AZT susceptibility of sequential 12 HIV-1 isolates was investigated by culture method. These isolate were obtained from three patients who had received AZT for at least 12 months and switched to ddI after they appeared to deteriorate clinically. Cultures of isolate from a patient before therapy with AZT or ddI did not show cytopathic effect (CPE) in the presence of 0.1 μM or 1.0μM AZT. Culture of 2 isolates from two patients treated with AZT more than 6 months before switched to ddI therapy showed CPE in the presence of AZT and HIV-1 p24 antigen was detected by ELISA in the supernatants. Culture of 6 of 7 isolates from three patients treated with ddI after long-term therapywith AZT also showed CPE in the presence of 1.0 μM AZT and HIV-1 p24 antigen was detected. These results demonstrated that AZT-resistant variant was still a dominant population in the isolate from patients after 11 to 13 months of discontinuing AZT therapy.

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We determined whether black tea extract inhibits the infectivity of influenza virus to mice. When mice were inoculated intranasally with 10^5.3 PFU influenza viruses (10^1.3 LD₅₀), their body weight decreased and all died within 10 days. Whereas, when mice were inoculated i.n. with the mixture of influenza viruses and 2%(w/w) black tea extract, 5 min after mixing, all mice showed normal bodyweight increase and survived. Neutralizing antibody to influenza virus was not detected in nine of 10 survived mice.
The results indicate that black tea extract at beverage concentration (2% w/w) inhibits almost completely the infectivity of influenza virus to mice and that in vivo reversion of the tea-inactivated influenza virus does not occur.

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We report four cases of infective endocarditis due to nutritionally variant streptococci (NVS) that occurred between 1981 and 1991. Three female and one male patients had underlying heart diseases. Causative organisms showed satellitism to staphylococci. Two strains were identified as S. adjacens and the other two were identified as S. defectivus by DNA-DNA hybridization. All strains had tolerance and one strain had resistance to benzylpenicillin (MIC 4 μg/ml). This penicillin-resistant strain also had tolerance to gentamicin. There was no synergism of benzylpenicillin and gentamicin against this strain by a killing curve in vitro. A 11-year-old female patient with infective endocarditis due to this strain, who had a transposition of great arteries, had large vegetations in external conduits by the Rastelli's operation. Some intensive antimicrobial chemotherapies were unsucessful and a surgical replacement of the conduits must be done in this case. Since infective endocarditis due to NVS is not rare in Japan, NVS should be considered for the causative organisms in culture negative endocarditis.

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Postmortem infectious lesions were analyzed in 63 patients with bacteremia and fungemia. Bacterial infection was found in 36 patients, deep mycoses in 27 and cytomegalovirus infection in 7. Among deep mycoses patients, yeast was noticed in 17, Aspergillus in 13 and Mucor in one. Infectious lesions were not observed in 10 cases. Fifteen cases of 23 leucopenic patients were complicated with deep mycoses.
Deep mycoses was noticed in 43% of bacteremia and fungemia patients, but not in candicemia patients. Fungemia due to Candida was related to blood access, however, not to deep mycoses. On the other hand, disseminated mycoses was found in 4 of 5 cases with Trichosporon beigelii fungemia. T. beigelii infection is noticeably life-threatening to the immunocompromised host.

