Although it has been well established that Kanagawa phenomenon-positive Vibrio parahaemoly-ticus is a human enteropathogen, the Kanagawa phenomenon-negative one has been considered to be probably not pathogenic. We have found, howerver, an outbreak of gastroenteritis due to Kanagawa phenomenon-negtative V. parahaemolyticus which produces a new toxin (Vp-TRH) resmbling to Vp-TDH, a responsible toxin of Kanagawa phenomenon. In this study, we developed monoclonal antibodies against Vp-TRH which were used for development of an enzyme-linked immunosorbent assay (ELISA) for specifically detecting Vp-TRH. The ELISA was applied for analysis of production of Vp-TRH by various isolates of V. parahaemolyticus and we found that Vp-TRH-producing strains were derived mostly from human diarrheal stool, and not from the environment or sea foods. The results of the rabbit ileal loop test showed that Vp-TRH-producing (Kanagawa phenomenon-negative) strains, as well as Vp-TDH-producing (Kanagawa phenomenon-positive) strains could induce fluid accumulation. These results indicate the possibility that Vp-TRH-producing Kanagawa phenomenon-negative V. parahaemolyticus is a human enteropathogen.
It has been well known that the transmission of HB virus is associated with mass inoculation and other medical procedures. In order to make clear the relation between HBV infection and the mass survey for filariasis on which sera of the examinees were drawn from their earlobes for the detecting of microfilaria with inadequately sterilized instruments, a epidemiological study was conducted in Miyako district, Okinawa prefecture, Japan, in which both HBV infection and filariasis were prevalent. A total of 2, 231 inhabitants were investigated for HBV seromakers. The over all positive rate of HBs antigen was 6.3% with the highest rate of 15.5% in the group of age 25 to 29. On the other hand, the prevalence rate of HBs antibody continued to increase along with age group, and remained to be over 60% after 30 years of age. The positive rates of both HBs antigen and antibody in the subject district were higher than those in other place of Okinawa prefecture or Japan. Comparing yearly attack rates of HBV chronologically, which were calculated from the agespecific HBV exposure rates in the district, the annual incidence of HBV infeciton was 6.1%(%/year) between 1954 to 1958, 5.2 (%/year) between 1959 to 1963, and then 4.4 (%/year) in the period of 1964 to 1968 in which the proportion of examinees for the survey of filariasis was the highest. In conclusion, the mass survery for filariasis was not the main factor contributing to the high prevalence of HBV infection in Miyako district.
The effects of somatic components of Lactobacillus casei (L. casei) were studied on cell growth in vitro. L. casei was able to suppress the growht of MT-2, MT-4 cells from adult T-cell leukemia, Molt-4 cells from acute lymphoblastic leukemia and U-937 cells from promonocytic leukemia. This effect was obviously different from the cytotoxicity of Vinbrastin, an anti-cancer drug. Flow cytometric experiments employing BrdU-anti BrdU antibody demonstrated an increase of cells in G1+GO phases (pre-DNA synthesis phases) by the treatment of L. casei, therefore L. casei maybe acts as a lowgrade inhibitor of the protein synthesis. PC-treated L. casei had no more inhibition on cell growth than the non-treated one.
Campylobacter jejuni strains isolated from gastroenteritis at 4 general hospitals of Tokyo Metropolitan during the period from 1981 to 1987 were serotype according to the slide agglutination test (TCK system) developed by the Tokyo Metropolitan Research Laboratory of Public Health. Two thousand four hundred seventy-nine strains isolated from sporadic cases among infants and children, 1, 962 (78.5%) were typed by 33 typing sera numbered TCK 1 through TCK 33 and leaving 537 strains (21.5%) untypable. Out of the typable strains, 1.643 strains reacted with only single serum, while 319 strains reacted with 2 or more antisera. The most common serogroups included TCK 21, 20, 7, 1, 4, 23, 24, 10, 30 and 12. Out of the 1, 250 strains isolated from sporadic cases among adults, 974 strains (77.9%) were typed and 276 strains were untypable. The most common serogroups were smiliar to those of infants and children. Serogroups TCK 1, 7, 4 and 21 were consistently the common serogroups every year during the 7 year study. Isolation frequency of serogroup TCK 30 have increased remarkably since 1986, while TCK 23, 14 and 9 have decreased.
