Vol. 40, No. 4, 2002
CONTENTS
S. Gonkowski: Distribution of spinal sensory neurons supplying the adrenal gland in the pig
Histone mRNA in situ hybridization in assessing proliferative activity of normal and malignant cells
Jerzy Słowiński1, Urszula Mazurek2 and Grażyna Bierzyńska-Macyszyn3
1Department of Neurosurgery and Neurotraumatology, 2Department of Molecular Biology, and 3Department of Pathology, Medical University of Silesia, Bytom, Poland
Abstract: Proliferative capacity is an important determinant of tumour biological behaviour. For research and diagnostic purposes, immunohistochemical techniques are usually applied in the assessment of tissue proliferative status. An interesting alternative for these studies is a detection of histone mRNA. As the synthesis of histones is tightly coupled with DNA replication during S-phase of the cell cycle, histone mRNA level is a specific marker of S-phase cells. Furthermore, a short-lived transcript guarantees accurate estimation of S-phase cell pool at the moment of tissue fixation. The progress in molecular biology techniques during the last decade made possible the use of in situ hybridization, especially its non-radioactive version, in routine laboratory services. This technique can be successfully applied to detection of histone mRNA in routinely processed tissues. Advantages and limitations of such approach in tumour proliferation studies are discussed.
Author's e-mail: slowian@mp.pl
Jarosław Cisowski, Adrian Zarębski and Aleksander Koj
Department of Cellular Biochemistry, Institute of Molecular Biology and Biotechnology, Jagiellonian University, Kraków, Poland
Abstract: It is known that during acute phase reaction IL-6 activates STAT3 in hepatoma cells and IL-1 downregulates this response. We found that the inhibitory properties of IL-1 on STAT signalling cascade in human hepatoma HepG2 cells are considerably decreased not only in the presence of MAP kinase inhibitors SB203580 and PD98059 but also by some antioxidants (N-acetyl cysteine and pyrrolidine dithiocarbamate) and by anti-inflammatory cytokine IL-4. It appears that cytokine crosstalk between IL-6 and IL-1 includes a direct pathway sensitive to antioxidants and MAP kinase inhibitors, whereas the indirect prolonged response depends probably on synthesis of suppressors of cytokine signalling (SOCS).
Author's e-mail: koj@mol.uj.edu.pl
Jan Sikora and Jan Żeromski
Department of Clical Immunology, University School of Medical Sciences, Poznań, Poland.
Abstract: Malignant effusions in serous cavities represent suspension of tumors and inflammatory cells and therefore present an attractive model for studying tumor-host interactions. T cell functional ability is linked to proper signal transduction via T cell receptor (TCR-CD3 complex), especially its z chain. The latter has been found to be often depressed in malignancy. Aim of this study was to assess alterations of z chain expression and its relation to apoptosis of T lymphocytes. T lymphocytes obtained from pleural effusions of 20 cancer patients and 9 patients with nonmalignant effusions serving as a control were studied. The analysis was performed using flow cytometry combining CD3, CD 4, CD8, CD16/56, TCR-z and TUNEL reaction. In the control group, T lymphocytes obtained from pure lymphocyte effusions had constantly high level of z chain while those from malignant pleural effusions had a decreased z chain expression. Lymphocytes with low z chain expression were the main subpopulation of T cells undergoing apoptosis.
Author's e-mail: jsikora@ampat.amu.edu.pl
Monika Papież1, Monika Gancarczyk2 and Barbara Bilińska2
1Department of Cytobiology and Histochemistry, Pharmaceutical Faculty, Medical College, Jagiellonian University, and 2Laboratory of Endocrinology and Tissue Culture, Institute of Zoology, Jagiellonian University, Kraków, Poland
Abstract: Among medicinal plants, extract from the hollyhock flowers is a source of antocyanides and flavonoids. The latter compounds belong, among others, to phytoestrogens (plant-derived dietary estrogens). The important role of estrogens in the testis is now well documented, and phytoestrogens, which may act as estrogen agonists or estrogen antagonists can also alter the reproductive function of the male. The aim of this study was to show whether the exposure of male rats to the aqueous hollyhock extract could affect the process of aromatization in their testes and in cultured Leydig cells. This was investigated by immunocytochemistry and radioimmunological assays. Immunoreactivities for aromatase and estrogen receptor b were weaker both in testicular sections and cultured Leydig cells after hollyhock extract administration when compared to the controls, while the intensity of immunoreaction for estrogen receptor a remained unchanged. A lower level of estradiol secreted by cultured Leydig cells from the experimental group positively correlated with a direct inhibition of aromatase activity. Additionally, a quantitative analysis of flavonoid fraction from the hollyhock extract revealed the presence of quercetin and kaempferol. It seems that a weak antiestrogenic activity of flavonoid compounds present in the hollyhock extract is mediated through aromatase and estrogen receptor b rather than by estrogen receptor a .
