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R A Preston

Publications and source records attributed to R A Preston.

At least 19 recordsLinked to original sources
PubMed1992

The VPH1 gene encodes a 95-kDa integral membrane polypeptide required for in vivo assembly and activity of the yeast vacuolar H(+)-ATPase.

Yeast vacuolar acidification-defective (vph) mutants were identified using the pH-sensitive fluorescence of 6-carboxyfluorescein diacetate (Preston, R. A., Murphy, R. F., and Jones, E. W. (1989) Proc. Natl. Acad. Sci. U.S.A. 86, 7027-7031). Vacuoles purified from yeast bearing the vph1-1 mutation had no detectable bafilomycin-sensitive ATPase activity or ATP-dependent proton pumping. The peripherally bound nucleotide-binding subunits of the vacuolar H(+)-ATPase (60 and 69 kDa) were no longer associated with vacuolar membranes yet were present in wild type levels in yeast whole cell extracts. The VPH1 gene was cloned by complementation of the vph1-1 mutation and independently cloned by screening a lambda gt11 expression library with antibodies directed against a 95-kDa vacuolar integral membrane protein. Deletion disruption of the VPH1 gene revealed that the VPH1 gene is not essential for viability but is required for vacuolar H(+)-ATPase assembly and vacuolar acidification. VPH1 encodes a predicted polypeptide of 840 amino acid residues (molecular mass 95.6 kDa) and contains six putative membrane-spanning regions. Cell fractionation and immunodetection demonstrate that Vph1p is a vacuolar integral membrane protein that co-purifies with vacuolar H(+)-ATPase activity. Multiple sequence alignments show extensive homology over the entire lengths of the following four polypeptides: Vph1p, the 116-kDa polypeptide of the rat clathrin-coated vesicles/synaptic vesicle proton pump, the predicted polypeptide encoded by the yeast gene STV1 (Similar To VPH1, identified as an open reading frame next to the BUB2 gene), and the TJ6 mouse immune suppressor factor.

Amino Acid Sequence↗
PubMed1992

Antihypertensive drug therapy does not perturb the circadian blood pressure pattern.

The literature was reviewed to determine whether failure of older antihypertensive drugs to suppress the early morning increase of blood pressure was responsible, in part, for the less than satisfactory rate of reduction in cardiovascular mortality rates in the United States. The authors found that neither the old nor the new antihypertensive drugs altered the 24-hour blood pressure curve pattern, although long-acting drugs did show continued effect at the end of the 24-hour period when compared with placebo. The efficacy of these drugs most likely lies with their blood pressure lowering and other ancillary effects and not with pattern changes. More importantly, examination of new data (1989) shows that the rate of decline in death due to diseases of the heart has exceeded that for cerebrovascular diseases. Our overall health care effort may be more successful than we thought.

Antihypertensive Agents↗
PubMed1992

Genes required for vacuolar acidity in Saccharomyces cerevisiae.

Mutations that cause loss of acidity in the vacuole (lysosome) of Saccharomyces cerevisiae were identified by screening colonies labeled with the fluorescent, pH-sensitive, vacuolar labeling agent, 6-carboxyfluorescein. Thirty nine vacuolar pH (Vph-) mutants were identified. Four of these contained mutant alleles of the previously described PEP3, PEP5, PEP6 and PEP7 genes. The remaining mutants defined eight complementation groups of vph mutations. No alleles of the VAT2 or TFP1 genes (known to encode subunits of the vacuolar H(+)-ATPase) were identified in the Vph- screen. Strains bearing mutations in any of six of the VPH genes failed to grow on medium buffered at neutral pH; otherwise, none of the vph mutations caused notable growth inhibition on standard yeast media. Expression of the vacuolar protease, carboxypeptidase Y, was defective in strains bearing vph4 mutations but was apparently normal in strains bearing any of the other vph mutations. Defects in vacuolar morphology at the light microscope level were evident in all Vph- mutants. Strains that contained representative mutant alleles of the 17 previously described PEP genes were assayed for vacuolar pH; mutations in seven of the PEP genes (including PEP3, PEP5, PEP6 and PEP7) caused loss of vacuolar acidity.

Alleles↗
PubMed1991

Isolation and characterization of PEP3, a gene required for vacuolar biogenesis in Saccharomyces cerevisiae.

