We evaluated the incidence of antibodies against canine distemper (CD) virus, canine parvovirus, Toxoplasrna gondii (T gondii), Aujeszky's disease virus, porcine reproductive and respiratory syndrome virus, Rickettsia tsutsugarnushi, Rickettsia japonica, and Coxiella burnetii in 30 raccoon dogs and 5 deer captured in suburban areas in Kanagawa Prefecture. Among theraccoon dogs, anti-T. gondii antibody was detected in 1 animal (3.3%) and anti-CD virus antibody in 9 (30.0%). However, antibodies against the other pathogenic microorganisms were not detected in the raccoon dogs. The deer had none of the examined antibodies.

View full abstract

We isolated strains of Helicobacter pylori from gastric mucosa of patients with peptic ulcer before and after eradication therapy, and studied their sensitivity to amoxicillin (AMPC) and clarythromycin (CAM).
Of 85 strains of H. Pylori isolated before therapy, MIC₉₀ was 0.025μg/ml and no strains were resistant to AMPC. On the other hand, MIC₉₀ of CAM was 0.05μg/ml and seven (8.2%) were already resistant to CAM.
The H. Pylori strains from eight cases of failed eradication therapy with lansoprazole+ AMPC remained AMPC sensitive. However H. pylori strains from nineteen cases (82.6%) out of 23 of failed eradication therapy with lansoprazole + CAM became CAM resistant.
The situation was similar for the cases of failed eradication therapy with lansoprazole +AMPC+CAM.
Drug sensitivity tests prior to eradication therapy are to be recommended. A disc method may be used as a simple alternative to MIC measurement.

View full abstract

To evaluate the contribution of the subsets of T helper lymphocyte (Th) to the development of pulmonary lesion in mycoplasma pneumonia, we compared the pathological findings between Thl dominant mice (C57BL/6) and Th2 dominant mice (BALB/c) in experimental Mycoplasma pulmonis (M. pulmonis) pneumonia. Mice (ICR, C57BL/6, and BALB/c) were intranasally inoculated with 0.03 ml of a solution containing M. pulmonis (1×10⁸) colony forming units per ml. Another M. pulmonis inoculated ICR mice were treated with interleukin-2 (IL2; 4.8μEg/day), days 3-9, intracutaneously. All mice were sacrificed at day 14, and the lung specimens were examined. Peribronchial lymphocyte cuffing was more prominent in C57BL/6 mice than that of ICR mice, and intra-alveolar inflammatory-cell infiltration in BALB/c mice was more prominent than in ICR mice. Pathological patterns of the lung in IL-2 treated ICR mice were mimicking those of C57BL/6 mice. These results suggested that pathological patterns of mycoplasma pneumonia in mice might be altered by the imbalance of host T helper lymphocyte subset.

View full abstract

In this study we assessed the usefulness of Caco-2 cells, derived from a human colon carcinoma, to isolate an influenza virus.
Throat washings collected from 30 patients with influenza-like illnesses in Miyazaki Prefecture in 1997 were inoculated in MDCK and Caco-2 cells, 17 influenza virus strains were isolated in MDCK cells, and 20 in Caco-2 cells. Of all the viruses isolated, only one strain was identified as influenza virus type B; other strains were identified as type A (H3N2). Furthermore, some influenza viruses were isolated in Caco-2 cells also from the specimens collected between 1991 and 1997. With Caco-2 cells, each type of influenza virus was isolated effectively without the supplement of trypsin in the culture medium.
These facts indicate the usefulness of Caco-2 cells as a host to isolate influenza virus as shown to be suitable in the detection of many types of enteric viruses. Caco-2 cells will serve as a useful cell line for the surveillance of infectious disease because Caco-2 cells are sensitive to a wide range of virus.

