The effects of (-) epigallocatechin gallate (EGCg) on the morphology of hyphae and conidia of Trichophyton mentagrophytes were examined using scanning and transmission electron microscopy. EGCg inhibited the germination of conidia and subsequent hyphal growth. After being treated with EGCg for 3 days, conidia showed morphological changes such as deformation, swelling, wrinkles, exfoliation of the surface layer and accumulation of granular, amorphous materials on the conidia and short hyphae surfaces. These morphological changes became apparent day by day. After 5 days, most of the ungerminating conidia were broken down.
The hyphal cell walls were also exfoliated and the cytoplasm in most of the cells disappeared completely. Inner organellas were variable in size and shape. Many high electron dense particles, greatly enlarged vacuoles appeared in the cytoplasm. The degradation of cells occurred.These observations indicates that EGCg causes lysis of the conidia and hyphae.
These findings suggest that EGCg may be used as an antidermatophytic agent against T. mentagrophytesinfection.

View full abstract

An experimental model of Chlamydia trachomatis (C. trachomatis) intrauterine infections was established in mice. Using this model, studies were conducted on the immune response at the site of infection by applying the ABC staining method and monoclonal antibodies directed against each of the various species of immunocompetent cells.
At the same time, the serum levels of C. trachomatis-specific IgA and IgG antibodies were also measured. The immunological correspondence of these serum antibodies to C. trachomatis elementary bodies was also investigated.
1. With regard to the immunological response at the site of infection, there was greater infiltration by T cells than by B cells. Determination of the subsets of the T cells revealed that CD8+ Tcells outnumbered CD4+ T cells. In addition, among the B cell lineage, there was moderate infiltration by IgA-positive B cells, whereas the infiltration by IgM-positive B cells and IgG-positive B cells was very slight.
2. C. trachomatis-specific IgA and IgG antibodies came to be detected beginning on the 7th day of infection. When the immunological correspondence of these serum antibodies to C. trachomatis elementary bodies was investigated by western blot analysis, one band reacted with the major outer membrane protein (MOMP; 39.5 Kd), while a second band reacted with a 60-Kd protein.
On the basis of these findings, it was surmised that, as the immunological response to C.trachomatis infections, the host recognizes the MOMP and a 60-Kd protein in C. trachomatis elementary bodies, thus initiating a series of immune responses. It was also surmised that, at the sites of infection, cells belonging to the T-cell lineage, especially CD8+ cells, play an important role in the host's defense mechanism against the infection by C. trachomatis.

View full abstract

The anti-endotoxin activity of mouse monoclonalIgM antibody E5 was determined against various lipopolysaccharides (LPSs) derived from Escherichia coli J5, E. coli O111: B4, Klebsiella pneumoniae and Pseudomonas aeruginosa by measuring limulus activity by endospecy method. In general, above 0.2μg/ml of E5 reduced almost dose-dependently the limulus activities of all LPSs tested, although the degree of reduction in limulus activity somewhat varied among bacterial species. The limulus activities of LPS from E. coli J5, E. coliO111: B4, K. pneumoniae and P. aeruginosa decreased by 72%, 50%, 44%, and 58%, respectively, by exposure to 5.0μg/ml of E5 for 2 hrs.

View full abstract

This paper evaluates the immune responses of geriatric patients to three vaccines; Japanese Encephalitis vaccine, inactivated Infuluenza vacicne and cold-adapted live Influenza vaccine. To evaluate the immune response, serum antibodies and lymphocyte phenotypes were measured of all three vaccines. In addition, nasal specific IgA antibodies were measured of the cold-adapted live Infuluenza vaccine.
In the case of Japanese Encephalitis vaccine and the inactivated Influenza vaccine patients were inoculated subcutaneously with a 0.5 ml dose of the vaccine respectively. While with the cold-adaptedvaccine, the doses were administered via spray into the bilateral nasal canals in 0.25 ml dosesrespectively.
Measurements were taken both prior to and four weeks after the inoculations. In all the cases, sera were measured utilizing the hemagglutination inhibition (HI) method. Peripheral lymphocyte phenotypes were also measured utilizing the flow cytometry. In the case of the cold-adapted live influenza vaccine, nasal specific IgA antibodies were measured by the ELISA method.
The results were as follows in all three cases:
1. Recipients who showed elevated HI antibody titers of more than 4 folds were 27.7% to 85.0%.
2. Peripheral lymphocyte phenotypes in geriatric patients were the same as the young control group both before and after inoculation.
3. Antibodies to the inactivated Influenza vaccine were maintained at high levels up to three months after inoculation.
4. The significant increased specific IgA antibodies were 50%. In conclusion, this suggests that inoculation of geriatric patients with the above vaccines is both safe and effective.

