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Cytotoxic effects of some animal and vegetable extracts and some chemicals on liver and colon carcinoma and myosarcoma.

OBJECTIVE: To study, the cytotoxic effects of some biological and chemical agents on G1, S, G2, M and G0 phases of liver and colon carcinomas and myosarcoma cells obtained with chemical carcinogens dimethylbenzanthracene (DMBA) and cadmium chloride. METHODS: Eight rabbit livers, colon carcinoma and myosarcoma cell lines were obtained by injection of DMBA in the Biology Laboratory, of the University of Dumlupinar, Kutahya, Turkey between January 2001 and June 2003. All lines were grown at 37 degrees celsius and 5% carbon dioxide in sterile RPMI-1640 medium with 10% fetal bovine serum after addition of glutamate, penicillin (50 units/ml) and streptomycin (50 ug/ml) (complete medium). Cells were grown on standard tissue culture plastic flasks to 80% confluence and passed by trypsinization. RESULTS: Tortoise (Testudo graeca) shell, sponge (Geodia cydonium), medusa (Aurelia aurita), meat flies (Calliphora erythrocephala) larva, frog (Rana ridibunda) larva and juniper (Juniperus communis) berry extracts killed a large amount of the liver and colon carcinomas and the myosarcoma cells in G2, M and G0 phases (p<0.01). The mistletoe (Viscum album) extract had more effect in only the G0 phase (p<0.05). Genistein, genistin, glycitein, glycitin, daitzein and daitzin have significantly decreased in the cancer cells tests, particularly, genistein and daitzein caused the apoptotic effect in G2, M and G0 phases (p<0.01). Cesium chloride, a mixture of cesium chloride with magnesium chloride had the most effect on tumor cells (p<0.01). AzhexSi, Azhex-AzhepSi, Et-Azhex-AzhepSi, AzhepSi, Hexamine and DL 54 have been inhibited in various levels of the cancer cells (p<0.05, p<0.01). CONCLUSION: This data suggest that some biological extracts and chemicals tested may be useful chemotherapeutic agents to inhibit the growth of cancer cells. This study sheds some light for new anti cancerogenic experiments preventing various cancers on humans.

Animals↗

Hemorrheologic changes induced by aqueous extract of Ligaria cuneifolia: treatment by two different administration routes.

Ligaria cuneifolia (R. et P.) Tiegh. (Loranthaceae) (Argentine mistletoe) is usually used in local folk medicine. This work focuses on the hemorrheologic parameters in the treatment with an aqueous extract of Ligaria cuneifolia (Lc) by two different administration routes: intraperitoneal (i.p.) and intravenous (i.v.). Adult male Wistar rats were injected by via i.p. or by via i.v. with: saline solution; 2.5 mg/100 g body weight of Lc and 5.5 mg/100 g body weight of Lc. The relative viscosity of blood (eta r)(45/Hct) was measured showing that Lc-treatment by via i.p. produced an increase of about 69% while Lc by via i.v. enhanced the parameter about 47%. All of Lc-treated animals showed a significant increase in the rigidity index (RI). The mean corpuscular hemoglobin concentration (MCHC) exhibited an increase of about 15% in all the treated groups. Lc-treatment by via i.p. produced a diminution of plasma cholesterol level associated with RI augmentation which induced an increase of (eta r)(45/Hct). By via i.v. Lc produces both RI and (eta r)(45/Hct) augmentation by increasing MCHC but without modifying plasma cholesterol level, indicating a direct Lc-action on the internal viscosity of the erythrocyte.

Animals↗

Influence of complementary Viscum album (Iscador) administration on microcirculation and immune system of ear, nose and throat carcinoma patients treated with radiation and chemotherapy.

With the techniques of vital microscopic and reflection spectrometric imaging, representative characteristics of microcirculation and immunology of white blood cells were evaluated before, during and after radiotherapy and chemotherapy of patients suffering from ear, nose and throat carcinomas. Adverse effects of radiotherapy and chemotherapy on the microcirculation and the immune system were decreased and reconstitution processes were accelerated by complementary administration of a standardized mistletoe extract (Iscador).

Aged↗

Reflections on cancer therapy.

A schedule for treatment of cancer patients with low-dose combined chemotherapy infusions is presented for discussion. This schedule is designed to activate the immune system. Active agents such as mistletoe preparations etc. which are at present still controversial, balneological applications, diet, ethics and nursing home facilities are mentioned. Finally the question is raised as to whether it may be appropriate under certain circumstances in the individual case to institute abortive cytostatic treatment even in suspicion of a cancerous condition, i.e. at a time when the organ diagnosis has not yet been substantiated.

Antineoplastic Combined Chemotherapy Protocols↗

[Why do cancer patients use alternative medicine?].