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Changes in susceptibilities to oral antimicrobials were investigated in 189 children, in whom Haemophilus influenzae was detected by nasopharyngeal culture, among the children who were diagnosed as having acute otitis media or chronic sinusitis with acute exacerbation in the Department of Otolaryngology in 1986, 1988 and 1991. The minimum 50% inhibitory concentration (MIC50) and minimum 90% inhibitory concentration (MIC90) of ampicillin (ABPC), cefaclor (CCL) and erythromy cin (EM) in the subjects examined in 1986 were almost the same as the levels in those examined in 1991. The MIC50 level of minocyclin (MINO) in the subjects examined in 1991 was lower by 2 testtubes than that determined in 1986, showing improvement in susceptibilities. The prevalence of resistance to antimicrobials was compared between 1986 and 1991. The prevalence to ABPC tended to decrease very slightly, while the prevalence to CCL tended to increase very slightly. The prevalence of EM tended to increase slightly. Many CCL-resistant strains detected in 1991 showed resistance to ABPC and EM, but showed favorable susceptibilities to cefixime and norfloxacin. Future trends of CCL-resistant strains should be carefully observed. There was no marked change in the prevalence of resistance to ABPC or CCL. ABPC and CCL were considered to be the first choice drugs, even at present, for pediatric patients with acute otitis media due to infection with H. influenzae and in those with acute exacerbation of chronic sinusitis due to the infection.

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A new animal model of recurrent pulmonary aspergillosis was established using cyclophosphamide (CPM) instead of cortisone acetate for reconstitution of immunosuppression, and histopathological findings were compared with those of the previous model.
A recurrence of pulmonary aspergillosis, according to granulocytopenic conditions was much more rapid and uniform in this model than that of steroid induced model shown. In histopathologically, invasive and massive proliferation of Aspergillus hyphae was observed in lungs of rats 5-7 days after the CPM treatment and hyphal invasion into pulmonary vessels which suggested rapid systemic spreading of the infection was also found.
This model considered to be good for the simulation of recurrent aspergillosis in patients with severe immunosuppression and also useful for studies of new diagnostic and therapeutic methods of recurrent pulmonary aspergillosis.

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To determine the efficacy of an additional low dose of intradermal injection of recombinant HB vaccine administered to nonresponders in addition to the standard three injections, we investigated the and-HBs positive rate and titer of and-HBs after four years among 58 subjects who did not develop antibodies after three doses of subcutaneous injections and who did develop the antibody after an additional low dose intradermal injection (Nonresponders), and compared them with 150 subjects who developed anti-HBs after three doses of subcutaneus injections (Responders). Fifty four subjects who were negative after four years were revaccinated with one or three doses of the subcutaneous injection.
Anti-HBs positive rates after four years were 56.0 percent in Responders and 56.9 percent in Nonresponders, showing no differences. Eighty eight percent of 54 subjects who were given one or three doses of the vaccine developed anti-HBs. There were no differences in the antibody rates or titer of the antibody between the one and three doses groups.
The decrease in titer of antibody was related to the levels initially having the most marked decrease in the level four years later.
From these findings, we concluded that there were no differences of and-HBs positive rates or antibody titer after four years in Responders and Nonresponders, and that one additional dose of the vaccine may be effective four years after initial administration among both groups.

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An experimental ascending urinary tract infection was induced by transurethral instillation of Escherichia coli in streptozotocin (STZ)-induced diabetic mice, in which bactericidal capacity of the perineal exudating neutrophils had been damaged. The local immune response inuninfected diabetic mice and the response time fluctuation of local immune responses in diabetic mice following infection were compared with those in normal mice, and the nature of induced changes inthe local immune response was determined.
The diabetic model was prepared by a single administration of 280 mg/kg of STZ after it had been fasted for 15 hours. The mouse was then fasted an additional 2 hours to induce the diabetic state. Compared with the normal mice, the diabetic mice prepared using this method were found to have a significant suppression of the bactericidal capacity of the perineal exudating neutrophils, 3 weeks after induction of diabetes.
The study of the cellular distribution in uninfected urinary tract tissue demonstrated a reduction in CD4-positive T cell distribution in bladder submucosa of diabetic mice at 3 weeks after induction of diabetes (diabetic group), compared with that in the normal mice (control group). In the diabetic group, probably in order to compensate for the reduction in CD4-positice T cell distribution in the bladder submucosa, the distribution of macrophages was increased in the bladder epithelium, compared with the control group. This finding suggested that diabetic mice are thought to have protective mechanisms against infection different from those of normal mice in the uninfected state.
The study of the response time fluctuation of local immune response at the infected sites demonstrated infiltration of macrophages was the same grade as that of neutrophils in the control and the diabetic group. However, in the diabetic group, a marked infiltration of macrophages was recognized when compared with the control group in the early phase of infection. These findings suggested that macrophages aid in protection against infection when damage tothe bactericidal capacity of neutrophils results in insufficient elimination of microorganisms. On the other hand, infiltration of T and B cells was weaker in the diabetic group, than in the control group.

