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Biomedical subjects

M A Miller

Publications and source records attributed to M A Miller.

At least 253 records · Page 14Linked to original sources
PubMed1994

Fetal hemoglobin and potassium in isolated transferrin receptor-positive dense sickle reticulocytes.

A subset of sickle cells have an increased density at the reticulocyte stage of development, indicating that they are either abnormally dense upon release from the bone marrow or become dense quickly in the circulation. These cells are of interest because they most likely have severely disrupted cation regulation and a short lifespan. Based on the distribution of fetal hemoglobin (HbF) in the density fractions of sickle red blood cells (RBCs) and in vitro studies of cellular K+ loss, it seems likely that HbF content is an important in vivo determinant of dense cell formation. In this study, we tested the hypothesis that young, dense cells have low HbF content. Sickle RBCs were first separated into light and dense fractions. Reticulocytes were isolated from unfractionated cells and from each density fraction with an immunomagnetic technique directed against transferrin receptors (TfR) and assayed for the percentage of HbF and K+/Hb ratio. TfR+ reticulocytes isolated from unfractionated cells had a much lower HbF content when compared with all the unfractionated RBCs. This is most likely caused by enrichment of F cells because of a longer circulation life span. Heavy TfR+ reticulocytes had a K+/Hb ratio similar to that measured in the entire dense population and contained very low levels of HbF, averaging 2.5% of the level in all RBCs, 11.7% of the level in all TfR+ reticulocytes, and 4.0% of the level in all dense RBCs. These findings suggest that TfR+ dense cells derive predominantly from non-F cells. Furthermore, the amount of HbF in the circulating dense cells suggests that many of these cells do not derive from the TfR+ dense cells.

Anemia, Sickle Cell↗
PubMed1994

Role of methionine 230 in intramolecular electron transfer between the oxyferryl heme and tryptophan 191 in cytochrome c peroxidase compound II.

The kinetics of electron transfer from cytochrome c (CC) to yeast cytochrome c peroxidase (CcP) compound I were studied by flash photolysis and stopped-flow spectroscopy. Flash photolysis studies employed horse CC derivatives labeled at specific lysine amino groups with (dicarboxybipyridine)bis-(bipyridine)ruthenium (Ru-CC). Initial electron transfer from Ru-CC reduced the indole radical on Trp-191 of CcP compound I [CMPI(IV,R.)], producing CMPII(IV,R). This reaction was biphasic for each of several Ru-CC derivatives, with rate constants which varied according to the position of the Ru label. For Ru-27-CC labeled at lysine 27, rate constants of 43,000 and 1600 s-1 were observed at pH 5.0 in 2 mM acetate. After reduction of the indole radical by Ru-CC, intramolecular electron transfer from Trp-191 to the oxyferryl heme in CMPII(IV,R) was observed, producing CMPII(III,R.). The rate constant and extent of this intramolecular electron transfer reaction were independent of both the protein concentration and the Ru-CC derivative employed. The rate constant decreased from 1100 s-1 at pH 5 to 550 s-1 at pH 6, while the extent of conversion of CMPII(IV,R) to CMPII(III,R.) decreased from 56% at pH 5 to 29% at pH 6. The reaction was not detected at pH 7.0 and above. The pH dependence of the rate and extent of this internal electron transfer reaction paralleled the pH dependence of the rate of bimolecular reduction of CMPII(IV,R) by native horse CC measured by stopped-flow spectroscopy at high ionic strength.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1994

Interaction domain for the reaction of cytochrome c with the radical and the oxyferryl heme in cytochrome c peroxidase compound I.

Site-directed mutants of cytochrome c peroxidase (CcP) were created to modify the interaction domain between CcP and yeast iso-1-cytochrome c (yCC) seen in the crystal structure of the CcP-yCC complex [Pelletier & Kraut (1992) Science 258, 1748-1755]. In the crystalline CcP-yCC complex, two acidic regions of CcP contact lysine residues on yCC. Mutants E32Q, D34N, E35Q, E290N, and E291Q were used to examine the effect of converting individual carboxylate side chains in the acidic regions to amides. The A193F mutant was used to test the effect of introducing a phenyl moiety at the point of closest contact between CcP and yCC in the crystal structure. Stopped-flow experiments carried out in 310 mM ionic strength buffer at pH 7 revealed that yCC initially reduced the indole radical on Trp-191 of the parent CcP compound I with a bimolecular rate constant ka = 2.5 x 10(8) M-1 s-1. A second molecule of yCC subsequently reduced the oxyferryl heme of compound II with a rate constant kb = 5 x 10(7) M-1 s-1. The bimolecular rate constants ka and kb were affected in parallel by each mutation examined. CcP mutants D34N and E290N that are closest to a complementary yCC lysine residue in the crystalline CcP-yCC complex gave the lowest values for ka and kb, which were 25-50% of the values of the CcP parent. Mutants E32Q and E291Q that are removed from the interaction domain gave the same ka and kb values as the CcP parent.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1994

Fatal cerebral coenurosis in a cat.

