K. Boczoń, E. Wandurska-Nowak, A. Wierzbicki, M. Frydrychowicz, I. Mozer-Lisewska, J. Żeromski: mRNA expression and immunohistochemical localization of inducible nitric oxide synthase (NOS-2) in the muscular niche of Trichinella spiralis pp. 209-213
E. Czeczuga-Semeniuk, S. Wołczyński, M. Dąbrowska, J. Dzięcioł, T. Anchim: The effect of doxorubicin and retinoids on proliferation, necrosis and apoptosis in MCF-7 breast cancer cells pp. 221-227
M. Rzeszutko, W. Rzeszutko, P. Dzięgiel: The morphological analysis of vasculature in thyroid tumours: immunoexpression of CD34 antigen pp. 235-240
M. Pawlikowski, A. Gruszka, M. Radek, J. Kunert-Radek: Chromogranin A in pituitary adenomas: immunohistochemical detection and plasma concentrations pp. 245-247
G. Hoser, D. Wasilewska, J. Domagała-Kulawik: Expression of Fas receptor on peripheral blood lymphocytes from patients with non-small cell lung cancer pp. 249-252
ABSTRACTS
Expression of connexins 26, 32 and 43 in the human colon – an immunohistochemical study
Luiza Kanczuga-Koda, Stanislaw Sulkowski, Mariusz Koda, Maria Sobaniec-Łotowska and Mariola Sulkowska
Department of Pathology, Medical University, Bialystok, Poland
Abstract: Gap junctional intercellular communication (GJIC) is a mechanism for direct cell-to-cell signalling and is mediated by gap junctions (GJs), which consist of proteins called connexins (Cxs). GJIC plays a critical role in tissue development and differentiation and is important in maintenance of tissue homeostasis. The purpose of the study was to evaluate the expression of Cx26, Cx32 and Cx43 in the human colon. Surgical specimens were obtained from patients who underwent surgical resection of colorectal tumours. Tissue samples (50 cases) were collected from normal colon, at the maximum distance from the tumor. Using antibodies for Cx26, Cx32 and Cx43, immunohistochemical detection was made. In epithelial cells, strong Cx26 immunoreactivity was found, whereas Cx32 and Cx43 were sparsely distributed. Strong Cx43 immunostaining in muscularis mucosae was observed. In the circular layer of muscularis externa, expression of Cx43 and Cx26 was seen, but only in the portion closest to the submucosa. No immunoreactivity was found in the longitudinal muscle layer. Small vessels stained positively only for Cx43. Furthermore, there was no difference in staining between samples derived from various sections of the colon. This study showed immunohistochemically for the first time the expression of Cx26 in human colon mucosa.
Author's e-mail: sulek@zeus.amb.edu.pl
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mRNA expression and immunohistochemical localization of inducible nitric oxide synthase (NOS-2) in the muscular niche of Trichinella spiralis
Krystyna Boczoń1, Elżbieta Wandurska-Nowak1, Andrzej Wierzbicki2, Magdalena Frydrychowicz3, Iwona Mozer-Lisewska4 and Jan Żeromski3
1Department of Biology and Medical Parasitology, 2Department of Biochemistry and Molecular Biology, 3Department of Clinical Immunology, 4Department of Child Infectious Diseases, University of Medical Sciences, Poznań, Poland
Abstract: The aim of this study was to demonstrate iNOS mRNA expression in muscular phase of experimental trichinellosis and to localize iNOS protein in T. spiralis-infected muscles using specific anti-iNOS monoclonal antibodies. The expression of iNOS mRNA in skeletal muscles from Trichinella spiralis-infected mice was examined using the reverse transcription PCR assay. Fragments of skeletal muscles were also subjected to the immunohistochemical reaction using specific anti-iNOS monoclonal antibodies followed by Dako-Ark test. mRNA for iNOS measured on day 21 after infection was expressed in the muscular phase of trichinellosis. Positive immunostaining for iNOS occurred in infiltrating mononuclear cells around the encapsulated larvae. iNOS-positive cells could be traced from the 21st day post infection (dpi); on 42 dpi and 90 dpi most cells expressed iNOS. By assessing expression of protein and its mRNA it can be concluded that iNOS is active in the pathology of skeletal muscle tissue in experimental trichinellosis.
Author's e-mail : kboczon@amp.edu.pl
Małgorzata Kotula-Balak1, Leszek Bablok2, Stanisław Frącki2, Anna Jankowska3 and Barbara Bilińska1
1Laboratory of Endocrinology and Tissue Culture, Institute of Zoology, Jagiellonian University, Cracow, 2Department of Andrology, Institute of Obstetrics and Gynecology, Medical Academy, Warsaw, 3Department of Radiobiology and Cell Biology, University of Medical Sciences, Poznan, Poland
Abstract: Klinefelter’s syndrome (47, XXY) is the most common chromosome aneuploidy in men and is usually characterized by underdeveloped testes and sterility. The aim of the present study was to detect cellular distribution of androgen receptors (AR) and aromatase in testes of patient with KS. The tissue sections were processed for morphological and immunohistochemical staining. Additionally, levels of FSH, LH, PRL, estradiol, and testosterone were measured in the plasma. Morphological analysis revealed a complete absence of spermatogenesis. No germ cells were present in seminiferous tubules. In some tubules, nests of apparently degenerating Sertoli cells were found. In the interstitium, Leydig cell hyperplasia was observed. Using immunohistochemistry, nuclear AR staining was detected in Sertoli cells and peritubular cells, whereas in Leydig cells the staining was exclusively cytoplasmic. The immunostaining of aromatase was detected in the cytoplasm of Sertoli cells and Leydig cells. Increased levels of gonadotropins and decreased level of testosterone concomitantly with the cytoplasmic localization of AR in Leydig cells might contribute to the impaired testicular function in patient with KS.
