Characteristics of continuous cell line 293 used for preparation of antineoplastic agent Cancerolysin]

Certification of continuous cell 293 culture used for cultivation of antineoplastic preparation Cancerolysin was carried out. The seeding and working banks of cells 293 were established and deposited for storage at the Vector Centre. The cells were certified in accordance with the WHO requirements. The cell 293 culture was shown to have high proliferative activity; morphology typical of the line; its karyotype and enzymogram are typical of human cells; the culture is not contaminated with bacteria, fungi, mycoplasms and viruses including oncogenic ones; it has high virus-producing activity; it preserves stability of all the biological properties in long-term cultivation. The seeding and working cell banks were recommended for the use in production of drugs for the treatment of oncologic patients.     

Real-time ultrasound imaging for predicting ovine fetal age

Fetal head width and thoracic depth were observed via real-time ultrasound imaging in 12 single-, 12 twin- and 4 triplet-bearing BooroolaxSouth Australian Merino ewes at weekly intervals during mid pregnancy. Fetal age varied by a maximum of 24 h. Relationships between fetal age and the linear measures (head width, thoracic depth) were determined within litter size categories, using both linear and quadratic terms. All relationships tested for linearity were statistically significant (P<0.05). A curvilinear relationship (P<0.05) was observed for fetal age and thoracic depth of twin fetuses. Significant (P<0.05) curvilinear relationships were also observed for fetal age and head width, and fetal age and thoracic depth when litter size data were pooled. We conclude that head width and thoracic depth are acceptable linear measurements for estimating fetal age in the ovine species. Implications of these results for research and for the commercial field are discussed.     

Synthesis and high stability of complementary complexes of N-(2-hydroxyethyl)phenazinium derivatives of oligonucleotides.

Two simple methods for the synthesis of oligonucleotides bearing a N-(2-hydroxyethyl)phenazinium (Phn) residue at the 5'- and/or 3'-terminal phosphate groups are proposed. By forming complexes between a dodecanucleotide d(pApApCpCpTpGpTpTpTpGpGpC), a heptanucleotide d(pCpCpApApApCpA), and Phn derivatives of the latter, it is shown that the introduction of a dye at the end of an oligonucleotide chain strongly stabilizes its complementary complexes. The Tmax and the thermodynamic parameters (delta H, delta S, delta G) of complex formation were determined. According to these data, coupling of a dye with the 5'-terminal phosphate group is the most advantageous: delta G(37 degrees C) is increased by 3.59 +/- 0.04 kcal/mol compared to 2.06 +/- 0.04 kcal/mol for 3'-Phn derivatives. The elongation of the linker, which connects the dye to the oligonucleotide, from a dimethylene up to a heptamethylene usually leads to destabilization of the oligonucleotide complex. The complementary complex formed by the 3',5'-di-Phn derivative of the heptanucleotide was found to be the most stable among all duplexes investigated. Relative to the unmodified complex the increase in free energy was 4.96 +/- 0.04 kcal/mol. The association constant of this modified complex at 37 degrees C is 9.5 x 10(6) M-1, whereas the analogous value for the unmodified complex is only 3 x 10(3) M-1.     

The characteristics of the skeletal muscle fibers of the m. vastus lateralis in highly qualified skaters (a morphometric analysis)

Morphometrical characteristics of the skeletal muscle fibers of the m. vastus lateralis has been presented in the skaters of a high qualification. The muscle biopsies have been obtained by means of the needle biopsy method from 5 sprinters and 10 classical all-round competitors. Longitudinal sections of the peripheral and central parts of the muscle fiber have been investigated, subdividing the fibers in accordance with certain ultrastructural criteria into two types: slow and fast. The area of mitochondria, myofibrils, lipid inclusions has been determined by means of the system of the image analysis ACM-68K (Leitz, BRD). To the defining factors in the skater muscle ultrastructure, influencing the level of sporting achievements, a high parameter of the volumetric density of the whole mitochondrial population (7.6% in the sprinters and 9.2% in the all-around competitors) and an exceptionally high parameter of the sarcoplasmic density (28.5% in the sprinters and 36% in the all-around competitors) should be ascribed. The morphometrical data of the skaters muscle fiber are compared with the muscle parameters (according to the literature data) of the m. vastus lateralis in high qualification sportsmen of other specialization and in nontrained persons.     

Efficacy of therapeutic and prophylactic actions of ultralow doses of antibodies to gamma-interferon in experimental murine model of herpes virus]

The process of the disease due to herpes simplex virus types 1 and 2 (HSV-1 and 2) was studied on white uninbred mice weighing 10 to 12 g. The animals were infected intracerebrally or intraperitoneally. Intraperitoneal contamination of the animals with MS strain of HSV-2 was used for the experimental model of the herpes simplex infection. The prophylactic antiherpes action of ultralow doses of the human gamma-interferon antibodies (ULD of anti-IFN-gamma) at a course of its intragastral administration was evaluated. The preparation was shown to have a significant (p < 0.05) protective effect in a dose of 10 LD50, evident from a 10-fold decrease of the HSV-2 accumulation in the brain, a lower percentage of the animal deaths and an increase of the average lifespan of the animals by 3.3 days. The study of the therapeutic action of ULD of anti-IFN-gamma at a course of its intragastral administration showed that the preparation had no significant positive effect on the disease process in the animals infected with HSV-2 in a dose of 10 LD50. However, a positive effect associated with delayed virus replication in the brain was observed in the study on the therapeutic effect of ULD of anti-IFN-gamma after its intragastral administration to the mice infected with a sublethal dose of the virus.     

