PHYSIOLOGICAL, CYTOLOGICAL AND BIOCHEMICAL STABILITY OF Medicago sativa L. CELL CULTURE AFTER 27 YEARS OF CRYOGENIC STORAGE.

BACKGROUND: The efficiency of long-term cryogenic storage to prevent somaclonal variations in plant cell cultures and retain their major cytogenetic and biochemical traits remains under debate. In particular, it is not clear how stress conditions associated with cryopreservation, such as low temperature, dehydration and toxic action of some cryoprotectants (DMSO in particular), affect post-storage regrowth and genetic integrity of cell samples. OBJECTIVE: We assessed growth, cytogenetic and biochemical characteristics of the peroxidase-producing strain of Medicago sativa L. cell culture recovered after 27 years of cryogenic storage as compared to the same culture before cryopreservation. MATERIALS AND METHODS: In 1984, M. sativa L. cell culture was cryopreserved using programmed freezing and 7% DMSO as a cryoprotectant. In 2011, after rewarming in a water bath at 40 degree C for 90 s, cell culture was recovered and proliferated. Viability, growth profile, mitotic index, ploidy level, peroxidase activity and cell response to hypothermia and osmotic stress were compared between the recovered and the initial cell cultures using the records available from 1984. RESULTS: Viability of alfalfa cell culture after rewarming was below 20% but it increased to 80% by the 27th subculture cycle. Recovered culture showed higher mitotic activity and increased number of haploid and diploid cells compared to the initial cell line. Both peroxidase activity and response to abiotic stress in the recovered cell culture were similar to that of the initial culture. CONCLUSION: Cryopreservation by programmed freezing was effective at retaining the main characteristics of M. sativa undifferentiated cell culture after 27 years of storage. According to available data, this is longest period of successful cryopreservation of plant cell cultures reported so far. After storage, there was no evidence that DMSO had any detrimental effect on cell viability, growth or cytogenetics.

[Adaptogenic effects of furostanol glycosides of Dioscorea deltoidea wall on oxidative processes in tomato plants in biotic stress].

The effect of furostanol glycosides of cell culture of Dioscorea deltoidea Wall on oxidative processes in tomato plants subjected to invasion with the gall nematode Meloidogyne incognita Kofoid et White was studied. We showed that furostanol glycosides induce a nonspecific defensive response in plants. Exposure of cell membranes to furostanol glycosides cause rearrangements in fatty acids resulting in the formation of conjugated dienes, which makes molecules thermodynamically more stable under stress conditions. The study of changes in the activity of peroxidases of intact plants and plants affected with the nematode, which were treated with furostanol glycosides, showed that the protective effect of the guaiacol-dependent peroxidase is more long-term than the effect of the benzidine-dependent peroxidase.

[Impact of the impaired intestinal microflora on the course of acne vulgaris]. / Vliiane disbakterioza kishechnika na techenie vul'garnykh ugrei.

The paper deals with studies of the intestinal microflora in 114 patients with acne vulgaris (94 and 20 with its papulopustular and nodulocystic forms). Sixty-one (54%) patients have either the first (21%) or second (78.7%) impaired bacterial microflora. At the same time, there are no great differences in the content of the intestinal microflora in different forms of acne. It is noted that adding intestinal microflora-correcting agents to combined therapy in patients with papulopustular acne vulgaris and verified dysbacteriosis reduces the duration of treatment by over twice and makes its duration the same as that in patients without dysbacteriosis.

Study of the structure of streptokinase conjugates with a hydrophilic vinylpyrrolidone copolymer.

The modification of streptokinase by a synthetic N-vinylpyrrolidone copolymer leads to formation of conjugates varying in structures according to the proportions of the components in the reaction medium. Based on data obtained from spectrophotometry, as well as sedimentation and diffusion analyses, it is shown that in the presence of excess protein in the reaction medium, formation of the main chain takes place via the copolymer associated with several protein globules. Under conditions of excess modifier copolymer, either single-site and/or multiple-site bonding is possible for the protein backbone, depending on the molecular weight of the copolymer. One of the models for the conjugates obtained in this manner has been corroborated by small-angle X-ray scattering data. CD spectral analyses has been performed in order to demonstrate that covalent modification does not alter the secondary structure of streptokinase in the conjugate whereas the tertiary structure undergoes local changes in conformation.

[Use of non-traditional methods of examination in the differential diagnosis of cancer and tuberculosis of the lungs]. / Ispo'lzovanie netraditsionnykh metodov issledovaniia v diffe- rentsia'lnoi diagnostike raka i tuberkuleza legkikh.

The immunoradiometric assay of the blood serum was performed in 61 patient with cancer and in 57 patients with tuberculosis of the lungs to determine the content of adrenocorticotropic hormone (ACTH), hydrocortisone, triiodothyronine++ (T3), thyroxin (T4), ferritin and carcinoembryonic antigen (CEA). In 200 patients (104 suffered from pulmonary tuberculosis, 9--pulmonary cancer), entropy of the protein fractions of the blood serum was determined. It was established that in differential diagnosis of pulmonary cancer and tuberculosis, the determination of CEA, hydrocortisone and entropy of the protein fractions of the blood serum in the complex with the other methods were of significant importance, the determination of ACTH and ferritin content were less important.

