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Biomedical subjects

S Schaefer

Publications and source records attributed to S Schaefer.

At least 73 records · Page 4Linked to original sources
PubMed1993

The conserved nonanucleotide motif of the geminivirus stem-loop sequence promotes replicational release of virus molecules from redundant copies.

Recombinant plasmids containing head-to-tail copies of different coat-protein replacement genomes of wheat dwarf virus (WDV) were used to study the mechanism leading to the release of replicating unit-length molecules in suspension culture cells of Triticum monococcum. For plasmids bearing two complete genomes, the viral unit bracketed by the two large intergenic regions (LIR) becomes preferentially released. Addition of a third copy of the LIR on the inoculum plasmid is necessary for release of both WDV genomes with the same efficiency. Using plasmids containing a single viral genome flanked by two different hybrid LIRs, we show that the sequence TAATATTA, which is part of the conserved geminivirus nonanucleotide motif of the potential hairpin structure, is the region of the LIR within which the release of unit-length molecules occurs. Moreover, the data suggest that this release results primarily from rolling-circle replication, also in situations where intramolecular homologous recombination is simultaneously possible.

Base Sequence↗
PubMed1993

Myocardial adaptation during acute hibernation: mechanisms of phosphocreatine recovery.

OBJECTIVES: Acute hibernation, defined as a prolonged period of moderately reduced oxygen supply and stable haemodynamic function, results in metabolic adaptation characterised primarily by an increase in phosphocreatine. The mechanism of this increase in phosphocreatine is unknown, but has been postulated to result from either an increase in adenosine triphosphate (ATP) production or a decrease in ATP utilisation under conditions of constant myocardial oxygen consumption (MVO2). These experiments were performed to test the hypotheses that (1) acute hibernation could be modelled in an isolated perfused rat heart exhibiting metabolic adaptation; and (2) recovery of phosphocreatine could be explained by alterations in relative creatine kinase flux during hibernation. METHODS: Nuclear magnetic resonance techniques were used in an isolated, perfused rat heart model of acute hibernation to determine the changes in metabolites and creatine kinase kinetics. A flow reduction from 12.5 to 5.4 ml.min-1 was employed for two hours, followed by reperfusion. RESULTS: Reduction of flow resulted in a stable 44% reduction in rate-pressure product. Phosphocreatine had a significant decrease of 9% within the first 15 minutes of ischaemia, but recovered to control values by the end of ischaemia. ATP and [ADP], although unchanged in the early phase of ischaemia, were progressively reduced during the later phase of ischaemia. Intracellular pH fell from 6.99(0.04) to 6.92(0.03) after 15 minutes of ischaemia with little recovery. Saturation transfer measurements showed stability of the forward flux in the creatine kinase reaction during ischaemia, but a progressive reduction in the calculated reverse flux. CONCLUSIONS: These data show that acute hibernation can be modelled in an isolated perfused heart, exhibiting recovery of phosphocreatine despite progressive reductions in ATP. Metabolic changes during acute hibernation have a phasic response characterised by an early ischaemic phase and a later adaptive phase. There is a time related change in measured creatine kinase flux, consistent with a differential change in either ATP production via an increase in MB creatine kinase isoenzyme or a shift in the activity of mitochondrial v cytosolic creatine kinase, a reduction in ATP utilisation via increased efficiency of ATP utilisation at the myofibril, or a changing contribution of glycolytically produced ATP.

Adenosine Diphosphate↗
PubMed1993

Identification of a new hepatitis B virus (HBV) genotype from Brazil that expresses HBV surface antigen subtype adw4.

The complete genome of a hepatitis B virus (HBV) from Brazil that expressed the subtype adw4 of HBV surface antigen (HBsAg) was cloned and sequenced. The genome, termed w4B, consists of 3215 bp. The overall genetic organization of typical hepadnaviruses with four open reading frames including the preC region was found to be conserved. When comparing the w4B sequence with 19 complete HBV genomes it was, however, found to be more divergent (15%) than any other HBV sequence thus far reported. Until now, no more than 11% divergence has been reported. Distinct from the five known HBV genotypes A to E, w4B made up a new, sixth genotype. The importance of the conserved third start codon in the HBV X gene became apparent in isolate w4B. By mutation, this ATG was out of frame, and by what appears to have been a linked mutation, a new start site two codons downstream was re-established. The significance of several other mutations is discussed.

