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20th Anniversary of Association Doctors of Hope (ADH) is an opportunity for presentation of the conclusions resulting from the realization of our tasks, especially those related to the Polish medical communities in the Borderlands. Maintaining the contacts with the Polish minorities in Lithuania, Latria, Byelorussia, Ukraine, Romania, Moldavia and Georgia are the priority tasks of ADH. Those communities differ in the extent of their needs and the relations with the local authorities in the context of the aid provided by our Association and the cooperation with us. The programme of our medical missions in the Borderlands included: a) rendering immediate aid by supplying medicines, medical materials; b) physical examination of groups of Poles (including the youths); c) organizing joint medical and pedagogical conferences concerning heath problems and healthy lifestyle. The article refers to the need for coordination of the cooperation among Polish organizations, the organizations in the Borderlands and consulates, in order to facilitate studying of the Poles in Poland, as well as to arrange for specialization residencies in Poland for doctor of Polish descent. Perfecting of the professional qualifications is a vital factor in strengthening the respect for the Polish communities, which, as minorities, are continuously expose to breaking the law by the local authorities.

The aim of this study was to determine and to evaluate silica induced lung cell reactivity--if any--in bronchoalveolar space, before clinical changes develop. Bronchoalveolar lavage (BAL) was carried out in 15 nonsmoking individuals with chronic professional silica exposure, free of lung signs and symptoms. Controls were healthy nonsmokers. Routine BAL cytology (HE, MGG) was completed by mast cell staining (toluidine blue). BAL lymphocyte subsets were phenotyped by direct two- and three-color immunofluorescence (applied DAKO A/S monoclonal antibodies: anti-CD3, CD4, CD8, CD11b, CD14, CD15, CD16 + 56, CD19, CD25, CD45, HLA-DR). Parallel staining was performed in peripheral blood. In individuals with chronic silica exposure we found: significant increase in alveolar macrophage (362 +/- 45 vs 160 +/- 33 x 10(3) cells/ml, p < 0.05), lymphocyte (61 +/- 9 vs 24 +/- 5 x 10(3) cells/ml, p < 0.05) and BAL total cell (415 +/- 76 vs 187 +/- 34 x 10(3) cells/ml, p < 0.05) numbers; significant increase in mast cell (0.4 +/- 0.1 vs 0.2 +/- 0.1, p < 0.05), NK cell (7.0 +/- 1.8 vs 3.6 +/- 1.0, p < 0.05) and Th early activated lymphocyte percent (CD4 + CD25+ calculated as percentage of CD4+ cells: 15.1 +/- 1.5 vs 7.8 +/- 1.6, p < 0.01). All results were presented as median +/- SEM. Bronchoalveolar space of people with chronic silica exposure usually shows pathological reaction (especially macrophagic alveolitis), although they are free of manifested pulmonary disease. Th early activated lymphocytes, NK cells and mast cells seem to play important role in the early interstitial lung tissue reaction to silica.

In this paper we considered the value of correctly performed bronchoalveolar lavage (BAL) in diagnosis of interstitial lung diseases and in assessment of the activity of pathological process. We indicated the conditions of exact interpretation of the results of BAL cytoimmunological examination, i.e. fine standard handling of BAL material, including its collection, elaboration and choice of lavage site, as well as regarding external (e.g. cigarette smoking) and internal additional factors. We described the influence of BAL fluid recovery and of the method of staining on obtained results. We emphasized routinely performed BAL to be the valuable diagnostic and research tool in pulmonology however, the method may have limited usefulness and unnecessarily burden the patient, if technical guidelines are not observed.

Here, using our original technique for measuring of thrombogenesis on the surface of rabbit aorta endothelium superfused with whole blood, we demonstrate that the thrombogenic property of endothelium is potentiated in the course of 3 months cadmium intoxication. The loss of endothelial thromboresistance is accompanied by suppressed generation of endogenous prostacyclin, leukopenia, increased platelet aggregability and by the presence of quasi-atherosclerotic, focal proliferative, glassy-protein lesions formed in aortic endothelium. We hypothesize that the final loss of vessel thromboresistance with all sequence of events that accompany cadmium intoxication, may result from the cadmium-induced inhibition of the generation of endothelial prostacyclin. However, the exact mechanism by which cadmium intoxication may affect the generation of prostacyclin and, then, functioning of blood platelets and vascular endothelium requires further investigations.

