the CAGE only identified 1 (1.4%) alcoholic subject. Thirty-two percent of subjects scored positive for depression on the GDS, yet only one-third of these depressed patients had been previously identified by the geriatric team as depressed. Among the depressed outpatients, 19% were labeled anxious and received anxiolytics from the geriatric medical staff. CONCLUSIONS: Whereas brief alcohol screening instruments to detect alcoholism did not augment the geriatrician's clinical assessment, screening for depression with the GDS may be an important adjunct to the identification of depression in geriatric medical patients.

Aged↗
PubMed1993

Effect of dietary lipid peroxides on metabolism of serum chylomicrons in rats.

We have found that in rats the peroxide content of chylomicrons (CM) is determined by the lipid peroxide concentration in the diet, indicating that dietary lipid peroxides are incorporated into the lymph CM. Moreover, these incorporated lipid peroxides influence the normal CM metabolism. When radiolabeled CM were injected into rats, there was no difference in the initial plasma removal between CM prepared from oil with low peroxide content (control CM) and CM prepared from oil with high peroxide content (oxidized CM). However, the tissue distribution of the labels indicated that the hepatic uptake of CM decreased with increasing lipid peroxide content of CM. At 10 min after injection of CM, liver uptake of cholesterol label was 48.39 +/- 3.08% for control CM and 31.41 +/- 10.73% for oxidized CM. Vitamin E enrichment of control CM increased their hepatic uptake to 61.07 +/- 0.83%. Additionally, binding of oxidized CM to the heart endothelium increased from 2.55 to 3.60% compared with binding of control CM. When the hydrolysis of control and oxidized CM by endothelial lipoprotein lipase (LPL) was tested in a heart perfusion system, we found that after 30 min, 56.51 +/- 5.81% of control and 76.82 +/- 1.75% of oxidized CM were not hydrolyzed and remained in the perfusate. Thus our results indicate that the altered metabolism of oxidized CM may be related to a reduced hydrolysis rate by endothelial LPL.

Animals↗
PubMed1993

Increased vasopressin secretion and release in mice transgenic for the rat arginine vasopressin gene.

The rat arginine vasopressin (AVP) genomic sequence has been utilized to develop a line of transgenic mice homozygous and heterozygous for the transgene. Expression of the rat AVP gene was demonstrated by Southern blotting and resulted in increased amounts of AVP in hypothalamus and frontotemporal brain cortex. Secretion of AVP from the neurohypophysial system results in an increased concentration of the hormone in the plasma and in an increased excretion in the urine in amounts three to five times those of normal mice. Extraneural ectopic hormone production was found only in the pancreas. Despite chronic hypersecretion of AVP, 24-hour urine volume and osmolality did not show evidence of increased antidiuretic hormone action on the kidney, so that, under basal conditions, the water balance in the animals is unaffected.

Animals↗
PubMed1993

Induction of hepatic bilirubin and drug metabolizing enzymes by individual herbs present in the traditional Chinese medicine, yin zhi huang.

Yin Zhi Huang (YZH) is a decoction of four plants which is widely used in Asia to treat neonatal jaundice. This study compares the ability of phenobarbital and the individual herbs comprising YZH, Artemisia, Gardenia, Rheum, and Scutellaria baicalensis, to induce hepatic drug and bilirubin metabolizing enzymes in rats. Herbal decoctions (30 ml/kg/day) or phenobarbital (60 mg/kg/day) were administered for 5 days. Only phenobarbital increased cytochrome P-450 levels whereas Gardenia slightly decreased levels. Artemisia, Rheum and phenobarbital increased bilirubin glucuronyl transferase activity. Glucuronidation of alpha-naphthol was increased by Gardenia and phenobarbital, whereas Artemisia and Rheum were ineffective inducers. Phenobarbital was the most effective inducer of glutathione-S-transferase (GSHT) activity. Phenobarbital and Gardenia both induced delta 5-3-ketosteroid isomerase activity, a marker for the Ya subunit of GSHT responsible for intracellular bilirubin transport in liver. The selective patterns of enzyme induction suggest potential value for using specific plant decoctions to modify drug and bilirubin metabolic pathways.

