M.Z. Ratajczak, M. Kucia, M. Majka, R. Reca, J. Ratajczak: Heterogeneous populations of bone marrow stem cells - are we spotting on the same cells from the different angles? pp. 139-146
B. Turyna, A. Jurek, K. Gotfryd, A. Siaśkiewicz, P. Kubit, A. Klein: Peritonitis-induced antitumor activity of peritoneal macrophages from uremic patients pp. 147-153
B. Lewczuk, M. Nowicki, M. Prusik, B. Przybylska-Gornowicz: Diurnal rhythms of pinealocyte ultrastructure, pineal serotonin content and plasma melatonin level in the domestic pig pp. 155-163
A. Augustynowicz, J. Dzięcioł, M. Barwijuk-Machała, J. Dadan, Z. Puchalski, S. Sulkowski: Assessment of proliferative activity of thyroid Hürthle cell tumors using PCNA, Ki-67 and AgNOR methods pp. 165-168
A. Dąbrowski, A. Filip, W. Zgodziński, M. Dąbrowska, D. Polańska, M. Wójcik, K. Zinkiewicz, G. Wallner: Assessment of prognostic significance of cytoplasmic survivin expression in advanced oesophageal cancer pp. 169-172
W. Kruszewski, R. Kowara, R. Rzepko, C. Warężak, J. Zieliński, G. Gryglewski, A. Kopacz, T. Jastrzębski, T. Pawełczyk: K-RAS point mutation, and amplification of C-MYC and C-ERBB2 in colon adenocarcinoma pp. 173-179
B. Zawadowska, J. Majerczak, D. Semik, J. Karasinski, L. Kolodziejski, W. M. Kilarski, K. Duda, J. A. Zoladz: Characteristics of myosin profile in human vastus lateralis muscle in relation to training background pp. 181-190
N. Keklikoglu: The localization of Fos B, a member of transcription factor AP-1 family, in rat odontoblasts and pulpal undifferentiated ectomesenchymal cells pp. 191-193
J. Woźniak: Two methods for the quantitative analysis of surface antigen expression in acute myeloid leukemia (AML) pp. 195-199
ABSTRACTS
Heterogeneous populations of bone marrow stem cells - are we spotting on the same cells from the different angles?
Mariusz Z. Ratajczak1,2, Magda Kucia1, Marcin Majka1, Ryan Reca2 and Janina Ratajczak2
1European Stem Cell Therapeutic Excellence Center, Medical College, Jagiellonian University, Cracow, Poland
2Stem Cell Biology Program at James Graham Brown Cancer Center and Department of Medicine, University of Louisville, Louisville, USA
Abstract: Accumulated evidence suggests that in addition to hematopoietic stem cells (HSC), bone marrow (BM) also harbors endothelial stem cells (ESC), mesenchymal stem cells (MSC), multipotential adult progenitor cells (MAPC), pluripotent stem cells (PCS) as well as tissue committed stem cells (TCSC) recently identified by us. In this review we discuss the similarities and differences between these cell populations. Furthermore, we will present the hypothesis that all of these versatile BM derived stem cells are in fact different subpopulations of TCSC. These cells accumulate in bone marrow during ontogenesis and being a mobile population of cells are released from BM into peripheral blood after tissue injury to regenerate damaged organs. Furthermore, since BM is a “hideout” for TCSC, their presence in preparations of bone marrow derived mononuclear cells should be considered before experimental evidence is interpreted simply as trans-differentiation or plasticity of HSC. Finally, our observation that the number of TCSC accumulate in the bone marrow of young animals and their numbers decrease during senescence provides a new insight into aging and may explain why the regeneration processes becomes less effective in older individuals.
