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Summary. We studied the influence of human recombinant granulocyte-macrophage colony-stimulating factor (rhGM-CSF) and Candida albicans (CA) components, either alone or in combination, on the proliferation of human bone marrow cells in vitro (colony-forming assay). The number of colonies per 105 bone marrow cells after cultivation with rhGM-CSF [maximal (plateau) colony formation] was: 46.2 ± 9.1 (n= 6); after cultivation with rhGM-CSF in combination with CA components the numbers of colonies were as follows: 0.05 mg proteidml, 33.4 ± 4.6 (n= 3); 0.10 mg protein/ml, 20.8 ± J.6 (n= 3). The mechanisms responsible for this inhibition of colony formation are still unknown. They may be of pathogenetic significance in CA infection. Zusammenfassung. Es wurde der Einflus von Candida albicans (CA)-Bestandteilen auf die Proliferation (Kolonienbildung) menschlicher Knochenmarkzellen unter In-vitro-Bedingungen untersucht (Kolonien-Bildungstest). Die maximale Kolonienbildung pro 105 Knochenmarkzellen nach Kulti-vierung der Zellen allein mit dem rekombinanten Granulozyten-Makrophagen-Kolonien-stimu-lierenden Faktor (rHuGM-CSF) betrug: 46.2 ± 9.1 (n= 6); nach Kultivierung mit dem rHuGM-CSF und zusammen mit CA-Bestandteilen: 0.05 mg Protein/ml Kulturmedium: 33.4 ± 4.6 (n= 3); 0.10 mg/ml: 20.8 ± 3.6 (n= 3). Die Ursache fur diese Hemmwirkung der CA-Bestandteile ist noch unbekannt. Sie konnte bei CA-Infektionen patho-genetisch bedeutsam sein.

Exposure of mammalian cells to UVA in vitro results in fundamental metabolic, functional and structural alterations and after all in necrosis, apoptosis, mutation or restitution. The cascade of biochemical events encloses among other things generation of free radicals, an activation of signal transduction pathways, gene expression and an increased intracellular calcium concentration. Calcium-activated enzymes mediate a part of the processes that cause cell injury after UVA-radiation. Various endogenous and exogenous compounds which interfere with these deleterious chain reactions have a protective effect against UVA-mediated cell damage [1].

We studied the influence of human recombinant granulocyte-macrophage colony-stimulating factor (rhGM-CSF) and Candida albicans (CA) components, either alone or in combination, on the proliferation of human bone marrow cells in vitro (colony-forming assay). The number of colonies per 10(5) bone marrow cells after cultivation with rhGM-CSF [maximal (plateau) colony formation] was: 46.2 +/- 9.1 (n = 6); after cultivation with rhGM-CSF in combination with CA components the numbers of colonies were as follows: 0.05 mg protein/ml, 33.4 +/- 4.6 (n = 3); 0.10 mg protein/ml, 20.8 +/- 3.6 (n = 3). The mechanisms responsible for this inhibition of colony formation are still unknown. They may be of pathogenetic significance in CA infection.

In a double-blind study, 198 patients experiencing recurrent herpes orofacialis were randomly assigned to treatment with either tromantadine hydrochloride (ViruMerz Serol, CAS 53783-83-8) or aciclovir. All patients performed an up to 5-day course of topical treatment beginning on average within 2 h after the first signs of recurrence. The herpes efflorescences and symptoms were assessed daily by the same physician for 14 days, except on weekends, and by the patients each day during the whole observation time. Rapid healing was achieved with both medications. Efficacy was assessed by rating the course of vesicle eruptions, duration until beginning of incrustation and the clinical course of the symptoms (burning, tension, pain). Equivalence between the medication groups was shown by comparative analysis of all evaluation criteria. The global efficacy and tolerability of both medications was rated by the physician as well as by the patients as "good" or "very good" in more than 80% of the cases. The results of this trial show equivalence of both medications in the treatment of recurrent herpes orofacialis, and confirm the good dermal tolerability of the drugs.

Ammonium bituminosulphonate (Ichthyol) inhibits 5-lipoxygenase activity in human polymorphonuclear neutrophils. The inhibition is dose-dependent and occurs at non-cytotoxic concentrations of the drug. This results in a decreased release of Leukotriene B4 from polymorphonuclear neutrophils. Furthermore, when applied to the ear skin of AB/Bln mice pretreated with croton oil, Ichthyol reduces the inflammatory reaction.

