[Early activation after orthotopic heart transplantation].

The purpose of the study was to analyze the authors' experience with early activation made in the operating-room in recipients after orthotopic heart transplantation (OHT). The study enrolled 12 patients (10 males and 2 females) aged 18 to 60 (41 +/- 5) years. OHT emergency corresponded to UNOS status 2 (n=9) and IB (n=3). The donors' age was 21 to 44 (35 +/- 3) years; the donor/recipient body weight ratio was 0.87 +/- 0.14; the least dopamine-induced cardiotonic support during conditioning was 4.7 +/- 0.4 microg/kg/min. The duration of myocardial ischemia was 106 to 237 (161 +/- 19) min. That of extracorporeal circulation (EC), surgery, and anesthesia was 66 to 129 (99 +/- 8) min, 4.5 +/- 0.3 and 6.2 +/- 0.5 hours, respectively. The induction of anesthesia used propofol 1.6 +/- 0.2 mg/kg, fentanyl 3.8 +/- 0.3 microg/kg, or rocuronium bromide 1.1 +/- 0.2 mg/kg. The maintenance of anesthesia in the preperfusion period was made with propofol (target concentration, 1.5 +/- 0.3 microg/ml), sevoflurane (1.4 +/- 0.3 vol %), and fentanyl. The mean use of propofol per surgery was 2.3 +/- 0.3 mg/kg/h; that of fentanyl and rocuronium bromide was 16.9 +/- 0.6 and 1.7 +/- 0.3 mg/ kg, respectively. In all recipients, the early posttransplantation period was characterized by stable central hemodynamics. At the end of surgery, cardiac index was 3.2 +/- 0.4 l/min/m2 during cardiotonic therapy: epinephrine 42 +/- 8 ng/kg/min (n=10), dopamine 6.4 +/- 0.7 microg/kg/min (n=12), or dobutamine 4.1 +/- 0.4 microg/kg/min (n=3). The interval between the end of surgery and tracheal intubation was 53 +/- 8 min. The duration of postoperative treatment in an intensive care unit was 2 to 4 (2.8 +/- 0.5) days. Thus, early activation can be made on the operating table after heart transplantation if there is no significant cardiac graft pump dysfunction or homeostatic disorders in recipients.

[Biocenosis sparing treatment in the surgery of gastro-intestinal tract].

The authors present results of investigation of the state of microbiocenosis of the intestine and immune status of organism of 139 patients after emergency operations on the gastro-intestinal tract organs. Disbiosis and reduced indices of the immune system in the postoperative period was a cause of using biocenosis sparing treatment resulting in stabilization of microecology of the gastro-intestinal tract, higher colonization resistance and general reactivity of organism.

[Correction of NO-dependent pulmonary vasodilation disorder in cardiosurgical patients with acute respiratory distress syndrome].

Disturbances in endogenous NO synthesis present an important pathogenetic mechanism of NO-dependent vasodilation disorder in patients with acute respiratory distress syndrome. Inhalational NO is an effective means of correcting NO-dependent pulmonary vasodilation and the pulmonary function of oxygenation in 47% of patients with acute respiratory distress syndrome.

[Surgical treatment of bilateral hernias of the inguinofemoral area].

Consecutive surgical treatment of bilateral hernias leads to the appearance of recurrences in 67% of the operated patients. An important etiological moment of the appearance of the repeated, by turns recurrences in the two-steps elimination of a bilateral inguinal hernia is the symptom of tension and pulling the lateral muscles of the abdomen from one side to the other. The proposed method of a one-step surgical treatment, using the suprapubic access by the developed technique, allows reduction of recurrences to 2.2%.

[Milk ceruloplasmin as a physiological source of nutritional copper in early ontogenesis in mammals].

This work elucidates the role of ceruloplasmin (Cp): the main soluble copper containing glycoprotein, in newborn infant copper metabolism. An over 3-fold drop of Cp and copper concentration in samples of skimmed milk from 2 to 5 days of lactation was demonstrated. It has been shown that [125I]Cp of the breast milk is discovered in the blood plasma after its was administered per os to 6-day old rats. In the body, [125I]Cp was specifically bound to cells and lost copper ions. Alternatively, after copper metabolism change to adult type, milk [125I]Cp isn't carried from gastrointestinal tract to the blood. During 8 days, newborn rats were fed with baby formula and we have found that in this case changing of the copper metabolism type takes place earlier in contrast to the litter fed by females. Transcription of the Cp gene activated in liver, Cp and the copper ions concentration in the blood plasma increased proportionally, whereas in liver the copper concentration is decreased. In the brain, changes typical for adult copper metabolism, were not discovered. But in cerebrospinal fluid copper and Cp concentration sharply increased. The role of milk Cp in controlling the copper balance in newborn rats and the tissue-specific mechanism of Cp gene activity regulation are under discussion.

[In vivo expression of copper transporting proteins in rat brain sections].