Amino Acid Sequence↗
PubMed1993

Genotyping by multiplex polymerase chain reaction for detection of endemic hepatitis B virus transmission.

A nested polymerase chain reaction (PCR) protocol was developed for rapid genotyping of hepatitis B virus (HBV). During the first PCR round, a universal HBV primer pair was used to amplify the entire pre-S region of the HBV genome. Within the pre-S region, many nucleotide exchanges are observed. These are partly correlated to the serological hepatitis B surface antigen subtypes. Five additional subtype-specific primers were selected from that region which, together with two universal non-group-specific primers, generated specific combinations of two to four DNA fragments of defined sizes. By this approach, 55 hepatitis B surface antigen-positive patients from a pediatric oncology unit in Germany were analyzed. Fifty-four patients who had been infected within 2 years had an identical pattern in the multiplex PCR, suggesting a common source of infection and person-to-person transmission within the unit. One child who was infected 5 years later had a different PCR pattern and, therefore, must have been infected from a different source. Furthermore, 109 serum samples taken from pregnant Cameroonian women and 25 serum samples from their babies taken 6 months after birth were analyzed. In one case, mother-to-infant transmission of the virus was demonstrated. Apart from its role in epidemiological studies on HBV, multiplex PCR may also be a useful tool for rapid genetic analysis in other fields if there is a moderate degree of sequence variation which enables the design of specific primers.

Base Sequence↗
PubMed1993

Comparison of two methods of tonsillectomy.

No consensus exists regarding the best method of tonsillectomy. This report concerns two popular methods: 1. electrocautery excision and 2. dissection/snare followed by point coagulation of bleeding sites. To compare these methods, a prospective, randomized, single-blinded study was conducted in which 28 patients had one tonsil removed by dissection/snare and selective cautery of bleeders and the other removed by the electrocautery. Operative time and blood loss were compared. Patients rated the severity of their pain and blood loss postoperatively. Intraoperative bleeding was significantly less on the side of cautery excision, although the operative time was longer. At follow-up, pain was rated worse on the side of cautery excision.

Adolescent↗
PubMed1992

Metabolic response of the human heart to inotropic stimulation: in vivo phosphorus-31 studies of normal and cardiomyopathic myocardium.

In order to determine if an increase in myocardial oxygen consumption is accompanied by changes in high energy phosphates in normal subjects and patients with dilated cardiomyopathy, phosphorus-31 spectra were acquired under resting conditions and during dobutamine infusion. In seven normal subjects, dobutamine raised the rate-pressure product to 226% of control. The ratio of PCr/ATP was 1.86 +/- 0.17 (mean +/- SE) under resting conditions and 1.90 +/- 0.22 (P = 0.44) with dobutamine infusion. In eight patients with dilated cardiomyopathy, dobutamine raised the rate-pressure product to 161% of control. As in the normal subjects, the ratio of PCr/ATP under resting conditions (1.63 +/- 0.24) was unchanged during dobutamine infusion (1.57 +/- 0.24, P = 0.38). These data indicate that increases in cardiac work do not have a major effect on high energy phosphate concentrations in normal subjects or in patients with clinically compensated dilated cardiomyopathy.

Adenosine Triphosphate↗
PubMed1992

Lifetime diagnoses in patients with somatoform disorders: which came first?

Thirty inpatients with somatoform disorders were examined with the structured clinical interview SCID for psychiatric lifetime diagnosis. In the present diagnoses, we found a concordance of 63% for somatoform and affective disorders and the lifetime comorbidity of both disorders was 87%. Additionally, patients with somatoform disorders frequently had a history of other psychiatric disorders (for example, anxiety disorders, 40%). For 73% of patients with somatoform disorders and a history of affective disorders, the onset of the somatoform disorder was prior to the onset of another psychiatric disorder. The time interval between the onsets of somatoform disorders and affective disorders was greater than 1 year for most patients; for 46% of the patients with a history of both disorders, the time interval between the two disorders was more than 5 years. The course of illness for somatoform and affective disorders was quite different; while affective disorders tended to episodic periods with interim remissions, the somatoform disorders usually showed long, chronic courses (mean duration of the current somatoform disorder was 11.9 years). Finally, the Symptom Check List SCL-90R demonstrated good discrimination between patients with affective and anxiety disorders. However, the SCL-90R failed to discriminate patients with somatoform disorders from affective- and anxiety-disordered subjects. Therefore, the development of other psychometric scales is necessary for the evaluation of patients with somatoform disorders.

