Immunomodulating efficacy of combined administration of galactoside-specific lectin standardized mistletoe extract and sodium selenite in BALB/c-mice.

The immunomodulating abilities of commercially available mistletoe extract standardized for the galactoside-specific lectin (mistletoe lectin-1, ML-1) and sodium selenite (Se) were evaluated in BALB/c-mice and compared to non-treated control animals. Following the optimal schedule of administration (ML-1: 1ng/kg BW; days 1, 2, 3, 5 and Se: 3,5 micrograms/kg BW for 7 subsequent days before evaluation) yielded enhanced counts (thymocytes; peritoneal macrophages, MO; peripheral blood leukocytes; lymphocytes, PBL; monocytes, PBM) and activities (CD-25 positive PBL, MAC-3 positive PBM) of desired immune cells reaching statistical significance for most parameters. However, combined administration of ML-1 and Se proved to be superior to monotherapy since immune cell counts (thymocytes, leukocytes, PBL, PBM) and activities (CD-25 positive PBL) exceeded values obtained after monotherapy. These data are in favour of therapeutical strategies in complementary oncology and suggest that the combination of defined immunomodulating substances might positively enhance the efficacy.

Immunoprotective activity of the galactoside-specific lectin from mistletoe after tumor destructive therapy in glioma patients.

35 patients suffering from malignant stage III/IV glioma were enrolled into a prospectively randomized clinical trial. All patients were provided with standard oncologic treatment (neurosurgery, radiation, basic clinical care according to protocol and indication) and randomly divided into a) treatment group: receiving subcutaneous injections of a ML (a galactoside-specific lectin from mistletoe) standardized mistletoe extract, 1 ng ML-1/kg BW, twice a week for 3 months, starting on day 1 post surgery, b) control group: without additional complementary treatment. Immunophenotyping of peripheral blood leukocytes was done by flow cytometry (pre surgery; day 1, week 1, month 3 and 6 post surgery) to evaluate the immunomodulating capacity of ML-1 standardized mistletoe extract. Standard tumor destructive treatment of glioma proved to be suppressive for peripheral blood lymphocytes, since all subsets tested revealed statistically significant down regulation. Unlike the lymphocyte counts and activities of patients from the control group who gave preoperative values after 3-6 months), mistletoe treatment induced a statistically significant up regulation of cell counts (CD-3, CD4, CD-8 cells) and activities (CD-25, HLA/DR positive cells) after 3 months, as compared to preoperative values. Obviously, a strong immunoprotective/immunostimulatory effect was induced by the treatment of glioma patients with ML-1 standardized mistletoe extract which correlated with an improved quality of life, as determined by a standard questionnaire (Spitzer).

Liver lectin blocking with D-galactose to prevent hepatic metastases in colorectal carcinoma patients.

Animal experiments in BALB/c-mice and in DBA/2-mice confirmed that lectin blockade with D-galactose containing receptor analogues can inhibit metastatic spread into the liver. The number of liver colonies of inoculated tumor cells was significantly reduced after D-galactose treatment as compared to animals of control group. Based on experimental investigations 193 colorectal carcinoma patients (UICC stages I-III) were enrolled in a prospectively randomized clinical trial. 93 patients were treated perioperatively with D-galactose- (treatment group: 1.5 g/kg body weight and per day) or D-glucose containing electrolyte infusions (control group: n = 100). Significant side effects were not observed. There were no cases of perioperative mortality. The overall complication rate was 7.3%. Since tumor stages were unequally distributed, analysis was performed in strata. Patients were observed for a total of 6237 months. Differences in overall survival and survival free of recurrence and hepatic metastases were negligible for stages I and II. For stage III carcinoma patients (n = 75) analysis of survival free of hepatic metastases revealed a shift to delayed events (i.e. hepatic metastases or death) after D-galactose treatment within 24 months following surgery. In patients with stage III carcinoma there was an indication for an overall benefit in survival after D-galactose treatment (p = 0.102).

Immunostimulation by propionibacteria--effects on immune status and antineoplastic treatment.

