Regional differences of B cell immune responses to (4-hydroxy-3 nitrophenylacetyl) acetic acid (NP) antigen in senescent mucosal immune system. The age-associated primary immune response of B cells from the mesenteric lymph nodes (MLN), the bronchial lymph nodes (BLN), lamina propria (LP), Peyer's patches (PP), and the spleen (Sp) of mice following peritoneal or oral immunization with (4-hydroxy-3-nitrophenylacetyl) acetic acid (NP) conjugated bovine gamma-globulin (NP-BGG) was investigated. The induction of immune responses was assessed in 4-, 11-, and 24-month old individual C57BL/6J male mice by determining the number and isotype of anti-NP ELISPOT-forming cells (SFC) in the MLN, BLN, PP, LP, and Sp, and the titer and isotype of serum anti-NP antibody. Data indicated a significant age-associated decline in immunoglobulin M (IgM) and IgG anti-NP SFC in the Sp. No age-related declines were seen in the anti-NP SFC responses in the MLN and BLN. Oral immunization of mice with soluble NP-BGG in combination with cholera toxin resulted in anti-NP SFC responses in LP and PP, which were predominantly of the IgM class followed by IgG and IgA. There were age-associated increases in IgM, IgG, and IgA anti-NP SFC responses in the LP and PP with negligible responses in the SP and MLN. Serum anti-NP IgM and IgG antibody titers in mice immunized intraperitoneally with NP-BGG were found to decline with age. Isoelectric focusing and affinity immunoblotting revealed a loss of several anti-NP antibody clonotypes in the immune serum of old mice following parenteral immunization with NP-BGG as opposed to several new antibody clonotypes following oral immunization. The findings suggested age-related regional differences of B cell immune responses to the NP antigen in the mucosal immune system. These changes in the NP specific antibody repertoires in the spleen and mucosa-associated tissues may be regulated by different mechanisms operating towards different antigens, and this in turn may influence the characteristic antibody responding to these antigens.


Persistent pulmonary interstitial fibrosis, induced by immune response to TNP, is associated with altered mRNA procollagen type I:III ratio. Pulmonary interstitial fibrosis is characterized by a progressive increase in connective tissue in the lung parenchyma. Fibrosis is associated with conditions that result as a consequence of cell mediated responses including graft versus host disease, delayed type hypersensitivity reactions, and granulomas. The hapten-immune animal model for pulmonary interstitial fibrosis correlates the nonresolving fibrosis observed in the lung parenchyma directly with the animal's prior immunization to the hapten. Because the model is patterned after the well studied contact hypersensitivity assay in the skin, the immune response can be directly correlated with a cell-mediated (T-lymphocyte) immune mechanism. Previously, we reported that hapten-immune animals showed increased collagen deposition as identified on routine paraffin fixed slides that were stained with Masson's trichome. In this report, morphometric procedures were used to quantitate the fibrotic lesion. Fibroblasts were harvested from lungs of all treatment groups, cultured, and assayed for collagen production. Once it was determined that collagen production by fibroblasts was similar to that recorded in assays using fresh lung tissue, the fibroblasts were used as a homogeneous cell source for RNA. Total RNA from various treatment groups was used to assess the ratio of mRNA for procollagen I:III using slot and northern blot hybridization procedures. An increased ratio in the procollagen type I:III mRNA was observed in total RNA isolated from fibroblasts from immune and challenged hamsters, and not in samples from all other groups. These results support the hypothesis that the activated T lymphocytes involved in "contact" hypersensitivity-like reactions in the lung regulate not only the quantity, but also the quality of collagen produced by the fibroblasts in the lungs of the hamsters that develop nonresolving fibrosis. The model may be important for the study of skin and pulmonary disease induced by exposure to environmental haptens.


[Growth and development of Metorchis orientalis in chicks and its adult morphology]. In order to observe the infectivity, growth and development and adult morphology of Metorchis orientalis, a total of 40 chicks were experimentally infected with 100 metacercariae respectively, collected from Pseudorasbora parva. The worms of various developmental stages were recovered from chicks at 1.5, 3, 5, 7, 9, 11, 14 and 21 days after infection, and they were prepared for morphological observations and measurements. All of the worms were found in the gallbladders of chicks, and their recovery rate was 32% in average. The growth of the body was rapid from 9 to 11 days after infection. The genital primordia appeared in 1.5 and 3-day old worms, and ovary and testes were first observed in 5-day old worms. Thereafter, genital organs gradually matured and completed up to 11 days after infection. The adult worm was leaf-like, and possessed a convoluted tubular seminal vesicle, an ovoid ovary, a sac-like seminal receptacle, 2 lobed-testes and follicular vitellaria. Eggs were 31.9 x 15.3 microns in average size, ellipsoid to elliptical in shape and possessed abopercular thickenings. From the above results, it is concluded that M. orientalis grows in sigmoid pattern in chicks, and their genital organs fully mature between days 9 and 11. It is also confirmed that a chick is a new definitive host of M. orientalis.


Tonsillar (Waldyer's ring equivalent) lymphoid tissue in the rat: lymphocyte subset binding to high endothelial venules (HEV) and in situ distribution. We have studied lymphocyte traffic to the Waldeyer's ring equivalent (WRE) lymphoid tissue of the rat, by measuring the in vitro binding of various lymphocyte subsets to high endothelial venules (HEV) in the WRE. In addition, we studied the in situ distribution of these lymphocyte subsets. WRE tissue consists of B and T cell areas; the latter contain HEV. B cells outnumber T cells, and T helper (CD4) cells outnumber T suppressor/cytotoxic (CD8) cells (T/B ratio = 0.7; CD4/CD8 ratio = 5.1). In vitro studies of lymphocyte binding showed that lymphocytes adhere almost equally well to HEV in WRE tissue as to HEV in lymph node (LN) tissue, and much better to HEV in WRE than to HEV in Peyer's patch (PP) tissue. T cells bind better than B cells to HEV in WRE (T/B binding ratio = 1.8), and CD8 cells better than CD4 cells (CD8/CD4 ratio of 2.9-3.2, dependent on cell source). The observed preference of T over B cells in binding to HEV is not reflecting the distribution of these lymphocyte sets in situ. In this respect the WRE takes a unique position, since in other lymphoid organs T/B binding ratios parallels T and B cell distribution in situ. This may suggest a much more rapid passage of T cells through the WRE than through other lymphoid tissues, although other mechanisms cannot be ruled out. The CD8/CD4 binding ratio to HEV in WRE contrasts with situ distribution of these cells also; however, this is found for LN and PP lymphoid tissue too.(ABSTRACT TRUNCATED AT 250 WORDS)


Trematodes of the genus Haematoloechus (Digenea: Plagiorchiidae) from frogs in Korea. Total 242 Rana nigromaculata and 9 R. catesbeiana were collected from the various localities in Korea from February 1989 to July 1991, and their lungs were examined. Five species in genus Haematoloechus, i.e., H. sibiricus japonicus (Yamaguti, 1936), H. nanchangensis Hsiung, 1934, H. variegatus (Rudolphi, 1819), H. lobatus (Seno, 1907), H. lobatus koreanus, were identified in this study. Among them, H. lobatus koreanus was a new subspecies and H. variegatus and H. lobatus were newly recorded from Korea.


Suppression of resistance to Giardia muris and cytokine production in a murine model of acquired immune deficiency syndrome. Peyer's patch (PP) T cells through the production of appropriate cytokines foster the development of immunity to the intestinal protozoan parasites such as Giardia. T cell destruction by the human immunodeficiency virus precedes the development of acquired immune deficiency syndrome. Thus, HIV may increase susceptibility to intestinal parasite infections. Therefore, we measured the resistance and T cell cytokine responses to Giardia in C57B1/6 mice infected with the retrovirus LP-BM5 which produces a murine AIDS (MAIDS). Mice with MAIDS and controls were intragastrically challenged with 1 x 10(5) G. muris cysts. Fecal counts were measured weekly following challenge. Also, PP T cell production of interleukin (IL)2, IL3, IL4, and Interferon-gamma in response to G. muris trophozoite antigens displayed on antigen presenting cells were measured at these times. Prior to day 14 of the infection, the number of Giardia cysts in the retrovirus group paralleled that in controls. However, by day 21 after Giardia infection, mice with MAIDS failed to clear the Giardia cysts from the intestine while the control mice were completely free of cysts. IL2 and IL4 production in response to Giardia trophozoites by unfractionated PP lymphocytes were severely depressed in the retrovirus infected group, while IFN-gamma production was increased. Depressed cytokine production was most likely due to depressed PP T cell numbers. When fractionated enriched T cells were adjusted to a uniform concentration in in vitro immunization cultures, the production of IL2 and IL4/IL5 were similar between retrovirus infected compared with control mice. Recoverable PP T cells were lower in mice with MAIDS.(ABSTRACT TRUNCATED AT 250 WORDS)


[Aporocotyle theragrae (Trematoda: Aporocotylidae) from the blood vessel of Theragra chalcogramma]. Two fully matured specimens were collected from the blood vessel of two fish, Theragra chalcogramma, which was bought at the Emun market of Seoul in May, 1985. The blood fluke has no pharynx and suckers. Its body surface was covered with minute spines forming fan-shaped groups, 15 microns long. The intestine was H-shaped, and the ratio of esophageal length to body length was 1:6.1-6.8. Irregular shaped testes were 109-114 in number, occupying the intercecal space from cecal bifurcation to the genital pore. The present species was identified as Aporocotyle theragrae. This is the first report of the species in Korea.


[Experimental Cryptosporidium parvum infection in a Korean native calf isolated from a Korean mouse]. This study was performed to investigate experimental transmission of Cryptosporidium parvum in a calf. A 25-day-old Korean native calf was inoculated per os with 1 x 10(6) C. parvum oocysts isolated from a Korean mouse. The calf commenced oocyst discharge in feces on post-inoculation day 4, and continued until the day 11. The number of discharged oocysts peaked (4.9 x 10(5)) on post-inoculation day 6. However, the calf did not show signs of diarrhea. The present results indicate that C. parvum is cross-transmissible between the calf and the mouse.