Author's e-mail: mzrata01@louisville.edu
Bohdan Turyna1, Aleksandra Jurek1, Kamil Gotfryd1, Agnieszka Siaśkiewicz1, Piotr Kubit2 and Andrzej Klein1
1Department of General Biochemistry, Faculty of Biotechnology, Jagiellonian University, and 2Department of Nephrology and Dialysis, Rydygier Hospital, Cracow, Poland
Abstract: The macrophages belong to the effector cells of both nonspecific and specific immune response. These cells generally express little cytotoxicity unless activated. The present work was intended to determine if peritoneal macrophages collected from patients on Continuous Ambulatory Peritoneal Dialysis (CAPD) during episodes of peritonitis were active against human tumor cell lines without further in vitro stimulation. We also compared macrophage antitumor potential with effectiveness of drugs used in cancer therapy (taxol and suramin). Conditioned medium (CM) of macrophages collected during inflammation-free periods did not exhibit cytostatic and cytotoxic activity against both tumor (A549 and HTB44) and non-transformed (BEAS-2B and CRL2190) cells. Exposure of tumor cells to CM of macrophages harvested during peritonitis resulted in significant suppression of proliferation, impairment of viability and induction of apoptosis, in contrast to non-transformed cells, which remained unaffected. The efficacy of CM of inflammatory macrophages as an antitumor agent appeared to be comparable to cytostatic and cytotoxic potency of taxol and suramin or, in the case of HTB44 cells, even higher. The results obtained suggest that activated human macrophages might represent a useful tool for cancer immunotherapy.
Author's e-mail: bohdan@mol.uj.edu.pl
Bogdan Lewczuk, Marcin Nowicki, Magdalena Prusik and Barbara Przybylska-Gornowicz
Division of Histology and Embryology, Department of Functional Morphology,
Faculty of Veterinary Medicine, University of Warmia and Mazury, Olsztyn, Poland
Abstract: The study was conducted to investigate diurnal changes in pinealocyte ultrastructure, pineal serotonin content and plasma melatonin concentration in the domestic pig. The immature pigs (n=24) were kept under a cycle of 12 h light : 12 h dark, with a photophase between 0800 and 2000. During the photophase the animals were exposed to direct sunlight. After four weeks the gilts were slaughtered at 0900, 1400, 2100 and 0200. The pineals were removed and divided into two parts – one for quantitative ultrastructural study (by a point count method) and one for serotonin assay. Simultaneously, blood samples were taken for melatonin assay. The relative volume of mitochondria in pinealocyte perikarya was significantly higher at 1400 than at 0200 and 0900 as well as at 2100 than at 0200. The relative volume of Golgi apparatus was higher at 0900 and 1400 than at 0200. The relative volume of dense bodies of the MBB-1 type in pinealocyte perikarya was significantly lower at 1400 and 2100 than at 0900. In contrast, the relative volume of MBB-2 was higher at 1400 than at 0900 and 0200. The numerical density of DCV in perikarya was significantly higher at 0200 than at 1400. No significant differences were found in rough endoplasmic reticulum, lysosomes and multivesicular bodies. The pineal serotonin content showed a prominent rhythm with the maximum at 1400. The plasma melatonin concentration was significantly higher at 0200 than at 0900, 1400 and 2100. The obtained results demonstrate that both pinealocyte ultrastructure and pineal biochemistry in the pig undergo significant changes in the course of the diurnal rhythm.
Author's e-mail: lewczukb@uwm.edu.pl
Albert Augustynowicz1, Janusz Dzięcioł2, Małgorzata Barwijuk-Machała3, Jacek Dadan4, Zbigniew Puchalski4 and Stanisław Sulkowski3
Department of Medical Pathomorphology1, Department of Anatomy2, Department of General Pathomorphology3, and 1st Clinic of Surgery4, Medical University, Białystok; Poland
Abstract: We have undertaken an attempt to compare the application efficacy of the proliferative activity markers in differential diagnosis of thyroid Hürthle cell tumors (HCT) using the PCNA and Ki-67 labeling and AgNOR visualisation techniques. The present work is a retrospective analysis of 78 Hürthle cell tumors: 20 Hürthle cell carcinomas (HCC), 32 Hürthle cell adenomas (HCA) and 26 hyperplastic nodules with Hürthle cell metaplasia (HCM). Five µm sections were stained according to AgNOR technique and labeled with antibodies against PCNA and Ki-67. AgNOR dot count in the nucleus and proliferative index (PI – percentage of cells expressing PCNA and Ki-67) in randomly chosen nuclei (100 in case of AgNOR and over 1000 in case of PI) were evaluated in each slide. The mean values of AgNOR dot count, PI-PCNA and PI-Ki-67 in HCC, HCA and HCM were respectively: 5.1, 61.3 and 54.9; 3.4, 42.4 and 38.6 and 2.5, 39.3 and 34.3. Statistically significant difference was found in all the proliferative activity markers between malignant and benign tumors: HCC:HCA (p<0.01) and HCC:HCM (p<0.001). There was no statistically significant difference between HCA and HCM.
