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Biomedical subjects

E Wolf

Publications and source records attributed to E Wolf.

At least 19 recordsLinked to original sources

Enzyme and immunohistochemistry on undecalcified bone and bone marrow biopsies after embedding in plastic: a new embedding method for routine application.

A simplified method of low temperature methyl and butyl methacrylate embedding (up -20 degrees to -15 degrees C) is demonstrated using a proper redox system of benzoyl peroxide and aromatic amine. This method combines the morphological superiority of plastic-embedded bone tissue and bone marrow sections with the advantages of specific enzyme histochemical and immunochemical markers. The method permits good preservation of morphological details, the survival of antigenic determinants and the retention of enzyme activities. The specimens were fixed in 1.6% formaldehyde and 5% sucrose in 0.02 M phosphate buffer at pH 7.4, washed in 0.02 M phosphate buffer and 5% sucrose, dehydrated with acetone and impregnated with monomers of embedding medium. All these steps were carried out at +4 degrees C. The method presented is especially suitable for enzyme histological and immunohistological diagnosis of primary and secondary bone tumours, soft tissue tumours, as well as myelo- and lymphoproliferative disorders of bone marrow biopsies. Examples are demonstrated with mono- and polyclonal antibodies and reaction products of hydrolytic enzymes.

Biopsy

The GH-transgenic mouse as an experimental model for growth research: clinical and pathological studies.

The objectives and the methodology of mammalian gene transfer are discussed and findings in growth hormone (GH) transgenic mice are reported to illustrate the potential offered by genetically designed animal models for investigations in various areas of biomedical research. Transgenic mice expressing hybrid genes composed of either human or bovine GH coding sequences fused to the mouse metallothionein I promoter show high serum levels of heterologous GH, increased growth rates and final adult size, decreased life expectancy and a variety of pathological changes.

Animals

Definitions and characteristics of salt-sensitivity and resistance of blood pressure: should the diagnosis depend on diastolic blood pressure?

To elucidate the importance of diastolic blood pressure in the definition of salt-sensitive hypertension, we studied 54 male subjects, 36 of whom had untreated, mild essential hypertension. The subjects received a 120 mmol/d Na (as the chloride salt) diet for six days. Thereafter they received a 10 mmol/d Na diet for eight days followed by a 400 mmol/d Na diet for another 8 days. Blood pressure was measured hourly "around the clock" on the last day of each diet; the averaged systolic, diastolic and mean blood pressure values were compared. In 22 subjects diastolic blood pressure increased, when salt intake was increased from 10 to 400 mmol/d. In 18 of these 22 subjects systolic blood pressure increased as well. In 20 subjects, systolic blood pressure increased with salt loading while diastolic blood pressure decreased. In 13 subjects both systolic and diastolic blood pressure decreased with increased salt intake. We defined those subjects showing an increase in diastolic blood pressure as salt-sensitive. If mean blood pressure were used to define salt-sensitivity, 8 of our subjects would have been labeled as salt-sensitive who actually decreased their diastolic blood pressure with salt loading. We suggest that consideration of systolic and diastolic blood pressure responses gives better insight into identifying volume and resistance-related phenomena in salt-sensitive hypertension, than does the consideration of mean blood pressure alone. The definition of salt-sensitivity may require reassessment.

Adult

Sodium kinetics in salt-sensitive and salt-resistant normotensive and hypertensive subjects.