Author's e-mail: bbili@zuk.iz.uj.edu.pl
Sławomir Gonkowski
Department of Clinical Physiology, Faculty of Veterinary Medicine, Warmia and Mazury University, Olsztyn, Poland
Abstract: The present study was aimed at disclosing which spinal ganglia contribute to the innervation of the adrenal gland in the pig with special regard to the morphology and intraganglionic distribution of the primary sensory neurons within these ganglia. To this end, the animals were injected with a retrograde fluorescent neuronal tracer Fast Blue into the left adrenal gland during laparotomy. After a survival period of three weeks, labelled spinal sensory neurons were found in the ipsilateral dorsal root ganglia (approximately 99% of all retrogradely marked neurons). However, single adrenal gland-projecting perikarya were observed also in the contralateral dorsal root ganglia from Th3 to L3. The majority of the retrogradely labelled afferent neurons (above 65% of all Fast Blue-positive (FB+) perikarya) were located in two groups of spinal ganglia (at neuromeres Th6-7 and Th12-14, approximately 18% and 47% of neurons, respectively), forming two distinct multiganglionic centres of origin for this neural pathway. The morphometric evaluation of FB+ neurons revealed that the vast majority of them (approximately 81%) belonged to the class of small-sized perikarya (10-30 mm in diameter), while the medium-sized (diameter 30–80 mm) and large neurons comprised only up to 13% and 6.5% of adrenal gland-projecting neurons, respectively. Furthermore, the analysis of the intraganglionic distribution pattern of the retrogradely labelled cells revealed that the highest number of them was located in the medio-caudal domain of the dorsal root ganglia, irrespective of the neuromere studied. Thus, the present study has revealed sources and morphological characteristic of spinal afferent neurons supplying the porcine adrenal gland, simultaneously pointing out to the characteristic features of their inter- and intraganglionic distribution pattern.
Author's e-mail: Slawomir.Gonkowski@uwm.edu.pl
Agnieszka Bossowska
Department of Clinical Physiology, Faculty of Veterinary Medicine, University of Warmia and Mazury, in Olsztyn, Poland
Abstract: The existence of afferent neurons supplying IMG nerve cells by their collaterals has previously been demonstrated in laboratory animals, where these fibers constitute an important part of intraganglionic nerve input involved in the regulation of autonomic neurons activity. The domestic pig is thought to be a reliable animal model for medical and biomedical studies, especially those dealing with the organization of nerve pathways contributing to the neural control of the bowel and lower urinary tract functions. However, there is no data dealing with the existence and distribution of spinal afferent neurons that may be involved in the integration of various stimuli at the level of IMG, one of the most important nerve centers controlling these organs. Therefore, the present study was aimed at disclosing the distribution and morphological features of sensory neurons associated with the porcine IMG by means of retrograde tract-tracing technique. To this purpose, the right IMG was injected with the retrograde neuronal tracer Fast Blue (FB) in five immature female piglets of Large Polish White race and the distribution of FB-positive neurons in dorsal root ganglia (DRG) of interest was studied three weeks later. Retrogradely labelled perikarya were found in both left and right DRG Th15 – L5, however, with a clear lateralization in the bilateral projections pattern. The number of contralaterally located IMG-projecting perikarya distinctly decreased towards the caudal direction, constituting approximately 23%, 20%, 16%, 6.5%, 4% and 0% of all FB-positive neurons found in both left and right DRG at neuromeres Th15 – L5, respectively. The majority of FB-positive neurons (approximately 90% of all traced cells) were found in bilateral L2 and L3 DRG. The largest part of IMG-projecting sensory neurons (68.9 ± 5.0%), irrespective of their location, belonged to the class of medium-sized perikarya (diameter from 30 to 80 mm). The present study provides a detailed description of distribution and morphological characteristics of spinal afferent neurons projecting to the porcine IMG, simultaneously demonstrating typical features of their inter- and intraganglionic distribution pattern. Furthermore, the present results strongly suggest that spinal afferents contribute to the co-ordination and integration of peripheral reflexes involving sympathetic IMG neurons.
