Prognostic significance of S-phase fraction detected by antithymidine antibodies in epidermoid cervix carcinomas.

PURPOSE: To assess the predictive value of pretreatment proliferative activity of epidermoid cervix carcinoma cells with respect to short- and long-term results of radiotherapy. METHODS AND MATERIALS: The proliferative activity of 25 epidermoid cervix carcinomas was evaluated as the immunofluorescent labeling index (LI) by rabbit antithymidine antibodies reacting specifically with single-stranded DNA of replication forks in S-phase cells. The short-term clinical outcome was estimated at 3-6 months after treatment by visual and palpatory examination. Three-year follow-up data were obtained through hospital charts and correspondence with referring physicians for only 19 patients. RESULTS: There was no statistically significant association between LI and such conventional prognostic factors as clinical stage. The LI value of cervix carcinomas was significantly associated with complete regression at 3-6 months after radiotherapy and 3-year disease-free survival. Complete regression at 3-6 months was observed in 87.5% patients with fast proliferating tumors (LI > 7.0%), and only in 41.2% patients with slowly proliferating tumors (p = 0.03). Probability of 3-year disease-free survival was 85.7% in patients with fast proliferating tumors and 50.0% in those with slowly proliferating tumors (p = 0.05). CONCLUSIONS: The immunofluorescent LI of epidermoid cervix carcinoma is able to provide prognostic information on short-term tumor response to radiotherapy and disease-free survival.

A monoclonal antibody to DNA modified with osmium tetroxide/2,2'-bipyridine.

A murine monoclonal antibody (IgG) has been generated that binds to DNA modified with osmium tetroxide in the presence of 2,2'-bipyridine and does not interact with unmodified DNA. Reactivity of the antibody was tested by gel retardation assay, ELISA, dot-binding assay and immunoblotting. The results obtained suggest that the antibody does not cross-react with modified or unmodified RNA or proteins. The high specificity of the binding reaction is due to the specific recognition of modified deoxythymidine residue by the monoclonal antibody. A possible way of using the antibody produced is discussed.

Antibodies to natural double-stranded RNA and to synthetic polyribonucleotides in the sera of patients with systemic lupus erythematosus.

A parallel testing of antibodies to double-stranded ribonucleic acid in 80 sera from patients with systemic lupus erythematosus by the membrane binding method using natural 3H-dsRNA preparation and synthetic 125I-poly I. poly C preparation revealed a good correlation (r = +0.81). In a selected set of patients with SLE we observed a slight tendency to preferential binding of the natural preparation and a lower frequency of anti-dsRNA antibodies when compared to that reported by other investigators. The presence of anti-dsRNA did not correlate with the presence of anti-dsDNA.

[Study of nucleic acid structure by immunochemical methods. I. Antibodies specific for 6-sulfo-5,6-dihydro-4-methoxyaminopyrimidinone-2]. / Izuchenie struktury nukleinovykh kislot immunokhimicheskimi metodami. I. Antitela, spetsifichnye k 6-sul'fo-5,6-digidro-4-metoksiaminopirimidinonu-2.

Immunization of animals with DNA modified by a mixture of bisulphite and O-methylhydroxylamine and methylated bovine serum albumin results in production of antibodies mainly reacting with modified DNA. Antibodies that react with denatured DNA were produced in minute quantity. It was shown that elicited antibodies possess a high specificity and have the ability to recognize only nucleotides with a double modification. The immune sera were fractionated by Sephadex G-200 column chromatography and the antibody activity was demonstrable in the 19S and 7S fractions. The attempts to induce synthesis of antibodies by injection of DNA modified by O-methylhydroxylamine failed.

[Anion exchange chromatography of proteins leads to induction of DNA-binding activity in them]. / Anionobmennaia khromatografiia belkov privodit k induktsii v nikh DNK-sviazyvaiushchei aktivnosti.

The present work is devoted to investigation of the acquired DNA-binding activity of several proteins, such as bovine serum albumin, myoglobin, and collagens in different conformational states (native, denatured, and gelatin) following their anion exchange chromatography. Induction of DNA-binding activity, as shown, is natural phenomenon common in protein treatment on positively charged matrix. Obviously, anion exchange chromatography of proteins results in their structural conformational changes. It was suggested that the DNA-binding activity of activated proteins is based on the ability of the protein molecule to accept additional protons upon contact with the anion exchanger. This hypothesis is confirmed by the inhibition of the interaction of activated protein with DNA in the presence of Na-EDTA.