[Infection status of larval anisakids in Astroconger myriaster collected from the Southern Sea near Pusan]. A study was presented on the anisakid larvae in Astroconger myriaster which were caught at the Southern Sea and saled at Pusan area. The nematodes were morphologically classified into Anisakis sp. and Contracaecum sp. after fixation in 70% ethanol and clearing in lactophenol. Total of 1,768 larval anisakids were collected from 259 (67.8% positive rate) out of 382 examined A. myriaster. Total 642 larvae of Anisakis sp. were obtained from 94 (24.6%) and 1,126 Contracaecum sp. were recovered from 165 (43.1%) fish. The average number of worms per infected fish was 4.6. The infection rate increased according to the length of fish, and all of the fish over 71 cm were found infected. The numbers (proportions) of recovered worms by the organs were 1,440 (80.5%) in the omentum, 166 (9.4%) in the intestine, 107 (6.0%) in the stomach, 32 (1.8%) in the skin, 18 (1.0%) in the muscle, 13 (0.7%) in the liver, and 2 (0.1%) in the head. The larvae in the muscle may infect the humans who are enjoying raw sliced meat of the fish.


Experimental infection of pigs and cattle with eggs of Asian Taenia saginata with special reference to its extrahepatic viscerotropism. Asian Taenia saginata, tentatively called Taenia saginata taiwanensis, has been described to be infected in its metacestode stage only in the liver of intermediate host animals. Experimentally, however, we found that the metacestodes of the Asian Taenia saginata are also infected in other viscera than the liver of pigs (Landrace-Duroc-Hampshire) 4 days to 4 months postinoculation (PI). Viscerotropism of cysticercosis was apparent because a majority (70.7%) of the non-calcified cysticerci were found in the livers while a minority were found in extrahepatic organs such as the omentum (19.2%), lungs (8.1%) and serosa of colon (2.0%). When experimentally infected to cattle, Asian T. saginata cysticerci were also observed calcified in the livers. On the other hand, classical Taenia saginata metacestodes infected the muscles and viscera of the Holstein-Friesian cattle whereas no infection was observed in experimental pigs. Extrahepatic metacestodes of Asian T. saginata, which were obtained from an experimental pig were confirmed to be infective to a male volunteer. This extrahepatic viscerotropism of Asian T. saginata metacestodes in experimental pigs explains well the transmission modes of Asian T. saginata among people considering the eating habits.


Susceptibility of various animals to Pneumocystis carinii infection. Pneumocystis carinii (Pc) is an important opportunistic pathogen of immune compromised hosts, and is known to infect various animals. The present study observed the infection status of 6 mammals and 3 strains of albino rats with Pc after suppression of their immunity. Methyl-prednisolone was injected once a week and tetracycline was supplied with water for 5 to 21 weeks. Hamsters, guinea pigs, rabbits, dogs, cats and pigs were negative by impression smear, and only the rats were found infected by Pc. All of the three strains of rats, Sprague-Dawley(SD), Wistar(W) and Fisher(F), were infected by Pc but W rats showed heavier degree of infection in earlier period than F or SD rats. The present findings suggest that W rat is the best among the animals used in the present study for production of Pc.


Transmission modes of Pneumocystis carinii among rats observed by karyotype analysis. To observe the transmission patterns of karyotype of Pneumocystis carinii (Pc) by rat colonies, three strains of rats, Sprague-Dawley(SD), Wistar(W) and Fisher(F) from various animal vendors, were suppressed of their immunity by injection of methyl prednisolone. They were kept for 5 to 13 weeks in 3 different animal rooms, A, B, and C. The purified organisms were prepared in low melting point agarose gel by embedded lysis method for pulsed field gel electrophoresis. Field inversion gel electrophoresis showed 2 patterns of the karyotype of Pc. The rooms A and C contained SD rats from the source P, and also the room A was used for F and W rats. However, Pc from all of the SD and F rats in the room A showed same karyotypes, the pattern I. The SD rats from different vendors, M and S, were reared in the room B, and shared the same Pc karyotypes, the pattern II. The rats of W strain were from the vendor M, and immune-suppressed in the animal room A. Five weeks after the experiment, the Pc showed the karyotype pattern II but the pattern became mixed with the type I after 7 to 8 weeks. The findings revealed that the animals born and reared in the same animal quarter harbored Pc with same karyotypes. If the animals were kept under immune-suppression in the same room with heavily infected hosts, they could be infected by Pc from their neighbors. The present experimental findings suggest that Pc is transmitted among rats through the air.


[Immunological approach for classification of free-living amoeba in Korea]. Acanthamoeba spp., free-living amoebae inhabited in moist soil, pond, freshwater, sewage, atmosphere and swimming pool, may be causative protozoa of the fatal primary amoebic meningoencephalitis in experimental animals and humans. In this study, Acanthamoeba spp., including Acanthamoeba sp. YM-4 (isolated strain from Korea) had been compared by the two-dimensional electrophoresis and hybridoma technique as well as the difference of morphological characteristics. Trophozoite of Acanthamoeba sp. YM-4 is usually uninucleate and show the hyaline filamentous projections (acanthopoda). No flagellate stage observed. Cysts have two walls, the outer wall is nearly circular, but inner wall is oval or some irregular. As results of SDS-PAGE for lysate of Acanthamoeba sp. YM-4, 16 major protein fractions are similar to those of A. culbertsoni, but different to A. royreba and A. polyphaga. Findings of two-dimensional electrophoretic patterns of Acanthamoeba sp. YM-4 are almost same to those of A. culbertsoni, The isotype of monoclonal antibodies produced from McAY 6, McAY 7, McAY 8, McAY 13 and McAY 16 clones were IgG1, and McAY 10 and McAY 11 clones were IgM. As results of the cross-reactivity among various amoebae using ELISA with monoclonal antibodies, McAY 7 monoclonal antibody (molecular weight 43 kDa by EITB) was only reacted with Acanthamoeba sp. YM-4, but McAY 6 and McAY 10 monoclonal antibodies were reacted to A. culbertsoni as well as Acanthamoeba sp. YM-4.


Measurement of 150 kDa protein of Taenia solium metacestodes by antibody-sandwich ELISA in cerebrospinal fluid of neurocysticercosis patients. An antigenic protein in cystic fluid of Taenia solium metacestodes (CF) of 150 kDa was measured by antibody-sandwich ELISA in serum and cerebrospinal fluid (CSF) of neurocysticercosis patients. Capture antibodies were rabbit antisera against CF (RACF) and a monoclonal antibody (MAb) against 150 kDa protein in CF. Lower limit of antibody-sandwich ELISA was 8 ng/ml of the protein. Except CF, no tested helminths extracts reacted. Levels of the protein in 351 sera from 255 patients (55 surgery confirmed and 202 antibody and CT/MRI confirmed) were below sensitivity of the assay. Of 276 CSF from 212 patients, 31 samples (11.2%) showed positive findings. This assay, therefore, was not sensitive enough to be a diagnostic. Instead, the 150 kDa protein appeared in CSF in such situations as in 2 days after praziquantel treatment, or as in a patient infected with a racemose cysticercus with degenerated cyst wall. Of cases whose follow-up CSF were assayed, 2 cases showed that the protein appeared intermittently. These results suggest strongly that appearance of free 150 kDa protein is associated with cyst wall rupture. In CSF which contained the 150 kDa protein over 61 ng/ml, the protein was recognized in SDS-PAGE before and after immunoprecipitation.


[Antigenic localities in the tissues of Metagonimus yokogawai in the period of growth]. In order to observe the antigenic localization in the tissues of Metagonimus yokogawai in growth stages, immunogoldlabeling method was applied to using serum of the cat which infected with isolated metacercariae from Plecoglossus altivelis. The sectioned worm tissues from each growth stages were embedded in Lowicryl HM 20 medium, stained with infected serum IgG and protein A gold complex (particle size: 12 nm) and observed by electron microscopy. In the worm tissues of all experimental groups, the gold particles were specifically concentrated on the tegumental syncytium and cytoplasm of the tegumental cell as well as the secretory granules in the parenchymal tissue. In the 16th and 20th week grown worm tissues, the gold particles were specifically concentrated on the vesicles in the tegumental syncytium and cytoplasm of the tegumental cell. The gold particles were specifically concentrated on the caecal epithelia of the 4th, 8th and 12th week growth groups but slightly concentrated on those of the 16th and 20th week.


[Comparative antigen analysis of Trichomonas vaginalis by enzyme-linked immunoelectrotransfer blot technique]. Analysis of six isolates of Trichomonas vaginalis was carried out with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and enzyme-linked immunoelectrotransfer blot (EITB). Trichloroacetic acid-treated antigens of the 6 isolates revealed 25 protein profiles ranging 12-170 kDa of molecular weight in SDS-PAGE. In EITB, the specific immunogenic bands were visualized at 51 kDa and 96 kDa when HY-1 antigen was probed with different mice sera immunized with 6 isolates of T. vaginalis. The banding patterns with different sera showed isolate-to-isolate variability. In EITB, homologous antigen (HY-1) did not show any enhanced response in reacting to homologous antiserum (HY-1) when 6 isolates of T. vaginalis were probed with a single serum (HY-1). It is assumed that the different banding patterns of six isolates show isolate-to-isolate variability and immunogenic common bands in 41, 47, 74 and 94 kDa on EITB may connote the important significance on immune response in T. vaginalis infection.


Study on vector mites of tsutsugamushi disease in Cheju Island, Korea. Because no reference on trombiculid mites (Acarina: Trombiculidae) in Cheju Island where tsutsugamushi disease is highly endemic had been available, studies on trombiculid mites in Cheju Island were implemented during the period of August 1991-April 1992, and the results obtained are summarized as follows: (1) The species and numbers of the field rodents collected were 143 Apodemus agrarius chejuensis (92.3%), 11 Crocidura lasiura (7.1%) and 1 Micromys minutus (0.6%). From total 12,075 chiggers harvested, 9 species of 4 genera in Trombiculidae were identified. (2) The predominant species through all seasons was L. zetum (43.3%), followed by L. orientale (27.4%) and L. scutellare (26.6%). However, in autumn when the most cases of tsutsugamushi disease occur, L. scutellare was prominently predominant, having 79.8% of the collected chiggers. (3) Among 1,142 L. scutellare examined for Rickettsia tsutsugamushi by means of IFA test, 6 individuals were found positive showing 0.5% of infection rate. This is the first finding that L. scutellare is the second vector species of tsutsugamushi disease in Korea. (4) Antibody positive rate of A. agrarius chejuensis sera were 31.2% (44/139), and 1 M. minutus serum was also found positive. The seropositive rates by season were not so significantly different.