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We conducted a non-randomized, prospective study comparing the efficacy of tosufloxacin with that of norfloxacin in preventing bacterial infection during chemotherapy-induced neutropenic episode. Fifty-one patients with hematological malignancies were included, and a total of 108 episodes of neutropenia were studied. There was no significant difference between the two groups in the incidence of fever, in the time-to-event analysis of febrile episodes nor in the incidence of gram-positive bacteremia despite the broader spectrum of tosufloxacin. The incidence of gram-negative bacteremia was lower in patients treated with the quinolones as a prophylactic than in the historical controls given polymyxin B. Especially the incidence of bacteremia due to Pseudomonas aeruginosa was significantly less when tosufloxacin was used. A possible synergistic effect of sulfamethoxazole-trimethoprim in preventing the febrile episode was also observed.

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Candida is present in the flora of the oral cavity, skin, intestinal tract and vagina, and is also known to be an opportunistic pathogen. Infection with this fungus has been increasing annually along with wide spread use of broad-spectrum antimicrobial agents.
The subjects included 95 patients (48 males and 47 females) who had been diagnosed as having had deep-seated candidiasis, amoung patients autopsied between 1982 to 1991. In regard to annual changes in deep-seated candidiasis, the incidence reached a peak in the 1985 to 1988 period, and thereafter decreased. The number of cases with leukemia as the underlying disease was the largest, 36 (37.9%), followed by malignant lymphoma in 10, and aplastic anemia 5. The number of cases with infection of the stomach was largest, 42 (44.2%), followed by the esophagus in 33 (34.7%), the lung and kidney.
The cases with deep-seated candidiasis showed low values of or level of lymphocyte, hemoglobin, CRP, total protein and cholesterol and high values or levels of LDH, urea N, creatinine and total bilirubin. Cases with marked decrease in neutrophils showed no regional infiltration of inflammatory cells in any of the organs infected with Candida. Cases with disseminated canditiasis showed vascular invasion by Candida. The laboratory findings also showed that most of the cases had been undernourished and had high values of CRP which supports the presence of inflammation. Common sites of infection are the esophagus, stomach, and intestinal tract. In the presence of granulocytopenia and immunodeficiency, tissue invasion become severe and associated with vascular invasion.

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To make an early diagnosis and decision for prompt treatment of the children with tuberculosis, we analyzed 61 cases in both inpatients and outpatients for the clinical and laboratory characteristics.
One forth of the 61 cases were below 1-year-old, and 43 cases (70.5%) had a positive family history. The tuberculin test revealed that all of the 61 cases were positive, and 18 of them had a strongly positive reaction (≥30 mm). Although the serum levels of inflammatory markers including white blood cells counts, erythrocyte sedimentation rate and CRP value increased only slightly in most cases, serum IgM levels in 59.0% of the children with tuberculosis were twice to three times higher than those in normal average levels. Especially in 96.2% of the children hospitalized for the need of isolation the IgM levels were demonstrated high.
Thus, in children with tuberculosis the detection of serum IgM levels as well as family history, tuberculin reaction, and chest X-ray findings may be helpful for the early diagnosis and prompt decision of hospitalization.

