Electronic Library of Scientific Literature


NEOPLASMA


Volume 50 / No. 4 / 2003


Preliminary identification of carotenoids in malignant and benign neoplasms of the breast and surrounding fatty tissue.

Czeczuga-Semeniuk E, Wolczynski S, Markiewicz W.

Department of Gynecological Endocrinology, Medical Academy of Bialystok, Bialystok, 15-276 Poland. czeczuga@wp.pl

An attempt has been made to identify the carotenoids present in the tissue of neoplastic tumors and the surrounding fatty tissue taken from women with histologically diagnosed cancer (ca ductale infiltrans, G2,G3; n=20) and those with benign changes (fibroadenoma, n=20). Carotenoid pigments were isolated using column and thin-layer chromatography. Prior to chromatography, the material was homogenized with acetone under nitrogen in dark glass bottles and the extracts kept in a refrigerator until analyzed. In the present study, we isolated 13 carotenoids belonging to provitamin A and nonprovitamin A carotenoids. The total content of carotenoids in microg/g of tissue was slightly lower in cancers and the surrounding fatty tissues in comparison to benign changes, but in general it was higher in the fatty tissue surrounding the tumors, irrespective of their histological structure (the mean values for cancers 20.433+/-10.64 vs fatty tissue 25.361+/-12.025, p<0.01; and the mean values for benign changes 22.889+/-12.011 vs fatty tissue 27.021+/-13.180, p<0.01). Epoxide carotenoids - lutein epoxide and violaxanthin, were predominant in fatty tissue, both in malignant and benign changes; epoxide carotenoids - mutatoxanthin and lutein epoxide and other carotenoids such as zeaxanthin, canthaxanthin, lutein and neoxanthin were predominant in neoplastic material. Beta carotene and lutein epoxide were found in all samples, alpha carotene was found in 50% of them. Antheraxanthin was present in fatty tissue only. Beta carotene, the main provitamin A carotenoid, content in the material examined ranged from 2.43 to 4.33% in tumor tissue and in fatty tissue surrounding the tumors it was twice as higs. Such carotenoids as 3'-lutein, canthaxanthin and astaxanthin were sporadic. No reoccurring carotenoid "sequences" were found despite the same histopathological diagnosis. No relationship was found between the neoplasm histopatological grade, lesion diameter and the occurrence of specific carotenoids.

Neoplasma. 2003; 50(4): 280-286.


Androgen receptor in prostate carcinoma: immunohistochemical and ligand saturation analyses.

Vagundova M, Vagunda V, Vermousek I, Rovny A.

Masaryk Memorial Cancer Institute, 656 53 Brno, Czech Republic. vagundov@mou.cz

Status of androgen receptor (AR) in prostate carcinoma is biologically important. Therefore, more methods assessing AR abnormalities are warranted. Immunohistochemical (IHC) and ligand saturation (LSA) assays were not compared in details. AR in 53 cases were tested by monoclonal antibody (Ab) F39.4.1 (Biogenex), polyclonal Ab N-20 (Santa Cruz) and by ligand saturation analysis with (3)H-methyltrienolon. Statistical analyses were performed with Spearman's nonparametric rank test including neoadjuvant therapy subgroups treated by antiandrogens, combined androgen blockade (22 cases; flutamide with gosereline) or without therapy. By using monoclonal Ab we found AR positive tumor nuclei in 46 cases. Mean of positives was 64%, median 75%. The polyclonal Ab was not sufficiently specific. With LSA AR were found in 43 cases. Mean level was 6.6 fmol/mg, median 5.5 fmol/mg. Comparing IHC with LSA, we noted correlation trend only for the monoclonal Ab (r=0.35; p=0.02). With thresholds 70% positive nuclei for IHC and 6 fmol/mg for LSA, there were 66% and 43% cases positive with IHC and LSA, respectively. The LSA and IHC positives did not show significant agreement, concordance level being 58% only. We found significant IHC-LSA correlation (r=0.68; p=0.004) solely in combined androgen blockade subgroup with 82% level concordance. Our study has demonstrated that AR IHC and LSA are independent complementary methods. Significant correlation between LSA and IHC show only cases treated with combined androgen blockade. An explanation hypothesis is discussed concerning LH-RH influence on free AR capable of ligand binding. IHC as well as LSA have specific biologic significance and may be useful for prostate cancer diagnostic and therapy.

Neoplasma. 2003; 50(4): 287-290.


The free radical scavenging activity of four flavonoids determined by the comet assay.

Horvathova K, Novotny L, Vachalkova A.

Cancer Research Institute, Slovak Academy of Science, 833 91 Bratislava, Slovak Republic. katarina.horvathova@savba.sk

Flavonoids are reported to exhibit a wide variety of biological effects, including antioxidant and free radical scavenging activities. The aim of our study was to determine the cytotoxicity of flavonoids quercetin, rutin, apigenin and luteolin and their ability to protect DNA molecules against H2O2-induced damage. Cytotoxicity of studied flavonoids was tested in murine leukemia L1210 cells by the trypan blue exclusion technique. DNA strand breaks were determined using the alkaline single-cell gel electrophoresis (comet assay). Quercetin was found to possess the highest protective effect among the flavonoids studied (45%).The protective activity determined was lower for luteolin (40%). Protective effect of apigenin (600 microM/L) was only marginal (2%). However, at the higher concentration of apigenin (1200 microM/L), this flavonoid induced DNA single strand breaks. This indicates the ability of apigenin to serve as a pro-oxidant. Rutin had no protective effect on DNA single strand breaks induced by H2O2.