Mass treatment of head louse infestation with Sumithrin powder in primary schools in Korea. A mass treatment of head louse infestation with Sumithrin powder (0.4% phenothrin) in primary school children was implemented during the period of September 1991-May 1992. The infestation rate of total 2,515 children was 38.6% in average (21.2% in boys and 57.2% in girls). The reduction rate of head louse infestation was 93.4% with a single treatment and 94.8% with double consecutive treatments with about 10 days interval, which indicated that a single treatment would be recommended for the mass treatment in the community. Long term follow-up after Sumithrin powder application for head louse control in a primary school showed that the infestation rate dropped from 33.1% before treatment to 5.4% by seven months after treatment, giving a 83.4% reduction rate.


Telemicroscopy. Design considerations for a key tool in telepathology. The paper introduces telemicroscopy as a mean to establish online telepathology services. It is differentiated between specialized point to point connections with only a few restrictions on one side, and telecommunication links available inexpensive worldwide in the form of the public telephone net on the other side with the restriction of a limited channel capacity. The restrictions imposed by the application of the telephone net (preferentially the ISDN service) are discussed and strategies outlined to overcome these restrictions. It is concluded, that if dedicated broadband connections are available, realtime telemicroscopy links can be realized immediately with available equipment. If a switched and general available telemicroscopy service applying public telecommunication links is favoured, for effective and acceptable ("online") links, computer intelligence has to be introduced to allow effective strategies for data reduction and data transmission. In this field design and development efforts are still necessary.


Aspects of standardization in telepathology. Effective telepathology is only possible, if the partners are supplied by a broad variety of equipment connected by far-spread communication lines. These prerequisites can only be fulfilled if technical and medical standards are available obligate for all participants. The technical and medical aspects of those standards are described in general and include procedures and notations in pathology, standards of the image source, image sampling, clinical information, classification of diagnosis, communication channels, hard- and software equipment. New standards should only be developed for those not existing, or if the already existing ones are not appropriate. The standardization classes should allow flexible formats and be developed as "open modules" allowing "individual" peripheral work stations and progressive development in the future.


Legal aspects of telepathology.

Legal aspects of telepathology. The legal aspects of telecommunication are discussed briefly. Telepathology induces no new legal conditions; however, due to its somewhat higher diagnostic error rate specific problems may occur. Both the pathologist and hospital should clarify the internal liability conditions. The pathologist should be aware that he is still liable for errors induced by the technician, even without having the possibility of responsibility or any supervision.


Visual telecommunication for expert consultation of intraoperative sections. Histopathological images of intraoperative sections were transferred by use of normal telephone line between the Department of Pathology, Thoraxklinik, Heidelberg, and the Institute of Pathology, Hospital Baumgartnerhöhe, Vienna. The images were digitized using a colour TV camera and stored in a specific image transmission system (VP2000). The transfer of one image lasted 1.4-2 min at average; normally two different areas of the section were transmitted. The expert discussion including the clinical history usually started a few minutes prior to the image transfer, and was finished 3 min after the transfer of the last image. A substantial assistance in the intraoperative classification of the disease was achieved in 37% of the 35 submitted cases. The images transfer was only useful for medium to high magnifications of the microscope, i.e., an objective x 25 or higher. The trail allows the following conclusions: a) Visual transfer of microscopic images is a useful technique for expert consultation of intraoperative sections, and can give additional information in 20-40% of the analyzed cases; b) it is useful only for medium or high microscopic magnifications and limited by the spatial resolution of the TV cameras.


Human performance studies of the video microscopy component of a dynamic telepathology system. Human performance studies were performed to evaluate the video microscopy component of a proposed dynamic telepathology system. Frozen sections from breast biopsies of 115 patients were evaluated by both conventional light microscopy and video microscopy by 6 pathologists. Receiver operator characteristic (ROC) curve studies showed nearly identical levels of discrimination between benign and malignant breast lesions for both viewing modalities. Viewing times were significantly greater (p < 0.001) for video microscopy but within a time frame that rendered the technology of potential value for pathology diagnostic applications. Large degrees of interobserver viewing time variability were found for light and video microscopy.


The validity of frozen section diagnosis based on video-microscopy. To investigate the accuracy of video-microscopy of frozen sections, two pathologists reexamined 80 cases of archival material, on which frozen sections had previously been performed. Diagnoses based on the two trials (Observer 1 and 2) and diagnoses obtained in the original frozen section situation were compared with the final diagnoses based on paraffin embedded material. Observer 1 had two false negative diagnoses, but no false positive, whereas Observer 2 had one false positive, but no false negative diagnosis when compared with the final diagnoses. No false positive or false negative diagnoses were made in the original frozen sections situation and the number of inconclusive diagnoses were 5, as compared with 6 and 8 in the two trials based on video-microscopy. More experience with video-microscopy will probably achieve a quality of frozen section diagnoses similar to that of direct light-microscopy.


Frozen section service via the telenetwork in northern Norway. We present preliminary results of remote frozen section service for two local hospitals in Northern Norway. The service is arranged by remote controlling microscopes with motorized X, Y and Z stage movements, magnification and illumination located at Kirkenes and Harstad Hospitals at a distance up to 400 km apart from the workstation at the University Hospital in Tromsø. The video-images of the frozen section are transmitted via a two-ways phone and video telenetwork with 2 Mbit/s capacity. The images are displayed on a monitor as both still and live images and diagnosed by pathologists in Tromsø. To data, 50 patients are examined by remote frozen section service. Correct benign versus malignant diagnoses are given in all cases compared with final diagnoses based on formalin fixed and paraffin embedded material except for two false negative malignant cases and two deferred diagnoses. The average time taken for examining each case of frozen section was 13 minutes. For hospitals with limited requirement of local pathology service and for hospitals with deficiency of specialists, remote frozen section service may be a worthwhile substitute.


Experience and present status of telepathology in the National Cancer Center Hospital, Tokyo. The optical fiber high definition television (HDTV) live image system with a remote control microscope is an ideal telepathology system. Diagnostic accuracy with this system is high enough for pathology consultation. However, it is very expensive to set up. The integrated services digital network (ISDN)-HDTV still image system using public telephone lines presently available in Japan INS 64 (64 kb/s), is not suitable for frozen section diagnosis and diagnosing difficult lesions. However, it functions very well for cytology consultation. When INS 1500 (1.5 Mb/s) becomes available in practice, it can substitute the optical fiber-HDTV live image system in terms of cost and efficacy. Pathologists must become accustomed to making diagnosis from images on the monitor. They must be very careful in making diagnosis, when images are transmitted from institutions without pathologists, since selection error may occur at the time of frozen section and at the time of transmission of images.


Telepathology: a new tool of pathology? Presentation of a French national network. The field of pathology is constantly growing and the responsibility of the pathologists has progressively increased in the decision of application of some efficient but aggressive therapies. They justify the increasing necessity of discussion between pathologists about the diagnoses of difficult or rare or severe lesions. It is now possible to discuss at distance on good definition digitized images transmitted through I.S.D.N. A French experimental network between 12 workstations has been introduced since 2 years. Difficult or severe diagnoses have been immediately solved in 85% of the discussed cases. It remain legal problems and the necessity of an evaluation protocol which could be discussed at a European level. The increase of technical progress and, above all, the decrease of the costs might favour the development of telepathology. The workstation of the pathologist in the year 2000 could be the combination of a microscope, a HD-TV-camera and a personal computer with many softwares allowing treatment and transmission of numerous pictures.


Telepathology in Greece. Experience of the Metaxas Cancer Institute. This paper deals with the description of a telecommunication project which is initiated by Metaxas Pathology Laboratory. In the first section we discuss a comprehensive computer supported information system which was installed in the laboratory. Special emphasis is given at the design of the communication subsystem. Then, the configuration of the diagnostic equipment is described. The last section of the paper focuses on the discussion of the experimental use of an integrated "low cost" system for interconnection among the Metaxas Pathology laboratory, the Pathology Department of the University of Crete, and the Technological Educational Institute (TEI) of Athens.


Telepathology in Sweden. A national study including all histopathology and cytology laboratories. Quality improvement and standardization of diagnosis in histopathology and cytology are important for the future of the discipline. Nominal scale diagnoses dominate the practice and their standardization depends on relevant and reproducibly identifiable criteria as well as on communication of these among pathologists. Telepathology, i.e. the transmission of adequately detailed colour images of microscopic fields over the telephone network is now a realistic possibility. All 30 laboratories for histopathology and cytology in Sweden will have access to Telepathology work stations for at least 8 weeks each during 1992-1993. Two centers will have permanent stations from September 1992. The images will be transmitted over the ISDN network, allowing a compressed image to appear instantaneously. This image is then gradually and imperceptibly decompressed during 15-60 seconds, the time depending on the complexity of the image. A program for consultation and quality testing is being set up, which will be evaluated during 1993. Based on a recognition of the conditions for diagnosis in pathology and cytology indicated above, the Swedish Society of Pathology has initiated a project called "Telepathology in Sweden". It is a joint effort with active participation by Swedish Telecom and the Swedish Planning and Rationalization Institute for the Health and Social Services, Stockholm as well as by Innovativ Vision AB, Linköping, a company providing hard- and software for image analysis, image banks and communication.


Visual documentation of neoplasms as part of the REGISTRY Nomenclature Information System. The REGISTRY Nomenclature Information system (RENI) is a program which runs on IBM-compatible personal computers. It includes internationally accepted diagnostic criteria for tumors and pre-neoplastic lesions in the rat to be used in toxicologic pathology. A video disk system, connected to the computer, provides colored pictures of histopathological reference slides, which are used for a visual documentation of the written criteria.


Cloning of par region and the effect of par region on the stability of pUC9. The par region of pSC101 was cloned into pUC9, and pEC302 was obtained. The stability of pEC302 and pUC9 in E. coli HB101 was tested on M9 medium to determine the effect of par region on the stability of plasmid. After 40 generations, pUC9 was almost completely lost, while pEC302 was 100% maintained in host cells.


The establishment of rat-mouse hybridomas secreting anti-aflatoxin M1 antibodies and the properties of their monoclonal antibodies. After a comparison of anti-AFM1-BSA antibody responses between rat and mouse, the spleen cells of rat with stronger responses were chosen as parent cells for fusing with mouse myeloma cells P3X63-Ag8.653. Through HAT medium selection, RIA screening and cloning, five well growing rat-mouse hybridoma clones were obtained that could secret anti-AFM1 antibodies stably. The results from ELISA and competitive binding RIA further proved that the 5 McAbs are direct against AFM1, with significant cross reaction to its derivative, AFB1. The average affinity constant of the 5 McAbs is 10(9)-10(11) l/M. It signifies that these monoclonals have potential application value for the construction of AF detection kit.


