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Biomedical subjects

J Harder

Publications and source records attributed to J Harder.

54 records · Page 3Linked to original sources

Activation of the anaerobic ribonucleotide reductase from Escherichia coli by S-adenosylmethionine.

The anaerobic ribonucleoside triphosphate reductase from Escherichia coli reduces CTP to dCTP in the presence of a second protein, named dA1, and a Chelex-treated boiled extract of the bacteria, named RT. The reaction requires S-adenosylmethionine, NADPH, dithiothreitol, ATP, and Mg2+ and K+ ions. It occurs only under anaerobic conditions. We now show that the overall reaction occurs in two steps. The first is an activation of the reductase by dA1 and RT and requires S-adenosylmethionine, NADPH, dithiothreitol, and possibly K+ ions. In the second step, the activated reductase reduces CTP to dCTP with ATP acting as an allosteric effector. During activation, S-adenosylmethionine is cleaved reductively to methionine + 5'-deoxyadenosine. This step is inhibited strongly by S-adenosylhomocysteine and various chelators. The activation of the anaerobic reductase shows a considerable similarity to that of pyruvate formate-lyase (Knappe, J., Neugebauer, F. A., Blaschkowski, H. P., and Gänzler, M. (1984) Proc. Natl. Acad. Sci. U.S.A. 81, 1332-1335).

Anaerobiosis↗

Identification of a free radical and oxygen dependence of ribonucleotide reductase in yeast.

Ribonucleotide reductase is a key enzyme for DNA biosynthesis. The enzymes isolated from animal and plant cells possess a stable tyrosyl free radical which is essential for catalysis. Fungal ribonucleotide reductases are little known; the partially characterized enzyme from yeast cells proved exceptionally shortlived, and a free radical could not as yet be demonstrated. We here show that a doublet ESR signal centered at g = 2.0046 can be measured below 60 degrees K in rapidly purified protein samples which is very similar to the ESR spectra of the tyrosine radicals present in other eukaryotic ribonucleotide reductases in structure, microwave saturation, and quenching by hydroxyurea. Because generation of these radicals requires oxygen, anaerobic yeast cultures were also studied. No change in ribonucleotide reductase was observed at 50 ppm residual oxygen in the gas phase, but cell proliferation ceased entirely under complete anaerobiosis.

Blotting, Western↗

Duplex ultrasonography of the fetal aorta, umbilical artery, and placental arcuate artery throughout normal human pregnancy.

A cross-sectional duplex study of blood flow in the fetal aorta, umbilical artery, and arcuate arteries of 111 normal subjects was performed once at between 20 and 42 weeks gestation. Peak systolic velocity, end-diastolic velocity, mean velocity, and volume flow were measured in the fetal aorta, and systolic to diastolic (S/D) ratios were measured in all three vessels. Velocities show an inverse exponential (Y = Exp [1/X x B + C]) increase with gestational age, reaching a plateau just before term, while S/D ratios in the aorta and umbilical arteries show a linear decrease. Such findings are at variance with some published data. Our results suggest a reduction in fetal placental resistance at term and provide useful data concerning normal fetal and placental circulations.

Aorta↗

Pulmonary hypoplasia testing in clinical obstetrics.

An ultrasound-based diagnostic test for predicting pulmonary hypoplasia antenatally by measurements of fetal chest circumference was applied prospectively to 45 patients at risk for developing this disorder. The outcome for the newborn infant and the autopsy findings were correlated with the antenatal prediction. The results of this test demonstrated a sensitivity and a specificity of 88% and 96%, respectively. The application of this test to clinical practice is recommended.

Amniotic Fluid↗

Evaluation of fetal cardiac dysrhythmias with two-dimensional, M-mode, and pulsed Doppler ultrasonography.

Forty-three patients with fetal cardiac dysrhythmia were referred to a tertiary ultrasound department for further evaluation. The patients were subdivided according to the type of dysrhythmia diagnosed. All patients with irregular fetal heart rate had a good perinatal outcome. Of the six patients with fetal tachycardia, two required in utero therapy with maternal digoxin, and one of these fetuses had a cardiac tumor. The eleven patients with fetal bradycardia had the worst perinatal outcome, with four deaths occurring. The results suggest that all patients with fetal tachycardia or bradycardia should have a complete fetal cardiac assessment in a center experienced with the management of such cases.

