[Apoptosis and neutrophils on the regulation of proliferation and differentiation ex vifo of myeloid cells with Ph-chromosome].

Human myeloid cells with Ph chromosome (Ph+ cells) from chronic myeloid leukemia (CML) in the course of proliferation and differentiation ex vivo are regulated under alternation of cell proliferation and neutrophil maturation stages by consecutive blocking and inducing apoptosis with of neutrophils participation as well bcr/abl, bax and bcl2 genes expression. Apoptosis regulation of three main Ph+ cells types from CML patients depends on alternation sequences of proliferation (1) and maturation (2) cell stages and realized by two ways. The first one is performed by consecutive blocking and inducing apoptosis under 2/1/2 stage alternation. The way is not described early. Neutrophils accumulation correlates with apoprosis blocking. Apoptosis level enhances under neutrophils exhausted. Apoptosis blockage allows cells to proliferate and, thus, to form new portion of neutrophils with consecutive regular their death as well a consequent alternation of apoptosis blocking and inducing. This way regulates proliferation efficiency indexes P/D that reflect Ph+ cells proliferating potential and performs cycle completion for proliferation and differentiation. The second way of apoptosis regulation starts from proliferation stage and performs for 1/2/1 alternations under diminished content of neutrophils and a little increase under next maturation. It leads to resistant depressed apoptosis levels that, at maximal points, are 3-8 times lower than those under alternation 2/1/2. Resistant apoptosis blocking is observed in the Ph+ cells with prolong proliferation or maturation stages, when blasts and myelosytes are accumulated under enhanced bcr/abl and bcl2 > box gene expression and remain under next maturation. Stable apoptosis blocking is accompanied by increasing amounts of blasts and myelocytes and enhancing bcr/abl and bcl 2 > bax expression. This is observed under CML progression. Ph+ cells cultivation may be useful for more distinct diagnostics of CML phases of individual CML patients and optimization of the treatment.

A pea mutant (costata) expressing higher activity in thylakoid membrane-bound carbonic anhydrase alters PSII downregulation mechanisms.

The interaction between photosynthetic electron transport and the activities of the thylakoid associated carbonic anhydrase (tCA), estimated as combined tCA activity in pea plants (Pisum sativum L. Borek cv., WT) and mutant form (costata 2/125) that differ in chlorophyll content have been compared. Chlorophyll a fluorescence changes after the inhibition of tCA by ethoxyzolamide (EZ), estimating possible role of tCA in PSII downregulation were investigated. Costata expresses higher tCA activity and higher O2 evolution in comparison to WT. Inhibition of tCA by EZ decreased effective PSII photochemistry that coincided with an enhancement in thermal dissipation, while maximal PSII quantum yield (F(v)/F(m)) did not significantly change. Ethoxyzolamide induced changes in fluorescence parameters that were more strongly expressed in costata 2/125. The results show that tCA is involved in the regulation of the proton gradient across thylakoid membranes and thus limits PSII downregulation.

[Kinetics of proliferation, differentiation and transcription of genes regulating apoptosis in cultured human BCR/ABL+ Ph+-cells].