Mass culture of anchorage-dependent animal cells with newly born calf skin collagen membrane and microcarrier. It was shown that collagen substrate enhances the growth as well as the differentiation of many cells in culture. Collagen is one of the major fibrous proteins of animal bodies and the pure collagen used in this experiment was extracted from the newly born calf skin by chemical and biochemical methods. The analytical results obtained by ion-exchange chromatography and electrophoresis showed that the main components of denatured collagen were alpha monomers and beta dimers. The collagen and denatured collagen membranes were prepared by coating their solution on peteri dishes. Various types of cells were cultured on these membranes after being irradiated by ultraviolet ray. The denatured collagen was proved a good substratum for culturing anchorage dependent cells. A denatured collagen (gelatin) microcarrier, GT-2 was obtained by cross-linking gelatin with glutaraldehyde in a suspension polymerization process. These microcarriers were used successfully to culture various anchorage-dependent cells such as Vero, CHO, Bowes and fish cells in varying scales, including T-flasks, spinning bottles, revolving bottles and 1.5 l and 20 l bioreactors.


A basic study on the hybridoma cell culture of microencapsulation. We report here that the constitute component of Na alginate is a very important factor on microcapsular strength. We developed a method of estimating viable cell count in batch microcapsular process and the factors influencing the full loading rate in microcapsules. The physical integrity of microcapsule and full loading rate were greater than 95%. The cell line of C3 hybridoma was found to reach maximum intracapsule cell density of 8.8 x 10(6) - -2.1 x 10(7) cells/ml during static culture period of 6-10 days. During this period, the mouse monoclonal antibody accumulated in the intracapsular space to a final concentration of 379 micrograms/ml. Analysis of intracapsular protein by SDS gel electrophoresis during the culture period demonstrated that there was one band or two bands of protein which means its purity is relatively high.


Location and trans-action of the pac regulatory gene. The study on the localization of the pac gene indicated that the regulatory and structural genes were located in a 3.5 kb HindIII-EcoRI fragment. In this paper we report that a series of deletion derivatives of plasmid pPA6 was constructed and the effect of deletions on the pac expression was determined. The results suggested that the pac regulatory gene was located in the SphI-PvuII fragment within the pac structural gene. Then the SphI-PvuII fragment was cloned to the plasmid pTZ18U at the SphI-SmaI sites and the resultant plasmid pPA57 was transformed into the penicillin acylase producing strain E. coli D816 to observe the effect of SphI-PvuII fragment on the chromosomal pac expression. The results demonstrated that the regulatory gene within the SphI-PvuII fragment regulated the pac expression in trans. Computer analysis indicated that there were two ORFs in the SphI-PvuII fragment as possible candidates coding for the regulatory protein. The study on precise localization of the pac regulatory gene is in progress.


High-level expression of recombinant protein A in Escherichia coli. A high-level expression plasmid pPA-3 was constructed, which yielded up to 20% of soluble cell proteins as recombinant Protein A in E. coli DH5 alpha strain by heat shock induction. The recombinant Protein A contained only the five domains of native Protein A that bind with the Fc part of IgG. The molecular weights of the recombinant Protein A expressed in E. coli were determined to be 33kDa, 32.2kDa, 29.5kDa and 28.6kDa by SDS-PAGE and Western-blotting. The diversity of the molecular weights may be due to proteolysis, and a possible cleavage site is proposed. Some of the protein was purified with coupled human IgG by one-step affinity chromatography. The reactivity of the protein was compared with that of native Protein A and found that this protein could bind with more IgG than native Protein A in equal quantities of proteins.


Construction of a genetically engineered strain producing propionylspiramycin. Our work has demonstrated the cloning of propionyl acylase gene and the expression of S. mycarofaciens mutant in S. lividans TK54. In this paper, we report the transformation of pIJM9 recombinant plasmid containing the propionyl acylase gene into spiramycin producer S. spiramyceticus. The results of colony hybridization and Southern hybridization showed that No. 61 transformant harbored the pIJM9 recombinant plasmid. TLC and bioautography showed that the Rf value of one component of the fermentation products of No. 61 transformant was similar to that of propionylspiramycin. The HPLC retention time of the components of the fermentation products of No. 61 transformant and that of propionylspiramycin were also similar. Mass spectrum analysis showed that there was propionylspiramycin II in the fermentation products of No. 61 transformant. According to these results, No. 61 transformant is shown to be a genetic engineered strain producing propionylspiramycin.


G-banding analysis of radiation-induced chromosome damage in lymphocytes of Hiroshima A-bomb survivors. The present report describes the G-band analysis of somatic chromosomes in lymphocytes from 63 A-bomb survivors in Hiroshima to determine the type and frequency of radiation-induced chromosome aberrations. (1) The cells with stable-type chromosome aberrations (Cs cells) predominated among the aberrant cells, and showed a dose-dependent increase. All stable chromosome aberrations were classified into nine categories: reciprocal translocations, translocations of complex type, insertions, complex exchanges, peri- and paracentric inversions, terminal and interstitial deletions, and unidentified rearrangements. The frequencies of aberrations were found to increase with increasing dose for all aberration categories. Reciprocal translocations predominate in all dose ranges and among the chromosome aberrations classified. (2) The linear model was fitted to test the dose-response relationship for Cs cell frequencies. Employing a constant neutron RBEs of 10, an estimated linear slope of 15.2%/Sv was obtained for DS86 bone marrow dose with an intercept of 2.9% at dose 0. (3) Statistical analysis of data on 3,370 break sites showed good correlations between relative DNA content and the distribution of chromosome breaks involved in translocations, although the involvement of chromosome 1 is significantly higher.


Expression of the Tn antigen on erythroid cells from a patient with Tn syndrome. In order to examine expression of the Tn antigen on erythroid cells from a patient with Tn syndrome, we applied a selective two phase liquid culture system for human erythroid progenitors in peripheral blood. The cells were analyzed with flow cytometry employing an anti-Tn antibody and a lectin of Vicia villosa which recognizes only the Tn determinant. In the second phase, the Tn antigen was expressed on the cultured cells from the patient on day 3 and Tn-positive cells reached 62.7% on day 9. On the other hand, Tn-positive cells were not detected in the volunteer's cultured cells. When the patient's cells were co-cultured with the cells from a healthy volunteer, the percentage of Tn-positive cells was much lower than the expected value, suggesting that the normal cells suppressed the expression of Tn antigen on the patient's cells.


Allotypes of the fourth component of complement in Koreans. The analysis of genetic polymorphism in C4 was performed on EDTA-plasma from 169 healthy unrelated Koreans. Plasma samples were subjected to high-voltage agarose gel electrophoresis followed by immunofixation. C4B allotypes were further detected by a hemolytic overlay method. The allele frequencies of C4A and C4B were as follows; for C4A, C4A*3 = 0.6099, C4A*4 = 0.1702, C4A*Q0 = 0.1525, C4A*2 = 0.0461, and C4A*R = 0.0213; for C4B, C4B*1 = 0.6406, C4B*2 = 0.2740, C4B*5 = 0.0569, C4B*Q0 = 0.0178, and C4B*R = 0.0107. C4A*3 and C4B*1 were among the most common alleles at each locus. C4A*6 was not detected in this study, but this allele is relatively common in both Caucasoid and Negroid populations. C4B*5 is a common allele in Asian, which is rare in Caucasoids and Negroids. C4B*5 appeared to be a characteristic allele of Oriental. In the C4A locus, five individuals with duplicated allotypes (three C4A 3,3 + 2, one C4A 4,3 + 2, and one C4A 3,3 + 3) were observed, and in the C4B locus, one individual with duplicated allotype (C4B 1,1 + 1) was detected.


Population studies of INV(9) chromosomes in 4,300 Japanese: incidence, sex difference and clinical significance. Population incidence of a chromosome 9 with an inversion of qh, inv(9), was surveyed using C-banding in a total of 4,367 Japanese which consisted of five patient groups and a normal control group. The inv(9) incidence was 1.65% in the normal control group (n = 1,513) and 1.52% in the Down syndrome patient group (n = 1,246). The incidence of female carriers was 1.7 times higher than that of male carriers in the above two groups. The sex difference was significant (p < 0.05). Moreover, the incidence in the 47,XXY male group (n = 277) was slightly higher than female carrier incidences. The excess of carriers about 2 times was observed in the habitual abortion couple group (n = 694) and the spontaneous abortus group (n = 181), both having normal karyotypes. The present data probably indicate the existence of a genetic effect of inv(9) on human reproductive performance, while they could not explain the sex difference found in inv(9) carriers. Our speculation for the sex difference has been presented.


Localization of PMP-22 gene (candidate gene for the Charcot-Marie-Tooth disease 1A) to band 17p11.2 by direct R-banding fluorescence in situ hybridization. We mapped PMP-22 gene, candidate gene for the Charcot-Marie-Tooth disease (CMT) 1A, by direct R-banding fluorescence in situ hybridization. The signals of PMP-22 probe were localized to chromosome band 17p11.2. The present result was within the map position of the CMT 1A gene by genetic linkage analysis, and strongly indicated that PMP-22 gene is a candidate gene for the CMT 1A.


Localization of 24 cosmid clones on the human Y chromosome. Twenty-four novel cosmid clones were cloned and mapped on the human Y chromosome using a panel consisting of DNA from seven individuals each having a different segment of the Y chromosome. Eight were assigned to the short arm, 15 to the long arm and 1 to the both short and long arms.


Sex-determining region Y (SRY) in a patient with 46,XX true hermaphroditism. Using a polymerase chain reaction method, a search for Y-specific DNA sequences was made in samples derived from tissues of a 46,XX true hermaphrodite. We found a sequence of SRY in the ovotestis, skin and leukocytes. Other DNA sequences, which covered the pseudoautosomal boundary region, amelogenin gene and DYZ1 locus of Y-chromosome were not detected. The SRY gene detected in the patient by the polymerase chain reaction was not detected by Southern blot analysis, using the SRY fragment as a probe. These findings suggest that in the patient there is a mosaicism of cells with and without part of the Y chromosome, including the SRY sequence. As the SRY sequence was responsible for the development of the gonadal primordium to the ovotestis, SRY seems essential for gonadal differentiation in testis development.


