Hans Dierck Waller

Lipid peroxidation in erythrocytes

Erythrocytes might be expected to be highly susceptible to peroxidation. Their membranes are rich in polyunsaturated fatty acids; they are continuously exposed to high concentrations of oxygen; and they contain a powerful transition metal catalyst. In fact, autoxidation is held in check in vivo by extremely efficient protective antioxidant mechanisms. These involve cellular enzymes such as superoxide dismutase and glutathione peroxidase, as well as vitamin E; but they mainly reflect effective structural compartmentalisation. This review surveys mechanisms which lead to red cell lipid autoxidation and the role of haemoglobin in these processes. The influence of haemoglobinopathies, of lipid composition and of abnormalities in antioxidant mechanisms induced by exogenous oxidant stress is also considered.

Sweet's syndrome associated with myelodysplasia : possible role of cytokines in the pathogenesis of the disease

Summary. The clinical course of a 56‐year‐old female patient with Sweets syndrome (SS) preceded by a myelodysplastic syndrome (MDS) is described. During the acute phase of the disease with high remittent fever, painful skin lesions and maximal leucocytosis IL‐6 and G‐CSF serum levels were extremely high, while TNF‐alpha was only slightly elevated and gamma‐interferon and IL1‐β were not increased. On clinical improvement IL‐6 serum levels rapidly fell, whereas G‐CSF values already slightly elevated before the manifestation of the disease slowly declined.

Polymerase chain reaction to evaluate antiviral therapy for cytomegalovirus disease

A sensitive and specific method to monitor suppression of cytomegalovirus (CMV) replication is essential in patients treated with ganciclovir after allogeneic bone-marrow transplantation. In this study, antiviral therapy of eighteen episodes of symptomatic CMV infection in 15 such patients were followed up clinically and by virus culture and polymerase chain reaction (PCR). Clinical improvement, culture, and PCR were assessed for their ability to predict the efficacy of ganciclovir therapy in each patient. In eleven successfully treated episodes of CMV disease (disappearance of symptoms and improvement in biochemical variables) clinical improvement was associated with an effective suppression of virus replication as shown by negative culture and PCR assays of blood and urine specimens obtained after antiviral therapy. 1 patient who did not improve clinically when receiving antiviral therapy remained both culture positive and PCR positive for CMV. 6 patients with early relapse of CMV disease or who died after an initial clinical improvement were PCR positive but culture negative after termination of therapy. Demonstration of CMV in blood and urine by PCR after stopping antiviral therapy (even when culture is negative) points to incomplete suppression of virus replication. The findings show that PCR is a better predictor of the efficacy of antiviral therapy than are culture or clinical assessment.