Cerebral perfusion changes in patients with internal carotid artery stenoses after surgical revascularization

Changes of cerebral perfusion and the condition of collateral blood supply in patients with internal carotid artery stenoses may have a prognostic value for effective blood flow restoration after revascularization of the internal carotid arteries (ICAs). To determine the patterns of cerebral perfusion changes in patients with ICA stenoses before and after surgical treatment, a clinical CT perfusion study of 41 patients with moderate to severe ICA stenoses was performed. Perfusion CT (PCT) had been conducted in 17 patients with moderate ICA stenoses (50–69%) and in 24 patients with severe ICA stenoses (70–99%) 3 times: before intervention (balloon angioplasty with stenting or carotid endarterectomy), on the 3rd to 7th day, and within 1 to 3 months after surgery. Scanning was performed at the level of the basal ganglia and semioval centers. In patients without ICA stenosis (control group of 39 individuals), PCT was conducted once. We found that surgical recanalization of the ICA leads to normalization of the cerebral blood flow parameters in the perfusion area of the middle cranial artery, as evidenced by a decrease of MTT and CBV and an increase of CBF to values comparable to those in the control group. However, blood flow restoration in the anterior and posterior watershed areas, which are known to be mostly affected under chronic hypoperfusion conditions, was observed only in patients with a complete circle of Willis (CW) and moderate ICA stenosis. Therefore, severe stenosis (>70%) and the incomplete CW are the prognostic factors for inadequate blood flow restoration after revascularization in patients with ICA stenoses.     

Daphniopsis pusilla Serventy (Cladocera: Daphniidae), an important element in the fauna of Australian salt lakes

Daphniopsis pusilla Serventy, an endemic species of cladoceran found in Australian salt lakes, is redescribed. It occurs in both the south-west and south-east of Australia (including Tasmania), and at salinities between about 2 and 71‰     

Structural and kinetic aspects of chemical reactions in DNA duplexes. Information on DNA local structure obtained from chemical ligation data.

Chemical ligation of oligonucleotides in double-stranded helices has been considered in its structural-kinetic aspect. A study was made of (i) two series of DNA duplexes with various arrangements of reacting groups in the ligation junction induced by mispairing or by alteration of furanose structure (the replacement of dT unit with rU, aU, IU, xU, dxT ones) and of (ii) eight synthetic water-soluble carbodiimides with different substituents at N1 and N3 atoms. We assumed that some information on the local structure of modified sites in the duplex can be obtained from kinetic parameters of oligonucleotide coupling reaction. The ratio of kinetic constants k3/(k2 + k3) for productive and nonproductive decomposition of the activated phosphomonoester derivative apparently reflects the reaction site structure: for a given duplex this parameter is virtually independent of the condensing agent composition. Based on the analysis of the chemical ligation kinetics a suggestion has been made about the conformation of some modified units in the double helix.     

The affected sib-set method: revision based on the mixed model using a conditional probability approach]

One of the implicit assumptions of the single locus model, having been used so far in the analysis of linkage between the genetic marker locus and the disease predisposition locus, is the requirement of independent--from the rest of genotype--action of genotypes of the disease predisposition locus considered. In this communication, it is emphasized that the lack of this requirement makes problematical the theoretical substantiation of the affected sib-pair method in the linkage analysis. To remove this obstacle, explicit pointing out of independence of the action of the single locus genotypes on the rest of the genotype is necessary in formulating of the single locus model which, with due regard for this assumption, represents a special, perhaps, unique case of the gene action characterized by incomplete differential penetrances of the genotypes under conditions, when the genes of the rest of genotype involved to the disease, are fixed. In this connection, the mixed model of inheritance with the "major gene", proposed by Morton and MacLean (1974), is considered, on the basis of which the theoretical expectations of the proportions of the affected sib pairs, sharing the x = 2, 1, 0 haplotypes, identical by descent (IBD) in phenotypic matings with the h = 2, 1, 0 affected parents are derived. Based on the combinatorial analysis of IBD relationships in sib pairs and of the distribution of sibships of any size s greater than or equal to 2 by the numbers L = 2, 3, 4 haplotypes, inherited by s siblings, the empirical assessment of data on sibships of any size with r greater than or equal to 2 affected siblings is considered, which makes it possible to reduce the data observed on distribution of the numbers L in sibships, to that of the IBD relationships in the affected sib pairs. It is also pointed out that conditional probability approach, proposed by the author earlier, allows at the same time to obtain the empirical estimates of the recurrence risks, conditional both on phenotypes of siblings (r affected; s-r normal siblings), and on the number of L haplotypes inherited by sibships.