View full abstract

This is a retrospective study comparing the methods of preventing fungal infection in a total of 420episodes of chemotherapy-induced neutropenia of more than 10 days' duration between 1986 and 1992 in104 patients with acute leukemia and 62 patients who underwent bone marrow transplantation.
The episodes were divided into five groups according to the prophylactic regimens group 1, oral amphotericin B (OA) only; 2, OA and nebulization of amphotericin B (NA); 3, OA and NA with a laminar air flow system (LAF); 4, oral fluconazole (OF) and NA; 5, OF, NA, and LAF. The total numbers of neutropenic episodes were 115, 141, 95, 37, and 32, respectively.
A total of 15 episode of invasive fungal infections were documented. Aspergillosis was seen on two occasions each in groups 1 and 2, while none was seen in the patients who were under the LAF system.
Nebulization of amphotericin B did not have a significant preventive effect in this study and it needs to be evaluated further by a randomized study.

View full abstract

We have attempted to examine the cause of lowering phagocytic activity of phagocytes on clearance of bacteria in SLE patients. The experiments were performed in SLE model mice, NZB/WF1 female mice (BWF1). The ratio of the cells which phagocytized fluorescent microspheres and possessed Mac-1 on both peripheral blood leukocytes (PBL) and peritoneal macrophages (PMo) in BWF1 female mice was shown decreasing by using the flow cytometric method before the present of typical symptoms of SLE (under 25 weeks-old), i.e., the appearance of ANA and IgG deposits in glomeruli. In a period of showing the symptoms of SLE (over 25 weeks-old), the ratio of Mac-1 positive cells on PBL increased, but phagocytic function of these cells did not increase. On the other hand, on PMo, they decreased together in that period. Moreover, the phagocytosis of PMo in 15, 25, 35 weeks-old mice against fluorescent microspheres showed a significant increase (1.7 times) by replacing opsonin with normal (ddY) murine serum instead of BWF1 murine serum, and the phagocytosis of PMo treated with anti-Mac-1 antibody (M1/70) showed a significant inhibition (46%) against that of untreated PMo.
These results indicate that lowering phagocytic activity of phagocytes on primary defence against microbial infections in SLE patients is due possibly to decrease of C3 activity in the serum rather than a decrease of positive percentage and function of Mac-1 on these cells.

View full abstract

We examined biological properties of strains of methicillin-resistant Staphylococucs aureus (MRSA) which were isolated in our ward in 1991 and 1992. A total of 47 MRSA strains were isolated in 1991 and 64 in 1992. The majority of these strains of MRSA were highly resistant to DMPPC, CEZ and IPM, and were intermediately resistant to MINO. All these strains were, however, sensitive to VCM. The number of coagulase type II strains increased from 22 (46.8%) to 51 (79.7%), and that of enterotoxin type A strains from 27 (57.4%) in 1991 to 47 (73.4%) in 1992. The number of strains which produce Toxic Shock Syndrome Toxin-1 (TSST-1) also increased from 19 (40.4%) to 45 (70.3%), and those strains that produce β-lactamase decreased from 24 (51.1%) to 21 (32.8%). From the above results, we confirmed the recent change in types of the epidemic strains of MRSA. Namely, there was a marked increase in number of strains which produce type II coagulase type A enterotoxin and TSST-1. For the prevention of a patient to patient-, room to room-and ward to ward-spread, strict isolation was indicated both the infected patients, immunocompromised patients who were at high risk for the infection and the proved carriers. Treatment with VCM was started immediately if MRSA infection was thought plausible. These countermeasures seemed to succeed in reducing the incidence of the infection in our ward.

View full abstract

The in vitro activity of a murine monoclonal antibody (E5) reactive with endotoxin was examined in human whole blood by measuring the luminol-chemiluminescence (CL) activity in response to phorbol myristate acetate (PMA) as an index of the priming effect of lipopolysaccharide (LPS) on the release of reactive oxygen species. Whole blood samples obtained from healthy adults showed a significantly enhanced CL response to PMA after incubation with LPS (100 ng/ml, Escherichia coli O111: B4) for 10 min at 37 & deg;C, as compared with untreated blood samples, though no CL response was induced by LPS itself. This priming effect of LPS varied from person to person. Similarly, various degrees of the priming effect were observed with other LPS preparations derived from E. coli O55: B5, Klebsiella pneumoniae, Serratia marcescens and Salmonella typhimurium. However, the priming effects of these LPS or a synthetic lipid A (LA-15-PP) of E. coli were significantly prevented to various degrees when such endotoxins were treated with E5 for 30 min at 37 & deg;C prior to being added to blood samples. The inhibitory effect E5 was dosedependent and was most potent against the LPS of E. coli O111: B4.
These results indicate that E5 suppresses the priming effect of LPS on oxygen radical release from human whole blood, and therefore suggest that E5 may be a useful drug for supportive therapy in patients with gram-negative septicemia or endotoxemia, especially in a case involving serious neutrophil-mediated organ injury caused by excessive release of oxygen free radicals.

