[Morphological aspects of diagnosis of acute opiate poisoning]. / Morfologicheskie aspekty diagnostiki ostrykh otravlenii opiatami.

Clinical anatomical analysis, forensic chemical, morphological, bacteriological, and immunological investigations of material from 62 subjects dead from acute poisoning with opiates before hospitalization (31 cases) and in hospital (31 cases) were carried out. Organ and tissue lesions typical of opiate poisoning were detected. The outcome of acute poisoning with opiates is largely determined by severe dystrophic and inflammatory processes in organs and tissues, typical of narcomaniacs.

[Hypophyseal damage in persons with cerebrocranial injuries and who died in the hospital]. / Povrezhdeniia gipofiza u lits s cerepno-mozgovoi travmoi, umershikh v statsionare.

The pituitary was examined in patients with craniocerebral injuries who died in hospital in various periods after treatment. Control group consisted of victims died at the site of accident. The results indicate the significance of examining the pituitary in craniocerebral injuries for the diagnosis of thanatogenesis, particularly in patients died in hospital. Causes of traumatic changes in the organ were determined, highly incident in the practice of forensic medical experts: directly during injury, as a result of skull bone fractures; during development of dislocation syndrome and in disorders of blood and lymph circulation in the brain matter; resultant from augmenting traumatic edema of the brain.

[Morphological changes in lungs of patients with spinal injuries died in hospital]. / Morfologicheskie izmeneniia v legkikh u lits s travmoi pozvonochnika, umershikh v statsionare.

Morphological characteristics of changes in the spine and lungs in patients with spinal injuries dead in hospital are described. Spinal edemas were detected in all cases. Edemas were found even in subjects dead at the place where the injury was inflicted. Morphological changes in the lungs were characterized by a phase-wise process, depended on the volume of injury, duration of hospitalization, medical care rendered, and manifested by disorders of blood content of the organ, development of tissue edema, and pneumonia. These data do not rule out the development of pneumonia in patients with spinal injuries.

[Morphologic changes in spinal cord of patients with spinal injuries who died in hospital]. / Morfologicheskie izmeneniia spinnogo mozga u lits s travmoi pozvonochnika, umershikh v statsionare.

Morphologic changes in the spine have been investigated in patients with spinal injuries who died in hospital. The incidence of injuries has been evaluated as 9.3% of mechanical injuries. Purposeful search for possible injuries increased the rate of detection of injuries of the spine and the cord. Spinal cord can be impaired without involving the osseous formations and spinal ligaments. Edema of the cord tissue develops during the early period of injury and prevents the detection of injuries, persisting for a long time after the injury. Edemas are detected even in subjects who died at the site of accident.

[Changes in antigenic properties of lipopolysaccharides of Shigella sonnei phase I having lost the virulence due to transposon integration into the invasiveness plasmid PSS120]. / Izmenenie antigennykh svoistv lipopolisakharida shigell Zonne I fazy, utrativshikh virulentnost' v sviazi s integratsiei transpozona v plazmidu invazivnosti PSS120.

A nonvirulent strain of Shigella sonnei phase I has been obtained by integration of the transposon Tn5 into the invasiveness plasmid pSS120 in the virulent strain and designated NR18. The presence of the plasmid pSS120 in both strains results in the similar morphology and bacterial ability to agglutinate in the presence of antiserum to Shigella sonnei phase I antigen. The lipopolysaccharide preparations from the virulent and nonvirulent strains give the similar reactions with the antiserum in the reaction of hemagglutination. However, in the reaction of passive local hemolysis in the gel (Jerne reaction) the significant difference is revealed in the immunogenicity of the preparations, with the preparations from the virulent strain being 4-5 fold more immunogenic. In crossreaction, the antibodies secreted by the mouse spleen cells immunized by LPS from the virulent strain show a weak reaction with the ram erythrocytes sensitized by the LPS of the nonvirulent strain. Thus, the biological changes in the LPS of the nonvirulent strains that are, evidently, the consequence of the structural changes, are identified only by the most sensitive immunological techniques.

