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Biomedical subjects

L A Miller

Publications and source records attributed to L A Miller.

At least 127 records · Page 7Linked to original sources

Cloning, expression, and sequence determination of a bacteriophage fragment encoding bacteriophage resistance in Lactococcus lactis.

A number of host-encoded phage resistance mechanisms have been described in lactococci. However, the phage genome has not been exploited as a source of additional resistance determinants. A 4.5-kb BamHI-HindIII fragment of phage nck202.50 (phi 50) was subcloned in streptococcus-Escherichia coli shuttle plasmid pSA3 and introduced into Lactococcus lactis NCK203 and MG1363 by protoplast transformation. This cloned phage fragment directed a bacteriophage resistance phenotype designated Per (phage-encoded resistance). Both phi 50 and a distantly related phage, nck202.48 (phi 48), formed small plaques on strain NCK213 at a slightly reduced efficiency of plaquing on the Per+ host. The per locus was further reduced to a 1.4-kb fragment through in vitro deletion analysis. The 1.4-kb fragment was sequenced, and the Per phenotype was found to be associated with a ca. 500-bp region rich in direct and inverted repeats. We present evidence that the Per region contains a phage origin of replication which, in trans, may interfere with phage replication by titration of DNA polymerase or other essential replication factors. It was demonstrated that the Per+ phenotype is not a result of reduced adsorption or action of a restriction and modification system. Per+ activity was not detected against six independent phages which were previously shown to be sensitive to the Hsp+ mechanism. The mutually exclusive resistance mechanisms could be combined to confer resistance to both types of phages (Hsp resistant and Per resistant) in a single host. This is the first description in lactococci of a phage resistance phenotype, other than superinfection immunity, originating from a lactococcal phage genome.

Amino Acid Sequence↗

5'-Heterogeneity in human progesterone receptor transcripts predicts a new amino-terminal truncated "C"-receptor and unique A-receptor messages.

Human progesterone receptors (PR) are thought to comprise two naturally occurring hormone-binding proteins: 94-kDa A-receptors and 120-kDa B-receptors. In this paper we present evidence for a third human PR, an N-terminally truncated, 45- to 50-kDa species, termed the C-receptor. To determine the translational origin of B- and A-receptors we mapped the multiple messages that code for human PR by Northern blot analyses, using a series of oligonucleotides and cDNA fragment probes corresponding to different regions of the PR message. In addition to the six transcripts of 2.5, 3.2, 4.5, 5.2, 6.1, and 11.4 kilobases (kb) originally described, we found that the 11.4-kb species is a complex of four bands that we have termed I-IV. Analysis of poly(A)+ RNA derived from T47Dv human breast cancer cells using a variety of 5'-specific probes has identified three separate structural classes of human PR transcripts, indicating extensive 5'-termini heterogeneity. Class A messages, the 2.5- and 5.2-kb species, lack the sequences surrounding AUGB (codon 1), which is the translation initiation site for B-receptors, but contain AUGA (codon 165), the initiation site for A-receptors, and, therefore, potentially encode only the latter. Class B messages, consisting of the 3.2-, 4.5-, and 6.1-kb species as well as bands I and II of the 11.4-kb complex contain both AUGB and AUGA and could encode both receptor forms.(ABSTRACT TRUNCATED AT 250 WORDS)

Base Sequence↗

Self-reports of salt intake by 10- to 18-year-olds: relationship to urinary sodium excretion.

Our data indicated that self-reports of consumption of salty foods by children and adolescents were associated with 24-hour urinary sodium excretion. Specifically, youths 10 to 18 years of age who selected a poster depicting high-sodium foods excreted significantly more sodium than youths who selected a poster depicting low-sodium foods. Future research is needed to refine simplified self-report measures, to corroborate the validity of the measures, and to extend the studies to other samples, e.g., younger children.

Adolescent↗

Uric acid excretion in children with urolithiasis.

Urinary uric acid excretion was assessed in 38 children to determine whether hyperuricuria was a risk factor in children with urolithiasis. Uric acid excretion (measured per deciliter glomerular filtration rate), and fractional excretion of uric acid were similar in 27 children with hypercalciuria and calcium oxalate urinary stones, in six children with idiopathic calcium oxalate urolithiasis, and in five with uric acid urolithiasis, of whom four were white boys and one was an Asian girl. One boy with a urate stone had cystinosis. Serum uric acid concentrations exceeded 6.0 mg/dl (360 mumol/L) in two children with hypercalciuria and in two patients with idiopathic calcium oxalate urolithiasis. None of the children with calcium urolithiasis had excessive urinary excretion of uric acid. In children with hypercalciuria, uric acid excretion did not change significantly when dietary sodium was increased from 1.0 to 5.0 gm/1.73 m2. We conclude that excessive urinary uric acid excretion is seldom an additional risk factor in children with calcium urolithiasis and that dietary sodium chloride does not have a strong influence on urinary excretion of uric acid in children with hypercalciuria.

