[Record linkage of a large clinical practice patient cohort with the Cancer Registry Schleswig-Holstein]. / Abgleich einer großen Patientinnen-Kohorte aus der klinischen Praxis mit dem Krebsregister Schleswig-Holstein.

INTRODUCTION: A precondition for the evaluation of outcomes in cohort studies and screening programmes is the availability of follow-up data. In Germany, established cancer registries provide such data for incident primary cancer diseases and mortality. To utilise these cancer registry data a person's identifying code has to be correctly linked to study or programme records, a procedure which, up to date, has been only rarely used in Germany. Exemplarily, the feasibility and validity of record linkage of a cohort of 173 050 patients from the Quality-assured Mamma Diagnostic programme (QuaMaDi) to the cancer registry Schleswig-Holstein was assessed by the accuracy of the classified outcome. METHODS: Name, date of birth and address of the QuaMaDi cohort members were coded in the confidential administration center of the registry. These codes were passed by the codes of 129 455 female cancer registry records. Datasets were synchronised for each match, so that QuaMaDi participants could be identified in the registry file. In a next step epidemiological registry records were linked to the QuaMaDi study records. The accuracy of classifying outcome was assessed by agreement measures, i. e., Cohen's kappa. In cases of disagreement, a questionnaire has been sent to QuaMaDi patients' gynaecologists to validate the final diagnosis. RESULTS: Synchronisation of both cohorts resulted in 18 689 one to one matches with any kind of malignant tumour, therein 8 449 breast cancers (ICD-10 C50, D05). Absolute agreement between files according to diagnosed or suspected breast cancer was 97.6% with a kappa value of 0.79. When suspicious BIRADS 4 cases from QuaMaDi were excluded, agreement and kappa rose to 99.5% and 0.948, respectively. After correction of the final diagnosis according to the physician's responses, agreement measures slightly improved in both groups of ascertained diagnosis including and excluding the suspected cases. CONCLUSION: Within QuaMaDi the diagnosed breast cancer cases were predominantly notified in the cancer registry. Discordant matches (false negatives and false positives) may have resulted due to various causes, thereof a very low percentage of record linkages from different persons. In conclusion, synchronisation of study cohort files to registry files using pseudonymous personal data is feasible and valid. The generated combined datasets can be used for comparative analysis of several objectives. One of them will be the evaluation of screening programmes in the near future.

Hepatic apo E expression is required for remnant lipoprotein clearance in the absence of the low density lipoprotein receptor.

According to the secretion-capture model of remnant lipoprotein clearance, apo E secreted by hepatocytes into the space of Disse serves to enrich the remnants with a ligand for receptor-mediated lipoprotein endocytosis. Current evidence supports a two-receptor model of lipoprotein removal, in which apo E-containing remnants bind either the low density lipoprotein receptor (LDLR) or the LDLR-related protein (LRP). Recently, we demonstrated that reconstitution of apo E(-/-) mice with apo E(+/+) marrow results in normalization of plasma lipoprotein levels, indicating that hepatic expression of apo E is not required for remnant clearance and calling into question the relevance of the secretion-capture mechanism. To dissect the relative contributions of LDLR and LRP to the cellular catabolism of remnant lipoproteins by the hepatocyte, bone marrow transplantation (BMT) was used to reconstitute macrophage expression of apo E in mice that were null for expression of both apo E and the LDLR. Reconstitution of macrophage apo E in apo E(-/-)/LDLR(-/-) mice had no effect on serum lipid and lipoprotein concentrations, although it produced plasma apo E levels up to 16-fold higher than in C57BL/6 controls. Immunocytochemistry of hepatic sections revealed abundant staining for apo E in the space of Disse, but no evidence of receptor-mediated endocytosis of remnant lipoproteins. Transient expression of human LDLR in the livers of apo E(+/+)--> apo E(-/-)/LDLR(-/-) mice by adenoviral gene transfer resulted in normalization of serum lipid levels and in the clearance of apo E-containing lipoproteins from the space of Disse. We conclude that whereas the LDLR efficiently clears remnant lipoproteins irrespective of the site of origin of apo E, endocytosis by the chylomicron remnant receptor (LRP) is absolutely dependent on hepatic expression of apo E. These data demonstrate in vivo the physiologic relevance of the apo E secretion-capture mechanism in the liver.

Macrophage lipoprotein lipase promotes foam cell formation and atherosclerosis in vivo.

