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FOLIA HISTOCHEMICA
ET CYTOBIOLOGICA
Vol. 43, No. 2, 2005
CONTENTS
A. Janeczko, A. Skoczowski: Mammalian sex hormones in plants pp. 71-79
A. Szuster, M. Kosz-Vnenchak: Autocrine growth
regulation of W12 and GCA cells in culture pp. 91-102
ABSTRACTS
Mammalian sex hormones in plants
Anna Janeczko and Andrzej Skoczowski
Institute of Plant Physiology, Polish Academy of Sciences, Kraków,
Poland
Abstract: The occurrence of mammalian sex hormones and their physiological role in plants is reviewed. These hormones, such as 17b-estradiol, androsterone, testosterone or progesterone, were present in 60-80% of the plant species investigated. Enzymes responsible for their biosynthesis and conversion were also found in plants. Treatment of the plants with sex hormones or their precursors influenced plant development: cell divisions, root and shoot growth, embryo growth, flowering, pollen tube growth and callus proliferation. The regulatory abilities of mammalian sex hormones in plants makes possible their use in practice, especially in plant in vitro culture.
Authors e-mail: ania@belanna.strefa.pl
Characterization of human pituitary adenomas in cell cultures by light and electron microscopic morphology and immunolabeling
Ilona Fazekas1, Balázs Hegedûs1, Ernġ Bácsy2, Edit Kerekes1, Feliícia Slowik1, Katalin Bálint1 and Emil Pásztor1
1National Institute of Neurosurgery and 2Institute of Experimental Medicine, Hungarian Academy of Sciences, Budapest, Hungary
Abstract: The morphology and hormone production of pituitary adenoma cell cultures were compared in order to highlight their characteristic in vitro features. Cell suspensions were prepared from 494 surgical specimens. The 319 viable monolayer cultures were analyzed in details by light microscopy and immunocytochemistry within two weeks of cultivation. Some cultures were further characterized by scanning, transmission and immunogold electron microscopy. The viability and detailed in vitro morphology of adenoma cells wereas found to be characteristic for the various types of pituitary tumors. The sparsely granulated growth hormone, the corticotroph and the acidophil stem cell adenomas provided the highest ratio of viable cultures. Occasionally, prolonged maintenance of cells resulted in long-term cultures. Furthermore, a variety of particular distributions of different hormone-containing granules were found in several cases. Both light microscopic and ultrastructural analyses proved that the primary cultures of adenoma cells retain their physiological features during in vitro cultivations. Our in vitro findings correlated with the routine histopathological examination. These results prove that monolayer cultures of pituitary adenoma cells can contribute to the correct diagnosis and are valid model systems for various oncological and neuroendocrinological studies.
Authors e-mail: hegedusb@missouri.edu
Abstract: Two rat kidney cell lines transformed by two strains of ASV virus were investigated. It was demonstrated that these two lines (1) showed density-independent growth, (2) had a decreased requirement for serum in the culture medium, (3) had the ability to grow in a chemically defined medium (without serum), and the rate of this growth had increased with the increase in starting density of cells, and (4) had the ability of anchorage-independent growth, even without serum. These results confirmed autostimulation of growth of W12 and GCA cells. It was also shown that the crude conditioned media contained autocrine growth factors, which could be extracted with 1M acetic acid. The extracts (AEs) stimulated the growth of the parental cells and NRK-49F cells almost as well as 5% calf serum and the extraction resulted in several-fold purification of mitogenic substances. These substances were not only specific to parental lines, but also stimulated growth of other transformed lines and normal NRK-49F cells. Extracts from the conditioned media of W12 and GCA cells intensified the rate of anchorage-independent growth in the concentration-dependent manner. In AE-W12, two peaks of mitogenic activity were detected (F1, F2) and similarly in AE-GCA (F3, F4). Fractions F2 (~ 8 kDa), F3 (~25 kDa) and F4 (~ 12 kDa) were thermostable but F1 (~ 45 kDa) was thermolabile. All four fractions were sensitive to trypsin and DTT treatment, and were acid-stable. Using ELISA kit it was shown that W12 and GCA cells released TGFb 1 and GCA cells released very small quantities of bFGF. These results confirmed the autocrine regulation of growth in both cell lines.