Ph+, bcr/abl+ cells arise due to t(9,22) chromosome translocation and Ph+ chromosome formation in hematopoietic stem cells. The cells show appreciable apoptosis suppression but retain their ability to differentiate and maturate. Ph chromosome, bcr/abl oncogene and Ph+, bcr/abl+ cells themselves are the hallmark of chronic myeloid leukemia. Under leukemia progression differentiating Ph+, bcr/abl+ cells transform into leukemic malignant cells with differentiation block. It is assumed to be a result of subsequent mutations or activation of proliferation of long silent Ph+ cells arisen previously in the stem cells because of the translocation. Real mechanism underlying the cell transformation remains unknown. This work was performed to develop a proper cell model allowing us to study functioning of differentiating Ph+, bcr/abl+ cells and their real transformation into malignant cells with block of differentiation. For this purpose we have investigated kinetics of Ph+, bcr/abl+ cells proliferation, differentiation, cell death and transcription of antiapoptotic genes in cultured 14-day of Ph+ mononuclear cells isolated from peripheral blood of a patient in chronic phase of chronic myeloid leukemia before treatment. The results obtained revealed that Ph+ cell differentiation proceeded in accord with characteristic scheme of chronic myeloid leukemia in vivo. Myeloid cells of hematopoietic cell lineage amounted to 3/4 of live Ph+ mononuclear cells undergoing accumulation and subsequent consumption in the course of differentiation. 95% myeloid cells were differentiating Ph+ granulocytes. The most deal of differentiating Ph+ cells was myelocytes. The rate ratio of myelocyte accumulation to its subsequent consumption showed that the rate of transformation into metamyelocytes was significantly decreased at this differentiation stage. Ph+ cells cultivation curves characterized cell death at different differentiation stages. There were observed the cell death of proliferating Ph+ cells and Ph+ myelocytes, and intensive death of mature cells as well. P/D index, that is ratio of immature Ph+ granulocytes differentiated by cell dividing (blasts, promyelocytes and myelocytes) to the cells differentiated without dividing (metamyelocytes and mature neutrofiles), revealed active of proliferation at the beginning of cultivation and unexpected new proliferative activity at the end of cultivation in the presence of growth factor. The peaks of antiapoptotic bcr/abl gene transcription activity coincided with the observed active proliferation at the beginning and at the end of cultivation. Cell proliferation, differentiation and apoptosis were noticeably accelerated by growth factor treatment. Thus, the study of the Ph+ cells cultivation kinetics is rather informative approach to investigation of continuous regulation of cellular and molecular processes in vitro in the case of chronic myeloid leukemia and allows more complete consideration of Ph+, bcr/abl+ cells hematopoiesis.

[Surgical operations in women over 60 years of age in the gynecological practice (according to the data from Obstetric and Gynecology Clinic of Stara Zagora during 1998-2002 years)]. / Operativni interventsii v ginekologichnata praktika pri zheni nad 60 godini (po materiali na AG klinika Stara Zagora za perioda 1998-2002 god).

In period of 1998 to 2002 year in Klinik of Obstetric and Gynecology Stara Zagora was performed 2161 small and 1122 great gynecologic operations in women older 60 years. The most patients was in age of 60 to 70 years (79%), 17.9% was in age of 70 to 80 years and over 80 years was 2.9%. The neoplasm disorders perform 34.6% and static disorders--29.6%. By 84.5% in women with postmenopausal bleeding no established neoplasm and by 15.5% was established endometrial cancer. The great operations included 55.6% abdominal hysterotomy, 13.6% vaginal hysterectomy and 9.8% urogynecologic operations. Postoperative complications perform 3.7% (2 patients).

Effect of carnosine on immunocompetent cells from alcoholic patients.

We studied the effect of natural dipeptide carnosine on phagocytosis, respiratory burst in neutrophils, and subpopulation composition of lymphocytes from healthy donors and alcoholic patients. Carnosine in vitro produced different effects on immunocompetent cells from healthy donors and patients with alcoholism. In patients with alcoholism phagocytic activity of leukocytes and generation of reaction oxygen species increased under the influence of carnosine in a concentration of 0.01 mM, but decreased after treatment with this compound in a concentration of 1 mM. Carnosine in both concentrations stimulated the respiratory burst, but had no effect on the count of phagocytic cells in healthy donors. Carnosine in a concentration of 0.01 mM increased the number of lymphocytes carrying apoptosis markers (CD95+) in patients with alcoholism not receiving therapy. Our results indicate that carnosine holds much promise for the therapy of alcoholism.