The analysis of mast cells in bronchoalveolar lavage (BAL) is rarely performed in routine diagnostic protocol. The BAL mast cells are rather difficult to be identified due to their paucity and technical problems with fixation and staining methods. The present study was carried out in order to characterize quantitatively and morphologically of mast cells obtained in bronchoalveolar lavage from 38 patients with several lung disorders. The percentage of mast cells ranged from: 0.0-0.6% (aspirin intolerant asthmatics), 0.1-1.8% (aspirin tolerant asthmatics), 0.9-1.8% (sarcoidosis), 1.5-2.0% (fibrosis), 0.04-0.08% (normals). There were statistically significant differences in the mast cells count between tested disease groups. Lung mast cells exhibited heterogeneity of size, shape, intensity of staining and degree of degranulation. Bronchoalveolar lavage is a very interesting and suitable model for the study of human lung mast cells. These cells should be analysed because of their clinical importance in pathogenesis of several lung disorders. We recommend, Carnoy's solution as a fixative and 0.5% solution of Toluidine blue (pH-0.5 with prolongation of staining time to 5-7 days at 4 degrees C) as a dye for routine diagnostic method of BAL mast cells identification.

Objective: To analyze various argyrophilic nucleolar organizer regions (AgNOR) parameters in proliferative lesions of the thyroid gland and to determine to what extent these parameters are influenced by the type of lesion or malignancy., Study Design: AgNORs were studied by light microscopy with the help of a semiautomated image analysis system in the normal thyroid gland (n = 6), nodular hyperplasia (n = 12), thyroid hyperfunction (n = 6); follicular adenoma (n = 21), oxyphilic adenoma (n = 10), follicular carcinoma (n = 10), oxyphilic carcinoma (n = 9), papillary adenocarcinoma (n = 14), and giant (n = 6) and small cell anaplastic carcinoma (n = 4). The investigated parameters included the number and area of AgNOR dots, the number of separate AgNOR locations, the ratio of the AgNOR dot area to the nuclear area and the area of single AgNOR dots., Results: The number and area of AgNOR dots were low in all the lesions, with the exception of giant cell anaplastic carcinoma. The number of separate AgNOR locations was the lowest in oxyphilic cell tumors and medium in papillary adenocarcinoma, while in all other lesions and normal thyroid it had higher values. The ratio of AgNOR area to the nuclear area and the area of single dots were similar in proliferative lesions and normal thyroid. In all proliferative lesions and in normal thyroid, there was a significant positive correlation (P < or = .0001) between the area or number of AgNOR dots on the one hand and nuclear area on the other., Conclusion: The results indicate that in thyroid proliferative lesions, AgNOR values, particularly the number of separate AgNOR locations, correlate with the type of lesion but not with malignancy. Very typical are single clusters of dots within nuclei in oxyphilic tumors, malignant and benign, and in papillary adenocarcinomas, more or less often accompanied by single dots.

In the present study various AgNOR parameters were assessed in the computer-aided image analysis and involved the normal epithelium of colon both superficial and epithelium of crypts, adenomas and carcinomas of colon in humans. It indicated that the number and area of AgNOR dots was highest in carcinomas and decreased stepwise from carcinomas through adenomas and the epithelium of crypts to the superficial epithelium. Other parameters such as: the ratio of AgNOR dot area to nuclear area and the size of single dots were different only in the superficial epithelium. The number of AgNOR separate locations showed a tendency for higher values in neoplasms. In neoplastic lesions and normal epithelium there was a positive correlation (p < 0.0001) between AgNOR dot area and nuclear area.

The effect of compound 48/80, given intracerebroventricularly, on mast cells (MC) and histamine, serotonin and noradrenaline levels in the hypothalamus, thalamus and hippocampus as well as on corticosterone secretion was investigated in rats. A relatively high amount of mast cells was found in the thalamus, a very low in the hypothalamus and almost none in the hippocampus. Compound 48/80 (1 and 5 micrograms) induced MC degranulation, a significant increase in corticosterone secretion and diminution of the thalamic histamine level. The drug also elicited small biphasic changes in histamine level in the hypothalamus and moderately increased serotonin turnover in the brain structures studied, but did not affect noradrenaline level in those structures. These results indicate that brain MC contain a significant amount of histamine, but not serotonin. Compound 48/80 releases histamine from MC, but does not markedly influence neuronal biogenic amines in the brain structures involved in regulation of the hypothalamic-pituitary-adrenal axis.