Author's e-mail: bbili@zuk.iz.uj.edu.pl
Ewa Czeczuga-Semeniuk1, Sławomir Wołczyński1, Milena Dąbrowska2, Janusz Dzięcioł3 and Tomasz Anchim1
1Department of Gynaecological Endocrinology, 2Department of Hematological Diagnostics and 3Department of Anatomy, Medical University, Białystok, Poland.
Abstract: Doxorubicin (Adriamycin) is the most active drug in the treatment of breast cancer. The aim of this study was to investigate the interaction of doxorubicin and retinoids in the inhibition of proliferation of hormone sensitive (ER+) human breast cancer cell line MCF-7 and to find out whether this combination can result in the enhancement of its therapeutic effect. As a comparison we also used estradiol and tamoxifen. We also made an attempt to elucidate the effect of these compounds on the stimulation of the apoptotic pathway in breast cancer cells. Cell proliferation in a 24-hour culture was assessed by [3H] thymidine incorporation into cancer cells and by immunocytochemical analysis of cellular cycle-related PCNA and Ki-67 antigens expression, after the incubation of the cell culture with 10, 20 and 50 nM doxorubicin (DOX), 2 nM estradiol (E2), 10 μM tamoxifen (TAM) and 1 nM, 0.01, 0.1, 1 and 10 μM of all-trans retinoid acid (ATRA). The assessment of cell viability and analysis of apoptotic and necrotic cells were performed after the 72-hour incubation of the culture with the examined substances and following apoptosis induction using acridine orange and ethidine bromide. Of the doxorubicin concentrations used in the study, 20 nM inhibited thymidine incorporation to 84.83±10.00% (control=100%). In the same culture conditions, 2 nM E2 stimulated cancer cells to 157.09±8.84%. Concentrations of 10 μM TAM and 10 μM ATRA inhibited the proliferation to 63.16±7.85% and 52.19±3.21%, respectively. A statistically significant reduction of these values was observed when 20 nM DOX was added to medium with E2 - 39.24±7.6%, TAM - 48.34±2.05% and ATRA - 21.98±1.69%, respectively; the percentage of PCNA- and Ki-67-positive cells was also reduced. Despite high antiproliferative efficacy of 20 nM DOX and 10 μM ATRA combination, the percentage of apoptotic cells was only 25±0.81%, being similar to that obtained in the culture with 20 nM DOX. The concentrations of 10, 20 and 50 nM DOX that were used to inhibit the proliferation of MCF-7 cell line were not particulary effective. The inhibitory effect was obtained when 20 nM of DOX and E2, TAM or ATRA were used simultaneously. The use of E2 caused a two-fold decrease in the percentage of proliferating cells. It was also shown that the effectiveness of DOX in combination with ATRA is significantly higher than that of DOX combined with TAM, which might suggest a valuable approach to the treatment of breast cancer.
Author's e-mail: czeczuga@wp.pl
E. Iżycka-Świeszewska1, W. Kloc2, K. Plata-Nazar3, J. Stefanowicz4, E. Drożyńska4 A.Woźniak5, D.Gąsecki6 and M. Dębiec-Rychter7
1Department of Pathology, Medical University, Gdansk,
2Neurosurgery Clinic, Polish Hospitals, Gdansk,
3Department of Pediatrics, Pediatric Gastroenterology and Oncology, Medical University, Gdansk,
4Department of Pediatrics, Pediatric Hematology, Oncology and Endocrinology, Medical University, Gdansk,
5Department of Biology and Genetics, Medical University, Gdansk,
6Department of Neurology, Medical University, Gdansk, Poland,
7Centre for Human Genetics, Catholic University, Leuven, Belgium
Abstract: Four cases of primitive neuroectodermal tumors (PNETs) with unusual localization (three intraspinal extramedullary and one pontocerebellar) are reviewed. Histologically, they were small round blue cell tumors with diverse patterns. Immunohistochemically, all tumors were positive for at least two neuronal markers, two cases were Mic-2 positive and one showed glial differentiation. The paraffin-embedded tumor specimens were examined by interphase FISH using dual-color probes specific for EWS, HER-2 and BCR loci. Molecular cytogenetic study revealed the presence of EWS rearrangement in two cases and the presence of i(17q) in one tumor. Three tumors exhibited 22 disomy and one was 22 polyploid. Extraparenchymal PNETs within craniospinal axis are heterogeneous from the clinical, histological, immunohistochemical and molecular point of view. These PNETs can be of a central or peripheral type. Multidisciplinary approach is of a basic importance in differential diagnosis of such cases.
