Natural foci of ornithosis in Kazakhstan]

An investigation (clinical, immunological and epidemiological) of the workers of various fowl-farms was carried out in various regions of Kazakhstan for a number for years; the sera of domestic fowl and birds was studied as well. On the basis of examination of 6264 persons the greatest percentage of ornithosis was revealed among the workers of the slaughter house and the shop for the processing of the fowl. Examination of 4749 sera of bird blood showed that the highest incidence of ornithosis was among doves and ducks (43.2 and 11.6%), and the least--in chickens (5.3%); there proved to be a direct relationship between the infection of man and birds. Thus, it was shown that domestic fowl and also doves served as the source of ornithosis infection; foci of ornithosis were revealed in a number of Kazakhstan regions as a result of these investigations.     

Prevalence of ornithosis in the Lithuanian SSR

As a result of immunological examination of 658 domestic ducks of 6 poultry farms and 269 doves caught in Vilnius, antiornithosis antibodies were revealed in 3.1 +/- 0.7% of ducks and in 36.4 +/- 2.9% of doves. They were also found in 20 of 34 wild waterfowl. The causative agent of ornithosis was isolated in virological examination of the organs of a coot. Ornithosis nature of an epizootic outbreak among ducks of a poultry farm was proved by virological examination. The population was examined immunologically (2285 persons) and by intradermal test with the ornithosis antigen (80 persons). Positive specific reactions were revealed in 32.6% of persons engaged in poultry tending, and in 7.6% of other rural population. A case of ornithosis in man engaged in fowling (of doves) for their examination is described.     

Prevalence of ornithosis and other Chlamydia-like infections in the Ukrainian SSR

The prevalence of ornithosis and other Chlamydia infections among various groups of the population, some species of birds and mammals was studied in several regions of the Ukrainian SSR. Among the town population having no occupational contact with domestic fowl and farm animals, complement-fixing antibodies to group ornithosis antigen were found in 6.5% of the examined, among the inhabitants of rural localities in 8.4% and among the workers of poultry farms in 19.9%. During the examination of the immunological state of the workers of cattle-breeding farms, antibodies were detected in 15.1%. Pigeons, hens and ducks were investigated as probable sources of infection. The number of positively reacting individuals was 53.7, 13,7 and 24.9%, respectively. This indicates that they play a role in the spread of infection. The obtained data point out a considerable spread of Chlamydia infections among farm animals. Positive results of serological examination were recorded in pigs in 25.7%, in calves in 15.1% and in 8.6% of cases examined.     

Use of the indirect hemagglutination reaction in studying ornithosis infection. III. The diagnostic value of the IHR with an ornithosis erythocytic diagnosticum]

In both experimental and clinical conditions the passive hemagglutination test (PHAT with the use of an ornithosis erythrocytic diagnostic preparation was found to be sufficiently sensitive and specific as compared with the complement fixation test (CFT), a routine testing method. The study of the dynamics of immune response in infected animals and ornithosis patients allowed to regard the PHAT as a comparatively early method of serological analysis. Hemagglutinins were also found to circulate in the patients' blood sera only for a short time (on the average for 1 1/2--2 months). The CFT and the PHAT with erythrocytic diagnostic preparation, used in combination, will make it possible not only to diagnose ornithosis in patient more effectively, but also to differentiate between the cases of infection and anamnestic reaction.20     

Use of a cryoultramicrotomy method for the purpose of studying chlamydial ultrastructure]

The method of cryoultramicrotomy was adapted for the study of the ultrastructure of HeLa and McCoy cells in monolayer cultures infected with Chlamydia, obligatory intracellular procaryotic parasites, the causative agents of ornithosis (strain Loth) and paratrachoma (strain LB 1). The cryosections were obtained by the fixation of the monolayer with 2.5% glutaraldehyde, by the gradual infiltration of precipitated cells with sucrose (0.6--1.2--1.8--2.3 M) prior to freezing in liquid nitrogen, and by the treatment of sections with 1% aqueous methyl cellulose solution before drying. This method ensured good preservation of both Chlamydia, in intracytoplasmic inclusions and host cells, as well as regular reproducibility of the results. Ultrathin sections showed a considerable polymorphism in the vegetative forms of Chlamydia, which was probably due to the structure of their cell walls. Chlamydia, were found to form small vesicle-like structures in the cavities of inclusions. The cell walls and granules inside the elementary bodies of the causative agent of ornithosis were stained with the use of phosphotungstic acid--HCl, pH 0.5.     