Various AgNOR parameters were measured using the computer-aided image analysis in the normal bronchial epithelium (n = 13), in squamous cell carcinoma (n = 25) and in small-cell carcinoma (n = 11). The number and area of AgNOR dots were the highest in squamous cell carcinoma. In small cell carcinoma and in the normal epithelium the values were significantly lower. The number of AgNOR separate locations was higher in tumors in general. The size of single AgNOR dots showed differences between squamous cell carcinoma and small-cell carcinoma. Ratio of AgNOR dot area to nuclear area was similar in various pathological lesions and normal epithelium. There was a positive correlation ( < or = 0.0001) between the number and area of dots, the number of AgNOR separate locations, the area of single AgNOR dots on one hand and nuclear area on the other.

The effect of thalamic mast cells (MCs) degranulation and serotonin liberation by compound 48/80 on the hypothalamic-pituitary-adrenocortical (HPA) activity, measured indirectly through corticosterone secretion, was investigated in conscious rats. All drugs were given intracerebroventricularly (icv), the serotonin antagonists 15 min prior to compound 48/80. One hour after administration, compound 48/80 (1 and 5 micrograms) caused a significant increase in degranulated MC number in the thalamus, from control value of 20% up to 58%, and a considerable rise in the serum corticosterone level, but only minor diminution of the thalamic serotonin content. Pretreatment with methysergide, a serotonin receptor antagonist, only slightly dimished the compound 48/80-induced corticosterone response, while pretreatment with cyproheptadine, an antagonist of serotonin-histamine and cholinergic-receptors, significantly decreased the compound 48/80-elicited corticosterone response. These results show for the first time that thalamic mast cells contain a very small amount of serotonin, which may play only a minor role in increasing the HPA activity by compound 48/80. These findings also suggest that other mediators liberated from mast cells by compound 48/80 are responsible for the considerable increase in the HPA activity.

The aim of this study was to establish the diagnostical value of nuclear area in proliferative lesions of colonic and bronchial epithelium. The study included 11 adenomas and 9 adenocarcinomas of colon, 11 control samples of normal colonic epithelium as well as 5 cases of bronchial squamous cell metaplasia, 25 squamous cell carcinomas, 9 small cell carcinomas and 3 adenocarcinomas of bronchi and 13 control samples of normal bronchial epithelium. All tissue samples were formalin fixed, paraffin embedded and silver stained. Morphometrical analysis was performed using optical Axiophot microscope with immersion, B/W CCD camera connected to frame grabber card, computer PC AT 386 and morphometrical software Vist and Morpho. Nuclear area was measured semiautomatically. Within each case 100 nuclei were analyzed. Statistically significant differences between carcinoma and adenoma or normal epithelium were observed in colon. In bronchi nuclear area was significantly higher in squamous cell carcinoma and adenocarcinoma if compared with other lesions including small cell carcinoma or bronchial metaplasia and normal epithelium.

The present study of AgNORs was carried out by computer-aided semiautomatic image analysis and involved epithelium, both surface and glandular, from various parts of the digestive tract in rats. It indicated that the number of AgNOR dots, their area and sometimes even the ratio of AgNOR area to nuclear area or the number of AgNOR clusters varied in epithelium from different parts of the digestive tract. The number and area of AgNORs correlated with the proliferative activity of cells in a given epithelium and they showed some typical features associated with the location. It should be noted that the area of AgNOR dots per nucleus and the area of a single AgNOR dot also positively correlated with the nuclear area.

The aim of this study was to establish the diagnostic value of nuclear area in proliferative lesions of thyroid gland. This study included 98 histological preparations of thyroid and 57 cytological preparations (touch imprints). Morphometric analysis was performed using light microscopy, B/W CCD camera connected to the frame grabber card, computer PC AT 386/3 and morphometric software Vist and Morpho. Nuclear area was measured semiautomatically. Our study indicated that the nuclear area was significantly higher in cytological preparations and that size of nuclei seems to depend on the type of neoplasm and to a lesser extent on whether a given tumor was malignant or benign.

