Reiner Scherer

UVA‐induced inhibition of proliferation of PHA‐stimulated lymphocytes from humans treated with 8‐methoxypsoraien

Lymphocytes from healthy human donors were used as a model system for studying the combined effect of 8-methoxypsoralen (8-MOP) plus UVA. Two hours after oral administration of therapeutic doses of the drug enough 8-MOP was taken up in vivo by the circulating peripheral lymphocytes to cause significant inhibition of phytohaemagglutinin induced lymphocyte proliferation when the cells were exposed in vitro to UVA irradiation. The inhibition of proliferation as monitored by a reduced 3H-thymidine incorporation into cellular DNA was shown to be UVA-dose dependent. Control cultures of 8-MOP containing lymphocytes which were not UVA irradiated showed normal blast transformation. In lymphocytes obtained from the same donors prior to 8-MOP intake, PHA-induced lymphocyte transformation was not impaired by UVA irradiation. In 8-MOP containing lymphocytes exposed repeatedly to UVA during the 72 h culture period a cumulative effect of irradiation could be observed. A varying sensitivity towards 8-MOP plus UVA was noted when the cells were irradiated at different times after PHA stimulation. The cells were most vulnerable to UVA irradiation during the DNA-synthesis phase of the proliferating lymphocytes. The results suggest that dermal inflammatory infiltrates containing locally proliferating lymphocytes are influenced by systemic photochemotherapy since UVA penetrates well into the dermis.

Immunoelectronmicroscopic examination of early lesions in histamine induced immune complex vasculitis in man

In 4 cases of allergic vasculitis circulating immune complexes (IC) were demonstrated. Spontaneous and histamine induced vascular changes were studied by immunofluorescence microscopy. The early events in IC vasculitis were investigated at the ultrastructural level by immunoelectronmicroscopy using the peroxidase‐antiperoxidase multistep technique.

The human peripheral lymphocyte—a model system for studying the combined effect of psoralen plus black light

ZusammenfassungEs wurde die Wirkung von Psoralen zusammen mit langwelligem ultraviolettem Licht (UVA) auf den3H-Thymidin-Einbau PHA-stimulierter humaner Lymphocyten untersucht. Die PHA-induzierte Lymphocytentransformation wurde durch Psoralen plus UVA gehemmt, abhängig sowohl von der Psoralenkonzentration als auch der Einwirkdauer der UVA-Bestrahlung. Die Hemmung der DNA-Synthese in den stimulierten Lymphocyten erfolgte bei Psoralenkonzentrationen wie sie auch im Serum von Patienten erwartet werden können, die sich einer systemischen Photochemotherapie unterziehen. Ohne UVA-Bestrahlung war bei diesen Psoralenkonzentrationen kein Effekt auf die DNA-Synthese nachweisbar. Ebenso erwies sich alleinige UVA-Bestrahlung ohne Psoralenzusatz als wirkungslos auf das Wachstum PHA-stimulierter Lymphocytenkulturen.SummaryThe effect of psoralen plus long wavelength ultraviolet light (UVA) on3H-thymidine uptake of PHA stimulated human lymphocytes was investigated. PHA induced lymphocyte transformation was inhibited by the combined action of psoralen and UVA irradiation in a dose related manner. Inhibition of DNA-synthesis occurred at concentrations of psoralen that can be expected in the serum of patients treated by systemic photochemotherapy. No effect was noted at these psoralen concentrations in the absence of UVA irradiation. Also did UVA irradiation in the absence of psoralen not inhibit3H-thymidine incorporation into PHA stimulated lymphocytes.

Immunoelectronmicroscopical demonstration of in vivo bound complement C 3 in psoriatic lesions

SummaryThe presence of in vivo bound complement (C3) within the stratum corneum of 4 psoriatic cases is demonstrated ultrastructurally by the use of the peroxidase—antiperoxidase multistep technique. The positive reaction product was located in the intercellular space and on the surface of the parakeratotic cells within the horny layer. The membranes of the horny cells were not altered and peroxidase granules could also not be detected within their cytoplasm. The explanation of this finding as well as its pathogenetic significance is discussed.ZusammenfassungBei 4 Patienten mit Psoriasis vulgaris wurde in vivo gebundenes Komplement (C3) innerhalb des Str. corneum von Psoriasisherden durch Anwendung der Peroxidase—Antiperoxidase Mehrstufentechnik geortet. Reaktionsprodukte fanden sich sowohl intercellulär als auch den Zellmembranen der parakeratotischen Zellen der Hornschicht angelagert. Diese Befunde werden hinsichtlich ihrer pathogenetischen Bedeutung diskutiert.