Author's e-mail: Agnieszka.Bossowska@uwm.edu.pl
Elżbieta Czykier1 , Maciej Zabel2,3 and Joanna Surdyk-Zasada2
1Department of Histology and
Embryology, Medical Academy, Białystok;
2Department of Histology and
Embryology, Medical Academy, Poznań;
3Department of Histology and
Embryology, Medical Academy, Wrocław, Poland
Abstract: The present study deals with immunohistochemical localization of PTHrP in prepubertal and pubertal testis of European bison. PTHrP immunoreactivity was observed in germinal cells in the testis of both prepubertal and pubertal animals. In calves, PTHrP was found in germinal cells, in seminiferous tubules lacking the lumen. The reaction was strong and regularly distributed within the cytoplasm. In adult animals, the reaction showed differentiation in spermatogenic cells. Some cells were strongly and diffusely stained, others exhibited weaker reaction of granular pattern. Sertoli cells and Leydig cells were PTHrP-negative in calves and adult animals.
Author's e-mail: czykier@amb.edu.pl
Justyna Szumiło1, Danuta Skomra1, Daniel Chibowski1, Andrzej Dąbrowski2, Grzegorz Wallner2 and Roman Maciejewski3
1Department of Pathomorphology, 22nd Department of General Surgery, and 3Department of Human Anatomy, Medical University, Lublin, Poland
Abstract: The study was undertaken to determine the immunoexpression of protein products of nm23 genes which are thought to be potential metastasis suppressor genes, in esophageal squamous cell carcinoma, and to analyze their relationship to selected clinicopathological features (age, sex, tumour size, depth of invasion, presence of lymph nodes and distant metastasis, pathologic tumor stage, degree of cancer differentiation and vascular/lymphatic invasion), as well as to the overall survival. Immunohistochemical staining with monoclonal antibody against nm23 using LSAB2/HRP method on formalin-fixed, paraffin-embedded sections obtained from 32 tumors was performed. Eight tumors (25%) showed positive nm23 immunoexpression. There were no statistically significant differences between nm23-positive and nm23-negative groups with respect to all clinicopathological features studied. The positive nm23 status was related to a worse prognosis but this was not significant. The results may suggest that nm23-status is not associated with metastatic ability and prognosis in esophageal squamous cell carcinoma, but such thesis requires further study on a larger population.
Author's e-mail: jszumilo@wp.pl
Elżbieta Pawlicka1, Lech Romanowicz1, Edward Bańkowski1, Lech Chyczewski2 and Stefan Jaworski3
Departments of 1Medical Biochemistry, 2Molecular Biology and 3Gynaecology, Medical Academy, Białystok, Poland
Abstract: Edema, proteinuria, hypertension (EPH-gestosis), most commonly termed as pre-eclampsia, is the most common pregnancy-associated pathological syndrome. It is accompanied by a thorough remodelling of extracellular matrix in the umbilical cord tissues. It is commonly known that the presence of serum in culture medium strongly stimulates many functions of cells cultured in vitro. It was decided to check how the pre-eclamptic serum affects the fibroblast division in culture. Ki-67 is a protein present in proliferating cells and can be detected during all phases of the cell cycle (G1, S, G2/M) but not in resting (G0) cells. PCNA (proliferating cell nuclear antigen) is an intranuclear polypeptide whose synthesis rate is at its maximum during the S-phase of the cell cycle. The expression of Ki-67 and PCNA was measured by immunocytochemical methods and biosynthesis of DNA was evaluated by [14C]-thymidine incorporation. The activity of pre-eclamptic umbilical cord serum (UC-serum) was found to be distinctly lower in comparison to control one. The expression of Ki and PCNA in fibroblast cultures treated with pre-eclamptic serum was also distinctly lower. Also the incorporation of [14C]-thymidine to DNA was lower than in the cultures treated with control UC-serum. It may by concluded that pre-eclampsia reduces the mitogenic activity of the umbilical cord serum.
Author's e-mail: edward@amb.edu.pl
Stanisław J. Chrapusta1 and Michał Walski2,3
Laboratories of 1Experimental Pharmacology and 2Ultrastructure of the Nervous System, Polish Academy of Sciences Medical Research Center; 3Department of General Biology and Parasitology, Medical University, Warsaw, Poland
Abstract: A random finding of intracranial vascular Chlamydia sp. infection in Wistar rats and its associated damage of the component tissues of the basilar arterial wall are described as seen with transmission electron microscopy. The pathological changes observed showed no apparent specificity except for "striding" of the endothelial cells on the elastic lamina, the phenomenon observed in all five rats with basilar arterial Chlamydia infection, and in only two out of 13 rats with no proof of the infection. No atherosclerotic changes were found in the infected artery in rats sacrificed 8 or 26 weeks after the presumed infection.
Author's e-mail: chrapuss@medres.cmdik.pan.pl
