Preclinical anticancer effects and toxicologic assessment of hepatic artery infusion of fine-powder cisplatin with lipiodol in vivo.

We conducted an in vivo study to evaluate the anticancer effect and toxicity of fine-powder cisplatin suspended in lipiodol (fCDDP/LPD suspension) after a single administration of three different doses to rats via the intrahepatic artery after transplantation of rat ascites hepatoma cells. The toxicity of the fCDDP/LPD suspension was also assessed in the same protocol in noncancer-bearing rats and the observed toxicologic changes were compared among groups administered saline (Sal), an aqueous solution of fCDDP (fCDDP/Sal solution), and LPD alone. In parallel with the toxicity test, plasma CDDP concentrations were compared between the fCDDP/LPD suspension and fCDDP/Sal solution. The mean weight of the tumors in the fCDDP/LPD suspension groups was significantly less than in the LPD-alone group. The pathologic changes in the liver observed in the fCDDP/LPD suspension group increased with dose, were more marked compared with those in the fCDDP/Sal solution and LPD-alone groups, and were reversible. No other toxicologic effects were observed. The concentration of CDDP in the plasma in the fCDDP/LPD suspension group was slightly lower than that in the fCDDP/Sal solution group. In conclusion, the results indicate that the fCDDP/LPD suspension has sufficient anticancer efficacy and tolerability for use in the clinical treatment of hepatocellular carcinoma.

[Tuberculosis of the elderly (above the age of 75) in national hospitals].

In Japan tuberculosis is becoming rapidly the disease of the elderly. We studied the background, the type and level of hospital cares needed, and the outcome of patients with pulmonary tuberculosis (sputum smear and/or culture positive) above the age of 75 who were admitted to 8 national hospitals during the period from January 1 to December 31, 1997. The study included 150 patients (male: 109, female: 41, mean age: 81.6), of whom 25% needed care in a single-bed room, 84.3% had underlying diseases (cardiovascular diseases: 43.3%, malignant diseases: 20.9%, neuro-psychiatric diseases: 13.4%), and 47.6% needed cares mainly in feeding and excretions. 92 patients (62.6%) improved and 45 patients (30.6%) died, of whom the cause of death was directly related to tuberculosis in 42. The mean hospital stay was 4.7 months. However, in 42 patients whose cause of death was related to tuberculosis, 66% died within 3 months, while in 102 patients who were discharged 71% stayed more than 3 months. The same comparison was done in 508 patients with bacteriologically proven tuberculosis above the age of 75 admitted to National Tokyo Hospital during the period from 1990 to 1999. The result was almost the same, among 133 patients died in the hospital 60% died within 3 months, while in 375 patients who were discharged hospital stay was more than 3 months in 70%. In the near future, the elderly will occupy more than 25% of the beds of the tuberculosis ward in Japan and most of them have underlying diseases other than tuberculosis. Because tuberculosis, once the disease of the young, is becoming rapidly the disease of the elderly, it is imperative for us to make necessary adjustments to meet this inevitable trend.

A study on the metabolism of etoposide and possible interactions with antitumor or supporting agents by human liver microsomes.

The metabolism of etoposide was investigated by using human liver microsomes and nine recombinant human cytochrome P450 (CYP) isoforms to identify the CYP isoform(s) involved in the major metabolic pathway (3'-demethylation) of etoposide as well as to evaluate the possible metabolic interactions with several antitumor or supporting agents. The 3'-demethylation of etoposide followed a Michaelis-Menten one-enzyme kinetic behavior in six human liver microsomal samples. The relationships were assessed with six different human liver microsomes between the 3'-demethylation of etoposide and metabolic activities for substrate probes of the respective CYP isoforms, showing a significant correlation (r = 0. 932, P < .01) only with 6beta-hydroxylation of testosterone, a marker substrate for CYP3A4. Inhibitor/substrate probes for CYP3A4, ketoconazole, troleandomycin, verapamil and cyclosporin, or supporting agents, vincristine and prednisolone, inhibited etoposide 3'-demethylation by human liver microsomes. p-Nitrophenol, a substrate for CYP2E1, also inhibited etoposide 3'-demethylation. Among the nine recombinant human CYP isoforms, CYP3A4 exhibited the highest catalytic activity with respect to etoposide 3'-demethylation, compared with the minor activities of CYP1A2 and 2E1. Collectively, these data suggest that etoposide 3'-demethylation is mediated mainly by CYP3A4 and to a minor extent by CYP1A2 and 2E1. Furthermore, some supporting agents (vincristine and prednisolone) and the substrates of CYP3A4, which may be coadministered with etoposide during the cancer chemotherapies, inhibit the etoposide 3'-demethylation activity in vitro. The results may provide clinical implications with respect to the possible metabolic interactions between etoposide and other drugs studied herein in patients with cancer undergoing etoposide concurrently with either of them.

