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M R Miller

Publications and source records attributed to M R Miller.

161 records · Page 9Linked to original sources
PubMed1976

Reversible inactivation of phenylalanine hydroxylase by catecholamines in cultured hepatoma cells.

Phenylalanine hydroxylase in Reuber H4 hepatoma cell cultures can be rapidly inactivated by the addition of epinephrine, norepinephrine, dopamine, or 3,4-dihydroxyphenylalanine, in order of decreasing effectiveness, to the culture medium. The enzyme was 50% inactivated in 1 hour by 25 muM (R)-epinephrine or 45 muM (R)-norepinephrine in the medium. High concentrations of epinephrine caused a 70% inactivation in 15 min. Phenylalanine hydroxylase appears to be reversibly inactivated by epinephrine within the cells; since washing the compound off the cell cultures resulted in a rapid restoration of enzyme activity (40% in 1 hour), cycloheximide had little effect on the initial rate of recovery of enzyme activity and the same amount of phenylalanine hydroxylase antigen per cell was isolated from treated and normal cultures. Both (S)- and (R)-epinephrine inactivated the enzyme, and 0.1 mM desmethylimipramine, an inhibitor of amine transport, significantly decreased the effect of epinephrine on the hydroxylase activity. The possibility, suggested by the above results, that epinephrine might be directly inactivating phenylalanine hydroxylase within the cells was supported by the finding that purified rat liver phenylalanine hydroxylase would be 50% inactivated by 1.5 muM epinephrine in 10 min.

Carcinoma, Hepatocellular↗
PubMed1976

Mechanism of inactivation of phenylalanine hydroxylase by p-chlorophenylalanine in hepatome cells in culture. Two possible models.

The mechanism by which p-chlorophenylalanine specifically reduces phenylalanine hydroxylase activity in rat liver in vivo and in Reuber H4 hepatoma cells in culture has been investigated. Chromatography on hydroxylapatite of liver extract from rats injected with p-chlorophenylalanine showed that the compound differentially affected the three normal phenylalanine hydroxylase isoenzymes (I, II, and III); isoenzymes II and III were completely absent after the treatment, but isoenzyme I was only reduced in quantity compared with normal adult rats. Normal Reuber H4 cells only possess isoenzyme I; treatment with p-chlorophenylalanine yielded a reduced level of enzyme activity which appeared to be noraml isoenzyme I by both chromatographic and kinetic criteria. There is evidence, based on immunochemical techniques, that cultures grown in the presence of p-chlorophenylalanine have significantly reduced levels of phenylalanine hydroxylase antigen, and that p-chlorophenylalanine inactivates phenylalanine hydroxylase at or near the time of enzyme synthesis. The bulk of enzyme synthesized prior to the addition of the compound appears unaffected by it. There is no indication that protein synthesis itself is affected by p-chlorophenylalanine. In addition, p-chlorophenylacetate was found to inactivate phenylalanine hydroxylase in an apparently identical manner with p-chlorophenylalanine, which almost certainly eliminates from consideration any mechanism of inactivation specifically requiring an amino acid. Finally, effects of cycloheximide and chlorophenylalanine were compared. Taken together, the data lead to two possible models for the inactivation of the enzyme. The model most consistent with all data requires (predicts) the existence of a proenzyme form of phenylalanine hydroxylase which can be specifically inactivated by p-chlorophenylalanine.

Animals↗
PubMed1975

p-Chlorphenylalanine effect on phenylalanine hydroxylase in hepatoma cells in culture.

We have investigated the p-chlorophenylalanine-dependent loss of phenylalanine hydroxylase activity in cultured hepatoma cells. The similarity of the effect of p-chlorophenylalanine on phenylalanine hydroxylase in the hepatoma cells and that reported from studies in vivo indicates that the loss of phenylalanine hydroxylase activity is due to a direct interaction of the amino acid analogue with the liver. We can find no evidence that the loss of phenylalanine hydroxylase activity is due to: a direct inactivation of the hydroxylase by p-chlorophenylalanine or an inhibitor produced by p-chlorophenylalanine treatment; an effect similar to that of p-fluorophenylalanine; or leakage of enzyme from the cells during p-chlorophenylalanine treatment. The data presented indicate: (a) the p-chlorophenylalanine effect is rather specific for phenylalanine hydroxylase; (b) following p-chlorophenylalanine removal, new protein synthesis is necessary for restoration of the hydroxylase activity; (c) the rate of loss of phenylalanine hydroxylase activity after the addition of p-chlorophenylalanine is much faster than the rate of restoration of the hydroxylase activity after removal of p-chlorophenylalanine; (d) even in the presence of p-chlorophenylalanine, hydrocortisone greatly stimulates the hydroxylase activity; (e) the cell density-dependent increase of phenylalanine hydroxylase activity is blocked by p-chlorophenylalanine. A discussion of the possible mechanisms of p-chlorophenylalanine-dependent loss of phenylalanine hydroxylase is presented. To measure very low leanine-dependent loss of phenylalanine hydroxylase is presented. To measure very low levels of phenylalanine hydroxylase activity, a new procedure, based on isotope dilution, was developed for isolating the tyrosine formed during the enzymatic reaction.