OBJECTIVE: To test the hypotheses that sodium kinetics are not affected by blood pressure, salt sensitivity, salt resistance or race, and that the kinetics of sodium balance are not a first-order process. DESIGN, PARTICIPANTS AND INTERVENTIONS: Two studies were conducted. In the first, 18 normotensive and 36 hypertensive men and women were given sodium at 120 mmol/day for 6 days, followed by 10 mmol/day for 8 days, then 400 mmol/day for 8 more days. Salt sensitivity was defined as an increase in diastolic blood pressure from the 10 to the 400 mmol/day intake. Salt resistance was defined as no increase, or a decrease in diastolic blood pressure with the increased sodium intake. In the second study, 12 white and 12 black normotensive men ingested sodium at 10, 200 or 400 mmol/day in random order, each for 7 days. All urine was collected in both protocols. SETTING: Metabolic ward at the University of Greifswald (Greifswald, Germany; study 1), and Clinical Research Center (Indiana University, Indianapolis, Indiana, USA; study 2). MAIN OUTCOME MEASURE: In addition to conventional statistics, a pharmacokinetic analysis was carried out to determine the elimination rate constant and half-life. RESULTS: In the Greifswald study, when the sodium intake was decreased, a longer half-life was determined for the salt-sensitive than the salt-resistant hypertensive subjects. The half-life for the normotensive salt-sensitive and salt-resistant subjects did not differ. When the sodium intake was decreased, a monoexponential equation fitted the data for all subjects; when the sodium intake was increased, only data for half the subjects could be fitted to the same equation. In the Indianapolis study, black race had a significant influence upon urinary sodium excretion. Furthermore, the half-life for sodium elimination was dependent upon sodium intake; namely, the greater the intake, the longer the elimination half-life. CONCLUSIONS: The time required to reach sodium balance may increase following salt-sensitive increases in blood pressure rather than precede them. Race influences the time required to achieve salt balance. Sodium kinetics are not a first-order process.

Adult

Accelerated growth and visceral lesions in transgenic mice expressing foreign genes of the growth hormone family: an overview.

Effects of growth hormone (GH) overproduction were studied in transgenic mice expressing murine metallothionein I-GH fusion genes. The most obvious consequence was the acceleration of growth, which led to substantial increases in body weight of up to more than twice that seen in controls. Growth of the internal organs was stimulated, with hepatomegaly and nephromegaly as the most prominent features. GH transgene expression was also reflected in increased skeletal growth which affected various bones to different extents. The mean life-span of human GH transgenic mice with serum levels of hGH ranging from 3 x 10(3) to 9 x 10(5) ng/ml was drastically reduced at 160 days in both sexes. Severe renal lesions were the primary cause of the decrease in life expectancy and were characterized by marked nephron atrophy, obsolescence of numerous glomeruli, and a massive cystic dilation of the tubules. Initial changes involved the glomeruli, which showed significant enlargement and sclerotic lesions. The liver exhibited a pronounced hepatocellularmegaly and progressive degenerative as well as hyperplastic changes. One-third of the hGH transgenic animals displayed myocardial fibrosis. Hepatocellular carcinoma was found in bovine GH transgenic mice older than 12 months. Our observations are compared with results of other investigators.

Animals

Color discrimination and accuracy of blood glucose self-monitoring in type I diabetic patients.

OBJECTIVE: To determine the importance of color discrimination ability regarding accuracy in the self-monitoring of blood glucose. RESEARCH DESIGN AND METHODS: Two hundred two insulin-dependent (type I) diabetic patients performed the Fansworth-Munsell 100-hue test and a second group of 159 type I diabetic patients performed a new Düsseldorf 26-hue test. Error scores in both tests were compared with error scores of patients' self-monitoring of blood glucose measurements with Chemstrip Haemoglucotest 20-800 strips. RESULTS: Color discrimination ability decreased with age, diabetes duration, and presence of retinopathy. It was independent of the degree of glycemia and accuracy of self-monitoring of blood glucose. CONCLUSIONS: Impaired color vision by itself is no reason to abandon self-monitoring of blood glucose with visually read strips.

Adult

Growth characteristics of metallothionein-human growth hormone transgenic mice as compared to mice selected for high eight-week body weight and unselected controls. I. Body weight gain and external body dimensions.

Body weight gain and external body dimensions of MT-hGH transgenic mice were compared with mice (NMRI) selected for high 8-week body weight (N8) and unselected controls derived from the NMRI strain (Pop). The growth curves from day 30 to 120 of transgenic mice exhibited a significantly steeper slope than those of male and female controls and of female N8 mice and did not show sex-related differences. The continuous pattern of GH secretion in transgenic mice is discussed as a possible reason for this phenomenon. Body weight gain of transgenic mice did not significantly exceed that of male N8 mice. None of the groups showed an obvious prolongation of the period of rapid daily weight gain. Maximum body weights of male and female transgenic mice were significantly higher than those of sex-matched controls but not of N8 mice. A drastic loss of body weight of about 25% of the maximum value was observed in the transgenic group prior to death. External body dimensions were largest in MT-hGH transgenic animals, followed by N8 mice and controls. In addition to these absolute measurements, values were related to the cube root of maximum body weight of the same animal. This is the first study that provides a comparative analysis of the effects of GH gene transfer and selection for body weight gain on body growth of mice derived from an outbred strain.