Both, ammonium bituminosulfonate (Ichthyol) and Pix lithanthracis reduce the number of ATPase-positive epidermal Langerhans cells (ELC) in the epidermis of BALB/c mice: vaselinum flavum alone vs vaselinum flavum +5% Ichthyol: P less than 0.01; vaselinum flavum vs vaselinum flavum +5% Pix lithanthracis: P less than 0.001. In contrast to this, the number of Ia-positive cells was not changed under identical conditions. These results allow the conclusion that Ichthyol and Pix lianthracis are able to inhibit the enzyme ATPase on the surface of ELC. We infer that inhibition of ELC ATPase may be important in the regulation of ELC function (inhibition of cutaneous contact hypersensitivity).

Water containing high concentrations of magnesium ions (e.g. Dead Sea water) is effective in the treatment of inflammatory skin diseases. Therefore, we examined the influence of Mg2+ on inflammation in allergic contact dermatitis induced by 1-chloro-2,4-dinitrobenzene (DNCB) in BALB/c mice. Animals challenged with 0.125% DNCB in the presence of magnesium chloride (28% and 14%) demonstrated significantly less pronounced contact dermatitis (ear swelling) than did animals challenged with DNCB alone (p less than 0.001 and p less than 0.01). In mice challenged with DNCB in combination with sodium chloride (14%) there was no statistically significant difference in the degree of ear swelling. These results were borne out in 5 patients known to be allergic to nickel, in whom magnesium chloride but not sodium chloride, suppressed nickel sulphate-induced contact dermatitis.

8 patients with confirmed psoriatic arthritis were treated with splenopentin for 12 months. A relief of joint pain could be detected under this treatment both after 6 weeks (improvement of the Ritchie-indices: 44%) and after 12 months (improvement: 28%). In contrast to this, X-ray investigations show a worsening of the disease after 12 months. Therefore we conclude that splenopentin alone is not a sufficient therapeutic drug in patients suffering from psoriatic arthritis.

A PUVA treatment of the recipients of murine tail skin grafts led to a significant prolongation of transplant survival. A possible mechanism of the systemic immunomodulatory effects of ultraviolet light in the organism is the transformation of urocanic acid in the skin from its trans- to the cis-isomer, which acts as a mediator on the immune system. A treatment of graft recipients with cis-, but not trans-urocanic acid also prolonged graft survival. The rejection of secondary grafts and the humoral immune response of mice to sheep erythrocytes were not influenced by cis-urocanic acid. These results implicate that cis-urocanic acid may be get use in clinical organ transplantation and dermatology.

In the last years a new therapy of psoriasis was developed, which consists in a treatment with salt solutions, resembling the water of the Dead Sea, and ultraviolet light (Tomesa-therapy). We studied the influence of the used salt on ATPase positive epidermal Langerhans cells in murine ear skin. An irreversible partial reduction of the Langerhans cell ATPase was found after salt treatment of separated epidermis or of full skin preparations. These results may have implications for the optimization and broader application of this therapy.

Using mononuclear cells (MNC) of patients suffering from psoriasis for production of colony-stimulating factors (CSF) an abnormal colony formation in bone marrow of healthy volunteers was detected (colony-forming assay): Number of colonies per 10(5) bone marrow cells after cultivation with 10(6) MNC of patients suffering from psoriasis vulgaris: 65 +/- 14 (n = 8); psoriasis arthropathica: 63 +/- 11 (n = 8). Control values (10(5) bone marrow cells plus 10(6) MNC of healthy donors): 116 +/- 23 (n = 10). Statistical differences: psoriasis vulgaris: p less than 0.001; psoriasis arthropathica: p less than 0.001. The mechanisms responsible for these abnormalities in hematopoiesis are still unknown. They may be of pathogenic significance in psoriasis.