Adolescent↗

Urinary volume in children with urolithiasis.

Urinary volume in 24-hour urine collections was examined in 50 children with hypercalciuria and urolithiasis or hematuria, 12 with idiopathic calcium oxalate urolithiasis and 36 healthy children. Urinary volume was 22.2 +/- 2.0 ml. per kg. per day in healthy children and 25.4 +/- 2.0 ml. per kg. per day in children with hypercalciuria, and it was similar in children with absorptive and renal hypercalciuria, and significantly lower in children with idiopathic calcium oxalate urolithiasis (12.2 +/- 1.4 ml. per kg. per day, p less than 0.001 from controls and children with hypercalciuria). Volume was not statistically different in hypercalciuric children with and without urolithiasis. Urinary sodium excretion in children with idiopathic calculi was not statistically different from controls. Urine osmolality was similar among the groups. Urinary volume represents a risk factor in children with idiopathic calcium oxalate urolithiasis, and increased fluid intake should be emphasized in such patients.

Calcium↗

Renal function in children with idiopathic hypercalciuria.

Abnormalities in renal tubular function have been reported in adult patients with idiopathic renal hypercalciuria. To determine if such abnormalities are present early in the natural history of renal hypercalciuria, we evaluated renal tubular function in ten children with idiopathic renal hypercalciuria, aged 5-17 years. Seven of the children presented with urolithiasis and three with hematuria. Urinary calcium excretion ranged from 4 to 9 mg/kg per day, (5.2 +/- 0.5, mean +/- SEM) with a mean fasting urinary calcium to creatinine ration of 0.31 +/- 0.03. Studies described in this report were performed after 1 week of ingesting a diet containing 1,000 mg calcium, 3,000 mg sodium, and 100 mg purine. Clearance of creatinine ranged from 84 to 159 ml/min per 1.73 m2. Tm phosphate (mg/100 ml GFR) was normal in each child (mean 4.66 +/- 0.06 mg/100 ml GFR). Fractional excretion of uric acid, sodium and beta-2-microglobulin were also normal in each child. Serum bicarbonate concentrations ranged from 21.5 to 27 mEq/l with a mean of 24.4 +/- 0.5 mEq/l and all patients lowered urinary pH to less than 5.5. Hypotonic diuresis demonstrated normal free water clearance with a mean of 12.8 ml/min per 100 ml Cin. Distal sodium delivery and fractional distal sodium reabsorption were normal with a mean of 13.6 +/- 1.2% and 92.7 +/- 0.5%, respectively. Water deprivation studies demonstrated a range of maximum urinary osmolality from 711 to 1,020 mosmol/kg H2O with a mean of 864 +/- 34 mosmol/kg H2O. Seven healthy children, ingesting an identical study diet, concentrated their urine to a mean of 1,059 +/- 31 mosmol/kg h2O.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗

Evaluation of two commercially available test methods to determine the feasibility of testing for respiratory syncytial virus in a community hospital laboratory.

The feasibility of performing respiratory syncytial virus (RSV) testing in a community hospital laboratory was assessed. The Abbott RSV EIA Kit, an enzyme immunoassay (EIA), and the Ortho Respiratory Syncytial Virus Identification Reagent, a direct fluorescent antibody (DFA) technique, were evaluated in terms of technologist time, result time, cost, and agreement of results. There was little difference in the amount of technologist time required for either method. The DFA technique was shown to be much less expensive than the EIA method and also required at least 50% less time to generate a final result. When compared with an indirect fluorescent antibody technique, the EIA technique was more sensitive (87%) than the DFA method (81%). Both methods showed 100% specificity. Based on the findings of this study, the DFA technique was determined to be the more feasible method for a community hospital laboratory to use for the rapid detection of RSV.

Child↗

Increased serum concentrations of 1,25(OH)2 vitamin D in children with fasting hypercalciuria.