Algorithms↗

Atrial natriuretic peptide production in association with nonimmune fetal hydrops.

The presence and elevation of atrial natriuretic peptide in fetuses has not previously been demonstrated. This study of right atrial pacing in fetal lambs demonstrated a threefold to fourfold increase in atrial natriuretic peptide during the production of fetal hydrops. Its rate of return to a normal level paralleled the clearance of fetal hydrops. Its possible role in fetal cardiovascular hemodynamics is discussed.

Animals↗

Doppler ultrasound prediction of fetal outcome in twin pregnancies.

The usefulness of pulsed Doppler ultrasound in predicting twin pregnancies destined for unsatisfactory outcomes was examined in 30 twin pregnancies. Fetal flow velocity waveform analysis (systolic/diastolic ratio and pulsatility index) and quantitative analysis in the descending aorta and umbilical artery demonstrated good sensitivity and specificity when compared with biparietal diameter and abdominal circumference measurements. This technique shows some promise as a useful adjunct in the management of twin pregnancies.

Aorta, Thoracic↗

Ultrasound evaluation of tachycardia-induced hydrops in the fetal lamb.

Nonimmune hydrops was induced in six fetal lambs at 120 days' gestation by right atrial pacing at 300 beats/min. Biochemical, hemodynamic, and Doppler parameters were observed and monitored by real-time ultrasound during the creation (18 to 42 hours) and resolution of hydrops. The significant changes seen were hypoproteinemia (34.3 to 29.0 mg/100 ml), venous hypertension (24.6 to 31.3 mm Hg), decreases in thoracic aorta flow, and decreases in systolic/diastolic ratios in the aorta and umbilical artery. These changes were reversed with the discontinuation of pacing and the resolution of fetal hydrops.

Animals↗

Ultrasound prediction of pulmonary hypoplasia.

Normograms for ultrasound-measured fetal chest and heart circumferences were constructed from 83 normal pregnancies of 24 to 39 weeks' duration. Of six cases at risk for pulmonary hypoplasia, all of whom ultimately proved to have the condition, four fell below the fifth percentile for chest circumferences. Both cases in which the chest circumference was within the normal range had pleural effusions.

Cross-Sectional Studies↗

In utero evaluation of fetal cardiac structure: a preliminary report.

Fetal echocardiography as a tool for the prenatal diagnosis of cardiac structural abnormalities was utilized in 27 consecutive patients who were referred for evaluation. Four abnormalities were recognized prenatally and confirmed postnatally. A fifth was identified postnatally. The sensitivity and specificity of this diagnostic test were 80% and 100%, respectively. The utilization of this tool in a tertiary care setting by an adequately trained team has proved to be helpful and is recommended.

Echocardiography↗

Deoxyribonucleotide synthesis in an Escherichia coli mutant (H 1491) which lacks ribonucleotide reductase subunit B2.

An iron-sensitive mutant of E. coli with a Mudl phage insertion in the nrdB gene lacks subunit B2 of the key enzyme of DNA synthesis, ribonucleotide reductase. Nevertheless, these cells are capable of growing in minimal media under anaerobic conditions, indicating a second enzyme or pathway for deoxyribonucleotide synthesis. We here show that ribonucleotide reduction cannot be unambiguously measured in bacterial extracts whereas phosphorylase-catalyzed deoxyribosyl transfer does occur; however these salvage reactions could not function in vivo in the absence of deoxyribosides. It is suggested that the cells possess a specific, anaerobic ribonucleotide reductase which escapes detection under aerobic standard conditions, similar to the situation found in strictly anaerobic methanogens.

Anaerobiosis↗

A student's experience.

The following is the journal kept by Jan Harder, a student in the collaborative nursing program at North Island College, of her work experience in the emergency unit at St. Joseph's Hospital in Comox. It reflects the enthusiasm that students bring to and take away from their Nurses' Work Experience.

Adaptation, Psychological↗