Perilymphe and Serum von Meerschweinchen wurden mittels Immun- and Polyakrylamidgel-Elektrophorese untersucht. Die Molekulargewichte der Perilymphproteine wurden durch Elektrophorese in einem linearen Polyakrylamidgelgradienten (3–20%) ermittelt. Die Immun- and Polyakrylamidgel-Elektropherogramme von aquivalenten Mengen Perilymph- and Serumprotein sind weitgehend ahnlich. Der immunologische Nachweis der Perilymphproteine weist darauf hin, daβ es sich urn Serumproteine handelt. Fur eine eindeutige Identifizierung der einzelnen Proteine fehien monospezifische Antiseren gegen die einzelnen Proteine des Meerschweinchens. Die Molekulargewichtsermittlung zeigt, daβ in der Perilymphe auch relativ hochmolekulare Proteine (Molekulargewichtsabschatzung im Bereich von 67000-290000) nachweisbar sind.

Yeast PFK had a sedimentation coefficient of 16.7 S both in the absence and in the presence of ATP, and did not dissociate even at very low protein concentrations. Sodium dodecyl-sulphate caused dissociation of the protein to sub-units of 3.2 S. The effects of pH on substrate affinities are described. In the presence of UTP, acting as non-inhibiting phosphate donor, the behaviour of the enzyme towards F-6-P was co-operative, with a Hill coefficient of 2.2.

Yeast phosphofructokinase binds to Blue Dextran 2000® with remarkable high affinity. Whereas phosphofructokinase from muscle and erythrocytes also have high affinity to Blue Dextran, aldolase, hexokinase, pyruvate kinase from muscle and glyceraldehyde-3-phosphate dehydrogenase show no affinity to this chromophore. The chromophoric component of Blue Dextran seems to be responsible for these interactions, because chromophore-free dextran 2000 does not show any affinity to phosphofructokinases. Blue Dextran immobilized by fixation in cross-linked polyacrylamide gel can be used as a valuable tool for affinity chromatography of phosphofructokinase. The conditions of binding, namely pH dependence, effects of ionic strength and of enzyme effectors have been studied using gel-fixed phosphofructokinase. ATP (2–3 mM) but not ITP (up to 5 mM) specifically splits the enzyme chromophore complex. With non-inhibitory concentrations of ATP Blue Dextran inhibits the enzyme very strongly showing competition with respect to ATP. No correlation between inhibition by Blue Dextran and fructose-6-phosphate concentration could be detected. A model is proposed for the interactions of Blue Dextran with phosphofructokinase, postulating binding of the chromophore to the ATP binding sites of the enzyme.

According to Id according to Liebe et al. [2] the enzyme may contain subunits of 60,000 Daltons. While these studies were in progress Wilgus et al. [4] presented evidence for the existence of a polypeptide chain molecular weight of 1 .O X 10’ and considered the products of splitting just described as proteolytically degraded fragments. In this contribution the values of physical molecular parameters of yeast phosphofructokinase are presented since they are necessary for a precise calculation of its molecular weight. Furthermore, the subunit structure of yeast phosphofructokinase is reexamined in more detail. Finally, evidence is presented that the true molecular weight of undegraded yeast phosphofructokinase is apparently higher than hitherto assumed.

A new method for molecular weight determination using polyacrylamidegel electrophoresis in a linear gel-concentration-gradient (3–20°) is described. Plotting the log of molecular weights of several standard proteins against distance of migration or against 3 log of rate of n-dgration reveal linear relationships in ranges of 50.000 – 200.000 or 100.000 – 400.000 Daltons respectively. On this basis, a simple method for molecular weight determination of proteins (accuracy ± 5°) has been devised. The method can also be applied for an individual protein in protein mixtures using specific staining procedures.

Phosphofructokinases from mammalian tissues occur in different association states under a variety of experimental conditions [l-6]. These states depend on protein concentration and were found to be influenced by substrates and effecters as well as by changing pH values and ionic strengths. Layzer and coworkers [7,8] described some properties of partially purified erythrocyte phosphofructokinase including a concentration-dependent sedimentation behaviour of the enzyme using density gradient centrifugation. The present paper reports the existence of a series of oligomeric forms of human erythrocyte phosphofructokinase in highly purified enzyme preparations. These are correlated with a product of splitting of the enzyme having a molecular weight of 104,000 which could be characterized by sodium dodecylsulphate (SDS) polyacrylamide gel electrophoresis.