Erythema elevatum diutinum@@@Erythema elevatum diutinum: II. Immunelektronenmikroskopische Untersuchung der leukocytoklastischen Vaskulitis in einer Intrakutanreaktion mit Streptokokkenantigen@@@II. Immunoelectronmicroscopical study of leukocytoclastic vasculitis within the intracutaneous test reaction induced by streptococcal antigen

SummaryA leukocytoclastic vasculitis was induced by intracutaneous injection of streptococcal antigen in a patient with erythema elevatum diutinum (E.e.d.). The immunoelectronmicroscopical demonstration of C3 was performed by use of the peroxidase-antiperoxidase multistep technique 24 h after the injection of the antigen.Deposits of C3 were found between endothelial cells, on the outer surface of endothelial cells, pericytes, and smooth muscle cells, as well as within the multilayered basal lamina of small vessels. Intact and disintegrating neutrophils accumulate within the vessel walls and in their surroundings. Necrosis and fibrin deposition are present in advanced stages.The findings demonstrate the sequence of events in leukocytoclastic vasculitis at the ultrastructural level. They also support the hypothesis that in E.e.d. an Arthus type reaction induced by bacterial antigens may be of pathogenetic significance.ZusammenfassungEine leukocytoklastische Vaskulitis wurde durch intrakutane Injektion von Streptokokkenantigen bei einer Patientin mit Erythema elevatum diutinum (E.e.d.) ausgelöst. Der immunelektronenmikroskopische Nachweis von C3 in dieser Reaktion wurde 24 h nach der Injektion der Antigens mit Hilfe der Peroxydase-Antiperoxydase-Mehrstufentechnik durchgeführt.C3-Niederschläge fanden sich in der Intercellularfuge zwischen Endothelzellen, Pericyten und glatten Muskelzellen und der aufliegenden Basallamina sowie zwischen den Duplikaturen der mehrschichtigen Basallamina kleiner Gefäße. Intakte und zerfallende Neutrophile durchsetzen die Gefäßwände und die Gefäßumgebung. Es resultieren Nekrose und Fibrinablagerung.Die Befunde zeigen die Sequenz der Ereignisse bei leukocytoklastischer Vaskulitis auf dem ultrastrukturellen Niveau und stützen gleichzeitig die Hypothese, daß eine durch Bakterienantigen ausgelöste Reaktion vom Arthus-Typ bei E.e.d. pathogenetisch bedeutsam ist.

[The specific effect of plasma proteins in erythrocyte sedimentation. An analysis of the correlation coefficients between ESR and the concentration of twenty individual plasma proteins in healthy and sick persons, with special reference to myocardial infarction (author's transl)].