Adult↗
PubMed1992

Phosphorus-31 magnetic resonance metabolite imaging in the human body.

This work examines the feasibility of three-dimensional phosphorus-31 magnetic resonance spectroscopic imaging (31P MRSI) of metabolites in the human body using nonselective excitation with a single large circular surface coil for transmitting and receiving. The potential and limitations of this approach to clinical imaging are demonstrated on four selected examples: normal liver and heart, hematoma in the calf, and lymphoma in the groin. The obtained metabolite images showed anatomical detail and allowed differentiation of body organs and pathologic tissue from adjacent tissue. Three-dimensionally localized 31P spectra were reconstructed from nominal volumes of 4 to 15 cm3. These spectra showed characteristic resonances and metabolite intensity ratios for the tissue of origin demonstrating good three-dimensional localization. We conclude that surface coil 31P MRSI of body organs to map metabolite distributions is practically feasible with this approach, but due to experimental limitations, clinical utility requires technical improvements.

Heart↗
PubMed1992

Effect of supranormal coronary blood flow on energy metabolism and systolic function of porcine left ventricle.

OBJECTIVE: The goal was to determine if supranormal coronary blood flow increases myocardial oxygen consumption, high energy phosphate levels, and systolic function in the in situ autoperfused heart. METHODS: Thirteen anaesthetised open chest pigs with an intact, autoperfused coronary circulation, weight 30-40 kg, were studied. Measurements were made under basal conditions and during regional hyperperfusion of the anterior left ventricle produced by intracoronary infusion of adenosine (mean dose 3.3 mumol.min-1). Doppler coronary blood flow velocity in the anterior descending coronary artery, arterial and anterior interventricular venous blood oxygen content, high energy phosphates (by transmurally localised 31P NMR), and myocardial wall thickening (by sonomicrometry) were measured. RESULTS: With adenosine, coronary flow was increased to 355(SEM 59)% of control. Supranormal coronary flow produced no significant changes in anterior left ventricular oxygen consumption [99(12)% of control]. 31P NMR spectroscopy revealed no significant changes in the peak intensities of phosphocreatine or ATP in either the subendocardium or subepicardium (90-97% of control). Systolic anterior left ventricular wall thickening also did not change [107(13)% of control]. CONCLUSIONS: Supranormal coronary flow does not augment myocardial oxygen consumption, high energy phosphates, or systolic function in the in situ autoperfused heart. Myocardial oxygen delivery does not limit oxidative metabolism under normal conditions.

Adenosine↗
PubMed1992

Evaluation of myocardial perfusion abnormalities with gadolinium-enhanced snapshot MR imaging in humans. Work in progress.

To determine whether myocardial perfusion abnormalities could be detected in patients with coronary artery disease by means of contrast material-enhanced magnetic resonance (MR) images, a snapshot imaging technique was used in six patients with coronary artery disease and four healthy subjects in conjunction with pharmacologic stress (dipyridamole infusion) and bolus injection of gadopentetate dimeglumine. MR images from all patients and healthy subjects were quantitatively analyzed to define spatial changes in signal intensity after administration of dipyridamole and gadopentetate dimeglumine. The resultant findings were compared with findings on thallium-201 scintigrams obtained after administration of dipyridamole and on coronary arteriograms in all patients. Nine myocardial regions supplied by stenosed arteries showed diminished levels of signal intensity after infusion of the contrast agent compared with those of normally perfused regions. These findings were in agreement with those obtained with T1-201 scintigraphy (in eight of nine regions) and arteriography. Thus, contrast-enhanced high-speed MR imaging with use of dipyridamole enabled detection of regional perfusion abnormalities in humans.

