Biomedical subjects
J R Pierce
Publications and source records attributed to J R Pierce.
Recognition and repair of 2-aminofluorene- and 2-(acetylamino)fluorene-DNA adducts by UVRABC nuclease.
Recognition of damage induced by N-hydroxy-2-aminofluorene (N-OH-AF) and N-acetoxy-2-(acetylamino)fluorene (NAAAF) in both phi X174 RFI supercoiled DNA and a linear DNA fragment by purified UVRA, UVRB, and UVRC proteins was investigated. We have previously demonstrated that N-OH-AF and NAAAF treatments produce N-(deoxyguanosin-8-yl)-2-aminofluorene (dG-C8-AF) and N-(deoxyguanosin-8-yl)-2-(acetylamino)fluorene (dG-C8-AAF), respectively, in DNA. Using a piperidine cleavage method and DNA sequence analysis, we have found that all guanine residues can be modified by N-OH-AF and NAAAF. These two kinds of adducts have different impacts on the DNA helix structure; while dG-C8-AF maintains the anti configuration, dG-C8-AAF is in the syn form. phi X174 RF DNA-Escherichia coli transfection results indicate that while the uvrA, uvrB, and uvrC gene products are needed to repair dG-C8-AAF, the uvrC, but not the uvrA or uvrB gene products, is needed for repair of dG-C8-AF. However, we have found that in vitro the UVRA, UVRB, and UVRC proteins must work in concert to nick both dG-C8-AF and dG-C8-AAF. In general, the reactions of UVRABC nuclease toward dG-C8-AF are similar to those toward dG-C8-AAF; it incises seven to eight nucleotides from the 5' side and three to four nucleotides from the 3' side of the DNA adduct. Evidence is presented to suggest that hydrolysis on the 3' and 5' sides of the damaged base by UVRABC nuclease is not simultaneous and that at least occasionally hydrolysis occurs only on the 3' side or on the 5' side of the damage site. The possible mechanisms of UVRABC nuclease incision for AF-DNA are discussed.
Pharmacokinetics of intravenous immunoglobulin in neonates.
Intravenous immunoglobulin (IVIG) may be a therapeutic adjunct to antibiotic treatment of neonatal infections. We examined the pharmacokinetics and safety of IVIG in human neonates. Thirty neonates with suspected sepsis were randomly assigned either to a treatment (receiving either 250, 500, or 1,000 mg/kg of IVIG plus antibiotics) or control (antibiotics alone) group. The 500 mg/kg dose produced a rise in total IgG for greater than 8 and in group B streptococcus (GBS) type-specific IgG for greater than 4-14 days. The type-specific antibody elevation varied with the amount of pathogen-specific antibody and dose of IVIG. Pharmacokinetic analysis suggests a Vdss of 42 ml/kg, Cl of 3.0 ml/kg/day, a biphasic elimination curve, and a terminal elimination half-life of 24.2 days. No toxicity was observed. These data may be valuable in determining optimal dosing schedules for IVIG in treating or preventing neonatal infections.
The Department of Military Medicine--a graduate medical education idea whose time has come.
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Biochemical analysis of spontaneous fepA mutants of Escherichia coli.
The fepA gene of Escherichia coli encodes the outer-membrane receptor protein for ferrienterobactin. Previous genetic studies indicated that fepA mutations occur frequently and suggested that most of the mutations were deletions. In this work seven spontaneous fepA mutations were analyzed by enzyme assay (enterobactin synthase and enterobactin esterase) and by DNA hybridization studies. In two strains, UT500 and UT700, the mutations were confined to the fepA gene. In the remaining mutants, the mutations were large deletions; in several cases, 27 kb or more of DNA had been lost. The deletions, all of which eliminated approximately the left half of the enterobactin gene cluster, extended from the vicinity of the fepC gene counterclockwise into the chromosome. A minimum of three clockwise endpoints were identified and at least two counterclockwise endpoints were detected. The variation in endpoints among the deletions argues against the involvement of a normal transposon in their formation. Also, unexpected homology was found between enterobactin gene cluster DNA and lacPOZ and pSC101.
Escherichia coli K-12 envelope proteins specifically required for ferrienterobactin uptake.