Comparison of the ultrastructure of several rickettsiae, ornithosis virus, and Mycoplasma in tissue culture

Anderson, Douglas R. (National Cancer Institute, Bethesda, Md.), Hope E. Hopps, Michael F. Barile, and Barbara C. Bernheim. Comparison of the ultrastructure of several rickettsiae, ornithosis virus, and Mycoplasma in tissue culture. J. Bacteriol. 90:1387-1404. 1965.-In an effort to make a valid comparison of the ultrastructure of several intracellular parasites, selected agents were propagated under identical conditions in a single type of tissue culture cell; such infected preparations were processed for examination by electron microscopy by use of a standardized procedure for fixation and embedding. The organisms studied were: the Breinl and E strains of epidemic typhus, Rickettsia prowazeki; the Bitterroot strain of R. rickettsii; the Karp strain of R. tsutsugamushi (R. orientalis); R. sennetsu; the P-4 strain of ornithosis virus; and the HEp-2 strain of Mycoplasma hominis type I. Each of the rickettsial species examined had a cell wall and a plasma membrane, and contained ribosomes and deoxyribonucleic acid (DNA) in a ground substance. However, certain differences were noted. Both strains of R. prowazeki contained numerous intracytoplasmic electron-lucent spherical structures (4 to 10 mmu), not previously described. R. sennetsu, unlike the other rickettsiae, was not free in the host cytoplasm but was always enclosed in a vacuole. R. rickettsii was observed intranuclearly and in digestive organelles of the host cell as well as in the cytoplasm. Cells infected with ornithosis virus contained several forms representing the stages in its life cycle. The "initial bodies," made up of ribosomes and DNA strands, were morphologically similar to the rickettsiae. In cultures infected with M. hominis, most of the cells became large and multinucleate. Although the Mycoplasma organisms were readily cultivated from these cultures, only a few could be found in the electron microscope preparations. These organisms were extracellular and lacked a cell wall, being bound only by a unit membrane. Again, the internal components were ribosomes and DNA strands. Under the uniform preparative conditions employed here, the three groups of organisms were morphologically distinguishable from one another.     

The mechanisms maintaining epidemic foci of leptospirosis]

24,419 samples of blood serum, collected in the Azerbaijan SSR among different professional groups of the population, have been tested for leptospirosis. The comparison of the results with those obtained in the natural foci of leptospirosis and in the foci of this infection appearing as a consequence of human activities has made it possible to come to the conclusion that the epidemic foci of leptospirosis are maintained due to the existence of foci among cattle. The cases of contamination among persons professionally linked with large cattle breeding complexes have been found to exceed those among persons working at farms of a nonindustrial type.     

Results of the study of bird droppings collected in the Lithuanian SSR for the purpose of searching out tularemia epizootics

There are practically no records of cases of tularemia among humans in the Lithuanian SSR. Nevertheless, the mass sero-allergic survey of the population for tularemia, carried out 10-12 years ago, showed that 2.3% of the adult population in the Republic had had contacts with the causative agent of this infection. The work was aimed at the determination of the present activity of the foci of tularemia. During 6 years in 22 rural districts 2582 samples of avian excrements, containing bones and wool of small animals, were collected and studied by means of the antibody neutralization test (ANT). In 132 (5.1 +/- 0.4%) excrement samples collected on the territory of 12 districts Francisella tularensis antigen was detected. The average ANT titer was 45.2, the maximum titer (10 excrement samples) reached 1: 160. The study revealed the existence of the natural foci of tularemia in Lithuania at present, but their activity proved to be low. The most unfavorable situation was found to exist in western districts of the Republic.     

Changes in the phosphorylation and distribution of vinculin during nerve growth factor induced neurite outgrowth

The mechanism of neurite initiation and elongation was studied using nerve growth factor (NGF) treatment of PC12 cells. The distribution of focal adhesion sites and of the cytoskeletal protein vinculin was determined in large, fused, multinucleated PC12 cells. In the absence of NGF, focal adhesion sites as seen by interference reflection microscopy were restricted to the cell periphery in a regular distribution. Vinculin assemblies (foci), observed by indirect immunofluorescence microscopy using affinity purified anti-vinculin antibodies, were restricted to the cell periphery at focal adhesion sites. Within 4 hr after NGF treatment of the cells, the distribution of both vinculin and focal adhesion sites began to change. Focal adhesion sites became restricted to discrete protruding portions of the cell periphery. Larger, brighter vinculin foci appeared at the tips of the cell margin extensions, concomitant with the loss of foci at locations between the protrusions. As neurites elongated focal adhesion sites and vinculin foci remained with the tips of the growth cone extensions. Both focal adhesion sites and vinculin foci were rarely seen in the perikarya of cells with elongating neurites, and these were always confined to extended portions of the cell body margin. Occasionally, vinculin foci could be seen at the proximal portion of the neurite, at bending elbows, and at discrete expansions along the length. By immunoprecipitation of vinculin from 32P-labeled cells, vinculin phosphorylation was found to be increased within 1 hr of NGF treatment. The role of vinculin phosphorylation and assembly in the formation and directional elongation of neuritic processes in response to NGF is discussed.     