The Saccharomyces cerevisiae PEP3 gene was cloned from a wild-type genomic library by complementation of the carboxypeptidase Y deficiency in a pep3-12 strain. Subclone complementation results localized the PEP3 gene to a 3.8-kb DNA fragment. The DNA sequence of the fragment was determined; a 2,754-bp open reading frame predicts that the PEP3 gene product is a hydrophilic, 107-kDa protein that has no significant similarity to any known protein. The PEP3 predicted protein has a zinc finger (CX2CX13CX2C) near its C terminus that has spacing and slight sequence similarity to the adenovirus E1a zinc finger. A radiolabeled PEP3 DNA probe hybridized to an RNA transcript of 3.1 kb in extracts of log-phase and diauxic lag-phase cells. Cells bearing pep3 deletion/disruption alleles were viable, had decreased levels of protease A, protease B, and carboxypeptidase Y antigens, had decreased repressible alkaline phosphatase activity, and contained very few normal vacuolelike organelles by fluorescence microscopy and electron microscopy but had an abundance of extremely small vesicles that stained with carboxyfluorescein diacetate, were severely inhibited for growth at 37 degrees C, and were incapable of sporulating (as homozygotes). Fractionation of cells expressing a bifunctional PEP3::SUC2 fusion protein indicated that the PEP3 gene product is present at low abundance in both log-phase and stationary cells and is a vacuolar peripheral membrane protein. Sequence identity established that PEP3 and VPS18 (J. S. Robinson, T. R. Graham, and S. D. Emr, Mol. Cell. Biol. 11:5813-5824, 1991) are the same gene.

Adaptor Proteins, Vesicular Transport↗
PubMed1991

Conditioned reinforcement value and choice.

The delay-reduction hypothesis of conditioned reinforcement states that the reinforcing value of a food-associated stimulus is determined by the delay to primary reinforcement signaled by the onset of the stimulus relative to the average delay to primary reinforcement in the conditioning situation. In contrast, most contemporary models of conditioned reinforcement strength posit that the reinforcing strength of a stimulus is some simple function only of the delay to primary reinforcement in the presence of stimulus. The delay-reduction hypothesis diverges from other conditioned reinforcement models in that it predicts that a fixed-duration food-paired stimulus will have different reinforcing values depending on the frequency of its presentation. In Experiment 1, pigeons' key pecks were reinforced according to concurrent-chains schedules with variable-interval 10-second and variable-interval 20-second terminal-link schedules. The initial-link schedule preceding the shorter terminal link was always variable-interval 60 seconds, and the initial-link schedule requirement preceding the longer terminal link was varied between 1 second and 60 seconds across conditions. In Experiment 2, the initial-link schedule preceding the longer of two terminal links was varied for each of three groups of pigeons. The terminal links of the concurrent chains for the three groups were variable-interval 10 seconds and 20 seconds, variable-interval 10 seconds and 30 seconds, and variable-interval 30 seconds and 50 seconds. In both experiments, preference for the shorter terminal link was either a bitonic function or an inverse function of the initial-link schedule preceding the longer terminal-link schedule. Consistent with the predictions of the delay-reduction hypothesis, the relative values of the terminal-link stimuli changed as a function of the overall frequency of primary reinforcement. Vaughan's (1985) melioration model, which was shown to be formally similar to Squires and Fantino's (1971) delay-reduction model, can be modified so as to predict these results without changing its underlying assumptions.

Animals↗
PubMed1991

Choice and conditioned reinforcement.

A potential weakness of one formulation of delay-reduction theory is its failure to include a term for rate of conditioned reinforcement, that is, the rate at which the terminal-link stimuli occur in concurrent-chains schedules. The present studies assessed whether or not rate of conditioned reinforcement has an independent effect upon choice. Pigeons responded on either modified concurrent-chains schedules or on comparable concurrent-tandem schedules. The initial link was shortened on only one of two concurrent-chains schedules and on only one of two corresponding concurrent-tandem schedules. This manipulation increased rate of conditioned reinforcement sharply in the chain but not in the tandem schedule. According to a formulation of delay-reduction theory, when the outcomes chosen (the terminal links) are equal, as in Experiment 1, choice should depend only on rate of primary reinforcement; thus, choice should be equivalent for the tandem and chain schedules despite a large difference in rate of conditioned reinforcement. When the outcomes chosen are unequal, however, as in Experiment 2, choice should depend upon both rate of primary reinforcement and relative signaled delay reduction; thus, larger preferences should occur in the chain than in the tandem schedules. These predictions were confirmed, suggesting that increasing the rate of conditioned reinforcement on concurrent-chains schedules may have no independent effect on choice.

Animals↗
PubMed1991

Classic therapeutic trials in hypertension. Were patients vulnerable to unsuppressed peak morning blood pressure?