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With the view of making an adaptation study for biofilm of the urinary tract, we devised a computer-controlled highly complicated bladder model (biofilm model of the urinary tract) that allowed the simulation of time-course changes in the urinary concentration of antimicrobial agents.
Clarithromycin (CAM), which is reported to have an anti-biofilm action, was examined at urinary levels approximating clinical concentrations and its effect on biofilm was determined.
The following results were obtained.
1) Ofloxacin (OFLX, 200mg×2/day, MIC; 8μg/ml), which is active against Pseudomonas aruginosa, caused apparent microbial elimination from the model at 42 hours, but bacterial regrowth occurred 4 hours after withdrawal of this agent. No disappearance of the biofilm was noted with OFLX suggesting that this was the cause of bacterial regrowth.
2) The combination of OFLX (200 mg×2/day) and CAM (MIC;>128μg/ml, 200 mg×2/day) on anti-biofilm agent, with no effect on P. aeruginosa, eliminated bacteria from the bladder model more rapidly and prolonged the regrowth time to 10 hours after withdrawal of the antimicrobial agents. Disappearance of most of the biofilm and only slight microbial adhesion was noted.
3) The combination of OFLX (200 mg×2/day) and CAM (400 mg×3/day) caused microbial elimination from the bladder model with no regrowth at 30 hours after withdarawal of the antimicrobial agents. The biofilm disappeared completedly and no microbial adhesion was noted.
4) CAM alone (400 mg×3/day) allowed microbial recovery to the initial level within 48 hours after withdrawal, but led to disappearance of the biofilm and the adhesion of microbes without a glycocalyx.
5) These results suggest that the anti-biofilm action of CAM is dose-dependent, and that combined use of an appropriate antimicrobial agent and anti-biofilmagent like CAM may be effective for biofilm infections of the urinary tract.

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Intractable complicated urinary tract infections (UTI) are caused by host and/or bacterial factors which predispose to persistent infections and recurrent infections. It is still unknown what kind of factors are responsible for the intractable complicated UTI. With the increase of compromised host or cases with complicated UTI, the factors involved in intractable and recurrent UTI are diversified. Accurate diagnosis of the factors affecting the therapeutic effect will have more importance.
The factors affecting the therapeutic effect was subjected to multiple regression analysis from both aspects of underlying diseases in the urinary tract (complicated factors) and systemic conditions (compromised factors) in one hundred and ninety patients of complicated UTI admitted to our clinic.
Sex, presence or absence of hydronephrosis and indwelling catheter and volume of residual urine as complicated factors and age, serum creatine value, peripheral neutrophil count, peripheral lymphocyte count, diabetic or not, whether the patient underwent major operation within 1 week or not, and serum albumin value, an indicator of malnutrition, as compromised factors were analyzed by multiple regression analysis.
The presence of indwelling catheter, residual urine more than 50 ml and hypoalbuminenia less than 3g/dl were the most determinant of the clinical efficacy in cases with complicated UTI. Interestingly, presence of residual urine more than 50 ml is considered an equally intractable factor with the presence of indwelling catheter. These factors proved to be important also as recurrent factors. In the host with these factors, UTI are often caused by resistant bacteria and indicated to be intractable also bacteriologically.

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A new immunochromatography assay technique (DAINASCREEN HBsAg, Dainabot) has been recently introduced for the detection of the presence of hepatitis B surface antigen (HBsAg). To evaluate the feasibility of using the DAINASCREEN HBsAg, we carried out comparison tests with this method and RPHA.
Results obtained were as follows:
In a test of 692 sera from inhabitants in Iki island, Nagasaki Prefecture, 24 (3.5%) were positive for HBsAg by both DAINA and RPHA. Twenty-four patients with hepatitis B were all positive in both methods. Thirty patients with autoimune diseases and eight who had accidental needle exposurse to the blood of patients with HBV infection were negative by both methods.
Six individuals were followed over a period of 10 years and all eventually climinated HBsAg from their sera. In four patients, HBsAg remained positive by DAINA at one yearly check after it had become negative by RPHA. In five patients, HBsAg remained positive by RIA and IMs after DAINA and RPHA had become negative.
A good correlation was observed between DAINA and RPHA with a concordance rate of 99.5%. A good correlation of HBsAg titer between the two methods was observed and found to be significant (r=0.87, p<0.001).
HBsAg assay was able to be completed within 15 minutes by DAINA and the procedure was simple.
These results indicate that the sensitivity of DAINA is superior to RPHA and it is easy to use.

