Genetics and evolution of the Lpm-system in American mink. VI. Identification, genetic control and phylogenesis of the new allotype Lpm13

Data on immuno- and biochemical identification, genetic control and phylogenesis of new allotype Lpm13 of the Lpm system in domestic mink are presented. This allotype is encountered in mink populations with the frequency 0.9 and higher. The availability of Lpm13 genetic marker permitted another haplotype to be revealed, in addition to the eight known Lpm haplotypes by means of genetic analysis. It was established that, alongside with the earlier described haplotype Lpm3,4,6,8,9,10,11 (abbreviation H3), there exists a similar haplotype, Lpm3,4,6,8,9,10,11,13 (abbreviation H3.13), containing the Lpm13 gene. Of the rest seven haplotypes, five have the Lpm13 gene and two do not. Taking into account this gene and corresponding antigenic marker, the differentiation of 28, instead of 25, phenotypes and 45, instead of 36, genotypes for the Lpm system became possible. Lpm13 antigenic specificity was found with no exception in all individual serum samples taken from ten species and interspecific hybrids of Mustelidae which are closely related to domestic mink. The data obtained give grounds to refer the newly identified Lpm13 gene to the first evolutionary conservative category of genes of the multigenic Lpm system which is also represented by the Lpm6, Lpm9, Lpm10 and Lpm11 genes. The hypotheses of instantaneous formation of polymorphism of the Lpm system in domestic mink are briefly regarded.     

Localization of the α2-macroglobulin gene and Lpm gene family on mink chromosome 9

Summary Using cloned cDNA for human ₂-macroglobulin (A2M) as a probe, mink-Chinese hamster hybrid cells were analysed. The results allowed us to assign a gene for A2M to mink chromosome 9. Breeding tests demonstrated that the Lpm-locus coding for other related -macroglobulin protein and the gene for peptidase B (PEPB) are linked 11±3 cm apart. The PEPB gene is located on mink chromosome 9, and hence, the Lpw-locus is on the same mink chromosome. The relationship of the genetic systems controlling the isotypically different -macroglobulins in mink serum are discussed.     

Common antigens of mouse oval and biliary epithelial cells. Expression on newly formed hepatocytes

Two antigens - A6 and G7 - shared by mouse biliary epithelial and oval cells were revealed by monoclonal antibodies raised in rat immunized with oval-cell-enriched liver fraction. Oval cells were induced in CBA or F1 (CBA x C57BL6) mice by a combination of a single injection of the alkylating drug Dipin with partial hepatectomy. In normal liver A6 antigen was localized, using light and electron microscopy, in biliary epithelial cells of all ducts including Hering canals. Some bile ductal and Hering cells were A6-negative. Occasionally, A6 antigen was present in single hepatocytes forming the periportal ends of hepatic cords. In preneoplastic and tumorous liver A6 antigen was present in bile ductal and oval cells and in a fraction of newly formed hepatocytes and tumor cells. G7 antigen was revealed in normal, precancerous and tumorous liver in biliary epithelial and oval cells but not in hepatocytes. A6 and G7 antigens were not liver-specific: they were expressed in various normal organs and tissues, especially in epithelia. In studies of mouse liver lineages A6 antigen can be used as a common marker of biliary epithelial and oval cells and hepatocytes at certain stages of differentiation. G7 antigen is a marker of oval and biliary epithelial cells. There was a striking similarity in A6 antigen localization to that of human blood group antigens in normal liver and liver tumors. A6 antigen may thus provide a useful tool for the study of neoexpression of human blood group antigens in liver tumors.     

Structural analysis of translating ribosomes. Proton magnetic resonance method

Comparison has been made of the proton magnetic resonance (PMR) spectra of translating ribosomes in the pre-translocation and post-translocation states as well as of the complexes of translating ribosomes with elongation factors Tu (EF-Tu) or G (EF-G) in the presence of the uncleavable analogue of GTP--guanylyl-imidodiphosphate (GMP-PNP). It is shown that proteins L7/L12 within the translating ribosomes possess a high intramolecular mobility both in the pre-translocation and in the post-translocation states. The interaction of EF-G with translating ribosomes results in a decrease of the mobility of the L7/L12 proteins. The interaction of EF-Tu with translating ribosomes leads to slight changes in the PMR spectra different from the changes caused by EF-G.     

Effect of ascorbic acid prophylaxis on the frequency of chromosome aberrations, urine mutagenicity and nucleolus test in workers occupationally exposed to cytostatic drugs

The genetic risk of workers occupationally exposed to a series of newly developed cytostatic drugs and the presumed antimutagenic potential of ascorbic acid (AA) were studied in a group of 38 chemical laboratory personnel examined for chromosome aberrations in lymphocytes, urine mutagenicity and nucleolar RNA activity before and after a 6-month prophylactic administration of AA at daily doses of 1 g for 5 days a week. Chromosome aberration tests revealed elevated aberrant cell (AB.C) rates both prior to and after AA supplementation (3.9% and 3.65% of AB.C., respectively). These values were significantly higher than those found in 18 non-exposed matching controls (1.05% of AB.C.). Tests for mutagenic activity in the urine of drug-exposed workers revealed 64% positive urine samples prior to vitaminization and 60% positive urine specimens after it; positive urine samples in the group of controls accounted for 21% of samples. In the nucleolus test, numbers of inactivated micronuclei in the exposed were initially higher than those of controls (33.4% versus 24.3%), but dropped to 20.5% after AA supplementation. These findings show that AA prophylaxis alone cannot substantially reduce the hazards associated with exposure to anti-cancer drugs.     