[Plasmid pSE21 of Streptomyces erythraeus strains]. / Plazmida pSE21 shtamma Streptomyces erythraeus.

Crossing of S. erythraeus 4, a laboratory strain (NRRL 2338) containing a family of plasmids with S. erythraeus 1, a plasmid-free strain resulted in formation of strain 6. A multi-copy plasmid pSE21 11.5 kb in length was isolated from S. erythraeus 6. A detailed restriction map of plasmid pSE21 was constructed. Its cloning to E. coli YM83 on vector pUC19 showed that plasmid pSE21 was not stable in E. coli. It was found that the status of plasmid pSE21 changed in relation to the physiological state of S. erythraeus 6. Southern hybridization of the plasmid pSE21 DNA with the total DNA of the cultures of S. erythraeus 1 maintained for various periods at 4 degrees C demonstrated that plasmid pSE21 was present in S. erythraeus 1 in an autonomous state in 0.1 to 0.2 copies per genome. The number of the plasmid pSE21 copies could be decreased. The chromosomal DNA of S. erythraeus 1 contained the DNA sequences highly homologous to those of plasmid pSE21. It was assumed that during the crossing of S. erythraeus 4 with S. erythraeus 1 the genetic element from the donor strain was transferred to the recipient strain which in some way changed the plasmid pSE21 status and imparted the multicopy pattern to it. Further investigation of the plasmid pSE21 properties and construction of a vector for S. erythraeus on the plasmid basis are under way.

[Study of the effect of protoplast formation on the antibiotic activity of erythromycin producers]. / Izuchenie vliianiia protoplastirovaniia na antibioticheskuiu aktivnost' produtsenta éritromitsina.

The influence of protoplasting and regeneration on the strains of the erythromycin-producing organism differing by their origin was studied with respect to changes in the antibiotic production property. 223 regenerates of the erythromycin-producing culture were tested in several generations and it was shown that there was a marked change in the range of the variation by that property. 40 to 70 per cent of the variants in the IInd generation increased their levels of erythromycin biosynthesis by 20 to 60 per cent as compared to the intact cultures. However, in the subcultures the antibiotic production level decreased and by the IVth generation only 3 to 6 per cent of the variants preserved its increase by 10 to 20 per cent over the control level because of the culture high instability.

[A library of genes of plasmid PSS120 of Shigella sonnei]. / Biblioteka genov plazmidy PSS120 Shigella sonnei.

The gene library of S. sonnei plasmid pSS120 was constructed with the use of plasmid pSL5 as vector. The complete restriction of the vector DNA and the partial restriction of the DNA of plasmid pSS120 were carried out by means of the enzyme EcoRI. The restricted DNA was ligated and packed in vitro into the capsid of phage lambda. The titer of negative colonies obtained after packing was 0.8 X 10(3) clones per 1 microgram of S. sonnei DNA. The total number of detected clones was 250. On the basis of the results, obtained in the analysis of the inserts of the DNA of plasmid pSS120 into the DNA of recombinant clones, the theoretical volume of the library, equal to 92 clones, was calculated. The collection of clones thus obtained will be used for checking the presence of the determinants of invasiveness and phase I antigen, localized in the DNA of plasmid pSS120.

[Isolation and properties of the protoplasts of Streptomyces erythraeus]. / Poluchenie i svoistva protoplastov Streptomyces erythraeus.

The paper is concerned with development of conditions for cultivation of the erythromycin-producing organism and preparation and maintenance of its stable viable protoplasts. Optimal conditions for the culture growth and protoplasting were developed. Two-stage cultivation of the organism on media PB and S provided dense diffuse or diffuse local growth characterized by low differentiation and higher homogenicity. The incubation time at stage I was 66 hours and that at stage II was not more than 24 hours at respective temperatures. The culture was incubated on a shaker in the presence of glycine at the minimum concentration. The presence of glycine in the medium altered the culture cell walls which was evident from changing of staining by Gram from + to +/- . Treatment of such a culture with lysozyme for 30 minutes provided formation of up to 2.10(9) protoplasts per 1. ml. It is possible to maintain the protoplasts in frozen state at -20 degrees C in medium P for 1 month. Under such storage conditions the titer of the viable protoplasts as compared to the initial one decreased only 2-fold after the one-month storage.