Yeast phosphofructokinase exists in several enzymically active, interconvertible forms with molecular weights of 180,000, 370,000, 570,000 and 750,000. With disc-electrophoresis catalytically active aggregation products with molecular weights of more than one million can be detected. In alkaline media fragmentation of phosphofructokinase occurs, leading to a variety of catalytically inactive products. These seem to be oligomers of subunits with 60,000 daltons. A model of the complex subunit structure of yeast phosphofructokinase is suggested. The various catalytically active forms of the enzyme are considered as polymers of 180,000 monomers, which themselves are enzymatically active and which are composed of three 60,000 subunits.

SUMMARY Interferon-like activity could be detected in the sera of nineteen psoriatic patients (interferon titres between 8 and 64 i. u./ml), whereas healthy controls were negative. Following PUVA treatment the interferon levels increased significantly (interferon titres between 32 and 128 i. u./ml). An enhanced interferon production could explain some of the immunological phenomena connected with PUVA treatment.

The results reported in connection with previously published data suggest the importance of increased lipid peroxidation in the epidermis after Dithranol treatment. Malonaldehyde, one of the main products of lipid peroxidation, reacts with epidermal phosphofructokinase and glucose-6-phosphate dehydrogenase to cause enzyme inactivation by cross-linking of the enzyme protein demonstrated by dodecyl sulphate electrophoresis. Enzyme inactivation leads to an inhibition of hexose monophosphate shunt and glycolysis. The effect on energy donating metabolism in psoriatic lesions is discussed.

The wound-healing agent Dalargin, a hexapeptide and varios of its cleavage products were tested in a wound-healing model on the basis of granulation in rat. Three days after wounding (p.op.) an increase in granulation was observed at Dalargin concentrations of 10 micrograms/ml and 50 micrograms/ml. Seven days p.op. an increase was observed already at a Dalargin concentration of 5 micrograms/ml. The cleavage products of Dalargin had no stimulating effect on granulation. A histologic differentiation of the different cell parts in the granulation tissue led to an increased total number of cells. The concentration of 5 micrograms/ml induced both an increase in the number of endothelial cells and fibroblasts, the concentration of 50 micrograms/ml induced just an increase of the number of endothelial cells. The possible bases of the mode of operation are being discussed. The stimulating effect on wound healing as described could be confirmed by the results obtained in our animal experiments.

Zur Reinigung von Roggenpollenextrakten (Secale cereale) wurden saulenchromatographische Untersuchungen durchgefuhrt. Durch stufenweise Erhohung der Ionenstarke lassen sich an DEAE-Zellulose aus dem nicht dialysierbaren Ruckstand des wasrigen Extraktes mehrere Fraktionen gewinnen (Abb. 1). Immunelektrophorese, Immunodiffusion nach Ouchterlony und Polyakrylamid-Gelelektrophorese zeigen jedoch, das diese Fraktionen Antigen- bzw. Proteingemische unterschiedlicher Heterogenitat sind (Abb. 2 und 3, Tabelle 1). Sie lassen sich auch durch Rechromatographie an Sephadex G-100 nicht in homogene Fraktionen trennen. Die meisten Fraktionen zeigen im direkten Intrakutantest an nicht behandelten Roggenpollenallergikern Allergenaktivitat (Tabelle 1). Die Auswahl bestimmter Fraktionen fur eine weitere Trennung ist durch die quantitative Ungenauigkeit des Intrakutantestes erschwert. NDS-gelelektrophoretische Untersuchungen unter Verwendung des Proteinaseinhibitors Phenylmethylsulfonylfluorid weisen auserdem darauf hin (Abb. 4), das in den Roggenpollenextrakten Proteinasen enthalten sind, die sich wahrscheinlich nur schwer von den ubrigen Pollenproteinen trennen lassen und das Proteinmuster verandern. Im Intrakutantest ist jedoch auch nach einjahriger Aufbewahrung der Extrakte keine Veranderung ihrer Allergenpotenz nachweisbar.

Rabbits were immunized using isolated and dissolved specific immune complexes obtained from patients suffering from early syphilis. After immunization the rabbits were infected intradermally with living Treponema pallidum. Some of them did not develop lesions at the infection sites. In comparison to controls there was a delayed and reduced production of antibodies. The effect of immunization may be induced by a treponemal protein antigen as a constituent of the specific immune complexes.