Expression of two copper-transporting P1-type ATPases (ATP7A and ATP7B), the CTR1 protein, a high-affinity copper transporter, and ceruloplasmin (CP), a copper-containing ferroxidase. The level of mRNA of these proteins was determined by RT-PCR analysis, the distribution of polypeptides encoded by these genes was determined by immunoblotting, and the type of cells expressing these genes was identified immunohistochemically. It was found that the major product of CP gene in the brain is cell membrane-bound CP. Secretory CP, whose molecule contains the greatest number of weakly associated copper atoms, is synthesized in the vascular plexus. CTR1 mRNA is evenly distributed in the brain; however, its content is twice higher in the vascular plexus. The Atp7a gene is active in all brain sections, whereas the Atp7b gene is active only in the hypothalamus. The membrane-bound CP is expressed in glial cells of all types and in ependyma cells. ATP7b and ATP7a are expressed predominantly in ependyomyocytes and neutrons, respectively. The organization of copper transport in mammalian brain is discussed.

Isolation and partial characterization of cDNA clone of human ceruloplasmin receptor.

An individual clone, presumably carrying a 3 bp fragment of ceruloplasmin receptor cDNA was isolated from the expression library of human placenta cDNA using polyclonal specific antibodies to ceruloplasmin receptors. EcoR1-hydrolysate of isolated DNA was cloned in a pTZ19 bacterial vector and sequenced in the forward and reverse direction. The comparison of the revealed sequence with known sequences of human genome revealed its high similarity to ceruloplasmin cDNA.

[Further study of experimental hepatitis E in piglets]. / Dal'neishee izuchenie éksperimental'nogo gepatita E u porosiat.

Experimental infection of piglets with a fecal pool containing hepatitis E virus (HEV) strains Osh-25 and Osh-228 confirmed previous results obtained with HEV strain Osh-205 on susceptibility of these animals to hepatitis E. A biphasic increase of transaminase activities, histopathologic changes in the liver, virus excretion with feces were observed in the animals. The genome of viral particles in piglet feces conformed, at least partially, to human HEV RNA. Ultrastructural abnormalities of piglet hepatocytes resembled those in primates with experimental hepatitis E. The number of positive results (during follow-up of the particles and HEV RNA in various organs of experimental animals) was the highest on day 10 after infection. Both the particles and HEV RNA were present in the lymphoid tissue over the entire follow-up period, from day 5 to day 15 postinfection.

Identification of the linkage of mutations causing cystic fibrosis to different alleles of a tetranucleotide repeat in intron 6a of the CFTR gene.

The linkage of the intragenic polymorphic (GATT)n repeat to a number of cystic fibrosis transmembrane conductance regulator gene mutations (delta F-508, G542X, G551D, R553X, R1162X, W1282X, N1303K, R334W, and R347P) was studied. The linkage of delta F-508, G542X, and N1303K to a six-copy allele and of R334W to a seven-copy allele of the repeat was found.

Linkage disequilibrium between cystic fibrosis mutations and polymorphic 4-bp repeat within CFTR gene.

The PCR technique was used in a study of the linkage of cystic fibrosis mutations and a polymorphic (GATT)n repeat in intron 6 of the CFTR gene. Absolute linkage disequilibrium was found between the common delta F-508 mutation and the (GATT)6 allele. This allele was also in linkage disequilibrium with other unidentified mutations in the CFTR gene resulting in the pancreatic insufficient form of disease. The frequency of (GATT)n alleles in the pancreatic sufficient form of CF did not differ significantly from the data obtained in the total population. The significance of the (GATT)n polymorphic repeat for the diagnosis of CF is discussed.

[Allele polymorphism of the DNA loci MET, D7S8, D7S23, linked to the cystic fibrosis gene in some populations of the USSR, in high risk families and in cystic fibrosis patients]. / Allel'nyi polimorfizm DNK-lokusov MET, D7S8, D7S23, stseplennykh s genom mukovistsidoza, v nekotorykh populiatsiiakh SSSR, v sem'iakh bysokogo riska i u bol'nykh mukovistsidozom.

RELP analysis of DNA loci MET, D7S8 and D7S23 was carried out in Leningrad population and partially in populations of Moscow, Azerbaijan, Ukraine, Buryatia as well as in individuals from high risk families and in cystic fibrosis (CF) patients by means of blot hybridization and polymerase chain reaction. Allelic polymorphism of all loci studied in these three groups was found to be quite similar to that in the North-Western Europe and in whites of the North America. Linkage disequilibrium of the alleles studied with the CF gene was especially pronounced for alleles of the D7S23 locus and gradually decreases from KM-19 through CS-7 to XV-2c DNA probes. The data witness genetic homogeneity of the CF mutation in European populations of the USSR and its similarity to this mutation in Western Europe. The significance of these data for potential diagnosis of CF and for heterozygous carrier detection is discussed.

[Expression of ceruloplasmin gene in mammalian organs from the data of hybridization analysis with complementary DNA probes]. / Ekspressiia gena tseruloplazmina v organakh mlekopitaiushchikh po dannym gibridizatsionnogo analiza s komplementarnymi DNK-zondami.

