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Biomedical subjects

H Wolf

Publications and source records attributed to H Wolf.

At least 397 records · Page 22Linked to original sources
PubMed1990

Non-radioactive labeling and detection of nucleic acids. III. Applications of the digoxigenin system.

The digoxigenin-based non-radioactive DNA labeling and detection system was applied in various hybridization protocols using digoxigenin-labeled probes obtained by enzymatic incorporation of Dig-[11]-dUTP. In genomic blots single-copy genes (human tissue-type plasminogen activator, constant part of immunoglobulin kappa light chain) can be detected with only 0.5 to 5 micrograms human DNA depending on the type of probe and the length of the hybridizing region. Due to its high sensitivity and specificity, the digoxigenin system is also appropriate for colony-, plaque-, and in situ hybridizations with metaphase chromosome spreads and fixed cells. Especially in the latter applications it is of great advantage, that with the digoxigenin system any significant background or unspecific side reactions with biological materials are avoided.

Blotting, Southern↗
PubMed1990

Monoclonal antibodies against the free subunits of human chorionic gonadotrophin.

Discordant results on body fluid levels of human chorionic gonadotrophin (hCG) free alpha- and beta-subunits under physiological and pathophysiological conditions, prompted us to raise a total of 260 monoclonal antibodies (MCA) against free hCG-alpha, free hCG-beta, holo-hCG, human follicle-stimulating hormone and bovine luteinizing hormone; 153 MCA recognizing the human alpha-subunit and 28 reacting with hCG-beta were extensively analysed for their intra- and inter-species cross-reactivity with homologous hormones, and for the compatibility of epitopes recognized by them. The immunological topography of free hCG-alpha and free hCG-beta was resolved by these MCA, and epitope maps were designed. Six antigenic determinants on the free alpha-chain (alpha 1-alpha 6), clustered in three spatially distinct domains, and seven epitopes on the surface of free hCG-beta (beta 1-beta 7), could be distinguished. Strikingly, three alpha-chain epitopes (alpha 4, alpha 5 and alpha 6) were shared between various species, which is in contradiction to the concept of immunological species-specificity of alpha-subunits. Three determinants were found to be present only on the free subunits but not on holo-hCG (alpha 6, beta 6 and beta 7), and only two determinants (beta 1 and beta 7) were hormone-specific for hCG. Based on this information, an immunoenzymometric assay for the free alpha-subunit of human glycoprotein hormones was established, with a sensitivity of 1.3 pg/well and a cross-reactivity with holo-hCG of less than 0.005%.(ABSTRACT TRUNCATED AT 250 WORDS)

Antibodies, Monoclonal↗
PubMed1990

Investigation of blood-biomaterial interaction by means of a new quantitative dynamic measuring principle.

In this study a new dynamic measuring chamber for flat biomaterials allowing the estimation of cell adhesion on test surfaces by measuring the cell lost from surface contacting test blood under defined rheological conditions is introduced. This was achieved by constructing a test chamber permitting the contact of small amount of blood with a large geometrical test surface. The construction consists of a spiral-shaped flow channel of 0.3 cm width, 0.02 cm height and 78 cm length. The applicability of the new method was demonstrated by evaluation of platelet adhesion on siliconized glass, fibrinogen coated glass, cuprophane and polyacrylnitrile.

Biocompatible Materials↗
PubMed1990

[Acute phase reaction in acute viral hepatitis].

We investigated serum concentrations of acute-phase-proteins (alpha 1-antitrypsin (alpha 1-AT), alpha 1-acid glycoprotein (alpha 1-sGP) and C-reactive protein (CRP)), and of complement components C3 and C4 during the course of acute hepatitis A and hepatitis B and correlated it to age and sex of the patients. Whereas a small increase of all three acute-phase-proteins was seen during acute hepatitis A, alpha 1-AT and CRP, but not alpha 1-aGP were found to be elevated during hepatitis B. A moderate elevation of C3 and C4 serum levels, more pronounced in hepatitis B than in hepatitis A, was seen followed by a slow return to normal. A systemic acute phase response seems not to be present during acute viral hepatitis in spite of the pronounced local inflammatory changes in liver tissue.