NORs are the loops of DNA that contain the sites of ribosomal genes around which the nucleolus is formed during telophase of mitosis. In light microscopy they can be visualized by simple silver staining technique as small dark dots within the nucleus. It has been recognized that in many neoplasms, especially malignant AgNORs are more numerous and often atypical when compared with benign tumors and normal tissue. We have introduced this novel technique to the fine needle cytology of thyroid neoplasms (n = 56). We have analyzed the number, the area of AgNORs, the number of clustered AgNORs in the nucleus and the ratio of AgNOR area to the nuclear area and the area of single AgNOR by means of semiautomatic computerized image analysis. We have studied cytological samples consisting of 7 simple goiters, 7 hyperplasias, 15 follicular adenomas, 7 oxyphilic follicular adenomas, 6 follicular carcinomas, 8 oxyphilic follicular carcinomas, 6 papillary adenocarcinomas. In this study we have demonstrated that some differences in the AgNORs value are associated with the type of tumor rather than with malignancy. Location of the AgNORs seems to be very typical for some types of tumors. For example in oxyphilic neoplasms they form single clusters in the nucleus and in papillary adenocarcinomas they form at least two abundant clusters. In other proliferative lesions of the thyroid gland location of AgNORs is less typical.

Preliminary studies to evaluate lymphocyte subsets bronchoalveolar lavage (BAL) and blood were carried out in a group of patients with lung asbestosis. The assessments were made by an indirect immunofluorescent method using monoclonal antibodies as a marker. The results were recorded on a flow cytofluorometer FASC-can. As compared with a control group, the patients with lung asbestosis showed a slight decrease in the total level of T lymphocytes (CD3) in BAL, however with a clearly disturbed proportion between T-helper (CD4) and T-suppressor (CD8) lymphocytes, which led to a decreased CD4/CD8 ratio. The blood level of T lymphocytes and their subsets in these patients approximated that in the controls. Heavy smoking was also found to enhance disorders in their number and proportion. The assessment of B lymphocytes (by using polyclonal antihuman immunoglobulins (sIg/FITC) showed a significantly high level of these lymphocytes in BAL with simultaneously low levels in blood in the same patients with lung asbestosis. We did not find a clear synergic effect of cigarette smoking on the levels of B lymphocytes in the studied group.

The aim of this study was the evaluation of cytological, immunocytological and cytochemical tests efficiency in describing the pulmonary alveoli cells activity in people chronically exposed to the polluted air. The detailed analysis of bronchoalveolar lavage (BAL) obtained by rinsing of pulmonary alveoli in 36 cases of prolonged exposure to silica and asbestos dust, divided into non-smokers and smokers, was performed. The control were the patients free of any pathological pulmonary symptoms, in this number the inhabitants of Cracow. The BAL cells contribution and activity was evaluated by following methods: routine cytology (the number and percentage of cells), computer cytomorphometry (for assessment macrophage area), nitro-blue-tetrazolium (NBT) reduction test (macrophage activity) and indirect immunofluorescence with use of monoclonal antibodies (macrophage subsets) and FACS-star-PLUS flow cytometer (lymphocyte subsets). In the individuals chronically exposed to the different forms of air pollution we found the changes in number and proportions of bronchoalveolar space cells, the alterations in cell activity (especially pulmonary macrophages) and decrease in CD4/CD8 lymphocyte ratio, that could indicate the apparent disturbances in the local cell resistance. According to our observation there is a synergistic action of tobacco smoking and air pollution. Finally, we may conclude, that BAL, was a non-invasive, repeatable method, useful for monitoring pulmonary changes due to dust exposure.