Animals↗
PubMed1975

The cochlear nuclei of lizards.

The cochlear nuclei of 14 lizard species (eight families) were studied in normal animals and in a small series of animals with lesions of the posterior division of the eight nerve. The development of the cochlear nuclei is division of the eighth nerve. The development of the cochlear nuclei is directly related to the length and complexity of the papilla basilaris. The best development of basilar papillae and cochlear nuclei is found in teiid and gekkonid lezards, and an intermediate grade of development in scincid, lacertid, and anguid lizards. A lesser degree of development occurs in the iguanids, and practically no cochlear nuclei differentiation is observed in Chameleo. Two well defined cochlear nuclei are found in most lizard families: nucleus angularis (NA) AND nucleus magnocellularis medialis (NMM). NA is located in the cephalic third of the acoustic tubercle and contains variably sized darkly staining cells. NMM is the most caudally located nuclear group and is characterized by regularly round to ovoid cells. The development of NMM is more closely related to papilla basilaaris length and complexity than is NA. Two less well defined cochlear nuclei are also thought to be present. In the region beteen NA and NMM is a sparsely cell-populated area, nucleus magno-cellularis lateralis (NML), which consits of a variety of small, darkly staining cells and large, pale staining cells which are usually laterally located in the nucleus. Like NMM, Nml is better developed in lizards ith more complex papillae basilares. Nucleus laminaris may be represented by a few fusiform cells in the ventral portion of the NML region.

Animals↗
PubMedDate not supplied

Antibody-mediated red blood cell agglutination resulting in spontaneous echocardiographic contrast.

Spontaneous echocardiographic contrast is well reported in states of low flow and low shear stress, and the primary blood component involved has been reported as red blood cells via rouleaux formation. This report describes the occurrence of spontaneous echocardiographic contrast from a unique mechanism of IgM-mediated red blood cell agglutination and describes the clinical sequelae.

Anemia, Hemolytic, Autoimmune↗
PubMedDate not supplied

A five-generation family with late-onset progressive hereditary hearing impairment due to cochleosaccular degeneration.

Cochleosaccular dysplasia or degeneration (Scheibe degeneration) is considered the most common cause of profound congenital hearing impairment, and accounts for approximately 70% of cases 2 with hereditary deafness. A five-generation family with hereditary hearing impairment associated with cochleosaccular degeneration has recently been identified. The diagnosis of classical Scheibe degeneration was based on histopathological findings in the temporal bones of the proband, a 61-year-old profoundly deaf male. Auditory structures in the brainstem of the proband were also studied. Twenty-two members of the family were contacted for surveys and blood samples. Of these, 6 males and 2 females have hearing impairment. Complete audiological evaluation was done on 12 family members, and prior audiologic records of the proband and affected family members were available for study. Affected family members suffer a mild bilateral high-frequency hearing loss during childhood and adolescence, and progress to moderate-to-profound deafness in the second and third decades of life. The family is suitable for linkage analysis and does not map to previously reported loci harboring autosomal dominant, nonsyndromic hereditary hearing impairment genes. The genetic study of this family will be helpful in identifying the genes which, when mutated, result in Scheibe degeneration.

Acoustic Impedance Tests↗
PubMedDate not supplied

Methodologic challenges in developing and implementing measures of quality for child health care.

OBJECTIVE: To review the major building blocks in measurement of quality for child health care, with recommendations for future research. METHODS: We describe a framework of building blocks for quality measurement and discuss how an investigator's choices for each component are constrained by the special features of child health care. RESULTS: Methodologic challenges for children's health care include developmental change and dependency on others, fragmentary care and inadequate health care data, unusual care settings, potential for long-term consequences, proxy reporting of outcomes and patient experience, small sample sizes, and lack of evidence that links processes and outcomes of care and of methods for risk adjustment. We cite examples of child-specific measures of quality that illustrate solutions to these challenges. CONCLUSIONS: Children are different from adults, and measures of health care quality for children must differ from those for adults. We suggest future research on measures of quality directed toward overcoming the methodologic problems specific to child health care.

Adolescent↗