Acute-Phase Proteins↗
PubMed1990

[Effectiveness of various combination therapies in patients with coronary heart disease].

There is only a limited number of studies available comparing the effectiveness of various combinations of anti-ischemic and antianginal substances in the same patients with coronary artery disease and stable angina pectoris and even these are restricted to either only a few drugs or a single point in time for testing. Accordingly, this study was undertaken to determine to what extent the combination of two or three drugs with different anti-ischemic mechanisms of action such as the long-acting form of the beta-blocker metoprolol and isosorbide dinitrate (ISDN) in sustained-release form as well as the calcium channel blockers nisoldipine and diltiazem in sustained-release form, which previously have not been tested in combination, are capable of enhancing effectiveness and prolonging duration of action. In a double-blind, randomized, crossover study in eleven patients with documented coronary artery disease and stable angina pectoris the effects of monotherapy with 200 mg metoprolol in long-acting form were compared with those of combined treatment with 120 mg ISDN sustained-release or 10 mg nisoldipine or 120 mg diltiazem sustained-release as well as ISDN and nisoldipine and finally, ISDN and diltiazem by means of an intraindividual analysis. For assessment of anti-ischemic and antianginal effects, symptom-limited exercise testing was carried out before as well as three, eight, twelve and 24 hours after medication. The parameters analyzed were ST-segment depression at the highest comparable workload, ischemia-free and symptom-free exercise capacity (one minute prior to ST-segment depression of 1 mm or onset of angina pectoris) as well as the systolic blood pressure--heart rate product at the highest comparable workload and at the highest ischemia-free workload, that is one minute prior to an ischemic reaction of 1 mm. Based on the ST-segment depression, all combinations of two drugs (metoprolol and ISDN at three hours; metoprolol and diltiazem at eight hours) led to a significant or at least relative increase of effectiveness. On comparison of the various double combinations, those with nisoldipine showed an early dissipation of action which, twelve hours after administration, was significantly less marked than those with diltiazem. Of the two tested triple combinations, metoprolol, ISDN and diltiazem was either significantly more effective than the various double combinations (metoprolol and ISDN or metoprolol and nisoldipine, both at eight and twelve hours; metoprolol and diltiazem, twelve hours) or relatively more effective and showed clear prolongation of the effects in excess of twelve hours.(ABSTRACT TRUNCATED AT 400 WORDS)

Adrenergic beta-Antagonists↗
PubMed1990

[Differences in free amino acid serum levels in premature neonates fed Feminar supplemented with whey proteins].

Free amino acids were examined in plasma of premature infants fed adapted dried milk (15 and 22 infants resp.) and breastfed infants (20). The dried milk was Feminar (casein: whey protein 40:60) where the whey (serum) proteins were prepared either by thermal denaturation (F-TD) or by ultrafiltration (F-U). The concentration of different amino acids in infants fed F-TD were closer to those recorded in breastfed infants, as compared with F-U where greater differences (higher values) were recorded.

Amino Acids↗
PubMed1990

Carbohydrate components of human immunodeficiency viruses type I and II.

The terminal carbohydrate residues of HIV I and II were detected by ferritin labeled lectins in electron microscopy. Different cell lines, which were infected with HIV I and II, expressed different terminal carbohydrate residues, which could also be detected on the viral envelope by electronmicroscopy. Especially N-Acetylgalactosamine residues were detected by Vicia villosa agglutinin only on Jurkat cells. This may have functional implications, since this lectin recognizes contrasuppressor T cells.

Carbohydrate Metabolism↗
PubMed1990

[Efficacy and side effects of differential calcium and phosphate administration in prevention of osteopenia in premature infants].