Animals

Growth characteristics of metallothionein-human growth hormone transgenic mice as compared to mice selected for high eight-week body weight and unselected controls. II. Skeleton.

Growth hormone and mechanical loading are known to be important factors influencing bone growth. We have measured proportions of the skull and the postcranial skeleton of metallothionein-human growth hormone (MT-hGH) transgenic mice expressing high levels of hGH in their serum, of NMRI mice being large as a result of selection for high 8-week body weight (N8), and of unselected controls (Pop) derived from the NMRI strain. Absolute bony dimensions of transgenic mice were as a rule significantly larger than those of controls, the differences ranging between 3% and 32% in males and from 6% to 28% in females. By contrast, the enlargement of skeletal dimensions of N8 mice did not exceed 10% and was restricted to distinct bones. When related to the cube root of maximum body weight of the same animal, bones of controls were as a rule larger than those of N8 and MT-hGH transgenic mice. A detailed analysis of bony dimensions of GH transgenic mice and of mice selected for high body weight was carried out to judge the effects of GH overexpression and mechanical loading due to increased body weight on bone growth. The fact that bones of transgenics were as a rule larger than those of selected mice in spite of both groups reaching similar maximum body weights, suggests that skeletal gigantism in MT-hGH transgenic mice can only in part be a result of increased body weight.

Animals

Expression of metallothionein-human growth hormone fusion genes in transgenic mice results in disproportionate skeletal gigantism.

Transgenic mice harbouring mouse metallothionein I-human growth hormone (MT-hGH) fusion genes were produced using the microinjection technique. The bones of adult MT-hGH transgenic mice, which continuously expressed high levels of hGH in their serum, and age-matched controls lacking detectable concentrations of hGH were measured microscopically. In addition to analyzing absolute skeletal dimensions, measurements were related to the cube root of the maximum body weight of the same animal. Absolute values obtained from transgenic mice were significantly higher than those obtained from controls for most of the defined measurements. However, the increase in skeletal dimensions was mostly not as pronounced as the increase in body weight and all bones were not affected to the same extent. There was no significant correlation between the serum GH concentration in individual mice and their degree of bony overgrowth. A disproportionate skeletal gigantism in MT-hGH transgenic mice may result from time differences in epiphyseal union of various bones of both sexes as well as differences in mechanical bone loading due to a drastically increased body weight. Individual concentrations of locally produced tissue insulin-like growth factor I (IGF I) might also play a role. Possible effects of these factors are discussed. The results presented in this study show that MT-hGH transgenic mice provide a powerful tool for the investigation of hormonal regulation of bone growth.

Amino Acid Sequence

[Unexpected transgene expression of a mammary-specific growth hormone gene construct in Bergmann glial cells of the mouse].

Pronuclear microinjection was used to produce transgenic mice harboring gene constructs, in which 110 base pairs (WAP1) or 2.4 kilobases (WAP2) of the 5' flanking sequences of the whey acidic protein (WAP) gene were fused to human growth hormone (hGH)-coding sequences. Female WAP-hGH transgenic mice expressed the transgenes in the mammary gland, the expression of the WAP2-hGH transgene mirroring that of the endogenous WAP gene. When other organs were examined, high level expression of hGH was observed in the brains of WAP2-hGH transgenic mice. Using in situ hybridization and immunohistochemistry, hGH expression from the transgene was seen to occur specifically in Bergmann glia cells. While normally neither WAP nor hGH is expressed in this type of cell, it appears that a combination of the regulatory region of the WAP gene and the hGH structural gene results in a novel tissue specificity in Bergmann glia cells.

Animals