View full abstract

Using a severely complicated in vitro bladder model with a computer-controlled autosimulationsystem for antimicrobial concentration in urine, we examined the significance of the peak concentration value and time above MIC, and obtained the following results.
1. Regarding the initial bactericidal speed, the higher the peak concentration value was, the more quickly the bacterial concentration decreased.
2. Concerning the minimum value of bacterial concentration in the model bladder, the higher the peak concentration value was, the more the level tended to decrease.
3. Regarding inhibition of regrowth of bacteria (ERT)
1) When a particular antimicrobial agent had no PAE on bacteria, the viable bacterial count curve was similar in cases of peak antimicrobial concentration values of higher than 8 MIC.
2) On the other hand, when an antimicrobial agent had PAE on bacteria, regrowth of bacteria occurred quickly below a certain peak concentration (24 MIC or 64 MIC value), although there was some difference depending on the type of antimicrobial agent. Thus, it was suggested that the peak concentration above that level be attained and maintration above that level be attained and maintained for some time were in order to obtain PAE adequately.
4. It is concluded from the aforementioned facts that two factors, a certain high concentration and a certain continued time are necessary for both antimicrobial agent with metabolic disorder and antimicrobial agent with cell wall synthetic inhibition to obtain proper antimicrobial effect. The condition of their necessity differed slightly according to the two different agents.

View full abstract

We experimentally investigated an efficient administrating method of antimicribials prior to the clinical treatment of complicated urinary tract infection without catheter. An exerimental model of moderately complicated urinary tract infection, which can simulate changes in the urinary concentration of antimicrobials by means of previously reported computer control method, was used for the experiment.
The following results were obtained.
1. At first, for complicated UTI cases without an indwelling catheter, an investigation was made of the clinical therapy results to determine the upper MIC range of bacteria, including isolated strains, that can be nearly eliminated following treatment, by the administration method. We then estimated the clinical breakpoint.
2. By adjusting the size of the diverticulum in a urinary bladder model, we prepared a model which gave comparatively good agreament between the clinical and experimental breakpoints.
3. The clinical therapy results revealed that by the clinical breakpoint was higher in degree of coverage of the MIC distribution of causative bacteria associated with a higher actual clinical elimination rate and a higher efficacy rate.
4. It was surmised that an efficient administration method for complicated UTI without an indwelling catheter can be established if the urinary bladder model and the following approach are applied: prior to clinical treatment it should be determined what would be high enough to cover 70% of the MIC distribution of the estimated causative bacteria of UTI, and then an administration method which would yield that concentration as the experimental breakpoint should be devised.

View full abstract

For the purpose of conducting a therapeutic study on biofilm of the urinary tract, we devised a computer-controlled severely complicated bladder model (biofilm model of the urinary tract) enabling us to simulate the time-course of the concentration of antimicrobial agents in the urine. Using this model, we investigated clarithromycin (CAM), which has been reported to have anti-biofilm action, at concentrations close to its urinary levels at the time of clinical use in order to predict its effect on biofilm of the urinary tract. On the basis of those experimental results, we also conducted a clinical examination. The following results were obtained.
1. The action of ciprofloxacin (CPFX, MIC: 8μg/ml) alone, which shows anti-P. aeruginosa activity, caused apparent elimination of P. aeruginosa from the model. However, regrowth of the microbes occurred when CPFX was removed from the bladder model. Moreover, the biofilm was not eliminated by the antimicrobial action of CPFX, and this was surmised to be the cause of the regrowth.
2. CAM (MIC: above 128μg/mi), which has no anti-P. aeruginosa activity, was similarly tested as anti-biofilm agent when added alone to the biofilm model. The P. aeruginosa recovered to its initial concentration within 48 hours, but the biofilm disappeared due to the action of CAM.
3. The combined action of CPFX and CAM caused microbial elimination from the bladder model without microbial regrowth, even after these antimicrobial agents were removed from the bladder model. After the action of CPFX and CAM, the biofilm disappeared, and no microbial adherence was noted.
4. Measurement of time-course of the alginate content, which is the main component of P. aeruginosa biofilm, in the presence of CAM found that the alginate content decreased below the limit of detection after day 5.
5. The clinical study of complicated urinary tract infection revealed the microbial elimination rate and the efficacy rate to be higher in the combined CPFX-CAM administration group than in the CPFX-only administration group.
6. Based on the above results, we surmise that the combined use of an antimicrobial agent which is active against the causative microbe and anti-biofilm agent such as CAM will show some degree of efficacy in eliminating biofilm of the urinary tract.