The prevention of osteopenia and frequency of renal and intestinal side effects of mineral supplementation was studied in 24 preterm infants with birth weight under 1,500 g, prospectively (gestational age 26-34 weeks). Calcium intake varied from 2.5 vs. 3.75 vs. 5 mmol/kg/day, phosphate was offered in dose of 2.5 mmol/kg/day. At the expected birth date 40% of infants with low calcium dose showed an activity of serum alkaline phosphatase greater than five times the maximum adult normal value which is defined as a reliable marker; for osteopenia no infant with medium or high calcium intake reached this critical value (p = 0.03). Medium and high calcium doses resulted in an increased risk for hypercalcuria (25 vs. 50%) (p = 0.03). Half of these infants developed typical signs of nephrocalcinosis on ultrasound examination. No significant difference of fecal fat content was observed with increased calcium intake; but more episodes of abdominal distension occurred during the first days of high calcium supplementation (p = 0.03). We conclude, that a calcium intake of 3.75 mmol/kg/day in combination with phosphate 2.5 mmol/kg/day is sufficient for adequate bone mineralization on a low level of side effects. Calcium excretion in urine has to be observed for early diagnosis of nephrocalcinosis.

Alkaline Phosphatase↗
PubMed1990

[Treatment of symptomatic and asymptomatic myocardial ischemia using 120 mg delayed-action ISDN].

Most episodes of myocardial ischemia in patients with coronary artery disease are incurred asymptomatically during everyday physical activities. While the necessity for medical treatment of angina pectoris is clearly established, the indication for treatment of asymptomatic ischemia is based on prevention of structural myocardial damage or malignant arrhythmias and on the implication of improvement in prognosis. In this regard, however, no reliable data is available. Additionally, only relatively few controlled studies have been carried out to investigate the influence of medical treatment on the ischemic episodes. Moreover, on assessment of treatment with Holter monitoring, as opposed to standardized ergometric testing, the substantial spontaneous variability of the frequency of ischemic episodes from day to day must be taken into consideration. Accordingly, in 25 patients with documented coronary artery disease, using a double-blind, randomized, placebo-controlled protocol with two periods of 48 hours of Holter monitoring each, we analyzed the effects of 120 mg isosorbide dinitrate in sustained-release form on the frequency, duration and extent as well as the circadian variation of transient myocardial ischemia during everyday physical activities and differentiated these from the spontaneous day-to-day fluctuations. During the placebo phase, 277 episodes with ST-segment depression greater than 1 mm were detected, 81% of which were asymptomatic. During treatment with 120 mg isosorbide dinitrate in sustained-release form, the number of episodes was reduced significantly (p less than 0.05) to 119 (-57%) where the decrease in symptomatic and asymptomatic episodes of 54% and 58%, respectively, was comparable (Figure 1).(ABSTRACT TRUNCATED AT 250 WORDS)

Coronary Disease↗
PubMed1989

Stereologic estimates of nuclear volume in noninvasive bladder tumors (Ta) correlated with the recurrence pattern.

The mean nuclear volume of cells in bladder tumors was estimated using the principle for estimation of the volume of particles of arbitrary shape. The study included three groups of patients: one group of 10 patients with only one nonrecurrent, noninvasive bladder tumor (Ta), another group of 11 patients with recurrent noninvasive bladder tumors, and a third group of 14 patients with recurrent primary noninvasive bladder tumors who ultimately developed an invasive tumor. After standard fixation, embedding, sectioning, and hematoxylin-eosin staining an unbiased estimate of the mean volume of nuclei sampled with a chance proportional to their volume was calculated: nu v = pi/3.l3(0). Here lo is the length of the intercept through a test point hitting a nucleus measured in a random direction through the test point. In the primary tumor the mean, nuclear volume of the cells in tumors from patients with a single bladder tumor is small (141 micrograms 3 geometric mean) with only one of ten above 165 microns 3. The mean nuclear volume of the primary tumor cells in patients with recurrent noninvasive bladder tumors was slightly larger (195 microns 3 geometric mean) with five of 11 above 165 microns 3. The mean nuclear volume of the primary tumor cells in patients who ended up with invasive tumors was higher (245 microns 3 geometric mean) with 12 of 14 above 165 microns 3. A large mean nuclear volume of cells in noninvasive bladder tumors (Ta) indicates recurrence and an invasive potential. Considering that nearly all primary tumors were classified as Bergkvist Grade II tumors, the prognostic precision of absolute mean nuclear volume is noticeable. This simple and fast estimate of mean nuclear volume seems to provide objective data with high prognostic value in primary noninvasive bladder tumors (Ta).