Glutathionreduktasemangel mit hämatologischen und neurologischen Störungen

ZusammenfassungIn der vorliegenden Arbeit wird ein neues klinisches Syndrom mit Glutathionreduktasemangel, hämatologischen und neurologischen Störungen beschrieben. 1. Die klinische Manifestation des erblichen Enzymdefektes besteht hämatologisch in einer Thrombocytopenie, nichtsphärocytärer hämolytischen Anämie oder Pancytopenie. Leber und Milz sind stets vergrößert. 2. Der z.T. schweren Pancytopenie ging bei drei untersuchten Kranken ein langfristiger Medikamentengebrauch (Phenobutazonderivate, Primaquine, Resochin) voraus. In schwächerer Form scheint diese jedoch auch schon vorher bestanden zu haben. Nach Absetzen der Präparate kam es bei einem Kranken zu einer vorübergehenden Teilremission des Blutstatus. 3. Neurologisch fanden sich bei allen Enzymdefektträgern Pyramidenbahnzeichen (Babinski-Gruppe) und im Elektroencephalogramm eine leichte bis mäßige Allgemeinveränderung mit vermehrten Theta-Wellen. In einer Familie bestand gleichzeitig eine Oligophrenie. Da der Schwachsinn alle Enzymdefektträger betraf, muß an einen metabolischen Schwachsinn gedacht werden. 4. Biochemische Untersuchungen ergaben, daß es sich nicht um einen Mangel an normaler Glutathionreduktase, sondern um die Bildung einer pathologischen Glutathionreduktase handelt. Das abnormale Enzym zeichnet sich durch eine Verschiebung des pH-Optimums zum Sauren und eine Erhöhung der Halbsättigungskonstanten für GSSG aus. Die Michaeliskonstante für NADPH2 war normal. Von dem Enzymdefekt sind Erythrocyten, Leukocyten und Thrombocyten in gleicher Weise betroffen. Die Glutathionstabilität gegen Acetylphenylhydrazin ist noch normal oder leicht pathologisch, der Heinzkörper-Test stets stark pathologisch. 5. Eine Hemmung des Enzyms durch einen Hemmkörper oder das Fehlen eines stromagebundenen Aktivators konnte unwahrscheinlich gemacht werden. 6. Der Erbgang des Enzymdefektes in einer 15köpfigen Familie war — ebenso wie die Thrombocytopenie und die neurologischen Störungen — autosomal dominant. 7. Therapeutisch brachte die Behandlung mit anabolem Hormon und Nebennierenrindensteroiden keinen sicheren Effekt. Die einmal durchgeführte Milzexstirpation besserte lediglich die Thrombocytopenie, während die nichtsphärocytäre hämolytische Anämie unverändert blieb.Von den drei Kranken mit Pancytopenie starben alle in wenigen Monaten. Alle oben angegebenen Medikamente sollten strikt vermieden werden.SummaryA new syndrome with deficiency of glutathione reductase, hematological and neurological disturbances is described. 1. There will be found thrombocytopenia, nonspherocytic hemolytic anemia or pancytopenia. Liver and splen are always enlarged. 2. Pancytopenia may become dangerous after prolonged therapy with phenopyrine derivates, primaquine, or chloroquine; death may occur. 3. Neurologically all patients with enzyme defect from 4 families showed spastic signs, an increase of-waves in the EEG and in 1 family also oligophrenia (metabolic oligophrenia?). 4. Glutathione reductase deficiency could be shown in erythrocytes, leucocytes and thrombocytes. Also as far as the nervous system is concerned, the enzyme defect seems to be the cause of the disturbances.ThekM-values for GSSG was 2 to 3 times above normal, the pH optimum was shifted from pH 6.8 to 6.4. ThekM for NADPH2 was normal. 5. The GSH content of the erythrocytes was normal also in the patients with the enzyme defect. The GSH stability to acetylphenylhydrazine was normal or slightly pathologic. The Heinzbody test (Beutler test) was always markedly pathologic. 6. There is not much probability for an inhibitor or a deficient, stroma bound activator to be the cause of the decrease of the activity of glutathione reductase. 7. The enzyme defect and the neurologic and hematologic changes was autosomally dominant in the family Bo. (15 members). 8. There is no successful therapy for these patients as yet. Drugs, which can cause the manifestation of the enzyme defect, should be strictly avoided.

Phenylhydrazine-induced lipid peroxidation of red blood cells in vitro and in vivo: monitoring by the production of volatile hydrocarbons.

Human red blood cells and male Sprague-Dawley rats were treated in vitro and in vivo, respectively, with phenylhydrazine in order to determine whether the release of volatile hydrocarbons can serve as a suitable index for phenylhydrazine-induced red blood cell peroxidation. Lipid peroxidation following phenylhydrazine administration (in vitro experiments: dosage calculated at 0.5-50 mM; in vivo experiments: intraperitoneal injection of 2.8 mg/100 g body wt) was monitored by the release of ethane and pentane measured by gas chromatography. Further hydrocarbons such as ethylene, propane, n-butane, iso-butane and iso-butene were monitored to form a basis of comparison. In vitro haemolysis was also determined during the course of incubation. Red blood cell suspensions yielded more than 15-fold concentrations of propane and more than 2-fold concentrations of iso-butane compared to pentane and ethane yields. Haemoglobin solutions also produced propane and iso-butane in the presence of phenylhydrazine, whereas pentane and ethane were not detectable. Time-course studies revealed that ethane and pentane reached maximum in vitro levels after red blood cell suspensions had been incubated for 2 hr whereas the maximum degree of haemolysis (approximately 60%) was attained between 60 and 90 min following the beginning of phenylhydrazine treatment. The dosage did not affect the final degree of haemolysis. Rats treated with phenylhydrazine exhaled greater concentrations of ethane (6-fold increase) and pentane (2-fold increase) compared to control animals. Exhaled propane showed a 30-fold increase in concentration following drug treatment. Our results suggest that the release of pentane and ethane may be useful in assessing red blood cell lipid peroxidation in the presence of phenylhydrazine in vitro and in vivo.

The Relationship Between Lipid Composition of Red Blood Cells and Their Susceptibility to Lipid Peroxidation

