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Biomedical subjects

W Hemmer

Publications and source records attributed to W Hemmer.

At least 55 records · Page 3Linked to original sources
PubMed1998

Expression of intercellular adhesion molecule-1 in human coronary endothelial and smooth muscle cells after stimulation with tumor necrosis factor-alpha.

BACKGROUND: The intercellular adhesion molecule-1 (ICAM-1) is one of several human cell adhesion molecules that play a critical role in the early stages of postangioplasty restenosis. In this study, the in-vitro expression of ICAM-1 in human coronary endothelial cells and human coronary smooth muscle cells (SMC) after stimulation with tumor necrosis factor-alpha (TNF-alpha) was investigated. METHODS AND RESULTS: SMC were isolated from the media of normal human coronary arteries (n = 26) up to 10 h post mortem (HCMSMC) and from human atherosclerotic coronary arteries (HCPSMC) that were extracted by thrombendarterectomy (n = 25). Endothelial cells of human coronary arteries (HCAEC) were purchased from Clonetics (Cell System, Remagen, Germany), and endothelial cells from human umbilical cord veins (HUVEC) were isolated after vaginal delivery. For investigations of the effect of TNF-alpha (2.5, 5, 10, and 20 ng/ml) on the proliferative activity of HUVEC, HCAEC, HCPSMC, and HCMSMC, serum-free media was used. After 24 h cell number and cell size distribution were measured in a cell analyzer system. The proliferation of HCPSMC and HCMSMC was increased by TNF-alpha; however, significant differences compared with controls were not reached. The proliferation of HUVEC and HCAEC was significantly reduced by TNF-alpha. For investigations of the effect of TNF-alpha (2.5, 5, 10, and 20 ng/ml) on the surface expression of ICAM-1, monoclonal anti-ICAM-1 antibodies (84H10) were used. The expression of ICAM-1 was analyzed using an immunofluorescence microscope. For flow cytometry analysis, 5 x 10(3) cells (100% gated) were analyzed using a fluorescence-activated cell sorter. In control cultures with no stimulation, the expression of ICAM-1 was positive in HCAEC, HCPSMC, HCMSMC, and HUVEC. TNF-alpha stimulated the expression of ICAM-1 in a time- and dose-dependent manner. After maximal stimulation with TNF-alpha (20 ng/ml for 24 h), the expression of ICAM-1 was stronger in HCMSMC than in HCPSMC. CONCLUSIONS: These results suggest that the cytokine TNF-alpha regulates the expression of ICAM-1 in both human coronary endothelial cells and SMC, and could therefore play an important role in the pathophysiology of inflammatory and immune processes in restenosis after angioplasty.

Cell Division↗
PubMed1998

Identification of allergens in oilseed rape (Brassica napus) pollen.

BACKGROUND: Pollen from oilseed rape (OSR), Brassica napus, an increasingly cultivated oilplant from the Brassicaceae, has been recognized as a potential cause of allergic sensitization. Allergens have been hardly investigated. METHODS: We characterized IgE binding proteins in OSR pollen by immunoblot, immunoblot inhibition and specific monoclonal antibodies using sera from 89 patients sensitized to OSR. RESULTS: Two low-molecular-weight allergens of 6/8 kD and 14 kD as well as a high molecular-weight cluster (27-69 kD) comprising six cross-reactive peptides could be identified. The three allergens were recognized by 50, 34 and 80% of patients, respectively. Immunoblot IgE binding to OSR could be totally inhibited by rye pollen and moderately by birch pollen (6/8 and 14 kD) while mugwort had little effect. An anti-profilin-specific monoclonal antibody bound specifically to a 14-kD protein in OSR. Binding to the 6/8-kD rape allergen could be effectively inhibited by rAln g 2, a calcium-binding protein from alder. Periodate treatment led to a significant reduction in IgE binding to the 27 to 69-kD OSR allergens indicating that carbohydrate determinants are involved in IgE binding. OSR proteins were capable to quench IgE binding to timothy grass pollen proteins of >/=60 kD suggesting that grass pollen group 4 allergens cross-react with the 27 to 69-kD cluster in OSR. CONCLUSIONS: The data demonstrate that OSR pollen is allergenic and indicate that the identified allergens represent cross-reacting homologues of well-known pollen allergens, i.e. calcium-binding proteins, profilins, and high-molecular-weight glycoproteins. Via cross-reactivity, exposure to OSR pollen may be a prolonging and aggravating factor in underlying birch and grass pollen allergy.

Allergens↗
PubMed1997

Role of the glycine triad in the ATP-binding site of cAMP-dependent protein kinase.