[A study of the reaction between antibodies and DNA by the method of immune complex formation on the membranes of nitrocellulose filters]. / Izuchenie reaktsii mezhdu antitelami i DNK metodom fiksatsii immunykh kompleksov na membrannykh nitrotselliuloznykh fil'trakh

The conditions were developed to measure antibodies to denatured and native DNA. It was shown that membrane filters treated with alkali adsorbed denatured DNA complexed with antibodies. Utilizing this method it was shown that antibodies to denatured DNA react with pyrimidines (probably thymine). This assay provides proof of the wide existence antibodies to native DNA in the sera of patients with systemic lupus erythematosus. DNA-binding of sera was significantly depended on reaction condition.

[Antibodies to the thyroid microsomal antigen in children and adolescents subjected to radiation exposure as a result of the accident at the Chernobyl Atomic Electric Power Station]. / Antitela k mikrosomal'nomu antigenu shchitovidnoi zhelezy u detei i podrostkov, podvergshikhsia radiatsionnomu vozdeistviiu v rezul'tate avarii na Chernobyl'skoi AES.

For evaluation of the possibility of the appearance of autoimmune thyroiditis in children and juveniles lived in the areas of Kaluga Province [correction of region] suffered from the Chernobyl accident the content of antibodies to human thyroid microsomal antigen was investigated. Percentage of positive sera varied from 4.8% to 1.2% during 6 years. There is significant difference in the frequency of the antibody appearance between persons suffered from radioactive iodine and unsuffered ones. Correlation between levels of antimicrosomal antibodies and radiation dose obtained from incorporated radioactive iodine was not estimated.

[Immunologic methods in epidemiologic monitoring of population, affected by radioactive iodine as a result of the accident at the Chernobyl nuclear plant]. / Immunologicheskie metody v épidemiologicheskom monitoringe naseleniia podvergshegosiavozdeistviiu radioaktivnogo ioda v rezultate avarii na ChAES.

Ultrasound investigations of the thyroid gland and determinations of microsomal antibodies have been performed in persons who lived in the town of Korosten (Zhitomir Region) during the Chernobyl accident. A high correlation has been found between ultrasound and immunological results. The immunological screening of the population suffered from the Chernobyl disaster might be successfully used for the autoimmune thyroiditis detection. These data complete those obtained by the ultrasound tests.

[The probable sequelae of thyroid damage from radioactive iodine during the Chernobyl accident]. / Veroiatnye posledstviia povrezhdeniia radioaktivnym iodom shchitovidnoi zhelezy v period Chernobyl'skoi avarii.

An important peculiarity of the Chernobyl catastrophe is the discharge into the atmosphere of tremendous amount of radioactive iodine and, as a result, selective damage of the thyroid in children from the affected areas. The most dangerous consequence is the thyroid cancer. The analysis of the situation when children's thyroids were subjected to irradiation shows that tumors can most frequently develop as late as 20-30 years after irradiation. There are reasons to believe that tumors are induced by low dose of irradiation. The most important factor in development of pathologies is for sure the age of the children of the moment of irradiation. A well-known consequence of the impact of radiation on the thyroid is the lymphocyte thyroiditis. The interest to this pathology is determined by the fact that it substantially increases the probability of development of various haematologic diseases (lympho- and myeloproliferative neoplasms).

[Study of the possibility of hyperparathyroidism development in rats exposed to internal irradiation by iodine radioisotope]. / Issledovanie vozmozhnosti razvitiia giperparatireoza u krys, perenesshikh vnutrennee obluchenie ot radioizotopa ioda.

The possibility of hyperparathyroidism development secondary to earlier internal irradiation with radioactive iodine was studied experimentally in Wistar rats. This report describes the parathyroid morphology and biochemical findings for animals irradiated with 131I at the doses of 4.5, 40, or 80 Gy. The interval between the radiation exposure of two-month-old rats and their examination for thyroid and parathyroid pathology was 14 months. Neither hypercalcemia nor hypophosphatemia was found. Moreover, the level of calcium in serum slightly decreased following 40 and 80 Gy irradiation. The increased incidence of parathyroid fibrosis and hypofunctional structure transformation were revealed.

[Determination of the DNA-synthesizing cell count by the immunofluorescence method]. / Opredelenie kolichestva kletok, sinteziruiushchikh DNK, s pomoshch'iu immmunofluorestsentnogo metoda.