A metaanalysis of the major clinical trials on the benefits of antihypertensive therapy revealed that there is an overall reduction of coronary events by treatment, but that it is only about one-half of the reduction predicted. The reduction in stroke achieved the predicted number. This "failure" has been attributed to study design, patient age at entry, preexisting disease, high-lipid diets, drug-associated cardiac dysrhythmias, and the "J-curve" effect. It is also possible that the drugs used in these trials did not suppress the morning acceleration of blood pressure during which most coronary and cerebrovascular events occur. Ambulatory blood pressure monitoring studies suggest that antihypertensive drugs reduce overall blood pressure throughout the 24-h period, but do not change the circadian curve. The beneficial effect of antihypertensive therapy probably rests with the magnitude of blood pressure reduction. It is unlikely that lack of suppression of morning acceleration alone accounted for the failure of "older" drugs to prevent as much cardiac mortality as expected.

Aged↗
PubMed1990

Facial nerve injury and hemifacial spasm.

We studied evidence of facial nerve damage in patients with hemifacial spasm. Three types of evidence of nerve damage were analyzed: objectively measured weakness in eyelid protractor strength, clinically evident weakness of muscles innervated by the seventh nerve, and clinically evident aberrant seventh nerve regeneration. Of the 60 patients in the study, 54 (90%) had at least one of these features of seventh nerve damage. Objectively measured eyelid protractor weakness was noted in 27 of 58 patients (47%) who were tested. Clinically apparent weakness of at least one of four facial muscle groups was noted in 42 of 60 patients (70%). Aberrant seventh nerve regeneration was documented in 25 of 60 patients (42%). These findings indicate that facial nerve damage is common in patients with hemifacial spasm.

Adult↗
PubMed1990

Renal biopsy in patients 65 years of age or older. An analysis of the results of 334 biopsies.

We studied the clinical and pathological data for 334 patients age 65 or more who underwent renal biopsy for acute renal failure (ARF, n = 55), subacute renal failure (SRF, n = 72), chronic renal failure (CRF, n = 57), proteinuria (n = 137), and hematuria (n = 13). Tissue diagnoses were glomerulopathy (n = 252, 75.4%), acute tubular lesions (n = 18), interstitial nephritis (n = 23), vascular diseases (n = 36, including 14 with cholesterol emboli), and five miscellaneous diagnoses. Of the 55 patients with ARF, 23 had a glomerular lesion, 15 had acute tubular necrosis, and 8 had acute interstitial nephritis. Of 72 patients with SRF, 49 had a glomerulopathy, 12 had a vascular disorder, and six had acute interstitial nephritis. Hence, patients with ARF or SRF exhibited a high potential for reversible lesions. Only 11.3% of patients with CRF had potentially reversible causes. The most common causes of proteinuria were membranous glomerulopathy (34.3%), minimal change disease (14.6%), focal segmental sclerosis (11.7%), and amyloidosis (8.8%). Of the 25 patients with advanced nephrosclerosis, 24 had renal failure, 20 were hypertensive, and 13 had cholesterol emboli. Of 33 patients with diabetes mellitus, 66.7% were found to have lesions not related to diabetes. We conclude that renal biopsy is most useful in older patients with ARF or SRF because of potentially reversible renal disease. Old age alone is not a contraindication to performing a renal biopsy.

Acute Kidney Injury↗
PubMed1990

Role of vacuolar acidification in protein sorting and zymogen activation: a genetic analysis of the yeast vacuolar proton-translocating ATPase.

Vacuolar acidification has been proposed to play a key role in a number of cellular processes, including protein sorting, zymogen activation, and maintenance of intracellular pH. We investigated the significance of vacuolar acidification by cloning and mutagenizing the gene for the yeast vacuolar proton-translocating ATPase 60-kilodalton subunit (VAT2). Cells carrying a vat2 null allele were viable; however, these cells were severely defective for growth in medium buffered at neutral pH. Vacuoles isolated from cells bearing the vat2 null allele were completely devoid of vacuolar ATPase activity. The pH of the vacuolar lumen of cells bearing the vat2 mutation was 7.1, compared with the wild-type pH of 6.1, as determined by a flow cytometric pH assay. These results indicate that the vacuolar proton-translocating ATPase complex is essential for vacuolar acidification and that the low-pH state of the vacuole is crucial for normal growth. The vacuolar acidification-defective vat2 mutant exhibited normal zymogen activation but displayed a minor defect in vacuolar protein sorting.

Amino Acid Sequence↗
PubMed1989

Assay of vacuolar pH in yeast and identification of acidification-defective mutants.