A glycine-rich loop in the ATP-binding site is one of the most highly conserved sequence motifs in protein kinases. Each conserved glycine (Gly-50, Gly-52, and Gly-55) in the catalytic (C) subunit of cAMP-dependent protein kinase (cAPK) was replaced with Ser and/or Ala. Active mutant proteins were expressed in Escherichia coli, purified to apparent homogeneity, separated into phosphoisoforms, and characterized. Replacing Gly-55 had minimal effects on steady-state kinetic parameters, whereas replacement of either Gly-50 or Gly-52 had major effects on both Km and kcat values consistent with the prediction of the importance of the tip of the glycine-rich loop for catalysis. Substitution of Gly-50 caused a 5-8-fold reduction in Km (ATP), an 8-12-fold increase in Km (peptide), and a 3-5-fold drop in kcat. The Km (ATP) and Km (peptide) values of C(G52S) were increased 8- and 18-fold, respectively, and the kcat was decreased 6-fold. In contrast to catalytic efficiency, the ATPase rates of C(G50S) and C(G52S) were increased by more than an order of magnitude. The thermostability of each mutant was slightly increased. Unphosphorylated C(G52S) was characterized as well as several isoforms phosphorylated at a single site, Ser-338. All of these phosphorylation-defective mutants displayed a substantial decrease in both enzymatic activity and thermal stability that correlated with the missing phosphate at Thr-197. These results are correlated with the crystal structure, models of the respective mutant proteins, and conservation of the Glys within the protein kinase family.

Adenosine Triphosphatases↗
PubMed1997

Recombinant strategies for rapid purification of catalytic subunits of cAMP-dependent protein kinase.

Knowledge of the crystal structure of the catalytic subunit (C) of cAMP-dependent protein kinase provided for the first time a molecular basis for probing function by site-directed mutagenesis. The purification of mutant C-subunits, however, presented new and unanticipated challenges due to instability, insolubility, and underphosphorylation of the altered proteins. To overcome these barriers, a rapid and efficient method for purifying recombinantly expressed C-subunits was developed. Purification to near homogeneity is achieved in less than 5 h. The procedure is based on colysis of bacteria that overexpress the C-subunit with bacteria that overexpress a poly-His-tagged mutant of the type II regulatory subunit H6RII (R213K). This mutant R-subunit with an altered cAMP binding site A forms holoenzyme rapidly in bacterial extracts, and the Ka (cAMP) for the resulting holoenzyme, 27-37 microM, is nearly 50-fold increased compared to holoenzyme formed with wild-type RII. Thus, after batchwise immobilizing the holoenzyme on Ni(2+)-resin, the free C-subunit can be directly eluted batchwise with high concentrations of cAMP. The method is described for the purification of wild-type C, with yields of approximately 5 mg/liter. In addition, a mutant subunit, C[G52S], which is defective in ATP binding and could not be isolated using previously described methods, was purified with equal efficiency.

Adenosine Triphosphate↗
PubMed1997

Ultrastructural alterations in the right and left ventricular myocardium following multiple low energy endocardial countershocks in anesthetized dogs.

Both high energy transthoracic and direct epicardial defibrillation can result in RV and LV myocardial damage, but little is known about the damage due to defibrillation using an endocardial RV electrode. Furthermore, disturbances in postdefibrillation oxidative metabolism have been reported and may be caused by primary injury of mitochondrial integrity and function, but information about ultrastructural mitochondrial alterations is rare. We therefore studied, in 13 fox hounds, RV and LV ultrastructural alterations following multiple low energy endocardial countershocks. Using an ICD and an endocardial defibrillation system a median of 54 (43-74) countershocks with a cumulative energy of 1,558 J (844-2,141 J) was delivered. After termination of countershocks, RV and LV myocardium was examined by electron microscopy. In both ventricles, severe myocardial alterations were found, including swollen mitochondria, disruption of mitochondrial crests, and loss of integrity of the mitochondrial inner and outer membranes. At the first time a semiquantitative score, originally developed for postischemic injury, was successfully used to grade the postcountershock mitochondrial alteration, which showed a more pronounced damage in the RV (2.69 +/- 0.22 points) compared to the LV (2.18 +/- 0.22 P = 0.021). We conclude that even the use of endocardial lead systems with low energy countershocks may lead to severe mitochondrial damage, especially in the RV.

Animals↗
PubMed1997

Diagnosis of trichothiodystrophy in 2 siblings.