160 patients (53.3%) replied to an anonymous questionnaire distributed to 300 consecutive patients of our Outpatient Oncology Clinic. 83 patients (53%) mentioned some experience of one or more alternative methods of cancer treatment. Most often cited were various herbal teas (35 instances), beetroot juice (16), Vogel plant extracts (15), laying-on of hands (14), homeopathic medicine (13), the mistletoe extract Iscador (13), magnetopathy (12), various diets (10), acupuncture (10) and psychological methods (9). Resort to alternative methods of treatment correlated significantly with lower age (51.5 years vs 59.8 years, p = 0.001). The reasons for using alternative medicine were the desire to do everything possible to regain health (49), to use one's psychological forces as well (35), reports of successful cancer cures (28), desire for a holistic approach (23), hope of "softer" medicine with less severe side effects (18) and, in 7 cases only, disappointment with conventional university medicine. The major source of information was relatives and friends, not the mass media. The physician should be aware of the locally available alternative medicine options and be able to advise his patients accordingly. He should also recognize and give due consideration to the patient's underlying desire for better control of his disease and a more holistic approach to care.

Complementary Therapies↗

[Immune modulators in the treatment of malignant melanoma].

The experience gained to date with immunomodulators used as therapeutical and adjuvant measures in the treatment of cutaneous malignant melanoma is reviewed. In large studies no therapeutic effect has been shown for levamisole, DNCB and BCG preparations. The effectiveness of monoclonal antibodies, histamine type 2 receptor antagonists, thymus factors and mistletoe extracts cannot yet be assessed due to a lack of clinical studies. The recent use of the lymphokines: tumor-necrosis factor, interleukin 2 and interferons, has shown that tumor therapy with immunomodulators is possible, but many questions still remain to be answered, so that their use, especially as an adjuvant measure, should be restricted to controlled studies.

Adjuvants, Immunologic↗

[Are paramedical means and methods justified in the treatment of breast carcinoma?].

In industrial countries breast cancer is the second frequent malignancy of women. The often metastasizing (65 to 70%) and long-lasting illness leads patients in more than 50% to paramedical treatments. An analysis of the unproven methods and the remedies concerning the effectiveness against breast cancer is done: mistletoe preparations, nutrition directions, organotherapy (cell therapy, cytoplasmatic and thymus therapy), enzyme-preparations and further paramedical remedies and methods. In the last part, the discussion covers the findings of the 'Gesellschaft für Biologische Krebsabwehr', which presumes that biological remedies will lead to a turn in cancer treatment. An exact analysis of the paramedical literature shows that the quoted successes of paramedical treatment are nearly parallel to the successes of 'ortho-medical' therapies, which are not mentioned at all or only on incidentally.

Anthroposophy↗

Histamine receptor-bearing lymphocytes (HRL). VIII. Splenopentin-accelerated reconstitution of the immunoresponsiveness after elimination of HRL.

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.

Animals↗

[Characterization of lectin receptors on cell surfaces with regard to functional aspects].

The actual definitions of lectins are presented and discussed and lectins are classified as one group of the affinitins. The ability of lectins to bind carbohydrates serves as a widely used principle in the nature for cell recognition and adhesion processes. By means of labelled glycoconjugates the identification, localization and quantitation of receptor bound lectins can be investigated as by lectinological as by a combination of lectinological and immunological reactions. The special properties of toxic lectins are discussed by use of the lectin I from mistletoe as characteristic example. The coupling of A-chains with antibodies (immunotoxins) or other haptomers (affinotoxins) leads to agents render it possible to influence directly cell functions.

Animals↗

[Value of the lymphocyte transformation test for determining immune status].

Studies on lymphocyte stimulation in vitro were carried out with various perspectives. Apart from the well-known mitogens such as PHA and ConA, Candida extract and mercury ions had a relatively strong stimulatory effect, mistletoe extract a moderate one, nickel ions and trichophytin none at all. In malignant lymphomas the degree of stimulation was greatly reduced. In malignant melanomas the degree of stimulation, which at the beginning was often quite low, increased during the course of treatment. There was no correlation between the response to recall antigens and the degree of lymphocyte stimulation. The lymphocyte stimulation test did not prove reliable for the detection of drug allergies.

Drug Hypersensitivity↗

Immunological reactivity of tumor associated glycoproteins prepared by affinity chromatography.

