Inhibition of pulmonary metastasis by Nocardia rubra cell wall skeleton, with special reference to macrophage activation.

The antimetastatic activity of Nocardia rubra cell wall skeleton (N-CWS) with or without cyclophosphamide was examined in an experimental model of pulmonary metastasis induced by Lewis lung carcinoma in C57BL/6 mice. Lewis lung carcinoma cells were implanted into the footpads of mice, and the implanted tumors were removed 9 to 10 days later. Pulmonary metastatic nodules began to develop a few days after the implanted tumor was removed. The inhibitory effect of N-CWS was evaluated from the number of pulmonary surface nodules about 3 weeks after tumor implantation. The antimetastatic activity of N-CWS depended upon the dose, time, and route of its injection. Injection of N-CWS i.v. after removal of the implanted tumor caused the greatest inhibition of development of pulmonary metastases. Therapy with N-CWS plus cyclophosphamide prolonged significantly the survival of mice with metastases. The cytotoxic activities of peritoneal macrophages and macrophages in the lung against Lewis lung carcinoma cells were enhanced in mice treated with N-CWS. Injection i.v. of peritoneal macrophages activated with N-CWS inhibited pulmonary metastases. The role of macrophages in inhibition of micrometastasis in the lung is discussed.

Production of a tumor cytolytic factor(s) by activated human alveolar macrophages and its action.

When human alveolar macrophages (AM) lavaged from healthy donors were incubated in medium with or without lipopolysaccharide (LPS) or muramyl dipeptide, they released a factor(s) responsible for tumor cell killing. The activity of the tumor cytolytic factor(s), called TCF, was determined by radioactive release assay. Human AM released variable amounts of TCF into the culture medium without any stimulation, but the release was stimulated significantly by LPS (0.1 micrograms/ml) or muramyl dipeptide (1 micrograms/ml). Maximal production of TCF by the AM was detected in the supernatant after treatment for 3 hr with LPS, and the extent of TCF release correlated with the density of AM. In cultures with LPS, the ability of activated AM to secrete TCF was maintained for 48 hr but was lost by 96 hr. After its loss, the ability to produce TCF could be restored by a second treatment with LPS. Full expression of lysis by TCF to lyse tumor cells required its interaction with tumor cells for at least 24 hr. TCF destroyed human allogeneic tumor cell lines but did not affect nonneoplastic cell lines. TCF activity was resistant to treatment with protease inhibitors, superoxide dismutase, or catalase and to heating at 70 degrees for 1 hr, but it was labile on heating at 100 degrees for 10 min. The tumoricidal activity in the supernatant of activated human AM indicates a potential effector mechanism by which AM kill neoplastic cells.

In vitro generation of tumoricidal properties in human alveolar macrophages following interaction with endotoxin.

Human alveolar macrophages (AM) obtained by bronchoalveolar lavage from healthy nonsmoking donors exhibited primarily low levels of cytolytic activity against allogeneic tumor target cells. These AM acquired enhanced capacity to kill tumor cells following a 24-hr incubation in vitro with endotoxin [lipopolysaccharide (LPS)]. Maximal tumoricidal activity of LPS-activated AM as measured by lysis of tumor target cells was obtained after incubation with tumor cells for 72 hr. LPS-activated AM lysed allogeneic tumor cell lines of different origins but did not affect normal, nonneoplastic cells. We conclude that LPS induces human AM to become tumoricidal. This method should be useful in studies on therapeutic agents enhancing AM-mediated cytotoxicity in situ.

[Report of questionnaire about enterohemorrhagic Escherichia coli cases caused in the area including Sakai City in 1996].