[The characteristics of the interferon system parameters in typologically different groups of patients with borderline mental disorders during treatment]. / Kharakteristika nekotorykh parametrov sistemy interferona v tipologicheski razlichnykh gruppakh bol'nykh s pogranichnymi psikhicheskimi rasstroistvami v protsesse lecheniia.

Lymphocyte gamma-interferon production and levels of serum interferon were studied in 94 patients with borderline psychiatric disorders on treatment. It was shown that parameters of interferon system in the acute disease period and their dynamics through the treatment depend on typological features of personality and initial level of interferon production. The patients with aggressive features of personality in acute disease exhibit significantly higher levels of serum interferon than those in patients with submissive features which drop after recovery. The same tendency occurred in hysterical patients (hystero-expressive and hystero-impressive types). These findings may be due to different reaction of immune system to chronic emotional stress in different types of personality.

[Cytogenetic instability, the ability to produce interferon and immunoreactivity of blood lymphocytes in patients with schizophrenia]. / Tsitogeneticheskaia nestabil'nost', sposobnost' produtsirovat' interferon i immunoreaktivnost' limfotsitov krovi bol'nikh shizofreniei.

In schizophrenia patients, the level of T-lymphocytes with cytogenetic aberrations is elevated, and the ability of these cells for PGA-blast-transformation and the interferon synthesis is reduced. The elevation of the lymphocytes ability for blast-transformation in the presence of the native preparation of DNA and for lysis of the erythrocytes proper is marked in these patients. The observed changes were especially marked in schizophrenia with an interrupted course.

[The interferon system of patients with schizophrenia during therapy]. / Sistema interferona u bol'nykh shizofreniei v protsesse terapii.

The status of the interferon system was studied in 100 schizophrenics, in 40 of them over time (during exacerbation of the process and after psychotropic therapy). It has been established that the lymphocyte production of alpha- and gamma-interferon is depressed during exacerbation of the disease. Following treatment lymphocyte production of alpha-interferon remained low, while the production of gamma-interferon normalized.

[Multicausality in the pathology of commercial swine breeding]. / Multikauzalnost v patologiiata na promishlenoto svinevudstvo.

Studied were more than 100,000 samples of pathologic tissue material, 1,900 whole blood and blood serum samples, 8,000 fecal samples, 2,600 washings samples, swab samples, and other sampled material via the employment of a number of bacteriologic, virologic, morphologic, hematologic, biochemical, parasitologic, and other routinely applicable laboratory methods over a period of 5 years. All materials were taken from 15 stock swine breeding complexes, 3 reproduction and 2 nucleus bases for hybrid pigs, and 3 swine breeding farms with a practice of extensive raising technology. It was demonstrated that the adaptive capacity of the animals of the Camborough hybrid sharply dropped when the pigs were kept under unconventional conditions of raising, leading to deviations in the application of industrial technology methods. This was the reason to observe the simultaneous manifestation of conditionally substantiated disease outbreaks (multiple causality), with great economic losses. Studied were the etiology and pathogenesis of coli bacteriosis, salmonellosis, gastric ulcer, hypotrophy, atrophic rhinitis, geohelminthiases, chlamydial infections, and erysipelas. Suggested is the use of programmes and methods of prophylaxis and treatment, of expedient diagnosis and control on immunity.

[Associated vaccination of pigs against swine fever, erysipelas and Aujeszky's disease]. / Asotsiirano vaksinirane na praseta sreshtu chuma, cervenka i bolestta na Aueski.

Tested was the simultaneous vaccination of pigs against swine fever, swine erysipelas, and Aujeszky's disease. Parallel to this experiment a second group of test pigs were treated with each of the three vaccines at twenty-day intervals. The effect of vaccination was assessed through clinical observations with the test groups and by challenging part of the animals with virulent strains of the swine fever and erysipelas agents. It was found that the successive application of the three vaccines at 20-day intervals (at the age of 50, 70, and 90 days) could induce immunity that lasted for a longer period than the immunity resulting from the simultaneous treatment at the age of 90 days. In order to protect the pigs for sure up to the end of the fattening period a revaccination against swine fever with the simultaneous use of the vaccine against swine erysipelas in both cases was necessary.