View full abstract

During the 7 years from 1990, thirty-two patients (20 in male and 12 in female, mean age; 53 years old) were diagnosed as having pulmonary cryptococcosis. To clarify the essential points for early diagnosis of pulmonary cryptococcosis, we reviewed the clinical records and chest images. Three patients had a past history of pulmonary tuberculosis and eleven patients had underlying disorders such as malignancy, chronic pulmonary diseases and so on, but no HIV infection, which would affect this disease. Eighteen patients did not have any past history nor complications.
The symptoms such as cough, sputum, chest pain and fever were generally of low-grade, 14 patients had no symptom at diagnosis. Except of some patients with severe infections and severe underlying disorders, laboratory findings such as inflamatory and nutritious markers were almost within near the normal range. On plain chest X-ray films the distribution of lesions was almost in proprtion to the volume of the lobes. The multifocal nodular and/or infitrative shadows wer observed in about 2/3 cases and single lesion in about 1/3. The width of lesions were minimal except of one case with interstitial pneumonia and two cases with multifocal segmental pneumonia. The cavity lesions were observed in 7 cases and hilar lymphadenopathy in 3 cases.
On CT images, the lesions were almost located in the outer zone, the lesions which wereadjacent to the pleura were observed in 15 cases. Cavitary lesions were almost smooth in edge and ubiquitous, the walls were also thick. The peripheral air-bronchogram in the nodular/infitrative shadows were observed in three cases.
Pulmonary cryptococcosis is air-borne and almost a chronic infection except in AIDS patients, so careful planning for examination is essential with considerations of the characteristics of clinical and imaging features of this infection.

View full abstract

The antibody responses of 31 elderly persons (11 males and 20 females, mean 82.2 years) and 23 adults (mean 40.7 years) to inactivated influenza vaccine were studied. Participants were given twice the influenza vaccine containing A/Yamagata/32/89, A/Kitakyusyu/159/93, and B/ Mie/1/93. Serum specimens were obtained before vaccination, and 2 weeks and 3 months after the second vaccination. Antibody to influenza was determined by hemagglutination inhibition test (HI).
The rates of fourfold or greater increases of HI antibody agaisnt the A/Yamagata, A/ Kitakyusyu, and B/Mie were 96.8%, 100%, and 74.2%, and there was no significant difference of the rates of fourfold or greater increases between elderly persons and adults. The rates of possession of HI antibody more than 128 in titer after vaccination were significantly higher than those of HI antibody before vaccination, and were maintained at high levels up to three months after the second vaccination.
These results suggest that antibody response to the inactivated influenza vaccine is satisfactory in the elderly.

View full abstract

A Shigella flexneri 2a strain, which did not respond clinically or in laboratory tests to treatment with new quinolone derivatives, was isolated for the first time in Japan from a patient admitted for diarrhea to a Tokyo hospital. The mechanism of resistance was examined by sequencing the quinolone resistance determining region (QRDR) of the gyrA (a quinolone target enzyme) gene and by comparing the active efflux mechanisms of two strains isolated from this patient (before hospitalization and after tosufloxacin treatment) and one strain isolated from a patient with secondary infection with that of the standard strain ATCC 29903. DNA sequencing revealed that two amino acid substitutions, namely, Ser (TCG)-83 → Leu (TTG) and Asp (GAC)-87 → Gly (GGC), had occurred in the gyrA of all 3 strains isolated from the patients. Examination of the accumulation of antibiotics in these 3 strains revealed that the strain isolated after tosufloxacin treatment had the highest resistance to tosufloxacin, and exhibited decreased accumulation of tosufloxacin in the bacterial cells discharged after 5 days of treatment with this antibiotic. However, accumulation was restored by addition of a proton-pump inhibitor. These results suggest that the strains isolated from the inpatient and the patient with secondary infection acquired resistance due to dual gyrA mutation induced by treatment with new quinolone antibiotics. Furthermore, in addition to this dual mutation, the active efflux mechanism also appears to be associated with resistance in bacteria that have been exposed to tosufloxacin.