Summary1. Spearmans rank correlation coefficient between erythrocyte sedimentation rate and the concentration of twenty individual plasma proteins was determined in 72 persons including 37 patients with recent myocardial infarction. The highest correlation coefficients with ESR could be demonstrated in the following proteins: fibrinogen, α-1-acid-glycoprotein, α-2-macroglobulin, α-1-antitrypsin, coeruloplasmin, Ig M. The closest correlation with ESR was found, when the molar concentrations of fibrinogen, α-2-macroglobulin and Ig M were summed up.2. Subdivision of the examined group of patients according to their diagnosis showed that the degree of correlation with ESR in many plasma proteins essentially depends on the composition of the studied group of patients. The patients with myocardial infarction, with pneumonia and those with neoplasma all showed distinctly different patterns of correlation. These obviously reflect underlying changes in the concentrations of the studied proteins, that seem to be specific to the respective disease.3. In 4 patients with myocardial infarction the changes in plasma protein concentrations together with ESR were followed over a period of 26 days after the infarction.Zusammenfassung1. Bei 72 Probanden, davon 37 Herzinfarktpatienten wurde der Spearmansche Rangkorrelationskoeffizient von BKS-Geschwindigkeit und den Konzentrationen von zwanzig einzelnen Plasmaproteinen bestimmt. Die höchsten Korrelationskoeffizienten wurden in der Reihenfolge ihrer Nennung gefunden für: Fibrinogen, Orosomukoid, α-2-Makroglobulin, α-1-Antitrypsin, Coeruloplasmin, Ig M. Die Summe der molaren Konzentrationen von Fibrinogen, α-2-Makroglobulin und Ig M wies gegenüber der BKS-Geschwindigkeit die engste Korrelation auf.2. Die Aufschlüsselung des untersuchten Kollektivs nach der Diagnose ergab, daß für viele Plasmaproteine der Grad der Korrelation zur BKS-Geschwindigkeit wesentlich von der Zusammensetzung des untersuchten Patientengutes abhängt. Die Gruppe der Patienten mit Herzinfarkt, mit Pneumonie und mit Neoplasma wiesen jeweils deutlich verschiedene Korrelationsspektren auf, denen offenbar krankheitsspezifische Änderungen der Konzentrationen der Plasmaproteine zugrunde liegen.3. Bei 4 Herzinfarktpatienten wurden über einen Zeitraum von 26 Tagen die Änderungen der Plasmaproteinkonzentrationen im Zusammenhang mit der BKS-Geschwindigkeit verfolgt.

Partially hepatectomized rats : a model for the study of the effect of toxins on the plasma protein profiles of nascent hepatocytes

A useful framework is proposed for unifying the synthesis of plasma proteins and their degradation by, or release from, liver cells of intact and partially hepatectomized rats, in which synthesis and release of acute-phase plasma proteins occur in synchrony with the internalization and catabolism of plasma and extracellular proteins. The catabolism of proteins and other hepato-intracellular glycoproteins during sepsis or trauma is essential to provide constituent amino acids and carbohydrates for the synthesis of acute-phase plasma proteins. Increases in the plasma levels of acute-phase response proteins in sham-operated rats reached a maximum between 1 and 2 d after mock surgery, and had returned virtually to control levels within 6 d. By contrast, acute-phase proteins in the plasma of partially hepatectomized rats were decreased by 10-20% of their initial values after 24 h. A maximum acute-phase response on d 7 after the operation was characterized by an increase of 181, 445, and 19% for alpha-1-acid glycoprotein, hepatoglobin, and hemopexin, whereas other acute-phase proteins remained below control levels, for example, by 11, 25, and 38% for albumin, transferrin, and prealbumin, respectively. This delayed response suggests that the nascent liver cells had inherited the capacity of the parent cells to respond to inflammatory signal and had synthesized acute-phase plasma proteins. Accordingly, a time frame for the application of toxin to nascent hepatocytes is suggested. An increased activity (300 +/- 50%) for both bound and free neuraminidase in remnant liver tissue 19 h post partial hepatectomy suggested that hepatic regenerating factor(s) were produced in liver tissue via the hepatic bound and/or free neuraminidase-mediated desialylation of humoral substrates. By contrast, circulating levels of lysosomal enzymes alpha-fucosidase and beta-N-acetyl-D-glucosaminidase were increased marginally after 24 h but had returned nearly to control levels after 7 d, suggesting that lysosomal acid hydrolases do not play a major role in regenerative DNA synthesis, mitosis, or in the synthesis of acute-phase plasma proteins.

Lecithinabbau und Cholesterinveresterung im Humanserum

SummaryThe physiological role of lecithin-cholesterol acyltransferase as is known so far is the transport of cholesterol from peripheral tissues including blood cell membranes to the liver. Another function seems to consist in the stabilization of the lipoprotein structure. This is suggested by clinical findings in patients with either genetic or secondary LCAT-deficiency. LCAT-activity depends on the functional state of the liver which is responsible for the enzyme synthesis. The esterifying activity is also greatly influenced by the quantity and the lipid composition of lipoprotein substrate. Therefore, hyperlipidaemia is often associated with increased LCAT-activity.ZusammenfassungDie physiologische Rolle der Lecithin-Cholesterin Acyltransferase (LCAT) scheint im Transport von Cholesterin von den peripheren Geweben einschließlich der Membranen der Blutzellen zur Leber zu liegen. Eine weitere Funktion ist vermutlich die Stabilisierung der Lipoproteinstruktur. Dies geht aus klinischen Befunden bei Patienten hervor, die an einem primären genetischen oder sekundärem LCAT-Mangel leiden. Die LCAT-Aktivität hängt vom funktionellen Zustand der Leber ab, die für die Synthese des Enzyms verantwortlich ist. Die Veresterungsaktivität wird zusätzlich stark von der Menge und der Lipidzusammensetzung des Lipoproteinsubstrates beeinflußt. Aus diesem Grund ist die LCAT-Aktivität bei Hyperlipidämien oft gesteigert.