Coxsackie A 13 virus in the foci of rheumatism.

Faeces and nasopharyngeal washing were examined in 12 patients and 27 contacts of these patients in family foci (primary foci) of rheumatism and in 37 patients and 32 contacts of these patients in secondary foci of rheumatism (in hospitals). Results of stimultaneous examinations of 127 children of the control groups served as the control. An analysis of the results of the investigation showed the Coxsackie A 13 virus was found considerably more frequently in both the patients (83.3%) and their contacts (148.1%) in the primary foci of rheumatism than outside these foci (11.9--15.3%). The difference is statistically significant. Wide distribution of Coxsackie A 13 virus was also observed in clinical departments in which children were hospitalized at the acute stage of rheumatism. We failed to establish marked differences in the rate of detection of other viruses in the foci of rheumatism in comparison with the control groups.     

Tick-borne encephalitis in the Maritime Territory

On the basis of epidemiological analysis carried out at the period of 1940-1987, a decrease in tick-borne encephalitis (TBE) morbidity was registered; at the same time time the disease invariably took, as before, a clinically severe course. The most dangerous foci of TBE were found to be located in the southern Okhotsk region grown with dark coniferous forests. The subsiding and activation of the natural foci of TBE in different regions of the territory were established and some heretofore unknown foci in southern regions of the Maritime Territory were found. Ixodes mites inhabiting the Maritime Territory were shown to have a low level of virus carriership, thus causing the low level of population immunity to TBE virus. Combined foci of TBE and Powassan encephalitis were found.     

Use of the immunofluorescent microagglutination reaction for the purpose of studying ornithosis infection. 1. Production and approbation of a corpuscular luminescent diagnosticum]

A method of obtaining the fluorescent ornithosis corpuscular diagnostic agent providing for the extraction of the corpuscular antigen with its subsequent conjugation with fluorochrome-fluoresceine isothiocyanate. The use of this preparation permits to stage the agglutination reaction on the basis of immunofluorescent analysis, this facilitating the recording of the reaction results and considerably decreasing consumption of the diagnostic agent. As shown, the suggested immunofluorescent microagglutination reaction was characterised by a sufficiently high sensitivity, specificity, and can be used for detection of antibodies to the causative agent of ornithosis.     

Zoonotic infections in persons working in plants producing meat and bone meal

The present study has revealed that persons working at offal and bone processing plants show no specific pathology, peculiar to such plants, in the state of their health and severe mass diseases among these persons are absent. Nevertheless, rather frequent occurrence of erysipeloid should be noted. The survey of workers employed at different stages of the production process has revealed the presence of ornithosis, listeriosis and leptospirosis infections. No clinically manifest cases of these diseases have been detected. The infection of individual workers occurs in the process of production as the result of contacts with infected raw materials of animal origin. For preventing the possible spread of infection great attention should be paid to sanitary, hygienic and epidemiological measures.     

Plasmodium falciparum Sec24 marks transitional ER that exports a model cargo via a diacidic motif

Exit from the endoplasmic reticulum (ER) often occurs at distinct sites of vesicle formation known as transitional ER (tER) that are enriched for COPII vesicle coat proteins. We have characterized the organization of ER export in the malaria parasite, Plasmodium falciparum , by examining the localization of two components of the COPII machinery, PfSec12 and PfSec24a. PfSec12 was found throughout the ER, whereas the COPII cargo adaptor, PfSec24a, was concentrated at distinct foci that likely correspond to tER sites. These foci were closely apposed to cis -Golgi sites marked by PfGRASP–GFP, and upon treatment with brefeldin A they accumulated a model cargo protein via a process dependent on the presence of an intact diacidic export motif. Our data suggest that the cargo-binding function of PfSec24a is conserved and that accumulation of cargo in discrete tER sites depends upon positive sorting signals. Furthermore, the number and position of tER sites with respect to the cis -Golgi suggests a co-ordinated biogenesis of these domains.     