A 6-year-old cat that was laterally recumbent and panting was evaluated because of a 10-day history of progressive neurologic abnormalities. Despite aggressive treatment, the cat died on the day of admission. At necropsy, a 1.5-cm-diameter, fluid-filled cyst was found in the white matter of the left cerebrum. The cyst was identified as a coenurus of Taenia serialis, on the basis of the cyst wall, distribution of scolices, and the shape and dimensions of rostellar hooks. Scolices were found in varying stages of ontogeny, ranging from undifferentiated to nearly mature. Taenia serialis has a canid-lagomorph life cycle, with cats being accidental intermediate hosts. However, the potential exists for rare zoonotic transmission and subsequent serious disease in human beings and other accidental intermediate hosts.

Animals↗
PubMed1994

Amaranthus retroflexus (redroot pigweed) poisoning in cattle.

Amaranthus retroflexus (redroot pigweed)-induced nephrotoxicity was diagnosed in 6 herds of cattle from 3 counties in southwest Missouri. Forty-eight cows and calves died and another 35 were clinically affected. Serum urea nitrogen concentration, determined in 4 affected calves, was between 55 and 284 mg/dl, and serum creatinine concentration was between 6.7 and 29.9 mg/dl. Postmortem examination of affected cows and calves revealed amber-colored fluid in peritoneal cavities and retroperitoneal perirenal edema. Histologic examination of kidney sections revealed widespread degeneration and necrosis of proximal and distal tubules. Compared with archived kidney sections from cattle with Quercus (oak) poisoning, distal renal tubules were more severely affected. Oak poisoning also was associated with a higher prevalence of interstitial fibrosis and renal tubular oxalosis. We concluded that ingestion of the aerial and leafy portions of pigweed by cattle in drought situations does not necessarily lead to nitrate-induced sudden death associated with consumption of the nitrate-containing stems.

Animals↗
PubMed1994

Reaction of horse cytochrome c with the radical and the oxyferryl heme in cytochrome c peroxidase compound I.

The reactions of recombinant cytochrome c peroxidase [CcP(MI)] and a number of CcP(MI) mutants with native and ruthenium-labeled horse ferrocytochrome c have been studied by stopped-flow spectroscopy and laser flash photolysis. At 100 mM ionic strength, pH 7.5, native horse ferrocytochrome c reduces the radical on the indole group of Trp-191 in cytochrome c peroxidase compound I (CMPI) with a second-order rate constant of 1.3 x 10(8) M-1 s-1. Ferrocytochrome c then reduces the oxyferryl heme Fe(IV) in CMPII with a rate constant of 2.0 x 10(6) M-1 s-1. The rate constant for the reduction of the radical is nearly independent of pH from 5 to 8, but the rate constant for reduction of the oxyferryl heme Fe(IV) increases 33-fold as the pH is decreased from 8 to 5. This increase in rate is correlated with the pH dependence of the electron transfer equilibrium between the radical and the oxyferryl heme Fe(IV) in the transient form of CMPII. The second-order rate constants for reduction of the radical and the oxyferryl heme in the mutants Y39F, Y42F, H181G, W223F, and Y229F are nearly the same as for wild-type CcP(MI). The intracomplex rate constants for reduction of the radical in these mutants by the ruthenium-labeled cytochrome c derivatives are also similar to that for CcP(MI). This rules out a direct role for these aromatic residues in electron transfer.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1994

Quinapril hydrochloride effects on renal function in patients with renal dysfunction and hypertension: a drug-withdrawal study.