Crude 3M KCl extracts prepared from tumor tissue and ascitic tumor cells of methylnitrosourea-induced fibrosarcoma of CBA mice or of a human stomach adenocarcinoma were gelfiltrated on Ultrogel AcA 34. Single fractions of these preparations were incubated with spleen cells of tumor bearing mice or blood peripheral lymphocytes. The response was detected by MEM-technique. Antigenic activities were found in molecular weight ranges from 200 to 300 kD (P1) and from 40 to 70 kD (P2). Single glycoproteins of the P1- and P2-fractions could be separated by affinity chromatography on immobilized D-galactose specific mistletoe lectin I, as detected by electrophoresis on microgradient gels (1.2-40%) or on SDS polyacrylamide slab gels. Especially the isolated glycoproteins of the P1-fractions are responsible for the observed antigenic reactivity, assayed in vitro by the MEM-technique and in vivo by foot pad swelling test.

Adenocarcinoma↗

[Unconventional therapeutic methods in superficial bladder cancer].

Several unconventional agents or methods are used for recurrence prophylaxis of superficial bladder cancer. In animal experiments KLH (keyhole limpet hemocyanin) has shown an effect comparable to that of BCG on bladder carcinoma, resulting in an increase of natural killer cell activity. The few clinical data dealing with KLH are contradictory. A prospective randomized study with a large number of patients uniform high dosage and early start of instillation has not yet been performed for definitive evaluation of the clinical role of KLH. Whether mistletoe extracts or intravesical antineoplastic iontophoresis can prevent recurrence is not yet known. Vitamin A and megadose multivitamins in combination with intravesical BCG significantly decrease the recurrence rate. The most effective vitamin or the most important combination has not yet been identified, and the precise mechanism of action is also unknown.

Adjuvants, Immunologic↗

Phenotype-associated lectin-binding profiles of normal and transformed blood cells: a comparative analysis of mannose- and galactose-binding lectins from plants and human serum/placenta.

Surface glycoconjugates of normal and transformed blood cells are commonly characterized by plant lectins. To infer physiological significance of protein-carbohydrate interactions, mammalian lectins are obviously preferable as research tools. So far, human serum lectins have not been used to assess their binding to immunophenotyped human normal or transformed blood cells. Thus, our study combines two groups of lectins with different specificity from plant and human sources. Besides concanavalin A (ConA) we have isolated the mannose-binding protein and serum amyloid P component from human serum. Especially the mannose-binding protein is believed to play a role in host defence against bacteria and yeast cells with unknown impact on normal and tumor cells. These three lectins establish the first group. In addition to the immunomodulatory mistletoe lectin, whose binding can elicit enhanced cytokine secretion from mononuclear blood cells, we included the beta-galactoside-binding lectin (14 kDa) from human placenta in the second group. The initial series of measurements was undertaken using two-color flow cytometry to determine the phenotype-associated binding (based on cluster designation; CD) of the lectins to blood and bone marrow cells from normal donors and the cell line CEM (T-lymphoblastoid), KG1-A (primitive myeloid leukemia) and Croco II (B-lymphoblastoid). Heterogeneity was apparent for each lectin in the CD-defined cell populations. Significant differences in binding were noted between Viscum album agglutinin (VAA) and other lectins for CD4+ cells from blood and between mannose-binding protein (MBP) and VAA versus 14 kDa, ConA and serum amyloid P component (SAP) for CD19+ cells from bone marrow.(ABSTRACT TRUNCATED AT 250 WORDS)

Blood Cells↗

Quantitation and histochemical localization of galectin-1 and galectin-1-reactive glycoconjugates in fetal development of bovine organs.

The display of cellular oligosaccharide chains is known to undergo marked developmental changes, as monitored histochemically with plant lectins. In conjunction with endogenous lectins respective ligand structures may have a functional role during fetal development. The assumption of a recognitive, functionally productive interplay prompts the study of the expression of a tissue lectin and of lectin-reactive glycoconjugates concomitantly. Focusing on common beta-galactosides as constituents of oligosaccharide chains and the predominant member of the family of galectins in mammals, namely galectin-1, the question therefore is addressed as to whether expression of lectin and lectin-reactive glycoconjugates exhibits alterations, assessed in three morphologically defined fetal stages and in adult bovine organs. Using a sandwich ELISA, the level of the rather ubiquitous galectin-1 is mostly increased in adult organs relative to respective fetal stages, except for the case of kidney. This developmental course is seen rather seldom, when the amounts of lectin-reactive glycoproteins or glycolipids are quantitated in solid-phase assays after tissue homogenization. Western blotting, combined with probing by labeled galectin-1, discloses primarily quantitative changes in the reactivity of individual glycoproteins. Performing the same assays on extract aliquots with a plant agglutinin, namely the galactoside-binding mistletoe lectin, whose fine specificity is different from galectin-1, its reduced extent of binding in solid-phase assays and the disparate profile of lectin-reactive glycoproteins reveal a non-uniform developmental alteration within the group of structural variants of beta-galactosides. Although sample preparation can affect ligand preservation and/or presentation and thus restricts the comparability of biochemical and histochemical results, especially for soluble reactants, the histochemical studies on frozen and paraffin-embedded sections of bovine heart, kidney and liver demonstrate that the localization of the galectin and of lectin-reactive epitopes can show a similar distribution, as seen in liver and heart, with organ-typical quantitative changes of a rather similar staining profile (heart, kidney) or notable changes in the spatial distribution (liver) in the course of development. This report emphasizes the potential value of combined monitoring of the lectin and its potential in vivo ligands to contribute to eventually unravel organ-related function(s) of a tissue lectin.