Authors e-mail: szuster@awe.mol.uj.edu.pl
The effects of synthetic salmon calcitonin on thyroid C and follicular cells in adult female rats
M. Sekuliĉ, B. oiĉ-Jurjeviĉ, B. Filipoviĉ, V. Miloeviĉ, N. Nestoroviĉ and M. Manojloviĉ-Stojanoski
Institute for Biological Research Sinia Stankoviĉ, Belgrade, Serbia and Montenegro
Abstract: Structural and morphometric features of thyroid C and follicular cells were studied in adult rat females after treatment with synthetic salmon calcitonin (CT). The animals were chronically treated with either a low (10 IU/kg b.w) or a high (100 IU/kg b.w) dose of CT. A stereological method was applied to determine the volume density and the number of immunoreactive C cells. The height and volume density of follicular epithelium, colloid, interstitium and the follicles (epithelium plus colloid), as well as the index of activation rate were calculated. A significant decrease in body weight, as well as the volume density of immunoreactive C cells and the number of C cells per mm2, was observed in rats treated with both doses of CT. The height and volume density of follicular epithelium and follicles, as well as the index of activation rate were significantly increased in the animals given the high CT dose, while the volume densities of colloid and interstitium were reduced. No significant changes in the examined morphometric parameters were detected after treatment with the low CT dose. According to these results it can be concluded that the structural features of thyroid C and follicular cells were affected by the high dose CT treatment in the opposite manner, while the low dose CT treatment influenced only C cells.
Author's e-mail: sekmic@ibiss.bg.ac.yu
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Maspin and c-erbB-2 expression in correlation with microvessel density in invasive ductal breast cancer
Miros³aw Sopel1, Izabela Kasprzyk2 and Izabela Berdowska3
1Department of Histology and Embryology, 2Department of Anatomy, and 3Department of Medical Biochemistry, Medical University, Wroc³aw, Poland
Abstract: Maspin is a unique member of the serpin family involved in regulation of cell migration, apoptosis and angiogenesis in breast and prostate cancers. In this study maspin expression in comparison with c-erbB-2 (HER2/neu) oncogene expression and microvessel density was investigated. The examined material included specimens of primary invasive ductal breast cancer derived from 69 patients. They were analyzed immunocytochemically to assess maspin and c-erbB-2 expression, as well as microvessel density using endothelium marker CD31. In the studied cancers, maspin expression in cancer cells was detected in more than half of the cases (50.73%). Although statistically insignificant (p=0.27), maspin expression showed decreasing tendency with the increase of tumor grade. C-erbB-2 oncogene expression was observed in 78.26% of the examined cancers. Statistically significant positive correlation was found between c-erbB-2 expression and tumor grade (p<0.005). Analysis of the dependence between maspin and c-erbB-2 expression exhibited statistically significant inverse correlation (p<0,001). Mean microvessel density (MVD) of the studied cancers was 71.64 (SD=19.36). MVD decreased with the increase of maspin expression, whereas in the cases showing c-erbB-2 overexpression MVD was clearly higher. Both correlations were statistically significant (p<0.005). In conclusion, it could be stated that increase in maspin expression is associated with weaker expression of c-erbB-2 oncogene and lower microvessel density, which implies a significant role of maspin in tumor biology. However, the exact mechanism of maspin action (including its potential role in angiogenesis), as well as the assessment of its prognostic significance in breast cancer require further studies.
Authors e-mail: misopel@hist.am.wroc.pl
Flow cytometric analysis of CD55 and CD59 expression on blood cells in paroxysmal nocturnal haemoglobinuria
Grzegorz Dworacki1, Jan Sikora1, Ewa Mizera-Nyczak1, Magdalena Trybus1, Iwona Mozer-Lisewska2, Anna Czyż3 and Jan Żeromski1
Departments of 1Clinical Immunology, 2Child Infectious Diseases and 3Haematological Haemopoietic Proliferative Diseases, University of Medical Sciences, Poznañ, Poland
Abstract: PNH is a rare clonal disorder of hematopoietic stem cells, therefore all blood cells lineages are involved. The main feature is an increased sensitivity of erythrocytes to complement-mediated cell lysis due to deficiency of membrane-bound GPI (glycosylphosphatidylinositol)-anchored proteins which normally function as inhibitors of reactive hemolysis. In the present study, we performed flow cytometric analysis using monoclonal antibodies against CD55 and CD59 for the detection of PNH-type clone in the blood of 50 patients (28 females and 22 males, age range 7-67 yrs). In one patient only we found a large population (95%) of granulocytes with decreased expression of both CD55 and CD59 molecules (type I PNH) and in two others with partial loss of CD55 expression (type II PNH). The expression was determined chiefly on granulocytes which in the control group showed reliable and high expression of CD55 and CD59.
Authors e-mail: gdwrck@wp.pl