The low-titer antibodies against HTLV-I were determined using HTLV-MA method and the age distribution of the antibody titers was clarified in HTLV-I carriers. Twelve hundred and three serum samples were obtained from the people living in the southwestern part of Miyazaki prefecture, where HTLV-I infection is endemic. They were examined by indirect immunofluorescent method using membrane antigen of alive HUT102 cells. One hundred and ninety-three sera were found to be positive for HTLV-I, but the antibody titers of 24 samples were as low as 1: 4. About 50% of the carriers under the age of 20 showed a low titer of the antibody, whereas, when the carriers were old, the rate of low-titer antibody was low. The rate of high titer (≥1: 160) antibody was nil in the carriers under the age of 30, and it increased as the age advanced. The geometric mean titer of the antibodies was significantly higher in the carriers above the age of 30 than that of those under 30.
Sera from HTLV-I carriers were studied with immunoprecipitation using [35S]-cysteine labelled HUT 102 cell lysate as an antigen and were analyzed further by SDS-polyacrylamide gel electrophoresis. The viral proteins which specifically reacted with the the sera from carriers were mainly gp61, p55, gp45, p42, p24 and p19. Analyzing the thickness of bands on electropherograms, the intensity of antibody reaction to the above proteins was strongest in gp61 and in decreasing order of gp45, p55, p24 and p19. Secondly the incidence of detecting the antibodies to each viral protein was studied. In the lowtiter sera, the antibodies to envelope protein such as gp61 and gp45 were mainly detected, while the antibodies to core proteins such as p55, p24 and p19 were found in the high-titer sera. The antibody to gp61 was demonstrated in almost all the samples, and the thickness of this band was well correlated with the antibody titer of the sera. p42 is encoded by pX region of HTLV-I gene. The thickness of the band of anti-p42 antibody and the incidence of detecting this antibody had no relation to the antibody titer in contrast to the above proteins. This observation suggests that the mechanism and process of anti-p42 antibody production appears to be different from those of other proteins of this virus.
Egg yolk-Polymix B-Glycine-Salt (EPGS) agar was devised for the rapid isolation of Staphylococcus aureus from feces of patients with food poisoning. The medium consisted of 10g of mannitol, 3g of glycine, 5g of lithium chloride, 10mg of polymyxin B sulfate, 25mg of phenol red, 18g of agar, and 900 ml of distilled water. It was autoclaved at 121°C for 15 minutes. 100ml of a 30% egg yolk emulsion were added to the medium after it had cooled to 50°C. It was found that the presence of 0.3% glycine, 0.5% litium chloride and 0.001% polymyxin B sulfate in the medium showed a considerable inhibitory effect on Bacillus subtilis strains, while the medium had enhanced the growth of all S. aureus strains used in the experiment. When the EPGS agar was applied to feces of healthy people and patients with food posisoning, S. aureus was detected more effectively than by the Mannit Salt Egg Yolk (MSEY) agar. On the EPGS agar, colonies with egg yolk reaction could be counted as specifically S. aureus after an incubation time of 24 hrs, whereas the MSEY agar required 48 hrs to grow S. aureus from feces of patients with food poisoning. The EPGS agar may be useful for a routine laboratory test for detection of S. aureus in feces of patients with food poisoing.
The studies on combination therapy of norfloxacin (NFLX) and fosfomycin (FOM) were performed, and following results were obtained. 1. The fractional inhibitory concentration index (FTC index)≤0.5 between NFLX and FOM showed 76.5%(26/34) against P. aeruginosa isolated from respiratory patients for respiratory tract infection and 60.9%(14/23) against methicillin resistance S. aureus. The synegistic effect was confirmed in this in vitro study. 2. The FIC index derived from the combination therapy was 0.06 agaisnt one strain of P. aeruginosa shown MIC of 100μg/ml in NFLX and MIC of 600μg/ml in FOM. 3. Clinical effect was good in 15 cases (93.8%) and fair one case (6.2%). 4. Bacteriologically 24 strains out of 25 strains isolated from 16 patients were eradicated after the treatment; 8/9 in P.aeruginosa, 6/6 in H.influenzae, 6/6 in S.aureus, 1/1 in S.pneumoniae, 1/1 in S.pyogenes, 1/1 in K.pneumoniae and 1/1 in Flavobacterium sp. 5. No adverse effect and abnormal labolatory findings were observed in this study. It is suggested that the combination of NFLX and FOM is useful for the treatment of respiratory tract infections.