Escherichia coli genes specifically required for transport of iron by the siderophore enterobactin are designated fep. The studies reported here were initiated to identify and localize the fepB product. The plasmid pCP111, which consisted of an 11-kilobase E. coli DNA fragment containing fepB ligated to pACYC184, was constructed. The fepB gene was subcloned; in the process, complementation tests and Tn5 mutagenesis results provided evidence for the existence of a new fep gene, fepC. The order of the transport genes in the ent gene cluster is as follows: fepA fes entF fepC fepB entE. Minicell, maxicell, and in vitro DNA-directed protein synthesizing systems were used to identify the fepB and fepC products. The fepC polypeptide was 30,500 daltons in standard sodium dodecyl sulfate-polyacrylamide gels. The fepB gene was responsible for the appearance of three or four bands (their apparent molecular weights ranged from 31,500 to 36,500) in sodium dodecyl sulfate-polyacrylamide gels, depending on the gel system employed. The largest of these was tentatively designated proFepB, since it apparently had a leader sequence. Localization experiments showed that FepC was a membrane constituent and that mature FepB was present in the periplasm. An additional polypeptide (X) was also encoded by the bacterial DNA of pCP111, but its relationship to iron transport is unknown. The results indicated that ferrienterobactin uptake is mediated by a periplasmic transport system and that genes coding for outer membrane (fepA), periplasmic (fepB), and cytoplasmic membrane (fepC) components have now been identified.
Developmental follow-up of military dependents requiring neonatal intensive care.
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Neonatal intensive care at Fitzsimons Army Medical Center.
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Two fep genes are required for ferrienterochelin uptake in Escherichia coli K-12.
Escherichia coli mutants defective in the assimilation of iron from ferrienterochelin were isolated and characterized. One mutant was able to bind ferrienterochelin to its outer membrane but could not transport it into the cell. Complementation tests with lambda hybrid phage were employed to distinguish the defective gene, which we term fepB, from fepA, the structural gene for the outer membrane ferrienterochelin receptor protein. These same physiological and genetic tests were employed to tentatively classify several previously described fep mutants as carrying either fepA or fepB. The data demonstrate the existence of fepB and provide an explanation for previous difficulties in identifying fepB mutants.
Interference in a chromogenic alpha-amylase assay caused by dye-labeled oligosaccharide-induced precipitation of lipoprotein.
We examined the mechanism by which hypertriglyceridemia interferes with the Roche Diagnostics "Amylochrome" procedure for measurement of amylase in lactescent samples with normal and above-normal amylase activity. The serum blank recommended in the Amylochrome protocol to compensate for lactescence remains inappropriately turbid, compared with the test, and is partly responsible for the underestimation of amylase activity. Other major interference is seen when lipoprotein in hyperlipemic samples with above-normal amylase activity reacts with the soluble oligosaccharide-triazinyl dye product of the Amylochrome assay to form a flocculent blue precipitate. The formation of this precipitate in the test, and its removal, diminish lactescence due to lipemia, an effect that cannot be matched by any manipulation of a serum blank procedure. Oligosaccharide-dye product is removed as a component of the precipitate.
Saturday Conference: stuporous alcoholics: metabolic considerations.
Alcoholics are predisposed to certain metabolic disorders that cause stupor and coma. These entities include acute ethanol intoxication, ethanol-induced hypoglycemia, alcoholic ketoacidosis, ethylene glycol and methanol intoxications, thiamine deficiency, and hepatic encephalopathy. The recognition and management of these entities and the evaluation of stuporous alcoholics are discussed.
Notes on a medical internship.
Though much is currently written about the fate of the medical internship, only one previous study detailing the specifics of the internship experience has been recorded in the literature. The present report summarizes a straight medical internship in a university training program. There were 329 admissions in 123 admitting days. A large variety of medical problems were encountered and a high percentage of patients were acutely ill. Heart disease and cancer accounted for 40% of all admissions and nearly half of all deaths. The most frequently performed procedure was lumbar puncture. It is hoped that this report will stimulate objective record-keeping by others, so that future discussions about the medical internship will be based on fact and not impression.
Extreme eosinophilia and strongyloidiasis: an uncommon manifestation of a common disease.
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Propranolol and retroperitoneal fibrosis.
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A response to: low plus lenses and visual performance: a critical review.
It is contended that the Keller/Amos review is based upon a serious misapplication of statistics or misinterpretation of statistical results, which do not support their conclusions. Proper evaluation supports conclusions diametrically opposed to theirs when based upon the original data of the experiment.
Picture of the month: enteric duplication cyst.
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ST depression suggesting subendocardial ischemia in neonates with respiratory distress syndrome and patent ductus arteriosus.
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Cholesterol embolism: diagnosis antemortem by bone marrow biopsy.
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