[Evaluation of a new detection method of Mycobacterium tuberculosis in clinical specimens by amplification of ribosomal RNA and its clinical application].

A new rapid M. tuberculosis detection method, the Gen-Probe Amplified Mycobacterium Tuberculosis Direct Test (MTD), which allows direct detection of M.tuberculosis in clinical specimens by amplification of M.tuberculosis ribosomal RNA (rRNA), was evaluated. In the present study, MTD and conventional smear and culture examinations were performed on 225 sputum, 36 pleural fluid (PF), nine cerebrospinal fluid (CSF) and 41 bronchoalveolar lavage fluid (BALF) specimens. Among 225 sputum specimens from 132 patients, 20 culture-negative specimens from 14 patients and one specimen which was culture-positive for M.fortuitum on Ogawa's egg medium were MTD-positive. On the other hand, there were 6 M.tuberculosis culture-positive but MTD-negative specimens from 6 patients. The amount of bacilli in all six cases, however, was very few. The sensitivity of M.tuberculosis detection by MTD, as compared with Ogawa's egg medium and MB-Check, was 93 and 92%, respectively, in sputum specimens, and the specificity of MTD was 80 and 85%, respectively. Taking into account not only culture-positive specimens but also MTD-positive specimens from patients clinically diagnosed as active tuberculosis, the sensitivity and specificity of MTD were 93% and 99%, respectively. Among 36 PF specimens from 31 patients, of which 20 specimens from 16 patients were clinically diagnosed as tuberculous, only 4 from 4 patients were culture-positive for M.tuberculosis while 7 from 5 patients were MTD-positive. The results of MTD in 9 CSF specimens coincided well with those of MB-Check, but one MTD-positive specimen yielded a false-negative result with Ogawa's egg medium. Among 41 BALF specimens from 39 patients, only one was culture-positive while one culture-positive and 7 culture-negative specimens were positive on MTD. All these MTD-positive patients were later verified as having pulmonary tuberculosis either by transbronchial lung biopsy and other examinations, or by the clinical course of the disease, particularly the response to anti-tuberculosis drugs. Inconsistent results were seen in 21 sputum specimens from 15 patients, 4 PF specimens from 3 patients and 7 BALF specimens from 7 patients, all of which were culture-negative for M.tuberculosis but MTD-positive. None of these patients had any clinical findings inconsistent with diagnosis as tuberculosis. It was noted that MTD was very useful for rapid detection of M.tuberculosis in sputum as well as other clinical specimens. We observed the clinical course of 20 pulmonary tuberculosis patients by monthly examinations of sputum, using smears, Ogawa's egg medium, MB-Check and MTD. MTD-positivity rates fell down in parallel with decreased pulmonary tuberculous activity. During the clinical course, out of 61 MTD-negative specimens only 4 (6.6%) were culture-positive for M.tuberculosis, while out of 63 smear-and-culture-negative specimens 6 (9.5%) were MTD-positive. These data suggest that MTD is useful to assess the clinical activity and course of tuberculosis.

Cell-associated IL-8 in human blood monocytes: analysis by flow cytometry.