Experimental studies were performed to investigate further the effects of immunotherapy with Propionibacterium avidum KP-40 on thymocyte proliferation, maturation and emigration in BALB/c-mice. Thymus weight and thymocyte counts, especially cells presenting the immature or cytotoxic/suppressor phenotype were significantly increased. Due to enhanced emigration, peripheral blood lymphocyte and monocyte counts as well as expression of activation markers were significantly upregulated. The antimetastatic effect of Propionibacterium avidum KP-40 was demonstrated in BALB/c-mice, where RAW 117-H10 lymphosarcoma liver colonization was significantly reduced after immunostimulation. Clinical investigations proved that surgical treatment of colorectal carcinoma induced an evident decrease of peripheral blood lymphocytes as compared with preoperative counts. However, single preoperative Propionibacterium avidum KP-40 administration induced a considerable increase of peripheral white blood cell counts, especially lymphocytes. Clinical effects of preoperative immunostimulation by Propionibacterium granulosum KP-45 were investigated in a prospective randomized trial in colorectal carcinoma patients. Positive effects on survival time, local tumor recurrence and distant metastasis could be demonstrated in stages I and II, whereas no advantage of immunotherapy was found in advanced stages III and IV. A recent prospective randomized clinical trial was initiated on the quality of life of colorectal carcinoma patients. Three months after surgical treatment negative effects could not be determined after immunotherapy. Quality of life even proved to be better in patients with abdominoperineal resection as compared to non Propionibacterium avidum KP-40 treated control patients.

Effects of the beta-adrenergic agonist clenbuterol in cows: lipid metabolism, milk production, pharmacokinetics, and residues.

The effects of the beta-adrenergic agonist clenbuterol on lipid metabolism, milk production, pharmacokinetics, and residues were studied in six lactating Brown Swiss cows. Four of these were treated with the growth-promoting dose of 5 micrograms of clenbuterol/kg of BW, mixed within the concentrate, and administered twice a day for 3 wk. The remaining two cows served as controls. All animals were in their third phase of lactation and were fed diets containing corn silage, hay, and concentrate according to individual milk production level and body weight. Milk and blood samples were collected for analysis following a rigid time schedule. Milk production and milk contents (triglycerides, protein, and lactose) were quantitatively identical in both treated and control animals, whereas significant qualitative changes occurred in the fatty acid composition of milk lipids in clenbuterol-treated animals. Compared with the controls, the relative amount (percentage of total fatty acids) of unsaturated fatty acids, particularly oleic acid (cis-delta 9-octadecaonate, 18:1), increased considerably with a simultaneous decrease of the transition chain length fatty acids (lauric acid [12:0] and myristic acid [14:0]). Plasma glucose and FFA concentrations were elevated. Concentrations of insulin-like growth factor I remained unchanged. We conclude that the physiological effect of clenbuterol is limited to the repartitioning effect in body composition, and that milk production is affected only slightly by clenbuterol. An immediate increase of clenbuterol concentrations in plasma (3.4 +/- 2.0 ng/mL) and milk (10.8 +/- 4.7 ng/mL) could be observed at the commencement of treatment. Clenbuterol concentration peaked after 10 d and remained constant until the end of the treatment.(ABSTRACT TRUNCATED AT 250 WORDS)

A plant lipid and the platelet-activating factor stimulate ATP-dependent H(+) transport in isolated plant membrane vesicles.

A plant lipid was isolated from zucchini (Cucurbita pepo L.) membranes and from soybean (Glycine max [L.] Merr) phospholipids by thinlayer chromatography and further purified by high-performance liquid chromatography. This plant lipid was chromatographically very similar to the platelet-activating factor, an ether phospho-lipid with hormone-like properties found in mammals. Both the plant lipid and the platelet-activating factor stimulated ATP-dependent H(+) transport in isolated membrane vesicles from zucchini hypocotyls.

Effect of immunomodulation with galactoside-specific mistletoe lectin on experimental listeriosis.

BALB/c-mice (n = 10 per experimental group) were intravenously infected with 5 x 10(4) and 5 x 10(5) viable cells of Listeria monocytogenes SLCC 4013. About 50-80 h after this challenge, all mice of the untreated control groups succumbed to their infection. Pretreatment of experimental animals on the optimal immunoactive schedule with the galactoside-specific mistletoe lectin (ML-1; 1 ng/kg body weight; days 1, 4, 5, 6 before challenge), however, evidently reduced the lethality of listerial infection (survival rate 60%).

Murine thymocyte proliferation, maturation and emigration in response to selenium.