Localization of pro-alpha 2(V) collagen transcripts in the tissues of the developing mouse embryo. Correct assembly of fibrillar collagen networks plays a critical role in animal morphogenesis. Very little is known about the contribution of the so-called minor fibrillar collagens (types V and XI) to fibrillogenesis. Here we examined the developmental expression of the mouse pro-alpha 2(V) collagen gene (col5a2) after the cloning and sequencing of cDNAs that cover the entire length of the message. Transcripts of col5a2, detectable as early as 9 days of gestation, localize with distinct patterns in the tissues of day 12.5 and day 16.5 fetuses. The earlier developmental stage is characterized by low and diffuse col5a2 expression in the peritoneal membranes and intestinal and craniofacial mesenchymes. The later stage exhibits higher and more restricted col5a2 mRNA accumulation in primary ossified regions, perichondrium, joints, tendon, atrioventricular valve of the heart, and selected portions of the head. A parallel analysis using a cartilage-specific pro-alpha 1(II) collagen (col2a1) probe confirmed that these two collagen genes are transcribed in a mutually exclusive manner during mouse embryogenesis. On the other hand, the developmental pattern of col5a2 expression closely resembles that of the type I collagen, thus further substantiating the notion that these macromolecules cooperate in the formation of fibrillar networks in non-cartilaginous matrices.


Localization of epidermal growth factor and its receptor in mandibular molars of the rat prior to and during prefunctional tooth eruption. Immunoperoxidase localization of epidermal growth factor receptors (EGFR) and epidermal growth factor (EGF) itself was examined in rat first and second mandibular molars postnatally from day 0 to 12. The results showed that the dental follicle stained heavily for EGFR from day 0 to 8, declined in staining at day 9, and was devoid of stain from day 10 onward. Preosteoblasts and osteoblasts of alveolar bone also stained and lesser staining of ameloblasts and odontoblasts was observed. Except for staining of occasional isolated cells, the stellate reticulum did not stain. Light staining of the dental pulp of the first mandibular molar was seen from day 0 onward but the pulp of the second molar did not stain until approximately day 6. With respect to EGF, the dental follicle also stained for it until day 12. The ameloblasts stained more intensely for EGF than for EGFR. Because injections of EGF cause premature eruption of teeth and because the presence of a dental follicle is necessary for eruption, this study suggests that EGF could have its effect on the follicle as seen by the presence of EGFR receptors on the follicle. Moreover, because EGF exerts its effects early (day 0-3) to cause eruption and because the influx of monocytes into the follicle to form osteoclasts for bone resorption for eruption occurs early, the heavy staining for EGFR in the follicle early followed by the absence of staining at day 10 correlates chronologically with the key molecular and cellular events of eruption. Finally, the presence of EGF in the follicle, as well as enamel organ, could provide an endogenous source of EGF to regulate tooth eruption, either by an autocrine or a paracrine effect. Thus, the localization of EGFR and EGF in the dental follicle coupled with the chronology of localization suggests that EGF could play a physiological role in tooth eruption.


Noncoordinate developmental regulation of N-cadherin, N-CAM, integrin, and fibronectin mRNA levels during myoblast terminal differentiation. N-cadherin, N-CAM, fibronectin, and beta 1-integrins have been implicated in the control of myoblast fusion to form multinucleate myotubes, a critical step in the terminal differentiation of skeletal muscle. We have analyzed the temporal pattern of expression of mRNA transcripts encoding these adhesion molecules during the terminal differentiation of C2 mouse myoblasts. The accumulation of mRNA transcripts encoding N-cadherin, N-CAM, fibronectin, alpha 5-integrin, and beta 1-integrin subunits was developmentally, but not coordinately, regulated. N-cadherin and integrin subunit expression was maximal in prefusion myoblasts and declined thereafter. In contrast, N-CAM mRNA levels were low in prefusion myoblasts, and increased coincident with the onset of terminal differentiation. Fibronectin mRNA levels were also low in myoblasts, and they did not increase until after cell fusion had occurred. The results indicate that despite their lack of coordinate regulation maximal levels of mRNA transcripts encoding adhesion molecules are present at a stage which corresponds to the peak of the active phase of myoblast fusion.


Clonal analysis of cardiac morphogenesis in the chicken embryo using a replication-defective retrovirus. III: Polyclonal origin of adjacent ventricular myocytes. Replication-incompetent variants of the avian spleen necrosis virus (SNV) encoding cytoplasmic or nuclear-directed beta-galactosidase (beta-gal) have been used to trace the clonal growth of myocytes during left ventricular free-wall formation. Tubular-stage hearts were infected with a mixed suspension of both retroviruses and, after hatching, the progeny of marked cells in the ventricular wall were examined by X-gal histochemistry. When a small number of virions was introduced individual blue patches contained myocytes with only one label type (cytoplasmic or nuclear). These results confirmed our previous conclusion that each cluster or patch represents a single clone (Mikawa et al., 1992, Dev. Dynamics, 193:11-23). Each of these clones formed a clone-shaped patch which often extended through the entire thickness of the ventricular myocardium, but typically each patch was heterogeneous, containing a mixture of labeled and unlabeled cells. We then asked whether the two populations of myocytes in each patch were clonally related or generated from more than one progenitor. When hearts were infected with high titer viral suspensions many patches were observed in which cytoplasmic-tagged myocytes were intermingled with nuclear-tagged myocytes. Thus, the cone-shaped myocyte patches in the ventricular wall are polyclones derived from separate progenitors in the precardiac mesoderm. This finding led us to examine the separation of clonally related ventricular myocytes in the developing hearts. Embryos were infected with retroviral suspensions at varying stages of development and the resulting colonies examined after hatching.(ABSTRACT TRUNCATED AT 250 WORDS)


Comparison of Hensen's node and retinoic acid in secondary axis induction in the early chick embryo. Retinoic acid (RA) and Hensen's node, the organizer center in the chick embryo, have been shown to have polarizing activity when applied or grafted into the chick limb bud. Here we investigate and compare the effects of RA and grafted Hensen's node on the early chick embryo. Anion exchange beads soaked with RA at concentrations ranging from 5 to 100 ng/ml and implanted on the anterior side or on the left side of the host anteroposterior axis of a stage 4 chick embryo in ovo have the ability to induce secondary axis formation, while beads soaked with RA of the same concentration and implanted on the right side or on the posterior side of the host axis are unable to induce the secondary axis. All of the induced axes contain trunk-tail structures. Hensen's node from quail embryos implanted into the early chick blastoderm could also cause the formation of secondary axes in addition to self-differentiation of the graft into a secondary axis. Both RA and grafted Hensen's node caused the inhibition of forebrain development with an increase in hindbrain development and the host heart to loop in an abnormal direction. The results support the hypothesis that Hensen's node is a source of RA which is involved in early embryogenesis. Alternatively, RA might stimulate the formation of Hensen's nodal properties in adjacent tissue.


Optic chiasm and infundibular decussation sites in the developing rat diencephalon are defined by glial raphes expressing p35 (lipocortin 1, annexin I). p35, a Ca(++)-phospholipid-binding protein that serves as a substrate for the EGF receptor tyrosine kinase, is expressed by primitive glial ependymal cells to define a raphe occupying the ventral midline in the spinal cord and hindbrain of rat embryos (McKanna and Cohen, Science 243:1477-1479, 1989). p35 appears transiently in the median one-third (80 microns) of the floor plate at precisely the time and place where axons cross to form the ventral commissure. We postulated that if p35 is involved with commissure development, homologous p35 raphes might be found at decussation sites rostral to the floor plate, including the optic chiasm. The present report describes two developmentally regulated p35 raphes in the diencephalon. One raphe is present for 2-3 days at the rostral lip of the nascent infundibulum, the reported decussation site of axons running from the supraoptic nucleus to the neurohypophysis; the other raphe appears in the rostral two-thirds of the optic chiasm, the site traversed by the optic axons. p35 is never expressed in the caudal one-third of the chiasm that accommodates non-retinal axons. To the best of our knowledge, this is the first identification of a specific marker for the retinal component of the optic chiasm. Because the p35 is gone by embryonic day 18.5, it is absent during final stages of chiasm formation when axons from the temporal retina decussate. Thus, p35 also may contribute to the "barrier" perceived by fibers that remain ipsilateral. Our data suggest that the p35 raphe contributes to the midline's role in commisure morphogenesis. Putative lipocortin activities including regulating PLA2, eicosanoids, or intracellular Ca++ could be involved in altering cue specificity as decussating axon growth cones traverse the p35 compartment.


Expression of collagenase and IL-1 alpha in developing rat hearts. During development, extracellular matrix (ECM) molecules are thought to play a major role in regulating the formation of the heart. The change in the heart from a simple tube to a complex, four-chambered organ requires the modification of both the cellular components as well as the surrounding ECM. Matrix metalloproteinases (MMP), which include collagenases, are enzymes present in the ECM that have the potential to modify the existing ECM during the development of the heart. Using both monoclonal and polyclonal antisera against collagenase, specific temporal and spatial patterns have been documented during critical periods of heart development. The cytokine interleukin 1 alpha (IL-1 alpha), a potent inducer of the MMP expression, was also shown to have a similar staining pattern in the developing heart. The monoclonal anti-rat collagenase (Mab) intensely stained the surfaces of the myocytes in the trabeculae and the ventricular and atrial walls of the 11.5 or 12.5 embryonic day (ED) rat hearts. In contrast, the polyclonal anti-human collagenase (Pab) stained not only the cardiomyocytes but also the hypertrophic endocardial cells. Pab appeared to stain the leading edge of the mesenchymal cells that migrate into the cardiac jelly of the 11.5 or 12.5 ED hearts. Immunohistochemical staining showed IL-1 alpha on the endocardial endothelium and the surface of cardiomyocytes near the cardiac jelly just before or coincident with the appearance of migrating cells. IL-1 alpha was detected on the endocardial endothelium, cardiomyocytes in the trabeculae, and the ventricular and atrial walls, as well as in the myocardial basement membrane of the truncal or atrioventricular region. However, no staining could be detected on the migrating cells in the cardiac cushions. These results indicate the presence of collagenase and IL-1 alpha on the surface of cardiomyocytes and mesenchymal cells at times when the heart is undergoing acute remodeling during septation and trabeculation. These data suggest a role for collagenase/cytokine interaction in tissue remodeling during critical stages of cardiac embryogenesis where modification of the ECM is essential to cardiac morphogenesis.


Study of the behaviour of the mumps virus in different cell culture systems. The Enders reference strain of the mumps virus was cultivated on various systems (chicken embryonated eggs, chicken embryo fibroblast cultures, Vero cell cultures) to develop a model of preparing and controlling the production of the mumps vaccine in Romania. The results have proved that the monolayer of chicken embryo fibroblast is a suitable cell culture system for the multiplication of the mumps virus, and on the Vero cells the virus can be quantified just like the live measles vaccine.