[Erythema elevatum diutinum. I. Electron microscopy of a case with extracellular cholesterosis (author's transl)].

First described by Hutchinson in 1878, erythema elevatum diutinum (EED) is a rare disease characterized by persistent red, raised nodules and plaques in a symmetrical, acral distribution. The disease occurs equally in the sexes, primarily in the 30- to 60-year age group. We report a representative case of EED which responded to current agents, and briefly review the status of the disease.

Der Mechanismus der Blutkörperchensenkung. XVII Über den Zusammenhang und die diagnostische Bedeutung der Lecithin-Cholesterin-Acyltransferase-Aktivität im Humanserum und der Wärmeinaktivierung der Blutkörperchensenkung

SummaryThe activity of the lecithin-cholesterol-acyltransferase (LCAT) was determined in 80 healthy men and women, 13 pregnant women, and various patients mostly with neoplastic or hepatic diseases. Lipoproteins with32P-3H-labelled sojabean-lecithin were used as substrate. Significant influences of age, sex and diseased states could be observed:The enzyme activity was more elevated in young men up to 30 years than in women of the same age, it was also higher in women over 40 years compared to that in younger women. The LCAT activity and the lysolecithinase activity were increased in pregnant women. The LCAT activity was decreased in diseases affecting the protein synthesis capacity of the liver like cirrhosis or advanced cancer.The inhibition of accelerated erythrocyte sedimentation after incubation of plasma at 37° is caused by an increase in plasma lysolecithin concentration due to LCAT activity.The clinical importance of differences in the inhibition of accelerated erythrocyte sedimentation in neoplastic and inflammatory diseases is discussed. Contradictory findings in earlier investigations on this subject can be explained on the basis of the differences observed in LCAT activity due to age and sex, variations in the lysolecithinase-activity, and plasma albumin concentration.ZusammenfassungDie Enzymaktivität der Lecithin-Cholesterin-Acyltransferase (LCAT) wurde bei 80 gesunden Männern und Frauen, 13 Schwangeren und bei verschiedenen Patienten mit neoplastischen oder hepatischen Erkrankungen untersucht. Bei dem Test wurden Lipoproteine mit32P-3H-markiertem Sojabohnenlecithin als Substrat verwendet. Es konnten signifikante Einflüsse von Alter, Geschlecht und Krankheit auf die Enzymaktivität beobachtet werden:Die LCAT-Aktivität war bei Männern bis 30 Janre höher als bei Frauen der gleichen Altersstufe, ebenso war sie bei Frauen über 40 Jahre höher als bei jüngeren Frauen. Bei den Schwangeren waren sowohl die LCAT-Aktivität als auch die Lysolecithinase-Aktivität gesteigert. Krankheiten, die die Proteinsynthesekapazität der Leber herabsetzen wie Cirrhose oder fortgeschrittene neoplastische Erkrankungen, gingen mit einer erniedrigten LCAT-Aktivität einher.Die Hemmung einer beschleunigten Blutkörperchensenkung nach Inkubation des Plasmas bei 37° wird durch die Zunahme der Plasmalysolecithinkonzentration bewirkt, die ein Ergebnis der LCAT-Aktivität ist. Die klinische Bedeutung von Unterschieden in der Hemmbarkeit von beschleunigten Blutkörperchensenkungen bei neoplastischen und entzündlichen Erkrankungen wird erörtert. Widersprüchliche Befunde in früheren Untersuchungen zu diesem Thema können durch die beobachteten geschlechts- und altersabhängigen Unterschiede in der LCAT-Aktivität, sowie durch die Einflüsse von Lysolecithinase-Aktivität und Plasmaalbuminkonzentration auf die Senkungshemmung erklärt werden.