Expression of lipoprotein lipase (LPL) by the macrophage has been proposed to promote foam cell formation and atherosclerosis, primarily on the basis of in vitro studies. LPL-deficient mice might provide a model for testing the role of LPL secretion by the macrophage in an in vivo system. Unfortunately, homozygous deficiency of LPL in the mouse is lethal shortly after birth. Because the fetal liver is the major site of hematopoiesis in the developing fetus, transplantation of C57BL/6 mice with LPL-/- fetal liver cells (FLCs) was used to investigate the physiologic role of macrophage LPL expression in vivo. Thirty-four female C57BL/6 mice were lethally irradiated and reconstituted with FLCs from day 14 LPL+/+, LPL+/-, and LPL-/- donors. No significant differences were detected in plasma levels of post-heparin LPL activity or in serum cholesterol or triglyceride levels between the 3 groups on either a chow diet or an atherogenic diet. After 19 weeks on the atherogenic diet, aortae were collected for quantitative analysis of the extent of aortic atherosclerosis. LPL expression was detected by immunocytochemistry and in situ hybridization in macrophages of aortic atherosclerotic lesions of LPL+/+-->C57BL/6 and LPL+/--->C57BL/6 mice, but not in LPL-/--->C57BL/6 mice, whereas myocardial cells expressed LPL in all groups. The mean aortic lesion area was reduced by 55% in LPL-/--->C57BL/6 mice compared with LPL+/+-->C57BL/6 mice and by 45% compared with LPL+/--->C57BL/6 mice, respectively. These data demonstrate in vivo that LPL expression by macrophages in the artery wall promotes foam cell formation and atherosclerosis. off

Palladium 1D nanoscale aggregates on a graphite surface using CTAB hemicylindrical micelle templates.

Nanoscale palladium clusters in the form of parallel strips have been formed on the surface of graphite with the help of a surface micellar template of cetyltrimethylammonium bromide using a chemical deposition method. The repeat period of the palladium strips deposited at 25 °C is 65 nm, with a width of 40 nm and height of 2 nm. The elemental composition of the metal clusters was confirmed using X-ray fluorescence analysis and TEM-EDX. The fact that the strips are composed of metallic palladium was also confirmed by testing the membrane electrode assembly with the strips in a commercial fuel cell. Using the obtained micellar template, the radius of the curvature of the AFM probe tip was estimated with the help of a unique method. The radius is equal to 10 nm and matches the value provided by the manufacturer.

Liposome uptake by cultured macrophages mediated by modified low-density lipoproteins.

We have investigated effects of native low-density lipoproteins (LDL) and malondialdehyde-treated LDL on the interaction of 5(6)-carboxyfluorescein-labeled liposomes bearing antibodies to LDL with cultured J774 macrophages. It was found that an addition of modified LDL to the incubation medium resulted in 15-20-fold increase of carboxyfluorescein binding to cells, whereas native LDL did not produce such effect. The increase of carboxyfluorescein binding to macrophages in the presence of modified LDL was not due to an enhanced leakage of the label from liposomes. The modified-LDL-mediated binding of carboxyfluorescein to cells was reduced to 20-30% of the initial level in the presence of cell-respiration inhibitors (NaF and antimycin A). Fluorescent microscopy data also indicate the modified-LDL-induced incorporation of liposome contents into cells. The results obtained in this study make it possible to assume that in the presence of malondialdehyde-treated LDL, liposomes with antibodies to LDL may be incorporated into macrophages via the receptor-mediated pathway for modified LDL.

Isoform-specific effects of apolipoprotein E on atherogenesis: gene transduction studies in mice.

BACKGROUND: We recently used a bone marrow-based gene therapy approach to show that small amounts of retrovirus-derived human apolipoprotein E3 (apoE3) produced by macrophages are protective against early atherosclerosis in apoE-deficient mice. METHODS AND RESULTS: In the present study, we evaluated whether the effect produced by macrophage-derived apoE3 is related to its ability to bind cellular membranes. To this end, we used apoE2 and apoEcys142, dysfunctional human variants with reduced binding to the LDL receptor or to heparan sulfate proteoglycans, respectively. ApoE-deficient mice, 5 weeks of age, received transplants of apoE(-/-) bone marrow cells transduced with either parental retrovirus or apoE3, apoE2, or apoEcys142 retroviral vectors. Human apoE was detected by ELISA in the serum of apoE3, apoE2, and apoEcys142 mice as early as 4 weeks after bone marrow transplantation, and at 8 weeks, plasma apoE levels were 55.5+/-20.3, 50.5+/-8.7, and 15.3+/-7.3 microgram/dL, respectively. In all groups, cholesterol levels increased with age but were not affected by apoE expression. As previously demonstrated, the lesion area in male apoE3 mice (3808+/-2224 micrometer(2)/section) was 40% smaller than that in control mice (6503+/-3475 micrometer(2)/section). In apoE2 mice, however, the lesion area was similar to that of controls (5991+/-2771 micrometer(2)/section), and apoEcys142 mice showed an unexpected and significant increase in lesion size (10 320+/-6128 micrometer(2)/section). Thus, transplantation with marrow transfected with receptor binding-defective apoE variants did not replicate the antiatherogenic effect of apoE3. CONCLUSIONS: These data provide in vivo evidence suggesting that macrophage-derived apoE delays development of atherosclerosis through a receptor-dependent pathway.