Many outbreaks of EHEC O157 were occurred in Japan in 1996. There was the biggest outbreak involving approximately 6000 victims in area of Sakai City. We conducted a questionnaire survey on the EHEC infection to the physicians who treated these patients and the microbiologists. For physicians we asked mainly clinical manifestation of patients they observed and treatment they did and their outcome (Answer A). Microbiologists were also required to answer the opinion about treatment (Answer B). Eighty-five of physicians replied about their 222 patients and 209 of microbiologists answered their opinions. In this report we summarized the data and obtained following results: 1) We observed higher frequency of HUS/TTA complication in female than male. 2) Higher HUS/TTP complication was accompanied with patients complained symptoms of fever, bloody diarrhea, and general fatigue. 3) The most patients (96%) were received antimicrobial agents, especially fosfomycin (84%), followed by new quinolon (17.8%) and Cefems (12.2%). Later starting of treatment with antimicrobial agents than 7 days from onset of symptoms increased HUS/TTP complication. 4) Anti-diarrhoeal agents seems to be one of risk factors for HUS/TTP complications.

[Studies on erythromycin stearate capsules (OE-7) (author's transl)].

The blood concentration and urinary excretion of OE-7 (erythromycin stearate capsules) and control drug were investigated in 6 volunteers having gastroptosis. OE-7, newly arranged capsules containing erythromycin stearate, was investigated and obtained the results of high blood concentration by oral administration. To confirm the above, we measured blood concentration and urinary excretion of OE-7 comparing with ordinary erythromycin stearate capsules in 6 volunteers having gastroptosis. The peaks of blood concentration were noted at 3 hours after administration in 6 volunteers uniformly. The mean maximum blood concentration of OE-7 was 1.17 mcg/ml which was significantly higher than ordinary erythromycin stearate capsules. In time course of mean blood concentration, the blood concentration levels of OE-7 were higher than those of control erythromycin stearate capsules at any measurement. Effective blood levels were continuously high in OE-7. Urinary excretion of OE-7 reached to the maximum from 4 to 6 hours after administration which was also higher than control erythromycin stearate. Clinical efficacy of OE-7 was investigated in 28 cases in acute respiratory infection. The results noted were excellent in 5 cases (17.9%), good in 16 cases (57.1%), no change in 6 cases (21.4%), and undetermined in 1 case (3.6%). There were 5 cases of slight gastro-intestinal discomfort as the side effects. As the conclusion, OE-7 revealed good bioavailability and seemed to be useful antibiotic for acute respiratory infections.

[Blood concentration and urinary excretion in human of OE-7, a gastric hard capsule containing enteric-coated granules. I (author's transl)].

The investigation of physico-chemical properties and measurement of blood concentration and amount of excretion into urine in human of OE-7 (main constituent is erythromycin stearate) were performed as the fundamental studies. The results obtained were as follows; i) OE-7 is a gastric hard capsule containing enteric coated granules. ii) The contents of OE-7 capsule (granules) were resistant to acid and was favarable in the enteric solubility. iii) In comparison with the conventional erythromycin stearate capsule in healthy adults, OE-7 showed a relatively small individual difference in blood concentration after administration. iv) OE-7 was higher in the mean blood concentration and was superior in the persistence of effective blood concentration compared with the conventional erythromycin stearate capsules.

[Clinical studies on cephradine (CED) (author's transl)].

Cephradine (CED) was administered intravenously to the patients with respiratory infection or urinary tract infection with the dosage of 1.0 approximately 4.0 g daily and their clinical effects were investigated. Good clinical effects of CED were obtained in 5 of 8 cases with respiratory infections and in 2 of 5 cases with urinary tract infections. Side effects were noted in 2 cases, but they improved rapidly by the cessation of the drug administration. One case developed skin eruption and the other case complained general burning, palpitation after intravenous injection and slight insomnia. CED is seemed to be effective antibiotic for the respiratory as well as urinary tract infections clinically. However it was advisable that careful attention should be paid when CED is administered to the patient having drug allergy or nephropathy.

[Clinical study of miconazole in deep-seated fungal infections].