Aged↗
PubMed1989

Hepatitis B virus surface antigen as a reporter of promoter activity.

The coding sequence for the hepatitis B virus surface antigen (HBsAg) was used as a new reporter gene for studies on eukaryotic promoter activity and upstream regulatory sequences. The data observed in transfection assays were comparable to results obtained with conventional chloramphenicol acetyltransferase (CAT) assays, as was demonstrated using various transcriptional regulation sequences. The expression of HBsAg as a reporter protein offered some advantages: (i) In transient expression assays, a time course of promoter activity depending on variable culture conditions could be monitored over a period of time, since the HBsAg was secreted into the culture supernatant. (ii) Evaluation of HBsAg from supernatant aliquots and quantification of the corresponding promoter activities could be performed easily, using the very sensitive and readily available diagnostic HBsAg kits. (iii) In contrast to the conventional CAT assay, the cells remained available for further tests, e.g., Western blot, immunofluorescence or transcript analysis. Characteristics of several Epstein-Barr virus (EBV) promoters, depending on the virus state of EBV-positive B-cells (latency, chemical induction, lytic superinfection, trans-activation), were assayed using the HBsAg reporter system.

Animals↗
PubMed1989

Binding of wheat and peanut lectins to human transitional cell carcinomas. Correlation with histopathologic grade, invasion, and DNA ploidy.

The binding of peanut (PNA) and wheat germ (WGA) lectins to tissue sections was examined in biopsy specimens from normal urothelium (ten patients) and from tumor tissue of noninvasive (17 patients) and invasive bladder (31 patients) carcinomas. The results were correlated to DNA content, histopathologic grade, and the presence or absence of invasion. Significant alterations in lectin binding associated with the development of cancer were found. A gradual loss of both PNA and WGA binding was found to correlate with higher grades of atypia (P less than 0.001). The loss of WGA binding was significantly correlated with both tumor aneuploidy (P less than 0.001) and the presence of invasion (P less than 0.05), whereas no significant correlation was found between loss of PNA binding and these variables. We concluded that the loss of WGA binding structures associated with bladder cancer shows a better correlation with known risk factors (aneuploidy and invasion) than the loss of PNA binding does.

Arachis↗
PubMed1989

Macrophage colony-stimulating factor (CSF-1) is expressed by spontaneously outgrown EBV-B cell lines and activated normal B lymphocytes.

Human B lymphocytes activated by mitogens or infected by Epstein Barr virus (EBV) have previously been shown to release colony-stimulating activity (CSA) supporting the growth of normal human bone marrow progenitors. We established five different human EBV-B cell lines spontaneously outgrown from nonmalignant peripheral blood cells and long-term bone marrow cultures. CSA derived from all of these lines induces the growth of murine macrophage colonies, whereas virtually no human bone marrow cell progenitors were stimulated. As observed in the tumor cell line MIA PaCa-2, a 4.3-kilobase (kb) transcript was detected in all cases using a human colony-stimulating factor (CSF)-1 probe. Expression of this transcript can be further stimulated within three hours upon addition of phorbol myristate acetate (PMA). The highly purified native protein exerting macrophage colony-stimulating activity (M-CSA) exhibits a molecular size of approximately 75 to 97 Kd in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The identity of EBV-B cell derived M-CSA with human urinary CSF-1 was confirmed by a complete neutralization of macrophage CSA by an antihuman urinary CSF-1 antiserum. Normal human B lymphocytes purified from tonsils or from mononuclear blood cells also express CSF-1 upon stimulation with Staphylococcus aureus Cowan I. No CSF-1 expression, however, could be detected in normal resting B lymphocytes or in the Burkitt lymphoma cell line RAJI.