Animals↗

Differential binding properties of Gal/GalNAc specific lectins available for characterization of glycoreceptors.

Differentiating the binding properties of applied lectins should facilitate the selection of lectins for characterization of glycoreceptors on the cell surface. Based on the binding specificities studied by inhibition assays of lectin-glycan interactions, over twenty Gal and/or GalNAc specific lectins have been divided into eight groups according to their specificity for structural units (lectin determinants), which are the disaccharide as all or part of the determinants and of GalNAc alpha 1-->Ser (Thr) of the peptide chain. A scheme of codes for lectin determinants is illustrated as follows: (1) F (GalNAc alpha 1-->3GalNAc), Forssman specific disaccharide--Dolichos biflorus (DBL), Helix pomatia (HPL) and Wistaria floribunda (WFL) lectins. (2) A (GalNAc alpha 1-->3 Gal), blood group A specific disaccharide--Codium fragile subspecies tomentosoides (CFT), Soy bean (SBL), Vicia villosa-A4 (VVL-A4), and Wistaria floribunda (WFL) lectins. (3) Tn (GalNAc alpha 1-->Ser (Thr) of the protein core)--Vicia villosa B4 (VVL-B4), Salvia sclarea (SSL), Maclura pomifera (MPL), Bauhinia purpurea alba (BPL) and Artocarpus integrifolia (Jacalin, AIL). (4) T (Gal beta 1-->3GalNAc), the mucin type sugar sequences on the human erythrocyte membrane(T alpha), T antigen or the disaccharides at the terminal nonreducing end of gangliosides (T beta)--Peanut (PNA), Bauhinia purpurea alba (BPL), Maclura pomifera (MPL), Sophora japonica (SJL), Artocarpus lakoocha (Artocarpin) lectins and Abrus precatorius agglutinin (APA).(5) I and II (Gal beta 1-->3(4)GlcNAc)--the disaccharide residue at the nonreducing end of the carbohydrate chains derived from either N- or O-glycosidic linkage--Ricinus communis agglutinin (RCA1), Datura stramonium (TAL, Thorn apple), Erythrina cristagalli (ECL, Coral tree), and Geodia cydonium (GCL). (6) B (Gal alpha 1-->3Gal), human blood group B specific disaccharide--Griffonia(Banderiaea) simplicifolia B4 (GSI-B4). (7) E (Gal alpha 1-->4Gal), receptors for pathogenic E. coli agglutinin, Shiga toxin and Mistletoe toxic lectin-I (ML-I) and abrin-a.

Acetylgalactosamine↗

Modification of the functional activity of neutrophils treated with acute phase response proteins.

The effects of alpha1-acidic glycoprotein (AGP) and its subfractions (glycoforms) with different affinity for concanavalin A on generation of H2O2 by human neutrophils exposed to stimulators of different nature, namely, galactose-specific mistletoe (Viscum album) lectin (VAA), digitonin, and N-formyl-Met-Leu-Phe, a chemotactic peptide (FMLP), were studied. Within the concentration range of 13-500 microgram/ml, AGP and its glycoforms produced dose-dependent inhibition of digitonin-induced cell responses. AGP also inhibited the VAA-induced generation of H2O2; however, this cell response was potentiated by low concentrations (50 microgram/ml) of AGP and AGP-A. FMLP induced the most consistent response of neutrophils, which changed only slightly in the presence of AGP and AGP-B; however, low concentrations of AGP-A inhibited this response. The presence of sialic acid in the terminal position of carbohydrate antennae of AGP is not necessary for its inhibitory effect on human neutrophil respiratory burst because asialo-AGP (250 microgram/ml) inhibited H2O2 generation by cells stimulated with agonists of the NADPH-oxidase system of phagocytes. In contrast to AGP, two other acute phase response proteins displaying a lectin activity (C-reactive protein and serum amyloid P component) within the concentration range of 10-100 microgram/ml produced no significant effect on H2O2 generation by stimulated neutrophils. These data suggest that AGP is an effector molecule responsible for feedback regulation of the functional activity of neutrophils.

C-Reactive Protein↗