Adult↗
PubMed1992

Response of high-energy phosphates and lactate release during prolonged regional ischemia in vivo.

BACKGROUND: The functional impairment of persistently ischemic, or "hibernating," myocardium may serve to maintain myocardial cell viability through a reduction of energy requirements. Although previous studies have, in a variety of experimental models, independently shown variable responses in lactate metabolism and intracellular phosphates during prolonged ischemia, the responses of these metabolites under identical flow conditions have not been adequately described. METHODS AND RESULTS: To examine the responses of high-energy phosphates and lactate metabolism to prolonged ischemia induced by partial coronary artery stenosis, 12 open-chest pigs were studied using 31P nuclear magnetic resonance spectroscopy. Concurrent measurements of blood flow, segment shortening, high-energy phosphates, and lactate release (in nine animals) were made during 2 hours of regional ischemia. Subendocardial blood flow and segment shortening were persistently depressed during ischemia, with parallel reductions in ATP, phosphocreatine (PCr), and the ratio of phosphocreatine to inorganic phosphate (PCr/Pi). Pi was persistently elevated during the ischemic period. In contrast, lactate release increased significantly from 0.23 +/- 0.04 to 1.34 +/- 0.28 mumol/ml after 15 minutes of ischemia (p less than 0.05) but then decreased to 0.73 +/- 0.17 mumol/ml at 2 hours (p less than 0.05 versus 15 minutes, p = NS versus control). Similarly, pH increased significantly from a nadir of 6.82 +/- 0.07 at 30 minutes of ischemia to 6.98 +/- 0.05 at 2 hours. CONCLUSIONS: Changes in high-energy phosphates parallel changes in blood flow and function during prolonged ischemia, whereas there is a partial amelioration in lactate production and acidosis. These data support the concept that reduction of myocardial energy requirements during prolonged flow reduction results in signs of reduced ischemia.

Animals↗
PubMed1991

31P MRS of myocardial inorganic phosphate using radiofrequency gradient echoes.

Determination of the chemical shift and integral of the myocardial intracellular inorganic phosphate (Pi) resonance by 31P magnetic resonance spectroscopy (MRS) is often precluded due to a large overlapping signal from 2,3-diphosphoglycerate (2,3-DPG) from chamber and myocardial blood. This report demonstrates the use of radiofrequency (RF) magnetic field gradient echoes (RFGE) to eliminate signals from 2,3-DPG in flowing blood, while retaining signals from intracellular myocardial Pi, ATP, and phosphocreatine (PCr). The ECG-triggered 31P spectra were acquired from the myocardium of open chest pigs using a Philips Gyroscan 2-T magnetic resonance spectrometer. A 2.5-cm-diameter surface coil attached to the myocardium was used to provide the RF gradient as well as for excitation and detection of signals. Optimal performance of the RFGE pulse sequence was obtained when the RF gradient pulses were centered at peak diastole or peak systole. Under these conditions, 2,3-DPG signals were completely suppressed, and sensitivity was usually sufficient to allow detection of a well-resolved Pi signal. Myocardial pH determined from RFGE experiments was 7.16 +/- 0.10, and the ratio of the integrals of the Pi and ATP resonances (Pi/ATP) was 0.24. The mean signal-to-noise ratio (S/N) for PCr in control spectra acquired in 4 min was 19/1, while the mean S/N for PCr in RFGE-edited spectra acquired in 15 min was 11/1, demonstrating that the present implementation of the RFGE method results in significant loss in sensitivity. These experiments demonstrate that RFGE-editing allows accurate determination of the chemical shift and integral of the Pi resonance in blood-perfused myocardium in situ.

2,3-Diphosphoglycerate↗
PubMed1991

DNA replication of wheat dwarf virus, a geminivirus, requires two cis-acting signals.

Replication of the single-stranded DNA genome of wheat dwarf virus (WDV) leads to the accumulation of covalently closed double-stranded DNA of genome length in infected cells. By studying the replication properties of a naturally occurring deletion mutant of WDV isolated from infected plants and of deletion mutants constructed in vitro, we have defined cis-acting regions required for viral DNA replication. The results show that two distinct regions are required in cis to yield the normal replicative forms of WDV-DNA.

Blotting, Southern↗