Patients with mild to moderate hypertension (diastolic blood pressure > or = 95 and < or = 115 mmHg) and renal dysfunction entered one of two studies to assess the safety of efficacious daily doses of quinapril on renal function and blood pressure. Twenty-four patients with moderate renal impairment (MRI) (creatinine clearance > 30 and < or = 60 ml/min) entered 24 weeks of open-label quinapril treatment; 31 patients with chronic renal failure (CRF) (creatinine clearance < 30 ml/min) entered 16 weeks of open-label quinapril treatment. Patients with MRI initially received quinapril 5 mg once daily (qd) followed by titration to a maximum dosage of 40 mg/day (furosemide optional at 40 mg only). Patients with CRF initially received quinapril 2.5 mg qd and were titrated up to 20 mg/day (furosemide optional). Open-label quinapril treatment resulted in significant decreases in mean systolic (SBP) and diastolic (DBP) blood pressure. The 20 patients with MRI and the 28 with CRF who completed the open-label phase were then randomly assigned to continue active drug or to receive placebo in a 4-week, double-blind, drug-withdrawal phase. During the double-blind withdrawal phase, placebo-treated patients had significant increases in mean SBP and DBP from the end of open label. Creatinine clearance was essentially unchanged following open-label quinapril treatment or quinapril withdrawal. In conclusion, in patients with mild to moderate hypertension and renal dysfunction, quinapril in dosages of 5-40 mg qd for patients with MRI and 2.5 to 20 mg qd for patients with CRF significantly reduces blood pressure without adversely affecting renal function.

Adult↗
PubMed1994

The molecular biology of esophageal and gastric cancer and their precursors: oncogenes, tumor suppressor genes, and growth factors.

The evolution of sequential histological changes from normal cells through invasive cancer affords the cancer biologist the opportunity to identify separate molecular steps involved in cancer progression. As one studies the development of human carcinoma, it becomes apparent that multiple genetic alterations affecting both cellular proto-oncogenes and tumor suppressor genes are involved during the development and progression of both esophageal and gastric cancers. The different histological forms of both esophageal and gastric carcinomas as well as their differing etiologies result in the possibility that a spectrum of genetic changes is involved in different tumor types. p53 abnormalities occur frequently in tumors arising in both organs, and in both sites p53 abnormalities can be observed in precancerous lesions as well as in overt cancer. Subsequent abnormalities affecting other genes (eg, epithelial growth factor receptors [EGFRs]) potentially enhance the growth potential of tumors. This review focuses on abnormalities of oncogenes, tumor suppressor genes, and growth factors commonly found in cancers of the esophagus and stomach.

Carcinoma, Squamous Cell↗
PubMed1994

Coexpression of neurotensin and c-fos mRNAs in rat neostriatal neurons following acute haloperidol.

Our previous studies have shown several fold induction of neurotensin/neuromedin N (NT/N) primary transcripts and mature cytosolic mRNA in the dorsolateral region of the rat neostriatum (DLSt) following a single dose of the neuroleptic, haloperidol. The apparent enhancement of NT/N gene transcription by haloperidol is preceded by increases in the expression of c-fos mRNA in the same region. The present study used double-labeling in situ hybridization technique to study cellular localization of NT/N and c-fos mRNA following acute haloperidol treatment. Simultaneous detection of NT mRNA and c-fos mRNA was achieved using cRNA probes synthesized in vitro with digoxigenin- and 35S-labeled UTP, respectively. Interestingly, approximately 75% of DLSt neurons expressing NT/N mRNA also displayed c-fos mRNA in rats treated with haloperidol (1 mg/kg, i.p.) for 1 h. Colocalization of c-fos mRNA in haloperidol-responsive NT neurons in the DLSt suggests that haloperidol's induction of NT/N gene transcription may involve participation of the transcription factor Fos and the AP-1 consensus sequence in regulatory region of the NT/N gene.

Animals↗
PubMed1994

Activation of galanin pathways across puberty in the male rat: galanin gene expression in the bed nucleus of the stria terminalis and medial amygdala.

Galanin and vasopressin are coexpressed in the bed nucleus of the stria terminalis and medial amygdala of the male rat. In adult males, the level of gene expression for both peptides in these regions is dependent on circulating levels of testosterone. We hypothesized that galanin messenger RNA levels would be enhanced in adult males compared with prepubertal males due to the rise in plasma testosterone levels. We used in situ hybridization and quantitative autoradiography to measure galanin messenger RNA in cells of the bed nucleus of the stria terminalis and medial amygdala of prepubertal and adult male rats. Our results show that significantly (P < or = 0.05) more galanin messenger RNA expressing neurons are detectable in the bed nucleus of the stria terminalis of adult compared with prepubertal male rats. In contrast, no differences were observed between the groups in the number of labeled neurons detected within the medial amygdala. However, the average labeling intensity was significantly enhanced in both the bed nucleus of the stria terminalis (P < or = 0.001) and medial amygdala (P < or = 0.001) of adult compared with prepubertal animals. The present findings are consistent with the hypothesis that gonadal hormones regulate galanin gene expression in some brain regions and suggest that the activation of the hypothalamic-pituitary-gonadal axis which occurs naturally with puberty is associated with activation of galanin pathways in the bed nucleus of the stria terminalis and medial amygdala.

