Estimating the association of phosphodiesterase with calmodulin in intact cells.
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3',5'-Cyclic-AMP Phosphodiesterases↗
Biomedical subjects
J R Miller
Publications and source records attributed to J R Miller.
Ventilator use in progressive neuromuscular disease: impact on patients and their families.
The question of extending the lives of patients with progressive neuromuscular disease by means of ventilator support is controversial. It has been documented that ventilators can prolong the lives of patients with Duchenne muscular dystrophy by between two and 25 years, but few studies have assessed the effects of their use on individual or family functioning, or the quality of survival. A sample of patients with progressive disease and families living in the community was surveyed to assess these effects. Most of the sample were satisfied with their decision to extend life with the ventilator, but they found the experience significantly stressful in the home. Both patients and families believed their health-care team could have met their needs more comprehensively. Recommendations are made for health-care professionals working with patients with late-stage neuromuscular disease who need to use ventilators.
Adolescent↗
Isolation and expression of a new mouse homeobox gene.
A homeobox-containing clone has been isolated from an adult mouse kidney cDNA library and shown by DNA sequence analysis to be a new isolate, Hox-6.1. A genomic clone containing Hox-6.1 has been isolated and found to contain another putative homeobox sequence (Hox-6.2), within 7 kb of Hox-6.1. In situ hybridization of mouse metaphase chromosomes shows this Hox-6 locus to be located on chromosome 14 (14E2). Hox-6.1 has been studied in detail and the predicted protein sequence of the homeobox is 100% homologous to the Xenopus Xeb1 (formally AC1) homeobox and the human c8 homeobox (Carrasco et al. 1984; Boncinelli et al. 1985; Simeone et al. 1987). Southern blotting shows that the DNA sequence encoding Hox-6.1 is single copy. Expression of Hox-6.1 has been studied in adult tissues and embryos by RNase protection assays, Northern blotting analysis and in situ hybridization. RNase protection assays show that Hox-6.1 transcripts are present in embryos between days 9 1/2 and 13 1/2 of gestation and in extraembryonic tissues at day 9 1/2. Adult expression is detectable in kidney and testis but not in liver, spleen and brain. One major transcript is detectable on Northern blots of kidney and day-13 1/2 embryo RNA. In kidney, this transcript is 2.7 kb whereas in embryos the major transcript is smaller at 1.9 kb, a much fainter band being visible at 2.7 kb. Localized expression of Hox-6.1 is observed in the spinal cord and prevertebral column of day-12 1/2 embryos, and in the posterior mesoderm and ectoderm of day-8 1/4 embryos. An anterior boundary of expression is located just behind the hindbrain whereas the boundary in the mesoderm is located at the level of the 7th prevertebra.
Amino Acid Sequence↗
Effects of isoproterenol on active force and Ca2+ X calmodulin-sensitive phosphodiesterase activity in porcine coronary artery.
Relaxation of vascular smooth muscle following beta-adrenergic stimulation may result from reduction of the cytoplasmic Ca2+ concentration, reduction in the sensitivity of the contractile apparatus to Ca2+, or both. To help resolve these possibilities, we measured the extent of activation of Ca2+ X calmodulin-sensitive phosphodiesterase in intact porcine coronary artery strips as a functional indicator of the cytoplasmic Ca2+ concentration. Both calmodulin-stimulated phosphodiesterase activity and active force increased during K+ stimulation of coronary artery strips. Relaxation of K+-contracted artery strips following stimulus withdrawal was accompanied by rapid inactivation of Ca2+ X calmodulin-sensitive phosphodiesterase. The temporal relationship between isoproterenol-induced relaxation and inactivation of Ca2+ X calmodulin-sensitive phosphodiesterase was studied in both histamine- and K+-contracted tissues. Stimulation of strips with 10 microM histamine or with 44 mM K+ led to comparable increases both in active force and in calmodulin-stimulated phosphodiesterase activity. Thereafter, sustained contraction elicited by histamine was accompanied by a decrease in calmodulin-stimulated phosphodiesterase activity. Isoproterenol rapidly relaxed histamine-contracted strips and accelerated the decrease in phosphodiesterase activity. In contrast, sustained contraction in response to K+ was accompanied by a sustained elevation of calmodulin-stimulated phosphodiesterase activity. Isoproterenol treatment of K+-contracted tissues led to relaxation that was slow and incomplete, and it had very little effect on calmodulin-stimulated phosphodiesterase activity. We conclude that reduction of the cytoplasmic Ca2+ concentration is important for rapid relaxation of the coronary artery following beta-adrenergic stimulation. We cannot disallow the possibility that a decrease in the sensitivity of the contractile apparatus to Ca2+ is involved to some degree.
3',5'-Cyclic-AMP Phosphodiesterases↗
Secretion and processing of staphylococcal nuclease by Bacillus subtilis.