We administered miconazole to patients with deep-seated fungal infections, to investigate into the clinical and mycological responses to the drug and also into its clinical safety. We also looked into the responses of amphotericin B (AMPH), flucytosine (5-FC) and the combination of the 2 drugs to deep mycoses in the past 10 years, for retrospective comparison of the findings achieved in the treatment with miconazole. A clinical response rate of 85% (29/34 patients) was achieved in the treatment of deep-seated fungal infections with miconazole. Mycologically, a fungus eradication rate of 79% (22/28 patients) and a fungus decrease rate of 11% (3/28) were achieved with the miconazole treatment, comparison of the response to miconazole alone and that to AMPH alone revealed that the former was significantly preferable. Side effects of miconazole were observed in 23% of the treated patients (15/66). Statistical analysis of the incidence of side effects of miconazole and that of AMPH showed that the former was significantly lower.

[Clinical effect of miconazole gel against upper digestive tract mycosis].

A total of 43 patients, comprising 41 patients with oral candidiasis and 2 with esophageal candidiasis, were treated with miconazole (MCZ) gel to assess its efficacy and safety in treating upper digestive tract mycosis. The efficacy of the drug was evaluable in 33 of them, consisting of 32 patients with oral candidiasis and 1 with esophageal candidiasis. The clinical efficacy rate of the drug against oral candidiasis was 87.5% (28/32 patients), and the clinical response was good in the 1 evaluable patient with esophageal candidiasis. The safety of the drug was assessed in 40 patients. In 3 (7.5%) of them, nausea occurred as an adverse event, but was not particularly serious in any of them. No abnormal laboratory test values caused by the drug were observed. The results suggested that MCZ gel would be a very useful drug in treating oral and esophageal candidiasis.

[Multicenter clinical trial of itraconazole in the treatment of pulmonary aspergilloma. Pulmonary Aspergilloma Study Group].

Itraconazole 100-200 mg was orally administered to 49 patients with pulmonary aspergilloma once a day in principle immediately after breakfast. Drug concentrations in fungus balls and in aspergilloma cavities were determined in each measurable case and the data were evaluated in terms of efficacy and safety. Overall evaluation was done considering the following 3 criteria: improvement of clinical symptoms, X-ray findings and mycological findings. The efficacy rate was 63.4% (26/41 cases). Out of 49 cases, adverse reactions and abnormal laboratory test values were observed in 8 (16.3%) and 13 (26.5%) cases, respectively. The safety rate was 81.6% (40/49 cases). 160 ng/g-4, 010 ng/g of itraconazole was detected in aspergilloma in 4 measurable cases. This finding showed that itraconazole had infiltrated into the fungus balls. Itraconazole was detected in aspergilloma cavities as well: drug concentration was 825 ng/g and 1,020 ng/g respectively. Based on overall view of clinical effects and drug concentrations in fungus balls and in the walls of lung cavities, it is concluded that itraconazole showed efficacy in reducing the size of aspergilloma and that this drug is useful also in terms of safety.

[Muramyl dipeptide derivative and its clinical application].

Muramyl dipeptide (MDP) and its synthetic derivatives which comes from the major constituent of bacterial cell wall has various biological activities as host defence mechanisms. One of the synthetic MDP derivatives, MDP-Lys (L 18), muroctasin has potent biological activities with less adverse reactions among various MDP derivatives. Muroctasin has proved to be safe to use clinically as the results of phase I clinical study. We attempted to evaluate its clinical usefulness and safety from the view point of restorative activity in leukopenia that was induced by cancer chemotherapy in patients with malignancies. It is concluded that muroctasin is effective as well as useful on the restorative activities against leukopenia in lung cancer patients after cancer chemotherapy, with the optimal daily dosage of 200 micrograms for six times by subcutaneous injections through phase II and phase III clinical cooperative studies in Japan. Supposed mode of action of muroctasin for granulocytosis may be the results of CSF production due to stimulation of macrophage by muroctasin. This first clinical success of restoration of leukopenia in patients with cancer receiving cancer chemotherapy by MDP derivative, muroctasin, might be not only advantageous for cancer chemotherapy and/or radiation therapy but also for preventing infections occurring in compromised host due to neutropenia in cancer patients by means of cytotoxic cancer chemotherapy or irradication.