Investigations concerning the possibility to replace the Schick test by the passive hemagglutination reaction for evaluating the diphtheria immunity level in population. The evaluation of Schick test results as compared to the results obtained by determination of diphtheria antitoxin concentration in blood using the neutralizing test on rabbit--Jensen on the one hand and to those obtained by the passive hemagglutination reaction on the other has revealed some discrepancies against the two tests: 7.09% (4.08% pseudo-protected, 3.01 pseudo-susceptible) as against TN-Jensen and 8.6% (5.59% pseudo-protected and 3.01% pseudo-susceptible) as against the passive hemagglutination reaction results. The reported discrepancies were due to the fact that the results of Schick test did not correlate perfectly with the amount of circulating antitoxin, the diphtheria immunity level was wrongly indicated by the Schick test. The passive hemagglutination reaction has proved to be adequate to mass-screening investigations being capable to replace Schick test.


Hemolytic properties of some Aeromonas strains. Considering the possible correlation between hemolytic and enterotoxigenic properties of Aeromonas strains mentioned in the literature, in the present work we studied the practical value of the hemolysis tests in the diagnosis of Aeromonas strains by using comparatively the hemolysis tube tests (with goat and sheep erythrocytes suspensions) as well as the technique on blood agar in aerobic conditions. There were studied comparatively 230 Aeromonas strains (different species: A. hydrophila, A. sobriae, A. caviae, A. veronii, A. salmonicida) isolated from different sources (meat products, fish of fresh waters, mussels, sea water, pipe water, diarrhoeal disease and animal faeces). The comparison among the used tests for proving the hemolytic activity of Aeromonas strains showed that: 61.89% and 56.51% from the total number of Aeromonas strains were hemolytic in the tube tests with 1% goat and 1% sheep erythrocytes suspensions respectively after 48 hrs followed by 51.72% and 48.25% strains by the 5% goat and 5% sheep blood agar plates respectively in anaerobiosis and after 48 hrs incubation at 37 degrees C. The highest incidence of hemolysin presence was pointed out in motile Aeromonas strains (A. hydrophila--60.11% and A. sobria--56.52%). Our results showing a high frequency of hemolytic activity among Aeromonas strains isolated especially in meat products are suggesting a possible correlation between the pathogenic potential and the hemolytic activity and are pleading for introduction of this test in the diagnosis of Aeromonas species.


Evaluating the minimum active concentrations of penicillin G and V and of retard forms in clinical conditions. Following several signals indicating the inefficiency of the clinical treatment with various penicillin preparations in some cases, we decided to study the seric penicillin concentrations in the patients hospitalized in the "V. Babes" Hospital of Infectious Diseases, after administration of the various Romanian made forms of penicillin currently used in the therapy of streptococcal infections and in the prophylaxis of the sequelae of these infections. The data obtained on groups exceeding 30 persons by using two methods of determining the penicillin concentrations the dilutions and the diffusimetric methods revealed protective penicillin seric levels satisfactory for penicillin G and Efitard, according to the present treatment schemes. After 5 days from Moldamin administration only 45.4% of children and 43.3% of adults were found to have satisfactory penicillin concentrations. The administration of penicillin V reaches active penicillin concentrations in terms of the dose administered. The paper points out only one of the causes which together with others (such as beta-lactamase production and tolerance), contribute to the unsuccessful treatment with various forms of penicillin.


Group B streptococci implicated in the etiology of acute pneumopathies in neonates. 111 neonates with acute respiratory insufficiency and/or severe pneumopathy (pneumonia, bronchopneumonia) occurring in the first 10 hours after birth were investigated during January 1990-September 1991 for the group B streptococci presence. 430 samples from these neonate cases: throat and nose exudates, necroptic material from died neonates (pulmonary, blood, bronchial secretion) and their mothers (lochia, vaginal and uterine secretion) were examined. 33% of the neonates and their mothers were positive with the same GBS. 65.30% from the GBS strains belonged to serotype Ia and 34.70% to Ib. 24.48% from the GBS strains were resistant to Penicillin G by diffusimetric method with a corresponding MIC of 0.1 UP/ml (22.12%) and 0.5 UP/ml (2.65%).


Sensitivity to penicillin of S. pneumoniae strains isolated from various pathological conditions. 297 S. pneumoniae strains isolated from patients with different pneumococcal infections during 1990-1991 were tested for the sensitivity to antibiotics--by the diffusimetric method to 9 antibiotics (Penicillin (6 micrograms), Ampicillin (10 micrograms), Erythromycin (15 micrograms), Oxacyllin (5 micrograms), Streptomycin (50 micrograms), Tetracycline (50 micrograms), Chloramphenicol (50 micrograms), Rifampicin (6 micrograms), and Kanamycin (30 micrograms) and by MIC determination to Penicillin and Erythromycin. 30% of S. pneumoniae strains were resistant to Penicillin and Erythromycin corresponding to a MIC > or = 1 UP/ml or > or = 1 microgram E/ml. The most active antibiotics were Chloramphenicol and Rifampicin and the less active: Tetracycline, Kanamycin and Streptomycin. A relation between the origin and serotype of the strains and the sensitivity to antibiotics was revealed. The strains from the throat, conjunctive and otic secretions and belonging to serotypes 19, 6 and 14 showed the highest levels of resistance to all antibiotics. A good correlation between MIC and diffusion method results was observed to Erythromycin and no correlation to Penicillin. In this last case the results of MIC determination to Penicillin were better correlated with the results of inhibitions diameters to Oxacyllin, method which have to be recommended.


Simultaneous recording of urethral pressure and cross sectional area profile. A new technique for simultaneously recording urethral closure pressure and cross sectional area profile is described. The cross sectional area of the urethra could be measured using the field gradient principle. The procedure was performed readily in male patients and the recordings can be graphically presented during measurement. A high degree of reproducibility using this method was obtained. Typical recordings of control subjects was well as patients with pathology were obtained using this technique. The results of this study show that the maximum urethral closure pressure obtained was 60.8 + 3.4 mmHg and functional profile length, 4.8 + 0.4 cm. Internal meatus cross sectional area was 0.67 + 0.04 cm2 and the minimum urethral cross sectional area was 0.12 + 0.02 cm2. This method has been shown to be readily applicable in measuring urethral closure pressure in males and can potentially be used as a tool of comparing the functional parameters of pressure with the anatomical value of diameter.


Study of strains agglutinable with enteroinvasive anti E. coli polyvalent sera isolated from ill and healthy subjects for optimizing the laboratory diagnosis of diarrhoea. 369 E. coli strains that are agglutinated in A or B anti E. coli enteroinvasive (EIEC) polyvalent sera and inducing no keratoconjunctivitis in the guinea pig's eye have been selected. The biochemical reactions of E. coli strains that can be agglutinated in A or B EIEC polyvalent sera occupy an intermediary position between those of Shigella genus and the classical features of Escherichia, i.e.; 87% are motile bacteria, 88%--ferment lactose, 81% form acid from sodium mucate and lysine is not decarboxylated by 87% of the strains. As for the antigenic behaviour with 15 out of the 16 sera employed, the strains under consideration have agglutinated by slide test in one or several sera, with varying intensities. None of the strains agglutinated on slide in the O164 antiserum. Subsequently, agglutinations have been performed in tubes only with the strains that agglutinated on the slide with a +3 or +4 intensity. Out of the 369 strains under consideration only 25 have agglutinated in the tube, all the false positive reactions representing 6.7%. The data presented lead to the conclusion that such strains that have been isolated from ill or healthy patients cannot be considered as pathogenic diarrhoea agents, the differentiation from the invasive strains (in the absence of the guinea pig for the pathogenicity test) cannot be made on the basis of the biochemical reactions, but only by means of a set of monovalent sera and by performing the tube agglutination tests.(ABSTRACT TRUNCATED AT 250 WORDS)


[Effect of AS-4370 on gastric motility in patients with diabetic autonomic neuropathy]. Delayed gastric emptying in diabetic patients occurs with progress of automatic neuropathy as one late complication. Delayed emptying is deeply correlated with poor glycemic control, due to imbalance between nutrients absorption and effect of exogenous insulin. AS-4370 is a newly developed prokinetic agent which has been reported to selectively activate motility of the upper gastrointestinal tract through enhancing acetylcholine release from nerve terminals within the enteric mural plexus. In this study, we evaluated the effect of AS-4370 on gastric motility in diabetic patients with autonomic neuropathy. Eight diabetic patients with autonomic neuropathy (3 males and 5 females) with mean age of 56 years old (range 29-66) participated to this study after giving written informed consent. Gastric motility was evaluated by gastric emptying and electrogastrography. Gastric emptying study was done using 99mTc-Tin colloid labeled omelet meal served with 2 slices of toast and 200 ml of milk. Electrogastrography was recorded from epigastric skin surface, for 30 minutes before and after meal each. AS-4370, 7.5 mg tid, was given for four weeks after basal recording of gastric motility studies. Following the 4-week treatment with AS-4370, gastric motility studies were repeated. For the motility studies after medication, drug was given 30 minutes before test meal. Gastric retention rate at 150 minutes in all patients were over 45% of upper limit of normal range in basal study with mean value of 69 +/- 5%, which decreased significantly to 52 +/- 5%, with AS-4370 treatment (p < 0.01). Gastric emptying speed, another parameter for gastric emptying also improved with medication.(ABSTRACT TRUNCATED AT 250 WORDS)


Spectrophotometric assay of the interaction of Plasmodium berghei infected erythrocyte lysates and neutral red. In order to reveal by absorption spectrophotometry the redox differences between the Plasmodium berghei infected erythrocyte lysates (MEL) and the healthy ones (HEL) we studied their interaction with the neutral red (NR) redox dye. The variation of the dye absorption intensity at 540 nm as a function of the hemoglobin content of the samples was attributed to the redox potential variation of the different hemoglobin aggregates formed in the samples containing different hemoglobin quantities. By short term treatment of the lysates with ascorbic acid in the presence of NR, great redox reactivity difference was proved between MEL and HEL as revealed by absorption and explained by the initial higher oxidation state of the Hb iron in MEL than in HEL.