Author's e-mail: albertaugustynowicz@poczta.onet.pl
A. Dąbrowski1, A. Filip2, W. Zgodziński1, M. Dąbrowska2, D. Polańska2, M. Wójcik2, K. Zinkiewicz1 and G. Wallner1
12nd Department of General Surgery and 2Department of Genetics, Skubiszewski Medical University, Lublin, Poland
Abstract: Survivin is a member of the family of proteins, which inhibit apoptosis (inhibitor of apoptosis proteins – IAP). Expression of survivin was found in colorectal cancer, neuroblastoma, bladder cancer, non-small cell lung cancer, and breast cancer. There is some recent data indicating the correlation of poor prognosis and worse response to chemotherapy in patients with oesophageal squamous cell carcinoma (OSCC) expressing survivin. The aim of the present study was to assess survivin expression in cancerous tissue of patients with advanced OSCC and to test the potential correlation between survivin expression and clinicopathological data. Forty two patients (mean age 58.36±8.97 yrs), who were oesophagectomised due to squamous cell carcinoma of the thoracic oesophagus between 1998 and 2000, were retrospectively analysed. Cytoplasmic survivin expression, examined immunohistochemically, was found in 35 (83.33%) cases. No statistically significant correlation between survivin expression in the tumour and patients' gender, TNM stage, or vascular involvement was noted. The mean survival of patients with cytoplasmic survivin expression (17.8±15.51 months) was not statistically different to those with negative survivin staining (16±6.28 months) as assessed by Mantel-Cox test (p=0.49). Univariate regression analysis revealed UICC staging as the only predictor of survival in the analysed group (p<0.05). These results indicate that the cytoplasmic survivin expression does not seem to be the prognostic factor in advanced cases of OSCC.
Author's e-mail: wzgodz@wp.pl
Wiesław Kruszewski1, Renata Kowara2, Robert Rzepko3, Cezary Warężak1, Jacek Zieliński1, Grzegorz Gryglewski1, Andrzej Kopacz1, Tomasz Jastrzębski1, and Tadeusz Pawełczyk2
1Department of Surgical Oncology, 2Department of Molecular Biology and 3Department of Pathology, Medical University, Gdańsk, Poland
Abstract: The routine multidisciplinary management of colon cancer is based mainly on tumor staging, histology, grading and vascular invasion. In this approach, important individual information derived from molecular characteristics of the tumor may be missed, especially since significant heterogeneity of molecular aberrations in cancer cells has been observed, and recognition of every of relationships between them may be of value. K-RAS, C-MYC and C-ERBB2 are protooncogenes taking part in carcinogenesis and tumor progression in the colon. They influence cell proliferation, differentiation and survival. K-RAS point mutation, as well as amplification of C-MYC and C-ERBB2 were searched in 84 primary colon adenocarcinomas resected with curative intent. Multiplex polymerase-chain reaction and restriction fragment length polymorphism were performed to assess codon 12 K-RAS point mutation. Amplification of C-MYC and C-ERBB2 genes was evaluated by densitometry after agarose gel separation of the respective multiplex PCR products. No relation was found among mutated and/or amplified genes, and between searched molecular aberrations and pathoclinical features. In multivariate analysis, nodal status appeared to be the only independent prognostic indicator. In colon adenocarcinoma, codon 12 K-RAS point mutation and amplification of C-MYC and C-ERBB2 seem to occur independently in the process of tumor progression. Amplification of C-ERBB2 tends to associate with more advanced stage of disease. Concomitant occurrence of codon 12 K-RAS mutation, C-MYC and C-ERBB2 amplification was of no prognostic value in respect to survival.