View full abstract

We examined the therapeutic effect of cefluprenam (CFLP) on the polymicrobial urinary tract infection associated with infectious stones as compared to that of ceftazidime (CAZ), using the experimental polymicrobial urinary tract infection caused by Proteus mirabilis (P. mirabilis), Pseudomonas aeruginosa (P. aeruginosa) and Enterococcus faecalis (E. faecalis). In order to form bladder stones in rats, a sterile zinc ring was implanted as a foreign body, followed by inoculating P. mirabilis E05106 transurethrelly. Thereafter, P. aeruginosa E030033 and E. faecalis 966 were inoculated according to the same way. CFLP or CAZ was administered intravenously twice a day for 5 days. The therapeutic effect was evaluated by the eradication of these bacteria from the urine, the kidney and stones, inhibition of growth of stones, and also a value of BUN. The urinary excretion rates of CFLP and CAZ was also determined (20 mg/kg). In result, CFLP significantly more effective in the eradication of P. mirabilits, P. aeruginosa and E. faecalis from the urine, the kidney and stones than the control than the control (p < 0.05). Furthermore, CFLP significantly more effective in the eradication of P. aeruginosa from the urine and also E. facalis from the urine and stones (p < 0.05). Meanwhile, CAZ significantly more effective in the eradication of P.mirabilis from the urine, the kidney and stones and also _{P. aeruginosa} from the kidney and stones than the control. However, CAZ did not show eradicative effect on E. faecalis. The urinary excretion rates of CFLP and CAZ at rats were 59.3% and 59.4%, respectively, within 8 hrs after administration, showing a similar excretion pattern. CFLP exhibited the prominent therapeutic effect on polymicrobial urinary tract infection associated with infectious stones caused by P. mirabilis, P. aeruginosa and E. faecalls. On the basis of these results, it has been strongly suggested that CFLP is highly beneficial in the management of intractable polymicrobial urinary tract infectious in clinic.

View full abstract

This study was investigated to eludidate the clinical role of immune complexs (IC) formed by alginate of persistently colonized mucoid strain in chronic aiway infection.
1. Clinical Observation
Twenty-four cases colonized with mucoid strain and 16 cases with non-mucoid strain were enrolled. Their serum indexes to anti-alginate specific IgG subclass antibodies and level of IC were determined by enzyme linked immunosorbent assays. Significantly higher level of the IgGl, IgG3 and higher level of IC were observed in the group of mucoid positive cases. Deposition of IgGl, IgG3 and IC on the affected part of interstitial lung tissue in a case of persistent infection with mucoid Pseudomonas aeruginosawas detected.
2. Experimental Observation
Ninety mice immunized with alginate were used. They were intubated with 20μg/body of alginate, 4-106 colony forming unit/body of mucoid Pseudomonas aeruginosa PT1252, or 40μl/ body of IC into each group. Then after Fcγ receptor (FcγR) on neutrophils in bronchoalveolar lavage fluid was analyzed by flow cytometric technique. Expression of FcγR on neutrophils was inhibited against alginate intubation and IC one. In contrast, FcγR expression was increased at initial phase after mucoidPseudomonas aeruginosaintubation, which was influenced by nonspecific activation of neutrophils due to bacterial infection.
3. Conclusive Words
From the above, The expression of FcγR on neutrophils was inhibited by alginate. It may be that IC deposition on lung tissue in chronic airway infection with mucoid strain was persisted by poor binding ability of neutrophils to IC, in spite of a large numbers of neutrophils accumulation. Consequently, the immune-reaction induced by alginate makes clinical prognosis of such patients more intractable.