We have studied the secretion and processing of Staphylococcus aureus nuclease in Bacillus subtilis. We show that the initial species of nuclease found in the cell supernatants during short-term radioactive labeling (pulse-chase) had a molecular weight of approximately 18,800 and comigrated in a sodium dodecyl sulfate-polyacrylamide gel with staphylococcal nuclease B. This nuclease B form was processed to the mature nuclease A extracellularly by a phenylmethylsulfonyl fluoride-sensitive protease. The nuclease B-processing site is a consensus signal peptidase site, and the processing of nuclease B was coupled to secretion as judged by pulse-chase experiments. The nuclease A was shown by microsequencing of the N terminus to be 2 amino acid residues shorter than the nuclease A described for S. aureus Foggi. The nuclease B form was still the first species found in the culture supernatant after removal of the N-terminal 26 amino acids of the native 60-amino-acid signal peptide. However, removal of the N-terminal 72 amino acids abolishes secretion of any nuclease form and leads to the intracellular accumulation of nuclease.
Bacillus subtilis↗
Appropriateness of the use of the term sacrifice in experimental animal studies.
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Animals↗
Direct measurements of intramolecular electron transfer rates between cytochrome c and cytochrome c peroxidase: effects of exothermicity and primary sequence on rate.
Rapid mixing of ferrocytochrome c peroxidase [cyt c peroxidase(II)] and ferricytochrome c [cyt c(III)] results in the reduction of cyt c(III) by cyt c peroxidase(II). In 10 mM phosphate, pH 7.0, the rate of decay of cyt c peroxidase(II) and the rate of accumulation of cyt c(II) give equal first-order rate constants: k = 0.23 +/- 0.02 s-1. Equivalent results are obtained by pulse radiolysis using isopropanol radical as the reducing agent. This rate is independent of the initial cyt c(III):cyt c peroxidase(II) ratios. These results are consistent with unimolecular electron transfer occurring within a cyt c(III)-cyt c peroxidase(II) complex. When cyt c is replaced by porphyrin cyt c (iron-free cyt c), a complex still forms with cyt c peroxidase. On radiolysis, using e-aq as the reducing agent, intracomplex electron transfer occurs from the porphyrin cyt c anion radical to cyt c peroxidase(III) with k = 150 s-1. This large rate increase with increasing delta G degrees suggests that the barrier for intracomplex electron transfer is large. Finally, we have briefly investigated how the cyt c peroxidase(II)----cyt c(III) rate depends on the primary structure of cyt c(III). We find the reactivity order to be as follows: yeast (k = 3.4 s-1) greater than horse (k = 0.3 s-1) greater than tuna (k = 0.2 s-1). These results mirror a report [Ho, P. S., Sutoris, C., Liang, N., Margoliash, E. & Hoffman, B. M. (1985) J. Am. Chem. Soc. 107, 1070-1071] on excited state reactions of the cyt c/cyt c peroxidase couple.
Amino Acid Sequence↗
Phorbol diesters alter the contractile responses of porcine coronary artery.
We have studied the effects of activators of the Ca++- and phospholipid-dependent enzyme protein kinase C on isometric tension development by both intact and skinned coronary artery strips. The intact strips contracted upon incubation with 12-O-tetradecanoylphorbol-13-acetate. 12-O-tetradecanoylphorbol-13-acetate produced a leftward shift in the concentration-response relationship for contraction of the tissues by K+, histamine and norepinephrine. Phorbol-12,13-dibutyrate elicited contraction of detergent-skinned artery strips when the free Ca++ concentration in the bathing media was 0.1 microM or greater. This effect was diminished greatly in the presence of polymyxin B, a putative inhibitor of protein kinase C. Phorbol-12,13-dibutyrate shifted the Ca++ concentration-tension response relationship for the skinned tissue to the left. These results are consistent with a role for protein kinase C in regulating the contractile responses of coronary arterial smooth muscle to a variety of stimuli, at least in part by increasing the sensitivity of the contractile apparatus to Ca++.
Animals↗
Genetic analysis of spontaneous and induced palatal slit in C57BL/6 mice.
The results of experiments designed to elucidate the nature of the genetic factors underlying the palatal slit defect in C57BL/6 mice indicate that probably two loci in females and four in males determine the difference in susceptibility to the spontaneous defect between the C57BL/6 and SWV/BcTa strains of mice. Moreover, it appears that these are not the same genes determining the difference between these same strains in palatal slit response to maternal treatment with triamcinolone. Additional evidence is presented to support the view that in the C57BL/6 strain the triamcinolone-induced palatal slit and cleft palate responses are not correlated.
Animal Husbandry↗
Secretion of staphylococcal nuclease by Bacillus subtilis.
