[Treatment of patients with burns in surgical clinics of Leningrad hospitals in 1920-1930s]. / Lechenie bol'nykh s ozhogami v khirurgicheskikh klinikakh meditsinskikh institutov Leningrada v 1920­1930-kh godakh.

OBJECTIVE: To reconstruct the events of early period of Soviet school of combustiology (1920-1930s) via analysis and systematization of clinical and experimental studies devoted to burns and performed in the Leningrad medical institutes in 1920-1930s. MATERIAL AND METHODS: We analyzed various reports by employees of the Leningrad medical institutes devoted to practice and theory of burn treatment within the above-mentioned historical period. RESULTS: Analysis of Soviet and foreign reports for 1920-1930s made it possible to systematize data on the treatment of burns in the Leningrad medical institutes for the period from the mid-1920s to the beginning of the Great Patriotic War. We presented experimental data on local and general processes following burn injuries. CONCLUSION: We discovered and introduced into scientific circulation some reports of Leningrad scientists on clinical and theoretical aspects of burn injuries which fell out of the field of view of modern researchers for various reasons. These data emphasize diverse work carried out by the staff of surgical and theoretical departments regarding the treatment of burn injuries.

[Peculiarities of ferrorkinetis in adolescent girls with iron-deficiency anemia].

On the basis of follow up of 462 adolescent girls of 14-19 years old, in 110 of them iron-deficiency anemia was revealed. With the help of general blood analysis and biochemistry studies there was detected a disturbance of iron metabolism, declaring itself in decreasing iron serum indices, serum ferritin saturation factor and increase of general iron combined ability levels, latent combined ability and levels of serum erythropoietin.

[Changes in the type and frequency of chromosome associations in human peripheral blood lymphocytes after treatment with colcemid]. / Izmenenie kharaktera i chastoty khromosomnykh assotsiatsii v limfotsitakh perifericheskoi krovi cheloveka pri deistvii koltsemida.

Changes in the character and frequency of chromosome associations in metaphase cells of human lymphocyte cultures during colcemid treatment were studied. As the time of colcemid treatment was extended from 0 to 24 h, the number of both cells with chromosome associations and of chromosomes involved in this process was seen to increase. After a 1 h incubation of cells with colcemid the number of nucleolar-organizing chromosome associations increased considerably. In 3 h, non-nucleolar-organizing chromosomes were also involved in associations: among these most often were noticed chromosomes 1, 2 and 3, and less often chromosome 11. Following 24 h of colcemid influence, the number of telomeric associations of chromosomes increased, with most frequent involvement of chromosomes 1, 4, 6 and 16. Results of this investigation allow to propose that the step-by-step involvement of metaphase chromosomes in associations, as the time of colcemid treatment increases, may reflect the remoteness of the respective chromosomes from the nucleolus. Thus, in human blood lymphocytes chromosomes 1, 2, 3 and 11 are very likely localized near the nucleolus, or some regions of these chromosomes may contact with the nucleolus.

Karyotypic evolution of cells in culture: a new concept.

The Chapter summarizes peculiarities of karyotypic variability during establishment and long-term cultivation of permanent cell lines. A new concept on pathways of karyotypic evolution of cells in culture is put forward. A detailed description is presented of the author's original approach of cytogenetic analysis of cell lines provided for a principally new characteristic of the cell line: its generalized reconstructed karyotype (GRK). Its use as a criterion to evaluate authenticity, purity, and stability of cell lines is discussed. Based on analysis of the GRK, two stages of karyotype evolution of cell lines are revealed: establishment and stabilization, different in karyotypic variability of the cell population and in peculiarities of clone selection. Comparison of peculiarities of karyotypic variability of leukemic and tumor cells both in vitro and in vivo was made, and general regularities of their karyotypic evolution have been established, such as nonrandom changes in the number and structure of chromosomes and deletion of one of the sex chromosomes, as well as regularities characteristic only of cells in culture in most human and animal cell lines (at least 85%) of disomy on all autosomes. The rest of the cell lines, 15%, are characterized by either partial or total monosomies on certain autosomes during long-term cultivation. Three main compensatory mechanisms of maintaining viability of cell lines that have lost genetic material are discussed: polyploidization of the initial cell clone, amplification of oncogenes (predominantly of mys family), and extracopying of whole autosomes or of their fragments.

[The patterns of the karyotypic evolution of cells in culture]. / Zakonomernosti kariotipicheskoi évoliutsii kletok v kul'ture.

