We studied the differences among clinical isolates of Penicilin-resistant Streptococcus pneumoniaeby polymerase chain reaction (PCR) of pbp2bgene, followed by serotyping and pulsed field gel electrophoresis (PFGE) analysis. Clinical isolates were recovered from sputum samples from patients with respiratory infection in Tottori University Hospital between June 1986 and May 1996. By PCR, altered pbp2bgenes of the resistant isolates were detected in 76.5%.The percentages of the isolates that had altered pbp2b genes increased concomitantly with the minimum inhibitory concentration (MIC). By serotyping the percentage of 19F, 23F, 6B and 14 was 54.5%, 18.2%, 9.1% and 9.1% respectively. The frequency of isolates resistant to penicillin increrased rapidly from 1991 in this hospital and most isolates belonged to serotype 19F. The resistant isolates in this hospital and 4 clinical resistant isolates of S. pneumoniae 19F in a second hospital were studied by PFGE. 14 of 18 resistant 19F isolates in this hospital and all 19F isolates from the second hospital presented an identical pattern. The remaining 4 samples were similar though not complietely identical. These results indicate that the penicillin resistant 19F isolates have a common clonal origin and have spread rapidly from 1991 in this hospital.

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In the winter of 1996, a total of 71 working adults from 20 to 60 (m=37.3) years of age were immunized with one or two doses of inactivated influenza vaccine formulated in Japan. The reciprocal geometric mean hemagglutination inhibition antibody titers [mean HI] and the percentage with a hemagglutination inhibition titer of≥×128 [percent ≥×128], against A/Yamagata/32/89 (H1N1), A/Uhan/359/95 (H3N2), and B/Mie/1/93 were significantly elevated by either doses. The pre-/post-vaccination mean HI of H1N1 were 61.5/668.4 with one dose, and 85.7/637.3 with two doses. The percent≥×128 were 36.5/96.2 with one dose, and 47.4/94.7 with two doses. No significant differences were apparent between one and two doses. Similarly, nonsignificant differences were observed for the other two antigens contained in the vaccine. We observed the sufficient immune response to only one dose of inactivated influenza vaccine, which emphasizes the merit of one dose method of influenza immunization

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Adhesion of Helicobacter pylori to both human gastric carcinoma cell lines (MKN45, MKN28 and KATO III) and prepared primary human gastric epithelial cells were analyzed with flowcytometry. All strains adhered to human gastric carcinoma cells. Especially, these strains strongly adhered to MKN45 cells. Adhesion of H. pylon strains to prepared primary human gastric epithelial cells was also observed. However, the adherence rates of H. pylonto these cells were different among the cells used. These results suggested that the host factor might be important for adhesion of the bacteria to human gasgric cells. In addition, H20 monoclonal antibody directed to H. pyloriHSP60 inhibited the adhesion of H.pylonto both cells. These results indicate that H. pyloriHSP60 might be associated with the adhesion of the bacteria to human gastric epithelial cells.

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To determine the cause of nosocomial infections, all febrile episodes of hospitalized elderly patients aged 65 and older at a hospital in Fukuoka City were categorized between April 15 1994 and April 14 1995.
A febrile episode was a temperature above 37.5°C after 7 consecutive days of normal body temperature (below 37.5°C). Various clinical tests including blood examination, urinalysis, chest radiography and bacterial culture were done on the first and 7th day of the fever.
A total of 1105 episodes in 443 patients (male 136, female 307) fulfilled this criteria for fever. The fevers were mainly due to respiratory tract (381 cases, 34.5%) and urinary tract infections (263 cases, 23.8%). There were 135 infections (12.2%) of other kinds and 297 cases (26.9%) that were classified as unknown.
Approximately 70% of the febrile episodes were caused nosocomial infections, suggesting increased risk of infection in the hospitalized elderly and the importance of early detection of febrile changes in elderly inpatients.

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Five Shigella strains isolated from stool cultures of imported diarrheal cases in Japan, did not react to any antisera of the established Shigella serovars.
These strains had the typical biochemical characteristics of Shigella dysenteriae, and were biochemically identical. All strains were positive in the Sereny test and other tests for invasivness; these indicate that they can cause shigellosis in humans.
The results of antigenic analysis revealed that they did not belong to any of the recognized or provisional serovars, and were serologically indistinguishable.
They had the same drug-resistance pattern (CP·TC·SM·ABPC·ST) and plasmid-profile.
Strain 96-204 is designated as the test strain for this new serovar.