The staphylococcal nuclease (nuc) gene from Staphylococcus aureus has been cloned and expressed in Bacillus subtilis. The nuclease protein was expressed either from its own promoter and translation start signals, or from a combination of a B. subtilis promoter, ribosome binding site, and a signal peptide sequence. Greater than 80% of the active gene product was secreted into the medium, whereas, when a signal peptide sequence was absent, as little as 4% of the nuclease activity was found in the culture medium. Intracellular (or cell-bound) nuclease, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting, was shown to have the molecular weight of the predicted precursor protein with the signal peptide. Levels of nuclease reached 50 mg per liter in the culture medium, depending on the growth medium and the strain used. These findings indicate the prospective use of nuclease as a model system for studying secretion of heterologous proteins in B. subtilis.
Bacillus subtilis↗
A new complication of AIDS: thoracic myelitis caused by herpes simplex virus.
Progressive thoracic myelopathy occurred in a patient with AIDS. Concurrent opportunistic infections included disseminated systemic cytomegalovirus, aspergillosis, and cutaneous herpes simplex virus (HSV). At autopsy, immune stains indicated that the myelopathy was caused by HSV type 2 infection of the spinal cord.
Acquired Immunodeficiency Syndrome↗
Living together: grandparents and children tell their problems.
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Adolescent↗
Host specificity of filamentous, segmented microorganisms adherent to the small bowel epithelium in mice and rats.
Germfree rats and mice were given by gavage samples of ileal homogenates prepared from conventional rats and mice. Filamentous, segmented procaryotes adhered to the small bowel epithelium in the ex-germfree mice only when the homogenate was made from mouse bowel and in the ex-germfree rats only when the inoculum came from rats. Thus, the filamentous microorganisms are host animal specific.
Animals↗
Neuropathy and anti-MAG antibodies without detectable serum M-protein.
Anti-MAG IgM antibodies were detected by ELISA in a patient with slowly progressive peripheral neuropathy. Serum IgM content was normal, and no M-protein was detected by serum protein electrophoresis, immunoelectrophoresis, or immunostaining. By immunoblot analysis, the anti-MAG antibodies were IgMk; they reacted with human and bovine MAG but not with mouse MAG. The data suggest that there was an anti-MAG IgM M-protein in concentration too low to be detected by conventional techniques. Tests for anti-MAG antibodies should be done in patients with slowly progressive neuropathy of unknown etiology, even in the absence of detectable serum M-protein.
Aged↗
Neurologic complications in acquired immunodeficiency syndrome (AIDS).
Neurologic syndromes in AIDS are of four types: infections, para-infections, neoplastic, and paraneoplastic. All levels of the neuraxis can be affected. Neurologic complications may be the initial symptom or the cause of death. Aggressive evaluation, including biopsy of cerebral lesions, is indicated because effective treatment can be given.
Acquired Immunodeficiency Syndrome↗
Sensitivity to contact lens solutions.
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Adult↗
Transcription of the 'non-transcribed' spacer of Drosophila melanogaster rDNA.
We have detected a set of transcripts in Drosophila melanogaster cells which are homologous to repeating elements within the 'non-transcribed' spacer region of rDNA. The RNA molecules range from 240 to 1680 nucleotides, differing in length by an integral value of 240 nucleotides. We have sequenced several AluI fragments which characterise the main 240 nucleotide repeating element. We find that each of these fragments contain a segment of approximately 50 nucleotides, which is homologous to the transcription initiation site for pre-rRNA.
Animals↗
Occurrence of oligoclonal bands in multiple sclerosis and other CNS diseases.
The results of cerebrospinal fluid agarose gel electrophoresis in 300 consecutive patients were correlated with neurological examinations and diagnoses, other cerebrospinal fluid studies, and the results of evoked potential examinations. The presence of oligoclonal bands was the most sensitive test for multiple sclerosis (MS); bands were present in from 100% (11/11) of patients with definite MS to 82% (27/33) of those with possible MS (classified by McAlpine criteria). The visual evoked response was the next most sensitive study. Thirty-eight patients without MS or related disorders had bands in the IgG region. Three patients had plasma cell dyscrasias. Seven patients had thick single bands. Single bands did not correlate with chronic polyneuropathy but appeared to be an artifact of storage. Twenty-eight patients had active neurological disease, including cerebral infarction (in 5), viral infection (in 4), remote effect of carcinoma (in 4), and acute and chronic polyneuropathies (in 6). In acute illnesses (i.e., vascular insults), repeat electrophoresis showed disappearance of bands. In continually active disease, bands persisted. These results indicate that the presence of oligoclonal bands provides sensitive supporting evidence for the diagnosis of MS but that bands may be present in other disorders, including those not directly related to infection or abnormal immune response. The data suggest that oligoclonal bands may represent an immune response to neurological injury that is prominent in disorders with a particularly intense or continuous antigenic stimulus.
Central Nervous System Diseases↗