The influence of brain mast cells degranulation, exerted by compound 48/80 given intracerebroventricularly, on the opioid-induced stimulation of the pituitary-adrenal axis, measured indirectly through corticosterone secretion, was investigated in conscious rats. The mu- and delta-opioid receptor agonists Leu-enkephalinamide, morphine and beta-endorphin, given intracerebroventricularly, dose-dependently increased the serum corticosterone levels. The effect of Leu-enkephalinamide was not changed by pretreatment with the histamine H1- and H2-receptor antagonists mepyramine and cimetidine. When Leu-enkephalinamide, morphine and beta-endorphin were given 3 hr after compound 48/80, the serum corticosterone levels were higher than after either one of these drugs given separately, which suggests an independent mechanism of action. In rats pretreated 24 hr earlier with compound 48/80, i.e., when brain mast cells were completely degranulated, the stimulating effect of morphine was almost abolished and the effects of Leu-enkephalinamide and beta-endorphin were considerably reduced. Since the histamine levels in the whole brain and thalamus were elevated 3 and 24 hr after administration of compound 48/80, these results suggest that factors other than histamine depletion from brain mast cells by compound 48/80 may be responsible for the dramatic impairment of the stimulating effect of morphine, Leu-enkephalinamide and beta-endorphin on the pituitary-adrenal axis.

The purpose of the present study was to analyse the differences in the number Ag-NORs dots in the selected cases of carcinomas and adenomas of the large intestine as well as in the stromal cells, surface epithelium and epithelium of crypts of the large intestine. Ag-NORs dots are consider to be argyrophilic proteins associated with so called nucleolar organizer regions (NORs). The increase in their number is regarded as a marker of metabolic and proliferative changes in the cells. In our material we found a definite statistical differences in the number of Ag-NORs dots in carcinomas, adenomas and normal mucosa of the colon. The problem of adenomas grade IV with visible signs of malignancy, atypical number of Ag-NORs is, however, controversial. This may be an indicator of the different growth dynamics of these adenomas.

Investigations were carried out on the non-specific cell-mediated response in mice with transplantable tumours: LLC in mice C57Bl and F1(C57Bl X C3H), and SAL-l tumour in Balb/c mice. The malignant cells were implanted subcutaneously for observation of changes in the non-specific cell-mediated response at the time of the local growth of the tumour, and injections into the vein or into the foot-pad were made for studying the non-specific cell-mediated response at the time of tumour dissemination. In each series of the experiments the animals received an extract of calf thymus--the preparation TFX-Polfa. The control groups had the tumour cells implanted or injected but were not given TFX. For determining the non-specific cell-mediated response the animals were immunized with picryl chloride. In both experimental models, that is with subcutaneous implantation and with injections intravenously or into the foot-pad a decrease of the non-specific cell-mediated response was observed progressing gradually with extension of the tumours. Long-term administration of TFX in doses of 0.1 mg per mouse prevented the decrease of the non-specific cell-mediated response in the first place in mice with subcutaneous implantations. In mice with malignant cells injected intravenously or into the foot-pad TFX administration had no evident effect on the value of the test. This observation suggests that TFX prevents the development of a secondary immune deficit in mice with subcutaneous transplanted tumours--that is mainly at the time of local growth of the tumour, but is of little use in the phase of tumour dissemination.

The results of the investigations on the effect of highly concentrated normobaric oxygen on rabbit lung are reported. Detailed pathomorphological analysis demonstrated that rapidly developing extensive changes of the structural elements of the capillaries and other vessels were the prevalent lesions. These changes are connected, in the light of earlier experiments carried out under identical conditions, with the observed disturbances in the metabolism of pulmonary lipids and suggest pathogenetic correlations between prostaglandins and pulmonary surfactant.

Using the tail swelling test, delayed-type hypersensitivity (DTH) to tumor and self antigens accompanying the development of syngeneic polyoma tumor in CBA mice was observed. The soluble polyoma cell-surface antigen contained both polyoma and H-2k specificities and induced weak proliferation response of spleen cells in the presence of Il-2 in vitro. Appearance of a measurable tail swelling reaction at the side of the midtail, subcutaneous injection of the eliciting antigen not earlier than at 16th hour, maximum of swelling after 24 hours and the results of histological examination proved that the swelling was caused by a typical DTH to the antigens. Maximum DTH to both antigens occurred in the mice 6 days after polyoma cell transplantation (about 3 days before the appearance of palpable tumor) and weakened as the tumor progressed. DTH activity was transferred by spleen T lymphocytes to naive recipients. Five-day restimulation of splenocytes from polyoma transplanted donors with antigen in the presence of Il-2 led to an increase of antigen-lymphocytes affinity but resulted in a decrease of DTH activity of these cells. The mechanisms of these processes are still discussed.