Animals↗
PubMed1989

Blood group ABH-related antigens in normal and malignant bladder urothelium: possible structural basis for the deletion of type-2 chain ABH antigens in invasive carcinomas.

A complete panel of mouse monoclonal antibodies (MAbs) against Type-2 chain (GaI beta I-4GlcNAc-R) blood-group antigens (N-acetyl-lactosamine, Lex, H, Ley, A monofucosylated, Aley, repetitive A) was used in a detailed immunohistological study of the modulation of these carbohydrate antigens in transitional-cell carcinomas. The histological and cellular locations of these antigens were studied in 19 normal bladder biopsies and 53 transitional-cell carcinomas with as well as without neuraminidase treatment of tissue sections in order to uncover potential sialylated antigens. The antigen expression was correlated to individual A1A2BO, Lewis, and secretor status. Several alterations of blood group expression were found: (1) loss of A and H antigens with accumulation of Ley antigens; (2) loss of correlation between antigen expression and secretor status; (3) disruption of the orderly stratification of blood-group antigen expression in relation to cell layers; and (4) changes in subcellular location of antigen expression. The present data indicate that deletion of Type-2 chain ABH antigens in transitional-cell carcinomas is associated with alpha 1-3 fucosylation of the H antigen leading to accumulation of Ley antigens.

ABO Blood-Group System↗
PubMed1989

Carrier-bound synthetic peptides. Use as antigen in HIV-1 ELISA tests and in antiserum production.

Chemically synthesize carrier-bound peptides have been used as antigens in diagnostic test systems (ELISA) and for raising antipeptide-specific antisera. The method does not require prior cleavage of the peptides from the support used for the solid-phase synthesis. Using the same resin for both the synthesis and the subsequent applications it was possible to avoid expensive and time-consuming purification procedures and artificial recoupling to solid supports. A quick and specific ELISA-based diagnostic test system for HIV-specific antipeptide antibodies in human sera was established. In addition the carrier-bound peptides were shown to be potent antigens for raising antibodies in animals.

Animals↗
PubMed1989

Detection of hepatitis B virus in serum using amplification of viral DNA by means of the polymerase chain reaction.

A new assay was developed for the detection of hepatitis B virus (HBV) in human serum using amplification of a short viral DNA sequence by means of the polymerase chain reaction. As little as 0.4 fg viral DNA, corresponding to about 130 genome equivalents, per ml serum could be detected after the amplification procedure. This assay detected viral DNA in a number of patients with proven or suspected chronic HBV infection who were all negative for HBV DNA in the conventional hybridisation assay. We found HBV DNA in all of six HBeAg-positive and in three of eight HBeAg-negative HBsAg carriers, as well as in all of 11 patients with chronic liver disease with antibodies against the HBV core antigen (anti-HBc) as the sole marker for HBV infection, and in three of five apparently healthy individuals showing only anti HBc. Thus, this method is an important improvement for the diagnosis of persistent HBV infections, especially in patients where a definitive serological diagnosis is not possible.

Antibodies, Viral↗
PubMed1989

Modification of HLA expression as a possible factor in the pathogenesis of Burkitt's lymphoma.

Our data show that: Most BL cells express significantly less HLA class I and II than the corresponding LCLs. Lymphoblastoid cell lines have a much higher HLA class I and II density than normal peripheral blood B cells. There are qualitative differences in class I and II expression in most BL/LCL pairs. These qualitative differences are not caused by EBV infection. They might be due to a selective downregulation of HLA specifities by c-myc activation. We conclude that these modifications of HLA expression may well play a role in the reduced specific immune recognition of Burkitt's lymphoma cells.

Burkitt Lymphoma↗