[The studies on tuberculosis present research ideas for medicine].

I started my medical career as medical mycologist following the first case of deep-seated candidiasis in Japan. On the other hand I have been seeing many patients with tuberculosis as well as respiratory diseases. In my sense the seeing of mycobacterial infections may still remain lot of clinical, biological or immunological research ideas in future. I did the studies on muramyl dipeptide (MDP) which derived from cell wall substance of mycobacteria. MDP seemed to be one of the enhancing immunomodulator for host defense mechanism in particular in the immunocompromised host.

[A cooperative clinical study on the evaluation of an antibody detection test kit (MycoDot Test) for mycobacterial infections. Cooperative Study Group for MycoDot Test].

To determine the usefulness of a diagnostic kit for mycobacterial infection, we performed a five-hospital cooperative clinical study using serodiagnosis kits (MycoDot Test) to detect antibody for lipoarabinomannan (LAM) which is a membrane-derived component of mycobacterial species. We tested LAM antibody in the sera of patients with mycobacterial infection as well as healthy persons. Procedures for using the serodiagnosis kit are actually simple. Out of 130 cases of active pulmonary tuberculosis, 103 cases (79%) were positive for anti-LAM, and cases out of 24 cases of active atypical mycobacterial infection, 15 (63%) were positive. On the contrary, only 4% of healthy volunteers (1 out of 25 persons) were positive on this test. In conclusion, this diagnostic kit might be a useful test for early and supportive diagnosis of mycobacterial infections based on its sensitivity and specificity.

[A case of pulmonary tuberculosis complicated with gingival lesions].

A case of secondary gingival tuberculosis is presented. The case is 51 year-old male who had been suffering from undetected pulmonary tuberculosis visited a dentist because of chronic periodontal inflammation around the gingiva of the right upper and left lower molar teeth lasting for one year. The lesions remained unchanged and painful granulomatous swelling sustained in spite of the conservative treatment. The case was treated with the extraction of six teeth due to continued toothache. By pathohistological examination of gingiva and chest X-ray examination, the case was diagnosed as tuberculosis. Chest roentgenogram showed active pulmonary tuberculosis, and bacteriological examination of sputum showed tubercle bacilli. The administration of INH, RFP and EB was started, and the response to the treatment was good and the pain in the gingiva disappeared within three weeks. Secondary gingival tuberculosis is manifested as local granulomatous lesions with severe pain. The incidence of gingival tuberculosis is very rare, but we have to keep in mind that the oral tuberculosis secondary to pulmonary tuberculosis could occur.

[Comparison of the newly developed MB redox system with mycobacteria growth indicator tube (MGIT) and 2% Ogawa egg media for recovery of mycobacteria in clinical specimens].

The rate of recovery and the mean time to detection of mycobacteria in clinical specimens were determined in a newly-developed MB Redox system based on liquid medium, and the results were compared with those of MGIT and 2% Ogawa egg media. From 587 sputum specimens processed, totally 203 mycobacterial isolates were detected, of which 177 (87.2%) with MB Redox, 185 (91.1%) with MGIT and 133 (65.6%) with 2% Ogawa medium. The difference in the percentages of positive cultures between either of the two liquid media and 2% Ogawa medium was significant (p < 0.0001). The mean time to detection of the Mycobacterium tuberculosis complex was 17.5 days with MB Redox, 18.7 days with MGIT, and 26.2 days with 2% Ogawa medium. The contamination rates were 1.5, 1.7, and 4.1% for MB Redox, MGIT, and 2% Ogawa medium, respectively. In conclusion, both MB Redox and MGIT systems, based on liquid medium, are more efficient than 2% Ogawa medium for the recovery of mycobacteria in clinical specimens.