Several cell-associated cytokines, such as interleukin-1 (IL-1) and tumor necrosis factor, exist on the cell surface and are biologically active. Although extracellular IL-8, a potent chemotactic factor for primarily neutrophils, has been studied extensively, cell-associated IL-8 has barely been studied. In this study, we analyzed the intracellular and cell-surface IL-8 in human blood monocytes in vitro by using flow cytometry and predicted the biological activity of the cell-associated IL-8 in vivo. After fixation with paraformaldehyde, mononuclear cells were divided into two subgroups. One subgroup was left untreated to study cell-associated antigens, and the other subgroup was permeabilized with saponin to detect intracellular antigens. In lipopolysaccharide (LPS)-stimulated monocytes, IL-8 was detected solely intracellularly, whereas both the intracellular and cell-surface IL-1 beta was detectable. In a time-course study, the intracellular IL-8 increased in response to LPS stimulation, but the cell-surface IL-8 was undetectable throughout the course. In an LPS-stimulated monocytic cell line, both ELISA and flow cytometry detected the quantitative change of the intracellular IL-8. The dissimilar localization between IL-8 and IL-1 beta within cells was confirmed by the immunohistochemical analysis. In summary, LPS stimulation induced a time-dependent increase in intracellular but not cell-surface IL-8 in monocytes. Thus, it is unlikely that the cell-associated IL-8 is functioning physiologically. The semiquantitative flow cytometric procedure may be useful for simultaneous examination for cell-surface and intracellular cytokines.

[A case of AIDS with bronchial tuberculosis].

A case is 48 years-old Japanese man who had a history of frequent sexual contact with prostitutes in Thailand and the Philippines. He presented with chief complaint of chest discomfort in April 1995. His chest X-ray film showed right mediastinal lymph node swelling in other hospital and the sputum smear was strongly positive for acid fast bacilli. In May 1995, he was admitted to our hospital and serological tests for HIV were positive both by EIA and Western blot methods. The CD4 lymphocyte count was 167/microliters. He was diagnosed as a case of AIDS according to the criteria proposed by the AIDS surveillance committee of the Japanese Ministry of Health and Welfare. Although numerous tubercule bacilli were detected in sputum, the chest X-ray did not show abnormal shadow in lung fields. So the diagnosis of bronchial tuberculosis was suspected by these apparently contradictory findings and the bronchoscopy was performed. Biopsy specimen of the bronchial mucous membrane obtained by bronchoscopy confirmed the presence of acid fast bacilli by Ziehl-Neelsen's staining method, however, histological findings were atypical of tuberculosis. A month after the initiation of treatment with isoniazid, rifampicin and ethambutol and AZT, his clinical symptoms improved and the sputum smear and the culture tests for tubercule bacilli converted to negative. Complications of AIDS, (Pneumocystis carinii infection, Cytomegalo virus infection, Kaposi's sarcoma, etc) other than tuberculosis have not developed to date. In the past reports, we could not find reports of bronchial tuberculosis with AIDS. Tuberculous granuloma formation was scarce in this case, and it was suspected that bronchial tuberculosis with AIDS would show characteristic sign as same as pulmonary tuberculosis with AIDS.

[Blood gas changes in Duchenne type muscular dystrophy].

We studied changes in blood gas data from 32 patients with Duchenne type muscular dystrophy (DMD), who were followed for more than 5 years. Regression lines for each blood gas variable were obtained by the least square regression method, from 276 observations. The mean change per year was calculated by averaging the individual slopes. All blood gas variables (PaO2, PaCO2, pH, and [HCO3-]) correlated significantly with age the regression lines were: PaO2 = 95.6-0.45x Age, PaCO2 = 34.7 + 0.70x Age, pH = 7.397-0.0023x Age, [HCO3-] = 20.7 + 0.28x Age. The rates of change were -0.74 Torr/year for PaO2, 1.07 Torr/year for PaCO2, -0.0028 per year for pH and 0.55 mEq/l/year for [HCO3-]. The slope of each regression line was within the average individual yearly change +/- one standard deviation. AaDO2 did not change significantly with age, but chronic respiratory acidosis worsened because of alveolar hypoventilation. As an index of the severity of DMD, PaCO2 was better than PaO2, because the former was more strongly correlated with age and the values of PaCO2 deviated less from the the regression line.

Long-term course of bronchiectasis and bronchiolitis obliterans as late complication of smoke inhalation.

We describe the long-term course of a patient with bronchiectasis and bronchiolitis obliterans, both of which developed as late complications of a smoke inhalation injury. Sequential chest X-rays obtained during the observation period showed gradual progression of bronchiectasis from the saccular to the cystic type. Symptoms, spirometry and blood gas analysis, however, remained stable for 15 years. We believe that symptoms and physiological derangement were due mainly to bronchiolitis obliterans, and that once the pathophysiological condition had been established following the initial injury, it could be maintained by conservative medical management.

[Gastric metastasis of primary lung cancer].