Animals↗
PubMed1993

Decreased postprandial response to a fat meal in normotriglyceridemic men with hypoalphalipoproteinemia.

The plasma level of high density lipoprotein cholesterol (HDL-C) has been reported to be inversely correlated with the level of triglycerides (TGs) and the magnitude of postprandial lipemia because subjects with low HDL-C accompanying high TG levels often have an increased postprandial response to a fat load. However, information is limited regarding the postprandial response to a fat load in subjects with low HDL-C and normal fasting TG values (hypoalphalipoproteinemia [hypoalpha]). We administered an oral fat load (70 g/m2 of body surface area) to six subjects with hypoalpha and six aged-matched control subjects. Plasma levels of lipids and lipoproteins and the mass of triglyceride-rich lipoproteins (TRLs: Sf > 100 and Sf 20-100) were measured every 2 hours for 8 hours. The mass and chemical composition of Sf > 100 or Sf 20-100 TRLs were not different between the fasting groups; postprandial Sf > 100 but not Sf 20-100 TRLs (mean +/- SD) was significantly lower in subjects with hypoalpha (200.4 +/- 64.8 mg/dL versus 110.6 +/- 50.9 mg/dL; analysis of variance, F test, p = 0.04). In the hypoalpha subjects, the compositions of postprandial Sf > 100 TRLs were TG poor and cholesterol and phospholipid enriched (p < 0.001) while the Sf 20-100 TRLs were enriched in cholesterol and phospholipid but relatively protein depleted (p = 0.002).(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1993

Transgenic mouse models of vasopressin expression.

Arginine vasopressin is a nine-amino acid neuropeptide hormone important in the regulation of water metabolism. It also may have a role in other physiological functions, such as blood pressure regulation and the response to stress. Whole animal studies have provided a good understanding of vasopressin physiology and regulation of the normal vasopressin gene, and in vitro cell culture studies have demonstrated important features of the intracellular regulation of vasopressin gene expression. Transgenic mice provide useful models for the study of the in vivo regulation of gene expression. Previously reported mouse lines transgenic with vasopressin gene constructs have not expressed the transgene in a tissue distribution similar to that detected for the endogenous mouse vasopressin gene. An 8.2-kb genomic construct of the rat vasopressin gene, including 3 kb each of 5' and 3' flanking sequences, has been used to develop a line of transgenic mice. These animals express the transgene in a tissue-specific manner, demonstrate appropriate osmotic regulation of transgenic vasopressin mRNA, and have normal water metabolism. Animals homozygous for the 8.2-kb transgene have increased basal plasma levels of vasopressin peptide but have no apparent change in basal water metabolism. The findings with this and other previously reported mouse lines transgenic for vasopressin constructs provide a basis for developing future transgenic lines to study the in vivo regulation of the vasopressin gene.

Animals↗
PubMed1993

Expression of the rat arginine vasopressin gene in transgenic mice.

A line of mice has been developed which are transgenic for an 8.2-kilobase (kb) genomic clone of the rat vasopressin (VP) gene. Using a polymerase chain reaction technique, the rat VP (rVP) transgene was shown to have tissue-specific mRNA expression in the hypothalamus, temporal lobe, parietal cerebral cortex, cerebellum, and posterior pituitary, similar to the tissue distribution of endogenous mouse and rat VP expression. Expression of transgenic rVP mRNA was also found in the lung and pancreas of the transgenic mice, sites of known ectopic expression of VP. Using two methods, Northern blot analysis with species-specific cRNA probes and a quantitative polymerase chain reaction technique, the quantity of rVP transgene mRNA was shown to regulate appropriately in response to an osmotic stimulus. After 72 h of water deprivation, the quantity of transgenic rVP mRNA increased 6.8 +/- 3.0-fold. This was not significantly different than the fold increase in mouse VP mRNA quantity seen in nontransgenic mice (4.8 +/- 1.5) but was significantly different (P < 0.05) than the 1.2 +/- 0.03-fold increase in rat VP mRNA seen in normal rats after water deprivation. In the rat hypothalamus, VP mRNA poly(A) tail length increases with osmotic stimulation, while in the mouse it does not. The poly(A) tail of transgenic rVP mRNA expressed in mouse hypothalamus did not increase in length after osmotic stimulation.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