[A study of native streptokinase and its polymeric derivative by means of small angle x-ray scattering]. / Issledovanie nativnoi streptokinazy i ee polimernogo proizvodnogo metodom rentgenovskogo rasseianiia pod malymi uglami.

The gyration radius (R0) of native streptokinase (SK) was found to be R0 = (40 +/- ) A by small-angle X-ray scattering. Experimental hydrodynamic characteristics of SK were S0(20),W = (2.8 +/- 0.1)S; D0(20),W = (6.0 +/- 0.5) x 10(-7) cm2/s; [n] = 0.12 dl/g. The molecular weight of the enzyme was found to be 44,000. The values of the form factor R0/Rsphere = 2.1 and the frictional ratio f/f0 = 1.5 indicate considerable anisometry of the SK molecule. Basing on the curves of small-angle X-ray scattering of SK modified with a synthetic linear copolymer of N-vinylpyrrolidone (P) at a molar ratio SK less than P, a structural model of the conjugate was proposed. The modified form consisted of a dense nucleus covered with a diffuse polymeric membrane. In accordance with the model, R0 of modified SK and of the whole conjugate were found to be R0nucleus = (34 +/- 2) A and R0conjugate = (114 +/- 5)A.

[Excretory function of the liver with an allyl alcohol lesion and the antioxidant correction of the disorders]. / Ekskretornaia funktsiia pecheni pri porazhenii allilovym spirtom i korrektsiia narushenii antioksidantami.

In experiments on 33 albino male rats it was established that during allyl alcohol-induced damage of the liver its excretory function is impaired that was judged by the biliary excretion of intravenously injected Bengal pink-131I. Antioxidants (tocopherol acetate, sodium selenite, unithiol) reduced the toxic effect of allyl alcohol on the liver.

[Prevention of post-abortion complications]. / Profilaktika posleabortnykh oslozhnenii.

PIP: A study was conducted on the effectiveness of vibratory massage of the uterus in preventing post-abortion complications. Observations were made on 303 pregnant women, ages 20-40, admitted to the gynecology clinic for induced abortions. The base group consisted of 168 women who received vibratory massage of the uterus after the abortion; the control group consisted of 135 women not given vibromassage. The base group included 28 (17%) primiparae, 9 (5.3%) parae and 27 (17%) nulliparae; 60 (36%) had each had 3 abortion, and 27 (17%) had had 4 or more; 43 (25.5%) of the women had their fit menstruation by the age of 15 and later; painful menstrual function (algodysmenorrhea) was noted in 44 (26%) of the women; 41 (32%) had sex for the first time at an early age (14-16 yrs). Histories showed chronic inflammation of adnexa for 20 (12.5%) and cervical erosion for 6 (3.5%). Similar data were obtained in the control group. In the base group, 28 of the women were 6-8 weeks pregnant, 92 were 8-10 weeks and 48 were 11-12 weeks, in the control group, the numbers were 44, 56 and 35, respectively. Induced abortions were performed by curettage (126 in the base group, 110 in the control group) and vacuum-aspiration (42 and 25, respectively). Vibromassage of the uterus was performed immediately after the abortions and on the 1st and 2nd days of the post-abortion period. The effect of the vibromassage was evaluated by clinical observation of the women, uterine probing, amount of blood loss, examination of vaginal discharge for microflora, clinical blood analysis, and immediate and late post-operative complications. In the group with abortions preformed by vacuum-aspiration with vibromassage, no heavy blood loss was detected, and the number of women with little or no discharge increased significantly compared to the control group. In the group receiving vibromassage, hematometra was detected in 2.4% of the women, endometritis on 6%, exacerbated chronic inflammation of adnexa in 4%. In the control group, the figures were 6, 13.3 and 17.6%, respectively. In observations for 12-24 months after the abortions, among 100 women who received vibromassage, disturbance of menstrual function was noted in 19.3% and sterility in 8.2%, while in the control group 14.1 and 16.2%, respectively. In the first 6 months after abortion, women not using contraception became pregnant in 7% of the cases, and in 9.1% of the cases among those who received vibromassage. Thus, studies showed that vibratory massage of the uterus, employed immediately after the induced abortion and the next two days of the post-abortion period, has a stimulating effect on the contracting activities of the uterus and contributes to a decrease in immediate and late complications.^ieng

[Seasonal features of the excretory function of the liver in the presence of tetracycline damage and correction of the disorders with antioxidants]. / Sezonnye osobennosti ékskretornoi funktsii pechni pri tetratsiklinovykh porazheniiakh i korrektsiia narushenii antioksidantami.

It was shown on pubertal albino rats that the intensity of excretion with the bile of radioactive Bengal rose was different at different seasons: the maximum and minimum levels were observed in winter and summer, respectively. When the liver was affected with tetracycline, this process was suppressed especially in summer. The use of antioxidants, such as tocopherol acetate in combination with sodium selenite promoted the recovery of liver excretion function in winter, spring and autumn. In summer, the recovery was only partial.