Trichothiodystrophy (TTD) is a rare autosomal recessively inherited disorder which is characterized by sparse and brittle hair with low cystine content. It is often associated with physical and mental retardation. We report 2 cases of TTD in 2 sibs who were born to related parents. The children showed clinical features typical of TTD and in addition other symptoms such as epilepsy, ataxia, spasticity, strabismus, atopic dermatitis, dysarthria and hyperextensible fingerjoints. The sulfur content of hair was reduced to about 50% of normal values and scanning electron microscopy of hair showed trichorrhexis nodosa, trichoschisis, missing cuticle scales with weathering of hair shafts. Under polarizing microscopy an alternating dark and bright banding was found. The present cases show that the correct diagnosis of TTD in practice can be impeded for many years because of the heterogeneous clinical appearance and that the determination of the sulfur content in hair is a simple but indispensable method.

Ataxia↗
PubMed1997

Sex steroids in cardiopulmonary bypass.

BACKGROUND: Sex steroids have immunomodulatory effects. No data exist on alterations of sex steroids during cardiopulmonary bypass (CPB). Cardiac surgery with CPB releases an immunological response with complement and cytokine activation. RESULTS: Plasma estradiol and progesterone levels before and immediately after CPB were measured in 11 patients. During CPB, mean estradiol levels decreased from 29 to 15 pg/ml and progesterone levels rose from 0.13 to 0.90 ng/ml. These changes were statistically significant. CONCLUSIONS: These are the first preliminary results evaluating plasma levels of sex steroids during CPB. Whether alterations in estradiol and progesterone levels influence complement and cytokine activation during bypass or vice versa is currently being investigated.

Journal Article↗
PubMed1997

Median fibrillation frequency in cardiac surgery: influence of temperature and guide to countershock therapy.

OBJECTIVE: This study was designed (1) to investigate the effects of normothermic and hypothermic perfusion on the median frequency of the fibrillating myocardium, and (2) to elucidate whether frequency-guided countershock therapy improves countershock success during the reperfusion phase of cardiac surgery. DESIGN: Prospective, randomized study. SETTING: University hospital cardiac surgery room. PATIENTS: Thirty patients (first part of the study) and 38 patients (second part of the study) scheduled for elective coronary artery bypass surgery. METHODS AND RESULTS: During cardiopulmonary bypass, ventricular fibrillation (VF) was induced at a core body temperature of 34.1+/-0.2 degrees C (normothermia) (n=15) or at a core body temperature of 29.8+/-0.2 degrees C (hypothermia) (n=15). Using fast Fourier transformation of the ECG signal, median fibrillation frequency was recorded continuously for a period of 120 s. At the end of surgery, countershock was performed as soon as VF was recognized on the ECG monitor (X Hz group; n=19) or countershock was not performed until median fibrillation frequency had increased to the threshold of at least 5 Hz (5 Hz group; n=19). Median fibrillation frequency in the normothermic fibrillation group was statistically higher than in the hypothermic group. In the X Hz and 5 Hz countershock group, median fibrillation frequency before the first countershock attempt was 3.6+/-0.2 Hz and 5.4+/-0.1 Hz (p<0.0001), respectively. In the X Hz group, six countershocks resulted in supraventricular rhythm, 10 in VF, two in electromechanical dissociation, and one in asystole. In the 5 Hz group, 16 countershocks resulted in supraventricular rhythm, two in VF, and one in asystole (p=0.008). CONCLUSIONS: During normothermia, median fibrillation frequency is significantly higher than during hypothermic perfusion conditions. During the reperfusion phase of cardiac surgery, countershock success rate is significantly higher when a threshold of at least 5 Hz had been reached before the first countershock attempt.

Coronary Artery Bypass↗
PubMed1997

Oilseed rape pollen is a potentially relevant allergen.

Oilseed rape (Brassica napus) (OSR) is a partly wind-pollinated crop which has been increasingly cultivated both in Europe and overseas. Allergological data about OSR is scarce and controversial. We evaluated the frequency of sensitization to OSR pollen by skin prick test and RAST over a period of 1 yr. Airborne OSR pollen load and the agricultural role of this crop were analysed. Furthermore, six patients were investigated by immunoblot. In 4468 patients with suspect inhalant allergy investigated between June 1994 and May 1995, routine skin prick testing revealed OSR sensitivity in 7.1% of pollen-allergic patients. In all, monovalent sensitization was detected in nine patients. Routine pollen counts showed daily maxima not exceeding 50 grains/m3/24 h, but airborne OSR pollen has continuously increased during the last decade correlating with the increasing acreage. Characterization of OSR allergens by immunoblot revealed major allergens of 6/8 kD, 12/14 kD and in the high molecular weight range at 33, 42, 51, 58/61 and 70 kD. Some OSR proteins may cross-react with birch pollen allergens. In summary, the results suggest that OSR pollen is a moderate but true source of allergy and may sensitize despite low pollen exposure.