Author's e-mail: wjkrusz@amg.gda.pl
B. Zawadowska1, J. Majerczak2, D. Semik1, J. Karasinski1, L. Kolodziejski3, W. M. Kilarski1, K. Duda3, and J. A. Zoladz2
1Department of Cytology and Histology, Institute of Zoology, Jagiellonian University, 2Department of Muscle Physiology, AWF-Krakow, 3Cancer Institute, Krakow, Poland.
Abstract: Twenty-four male volunteers (mean ± SD: age 25.4±5.8 years, height 178.6±5.5 cm, body mass 72.1±7.7 kg) of different training background were investigated and classified into three groups according to their physical activity and sport discipline: untrained students (group A), national and sub-national level endurance athletes (group B, 7.8±2.9 years of specialised training) and sprint-power athletes (group C, 12.8±8.7 years of specialised training). Muscle biopsies of vastus lateralis were analysed histochemically for mATPase and SDH activities, immunohistochemically for fast and slow myosin, and electrophoretically followed by Western immunoblotting for myosin heavy chain (MyHC) composition. Significant differences (P<0.05) regarding composition of muscle fibre types and myosin heavy chains were found only between groups A (41.7±1.6% of MyHCI, 40.8±4.0% of MyHCIIA and 17.5±4.0% of MyHCIIX) and B (64.3±0.8% of MyHCI, 34.0±1.4% of MyHCIIA and 1.7±1.4% of MyHCIIX) and groups A and C (59.6±1.6% of MyHCI, 37.2±1.3% of MyHCIIA and 3.2±1.3% of MyHCIIX). Unexpectedly, endurance athletes (group B) such as long-distance runners, cyclists and cross country skiers, did not differ from the athletes representing short term, high power output sports (group C) such as ice hockey, karate, ski-jumping, volleyball, soccer and modern dance. Furthermore, the relative amount of the fastest MyHCIIX isoform in vastus lateralis muscle was significantly lower in the athletes from group C than in students (group A). We conclude that the myosin profile in the athletes belonging to group C was unfavourable for their sport disciplines. This could be the reason why those athletes did not reach international level despite of several years of training.
Author's e-mail: zaw@zuk.iz.uj.edu.pl
Nurullah Keklikoglu
Department of Histology and Embryology, Faculty of Dentistry, Istanbul University, Istanbul, Turkey
Abstract: It has been proposed that cellular proliferation and differentiation are accomplished by AP-1 components but different components can be responsible for different functions. In this study, our aim was to compare the localisation of Fos B, which is a component of AP-1, in postmitotic differentiated and undifferentiated cells via Fos B immunoreactivity. For this purpose, maxillary incisor teeth from 10 Wistar rats were obtained and Fos-B was investigated immunohistochemically in formalin-fixed, paraffin-embedded tooth sections containing odontoblasts, which are postmitotic differentiated cells, and pulpal undifferentiated ectomesenchymal cells. No significant differences in percentage of Fos B-positive cells were observed between the two cell types (p > 0.05). These findings suggest that Fos B, a component of AP-1 family, seems to have a negligible effect on differentiation and proliferation in odontoblasts and pulpal undifferentiated ectomesenchymal cells.
Author's e-mail: nkeklik@istanbul.edu.tr
Jolanta Woźniak
Department of Physiopathology, Institute of Haematology and Blood Transfusion, Warsaw, Poland
Abstract: The expression of lineage molecules (CD13 and CD33), c-Kit receptor (CD117), CD34, HLA-DR and adhesion molecule CD49d was assessed in acute myeloid leukemia (AML) blast cells from 32 cases, using direct and indirect quantitative cytometric analysis. High correlation (r=0.8) was found between antigen expression intensity values calculated by direct analysis method (ABC) and by indirect analysis method (RFI). Moreover, the differences in expression intensity of CD13, CD117 and CD34 antigens were found between leukemic and normal myeloblasts. This may be helpful in identification of leukemic cells in the diagnostics of minimal residual disease after treatment in AML patients.
Author's e-mail: facsiht@ihit.waw.pl