Among the patients directed to the Department of Endocrinology of the Institute of Internal Medicine for diagnostic examinations of the thyroid, 726 (647 women and 79 men, of age between 16 and 79 years) were qualified to surgery after scintigraphic detection of cold nodules. From this group, 474 patients had thin-needle biopsy specimens taken for cytologic examination. The results of the latter examinations have been compared with those of histologic examination performed on the tissue removed during surgery. The occurrence of a malignant tumor (in 99.1%--thyroid cancer) was found in 108 patients (22.8%) from the group having thin-needle biopsy performed. Comparison of the results of cytologic examination with those of postoperative histologic examination demonstrated low sensitivity of biopsy examination (50.0%), as opposed to high specificity (96.2%) and high accuracy (85, 3%). Analysis of the false-negative results (54/108) revealed that the main causes of errors are difficulties in differentiation of adenoma from follicular carcinoma. Widening of the indications to surgery by a group of patients with adenoma will increase sensitivity to 84.8% and will require including into the operated group of about 50% of patients with scintigraphically diagnosed cold nodules: The introduction of the new term: "urgent strumectomy", has been proposed; also a diagnostic algorithm, useful for early diagnosis of malignant tumors of the thyroid, has been discussed.

Adenomatous polyps and adenocarcinomas with variety of the precancerous stages have been induced by intraectally injection of N-methyl-N'-nitro-N-nitrosoguanidine. The aim of our experiments have been to study some quantitative and qualitative changes in mucus secretion both in the neoplasm and in transitional area surrounding tumor. The changes in composition of mucus confirmed by several histochemical methods reflect the earliest morphologically observable stage of malignant transformation in colonic mucosa.

The study was carried out on 72 rats receiving during 7 weeks rectal infusions of N-methyl-N'nitro-N-nitrosoguanidine (MNNG), a carcinogenic agent. During the follow-up of 52 weeks the changes were investigated in the quantitative composition of the aerobic and anaerobic bacterial flora of the large intestine, and the basic composition of the aerobic flora was established. It was found that MNNG modified the quantity and composition of the bacterial flora of the colon not only with a reduction in the number of bacteria in the faeces but also with a transient change in the composition and proportions of various bacterial species. Attention is called to the observation that the appearance and development of malignant tumours in the large intestine was connected with a rise in the number of micro-organisms correlated with the histological transformation of the colonic mucosa and the number of tumours in it.

In rats exposed for 12 weeks to the mixture of nitric oxides (0.34--2.81 mg/m3) and chlorine (0.61--1.50 mg/m3) the following changes were found: increased methemoglobin concentration (MetHb), increased partial pressure, increased total carbon dioxide concentration (pCO2 TCO2), increased current dicarbonate concentration (AB), and increased buffer bases (BB). In addition, asparagine transferase activity (aspAT), alanine aminotransferase (A1AT), alkaline phosphatase (AP) and hepatic isoenzyme of lactic dehydrogenase (LDH5) in serum were found to be increased. Histopathological examination revealed: inflammatory lesions and edema of pulmonary parenchyma, alveolar emphysema and edema of connective tissue of palpetra derm with mastocytes. Chronic exposure to low concentrations of nitric oxides and chlorine induces, apart from local lesions in conjunctivae, pulmonary lesions leading to respiratory acidosis compensated by metabolic alkalosis, or liberation of indicatory enzymes through impaired cells.

The studies on infiltrations consisting of reactive cells, i.e. lymphocytes, macrophages and neutrophils within and in the vicinity of neoplastic tissue were performed in order to elucidate the nature of the growth and to asses the role of these cells in human tumors. The objective methods for both qualitative and quantitative evaluation of the infiltrating cells are necessary to obtain comparable results. We report our studies on the numbers and types of reactive cells present in 3 transplantable murine tumors with different immunogenicity (SaL-1, LLC, MCA-Sa). The cytological characterization of cells was performed as following: after enzymic digestion of the tissue the cell suspension was then passed through Millipore filters and dyed. The preliminary evaluation of the percentage of lymphocytes T found in the tumor infiltrations was performed using cytotoxicity test with monoclonal antithymocytic serum (Monoclonal Anti-Mouse Thy 1, 2). We have shown that there are differences in the number and type of reactive cells in infiltrations of the three different tumor tested. With the increasing mass of tumors the percent of reactive cells decreases proportionally.