[A clinical phase II trial of ulinastatin (MR-20) for nephrotoxicity of cisplatin].

UNLABELLED: MR-20 was administered to 37 lung cancer patients who presented with nephrotoxicity due to cisplatin (CDDP) treatment in an 11-institution cooperative trial during the period from September 1990 through March 1992 to study its suppressive effect on the nephrotoxicity as well as its safety. The results are reported in this paper. METHODS: MR-20 was administered at 3 daily doses, ie. 150,000, 300,000 and 600,000 units, in a nonblind manner. An efficacy rate of 27.3% was achieved in the group at 150,000 units; 44.4% in the group at 300,000 units; and 45.5% in the group at 600,000 units. Serum Cr levels remained significantly lower in the group at 300,000 units during the treatment period than in the screening period. No adverse reactions to MR-20 were observed. According to the above results, it was anticipated that MR-20 would suppress the nephrotoxicity of CDDP treatment and be safe, and that the drug at a dose of 300,000 units daily would be useful in suppressing the nephrotoxicity.

[Antitumor effect of levamisole].

The inhibition of MH134 tumor growth which was transplanted into foot-pad of C3H mice was observed in levamisole treatment. The inhibitory mechanism of tumor growth by levamisole was studied by the transfer of spleen cells. It was suggested that activated macrophage within spleen cells by levamisole might be played a major role for inhibition.

[Mechanism of cancer metastasis and its inhibition--a point of view from clinical aspects].

Metastasis is one of the most distinct but complicated biological phenomena in cancer. It is hard to decide the most important prospect which must be performed at the present as well as in future studies on cancer metastasis. However, the final goal of the studies on metastasis can be simply concluded as that metastasis should be prevented before dissemination. In this review, we discussed several points to do research in metastasis.

[Induction of human monocyte-mediated tumor cell killing by alpha or beta interferon].

Human blood monocytes, separated on a continuous percoll gradient, were not cytotoxic to allogeneic A375 melanoma cells. The monocyte monolayers were activated to become tumoricidal by incubation for 24 hr with interferon (IFN)-alpha or beta at concentrations of more than 1,000 IU/ml. Significant and reproducible activation of the monocytes was achieved by incubating them with 10,000 IU/ml of IFN-alpha or IFN-beta for 24 hr. Similarly, suspended, but not plated, monocytes were activated to a tumoricidal state by interaction with IFN-alpha or IFN-beta. Monocytes that had lost tumoricidal activity during culture, were reactivated by a second exposure to IFN-alpha. Fluorescence analysis showed that the monocyte-rich adherent monolayers were contaminated with up 2.0% of natural killer (NK) cells. Pretreatment of isolated monocyte preparations with anti-NK cell monoclonal antibody (Leu-11b) to deplete them of NK cell activity did not inhibit the monocyte-mediated cytotoxicity. These results indicate that human monocytes are rendered tumoricidal by direct interaction with IFN-alpha or IFN-beta, although more than 1,000 IU/ml of IFN-alpha or IFN-beta is required for maximal expression of monocyte activation.

[A case of remission in diffuse pleural mesothelioma induced by anticancer chemotherapy with cisplatin and adriamycin].

A 56-year-old man was admitted to our hospital with complaints of dyspnea and right pleural effusion. Malignant pleural mesothelioma was diagnosed, based on the high level of hyaluronic acid in the pleural fluid and manifestation of malignant cells by pleural needle biopsy. Anticancer chemotherapy with cisplatin and intrapleural injections of adriamycin against this mesothelioma was carried out because of the presence of stage III by Butchart's classification. Regression of the tumor, decrease of pleural fluid and decrease of LDH level in effusions were observed after this chemotherapy. Cisplatin was easily transported into the pleural fluid, and its concentration was maintained for a satisfactorily long period.