Reduced atherosclerotic lesions in mice deficient for total or macrophage-specific expression of scavenger receptor-A.

The absence of the scavenger receptor A (SR-A)-I/II has produced variable effects on atherosclerosis in different murine models. Therefore, we examined whether SR-AI/II deficiency affected atherogenesis in C57BL/6 mice, an inbred strain known to be susceptible to diet-induced atherosclerotic lesion formation, and whether the deletion of macrophage SR-AI/II expression would modulate lesion growth in C57BL/6 mice and LDL receptor (LDLR)(-/-) mice. SR-AI/II-deficient (SR-AI/II(-/-)) female and male mice on the C57BL/6 background were challenged with a butterfat diet for 30 weeks. No differences were detected in plasma lipids between SR-AI/II(-/-) and SR-AI/II(+/+) mice, whereas both female and male SR-AI/II(-/-) mice had a tremendous reduction (81% to 86%) in lesion area of the proximal aorta compared with SR-AI/II(+/+) mice. Next, to analyze the effect of macrophage-specific SR-AI/II deficiency in atherogenesis, female C57BL/6 mice were lethally irradiated, transplanted with SR-AI/II(-/-) or SR-AI/II(+/+) fetal liver cells, and challenged with the butterfat diet for 16 weeks. In a separate experiment, male LDLR(-/-) mice were reconstituted with SR-AI/II(-/-) or SR-AI/II(+/+) fetal liver cells and challenged with a Western diet for 10 weeks. No significant differences in plasma lipids and lipoprotein profiles were noted between the control and experimental groups in either experiment. SR-AI/II(-/-)-->C57BL/6 mice, however, had a 60% reduction in lesion area of the proximal aorta compared with SR-AI/II(+/+)-->C57BL/6 mice. A similar level of reduction (60%) in lesion area was noted in the proximal aorta and the entire aorta en face of SR-AI/II(-/-)-->LDLR(-/-) mice compared with SR-AI/II(+/+)-->LDLR(-/-) mice. These results demonstrate in vivo that SR-AI/II expression has no impact on plasma lipid levels and that macrophage SR-AI/II contributes significantly to atherosclerotic lesion formation.

Phenotype related changes of intimal smooth muscle cells from human aorta in primary culture.

To study the functional characteristics of smooth muscle cell (SMC) phenotypes, we have investigated myosin expression, cell proliferation, collagen production and low-density lipoprotein (LDL) receptor activity in intimal SMCs of normal human aorta during their growth in primary culture. By staining with rabbit antibodies to smooth muscle myosin (ASMM) 3 cell types could be distinguished in culture: homogeneously stained cells, cells with discontinuous myosin fibrils and myosin-negative cells. The ratio of cell types greatly changed with culture growth: on days 5, 7 and 14 it was 82:1:17%, 70:5:25% and 10:30:60%, respectively. After 5-6 days of culture intimal SMCs began to proliferate and DNA-synthesizing nuclei were seen 1.5-4.3 times more frequently in myosin-negative cells than in cells with homogeneous myosin distribution. At that time the number of cells reacted with monoclonal antibody (MAb) to an epitope shared collagen types I and III started to increase. By double immunofluorescence staining it was shown that the cultured cells containing both ASMM and MAb markers were found 2.0-4.8 times more rarely than MAb-positive staining in myosin-negative cells. During the first 5 days in culture LDL binding and uptake were diminished in intimal cells with intercellular lipid inclusions independently of their myosin staining pattern, but their activity increased with culture growth. Thus, SMCs from human aortic intima change their phenotype on days 6 and 7 in primary culture as manifested by alteration of myosin expression, increased cell proliferation, collagen production and LDL receptor activity. Changes in myosin expression, however, are not an essential prerequisite for cell proliferation and collagen production.