[Pathology in hybrid pig breeding due to mixed infections]. / Patologiia na khibridnoto svinevudstvo, prichineno ot smeseni infektsii.

A total of 7,500 samples from 5 swine breeding complexes with 200,000 hybrid Camborough pigs were studied morphologically and bacteriologically. The raising technologies used on all complexes were of industrial type or semi-industrial and extensive. A description is given of the simultaneous participation of the following nosologic units: colibacteriosis, responsible for 13 to 14 per cent of the total mortality rate in newborn pigs; bronchopneumonia--causing 6 to 39 per cent losses in the other age groups; and dysentery with salmonellosis--inflicting 5 to 9 per cent losses. Underlined is the great advantage of the industrial technologies of animal raising as against the conventional ones in the prevention of these diseases in hybrid pig breeding. Tested were effective programmes for the prophylaxis associated with the individual nosologic units. More essential were the following: active immunoprophylaxis with live attenuated vaccines against colibacteriosis and salmonellosis; elimination of the alimentary stress factor in dysentery; disinfections of premises by gassing with a mixture of ammonium nitrate and chlorinated lime in a 1:2 ratio. However, all measures proved effective when the principle of "all in, all out' was observed which is essential part of the Camborough industrial raising technologies.

[Experiments to produce an attenuated strain of the transmissible gastroenteritis virus and its use as a live vaccine]. / Opiti za polchavane na atenuiran shtam na virusa na transmisivniia gastroenterit i izpolzuvaneto mu kato zhiva vaksina.

Serial passages have been performed of a field virus of the transmissive gastroenteritis to obtain an attenuated strain adapted to a permanent cell line of pig kidney, "SPEV". The strain is innocuous for test animals, for pigs of all ages, and for swine during the entire period of pregnancy. It has proved genetically stable. The strain is immunogenic for swine and pigs both under laboratory and under field conditions (in new foci and on stationary farms) and can be used as a live vaccine against transmissive gastroenteritis, either untreated or in a freeze-dried state.

[Principles for evaluating reactivity in the clinical picture of alcoholism and addiction during drug therapy]. / O printsipe otsenki reaktivnosti v klinike alkogolizma i toksikomanii v khode medikamentoznoi terapii.

Eighteen patients with chronic alcoholism, stages II-III were examined during tisercin treatment and medicamental interval. In patients with stage II alcoholism, perversion of the central neuronal tisercin effect was shown. Phenothiazines efficacy during abstinence period appeared to be linked with the perversion observed. Individual constant control of blood pressure, heart rate, acetylcholinesterase serum level, sleep function etc. during drug treatment is considered to be a perspective trend for an increase of toxicomania treatment efficacy.

[Circular dichroism study of the reaction between the protein product of bacteriophage f1 gene 5 and synthetic polynucleotides]. / Izuchenie vzaimodeistviia belka--produkta gena 5 bakteriofaga f1 s sinteticheskimi polinukleotidami metodom krugovogo dikhroizma.

The complexes of gene 5 protein (phage f1) with single-stranded and double-stranded polynucleotides were investigated by circular dichroism (CD). In our experiments the concentration of the protein varied accordingly to polynucleotide. A decrease of the CD amplitude for polynucleotide-protein complexes was shown in all the regions of wavelength studied. The data indicate that the protein is bounded to the polynucleotide. This protein bounds differently to double-stranded polynucleotides containing ribo-ribo, ribo-deoxyribo and deoxyribo-deoxyribo chains. This difference is explained by differences in the form of the secondary structure of polynucleotides, containing ribo-ribo, ribo-deoxyribo and deoxyribo-deoxyribo chains.

Problems of bovine leucosis control and prophylaxis in industrial breeding conditions.