Red blood cells from 31 healthy donors were examined for the cholesterol content, the fatty acid composition, and the susceptibility to lipid peroxidation induced by either hydrogen peroxide or phenylhydrazine. Lipid peroxidation was monitored by the release of pentane and ethane. In addition, plasma fatty acids were measured in order to find out, whether plasma and red cell fatty acids were correlated. In experiments with hydrogen peroxide, a significant positive correlation was found between the proportion of arachidonic acid (C 20:4n - 6; r = 0.57, p less than 0.01) and docosahexaenoic acid (C 22:6n - 3; r = +0.71, p less than 0.01), and the release of pentane and ethane, respectively. A significant negative correlation was found between the membrane cholesterol content and the pentane release (r -0.44, p less than 0.05). In experiments performed with phenylhydrazine, red cell membrane lipid composition did not influence the susceptibility of red cells to lipid peroxidation. A close correlation was found between plasma and red cell fatty acids (palmitic acid, r = +0.46, p less than 0.01; linoleic acid, r = +0.41, p less than 0.05; arachidonic acid, r = +0.59, p less than 0.01; docosahexaenoic acid, r = +0.67, p less than 0.01). The results demonstrated that the degree of peroxide-induced oxidation of erythrocyte lipids depends on the content of polyunsaturated fatty acids in the membrane, which on the other hand, is determined by plasma fatty acids. It is suggested that dietary variations may influence the susceptibility of red cells to lipid peroxidation.

Decreased Essential Antioxidants and Increased Lipid Hydroperoxides Following High-Dose Radiochemotherapy

The blood from 19 patients having bone marrow transplantation was examined for the essential antioxidants alpha-tocopherol and beta-carotene as well as lipid hydroperoxides before, at and after bone marrow transplantation (BMT). Conditioning therapy, preceding BMT in order to achieve marrow ablation and immunosuppression, consists of high-dose chemotherapy which is mostly combined with total body irradiation (TBI). In order to see a possible difference between patients with and without additional TBI, we divided the patients up into two groups; patients receiving TBI (RT+) and patients without TBI (RT-). All patients required total parenteral nutrition beginning one week prior to BMT. After conditioning therapy plasma levels of absolute and lipid-standardized alpha-tocopherol and beta-carotene decreased in both groups, presumably as a result of an enhanced breakdown of these antioxidants. The loss of these lipid-soluble antioxidants has to be considered as a possible cause for early post-transplant toxicity. Lipid hydroperoxides increase significantly in the group of patients with additional TBI, whereas the other group, receiving no additional TBI, showed no significant change. We suggest high-dose supplementation of essential antioxidants for patients undergoing BMT.

Hairy Cell Leukaemia: Surface Markers and Functional Capacities of the Leukaemic Cells Analysed in Eight Patients

Summary. In eight patients with hairy cell leukaemia (HCL) peripheral blood cells and in two patients also spleen cells were analysed for surface markers and functional capacities. Only cells containing the tartrate resistant isoenzyme 5 of the acid phosphatase were considered. Hairy cells (HC) of all patients were found to adhere spontaneously to glass and plastic surfaces and to spread after adherence like monocytes. They ingested latex particles of more than 1 μm diameter, but, in contrast to monocytes, did not phagocytose erythrocytes sensitized either by IgM or by IgG antibodies. HC of all patients bore Fc‐receptors with a high binding affinity for aggregated IgG. Using 125I‐labelled F(ab′)2‐fragments of monospecific antibodies in autoradiography, only one light chain type was detected on HC of individual patients. In four patients μ‐ and δ‐chains were simultaneously expressed on HC, whereas in two patients only γ‐chains and in one case only μ‐chains were observed on HC. One patient showed a combination of γ‐ and δ‐chains on his HC. A great variation in density of surface immunoglobulins of HC was observed within individual patients. After removal by capping, surface immunoglobulin reappeared on HC during cell culture, but more slowly than on normal B‐lymphocytes. As shown in two patients by internal labelling, HC secreted immunoglobulin light chains, but no heavy chains. On the basis of these findings the classification of HC as belonging to the B‐cell lineage, rather than to the monocytic lineage, seems to be justified.

Decreased susceptibility of red blood cells to lipid peroxidation in patients with alcoholic liver cirrhosis

Red blood cells from alcoholics with and without liver cirrhosis and control subjects were examined for the susceptibility to lipid peroxidation. Red blood cells of patients with liver cirrhosis were found to be less sensitive to hydrogen peroxide-induced peroxidation measured by a new, reliable and sensitive method: the release of pentane during red blood cell lipid peroxidation. Changes of sensitivity to lipid peroxidation correlated with the severity of the liver malfunction, but not with abnormalities of the lipid composition of red cell membranes which are apparent in patients with liver disease. In alcoholics without liver cirrhosis, only minor changes in the susceptibility of red cells to peroxidation were observed.

Der Hämiglobingehalt in alternden Erythrocyten

ZusammenfassungDer Hämiglobingehalt ist in alternden Erythrocyten höher als in jungen roten Blutzellen. Der Anteil des Hämiglobins am Gesamthämoglobin steigt in einer Erythrocytenpopulation bei intravitaler Alterung von 80 Tagen von 0,01 auf etwa 0,08 an. Der anstieg des Hämiglobins wird von den Verfassern durch den Aktivitätsverlust der DPNH2-liefernden Reaktionen im Erythrocyten, die den Wasserstoff für die Hämiglobinreduktion bilden, erklärt.