During the period from May to September in 1984, an epidemic of aseptic meningitis occurred in Tajimi city, Gifu prefecture. Virological and serological investigations were performed. Cases involved ranged from 0 to 12 years of age, and 83.1% of them were under 7 years of age. Four types of enteroviruses were isolated from 38 of 59 cases (64.4%), and virus type isolated were coxsackie B1 (15 cases), B4 (2 cases), B5 (4 cases), echovirus 16 (14 cases), a mixture of coxsackie-virus B4 and echovirus 16 (2 cases) and an unidentified strain (one cases). Virus types isolated from cerebrospinal fluid were coxsackievirus B1 and B5. In the serological studies of 47 paired sera of patients, a significant rise in titer against 4 virus types isolated was 29.8% with coxsackievirus B 1, 12.8% with coxasackievirus B4, 17.0% with coxsackievirus B5, and 34.0% with echovirus 16. In relation between virus type isolated and a significant rise in titer of neutralizing antibody, almost all of the patients' sera (80.0%) exhibited a significant rise in titer of neutralizing antibody against one isolated virus type, but some patient's sera (20.0%) exhibited a significant rise against two or more of the viruses.
The chemical and biological characteristics of Soluble Protective Antigen (SPA) separated from culture fluids of Salmonella enteritidis strain 2547 were analysed. It was shown that SPA has 3-hydroxy, nonpolar fatty acids by thin-layer chromatography. The fatty acids were identified as lauric, myristic, palmitic and 3-hydroxymyristic acid using gas chromatography-mass spectrometry and mass chromatography. These fatty acids are common constituents of the lipid A obtained from S. enteritidis. SPA was found to enhance the plaque-forming cell (PFC)-response to sheep erythrocytes in the recipients' spleen. In addition, SPA enhanced the clotting activity of Limulus amebocyte lysate. These results show that SPA possesses the properties of lipopolysaccharides isolated from strain 2547 by chemical procedures.
During the period from May to August, 1984, an epidemic of infectious disease with echovirus type 16 occurred in Tono area of southeast in Gifu prefecture. This virus caused children to have different clinical symptoms, one was a exanthem disease and another was aseptic meningitis. These cases comfirmed by virological and serological methods were 48 cases, that is, patients withaseptic meningitis were 24 cases and patients with exanthem disease were 24 cases. By serological examination of antibody against echovirus type 16, it was confirmed that this virus type invaded Gifu Prefecture before 1984.
On one hundred thirty seven cases with aseptic meningitis and 36 cases except aseptic meningitis in 1988, virological and serological investigations were performed. Cases with aseptic meningitis ranged from 0 to 15 years old age, and 90.5% of them were under 8 years of age. Seven types of enteroviruses and one unidentified strain were isolated from 58 of the 137 cases (42.3%), and echovirus type 18 was recoved from 46 cases (79.3%). Echovirus type 18 was isolated also from 16 cases with acute febrile illness and exanthem etc. With the cross neutralizing test between the prototype and isolated strains in 1988 of echovirus type 18, a slight antigenic variation could be found. In the serological studies in 1989, the positive rate of neutralizing antibody to echovirus type 18, was rised remarkably in children under 8 years old.