[Identification and characteristics of plasmids of the strains of erythromycin-producing Streptomyces erythreus]. / Identifikatsiia i kharakteristika plazmid v shtammakh Streptomyces erythreus-produtsentakh éritromitsina.

Presence of plasmid DNA was investigated in laboratory strains 2 and 4 (NRRL 2338) of S. erythreus, as well as in strains 1 and 3 of S. erythreus subjected to improvement with respect to erythromycin production. Families of plasmids close by their molecular weights were identified in S. erythreus strains 3 and 4 (NRRL 2338). A plasmid DNA fraction of S. erythreus strain 3 was studied with electron microscopy. It enabled to identify 5 plasmids: pSE11, pSE12, pSE13, pSE14 and pSE15 with length of 5.3, 12.4, 16.3, 29.6 and 86.9 kb respectively. Using of various procedures for isolation of extrachromosomal DNA did not provide its detection in S. erythreus strains 1 and 2. At least a part of the plasmids detected in S. erythreus strains 3 and 4 (NRRL 2338) was conjugative. 32R-Labeled plasmid DNA of S. erythreus strain 3 was subjected to hydridization according to Sauthern with total DNA of the 4 strains treated with restrictases BamHI, PstI and BgIII. The studies showed that the genome of S. erythreus strain 2 was not homologous with the probe while S. erythreus strain 1 contained one of the plasmids or its part in chromosome-integrated state. In strains 3 and 4 (NRRL 2338) of S. erythreus certain plasmid DNAs were present in both autonomous and chromosome-inserted states. 32P-Labeled gene of erythromycin resistance (ermE) was subjected to hybridization according to Southern with total DNA of the 4 strains and with DNA plasmid fraction of S. erythreus strain 3. The signal was positive only in hydridization of the probe with total DNA of S. erythreus strains 1, 3, and 4 (NRRL 2338).(ABSTRACT TRUNCATED AT 250 WORDS)

[Cloning of the purA16 locus in Rec+ cells of Bacillus subtilis]. / Klonirovanie lokusa purA16 v Rec+-kletkakh Bacillus subtilis.

A portion of purA16 chromosomal locus of Bacillus subtilis was cloned into Rec+ cells of this microorganism with pBD12 plasmid (carrying chloramphenicol and kanamycin resistance determinants) serving as a vector. The hybrid plasmids were stably maintained in cells grown on media supplemented with antibiotics and were lost from cells in the absence of drugs. The cloned fragment could incorporate into the chromosome some with a frequency of 10(-2) per cell per generation. A clone carrying the hybrid plasmid inserted into the chromosome was detected.

[Transformation and transduction in Bacillus subtilis strains with the BsuR restriction-modification system by modified and unmodified pUB110 plasmid DNA]. / Transformatsiia i transduktsiia u shtammov Bacillus subtilis s sistemoi restriktsii-modifikatsii BsuR posredstvom modifitsirovannoi nemodifitsirovannoi DNK plazmidy pUB110.

Plasmid pUB110 DNA is restricted and modified during transformation of Bacillus subtilis cells possessing the BsuR system of restriction-modification. Restriction has comparatively little influence on the frequency of plasmid transformation only reducing it 20 times. The frequency of transduction of nonmodified plasmid DNA into modifying recipient cells, using phage Ar9 is also reduced a little. The frequency of transduction of chromosomal markers is much more lowered under the same experimental conditions.

Transformation and transduction of Bacillus subtilis strains with the Bsu R restriction-modification system by means of modified and unmodified DNA of pUB110 plasmid.

During transformation of B. subtilis cells with the Bsu R restriction-modification system by means of pUB110 plasmid restriction and modification of the plasmid DNA occurs. The effect of restriction on the transformation frequency is relatively weak, bringing about 20-fold decrease only. When using cells of a modifying recipient, the frequency of AR9 phage-mediated transduction of unmodified plasmid DNA is also relatively little decreased. The frequency of transduction by chromosomal markers, under the same conditions, falls much lower.