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The Roche Amplicor Mycobacterium Tuberculosis Kit ^® (Amplicor PCR) and Gen-Probe Amplified Mycobacterium Tuberculosis Direct Test ^® (MTD) were compared for efficiency in the direct detection of tubercle bacilli in sputum specimens. In total, 72 sputum specimens obtained from patients with active tuberculosis were examined with the Amplicor PCR and MTD kits. Of the 72 test specimens, 45 and 41 samples were positive by the Amplicor PCR and MTD methods, respectively. These values were larger than those obtained by the culture method (Septi-Chek^®) and acid-fast bacilli-smear test, yielding 37 and 20 positive results, respectively, and the sensitivities were as follows: smear, 39.2%; culture, 72.5%; Amplicor PCR, 88.2%; MTD, 80.4%. Amplicor PCR was similarly or somewhat more sensitive in direct detection of M. tuberculosis in sputum samples from patients with clinically active tuberculosis than was MTD.

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Lyme borreliosis is one of the zoonotic and vectorial diseases in the world. Female of Taiga tick, Ixodes persulcatus play an important role being the main vector of the agents, Borrelia burgdorferi sensu lato, of the disease in the Far East including Japan.
The number of patients who contacted Lyme borreliosis have been estimated to be around 100 cases in all of Japan. We also detected the spirochetes from the skin tissues of 27 cases involving erythema and 3 cases without erythema after tick bites in Hokkaido. We noticed that in those cases when the victims removed ticks by themselves using their fingers within several days later, almost all of them developed erythema.
On the other hand, an infected tick from 8 cases which were removed by surgical procedure, the spirochetes were not transmitted in these cases (No.31-38). Therefore, we supposed that the transmission of agents takes place by regurgitation of the spirochestes from the midgut diverticula due to the pressure by fingers (calling Syringe phenomenon). Then, the authors recommended that fine tweezers has a firm grip around the mouth part of infested tick and pull out within 24 hours and/or trials to surgical operation for tick removal on passing over 24 hours after infestation.

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Pulmonary diseases are important cause of morbidity and mortality in patients with PM/DM. Thirteen (27%) out of 49 PM/DM patients in the study had developed respiratory failure. Respiratory failure resulted form interstitial pneumonitis (IP) in 6, pulmonary infection in 2 and both in 5 patients with PM/DM.
Respiratory failure was fatal in PM/DM patients with pulmonary diseases and eleven of the 13 patients expired. More importantly, 2 PM/DM patients with respiratory failure had responded to chemotherapy, if it was due to pulmonary infection. Accordingly, it is atmost important to distinguish IP and infection for the cause of respiratory failure. However, plain chest X-ray as well as standard laboratory tests failed to differentiate IP and pulmonary infection. On the other hand, high resolution CT of the lungs, serum endotoxin and serum β-D-glucan were found to be useful for the differentiation of these conditions associated with respiratory failure in PM/DM patients. And additionally low serum level of IgG and lymphopenia at the onset of respiratory failure may suggest that the patients may have pulmonary infection rather than IP.

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Polymorphonuclear leukocytes (PMN) play important roles in the prevention of infection at an early stage. We studied the function of these leukocytes using rabbit models of mandibular bone infection to evaluate the conditions which could not be reproduced in human beings Streptococcus milleri NCTC7331 and Bacteroides fragillis NCTC9343 were inoculated into the mandibular bone of rabbits using the Satoh-Heimdahl method, to produce supposed multiple infection models. Rabbits inoculated with these bacteria were used as a test group, and animals with surgically induced inflammation were used as a control group. We compared the number of leukocytes, the function of PMN, and histopathologic findings.
1) The number of leukocytes increased after treatment, reached a perk on day 3, gradually diminished later, but remained slightly higher than the baseline level on day 7, with persistence of inflammation in both groups.
2) Adhesiveness, ability to migrate and NBT reduction, were accelerated in both groups.
3) These functions of PMN accelerated more in the test group because the bacteria inoculated induced stronger inflammatory reactions and activated a series of infection defense mechanisms in the hosts.
4) Histopathologic examination after treatment showed invasion of inflammatory cells, predominantly leukocytes, in both groups, but heavier and more extensive innitration in the group treated with the bacteria.
All measurements were higher in the test group than the control group. These results showed that in the test group, causative or accompanied microorganisms activated the host's infection defense mechanisms and accelerated the functioning of PMN at an acute stage of infection.