Amygdala↗
PubMed1994

Activation of galanin pathways across puberty in the male rat: assessment of regional densities of galanin binding sites.

Galanin-like immunoreactivity and galanin messenger RNA levels increase across puberty in neurons of gonadal steroid-dependent brain nuclei. We hypothesized that this activation and the associated increase in endogenous galanin release would result in changes across puberty in both galanin binding density and the level of receptor occupancy. Here we have assessed the density of galanin binding sites in several brain regions of prepubertal and adult male rats with or without GTP to induce dissociation of endogenous galanin from its binding sites. The developmental changes in the level of receptor occupancy were used as an indirect measure of changes in neuropeptide release from galanin expressing neurons. In standard binding conditions (buffer preincubation), 125I-labeled galanin binding showed a generalized decline in adult brains (34-68%) compared with prepubertal levels in most regions of the telencephalon and diencephalon. Following preincubation with 10(-5) M GTP, galanin binding showed a dramatic increase in most regions of the adult (152-504%) and several regions of the prepubertal brain (132-245%) over their standard binding levels. However, this increase was greatest in adult animals. Finally, although preincubation of brain slices with GTP eliminated most of the apparent age-related differences observed in standard binding conditions, several brain regions of the adult brain continued to show a significant reduction (38-76%) in 125I-labeled galanin binding compared with prepubertal animals. Only one region, the lateral preoptic area, exhibited enhanced 125I-labeled galanin binding in adult (160%) compared with prepubertal brain after GTP preincubation.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1994

2.2 A structure of oxy-peroxidase as a model for the transient enzyme: peroxide complex.

The Fe+3-OOH complex of peroxidases has a very short half life, and its structure cannot be determined by conventional methods. The Fe+2-O2 complex provides a useful structural model for this intermediate, as it differs by only one electron and one proton from the transient Fe+3-OOH complex. We therefore determined the crystal structure of the Fe+2-O2 complex formed by a yeast cytochrome c peroxidase mutant with Trp 191 replaced by Phe. The refined structure shows that dioxygen can form a hydrogen bond with the conserved distal His residue, but not with the conserved distal Arg residue. When the transient Fe+3-OOH complex is modelled in a similar orientation, the active site of peroxidase appears to be optimized for catalysing proton transfer between the vicinal oxygen atoms of the peroxy-anion.

Binding Sites↗
PubMed1994

Clinical nurse specialist practice: facilitators and barriers.

Responding to a survey containing open-ended questions, 288 Master's-prepared CNSs across the United States identified role components that are a part of their practice, described health care system changes affecting implementation of their roles, and described resources that were or would be supportive of successful role implementation. Role categories related to patient care/consultation, education, research, administration, and professional development were ranked in that order according to time spent in each activity. Issues related to institutional cost containment and employment of CNSs were described most frequently as affecting role implementation. Nursing administrative support was indicated as the resource most crucial for role success. Specific suggestions for successful implementation of CNS role are described.

Consultants↗
PubMed1994

Adenylate and guanylate cyclase activity in the penis and aorta of the diabetic rat: an in vitro study.

OBJECTIVE: To investigate the role of adenylate and guanylate cyclases in the mediation of erection in diabetic rats. MATERIALS AND METHODS: Hyperglycaemic diabetes mellitus was induced in 35 rats using streptozotocin. Two months later the penises and aortae of these rats were excised and cut into rings or segments before being treated with varying concentrations of acetylcholine (Ach), sodium nitroprusside (NaNP), prostaglandin E1 (PGE1) and adrenaline (AD). The levels of adenosine 3'5'-cyclic monophosphate (cAMP) and guanosine 3'5'-cyclic monophosphate (cGMP) so generated were measured by radioimmunoassay and the results compared with those from seven age-matched control rats that had not been given streptozotocin. RESULTS: Ach-stimulated cGMP synthesis was impaired in the aortae in diabetic rats. Ach-stimulated cGMP synthesis was undetectable in the penis. NaNP-stimulated cGMP and PGE1-stimulated cAMP synthesis was enhanced in both the penises and aortae in diabetic rats compared with controls. AD-stimulated cAMP synthesis was enhanced in the aorta in diabetic rats compared with controls, but AD had no effect on cAMP synthesis in the penis. CONCLUSION: Ach-stimulated nitric oxide (NO) synthesis is impaired in the vasculature of diabetic rats and an Ach-NO axis may not be present in the penis of the rat. The enhanced capacity of the penis and vasculature to generate cAMP and cGMP may constitute an adaptive response to counteract the reduction in receptor-linked NO release. Impaired adenylate or guanylate cyclase activity does not contribute to erectile dysfunction in diabetic rats.

Adenylyl Cyclases↗