Morphometric computerized analysis on the dentinal tubules and the collagen fibers in the dentine of human permanent teeth. A morphometric analysis has been performed on important components of human dentine using an image computerized analyzer. The dentinal tubule diameter and their area percentage were calculated. Moreover the area percentage of the collagen fibers in the dentinal matrix was measured. These parameters have been evaluated in different areas of the coronal and the radicular dentine in permanent teeth. Measurements have been performed on undecalcified and decalcified teeth and on teeth treated with enzymatic digestion to remove the organic non collagen matrix and to evidentiate the collagen fiber network. The values obtained in different areas of the tooth and in samples submitted to different treatments were evaluated by statistical analysis. Dentinal tubule diameter and area percentage significatively decrease from the inner to the peripheral dentine both in the undecalcified teeth as in the decalcified ones and in the samples undergone to enzymatic digestion. The collagen fiber percentage in the organic matrix is significatively lower in the mantle dentine.


[The effect of 2 bleaching agents on the enamel surface. An in-vitro study]. We present a study "in vitro" of the effect of bleaching agents on dental surfaces using the "Walking bleaching technique". We found that hydrogen peroxide bleached more quickly than carbamide although, after a period of six weeks, the results were the same as far as whitening was concerned. In the scanning electron microscope we observed significantly different changes in each case. Carbamide caused a regular and uniform opening of the enamel prisms of the surface while hydrogen peroxide produced more severe superficial destruction with the appearance of patterning similar to the acid etching, and the presence of some crystalline areas emerging from the body of the prisms.


[The bone structure of the ramus of the mandible]. Osseous structure of the ramus of mandible (RM) is of a practical clinical significance. Osteosynthesis of fractured segments and the success of the sagittal or the horizontal ramus split osteotomy depends on the cortical bone disposition and thickness. After morphometric investigations of the RM, conducted on 70 mandibles of adults, consecutive horizontal and frontal sections were made. On these sections, the cortical layer was studied and the cortical bone thickness was measured at four previously marked points. In the regard of morphometry, the significant datum is that nearly half of all the cases is grouped round the mean value of any parameter. Cortical bone is continuous and its two main sheets are the buccal and the lingual cortical plates. In its entirety, the buccal cortical plate is thicker than the lingual. The thickness of both cortical plates increases in the direction from the coronoid process to the angle of mandible.


c-myc hypermutation in Burkitt's lymphoma. Translocation between the c-myc protooncogene and one of the three immunoglobulin loci is a cytogenetic hallmark of the B cell tumor, Burkitt's lymphoma. The resulting deregulation of c-myc expression is a critical step in tumorigenesis. The translocation breakpoint may lie within c-myc proper, in which case deregulation is due, in part, to dissociation of key 5' regulatory sequences from the protein-coding portions of the gene. Alternatively, the breakpoint may flank c-myc, leaving the gene grossly intact. In these latter cases, mutation, which may be extensive, is usually seen within c-myc, specifically at or near the same key regulatory sequences. The precise contribution of these mutations to c-myc deregulation is gradually being clarified. The mechanisms underlying c-myc mutations are not known. Hypermutation in c-myc may reflect the influence of the juxtaposed immunoglobulin gene, which is subject to hypermutation during an intermediate stage of normal B lymphoid development. This relationship, however, has not been firmly established.


NaOCl and EDTA irrigating solutions for endodontics: SEM findings. Premolars roots of humans were manually instrumented with K-type files and irrigated with different solutions to evaluate the rate of cleaning of endodontic surface. Root canals irrigated with 0.9% saline solution or H2O2 (10 volumes) showed the presence of predentin and amorphous smear layer. Thick smear layer was always present on endodontic walls rinsed with 5% solution of NaOCl. Specimens treated with 0.2% solution of EDTA showed partially clean dentinal tubules orifices and remnants of a thin smear layer. Occasional uninstrumented areas of the same roots presented smear layer remnants and predentin with calcified bacteria. The root canals irrigated with NaOCl and EDTA solutions alternated after each instrument showed at the dentin surface thick smear layer: only few dentinal tubules orifices were visible. Endodontic surface of root canals irrigated with NaOCl during instrumentation and finally rinsed with EDTA solutions showed the most homogeneous ultrastructural pictures: partially clean dentinal orifices were detectable in the whole canals.


[Light and electron microscopy observations on different tissues of a complex odontoma]. Several aggregates of dental tissues constitute the complex odontoma. They are almost completely covered by a layer of prismatic enamel. By the observations at transmission and scanning electron microscope enamel present the characteristics of a not fully developed tissue. The organic component is still abundant and the enamel surface is covered by a cellular layer having the morphological features of the mature ameloblasts. Therefore this enamel results to be yet actively engaged in the maturation phase although the tissues of the odontoma have generally a limited developing time.


Human interleukin-9: a new cytokine in hematopoiesis. IL-9 was first identified in the mouse system as a T cell growth factor while human IL-9 was isolated based on its activity on a human myeloid leukemic cell line. The high sequence homology between mouse and human IL-9 plus the preliminary biological analysis have predicted a wide spectrum of in vitro biological activities for this cytokine. Despite the high sequence homology between mouse and human IL-9, many biological activities, however, have not been demonstrated in both species. In vivo studies therefore become essential to understand the physiological role of IL-9. Increasing evidence have also suggested the possible role of IL-9 in the pathogenesis of HD and LCAL. Detailed analysis of IL-9 expression, autocrine growth and clinical outcome of HD patients will be required to make any speculation on the role of this cytokine in the disease states.


[Enamel pearls: the ultrastructural aspects and morphogenesis hypotheses]. Five enamel pearls were examined by scanning electron microscope. Some external pearls presented a bare enamel surface, others were covered by a thin layer of cementum. The enamel pearls showed irregular dystrophic enamel surface or a thin layer without structure, rows of irregular Tome's processes pits, enamel caps and focal holes. Some enamel pearls revealed little areas of enamel surface resorption like-resorbing lacunae with, in some cases, areas of tissue repair.


[The effect of the magnification ratio on 2 methods of computerized image analysis in scanning electron microscopy]. The aim of this study is to compare two computerized analysis methods of SEM dentinal walls views. The first technic is an indirect one using a primary marking and a secondary measurement. The second one is a direct threshold analyse, using the grey levels selection. The influence of the magnification is analysed in a second part. The first technic is considered as a reference one. The results show that the threshold measurement technic is only reliable at a high magnification. A direct surface measurement cannot be used at a low magnification. A pretreatment could enhance the ability of a computer to measure the areas covered with smear-layer or organic debris on SEM micrographs.


The interleukin 2 receptor in the hypereosinophilic syndrome. The hypereosinophilic syndrome (HES) has been previously described as a clinicobiological entity characterized by a blood eosinophil count of over 1.5 x 10(9)/L of unknown cause associated with several clinical complications. In reality, HES is a heterogeneous group of diseases with variable and unpredictable progress in visceral lesions, thought to be related to the deleterious effects of tissue eosinophil infiltration. Various criteria for discrimination between benign and severe forms of HES have been described. These previous retrospective clinical investigations, using biological and clinical markers, have defined different stages of HES. It appears more relevant, however, to consider elements of disease activity by studying mechanisms of induction of persistent hypereosinophilia. The T-cell dependence of blood eosinophilia has led us to evaluate various markers of T-cell activation in particular. In the present review, we report previous results and perspectives suggested by the study of the interleukin 2 receptor in HES.


Transient abnormal myelopoiesis in Down's syndrome. Recent data have elucidated the pathogenesis of transient abnormal myelopoiesis (TAM) to a great extent. TAM is a monoclonal disorder which resolves spontaneously and the target cell in this disorder is a multipotent stem cell which is capable of differentiating into megakaryocytes. The pathogeneses of TAM/AMKL (acute megakaryoblastic leukemia) appears to be closely associated with abnormal quality and quantity of a gene located on chromosome 21. AMKL developing after the regression of TAM appears to come from the same clone as the TAM, which apparently experiences some kind of genetic alterations. It seems that the gene responsible for TAM will soon be cloned in the near future. However, the mechanism of spontaneous regression of TAM has as yet not been clarified. The expanding clone in the transient physiological immunodeficient state, during the perinatal period, might be eliminated with the maturation of more mature immunosurveillance. Alternatively, the TAM clone might be destined to undergo spontaneous death, which is called "programmed cell death" (apoptosis). The mechanism of this phenomenon awaits further elucidation.


The value of the bone marrow plasma cell cytoplasmic light chain ratio in differentiating between multiple myeloma and monoclonal gammopathy of undetermined significance. We compared the plasma cell light chain ratios in the bone marrows of 13 patients with multiple myeloma (MM), with those of 13 patients with monoclonal gammopathy of undetermined significance (MGUS). The mean light chain ratio in favour of the paraprotein isotype in the myeloma group was 51.83 (95% confidence limits (CL) 29.52-74.14), while in the MGUS group it was 5.30 (CL 2.07-8.52). The difference between the MGUS and MM groups was significant (p = 0.0005). Neither the bone marrow plasma cell count nor the paraprotein level were significantly correlated with the light chain ratio in either of these two groups. We found a cut-off ratio of 8 to be the most useful in differentiating between myeloma and MGUS. Only one patient with myeloma had a ratio below 8, and one MGUS patient had a ratio above this cut-off point. We conclude that determination of the bone marrow plasma cell light chain ratio is a simple and useful test in differentiating between myeloma and MGUS in difficult cases.


Pulmonary involvement as the major manifestation of chronic lymphocytic leukemia. Respiratory symptoms and abnormal findings on chest X-ray are frequently noted in patients with chronic lymphocytic leukemia (CLL). However, most of these represent pulmonary infections or mediastinal lymphadenopathy, and leukemic involvement of the lung is seldom diagnosed during life. In this report we describe three patients with non-progressive, responsive CLL who developed biopsy proven pulmonary infiltration with CLL. In one case, pulmonary involvement was the sole manifestation of recurrent disease and a second case had little disease elsewhere with minimal CLL in the blood at the time pulmonary involvement appeared. In all three cases, transbronchial biopsy and bronchoalveolar lavage performed during fibreoptic bronchoscopy provided adequate tissue for diagnosis. We conclude that CLL may involve the lung even in the presence of a low peripheral white blood cell count with responsive disease elsewhere, and can readily be diagnosed by transbronchial biopsy and bronchoalveolar lavage.


Polycythaemia vera following non-Hodgkin's lymphoma. A patient with non-Hodgkin's lymphoma (NHL) who developed polycythaemia vera (PV) is reported. Diffuse large-cell NHL was diagnosed and he was subsequently treated with combination chemotherapy including high dose cyclophosphamide and procarbazine. Four and a half years after chemotherapy splenomegaly developed, coincidently with the appearance of high Hb values, RBC and platelet counts. The screening tests for PV were consistent with this diagnosis, while the search for lymphoma activity was negative. To the best of our knowledge, the present case represents the first well-documented instance of PV following NHL.