As part of a genetic analysis of the biogenesis and function of the vacuole (lysosome) in the yeast Saccharomyces cerevisiae, assays of vacuolar pH were developed and used to identify mutants defective in vacuolar acidification. Vacuoles were labeled with 6-carboxyfluorescein with the membrane-permeant precursor 6-carboxyfluorescein diacetate. Dual-excitation flow cytometry was used to calibrate the pH-dependence of 6-carboxyfluorescein fluorescence in vivo. Vacuoles in wild-type yeast were mildly acidic, pH 6.2, in cells grown under several different conditions. Cultures labeled with 6-carboxyfluorescein were screened by fluorescence-ratio microscopy to detect mutants that had defects related to vacuolar acidification. A recessive nuclear mutation, vph1-1, caused an abnormally high vacuolar pH of 6.9, as assayed by flow cytometry, and eliminated vacuolar uptake of the weak base quinacrine. Acidification in a pep12::LEU2 mutant appeared defective by fluorescence-ratio microscopy and quinacrine-uptake assays, but the vacuolar pH in the pep12::LEU2 mutant was nearly normal (pH 6.3) in flow cytometric assays.

Hydrogen-Ion Concentration↗
PubMed1989

Newer principles of patient profiling for antihypertensive therapy.

The mosaic of essential hypertension seems to be gaining new tiles at an accelerating rate. Of the many pathophysiological factors and markers for the factors that can be considered as possible elements of a profiling algorithm, there are relatively few with solid data to support their use. Use of plasma renin activity for prospective profiling does not appear to be valid for individual patients. It remains useful for diagnosis of renal artery stenosis and primary aldosteronism. The black race does have some well-documented pathophysiological differences from the white race regarding hypertension, and there are some useful data bits for selecting specific therapy. The elderly, obese patients, and young patients with hyperdynamic circulation appear to have enough group characteristics to enable selection of drugs more targeted to their needs. Anxious patients who often display symptoms that mimic pheochromocytoma symptoms comprise a unique group. One of the largest groups is patients with hypertension plus one or more concomitant diseases. Finally, drug selection decisions must also consider the effect of drugs on serum lipoproteins, left ventricular hypertrophy and vascular compliance, sexual function and other quality of life issues, and cost.

Age Factors↗
PubMed1988

Acute rhabdomyolysis associated with cocaine intoxication.

Because an increasing number of patients were arriving at our emergency room with cocaine intoxication and rhabdomyolysis, we reviewed our experience with such patients. We identified 39 patients seen at our institution over an eight-year period with acute rhabdomyolysis after cocaine use. The patients' mean creatine kinase level was 12,187 U per liter (range, 1756 to 85,000). Thirteen of the 39 patients (33 percent) had acute renal failure; 6 of them died. In comparison to the patients with normal renal function, those with renal failure were more often admitted with profound hypotension (46 vs. 4 percent; P less than 0.001), hyperpyrexia (69 vs. 15 percent; P less than 0.001), and markedly elevated serum creatine kinase levels (mean, 28,084 vs. 7931 U per liter; P less than 0.01). Disseminated intravascular coagulation developed in seven patients with renal failure. All six deaths were in this group. Severe hepatic dysfunction was found in 11 patients with renal failure. We conclude that cocaine intoxication can cause acute rhabdomyolysis with acute renal failure, severe liver dysfunction, and disseminated intravascular coagulation and that the mortality rate among patients with this syndrome is high.

Acute Disease↗
PubMed1988

Functional relationships between L- and D-alanine, inosine and NH4Cl during germination of spores of Bacillus cereus T.

The results of a physiological study of the interaction between NH4Cl, inosine, and the stereoisomers of alanine during germination of spores of Bacillus cereus T are presented. Detailed kinetics for the germination of unheated spores in moderate concentrations of L-alanine (in the absence of auto-inhibition due to alanine racemase) are established, as is the specificity of the stimulatory effect of NH4Cl in relation to other salts, amines, and germinants. The results suggest that NH4Cl and inosine affect an early step in germination closely related to the function of an L-alanine receptor.

Alanine↗
PubMed1988

Choice and foraging: the effects of accessibility on acceptability.

Pigeons responded in a successive-encounters choice procedure in which accessibility of the less profitable of two outcomes varied either in terms of probability of encounter or search time to encounter (keeping search time to the more profitable outcome constant). When the less profitable outcome was made more probable its acceptance became more likely. However, when search time to encounter the less profitable outcome was shortened, its acceptance became less likely. Both results are consistent with the delay-reduction hypothesis and with an optimality model developed for application to the successive-encounters choice procedure.

Animals↗