Allergens↗
PubMed1997

Shed mediastinal blood in 500 elective cardiac surgery patients: is there enough for retransfusion routinely?

OBJECTIVE: Since an increased use of several blood salvaging measures has contributed to a reduction in perioperative blood loss and the requirement for banked blood in recent years, the aim of this study was to establish current postoperative drainage losses in order to evaluate whether homologous retransfusion may be a useful measure to reduce autologous transfusion in elective cardiac surgery. DESIGN/SETTING: This prospective clinical investigation was performed at a University Intensive Care Unit during the first six hours following cardiac surgery. PATIENTS: 373 men and 127 women undergoing elective cardiac surgery were investigated. MEASURES: The amount of shed blood was measured four and six hours postoperatively. RESULTS: The average blood loss was higher in men than in women both in all operations as a whole (men, four hours: 223+/-73 ml, six hours 270+/-95 ml; women, four hours: 156+/-25 ml, six hours 195+/-22 ml), in valve replacement (men, four hours: 299+/-87 ml, six hours 350+/-101 ml; women, four hours: 187+/-30 ml, six hours: 219+/-31 ml) and in coronary artery bypass grafting (men, four hours: 197+/-69 ml, six hours: 242+/-83 ml; women, four hours: 128+/-15 ml, six hours: 173+/-18 ml). A blood loss of 400 ml was exceeded in 13% of men after valve replacement four and six hours postoperatively. In all other groups, less than 8% of patients had a loss of more than 400 ml both after four and after six hours. CONCLUSIONS: Postoperative drainage losses in elective cardiac surgery patients are small and a measurable advantage from retransfusion seems to be unlikely. We therefore endorse the routine use of shed mediastinal blood retransfusion in these patients.

Blood Loss, Surgical↗
PubMed1996

Allergic contact dermatitis to artificial fingernails prepared from UV light-cured acrylates.

BACKGROUND: Contact dermatitis from artificial nails made from self-curing acrylic resins is occasionally reported. Recently, UV light-cured products introducing new acrylics have become available. OBJECTIVE: Our purpose was to identify relevant allergens in commercial light-curing products by patch tests and to evaluate the efficacy of "hypoallergenic" products by inclusion into the test series. METHODS: Patients wearing photobonded acrylic nails who had perionychial and subonychial eczema were patch tested with an acrylate battery and "hypoallergenic" commercial products. RESULTS: Triethyleneglycol dimethacrylate, hydroxyfunctional methacrylates, and (meth)-acrylated urethanes proved to be relevant allergens in photobonded nail preparations. Methacrylated epoxy resin sensitization was not observed. All "hypoallergenic" products provoked positive reactions. CONCLUSION: The omission of irritant methacrylic acid in UV-curable products does not reduce the high sensitizing potential of new acrylates. In contrast to the manufacturers' declarations, all "hypoallergenic" products continue to include acrylate functional monomers and therefore continue to cause allergic sensitization.

Acrylates↗
PubMed1996

Formaldehyde and phenol exposure during an anatomy dissection course: a possible source of IgE-mediated sensitization?

The sensitizing potency of formaldehyde and phenol exposure during 4 weeks of an anatomy dissection course was assessed in 45 medical students. Specific IgE against formaldehyde by RAST and by ELISA and specific IgE against phenol by ELISA were assessed before and after the course. At the start of the course, symptoms, type I allergy, respiratory diseases, and smoking habits were noted. At the end of the course, only symptoms experienced during the dissection lessons were assessed. Indoor formaldehyde levels were measured continuously. The mean indoor formaldehyde level was 0.124 +/- 0.05 ppm, with a minimum of 0.059 ppm and a maximum of 0.219 ppm. Specific IgE against formaldehyde or phenol was found in none of the subjects at the beginning of the course, and no student showed specific IgE against formaldehyde or phenol after the course. Assessment of primarily irritant symptoms during the lesson revealed itch and paraesthesia of hands in 33/45 students (P < 0.00005), headache in 15/45 students, burning eyes in 13/45 students (P < 0.02), dizziness in 8/45 students (P < 0.008), sneezing in 4/45 students, epistaxis in 2/45 students, and shortness of breath in 1/45 students. According to our data, 1-month exposure to formaldehyde and phenol during an anatomy dissection course does not induce specific IgE against formaldehyde or phenol.

Adolescent↗