Five cases of the respiratory infections, in which Neisseria meningitidis was isolated upon the examination of the transtracheal aspiration (TTA), were reported. Patients studied were four males at the ages of 18, 56, 66 and 78 years, and a 18-year old female. Five cases were bronchopneumonia (two cases), acute bronchitis (two cases) and diffuse panbronchiolitis (DPB) (one case). Underlying diseases were as follows: lung cancer or acute myocardial infarction in cases of bronchopneumonia, and interstitial pneumonia or pulmonary tuberculosis in cases of acute bronchitis. There was a case in which only N. meningitidis was clutured from specimens obtained by TTA, although anothor different organis, in addition to N. meningitidis, was recovered from the patient of other cases. Other organisms found together with N. miningitidis were H. influenzae (2 cases), S. dysgalactiae (1 case) and M. tuberculosis (1 case). Predisposing factors were common cold in the female patient and bronchoscopic examination in the cases of lung cancer and of interstitial pneumonia. Two of the five cases occurred consecutively in the same room and so they were considered as nosocomial infections. With these findings, it can be posturated that N. meningitidis might be one of the etiological agents of the respiratory infections.
Chlamydia trachomatis infection in the general population has recently been attracting attention. In this regard, we examined the incidence of positive antigen in the genital organs in pregnant female women in order to investigate the prevalence of this infection. EIA (Chlamydiazyme) was performed in 5, 000 married pregnant women, and in 317 unmarried women who underwent artifical termination of pregnancy. The study was carried out between June 1986 and September 1989 in Sapporo City (3, 932 subjects), Kushiro City (328 subjects), Muroran City (280 subjects), Kitami City (357 subects), Kucchan Town (220 subjects) and Urakawa Town (200 subjects). 1. Among married pregnent women, the detection rate of C. trachomatis was 6.1%(222/3, 666) in Sapporo City, 7.3%(24/328) in Kushiro City, 6.1%(17/280) in Muroran City, 7.8%(24/306) in Kitami City, 6.8%(15/220) in Kucchan Town and 7.5%(15/200) in Urakawa Town, showing no particular difference according to the area. The overall detection rate was 6.3%(317/5, 000). 2. The rates of detection of C. trachomatis antigen by EIA were 22.9%(61/266) and 15.7%(8/15) for the unmarried women who underwent artifical termination of pregnancy in Sapporo and Kitami, respectively. Both rates were higher than those in the married pregnent women. This result suggests the prevalence of C. trachomatis among young women showing a high level of sexual activity. 3. Among married subjects, the detection rate was 21.3% for those in their late teens, 8.9% for those in their early 20s, 6.0% for those in their late 20s, 3.7% for those in their early 30s, and 2.9% for those 35 years or older; thus, the younger they were, the detection rate became elevated. The above findings show that latent epidemics of C. trachomatis infection are present to a considerable extent among young women.
The type-specific polysaccharide antigen of the group B streptococcus (GBS) type Ia as extracted and purified according to the procedures of Kane and Karakawa. Using this purified polysaccharide antigen, we made a sensitive and specific assay system of enzyme-linked immunosolvent assay (ELISA) and measured the titres of of the type-specific antibodies in materanl sera and cord blood sera. The titres of antibodies in 78 pregnent women (26 Ia carriers, 18 other types of GBS carriers and 34 non carriers) were compared. A mother of an infant affected by early onset infection of GBS Ia had a titre of antibody 1: 10 at delivery, while 2 years later she became a non carrier and had a titre of antibody over 1: 160. The titres of antibodies in 27 pair sera of mothers and cords were well correlated.
Sensitive reverse transcriptase assay was applied to human immunodeficiency virus type 2. The kinetics of this assay, stability of the enzyme and the effect of BSA to this assay indicated that the condition of this assay should be 37° in reaction temperature. The sensitivity of this assay increased by adding more than 10μg/ml of BSA. The sensitivity of this assay is at least four times more than that of CPE assay using Molt-4 cell. Other HIV-2 isolate, LAV-2 in culture medium was also detectable in this condition. Moreover reverse tanscriptase inhibiting antibody that specifically inhibits HIV-2 reverse transcriptase was found by this assay.