Monocyte/macrophage accumulation and smooth muscle cell phenotypes in early atherosclerotic lesions of human aorta.

In a search for early atherosclerotic lesions, we have investigated grossly normal areas of human thoracic aortas taken at autopsy from 40 trauma victims aged from 3 to 40 years. Two areas of aorta were compared: lesion predisposed to atherosclerosis (LP) area localized on the dorsal aspect of the vessel along the row of intercostal branching sites, and lesion resistant (LR) area located on the ventral aspect of the vessel. Accumulation of apolipoprotein B (apo B) was found in LP aortic area of each child older than 6 years. Similar retention of apo B in LR area appeared only in aortas of teenagers. The apo B staining increased with age in both areas tested but was usually of a greater extent in LP area than in LR area. Typical smooth muscle cells (SMCs) and a few monocytes/macrophages (Mn/Mph) were revealed in the intimal layer of all aortas examined. The number of Mn/Mph dramatically increased in LP areas of individuals over 17 years. Quantitative study of double stained sections has shown a 2- to 6-fold enhanced number of Mn/Mph in LP area compared with LR aortic area of 10 men over 21 years. Focal infiltration of Mn/Mph in aortas of young adults occurred without endothelial denudation. In addition, some intimal SMCs in LP area of 12 aortas out of 29 expressed desmin and contained well-developed endoplasmic reticulum, while such cells were seldom detected in LP area of the vessels. Thus, focal accumulation of apo B with subsequent Mn/Mph infiltration and SMC phenotypic modulation in LP aortic area of young adults may be causally involved in fatty streak and atherosclerotic plaque formation.

Primary culture of endothelial cells from atherosclerotic human aorta. Part 1. Identification, morphological and ultrastructural characteristics of two endothelial cell subpopulations.

Endothelial cells (EC) were harvested by 0.1% collagenase treatment for adult human thoracic aortas obtained 1-3 h after sudden death. At least 35-70% of EC were removed from the intimal surface of aorta, 90-95% of them being viable. Plating efficiency was 70-80%. Monolayer formation was achieved at a seeding density of 5-8 X 10(2) cells/mm2. The cells were identified as endothelium by the presence of Factor VIII antigen, Weigel-Palade bodies and typically endothelial morphology at confluence. Unlike endothelial cultures derived from human umbilical veins and infant aortas, primary cultures obtained from human adult aortas contain multinuclear EC with Factor VIII antigen and Weibel-Palade bodies. The number of multinuclear EC in cultures isolated from aortas affected by atherosclerosis was 2-fold higher (P less than 0.05) than in cultures obtained from grossly normal aortas taken from donors of the same age. EC with numerous lipid inclusions revealed by oil-red-O staining were present in all the EC primary cultures derived from aortas affected by atherosclerosis. No oil-red-O-positive cells were detected among the EC cultured from infant aorta, aorta of young donors, and umbilical vein. An electron microscopic examination of EC from atherosclerotic aorta in culture and in situ failed to reveal any ultrastructural peculiarities distinguishing multinuclear EC from the mononuclear EC.

Localization of apolipoprotein E in normal and atherosclerotic human aorta.

To elucidate the role of apolipoprotein E (apo E) in atherogenesis, we have investigated the localization of apo E in normal and atherosclerotic aortas as well as in other tissues of 32 post-mortem individuals. Using double immunofluorescence it has been found that normal intima of individuals older than 20 years and some adolescents contained immunoreactive material that reacted with poly- and monoclonal antibodies to apo E. A staining pattern of apo E differed from that of apolipoprotein B, the latter being seen in normal intima of each child older than 7 years. Apo E was present extracellularly in lipid streaks and atheromatous plaques, where its staining was particularly intensive around the necrotic zone of plaques. Some macrophages in the plaques of 4 aortas exhibited apo E-positive staining, while aortic endothelial and smooth muscle cells never contained apo E. Apo E-positive staining was not found in the majority of vessel cells, it was always, however, observed in other types of cells including hepatocytes. Kupffer cells, spleen macrophages and cerebral astrocytes. Our findings indicate that only some macrophages in human aorta may be responsible for the production of apo E that can participate in reverse cholesterol transport. At the same time, apo E accumulation in the aortic wall may promote the development of atherosclerosis.