Immune complexes could be measured in sera with the use of plastfixed Clq and 125J-labeled anti-Ig-antibody. The sensitivity of the system is 0.1 microgram aggregated human IgG, the working range is between 0.1 and 10 micrograms per 0.5 ml. In 58% of the sera from patients with systemic lupus erythematosus was found an increase in the immune complex content.

By means of a polyethylene glycol precipitate method [6] circulating immune complexes were examinated in increased concentration in 56 percent of the sera from psoriatic patients. The effect of photochemotherapy with external administration of 8-methoxypsoralen followed by irradiation with UVA was investigated. In 12 of 13 cases the serum immune complex concentration after the therapy was normal.

Perilymph and serum of guinea pigs were investigated using immunoelectrophoresis and polyacrylamide gel electrophoresis. The molecular weights of the perilymph proteins were estimated by electrophoresis in a linear polyacrylamide gel gradient (3-20%). The immunoelectropherograms and the polyacrylamide gel electropherograms of equivalent amounts of perilymph protein and serum protein are nearly the same. The immunological detection of the perilymph proteins indicates that they are serum proteins. The clear identification of the individual proteins fails because of missing monospecific antisera against the individual proteins of guinea pig. High-molecular proteins (range of the molecular weight estimation 67 000-290 000) are also detectable in the perilymph according to the molecular weight estimation.

The maturation effect of DA-SP5 on a spleen cell population depleted of immunocompetent, histami nereceptor-bearing lymphocytes was examined. Conjugates of histamine and the A-chain of the mistletoe lectin I killed in vitro these cells and abolished, as shown in a previous paper, their humoral and cellular capacity by around 90%. Sublethally irradiated mice were reconstituted with the remaining cells and treated with DA-SP5 (3 intraperitoneally injections of 10 micrograms DA-SP5 per week). The strong accelerating action of DA-SP5 on the restoration of the antibody response is interpreted as an effect of DA-SP5 on the maturation of B- and T-cells under in vivo conditions.

Sublethally irradiated AB/Bln mice (500 cGy) were treated with splenotritin (fragment 32-34 of the splenic hormone splenin) and tested for their capability of producing antibodies against target sheep red blood cells. In x-ray plus splenotritin treated mice antibody forming cells were found earlier and in a higher frequency than in mice treated with x-rays alone.

Epidermal Langerhans cells (ELC) are bone marrow-derived immune cells that are important in allergic contact dermatitis. We examined the influence of calcium transport inhibitors, lanthanum and diltiazem hydrochloride, on allergic contact dermatitis induced by 1-chloro-2,4-dinitrobenzene (DNCB) in BALB/c mice. Systemic lanthanum at a dose of 0.08 mg/kg and topical lanthanum (50 microliters of 10% solution) were given 5 d before DNCB sensitization. Systemic diltiazem (30 mg/kg per dy) was given for 3 d during sensitization with DNCB. In all animals, challenge with topical DNCB to the ear skin was performed 5 d after sensitization and ear swelling was measured. Twenty four hours post-DNCB challenge, animals receiving systemic lanthanum demonstrated a 56% decrease in contact hypersensitivity (ear swelling) compared with non-lanthanum-treated animals (0.08 +/- 0.03 mm vs 0.18 mm +/- 0.02 mm, p less than 0.01). Topical lanthanum produced a 58% decrease in contact hypersensitivity (0.20 +/- 0.02 mm vs 0.41 +/- 0.03 mm, p less than 0.01). The DNCB-induced ear swelling also resolved more quickly in animals treated with lanthanum. Systemic diltiazem produced a 67% decrease in ear swelling (0.05 +/- 0.01 mm vs 0.15 +/- 0.02 mm, p less than 0.001). A decrease in epidermal Langerhans cell density of 13 to 14% was produced by systemic lanthanum, detected by both ATPase staining and Ia staining, respectively (p less than 0.05). Approximately 20% of the Langerhans cells were morphologically abnormal, having become "rounded," and lacking normal dendritic processes. From these results, we infer that calcium transport across the cell membrane of ELC may be important in the regulation of their function. Lanthanides and other calcium-channel blockers may be useful pharmacologic agents to probe these phenomena.