We evaluated the opsonic activities of human intravenous immunoglobulin (IVIG) preparations against pathogenic organisms by measuring the luminol-enhanced chemiluminescence (CL) from human polymorphonuclear leukocytes (PMN) during the phagocytosis of these organisms. Polyethylenglycol-treated IVIG significantly increased the CL of PMN by opsonization against various bacteria and Candida albicans (p<0.01). The high CL induced by IVIG with intact Fc-fragment showed no significant differences among lots or preparations made by different treatments. In both PMN-CL in the absence of human serum and whole blood CL at low concentration of serum complement, the CL response was not affected by pepsin-treated IVIG. In the severely burned patients' blood with a very low concentration of serum immunoglobulin, IVIG with intact Fc-fragment significantly increased CL (p<0.01). It is suggested that administration of IVIG is useful against infections in these patients. The measurement of whole blood CL in vitro may be useful for evaluating the opsonic activity of IVIG against target pathogens in vivo.
We devised a Citrate-Acetate (CA) medium for rapidly differentiating Shigella. The medium consisted of 3.0 g of sodium citrate, 2.0 g of sodium acetate, 0.2 g of glucose, 1.0 g of dipotassium phosphate, 1.0 g of mono ammonium phosphate, 0.2 g of magnesium sulfate, 5.0 g of sodium chloride, 0.08 g of brom thymol blue, 15.0 g of agar, and 1000 ml of distilled water. An evaluation was made of the CA medium, for the rapid differentiation of 23 Shigella strains, 129 Escherichia coli strains and 130 isolates, that formed colourless colonies suspected to be Shigella on SS agar plate, from feces of healthy people. The results obtained were as follows 1) On the CA medium, all Shigella strains did not grow and there was no change in colour. 2) Positive growth rates of E. coli strains after incubation for 24 hr at 37° on CA medium, sodium acetate medium (Acet) and Christensen citrate medium (C-Cit) were 96.0%, 95.2% and 28.0%, respectively. Therefore, the positive growth rate of E. coli strains after incubation for 24 hr on CA medium was significantly higher (p<0.01) than that on C-Cit medium. 3) Positive growth rates of isolates after incubation for 24 hr at 37°C on CA medium, Acet medium and C-Cit medium were 95.4%, 83.1% and 71.5%, respectively. Therefore, the positive growth rates of isolates after incubation for 24 hr on CA medium was significantly higher (p<0.01) than that on Acet medium and C-Cit medium. 4) The genus of tested isolates were Escherichia, Enterobacter, Hafnia, Pseudomonas, Citrobacter, Aermonas, Alcaligenes and Morganella. 5) It was found that sodium acetate, and sodium citrate and glucose incorporated into the CA medium had enhanced the growth of Escherichia and Hafnia, respectively. From the above results, the CA medium is recommend as a differential medium for use in the rapid differentiation of Shigella and organisms that formed colourless colonies suspected to be Shigella on SS agar plate in the routine examinations.
Pasteurella multocide (P. multocida), a small gram-negative bacillus, has been known to be the causative agent of hemorrhagic septicemia in animals. P. multocida infection in human was reported as skin abscess and/or septicemia after an animal bit or scratch. Pulmonary infections of P. multocida have been developed in the patients with chronic pulmonary diseases such as bronchiectasis. In Japan, however, P. multocida respiratory tract infections are rare. In this report, a 80-year-old female with bronchiectasis was admitted on August, 1985. She had a productive cough, hemosputum, and a low grade fever. The chest X-P on admission showned an atelectasis of the left middle lobe and severe bronchiectatic changes of the left lower lobe. P. multocida was isolated from her sputa. The chemotherapy of CTM resulted in clinical imporvement. On May 1988, she complained of a productive cough and a low grade fever again. P. multocida was isolated from the sputum on several occasions in significant numbers (1×108/ml). Recently, the cases of the chronic respiratory diseases have been increasing. We think, P. multocida is important and should be considered as a pathogen in the care of chronic pulmonary diseases.