A 74-year-old man was admitted to Keio University Hospital because of an abnormal shadow in the right upper lung field on chest roentgenogram. Bronchoscopic examination revealed malignant cells, by which large cell carcinoma of the lung was diagnosed. Systemic survey for metastasis disclosed an ulcerated tumor on the greater curvature of the stomach. Endoscopic examination showed an elevated submucosal tumor suggesting submucosal metastasis of lung cancer to the stomach. A review of 473 autopsies of primary lung cancer at Keio University from 1958 to 1987 showed 16 cases with gastric metastasis (3.4%). Two of 16 cases were detected clinically. Gastric metastases were proven in 15.6% of autopsy cases of large cell carcinoma of the lung. Forty-three cases of primary lung cancer with gastric metastasis were reported in the Japanese literature from 1965 to 1990. Thirteen of these cases had large cell carcinoma.

[A case of congenital antithrombin III deficiency with pulmonary thromboembolism and cerebellar infarction].

A 41-year old woman was admitted to a local hospital because of leg pain, chest pain and dyspnea after taking estrogen for two months for irregular menstruation. On admission to the hospital, her lung scintigram showed multiple segmental perfusion defects in the right lung. Pulmonary angiography showed several thrombi in the proximal pulmonary artery. A diagnosis of pulmonary thromboembolism was made. AT-III activity was found to be only 50% of normal value, indicating that the pulmonary thromboemboli were due to AT-III deficiency. The patient was then put on anticoagulants for two years. When she was 43-year-old, she was admitted to Keio University Hospital because of worsening of dyspnea. After admission her dyspnea had got better due to bed rest. Her brain CT, which was performed because of her gait disturbance, indicated past right cerebellar infarction. We gave her 2 mg of warfarin and maintained a thrombotest of about 30%. The investigation of her family revealed that her son also showed decrease AT-III activity, indicating a congenital abnormality. 26 families with this disease have been reported in Japan. Most had venous thromboemboli, but only a few cases had brain infarction as well. In addition, it is suggested that an estrogen therapy for irregular menstruation may have contributed, in this particular case, to the onset of thromboemboli.

Detection of tracheal stenosis by frequency analysis of tracheal sounds.

To develop a simple noninvasive method for detecting tracheal stenosis, tracheal sounds were analyzed using fast-Fourier transform. The subjects were all female and included 5 normal volunteers and 13 patients with tracheal stenosis mostly secondary to thyroid cancer (11 extrathoracic and 2 intrathoracic lesions). Tracheal sounds were recorded during spontaneous breathing and were digitized with an analog-to-digital converter. Pulmonary functions, including forced expiratory volume in 1 s (FEV1) expressed as percentage of vital capacity, peak expiratory flow rate (PEFR), the ratio of FEV1 to PEFR (Empey's index), and the ratio of expiratory to inspiratory flow rates at 50% vital capacity, were measured. A computed tomography scan was used to obtain the tracheal minimum cross-sectional area. Whereas PEFR demonstrated a weak correlation with the stenotic area, FEV1%, Empey's index, and the ratio of expiratory to inspiratory flow rates at 50% vital capacity did not. The power of the fast-Fourier transform spectrum of normal tracheal sounds decreased as the frequency increased up to 500 Hz. A small spectral peak was observed at approximately 1 kHz. Patients with significant tracheal stenosis demonstrated an increase in the peak spectral power at approximately 1 kHz and in the mean spectral power from 600 to 1,300 Hz in their tracheal sounds. In patients with extrathoracic lesions, the peak and mean spectral powers correlated well with the area of the stenosis as defined by computed tomography scan. In patients with intrathoracic lesions, abnormalities in the pulmonary functions as well as tracheal sound spectra appeared more evident despite milder stenoses.(ABSTRACT TRUNCATED AT 250 WORDS)

[Effects of antioxidant enzymes in erythrocytes on pulmonary arterial pressure during hypoxic gas breathing].