Industrial cattle-breeding in Bulgaria is characterized by large cattle-breeding complexes holding 500, 1,000 or 2,000 dairy cows; farms specialized in rearing calves for breeding purposes, and calves fattening units holding 2,000, 5,000 or 10,000 animals. A specific technology has been introduced for management purposes. This high concentration and specialization creates specific organisational problems for antiepizootic control measures. Bovine leucosis has been brought from abroad by imported breedings animals. A complex system for leucosis prophylaxis and control is obligatorily implemented. It is supervized and financed by the state. Particular care is taken of ensuring protective measures for import breeding animals. The infected farms are put under strict quarantine. The animals which show a positive reaction during complex diagnostic tests are slaughtered and the meat is used after heat-processing. The milk is heat-processed too and sent for further utilization. Calves born in infected farms are not fit for breeding purposes but for fattening only.

[Effect of the protein product of phage f1, gene 5, on heat denaturation of synthetic polynucleotides]. / Vliianie belka--produkta gena 5 faga f1--na teplovuiu denaturatsiiu sinteticheskikh polinukleotidov.

The influence of phage f1 gene 5 protein on melting of the synthetic polynucleotides has been investigated, using UV-spectroscopy. In our experiments we have varied the proteins concentration. It has been shown, that the protein lowers the melting temperature of the studied polynucleotides (d/A--Tn dAndTn, rAndTn, rAn.r n, dAn.rn). The melting temperatures and the shapes of melting curves of various polynucleotides differ when the same protein concentrations are used. We have shown that the protein binds to the double-stranded polynucleotides, containing ribo-ribo-, deoxyribo-ribo-chains. The difference in melting temperatures and shapes of melting curves was explained using the data about the differences in the secondary structure of these polynucleotides. Only for d/A-Tn renaturation was observed after sample cooling. It may reflect the single-stranded hairpin structure of this polynucleotide.

[Influence of protein bound with single-stranded DNA on the synthesis of RNA and poly(A). III. Protein product of F1 gene 5]. / Vliianie na sintez RNK i poli (A) belkov, sviazyvaiushchikhsia s odnonitchatoi DNK. III. Belok--produkt gena 5 faga fl.

Influence of the protein product of F1 phage gene 5 (protein 5) on the synthesis RNA and poly(A) in vitro was studied. It has been shown, that protein 5 has no effect on the transcription of the native DNA by E. coli RNA-polymerase, but completely prevents RNA and poly(A) synthesis on the denatured or single-stranded DNA at the protein/DNA ratio 10:1. Protein 5 inhibits poly(A) synthesis with oligo(dT)9 and oligo(dT)12 as a template, preventing binding of enzyme to the oligonucleotide. After the initiation of the poly(A) synthesis the inhibition becomes considerably wearer. The biological function of the inhibition of the transcription by "unwinding" proteins is discussed.

[Epizootiology of foot rot in sheep]. / Prouchvane vurkhu epizootologiiata na kopitniia gnilets po ovtsete.

Investigations were carried out on the outbreak and spread of foot rot in sheep as well as on the ways of its restriction and eradication. It was found that the disease had been introduced with the import of breeding animals, its further spread being associated with the activity of the Breeding Centers for pedigree animals. The diseased animals have been 25 per cent, on an average, for the whole country, and the economic losses, as expressed in production parameters, have amounted to: produce of animal origin--20 per cent; culling of animals--15 per cent; mortality--1 per cent. Best in the group treatment of the affected animals prove: the dipping in a 5--10 per cent solution of formalin or a 20 per cent solution of copper sulfate, combined with the respective surgical and orthopedic handling and the disinfection of premises. The individual parenteral treatment and the local application of antibiotics and chemotherapeutics have proved suitable and successful in the cases of moderate spread of the disease. The eradication of foot rot can be achieved only via the planned and complex application of pertinent measures.