Author's e-mail: eczis@wp.pl
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The morphological analysis of vasculature in thyroid tumours: immunoexpression of CD34 antigen
Marta Rzeszutko1, Wojciech Rzeszutko1 and Piotr Dzięgiel2
1Department of Pathological Anatomy and 2Department of Histology and Embryology, University Medical School, Wrocław, Poland
Abstract: Angiogenesis represents an important process manifested in tumour growth and in development of metastases. Using immunohistochemistry, the authors evaluated number of vessels in various nodular lesions of the thyroid (54 cases). Expression of CD34 antigen and microvessel density were evaluated in sections of archival paraffin blocks originating from the Department of Pathological Anatomy, University Medical School and the Lower Silesia Centre of Oncology in Wrocław, Poland. Microvessel density was assessed in ten different fields per section in “hot spots”. Expression of CD34 was quantified using computerised image analysis and, then, mean micrvessel count (MVC) and microvessel area (MVA) were calculated. In thyroid tissue with benign lesions, the MVC (31.7) was higher than in neoplastic lesions (22.3), although no differences in MVA were observed. This observation points to differences in the size of newly formed vessels in individual nodular lesions of the thyroid.
Author's e-mail: piotr@hist.am.wroc.pl
Grzegorz Mazur1, Tomasz Wróbel1, Piotr Dzięgiel2, Michał Jeleń3, Kazimierz Kuliczkowski1 and Maciej Zabel2
1Department of Haematology, Blood Neoplasms and Bone Marrow Transplantation, 2Department of Histology and Embryology, and 3Department of Pathology, Wrocław Medical University, Wrocław, Poland
Abstract: Angiogenesis is important in development, maintenance and progression of haematological malignancies. Some clinical observations have indicated that in non-Hodgkin's lymphoma (nHL) tumour microvessel density (MVD) may correlate with tumour staging and outcome. The aim of the study was to examine relationship between MVD as a parameter of tumour angiogenesis measured by expression of CD34 and the grade of nHL histological malignancy as determined by REAL classification. 40 lymph node samples of patients with newly diagnosed nHL (17 women, 23 men; aged 48-70 yrs, median age 64 yrs; stage III and IV) and treated at the Department of Haematology, Wrocław Medical University in 1999-2002 were fixed in 10% buffered formalin and embedded in paraffin. In all the studied cases, sections were incubated with antibodies against CD34. The slides were stained with hematoxylin and eosin and evaluated histopathologically. Patients were divided into two groups according to histological malignancy: indolent nHL (19 patients) and aggressive nHL (21 patients). Mean MVD measured by expression of CD34 in aggressive and indolent nHL groups amounted to 19.45±11.24 vessels/0.375 mm2 and 21.7±12.4 vessels/0.375 mm2, respectively. Statistical analysis of microvessel staining demonstrated no correlation between tumour MVD and grade of histological malignancy in lymph nodes of nHL patients. Nevertheless, angiogenesis observed in nHL provides rationale for use of angiogenesis inhibitors in lymphoma therapy.
Author's e-mail: grzegmaz@hemat.am.wroc.pl
BackMarek Pawlikowski1, Anna Gruszka1, Maciej Radek3 and Jolanta Kunert-Radek2
1Department of Experimental Endocrinology and Hormone Diagnostics, 2Department of Clinical Endocrinology, Institute of Endocrinology, and 3Department of Neurosurgery and Surgery of Peripheral Nerves, Medical University, Łódź, Poland
Abstract: Forty one pituitary adenomas excised surgically were immunostained to reveal pituitary hormones and chromogranin A (CgA). In 23 patients, plasma CgA concentration was determined before surgery by ELISA method. The CgA immunopositivity was found in 70.7% of investigated tumors. It was observed in all tumors of gonadotropinoma type and in the majority of null cell adenomas. Elevated (>18 U/L) plasma CgA concentration was observed in approx. a half of the examined patients, being more frequent in gonadotropinomas and null cell adenomas. It may have some, although limited, diagnostic value in these types of pituitary tumors.
Author's e-mail: pawlikowski.m@wp.pl
Grażyna Hoser1, Danuta Wasilewska1 and Joanna Domagała-Kulawik2
1Department of Clinical Cytology, Medical Center of Postgraduate Education, 2Department of Pneumonology and Allergology, Medical University, Warsaw, Poland
Abstract: In recent years many data indicate that lymphocytes from cancer patients undergo increased apoptosis. The objective of this study was to evaluate the expression of Fas receptor on lymphocytes obtained from patients with lung cancer. Eighteen patients with non-small cell lung cancer and 18 healthy volunteers were investigated. Expression of Fas (CD95) on CD4+ and CD8+ blood lymphocytes was evaluated by flow cytometry. The proportion of blood Fas+ lymphocytes was significantly higher in lung cancer patients when compared with healthy individuals and in smokers when compared with nonsmokers.
Author's e-mail: graho@cmkp.edu.pl
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