Neoplasma. 2003; 50(4): 291-295.


Clinical-scale selective depletion of alloreactive T cells using an anti-CD25 immunotoxin.

Michalek J, Collins RH, Vitetta ES.

Cancer Immunobiology Center, Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas, TX 75390 USA. jmichal@med.muni.cz

Allogeneic hematopoietic stem cell transplantation is the treatment of choice for many hematological malignancies. Its efficacy is limited by graft-versus-host disease (GVHD), the leading cause of post-transplant morbidity and mortality. GVHD is mediated by a subpopulation of T cells in the stem cell graft. Ex vivo T cell depletion of all T cells of the graft can prevent development of GVHD but can lead to a delay in immune reconstitution and an increase of potentially lethal opportunistic infections and leukemic relapses. Hypothetically, an approach that enables a selective depletion of the alloreactive donor T cells that cause GVHD while preserving third party (anti-leukemic and anti-microbial) reactivity would be optimal for recipients of HSCT. Our preliminary data demonstrated that an anti-CD25 immunotoxin, which reacts with a cell surface activation antigen, can selectively deplete alloreactive donor T cells activated by non-leukemic recipient white blood cells while preserving the beneficial third-party reactivity in vitro. In this report we describe a method for clinical-scale ex vivo selective depletion of alloreactive donor T cells using the anti-CD25 immunotoxin, RFT5-SMPT-dgRTA. Two logs of alloreactive T cells could be selectively depleted while preserving third party reactivity. This method was reproducible in 10 pre-clinical experiments with 8 HLA-mismatched healthy volunteer pairs and 2 HLA-matched sibling donor/patient pairs.

Neoplasma. 2003; 50(4): 296-269.


Ethanol fixation of lymphoma samples as an alternative approach for preservation of the nucleic acids.

Soukup J, Krskova L, Hilska I, Kodet R.

Laboratory of Molecular Pathology, Department of Pathology and Molecular Medicine, Charles University, 2nd Medical School, Prague 5 - Motol, 150 06 Czech Republic.

Molecular methods play an important role in diagnostic pathology of lymphomas. PCR based demonstration of clonality or detection of a specific chromosomal translocation may determine the exact classification of the lymphoma. Hence the final diagnosis may depend on the quality of preserved nucleic acids in the bioptic specimen. The integrity of DNA and RNA may be damaged by formalin fixation, which destroys the nucleic acids by fragmentation. Therefore, a portion of each lymphoma sample should be frozen. To substitute freezing techniques we utilized ethanol as a fixative, which preserves nucleic acids. We compared PCR and RT-PCR products from lymphoma samples, which were differently pre-treated by ethanol fixation, formalin fixation and freezing. The ethanol fixed samples retained a high quality of both DNA and RNA and provided reproducible PCR products similar to frozen samples and significantly better then those extracted from formalin fixed samples. We may recommend ethanol as a complementary fixative for all pathology laboratories where deep freezing in not routinely available.

Neoplasma. 2003; 50(4): 300-304.


Antimelanoma immunity in vitiligo and melanoma patients.

Juranic ZD, Stanojevic-Bakic N, Zizak Z, Babovic N, Radovic-Kovacevic V, Stanojkovic T, Dzodic R.

Institute for Oncology and Radiology of Serbia, Belgrade, Yugoslavia. juranicz@ncrc.ac.yu

Cutaneous melanoma and vitiligo are diseases etiology of which evolves around melanocytes. The nature of immunological disturbances associated with these diseases is not elucidated. The experiments performed in this work were aimed to determine antimelanoma immunotoxicity in patients with melanoma and patients with vitiligo. Twelve patients with melanoma, ten patients with vitiligo and seventeen healthy volunteers were studied. The cytotoxicity of PBMC was evaluated indirectly through determination of target melanoma (Fem-x) or control tumor (HeLa) cell survival, in the presence of 15% of AB or autologous sera, by MTT test. The mean values of antimelanoma cytotoxicity in AB serum were similar in both patients groups and in controls. However, the frequency of patients with the enhanced cytotoxicity against melanoma cells, in relation to control tumor cells, was lower in both patients groups than in controls. The intensity of antimelanoma cell-mediated cytotoxicity in melanoma patients, in the presence of autologous serum, was significantly lower in comparison to that found in control subjects and vitiligo patients (p<0.014, in both cases). This indicates that some factors from melanoma patient's sera contribute to impairment of the cytotoxicity of autologous PBMC, while other factors from the serum of vitiligo patients and control subjects enhanced their PBMC antimelanoma cytotoxicity.

Neoplasma. 2003; 50(4): 305-309.


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