In DNA-synthesizing cells DNA is partially single-stranded. Anti-thymidine antibodies, while specifically reacting with this DNA, form a complex which may be revealed using indirect immunofluorescent technique. A comparative determination of DNA-synthesizing cell number in tumor tissue (larynx squamous cell carcinoma) was performed using immunofluorescent technique and radioautography. The former method showed the labeling index (LI) to vary from 1.2 to 9.9%, while the latter showed it to vary from 1.0 to 8.2%. The correlation ratio between the LI values obtained by the two techniques was 0.79. To eliminate a possible reaction of anti-thymidine antibodies with cellular RNA, specimens were preincubated in solutions with RNAase. No more than 6 hours were required to stain specimens using this LI estimation technique. This investigation allows to reveal DNA synthesizing cells not only in the periphery of a histological section, as does routinely radioautography, but also in its centre.

[Transport of 125I-poly I : poly C incorporated in liposomes from the enteric cavity to the small intestine mucosa. An electron microscopic autoradiographic study]. / Transport 125I-poliI : poliTs, zakliuchennogo v liposomy, iz énteral'noi sredy v slizistuiu tonkoi kishki. Issledovanie metodom élektronno-mikroskopicheskoi avtoradiografii.

Transport of 125I-poly(I) : poly(C) incorporated into liposomes trough the small intestine mucose was investigated by electron microscopic autoradiography. With the migration of liposomes into the mucous layer on the luminal surface of the intestine up to the glycocalix level of microvilli these undergo degradation with the formation of monolayer liposomes from which polynucleotide is released. Later on the poly(I) : poly(C) or its fragments transported through the enterocytes to be accumulated in cells of the connective tissue stroma of the small intestine mucose. Part of polynucleotide was incorporated up to the arterial and lymphoid capillary level. Apparently, on the way of its transport the polynucleotide is affected by pancreatic and tissue nucleases. The accumulation of polynucleotide in macrophages, fibroblasts, lymphocytes, plasma cells and smooth muscle cells was traced. It is supposed that the polynucleotide accumulated in stroma of the small intestine mucose may preserve its interferon inducing activity.

[Functional suitability of iodinated casein in the prevention of iodine deficiency]. / Funktsional'naia prigodnost' iodkazeina dlia profilaktiki iodnoi nedostatotochnosti.

At least 1.5 billion in the world and over 100 million Russians live in iodine-deficiency areas. The sequelae of this condition are well known. The paper discusses the properties of the iodinated lactoprotein caseoiodine designed by the MRRC researchers and its capacities and advantages for using in the prevention of iodine deficiency. The assimilation of iodine from caseoiodine corresponds to the body's status: it significantly accumulates more frequently in the thyroid in dietary iodine deficiency (Group 1) and to a lesser degree in adequate dietary iodine intake (Group 2) and, by hour 72, amounts to 36.3 and 31.4% of the ingested amount, respectively (P < 0.05). The findings are in agreement with the data available in the literature on the involvement of hepatic enzymes in regulating the metabolism of iodine during its intake in the protein (casein)-bound form and in increasing its fecal excretion (up to 40% in Group 2) whereas 95% iodine excrete with urine when inorganic iodine is consumed.

[Antibodies to double-helical RNA and several viruses in medical personnel caring for systemic lupus erythematosus patients]. / Antitela k dvuspiral'noi RNK i nekotorym virusam u meditsinskogo personala, obsluzhivaiushchego bol'nykh sistemnoi krasnoi volchankoi.

An average level of antibody to double-stranded RNA in patients with systemic lupus erythematosus and medical staff taking care of these patients is higher than that in physicians from other institutions and donors. The level of antibodies to antigens of various viruses in lupus patients is elevated in comparison with that in the medical staff. No statistically significant differences in the levels of antibody to viruses in the staff of various institutions were found.

[Interferon induction via peroral administration of high-molecular polynucleotides]. / Induktsiia interferona pri peroral'nym vvedenii vysokomolekuliarnykh polinukleotidov.

High molecular polynucleotides, poly(I).poly(C) and poly(G).poly(C), incorporated into liposomes may be used for serum interferon induction when administered orally. Titres of interferon induced by this method are sufficiently high (320-640 units/ml) and close to those induced by the same preparations administered parenterally (640-1280 units/ml). The use of liposomal polynucleotides results in prolonged (up to 24 hours) circulation of interferon in the blood of mice at a sufficiently high level. Liposomal polynucleotides are low-toxic upon oral intake. The liposome-polynucleotide complex is sufficiently stable on storage, and retains the interferon-inducing activity, if given orally, after 6 months of storage at 4 degrees C in the liquid form.

[Interferon-inducing activity of liposome-incorporated double-stranded polynucleotides and the means for its enhancement]. / Interferonindutsiruiushchaia aktivnost' zakliuchennykh v liposomy dvuspiral'nykh polinukleotidov i puti ee povysheniia.