Numerous personal and literary data on the karyotypic variation of cell lines during their establishing and long-term culturing have been reviewed. A new notion about karyotypic evolutionary pathways of cells in culture is presented. A detailed original approach to cytogenetic study of permanent cell lines is given, which allows, via karyotype reconstruction, to obtain finally a new karyotypic characteristics-the generalized reconstructed karyotype (GRK). Application of the cytogenetic approach as a criterion for the control of authenticity, purity and stability of cell lines is discussed. The cell line analysis by means of GRK elicited that the cell line evolution in vitro passes through two stages: a stage of establishment, and a stage of stabilization, both differing in karyotypic variability of cell populations and clonal selection in culture. The data indicate that it is important in experiments to utilize cell lines being in the stage of stabilization and to characterize chromosomally the cell line at the same passage when it is used. Above all, a comparison of karyotypic variations of tumor and leukemic cells in vitro and in vivo has revealed their common karyotypic evolution regularities (a nonrandom character of numerical and structural chromosome changes and the loss of one of the sex chromosomes) and the karyotypic evolutionary regularities characteristic solely of cells in culture. The main of these being a balanced chromosome set in the cell population as a whole and obligatory retention of diploidy in all chromosomes of the normal set by the majority of human and animal cell lines. It has been revealed that no less than two homologs of each autosome are present in cells of at least 85% of examined lines. Other cell lines (at least 15%) of a generally neurogenic origin are shown to be notable by keeping partial or complete monosomies on autosomes throughout the long-term culturing. Peculiarities of the karyotypic evolution of the latter are regarded in detail in addition to the data on the expression of oncogenes and other growth-associated genes in their cells. It is suggested that there are three main compensatory mechanisms through which cell lines with autosomal monosomies may maintain the vitality in culture: polyploidization of initial cell clones, oncogene amplification, mainly of the myc-family oncogenes, and fragmentary or complete extracopying of several autosomes. In summary, perspective of cell line cytogenetics as a field of biology of the cell in culture is discussed.

[Cholera in Makhachkala during the 1994 epidemic]. / Kholera v Makhachkale v period épidemii 1994 g.

During the period of epidemic in the Daghestan 51 patients and 27 Vibrio carriers were detected in Makhachkala. A considerable proportion (30.7%) of cholera cases caused by infection imported from regions, unfavorable with respect to cholera, in the presence of pronounced migration of the population was registered. The role of different transmission routes in cases of cholera was as follows: day contacts were responsible for 43.3%, the alimentary route for 28.4% and the water route for 14.9% of cases. The epidemic situation was characterized by a mild and prolonged type of the epidemic process. Mass diseases were prevented by a complex of cholera control measures, among them the emphasis was made on various limitations and prophylactic measures aimed at the rupture of the transmission routes of V. cholerae.

Silver nucleolar organizer region assessment in tumor-cells from breast-cancer patients treated by extremely-low-frequency magnetic-field.

This study assessed the value of silver staining of nucleolar organizer regions (AgNORs) in tumor cells as a potential technique for the estimation of the action of ELF magnetic field on breast cancer. The light and electron microscopy was used for analysis of tissue specimens from 4 breast cancer patients after ELF magnetotherapy and 18 untreated persons with malignant and benign breast lesions. All patients had surgical removal of neoplasms. The mean number of AgNORs and the distribution of cells according to the AgNOR number in tissue samples from patients after successful ELF magnetotherapy were closer to those of persons with benign tumors, as compared to malignant ones. The effect of ELF magnetic field at the electron microscopic level is discussed.

[Localization of the CD4 receptor gene in the chromosomes of clone cells of the monocytoid line U-937 characterized by different sensitivities to HIV]. / Lokalizatsiia gena retseptora CD4 v khromosomakh kletok klonov monotsitoidnoi linii U-937, kharakterizuiushchikhsia razlichnoi chuvstvitel'nost'iu k VICh.

Karyotypes of two clones of U-937 line, with high and low sensitivity to HIV-1, were studied. The CD4-receptor gene-cellular receptor of HIV-1 was mapped. CD4-receptor gene was located according to in situ hybridization method, in locus 12 p11-p12, both in cells of high-sensitive clone U-937/16, and in cells of low-sensitive clone U-937/4. It is determined that in both the clones chromosomes 12 are presented in two copies and are not affected by rearrangements. That allows to conclude that the sensitivity of cells U-937 to HIV-1 does not depend on the dose of this gene, or on its transference in chromosomes.

[The interrelation between the morphological characteristics of the nucleolar apparatus in cultured human embryonic fibroblasts and the proliferative activity of the cells]. / Vzaimosviaz' mezhdu morfologicheskimi kharakteristikami iadryshkovogo apparata kul'tiviruemykh émbrional'nykh fibroblastov cheloveka i proliferativnoi aktivnost'iu kletok.