Systemic (intraperitoneal) administration of sodium selenite (CAS 26970-82-1, Se) for 7 subsequent days (0.1 or 0.2 micrograms Se daily per 20 g of body weight; these concentrations were calculated from recommendations in human medicine) into BALB/c-mice could be shown to induce thymocyte proliferation and maturation. The increase in thymocyte numbers per mg organ weight was most pronounced after administration of the higher Se concentration. Determination of thymic lymphatic subsets revealed considerable up-regulations of mature T-cells expressing helper/inducer (L3T4) phenotype and immature cells expressing both (L3T4/Lyt-2) antigens. Thus, administration of Se (0.2 micrograms per mouse for a defined period) accelerated murine thymocyte proliferation and maturation. Counts of BALB/c-mice peripheral blood lymphocytes (PBL) revealed statistically significant increases after Se administration. The determination of activated PBL expressing interleukin (IL)-2 receptors proved that administration of Se induced a potent immunostimulation since these cells were significantly enhanced.

A new set of regulatory molecules in plants: A plant phospholipid similar to platelet-activating factor stimulates protein kinase and proton-translocating ATPase in membrane vesicles.

1-O-alkyl-2-acetyl-sn-glycero-3-phosphocholine, an ether phospholipid from mammals known as platelet-activating factor (PAF), specifically stimulates proton transport in zucchini (Cucurbita pepo L.) microsomes (G.F.E. Scherer, 1985, Biochem. Biophys. Res. Commm. 133, 1160-1167). When plant lipids were analyzed by two-dimensional thin-layer chromatography a lipid was found with chromatographic properties very similar to the PAF (G.F.E. Scherer and B. Stoffel, 1987, Planta, 172, 127-130). This lipid was isolated from zucchini hypocotyls, red beet root, lupin root, maize seedlings and crude soybean phospholipids. It had biological activity similar to that of the PAF, based on phosphorus content, and stimulated the steady-state ΔpH in zucchini hypocotyl microsomes about twofold. Other phospholipids, monoglyceride, diglyceride, triglyceride, oleic acid, phorbol ester, and 1-O-alkylglycerol did not stimulate proton transport. When microsomes were washed the PAF was ineffective but when soluble protein was added the PAF stimulation of H(+) transport was reconstituted. The soluble protein responsible for the PAF-dependent stimulation of transport activity could be partially purified by diethylaminoethyl Sephacel column chromatography. In the same fractions where the PAF-dependent transport-stimulatory protien was found, a protein kinase was active. This protein kinase was stimulated twofold either by the PAF or by Ca(2+). When Ca(2+) was present the PAF did not stimulate protein-kinase activity. When either the PAF, protein kinase, or both were added to membranes isolated on a linear sucrose gradient, ATPase activity was stimulated up to 30%. Comparison with marker enzymes indicated the possibility that tonoplast and plasma-membrane H(+)-ATPase might be stimulated by the PAF and protein kinase. We speculate that a PAF-dependent protein kinase is involved in the regulation of proton transport in plants in vitro and in vivo.

[Immunoactive effects of various mistletoe lectin-1 dosages in mammary carcinoma patients]. / Immunaktive Wirkung verschiedener Mistellektin-1-Dosierungen in Mammakarzinom-Patientinnen.

Cellular aspects of the immunomodulating activity of a proprietary mistletoe extract (Eurixor) standardized for mistletoe lectin-1 (ML-1) were investigated in patients suffering from mammary carcinoma (n = 20). Regular subcutaneous injections of the different dosages (0.5 and 1.0 ng ML-1/kg body weight, twice a week, for 5 weeks) yielded statistically significant increases of defined peripheral blood lymphocyte subsets (helper T-cells, natural killer (NK)-cells) which are generally believed to be involved in antitumor activity. Moreover, administration of either ML-1 concentration resulted in enhanced expression of activation markers such as interleukin-2 receptors and HLA/DR-antigens on peripheral blood T-lymphocytes. This study suggests that regular subcutaneous administration of both ML-1 concentrations (0.5 and 1.0 ng/kg body weight) can efficiently stimulate the cellular immune system of cancer patients.

Immunomodulating ability of galactoside-specific lectin standardized and depleted mistletoe extract.