Ultrastructural changes in the endometrium of rabbits with intrauterine infection experimentally induced with Bacteroids fragilis GAI 0558 were studied by electron microscopy. Two days after inoculation, transmission electron microscopy revealed a disarranged appearance of the endometrial microvilli. Numerous macrophages and polymorphonuclear leukocytes phagocytizing bacteria were observed near the endometrial mucosa. In six or seven days after inoculation the endometrial cells were degenerated and necrotized, and the infecting bacteria were observed among destroyed epitherial cells. In ten days after inoculation, inflammatory changes developed up to the myometrial layer. Through transmission electron microscopic findings revealed no evidence of infecting bacteria in the tissue of the endometrium, fluorescent antibody staining showed presence of the bacteria. Scanning electron microscopy revealed the infecting bacteria adhering to the endometrial surface. The glycocalyx, a universal surface component of the bacteria, surrounded the bacterial cells and bound them together to form microcolonies and it also mediated bacterial adherence.
To save labor and make it easy to take an immediate countermeasure at the time of isolation of influenza virus, studies were made on a method to inoculate a sample directly into cells which had been stored in a frozen state. The following results were obtained. 1. When MDCK cells were stored at -80°C with DMSO as a low-temperature protecting agent, they showed a survival rate of more than 95% after storage for 6 months. 2. MDCK cells stored in a frozen state presented a high susceptibility to the standard strain of influenza virus. 3. When used for the isolation of virus from samples collected from patients, MDCK cells stored in a frozen state gave results not inferior to those obtained from monolayer culture cells. 4. It was possible to take an immediate countermeasure when cells stored in a frozen state were used for the isolation of virus. Furthermore, the use of these cells could make it easy to isolate influenza virus even at a laboratory which was lacking in the equipment for cell culture and in experienced workers.
For the rapid diagnosis of respiratory infections caused by Pseudomonas aeruginosa, we developed IFA-staining of sputum for Pseudomonas aeruginosa using serogroup-specific monoclonal antibody (MEIASSEI RYOKUNOUKIN, MEIJI, JAPAN). IFA-titers of each serogroup-specific antibodies were distributed from 1: 4 to 1: 200 dillution, and minimal concentration of bacteria detectable by IFA-staining was 106 CFU/ml in the sputum. We performed IFA-staining on 131 smears of the sputa presented to the Clinical Laboratory of Nagasaki University. In 36 specimens from which more than 106 CFU/ml of Pseudomonas aeruginosa were isolated, 27 specimens (75%) were judged as IFA-positive, and in 18 specimens with more than 107 CFU/ml of Pseudomonas aeruginosa, 15 (83%) were IFA-positive. Furthermore, there was a distinct correlation between number of fluorescing bacilli (microscopic findings of IFA-staining) and quantity of Pseudomonas aeruginosa isolated from the sputum. In the 86 sputa with a negative culture for Pseudomonas aeruginoas, 82 (95%) were judged as IFA-negative. Using this method, we can directry identify Pseudomonas aeruginosa in the sputum, and according to these results, the IFA-staining of the sputum was considered to be a useful method for the rapid diagnosis of respiratory infections caused by Pseudomonas aeruginosa.
The annual examination for HBsAg and anti-HBs was carried out on personnel of Nagoya National Hospital during the 11 years from 1975 to 1985. In 1976 the frequency of natural positive turn of HBV markers was 3.3% in the medical staff and 2.5% in the non-medical staff, but in 1985 it showed a marked decrease to 1.0% in the medical staff and 0% in the non-medical staff. These results suggested an outcome of protective measures against hospital infection of HBV as well as a tendency to decrease HBV infection in social life. The frequency of natural positive turn of HBV markers in a year was highest in surgeons (1.9%) as compared to internists (0.9%), nurses (1.7%), laboratory technicians (1.0%) and non-medical staff (1.0%). Classifing by the disposition of nurses, it was significantly high in nurses working in the outpatient department (4.8%) as compared to the operating room (2.9%), maternity floor (2.2%), surgical floors (1.2%) and floors of internal medicine (1.1%). These results suggested the highest risk of HBV infection in the outpatient department.