A relationship between the number of AgNOR, nucleolar area, the number of nucleoli per nuclei, the number of nucleoli touching nucleolar membrane and the proliferating activity of human embryonal fibroblast culture was investigated. Cell proliferation was arrested by serum deprivation. In 10 days, initiation of proliferation was performed by cessation of serum deprivation. The strongest correlation (r = 0.96) was found between the index of labeling (IL) and the number of nucleoli touching nucleolar membrane. A strong correlation was also observed between AgNOR count, nucleolar area and IL. The number of silver grains in the interphase nucleus decreased slowly and increased quickly, with quick decrease and increase of cell proliferation occurring, respectively. No correlation was found between the number of nucleoli and IL.

[The cytogenetic characteristics of human B-lymphoblastoid cell lines]. / Tsitogeneticheskaia kharakteristika B-limfoblastoidnykh kletochnykh linii cheloveka.

Permanent suspension B-cell lines HSL-1, HSL-2, HSL-3, HSL-4, HSL-5, HSL-8, HSL-9 have been obtained spontaneously from lymphocytes and cells of hematopoietic tissues of patients with various forms of leukemia and tumours. All cell lines contained Epstein-Barr virus (EBV). At early stages of cultivation (6-39th passages) normal karyotypes were established for all the tested lines which testified their origin from normal B-lymphocytes. In the process of cultivation of HSL-1 cell line, which (up to the 63rd passage) contained primarily the normal karyotypes, a substitution for tetraploid ones (92, XX, YY) by the 132nd passage was established. In line HSL-4, a pathological clone of cells with structural chromosome rearrangements (46, XY, -2, -12, der (12) t (2; 12) (p12, p13); del2 (p12), was revealed at the 26th passage. During a prolonged cultivation (up to the 130th passage) this abnormal clone forced out completely the clone with normal karyotype. Substitution of normal clone cells by cells with pathological karyotypes may point to their selective advantage.

[The cytogenetic characteristics of B-lymphoid lines from Papio hamadryas baboons and Macaca arctoides brown macaques]. / Tsitogeneticheskaia kharakteristika B-limfoidnykh linii pavianov gamadrilov Papio hamadryas i makakov burykh Macaca arctoides.

A cytogenetic analysis of 9 permanent suspension cell lines was performed. Cell lines were obtained from Papio hamadryas and Macaca arctoides cells by different methods. Five cell lines originated from non-malignant parental cells. Lines SPG-5 and SPG-6 were analysed at early stages of cultivation, with normal karyotypes being identified. In cell lines PL-11, MAL-1 and MAL-4 pathological clones with structural and numerical chromosomal rearrangements appeared only following a long-term cultivation. This phenomenon may be due presumably to the influence of cultivation on karyotype variability. In cell line LUG-4 chromosome marker 17p+ was found at early stages of cultivation. In sublines E9-1, E1-1 and E5-1, established from line LUG-4 by cloning, 17p+ chromosomal marker was found persisting. The presence of the marker 17p+ in all the cells at early passages testifies that the cell material from Papio with lymphoma, which was used for this line, might have the same chromosomal damage in its initial karyotype.

Increased frequency of acrocentric chromosome association during colcemid treatment.

The effect of Colcemid on satellite association frequencies was investigated in human cell lines U-937 GTB, SC-N-MC, HL-60, and Raji by silver staining method. Cell cultures were exposed to Colcemid at a concentration of 0.02 mg/ml for 15-60 minutes. As exposure time to Colcemid was increased, both the frequency of cells with satellite associations and the number of chromosomes involved in satellite associations increased significantly. Furthermore, the mean number of silver-stained chromosomes decreased with longer exposure time. Without Colcemid, we were able to obtain an adequate number of good-quality metaphase spreads. The data obtained may be of help in furthering research concerning satellite associations and in obtaining better quality chromosome preparations.

[The variability of the pattern of human G-band chromosomes in the late prophase-early metaphase]. / Izmenchivost' risunka G-okrashennykh khromosom cheloveka v pozdnei profaze--rannei metafaze.

Original modification of the harvest procedure, hypotonic treatment and slide-making techniques were used to obtain prometaphase spreads of good quality fitted to G-banding or to FISH. Human blood cultures were synchronized with a methotrexate block during synthesis, and following this with a thymidine release. Cells were then proceeded into prometaphase and early metaphase and were quickly harvested without exposure to colcemid. Following incubation in a mixture of 0.56% KCl and 1% sodium citrate (1:1) for 20-23 min at room temperature, cells were fixed in 4 changes of fixative made of 3 parts of absolute methanol and 1 part of glacial acetic acid. Three or four 100-105 microliters volume of a suspension of fixed cells were blew with a strength through pasteur pipette tip from a distance 5-6 cm onto chilled superclean slides wetted by water. These slides were then quickly flame dried. Prior to staining, prometaphase bearing spreads were held in thermostat overnight at 37 degrees C. Prometaphases were G-banded with the aid of a modified technique elaborated in the Department of Clinical Genetics, Lund University Hospital (Sweden). The slides were incubated for 1-4 min at room temperature in 0.01 versene solution, containing 0.011% w/v trypsin (Difco), 0.4 mg/ml D-glucose, 0.17 mg/ml KCl, 1.7 mg/ml NaCl. Following a 15 sec incubation in a solution containing 1 mg/ml D-glucose, 0.4 mg/ml KCl, 8 mg/ml NaCl, G-banding was completed by staining the slides with 2% Giemsa (Merck) in pH 6.8 phosphate buffer for 4-5 min.(ABSTRACT TRUNCATED AT 250 WORDS)