Identification of intimal subendothelial cells from human aorta in primary culture.

Subendothelial cells (SEC) were obtained from the inner intimal layer of adult human aorta by collagenase treatment. SEC were identified in primary culture either as smooth muscle cells by staining with FITC-labeled antisera against human smooth muscle myosin or as macrophages, foam cells and contaminating endothelial cells by their uptake of malondialdehyde treated low density lipoproteins labeled with fluorescent dye 3,3'-dioctadecylindocarbocyanine. Between 1 and 5 days in culture, along with smooth muscle cells (SMC, 38-82%), endothelial cells (0-9%), macrophages and foam cells (2-32%), one more type of cell was found. This cell type resembled SMC in size and shape, but was not stained by antisera to SMC myosin. By ultrastructural criteria these cells were characterized as modulated SMC for they contained prominent rough endoplastic reticulum and Golgi complex together with basement membrane and a large number of plasmalemmal vesicles. Like SMC they reacted with phalloidin and were stained by anti-vimentin but not by anti-desmin monoclonal antibodies. The proportion of such cells varied from 5 to 33% of total cell number and increased in parallel to macrophages and foam cells in vessels with well developed atherosclerotic lesions. We conclude that the applied technique may be used for identification of cultured vascular cells including modulated SMC.

The expression of skin-specific gene K51 in the epidermal layer of human skin and in basal cell carcinoma cells.

Gene K51 probe isolated previously from the rat genomic library has been used to study the expression of its human counterpart by in situ hybridization and Northern blot analysis. A polyA-containing transcript of human gene K51 of 3 kb size has been detected in embryonic skin. The gene is also expressed in the epidermis of newborn humans and adults, but not in the adjacent mesenchymal tissues. Immunostaining with keratin antisera revealed predominantly earlier stage expression of K51 than cytokeratin markers. Sebaceous and sweat glands also contain cells expressing K51 gene. K51 expression was found in the cells of eight individual basal cell carcinomas tested, with the level of expression lower than in keratinocytes from normal human epidermis. We propose that K51 gene expression could serve as a convenient marker for the study of the process of skin keratinocyte development and the changes in this process associated with skin cancers and dysplasia.

Cell death in atheromatous plaque of the carotid artery occurs through necrosis rather than apoptosis.

Thrombogenic plaques associated with acute myocardial infarction and sudden cardiac death are relatively acellular but the mechanisms responsible for cell loss are uncertain. To help elucidate this, we investigated advanced atherosclerotic lesions taken from 13 carotid arteries. The number of cells detected by the technique for identifying DNA fragmentation (occurring at apoptosis and necrosis) known as terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end-labeling (TUNEL) technique varied in arteries from 9.6% to 40.3% and was 4 times higher in plaque (26.4 +/- 2.9%) than in the adjacent media (6.0 +/- 1.3%). The majority of TUNEL+ cells (75.5 +/- 5.6%) were smooth muscle cells (SMCs) as they also reacted with antibodies to SMC alpha-actin and myosin. TUNEL+ nuclei were found in macrophages (CD68+ cells), T-cells (CD3+ cells) and neovascular endothelial cells (von Willebrand factor: vWf+ cells) as well. From 13.0% to 34.0% (24.6 +/- 1.7%) of plaque cells stained with antibody to proliferating cell nuclear antigen (PCNA) whereas the number of PNCA+ nuclei in the adjacent media was 6 times lower (4.1 +/- 0.7%). In plaques, the number of PCNA+ cells correlated with the number of TUNEL+ cells (r = .66; p < .001) but the proportion of PCNA+ SMCs was relatively low (39.1 +/- 5.2%). Electronmicroscopic examination showed the presence of cells with condensed chromatin and convoluted cell surfaces in all arteries tested and the numbers of cells exhibiting these destructive alterations varied from 15.2% to 35.5% but no cells with typical expression of apoptotic death were detected. The results obtained indicate that all vascular cell types undergo cell death, with some prevalence in locations close to the plaque core and that intimal cells die through necrosis rather than apoptosis. We speculate that the imbalance in SMC death and proliferation may lead to acellular plaque formation.

Ultrastructural identification of cells with dendritic cell appearance in atherosclerotic aorta of apolipoprotein E deficient mice.