View full abstract

A 44-year-old male, who had been to Lagos, Nigeria, was admitted to our hospital because of a high grade fever on July 20, 1993. On admission, Plasmodium falciparum was detected in his blood smears and the antibody titers against P. falciparum andPlasmodium vivax antigens were 1: 256 and <1: 4 respectively by the indirect fluorescent antibody test. Therefore, he was diagnosed as having P.falciparum malaria. He was treated with halofantrine (Half anTM: Smith Kline Beecham Pharmaceuticals, England), two tablets at six-hourly intervals, a total of six tablets (1500 mg). Parasites were cleared rapidly and remission was achieved without any adverse reactions. Halofantrine can therefore be recommended for the treatment of imported P. falciparum malaria.

View full abstract

The patient was a 26-year-old man who complained of headache and vomiting. On examination, there was nothing abnormal, but the edge of the right optic papilla was not clear. His temperature was 38.5°C, pulse 96/min, blood pressure 120/80 mmHg. A space-occupying lesion in his fronto-dextra cupular part was found by CT scanning. He had a 12-year history of chronic purulent otitis. The diagnosis was a brain abscess in the fronto-dextra cupular part. The brain abscess was extracted and Pasteurella multocida was isolated from the dark brown pus draining from the abscess. The patient recovered through proper antibiotic therapy based on a sensitivity test. Reports of infections caused by this organism in foreign countries very widely from local infections due to bites and scratches by cats, dogs etc: to general infections such as infections of the respiratory tract, sepsis and meningitis. However, Pasteurella multocida brain abscesses are rare.
Pasteurella multocida is a Gram-negative short rod which is best known as part of the mouth flora and as a pathogen causing septicemia in many domestic animals, such as cats, dogs etc. Infection in man results mainly from animal bites or scratches. It has been reported that Pasteurella multodida can cause human septicemia, meningitis, respiratory tract infection, conjunctivitis and other infections. We isolated a strain of Pasteurella multocida from the pus of a brain abscess following chronic purulent otitis on August 6, 1990.

View full abstract

We diagnosed a 41-year-old female patient to be suffering from Chlamydia pneumoniae (C.pneumoniae) by using PCR and culture methods. She had a prolonged dry cough and slight fever. Her chest roentgenogram showed a segmental infiltration in the middle of the right lung field. We treated her with 400 mg of cefpodoxime proxetil (CPDX-PR) per day. On the 4th day after beginning the treatment with CPDX-PR, she still complained of a productive cough. We changed the treatment by using 300 mg of roxithromycin per day and these symptoms disappeared.
To diagnose C. pneumoniae early, PCR, MIF and culture methods are very useful diagnostic tools.

View full abstract

We report a case of Listeria meningitis associated with systemic lupus erythematosus (SLE). The case is a 29-year-old female who was diagnosed as SLE on August, 1982, and had been followed since then. On May 25, 1993, she was admitted to our hospital with complaints of fever, head ache and vomiting which developed 3 days before admission. An examination of, the cerebro-spinal fluid (CSF) revealed a cell count of (3664/3/mm³), protein concentration (123 mg/dl), and the quotient of CSF (Q albumin) (27.5) were elevated. A diagnosis of meningitis was made and therapy of antibiotics was begun. A CSF culture on admission was positive for Listeria monocytogenes, but the blood culture was negative. Clinical symptoms disappeared in about a week and abnormal CSF findings returned to normal in about a month with the administration of antibiotics. Listeria infection is a rare disease, but sometimes develops as an opportunistic infection in immunocompromised hosts. As far as we know, only 13 cases have been reported so far (9 cases in foreign countries, 4 cases in Japan).
We also discuss Listeria infections associsted with SLE referring to the former reports.

View full abstract

Two cases of lung infection due to Mycobacterium chelonae subsp. abscessus are reported. Case 1, a 50-year-old female, was a secondary infeciton-type, and case 2, a 53-year-old female, was a primary infection-type. The 16 cases reported, between the ages of 29 and 76 years, there were 6 males and 10 females. The roentgenographic examinations, revealed that the ratio of the primary and secondary infection-type was 3: 1. Effective agents for this organism has not been yet confirmed. In the presaent study, we treated two patients with AMK and IPM/CS, and obtained negative conversion of the sputum culture as well as improvement of roentgenographic features.

View full abstract