Self-maintenance of foci of bovine leptospirosis]

The relationship between the rate of cattle infection with leptospirosis and the total number of livestock at cattle-breeding farms has been established. The annual dynamics of this infection has been found to give two morbidity rises among the animals, occurring not due to their contacts with the natural foci of leptospirosis, but as a consequence of the animal vertical and horizontal "circulation". The mechanisms of self-maintenance of the foci of leptospirosis among cattle are discussed.     

Use of the indirect hemagglutination test for the study of ornithosis. Report 2. Preparation and approbation of the dry ornithosis erythrocyte diagnostic agent

An original preparation--dry ornithosis erythrocytic diagnostic agent for the indirect hemagglutination test was prepared on the basis of formalinized tannin-treated sheep red blood cells and group-specific phospholipid antigen of the causative agent of ornithosis. This diagnostic agent retained its specific activity for 18 months (observation period). The use of this preparation considerably facilitated the method of performance of this test, this offering a possibility of its wide application in practice. A sufficiently high sensitivity and specificity of the indirect hemagglutination test with the suggested diagnostic agent, in comparison with the complement fixation test was demonstrated.     

IMMUNOLOCALIZATION OF CENTRIN DURING FERTILIZATION AND THE FIRST CELL CYCLE IN FUCUS DISTICHUS AND PELVETIA COMPRESSA (FUCALES,PHAEOPHYCEAE)1

Antibodies that recognize the centrosome-associated protein centrin were used to characterize centrosomal origin and positioning during fertilization and the first cell cycle in Fucus distichus subsp. evanescens (C. Agardh) Powell and Pelvetia compressa (J. Agardh) De Toni. Centrin was identified in sperm, eggs, and zygotes on protein blots, indicating the protein is present in both gametes. Using immunofluorescence microscopy, centrin was found in discrete foci in sperm. In contrast, eggs lack centrosomes and centrin was not detectable by immunofluorescence, indicating that centrin was probably dispersed in the cytoplasm. Two foci of centrin were present on the nuclear envelope of zygotes, but microtubules remained dispersed over the zygotic nucleus. Centrin foci separated over the nuclear envelope as the first cell cycle progressed. Microtubules became concentrated at the centrin foci to form centrosomes that gave rise to the spindle poles at mitosis.     

Non-spindle microtubule organizing centers in metaphase II-arrested mouse oocytes

A human autoantiserum (5051) directed against pericentriolar material (PCM) was used to study the distribution of microtubule-organizing centers (MTOCs) in the oocyte and during the first cell cycle of mouse development. In oocytes, the PCM was found not only at the poles of the barrel-shaped metaphase II spindle but also at many discrete loci around the cytoplasm near the cell cortex. The spindle poles were also composed of several PCM foci. In metaphase-arrested eggs only the PCM foci located near the chromosomes acted as MTOCs. However, after reduction of the critical concentration for tubulin polymerization by taxol, the cytoplasmic PCM foci were also found to be associated with nucleation of microtubules. After fertilization the cortical PCM foci remained in a peripheral position until the end of the S phase, when they appeared to migrate centrally towards the pronuclei. At prometaphase of the first mitotic division, numerous MTOCs were found around the two sets of chromosomes; these MTOCs then aligned to form two bands on either side of the metaphase plate of the first mitosis.     

DNA damage response and cellular senescence in tissues of aging mice

The impact of cellular senescence onto aging of organisms is not fully clear, not at least because of the scarcity of reliable data on the mere frequency of senescent cells in aging tissues. Activation of a DNA damage response including formation of DNA damage foci containing activated H2A.X (γ-H2A.X) at either uncapped telomeres or persistent DNA strand breaks is the major trigger of cell senescence. Therefore, γ-H2A.X immunohistochemistry (IHC) was established by us as a reliable quantitative indicator of senescence in fibroblasts in vitro and in hepatocytes in vivo and the age dependency of DNA damage foci accumulation in ten organs of C57Bl6 mice was analysed over an age range from 12 to 42 months. There were significant increases with age in the frequency of foci-containing cells in lung, spleen, dermis, liver and gut epithelium. In liver, foci-positive cells were preferentially found in the centrilobular area, which is exposed to higher levels of oxidative stress. Foci formation in the intestine was restricted to the crypts. It was not associated with either apoptosis or hyperproliferation. That telomeres shortened with age in both crypt and villus enterocytes, but telomeres in the crypt epithelium were longer than those in villi at all ages were confirmed by us. Still, there was no more than random co-localization between γ-H2A.X foci and telomeres even in crypts from very old mice, indicating that senescence in the crypt enterocytes is telomere independent. The results suggest that stress-dependent cell senescence could play a causal role for aging of mice.