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Biological activities of two strains of Staphylococcus aureus (S. aureus), KU-1-06-37 and KU-1-12-44, which produce enterotoxin type C, TSST-1 and α-toxin were examined using Std: ddY strain mice. These two strains were found to be different in lethality, ratio of weight loss, induction of leukopenia, adhesion to surrounding organs and clearance period of bacterial cells from the liver, kidney and spleen within 24 hrs after intraperitoneal injection in the mice. All of them were weak or fast in KU-1-12-44 injected mice. Serum amyloid A on all the KU-1-06-37 and KU-1-12-44 injected mice rose within 5 hr to 18 hr. However, this concentration of KU-1-12-44 injected mice was about 40% lower compared with that of KU-1-06-37 injected mice at 21 hr. On the other hand, ability of bacterial adhesion to established cell lines, Vero and HeLa cells, was tested in vitro. Percentage adhesion of KU-1-06-37 was high to both cells, but that of KU-1-12-44 was high to Vero cells and was low to HeLa cells. Adhesion of KU-1-06-37 to HeLa cells that were treated with lipoteichoic acid was about 40% inhibition compared with untreated cells, although that of KU-1-12-44 to them was inhibited only 9%. As the results, identical toxinproduced KU-1-06-37 and KU-1-12-44 showed different biological activities in vivo and in vitro. Not only toxin production but also adhesion to cells or organs in mice may contribute to S. aureus virulence to the host.

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We report two cases of aspergillosis with skin lesions. The first involved a 63-year-old female with acute lymphocytic leukemia. In 1992, she received chemotherapy for acute leukemia. Subsequently, she noted swelling of her leg, a skin biopsy was performed and Aspergillus flavus was isolated. The second patient was a 71-year-old male with myelodysplastic syndrome. In 1996, he developed a massive inguinal abscess from which Aspergillus fumigatus was isolated. The prognosis of aspergillosis was poorer in the case with skin swelling than in the one with a massive abscess.

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A 81-year-old man who complained of fever and disturbance of consciousness was admitted to our hospital. Listeria monocytogenes type 1/ 2aA was cultured from only the blood. He was treated with γ-globrine and sensitive antibiotics (PAPM/BP, EM) immediately after admission, and recovered in spite of multiple organ failure due to septic shock. He was not an immunocompromised host and did not have complication of meningitis, but had rhabdomyolysis and liver dysfunction.

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A case of Kartagener's syndrome associated with multiple pulmonary complication was presented. A 19-year-old man was admitted to our hospital because of pulmonary tuberculosis in May 1972. He had been diagnosed as a Kartagener's syndrome because of the presence of chronic parasinusitis, bronchiectasis and complete situs inversus. His chest radiographs in Dec 1972 revealed left pneumothorax. Chest radiographs in Aug 1975 appeared aspergilloma in the right middle lung field. He was administrated intravenous and oral anti-fungal agent and transbronchial instilation of Amphotericin-B because of hemoptysis. Chest radiographs in Jul 1980 resolved the aspergilloma and his symptom were also resolved. In 1996, he had no pulmonary symptoms and respiratory failure.
We consider that the Kartagener's syndrome was good prognosis with adequate pulmonary therapy.

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A 43-year-old female with spherocytosis was admitted to the hospital because of fever, headache, and rapidly progressive anemia. On admittion, she had severe anemia (Hb 3, 9 g/dl) with marked decrease in reticulocytes (0.3%). Some premature leukocytes, such as promyerocyte, were observed in the peripheral blood. Bone marrow smear revealed normocellular marrow with normal presence of erythroid cells. Anti-human parvovirus B19 IgM and IgG antibody was positive in the serum on admission. The DNA of the virus was also detected in the peripheral blood by PCR assay. Thus, this case was diagnosed as acute aplastic crisis caused by human parvovirus B19 infection. Blood transfusion was performed on day 1 and day 2. The symptoms gradually disappeared and increase in reticulocyte (11.5%) was observed on day 5. Premature leukocytes in the peripheral blood disappeared on day 2. She recovered from anemia and was discharged on day 7.

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An asymptomatic 52-year-old male, who was diagnosed as poorly controlled diabetes mellitus (DM), was referred to our hospital because of abnormal shadows on chest X-rays.Physical examinations and laboratory data showed no abnormal findings except increased values of blood sugar and HbAlc (12.1%). Chest X-ray and CT scan revealed multiple nodular shadows in both lung fields. The specimens from transbronchial biopsy showed epithelioid granuloma, and Mycobacterium tuberculosis was confirmed with bronchial wash fluid. The patient was then treated with antimycobacterial drugs and sulfonylurea along with diet therapy. The multiple nodular shadows gradually disappeared. It seemed that impaired host defense mechanisms caused by poorly controlled DM is responsible for the bronchogenic dissemination of M. tuberculosis, resulting in the extension into the mutiple nodular shadows. Thus, a good control of DM is important for the prevention of an onset and excerbation of tuberculosis in patients with DM.

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