[The dependence of the quantity of silver granules detectable in interphase nuclei after the silver nitrate staining of the chromosomal nucleolus-organizer regions on the conditions for the hypotonic treatment of the cells and the effect on this index of cell treatment by a pharmacological agent causing dissociation of the nucleolar components]. / Zavisimost' kolichestva granul serebra, vyiavliaemykh v interfaznykh iadrakh posle okrashivaniia iadryshkoobrazuiushchikh raionov khromosom nitratom serebra, ot uslovii gipotonicheskoi obrabotki kletok i vliianie na étot pokazatel' obrabotki kletok farmakologicheskim agentom, vyzyvaiushchim dissotsiatsiiu komponentov iadryshka.

We investigated effects of hypotonic treatment of cultured HL-60 cells and human fibroblasts, and of DRB on the quantity of AgNORs revealed following AgNO3 staining. The average number of grains increases after KCl hypotonic treatment. The lower the concentration of KCl in the solution, the higher the number of silver grains revealed in the nucleoli due to their dissociation into substructures. The use of adenosine analogue (DRB) before the hypotonic treatment of cells facilitates the following count of silver deposits. A maximum increase in the mean number of silver grains was observed after a 1.5 h incubation.

[The structure and function of the chromosomal nucleolus organizer regions: the molecular, cytological and clinical aspects]. / Struktura i funktsiia iadryshkoobrazuiushchikh raionov khromosom: molekuliarnye, tsitologicheskie i klinicheskie aspekty.

Numerous data concerning nucleolar organizer region (NOR) structure and function in normal and pathological human cells are analysed. The contents of argentophilic nucleolar proteins vary in the cells tested. The silver-stained test is closely connected with cellular proliferative potentials and differentiation status of the cells. This test also depends on the level of tumor cell malignancy and seemed to be very useful for megakaryocyte and cardiomyocyte studies. In summary, the NOR silver-staining approach is of great value for both fundamental cytological investigations and wide medical practice.

[The use of DNA hybridization in situ for identifying chromosomal rearrangements in the karyotyping of cell lines]. / Ispol'zovanie gibridizatsii DNK in situ dlia identifikatsii khromosomnykh perestroek pri kariotipirovanii kletochnykh linii.

A complex karyological analysis of three human cell lines (PLC-PRF-5, MT-4 and U937) was carried out that involved traditional methods of G-, C-banding and silver staining, in addition to the in situ hybridization technique using 4 alpha-satellite DNA probes: DNA specific for centromeric regions of chromosome 11, 6, 13, and 21, and 14 and 22. The application of this additional method allowed to identify, prove or detalize the structure of 13 markers in PLC-PRF-5 cells, 1 marker in MT-4 cells, and 3 markers in U937 cells. The results show that the in situ hybridization method would be successful in cell line karyotyping for a more objective identification of some markers difficult for analysis.

AgNOR staining in benign hyperplasia and carcinoma of the prostate.

We have modified existing techniques for silver staining of nucleolar organizer regions of intact interphase cells by hypotonic swelling and by formic acid treatment to reduce background staining. This allowed the microscopic identification and counting of individual AgNORs in the nucleoli. The method was used on nine adenomatous prostatic samples (including one of normal prostate tissue outside a localized tumor) and on seven prostatic adenocarcinomas. In general, the adenomatous samples displayed fewer AgNORs (mean 13 dots/cell) than did the carcinomas (mean 24 dots/cell). Although no cells with very high AgNOR counts were found in specimens from nonmalignant tumors, two of the adenomatous prostates did have AgNOR profiles that to a large extent overlapped with those of carcinomas. A highly differentiated carcinoma (of which only very small amounts were present in the sample) had low AgNOR counts. The three moderately differentiated carcinomas had more silver-positive material than the nonmalignant prostates but less than the three poorly differentiated carcinomas. The latter tumors also had a substantial proportion of cells with greater than 60 AgNOR counts, something that was never seen in carcinomas with higher differentiation. The data indicate that analysis of silver staining-positive material in intact interphase cells may help distinguish between benign and malignant prostatic tumors and between highly malignant and low malignant carcinomas.