View full abstract

This study was performed to demonstrate the pathogenetic role of mucoid-alginate produced by Pseudomonas aeruginosa in relation to immuno-interaction in host cells
The obtained resutls are as follows.
1. Clinical Observation
The concentration of MPO, TNFα, and IFNγ in peripheral blood of 37 cases of chronic airway infection were observed. The cases were divided into the groups with mucoid strain positive (21 cases) and negative (16 cases), and each group was also divided into subgroups with active clinical symptom and without.
High concentration of serum IFNγ was shown in the group of mucoid strain positive cases (0.10±0.08 IU/ml in healthy volunteers group, 0.54±0.56 IU/ml in mucoid group with symptom: p <0.01, 0.09±0.15 IU/ml in mucoid group without symptom, 0.13±0.13 IU/ml in non-mucoid group with symptom, 0.09±0.13 IU/ml in non-mucoid group without symptom). But plasma concentrations of MPO and serum concentrations of TNFα were not increased in all groups, except MPO in non-mucoid group with symptom. It indicates that IFNγ has a haracteristic role in chronic airway infection with mucoid Pseudornonas aeruginosa (P. aeruginosa).
2. Experimental Observation
One hundred twenty-two of alginate-immunized mice were made by 5 times of intraabdominal injection with purified alginate extracted from mucoid P. aeruginosa PT-1252. 4± 10₆ CFU/body of mucoid P. aeruginosa PT-1252 or 20μg/body of purified alginate was intubated per trachea into the immunized mice, and changes in mean fluorescence intensity (MFI) of MPO and TNFα on neutrophils and IFNγ on CD3 positive lymphocytes in BALF were observed
MFI of MPO (99.7±16.5→4177.0±23.1: p<0.001) and TNFα(59.2±13.3→197.0±62.0: p<0.001) were increased on day 2 and they were kept up till on day 30 in P. aeruginosa group. But in alginate group they were temporally decreased on day 2, and they were gradually increased (MPO: 99.7±16.5→212.6±12.1: p<0.001, TNFα59.2±13.3→162.4±30.9: p< 0.01). The temporally decrease of MPO and TNFγ on day 2 in alginate group may suggest that a large amount of alginate inhibits neutrophils chemotaxis. MFI of IFNγ in P. aeruginosa was also increased (110.0±32.9→198.3±23.0: p<0.01) on day 2 and it was gradually decreased on day 30 (156.0±13.8). In alginate group, MFI of IFNγ in BALF was increased on day 5 (110.0±32.9→201. 0±49.3: p <0.001) and kept a high level till day 30 (223.0±57.5: p <0.01). In morphological, the moderate grade of lymphocytes infiltration observed in intersitial area of lung tissue on both groups of P. aeruginosa and alginate intubation mice. The findings was continuously seen from day 5 through day 30. The lymphocytes infiltration appeared in lung tissue can be though as an result of immuno-interaction with IFNγ positive lymphocytes and mucoid-alginate.
Inferences are drawn from the clinical results and the experimental ones that the CD3 positive IFNγ Alymphocytes expressed by immunological interaction with mucoid-alginate and the IFNyγ Alymphocytes (Thl like cells) plays an important role on advancement of chronic airway infection with mucoid P. aeruginosa.

View full abstract

A 56-year-old male without respiratory symptoms, past history nor familial history, was admitted to our hospital because of pulmonary solitary nodule on the chest radiograph. Computed tomograph showed a smooth surface nodule in the left lower lobe (segment 8). Bronchofiberscopy could not give any specific histological findings nor bacteriological findings, therefore the patient underwent partial pulmonary resection by videoassisted thoracoscopic surgery. Pathology of the resected specimen revealed epithelioid cell granuloma with giant cells but not any acid-fast bacilli. He was treated with rifampicin and isoniazid for “tuberculoma”. After three weeks, thirty colonies grew on an Ogawa's egg medium, which were identified as Mycobacterium avium by PCR.
There are few reports about the solitary pulmonary nodule due to M. avium-intracellurale complex infection, however, it is supposed that there are many “tuberculoma's” without bacteriological differentiation between tuberculosis and nontuberculous mycobacterial infection.
It is difficult to diagnose a solitary pulmonary nodule caused by nontuberculous mycobacterial infection with bronchofiberscopy, and the location of the lesion is usually subpleural, therefore it is thought that videoassisted thoracoscopic surgery is valied for the diagnosis of solitary pulmonary nodule due to nontuberculous mycobacterium.

View full abstract

We report here a 2-year-old boy with a Streptococcus intermedius brain abscess and bilateral ventriculitis successfully treated with a high dose penicillin G (200, 000 U/kg/dose, 6 times a day, 1 hour continuous infusion). Although hydrocephalus residued, the high dose penicillin G therapy cured his brain abscess and bilateral ventriculitis. The minimal inhibitory concentration of penicillin G to the isolate was 0.008μg/ml. The penicillin G concentration in the cerebrospinal fluid after 2 hours from the infusion was about 5μg/ml. S. intermedius must be considered as one of the causative agents for brain abscess. High dose penicillin G therapy is one choice of treatment for brain abscess due to penicillin-susceptible streptococci.

View full abstract

[in Japanese]

View full abstract