Natural double-stranded RNA (dsRNA) incorporated into liposomes upon parenteral inoculation induces 4 times as much amounts of interferon as inoculation of the equal amount of dsRNA without liposomes. Oral administration of liposome-incorporated dsRNA induces in animals serum interferon in amounts similar to those induced by parenteral inoculation of dsRNA without liposomes (320-640 units/ml). When liposome-incorporated dsRNA is used, interferon induction is prolonged to 24 hours. The prolongation period increases to 5 days after preliminary treatment of animals with "empty" liposomes. In M-19 cell culture, 2-hour treatment with liposome-incorporated dsRNA in a dose of 5-10 micrograms/ml induces a yield of 640-1280 units/ml interferon and 100% antiviral effect. The L-929 culture is more sensitive to dsRNA in liposomes. Even its minimal amounts (0.1-1 microgram/ml) after 2-3-hour contact produce a 100% antiviral effect in the presence of low amounts of interferon in the culture fluid (20-40 units/ml) or in its complete absence.

[Characteristics of interferon production from the enteral administration of inducers]. / Osobennosti produktsii interferona pri énteral'nom vvedenii induktorov.

When interferon inducers are administered to animals, autonomous local production of interferon by organs (intestine, liver) occurs alongside with the production of serum interferon. After oral administration of a low molecular interferon inducer, tilorone, the intestine is the main site of interferon synthesis. Here the peak of interferon production precedes that in the serum by 20 hours and exceeds it 16-fold. The sequence of events follows the scheme: intestine--liver--serum. Upon oral administration of high molecular inducers incorporated into liposomes (polyguacyl, lafarin) the predominant site of interferon production is the liver where over 80% of the administered inducer is localized and interferon production is 2-4-fold higher than in the intestine. The sequence of events follows the scheme: liver--intestine--serum.

[DNA-binding proteins in human serum]. / DNK-sviazyvaiushchie belki syvorotki krovi cheloveka.

Fractions of acid and base blood serum proteins, separated by ion exchange chromatography on QAE-Sephadex (but not the proteins of the whole blood serum, which had the same charge), reacted with DNA preparations. Separate fractions of blood serum proteins were able to react with DNA after electrophoretic separation. Binding of blood serum proteins with DNA did not depend on ion strength within the range of NaCl concentration from 0.1 M to 0.5 M; it was also stable at pH 5.5 = 8.0. Interaction between acid DNA-binding proteins and native DNA was inhibited by denatured DNA, polyguanilic and polyinosinic acids and by other polyanions: dextran sulfate, polyvinyl sulfate, polyanetol sulphonate, polystyrol sulphonate and heparin. Acid DNA-binding proteins showed only a slight affinity to polyadenilic, polyuridilic and polycytidilic acids. The acid and base DNA-binding proteins appear to be contained in blood serum in the form of a loosely bound complex.

[Spontaneous secretion of antinuclear factors in systemic lupus erythematosus, rheumatoid arthritis and rheumatism]. / Spontannaia sekretsiia antinuklearnykh faktorov pri sistemnoi krasnoi volchanke, revmatoidnom artrite i revmatizme.

The authors studied the ability of non-stimulated short-term cultures of peripheral lymphocytes of SLE, RA and rheumatic fever patients to spontaneously produce antibodies to DNA and other antinuclear factors. To demonstrate antibodies to DNA, use was made of the radioimmunoassay and the passive hemagglutination test. ANF-test of indirect immunofluorescence. The nosological and clinical features of secretory ANF were explored in the diseases in question. It was established that patients with the main rheumatic diseases are different as regards the rate of demonstration, level and spectrum of secretory ANF. They also differ from the group of donors in terms of the same characteristics. The culture of peripheral lymphocytes of SLE patients has the greatest ability to secrete antibodies to DNA and ANF of the primarily marginal and homogenous types of fluorescence. This characteristic correlates with SLE activity before the disease onset and over time as well as with lupus nephritis. In RA and rheumatic fever, this characteristics correlates with the disease activity, the presence of visceritis in RA. The clinicopathogenetic importance of secretory ANF in rheumatic diseases is discussed.

[A simple radioimmunological method of determining thymidine in biological fluids]. / Prostoi radioimmunologicheskii metod opredeleniia timidina v biologicheskikh zhidkostiakh.

A simple, sensitive and reproducible radioimmunoassay for determination of thymidine in biological fluids was developed. The technique is based on competition between 3H-thymidine and unlabelled thymidine for antithymidine antibodies. The cross-reactivity of structurally related compounds with the antibody was mostly negligible, except for deoxyuridine. The sensitivity of the assay was 10(-7) M. This radioimmunoassay may be used for measuring content of thymidine in blood serum and urine. A method for direct determination of thymidine in urine was not described before.