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We developed an improved, highly sensitive non-radioisotopic (non-RI) reverse transcriptase (RT) assay (RTA). While the original non-RI method previously reported made use of primer immobilization, our improved method was based on a primer-template immobilization procedure. We tested the template specificity, reproducibility and linearity of the new method in assays of human immunodeficiency virustype-1 HIV-1 RT. The sensitivities of the method previously reported, the improved method and the sensitive radioisotopic (RI-) RTA were compared in assays of recombinant HIV-1 RT, partially purified HIV-1 particles, and the culture supernatant derived from HIV-1-infected cells. For each of these samples except the culture supernatant the improved method was the most sensitive. It appeared that the fetal bovine serum presented in the culture medium interfered with the assay reaction. The curve describing inhibition of the assay reaction by fetal bovine serum showed that the highest degree of sensitivity in the assay was obtained when the culture supernatant sample was diluted four times. With this degree of dilution, the sensitivity of the new method for assay of culture supernatant sample was still half that of the sensitive RI-RTA. Culture supernatants of five peripheral blood mononuclear cell samples obtained from HIV-1-seropositive carriers were assayed by both the improved method and the sensitive RI-RTA; and with each of the methods, however all virus-positive cultures could be detected. The improved non-RI RTA was considered especially useful for assay of culture supernatants for purposes of virus isolation because of its advantages of excellent sensitivity and lack of requirement for radioisotopes.

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A 51-year-old male was admitted because of cough and high fever. His chest X-ray and CT revealed milialy shadows. he was administrated some antibiotics. But on the 8th day, his chest X-ray revealed ground glass shadows and he was under severe hypoxemia. We suspected milialy tuberculosis with acute respiratory failure due to his clinical course. The intensive therapy with antituberculosis drugs, hydrocortisone (1 g/day) was started and intubation was performed. The smear of the clinical samples did not show acid-fast bacilli on admission. However, by the polymerase chain reaction amplification method specific DNA fragments of Mycobacterium tuberculosis complex were detected from BALF and bone marrow. He was extubated after the 13th day and he recovered. It is difficult to separate and identify tubercle bacilli rapidly, but PCR is available for early diagnosis.

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A 32-year-old woman was hospitalized with the chief complaints of high fever and right flank pain. The patient had received treatment for diabetes mellitus and liver cirrhosis. The patient's laboratory data indicated pyuria, renal dysfunction and hyperglycemia. E. coli was detected in the blood, urine and pus. Plain abdominal X-ray revealed gas shadows at the right renal region. Abdominal CT scanning also showed gas shadows in the renal parenchyma of both sides. A diagnosis of bilateral emphysematous pyelonephritis was made. Chemotherapy and retroperitoneal drainage was performed. After therapy, the patient's laboratory data was improved and the abnormal gas shadows disappeared. We reviewed 77 cases of emphysematous pyelonephritis, including our case, from the Japanese literature.

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A 24-year-old Japanese male who had traveled to India was referred to our institute because of severe prolonged watery diarrhea. Cryptosporidium parvum oocysts were detected in his stool specimens by the phase-contrast microscopy and modified acid-fast stain. Stool examinations were negative for other intestinal protozoas. Bacterial cultures were also negative for pathogenic agents. He was treated with sulfamethoxazole/trimethoprim (Co-trimoxazole) for 6 days and he soon recovered.
To our knowledge, there have been no reported cases concerning imported cryptosporidiosis in Japan. Physicians should regard this organism as one of the causative agents of travelers' diarrhea when other pathogens are negative, since this organism is easily overlooked in routine stool examinations.

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