Commercially available mistletoe extract standardized for the galactoside-specific lectin (ML-1; Eurixor) and a chromatographically ML-1-depleted preparation (same charge no. and composition of remaining components) were tested for their immunomodulating potency. In BALB/c-mice, regular subcutaneous administration of the optimal immunomodulating dosage (1 ng ML-1/kg body weight) could be shown to induce no influence on spleen weight, a non-significant increase of thymus weight, a significant increase of thymocyte, peritoneal macrophage, peripheral blood leukocyte, lymphocyte and monocyte and monocyte counts, and a significant decrease of peripheral blood granulocyte counts. Administration of analogue volumes (concentrations) of ML-1-depleted extract, however, did not induce any immunopotentiation. Accordingly, it may be assumed, that the galactoside-specific lectin (ML-1) represents the main immunomodulating component in commercially available mistletoe extracts.

Accumulation of polyunsaturated fatty acids by concentrate selecting ruminants.

Depot fat samples from ruminants of different feeding type and--for comparison--fat samples from simple-stomached animals were collected within 3 months. Individual fatty acid proportions, especially the relation of essential, polyunsaturated fatty acids to nonessential, saturated plus monounsaturated fatty acids were analyzed by gas chromatography. Species can be separated into two distinct groups: Depot fat of wild boar showed by far the highest content of essential fatty acids compared with all ruminant species. The subsequent inter-ruminant comparison yielded a further separation into two distinct groups related to feeding type. Roe deer and moose, constituting the first group of concentrate selectors, showed significantly higher percentages of polyunsaturated fatty acids than the other ruminant species comprised of the grass and roughage eaters or intermediates. The data document that the mode of feeding and/or the diet affects the body composition of the species investigated and that the depot fat composition of these ruminant species is markedly related to feeding type.

Ceramide-independent CD28 and TCR signaling but reduced IL-2 secretion in T cells of acid sphingomyelinase-deficient mice.

Ceramide generated by lysosomal acid sphingomyelinase (aSMase) has been proposed to contribute to CD28 co-stimulatory signaling pathways. We used an aSMase-deficient mouse line (asmase-/-) to elucidate the role of the aSMase in splenocytes stimulated with either a combination of anti-CD3 and anti-CD28 antibodies, the lectin concanavalin A (Con A) or the superantigen staphylococcal enterotoxin B. All stimuli were shown to induce IL-2 expression, Con A additionally triggered the expression of high-affinity IL-2 receptor. However, in asmase-/- mice secretion of IL-2 was significantly reduced, whereas the intracellular IL-2 levels were elevated. Proliferation of anti-CD3/anti-CD28 or Con A-stimulated aSMase-deficient splenocytes was reduced up to 50% after 72 h in comparison to wild-type cells. We conclude that ceramide generated by aSMase is not involved in CD28 signal transduction, but rather a perturbation of the secretory system is responsible for the impaired proliferation of aSMase-deficient splenocytes.

Modulating activity of mistletoe lectins 1 and 2 on the lymphatic system in BALB/c-mice.

The galactoside-specific lectin (mistletoe lectin-1, VAA-1) and the N-acetylgalactosamine-specific lectin (mistletoe lectin-2, VAA-2) were purified from aqueous mistletoe extract and checked for their immunoactive potency. Regular subcutaneous administration of the optimal immunomodulating VAA-1 /VAA-2 dosage (1 ng lectin/kg body weight) could be shown to modulate thymocyte proliferation, maturation, emigration and activation in BALB/c-mice. Thus, the increase in thymocyte counts was statistically significant after VAA-1 treatment. However, analogue VAA-2 applications significantly decreased thymocyte proliferation. Determinations of lymphatic subsets revealed considerable upregulation (after VAA-1 treatment) and significant downregulation (after VAA-2 treatment) of immature L 3 T4(+)/Lyt-2(+) thymic cells. Counts of mature cells expressing helper/inducer (L3T4(+)) or suppressor/cytotoxic (Lyt-2(+)) phenotypes did not present remarkable differences after VAA-1/VAA-2 administration. Counts of BALB/c-mouse peripheral blood cells revealed evident (statistically non-significant) increases of lymphocytes (PBL) and monocytes (PBM) after VAA-1 treatment, however, cell counts after VAA-2 administration were comparable to non-treated control animals. The determination of activated PBL expressing interleukin IL-2 receptors proved that VAA-1 induced a potent immunostimulation, since these cells were significantly enhanced after VAA-1 administration but not after VAA-2 treatment.