The present electron microscopic study was undertaken to see whether cells with a dendritic cell appearance accumulate in atherosclerotic lesions of apolipoprotein E (apoE) deficient mice. Atherosclerotic aortas from 7 eight-month old apoE deficient mice were examined. In atherosclerotic plaques as well as in the underlying media and adventitia, cells with a dendritic cell appearance including the presence of a unique tubulovesicular system were detected. The tubulovesicular system was most hypertrophied in their cellular processes where the continuous cisterns sometimes formed circular structures. The cells containing the tubulovesicular system lacked lysosomes and phagolysosomes and their cytoplasm was free of lipid inclusions. The present observations suggest that dendritic cells are involved in apoE deficient mouse atherosclerosis. ApoE deficient mice might be a useful model for investigating functions of dendritic cells in atherogenesis.

[Cross-reacting antigens of the mammalian ovular zona pellucida]. / Perekrestno-reagiruiushchie antigeny blestiashchei obolochki iaitsekletok mlekopitaiushchikh.

Antigens of zona pellucida (ZP) of different mammalian species, including the man, were studied by means of various immunochemical methods. The analysis was carried out using rabbit antisera to the pig, mouse, guenon and Java macaque eggs. After immunoadsorption (by blood serum and tissue extracts) these anti-ZP-sera reacted with water-insoluble ZP components only (in immunofluorescence). The adsorbed anti-ZP-sera were specific not only to homologous ZP, but also to ZP of other mammalian species. The spectrum of antigens of the ape ZP was most closely related to that of human ZP. The pig eggs contained antigens common with the human ZP as well. The antiserum to the mouse ZP did not react with ZP of man, ape, pig, cow, rabbit and other species under study. The blood of sterile women contained the antibodies reacting (in immunofluorescence) with ZP of mouse, pig, guinea pig, sheep and man (very weakly). The data obtained suggest the complexity of antigenic mosaic of the mammalian ZP including both "cross-reacting" (common) antigens and species specific ones.

[Use of the immunoperoxidase method on paraffin sections]. / Primenenie immunoperoksidaznogo metoda na parafinovykh srezakh.

Using an indirect immunoperoxidase technique, the methods of fixation and treatment of paraffin sections were explored for subsequent identification of myosin of smooth and skeletal muscle cells, actin, filamin, and immunoglobulins. Bouin's fluid and ethanol were found to be the most adequate fixators for immunomorphological analysis. Pronase treatment of paraffin sections increased the intensity of specific reactions considerably. Incubation of paraffin sections with normal goat serum and hydrogen peroxide decreased nonspecific staining of the sections.

[Regenerative reactions of myocardiocyte nuclei in ischemic heart disease]. / Regeneratornye reaktsii iader kardiomiotsitov pri ishemicheskoi bolezni serdtsa.

The ploidy of myocardiocytes nuclei in the left ventricle was determined stereometrically and microspectrophotometrically in 20 cases of ischemic heart disease (IHD) and 9 control cases. An increase in the number of nuclei and the degree of their ploidy and their correlation with the myocardium mass were established. The number of polyploid nuclei was particularly high in the periphery of postinfarction scars. These data indirectly indicate the participation of polyploidization and amitosis of myocardiocytes nuclei in the development of myocardial regenerative hypertrophy.

[Mitotic division of the nuclei of the human heart myocytes after healing of myocardial infarct]. / O mitoticheskom delenii iader miotsitov serdtsa cheloveka pri zazhivlenii infarkta.

Mitoses of nuclei of myocytes of the left ventricle of the heart observed in two elderly people who had died of extensive relapsing infarction are described. All the mitoses were pathological (delaying and dispersing of chromosomes in metaphase multipolar mitosis) and apparently they were completed by formation of multiple micronuclei.

[Effect of trimecaine on myocardial contractility and the rhythmoinotropic properties of the myocardium in hypoxia]. / Deistvie trimakaina na sokratimost i ritmoinotropnye svoistva miokarda pri gipoksii.

In the papillary muscles from 14 adult rats and myocardial specimens from 14 newborn ones, trimecaine in doses of 1.10(-6) and 5.10(-6) g/ml produced a dose-dependent negative inotropic effect, reversed the rhythmoinotropic correlation into a negative one and minimized the efficiency of postextrasystolic potentiation. Trimecaine in the former dose reduced hypoxic damage of myocardial specimens in the both groups as reflected by a smaller amplitude of hypoxic contracture and better contractility recovery following a hour hypoxia. It is suggested that reduction in hypoxic contracture and damage to the myocardium occur in the presence of trimecaine from limited Ca2+ entry into myocytes via the Na-Ca turnover mechanism.