2298 strains of group A streptococci isolated from patients with scarlet fever or pharyngitis at an outpatient clinic in Matsuyama City, during the 10-year period from 1975 to 1984, were examined by means of T typing, M typing, Opacity Factor (OF) test and by measuring MIC of PC G, AMPC, CEX, TC and EM. The results were as follows: 1) The T type number did not necessarily correspond to the same M type number. The results of T typing did not correspond to the results of M typing in 5% of T type 1 and 4 strains, and in 20% of T type 12 strains. 2) 30% of the “T type 12, M untypable” strains were OF test positive, suggesting that they belong to an M type other than 12. 3) Immune serum against Matsuyama 2166 strain (T type 12-28 complex, M untypable) had a specific M antibody to Matsuyama 2166 strain but reacted neither to M 12 nor to M 28 strain. 4) All of the isolates were sensitive to PC G, AMPC and CEX. T type 4 strains were resistant to TC and T type 12 strains were resistant to TC or both TC and EM. 5) M type 12 strains were resistant to EM until 1982 when the isolation rate of M 12 strains was the least, and were replaced from 1983 by EM sensitive M 12 strains, which have been increasingly isolated thereafter. It was concluded from these results that the type-prevalence of Group A streptococci based on the T typing method should be carefully reappraised by means of additional tests, such as the opacity factor test, and preferably by means of M typing.
In this study, we have examined the interaction between Legionella pneumophila serogroup 1 (80-045 strain), and human polymorphonuclear leukocytes (PMNs) and monocytes in vitro. With complement alone or antibody alone, few PMN or monocytes ingested L. pneumophila. Effective phagocytosis of PMNs and monocytes against L. pneumophila was observed only in the presence of both antibody and complement. However, L. pneumophila was not killed by PMN and monocytes, and these bacteria, even when incubated with antibody and complement, were only partially susceptible to killing of PMNs. Virulent guinea pig-through L. pneumophila (virulent L. p.) were more resistant than avirulent agar-grown L. pneumophila (avirulent L. p). Superoxide production from PMN and monocytes, which was observed when these cells phagocytosed E. coli, was not detected in phagocytosis of L. pneumophila. In the case of virulent L. p., superoxide production from phagocytic cells could not be found even with increased ratios of bacteria to cells and addition of antibody and in avirulent L. p., a little increase of superoxide production was observed only when the ratio of bacteria to cells was the highest. By electron microscopy of PMN ingested virulent L. p., avirulent L. p. or E. coli in the presence of both antibody and complement, all bacteria were found in the phagosome, and none were found free in the cytoplasm. The volume of the phagosome containing E. coli or avirulent L. p. was often large relative to the volume of the bacteria within it. These phagosomes frequently had more than one organism and were closely surrounded by multivesicular types of lysosomes. On the other hand, phagosomes containing virulent L. p. were not large and nearly all of these phagosomes had a single bacterium. Around these phagosomes, mitochondria and various types of lysosomes were observed. The findings which suggested fusion of lysosomes with phagosomes were observed often in E. coli, less often in avirulent L. p. and very rarely in virulent L. p. As stated above, L. pneumophila has some escape mechanisms from the killing system of the phagocytes and the details of these mechanisms should be evaluated in the future.
Anaerobic curved rods were isolated from a vaginal specimen of a patient suffered from Candida vaginitis. She was a 24 years old and had a previous history of using an intrauterine device. At 23rd June 1986, she visited and complained the increase of vaginal discharges. From the discharges, Candida albicans and Gardnerlla vaginalis were isolated. At 19th September, she visited again for the check-up after cystitis. From a vaginal specimen, anaerobic motile curved rods, which were identified as Mobiliuncus curtisii subsp. holmesii, were isolated with C. albicans.