Urinary bladder cancer following cyclophosphamide therapy for Hodgkin's disease. Urinary bladder cancers following prolonged cyclophosphamide therapy are being increasingly reported. We report a case of transitional cell carcinoma of the urinary bladder occurring 12 years after pulse intravenous therapy with cyclophosphamide for Hodgkin's disease. The mechanism of bladder carcinogenesis and the possible role of the uroprotector MESNA in preventing cyclophosphamide induced bladder cancer are discussed.


[The psychophysiological bases for the training of flight personnel for operations in contingency situations]. The paper deals with the psychological and physiological mechanisms characterizing the behavioral patterns of a pilot under emergency situations. In a regular flight, the most important psychic regulator is a target-image and predicting of an assigned task performance serves as the mechanism of its implementation. In non-routine situation, the regulation pattern is significantly changed. Under emergencies, there exist two levels of pilot responses: adaptive-defensive reactions which activate the automated acts and evaluation-decision-making ones which promote intellectual transformations of information to compare the conceptual and information models. In order that under emergency the pilot response be successful the specific mental skills are needed to promote the rate of abstraction and determination of key sings which are the basis for decision making. Experimental data on the reliability and adequacy of pilot response in real flight under simulated contingencies are presented. Typical disorders of cognitive psychic processes, behavioral disorganization under these conditions have been described. The components of psychophysiological adequacy of pilots and principles and means of its creation are discussed.


[The principles of constructing a data bank applicable to the problem of physiological standards in space medicine]. The norm is a more complex problem in physiology and in space medicine in particular. Definition of norm and health concepts is analyzed. Classification of functional states based on the adaptation and homeostasis concepts is proposed. Various approaches to determination of a norm are described. Three types of the norms which are of significance for space medicine, namely rest norm, reaction norm, adaptive norm are identified. The structure of data bank containing information on the standards is described. The central data bank structure is considered.


[Altitude stress and the training of cosmonauts]. An ergonomic concept of the development of altitude stress is presented. According to this concept, the altitude stress as a psychophysiological response of man to the risk of falling from altitude is determined by the number and placement with respect to man position of antifalling contact supports and those being non-contact, i.e., potential supports capable of preventing the fall when changed a situation with the structures. The analysis indicated that this concept is also applicable for describing the extravehicular locomotion of cosmonauts with the use of the supports on the basis of which the preliminary ground preparation of cosmonauts using altitude-spatial trainer becomes possible.


[The methodology for clinical trials of radiation-protective agents intended for the conditions of manned space flights]. The methodology of clinical testing of antiradiation measures to be used in manned space missions is proposed and discussed. The tests should be based on: an individual tolerance to a recommended medicinal form of the drug; the tolerances to the effects of motion sickness, prolonged transverse and longitudinal accelerations; graded physical exercise; evaluation of performance (operator activity).


[Adaptation to anthropogenic chemical compounds in human applied ecology]. The literature data pointing to the fact that man can actively adapt to the effect of chemical substances at doses not exceeding hygienic norms are presented. However using some medical measures it is possible to increase to some extent the body resistance to the effect of chemical impurities of the environment and thus delaying for some time the development of disadaptation disorders induced by these impurities.


[The cabin microflora of manned space vehicles and the problem of the biological destruction of the construction materials used in them]. It is found that during operation of manned space objects there can occur the microbial growth on the structural materials of cabin interior and equipment which results in their biological destruction. The biodestructing processes can involve single parts of cabin interior, trainer linkages, connectors, cables, illuminators. Ninety four microbial species have been identified in spacecraft environment. The bacterial flora was presented by 19 genera pertaining to 11 families and involved 58 species. The mold and yeast-like fungi were presented by 12 genera and consisted of 36 species. The significant quantities of isolated microbial species were classified as potentially pathogenic microorganisms; many of them are known as biodestructers of polymeric materials. The isolated microbial cultures in space missions have become the basis of strain bank intended for further study and practical use in validating the construction materials.


[The cardiovascular system function of the cosmonauts of the 6th prime expedition on the Mir station]. Medical results of inflight cardiovascular measurements made in the 6th prime crew at rest and during provocative graded bicycle exercises and LBNP tests are presented. The methods of electrocardiography, tetrapolar rheography, kinetocardiography, arteriovenous pulsography and tacho-oscillography have been used to examine the cosmonauts. Together with regular cardiovascular changes typical of microgravity effect the individual peculiarities of developing the control mechanisms are noted. Thus in one of the cosmonauts a number of symptoms responsible for a decreased venous return. As a whole the functional state of the cardiovascular system on all flight stages was characterized by a sufficiently high physiological level.


[The ultrasonic location of gas bubbles in the human bloodstream during work in a spacesuit]. The results of testing a procedure of ultrasonic location of the gas bubbles (GB) in man during space suit operations to simulate an extravehicular activity (EVA) are presented. Doppler echotachocardiograph "Rhythm" operating at ultrasonic frequency of 1.76 mHz was used as a GB detector. The device "Rhythm" integrated with a special flat transducer of 23 mm in diameter and 4 mm in width was positioned on the subject chest above the pulmonary artery projection. During 4-6 hour human operations while wearing pressurized (276-290 mm Hg) space suit, in 7 of 12 tests performed a stable and qualitative signal of the arterial blood flow in the lungs was recorded. In case of an unstable signal, in order that its quality be improved the posture of test subject was changed and the signal was recorded during a short-term expired breath-holding. Cardiac GB formation was noted during 6 tests in 2 subjects. The first GBs appeared at the 30th, 33rd, 70th, 111th, 114th and 180th minute after producing an operating reduced pressure in space suit. The time of GB detection on the average was 89.7 min. The maximum intensity of GB signal was 3-4 scores on Spencer's scale, but altitude decompression sickness symptoms did not develop in the test subjects. The risk of developing the altitude decompression sickness as opposed to the results of control experiments without use of space suit is discussed.


[Morphological research on the liver structures of experimental animals under the action of infrasound]. Data of the experimental investigations of the effects of infrasound (frequencies: 2, 4, 8, and 16 Hz; acoustic pressure: 90, 100, 110, 120, 130, and 140 dB) on hepatic structures of laboratory animals are presented. Single exposure to infrasound with any of the given parameters induces no changes in hepatic structures. Such elements of the liver as hepatocytes and small vessels turned out to be most sensitive to 3 h./d infrasonic exposure during 5 to 40 days. Alterations in the liver were local by nature and manifested by ischemic areas with hepatocyte morphologic and histochemical changes. Hepatocyte changes after 5 to 40-day infrasonic exposure showed increased functional activity. More durable exposures (25 and 40 days) yielded irreversible changes in hepatocytes. These results indicate that 8 and 16 Hz infrasound produces more pronounced histological, histochemical and ultrastructural alterations than 2 and 4 Hz. These data suggest that the above frequencies can indirectly impair cell organoids and nuclear chromatin, and affect the hepatocyte function.


[The asthenic syndrome and mental work capacity]. Investigation of the development of the asthenic syndrome during neuropsychic overload and its impact on dynamics of psychic performance in operators is of great interest to many specialists engaged in clinical and applied medicine. The paper presents results of clinicopsychologic and clinico-neurophysiologic studies of 105 persons in intellectual and operational professions who were treated in Moscow Clinic of neurosis in connection with the asthenic syndrome caused by professional overload or emotional stress. The obtained data showed that development of the asthenic syndrome had certain stages depending on duration and severity of emotional, vegetative, dissomnic and other psychosomatic disorders. It was shown that, being one of the key integral indices of human adaptability, psychic performance retained on sufficiently high level for a long period during asthenic syndrome.


[The protein composition of the blood plasma of cosmonauts after prolonged orbital flights]. The method of two-dimensional electrophoresis was used to determine the protein composition of blood plasma in the cosmonauts stayed on board Mir station over 166 and 366 days. Separation pattern of plasma protein observed next day after landing had several typical differences from the standard pattern of plasmatic proteins found during a preflight examination and on the 7th postflight day. The changes were an increase in intensity of specific protein spots as well as in occurrence of unusual protein spots.


[Video display terminals: their electromagnetic safety]. Widespread application of video terminals (VT) has led to accumulating some unambiguous data on harmful effect of electromagnetic and electrical fields on the health. An analysis indicates that the level of electromagnetic and electrical fields is below permissible limits. IRPA/INIC concludes that the electromagnetic effects on VT users health are unharmful. However, the problem of VT use is an integrated one and involves not only radiation but ergonomic aspects and in this context requires further studies.


[The righting reaction in free fall in labyrinthectomized rats after a flight on the Kosmos-936 biosatellite]. The paper presents the experimental data on a turning over reaction in labyrinthectomized rats after 18.5 day flight on Cosmos 936. On Earth, the labyrinthectomized rats are found to exhibit with time an activation of gaze fixation reflex (GFR) which substitutes for labyrinth function when executed a turning over reaction. In microgravity, in the labyrinthectomized rats there is no activation of GFR and postflight turning over reaction is absent in the rats.


Recent pathogenic concepts of primary open angle glaucoma and soft glaucoma (POAG). Loss of mainly field in POAG is due to Schnabel's cavernous atrophy of the axons at their scleral passage, built up by aligned stacks of glial collagen (IV and I) tissue plates, which contain capillaries. Organisation, size and structural surroundings encircle isolated or grouped axons not evenly distributed. The stereotype pattern of fieldloss linked to a characteristic optic nerve lesion suggests POAG be considered a N.O. disease as raised IOP may be only one of the risk factors. Are cavernous atrophy (loss of field) and the trabecular spaces narrowing (rise of IOP) both the result of the same pathogenic mechanism acting on the collagen fiber (IV and I) of the scleral coat, resp. lamina cribrosa (posterior segment) and trabecular meshwork (anterior segment)? If so, IOP-rise only worsens the condition as will do a hemodynamic energy crisis at the level of the axons. Interactive dominance at any of both levels may explain unexpected discrepancies between IOP and fieldloss. Considering "to treat or not to treat" accurate field loss predictability testing, urgent research topic, are discussed.


Diagnosis and long term follow up of glaucomatous visual field defects with automated perimetry. Automated perimetry has added a very important standardized observation post to the clinician's perspective on glaucoma. This paper reviews the major characteristics of field loss in glaucoma as detected with automated perimetry. Practical problem cases are used to illustrate and emphasize specific areas of interpretation difficulties.