Expression of ceruloplasmin (Cp)-coding gene in rat and human liver and brain tissues was studied by Northern blot hybridization and by in situ hybridization with cloned species-specific cDNA probes. In rat brain structures, different levels of Cp mRNA were detected, the maximal one was found in cerebellum. The steady-state level of Cp mRNA in rat and human brain was several times lower than in parenchymatous liver cells. The size heterogeneity of Cp mRNA was found. Polyadenylated RNA prepared from human liver contains two equally abundant Cp mRNAs differing in their chain length (3.6 and 4.5 kb) while brain polyadenylated RNA contains a single Cp mRNA (4.5 kb).

[Expression of ceruloplasmin gene in various organs of the rat]. / Ekspressiia gena tseruloplazmina v razlichnykh organakh krysy.

The contribution of different rat organs to the synthesis of ceruloplasmin (Cp) was studied. Dot hybridization with the use of the Cp cDNA probe revealed Cp mRNA sequences in RNA preparations from liver, heart, kidney as well as from different divisions of brain, the concentration of Cp mRNA sequences being maximal in the liver. Polyribosomes isolated from these organs effectively synthesized Cp in a cell-free system derived from rabbit reticulocytes. After in vivo pulse labeling, the newly formed radioactive Cp was detected in the membrane fractions from all these organs. The newly formed Cp was concentrated within the membranes of the Golgi complex of various organs where it was revealed by different immunochemical techniques. Experiments with isolation of the liver from the systemic circulation showed that the liver is the only organ secreting Cp into the blood stream. It was suggested that mammalian tissues contain at least two molecular forms of Cp, i. e., circulatory and intracellular ones.

[Polymorphism of nucleotide sequences of human genomic DNA linked to a mucoviscidosis locus]. / Polimorfizm nukleotidnykh posledovatel'nostei genomnoi DNK cheloveka, stseplennykh s lokusom mukovistsidoza.

Polymorphism in the restriction fragments length of human DNA sequences linked to mucoviscidosis locus was studied in the healthy control group and in the families affected by mucoviscidosis. The plasmid clones metH, pJ3.11,XV-2c and pKM.19 were used as hybridization probes. The allelic frequencies of the polymorphic loci were determined for total population and for affected families. The linkage disequilibrium between the disease locus and linked polymorphic loci detectable with XV-2c (TaqI endonuclease) and pKM.19 (PstI endonuclease) was demonstrated. The high informational value of DNA-diagnosis of mucoviscidosis in the family studies with the use of four DNA probes combination has been demonstrated.

[Comparative analysis of recombinant bacteriophages containing the sequences of the chromosomal ceruloplasmin gene in the rat]. / Sravnitel'nyi analiz rekombinantnykh bakteriofagov, soderzhashchikh posledovatel'nosti khromosomnogo gena tseruloplazmina krysy.

Three clones carrying the sequences of ceruloplasmin gene were isolated from the library of EcoRI fragments of rat chromosomal DNA cloned in Charon 4A vector. These clones were identified using, i) plaque hybridization with [32P] cDNA transcribed from highly purified rat ceruloplasmin (Cp) mRNA; ii) blot hybridization of the restriction fragments of recombinant DNAs with Cp cDNA and iii) hybridization selection of Cp mRNA followed by its cell-free translation. Oligo (dT)-primed cDNA transcripts of Cp mRNA having different length as well as cloned Cp cDNA isolated from rat liver cDNA library were used as hybridization probes for the study of mRNA-coding segments of Cp gene. The length of inserts in recombinant DNAs varied from 7.5 up to 12.3 megadaltons. EcoRI-fragments of Cp gene were mapped within recombinant DNA and their homology to each other was studied.

[Molecular mechanisms of the control of ceruloplasmin synthesis by estradiol]. / Molekuliarnye mekhanizmy regulirovaniia sinteza tseruloplazmina pod deistviem estadiola.

Mechanisms of the effect of estradiol on the biosynthesis of ceruloplasmin (CP) in rat liver were studied. The radioimmunological tests revealed that estradiol caused an increase of CP concentration in plasma. The about two-fold increase of plasma CP level was accompanied by the proportional increase of the content of CP nascent chains in membrane-bound polysomes of liver. Cell-free translation of free and membrane-bound polysomes revealed that there is a two-fold increase in the relative rate of CP synthesis by membrane-bound polysomes from estradiol-treated rats, while free polysomes did not contribute to CP synthesis. The similar increase of the rate of CP synthesis was found also when RNA preparations from membrane-bound polysomes and from post-polysomal supernatant were translated in a heterologous cell-free system. Studies on the kinetics of hybridization with a specific cDNA probe showed that the increase in the rate of CP synthesis correlates to the increase of the steady-state level of CP mRNA in both membrane-bound polysomes and post-polysomal supernatant. A conclusion is made that the induction of CP by estradiol is related to the accumulation of CP-mRNA in liver cells.