Staphylococcus aureus at a maternity ward. II. Characterization of isolates by enterotoxigenicity and toxic shock syndrome toxin (TSST-1) production

Tests for enterotoxins A, B, C, D, E and TSST-1 production were carried out on 775 S. aureus strains isolated from various sources (50 mothers and neonates studied periodically, mothers and infants treated for various acute inflammatory conditions, members of hospital staff, environmental swabs) during the period 1981-1983 at a maternity ward chosen for a 3-year systematic study and on additional 97 isolates obtained in 1985 from another maternity ward. This had contributed to a better classification of strains within certain phage type groups. It was found that the distribution of S. aureus types in the particular sub-sets varied, depending on the source of isolates. At the maternity ward followed for 3 years there was a clear-cut trend towards the spread of phage-untypable isolates producing enterotoxin C whereas at ward examined for comparative purposes B enterotoxin producers of phage type 95 were predominant. The tests for enterotoxigenicity has also proved to be useful as the epidemiological marker characterizing the predominantly circulating S. aureus strain. It has been confirmed that the majority role in the spread of maternity-ward-staphylococci is played by the neonates and the factors of hospital environment.     

Prevention of infectious complications in patients with acute bone marrow depression]

Clinical efficacy of intravenous gentamicin in combination with oral use of gentamicin, ristomycin and nystatin was studied in 1977. In 1980-1984 two antiinfectious regimens were tested: intravenous administration of gentamicin in combination with total decontamination (oral use of gentamicin and nystatin) and selective decontamination (biseptol with nystatin of amphoglucamine). It was shown that the incidence of severe infections in the patients under the observation dropped against the controls. The incidence and spectrum of infectious complications in cases with acute myelodepressions were proved to depend on the intestinal autoflora inhibition.     

Cloning of the DNA fragment of a transgenic mouse containing an integrated recombinant plasmid

The "plasmid rescue" method has been used to isolate the recombinant plasmid pMA3 fragment and flanking sequences from the transgenic mouse genome containing the fragment in integrated form. The "rescued" plasmid, pMAR1, lacks all virus sequences and retains only those regions of pBR322 that are responsible for the plasmid replication and Escherichia coli ampicillin resistance. The plasmid pMA3 deletion has occurred at its integration into the mouse genome after microinjection into the zygote. The integrated fragment of the plasmid is adjacent to the genome repeated sequence that is highly conservative in evolution.     

Structural studies of translating ribosomes. II. Comparative sedimentation analysis of pre- and post-translocation states

Using the solid-phase translation system technique where template poly(U) is covalently coupled to Sepharose through cleavable disulfide bridges translating monoribosomes carrying a polypeptide (polyPhe) of 10 to 20 amino acids long have been isolated. Both pre-translocation state and post-translocation state ribosomes have been obtained. It has been shown that the sedimentation coefficient of the pre-translocation state ribosomes exceeds that of the post-translocation state ribosomes by a magnitude of about 1S. This difference is independent on the sedimentation rate (hydrostatic pressure) in the range of 20 000 to 40 000 rev/min and, most likely, is not a direct contribution of the increase of the particle mass at the expense of an additional tRNA in the pre-translocation state ribosomes. Together with other data, this result suggests that translating ribosomes in the pre-translocation state are more compact than post-translocation state ribosomes.     

Characteristics of ceruloplasmin synthesis in mammalian embryogenesis. 2. The yolk sac as the site of the primary expression of the ceruloplasmin gene in rats

It was shown that the omphaloid placenta and, first of all, visceral wall of yolk sac is the site of primary synthesis of ceruloplasmin (CP), whereas the activation of CP synthesis in the liver cells is secondary and is revealed from the 12th day of embryo-genesis. The CP synthesis in the yolk sac cells proved by selective CP localization in the cells of the yolk sac visceral wall and, first of all, in the cells of visceral endoderm on sections stained by the method of indirect immunofluorescence and using the reaction of soluble peroxidase-antiperoxidase complex. A specific CP-mRNA has been revealed in the yolk sac cells which is actively translated in the polyribosomes isolated from the yolk sac and in the cell-free translation system from the rabbit reticulocytes. on the 14th day of embryogenesis CP amounts to ca. 4% of all polypeptides secreted by the yolk sac cells. As the embryogenesis proceeds, the relative rate of CP synthesis progressively decreases in the yolk sac and increases in the liver cells. CP synthesized by the yolk sac cells has a molecular mass of ca. 122 kD. Possible causes of differences between the "embryonic" and "adult" rat CPs are discussed. A suggestion has been put forward that the time of activation of CP synthesis coincides with the yolk sac formation (8-9th days of embryogenesis) and the cells of visceral endoderm are the site of primary expression of the CP gene.