Inappropriately elevated concentrations of 1,25(OH)2 vitamin D in serum appear to be responsible for excessive gastrointestinal absorption of dietary calcium in patients with absorptive hypercalciuria. We have examined serum 1,25(OH)2 vitamin D concentrations in another group of children with hypercalciuria in whom urinary calcium excretion was excessive after an overnight fast. Eleven children with idiopathic fasting hypercalciuria (IH) (urinary calcium excretion greater than 4 mg/kg/24 hr and fasting urinary calcium/urinary creatinine ratio greater than 0.21) and seven healthy children were observed while they were eating a diet containing 1 gm calcium per day. Fasting serum 1,25(OH)2 vitamin D concentrations were elevated in children with IH compared with control values (35.3 +/- 3.2 vs 21 +/- 2 pg/ml, P = 0.003), whereas fasting serum parathyroid hormone, 25-OH vitamin D, phosphorus, and ionized calcium concentrations were similar in the two groups. These data suggest that disordered 1,25(OH)2 vitamin D metabolism occurs in children with fasting IH. Absorptive and fasting IH may represent a spectrum of a single disorder characterized by excessive urinary calcium excretion and inappropriately elevated serum concentrations of 1,25(OH)2 vitamin D.

Adolescent↗

Analyzing aversiveness of denatonium saccharide and quinine in rats.

Denatonium saccharide is reported to be the most bitter substance currently known. Two experiments comparing the suppressive capabilities of this compound and the more common bitter, quinine, are presented. Analysis indicated that rats preferred to consume denatonium rather than quinine when afforded a choice between the two. Exp. 2 also indicated that the pairing of quinine with vanilla and almond flavors resulted in subsequent refusal of these flavors. Pairing flavors with denatonium did not produce comparable refusals. Caution is expressed with regard to the use of denatonium saccharide as a rodent repellent.

Animals↗

Increased urinary excretion of renal N-acetyl-beta-glucosaminidase in hypercalciuria.

Urinary excretion of N-acetyl-beta-glucosaminidase (NAG), a lysosomal enzyme, was examined in 33 children with hypercalciuria. Urinary NAG excretion in 13 healthy children was 5.84 +/- 9.35 nmole/hr/mg of creatinine (NAG/Cr) (mean +/- SD) compared with 35.61 +/- 42.04 nmole/hr/mg of creatinine in 23 children with renal hypercalciuria, and 28.99 +/- 13.69 nmole/hr/mg of creatinine in ten children with absorptive hypercalciuria. In children with renal hypercalciuria, NAG/Cr excretion was not statistically different between children with either urolithiasis or hematuria without calculi. In six children with renal hypercalciuria, no significant change in NAG/Cr excretion occurred after a mean duration of 25 weeks of hydrochlorothiazide therapy although urinary calcium to creatinine ratios (UCa/Cr) decreased from 0.24 +/- 0.11 to 0.16 +/- 0.11. We conclude that increased urinary calcium excretion produces renal tubular injury and that the renal injury may not be reversed by short-term alterations in urinary calcium excretion.

Adolescent↗

Hypercalciuria in children with juvenile rheumatoid arthritis: association with hematuria.

After discovering juvenile rheumatoid arthritis (JRA), hematuria, and urolithiasis associated with hypercalciuria in two children, urinary calcium excretion was examined in 38 patients with JRA. Fasting urine calcium/creatinine (mg/mg) (UCa/UCr) ratios were increased (greater than 0.21) in 12 patients, who had a mean UCa/UCr ratio of 0.34 +/- 0.14, compared with 0.09 +/- 0.06 in 26 normocalciuric patients with JRA. Increased UCa/UCr ratios were found more frequently in patients with systemic JRA (P less than 0.05); however, no relationship between UCa/UCr ratios and either functional classification or drug therapy was observed. Four children with increased urine calcium to creatinine ratios were examined more extensively. Twenty-four-hour urine calcium excretion ranged from 4.0 to 7.2 mg/kg/24 hours. An orally administered calcium loading test demonstrated fasting hypercalciuria after dietary calcium restriction in these four patients. Serum calcium, bicarbonate, phosphorus, and parathyroid hormone values were normal. Hematuria was found in six of 12 hypercalciuric patients with JRA but in only three of 26 normocalciuric patients (P less than 0.016). We conclude that urinary calcium excretion is frequently increased in patients with JRA and that hypercalciuria may be related to the pathogenesis of hematuria in some of them.

Arthritis, Juvenile↗