Using isolated rabbit lungs (n = 72) perfused at a constant flow of 70 ml/min, we analyzed whether the antioxidant system in erythrocytes significantly contributes to maintaining pulmonary vascular responsiveness to alveolar hypoxia (HPV) in normal lungs. As a measure of HPV, we observed the difference (delta P) between mean pulmonary arterial pressure during ventilation with normoxic gas mixture (21% O2, 5% CO2 in N2) and that during hypoxic gas breathing (3% O2, 5% CO2 in N2). Autologous erythrocytes obtained from the animals treated with various substances inhibiting either superoxide dismutase (SOD), the anion channel of the membrane, catalase (CAT) or glutathione peroxidase (GSH-Px). Subsequently, delta P was systematically measured in the perfusate, whose hematocrit was adjusted to 6-7% with normal or treated erythrocytes as described above. Further, the effects of adding SOD (75 U/ml) or CAT (1000 U/ml) to the perfusate on delta P were examined. The following results were obtained. (1) Inhibition of the superoxide scavenging mechanism in erythrocytes (SOD and anion channel) exerted no significant influence on delta P. (2) Inhibition of hydrogen peroxide scavengers in erythrocytes did not alter the scope of delta P. (3) Addition of either SOD or CAT to the perfusate did not show any significant effect on delta P. The findings are highly consistent with the idea that HPV in normal lungs is essentially independent of antioxidant enzymes in erythrocytes, which are expected to be one of the important factors determining the total capacity to deal with oxidant stress in the lung.

[Effect of diffusion limitation on gas exchange in the lung--analysis in oleic acid-induced lung injury].

To assess the effect of diffusion limitation on gas exchange in injured lungs with non-cardiogenic pulmonary edema, an experimental model of acute lung injury with alveolar flooding was produced in six mongrel dogs by intravenously injecting oleic acid at 0.06 ml/kg. The effect of diffusion limitation was quantitatively examined by measuring the excretion (E) of three indicator gases (acetylene, ethylene and freon-22) with differing solubility (lambda) and tissue diffusivity (d). The indicator gases were dissolved in normal saline and infused at a constant rate through a peripheral vein. Since acetylene and ethylene have nearly identical of d but differing lambda, the difference in E values of these two gases may solely reflect the effect of uneven distribution of ventilation-perfusion ratios (VA/Q) in the lung. Thus, measured E values of acetylene and ethylene allowed us to approximately predict the E of freon-22, the value corresponding to the condition where d of freon-22 was taken to be equal to that of acetylene or ethylene. The difference between predicted and measured E values of freon-22 is indicative of the limitation of diffusion in the lung periphery. In all the lungs studied, measured E values of freon-22 were consistently smaller than those predicted from acetylene and ethylene, leading to the conclusion that gas exchange in injured lungs with widespread pulmonary edema was partly impaired by diffusion in aqueous media.

[Assessment of pulmonary aerosol deposition and epithelial permeability in 99mTc-DTPA inhalation scintigram].

The degree of lung injury in bronchiolo-alveolar lesions may be quantitated from the pulmonary epithelial permeability estimated by 99mTc-DTPA (diethylene triamine penta acetate) aerosol inhalation scintigram. However, significant aerosol deposition sometimes occurs in the central airways and obscures the permeability change in the lung periphery. The radioaerosol deposition pattern and its effect on assessing the pulmonary epithelial permeability was studied. 99mTc-DTPA aerosol scintigraphy was performed in 47 patients with pulmonary fibrosis (PF), 12 patients with chronic obstructive pulmonary diseases (COPD), and 27 non-smoking and 17 smoking healthy volunteers. The scintigraphic images of the lungs were classified into 4 grades, 0; homogeneous distribution, 1; patchy distribution, 2; hot spots with partial defect, and 3; hot spots with little deposition in the lung field. The rate constant was used as a parameter for the permeability. The smokers and patients with PF showed increased kep values of 2.36 +/- 1.21%/min (mean +/- SD) and 2.49 +/- 1.29%/min as compared with the nonsmokers with 0.94 +/- 0.27%/min, respectively. The nonsmokers, smokers and 36 patients with PF were classified as deposition grade 0 or 1, suggesting good aerosol penetration to the lung periphery. All patients with COPD showed either grade 2 or 3 deposition. Aerosol deposition in the central airways can cause underestimation of the permeability because of the thicker lining layer in the bronchus than in the alveolus. In conclusion, the aerosol deposition pattern should be analyzed when the method is applied clinically to assess the permeability of the bronchiolo-alveolar epithelium.

[Detection of radiation-induced lung injury by 99mTc-DTPA aerosol inhalation method].

We prospectively monitored pulmonary 99mTc-DTPA (diethylene triamine penta acetate) clearance in patients who received chest radiation therapy, in order to determine whether this method allows us to predict the development of radiation pneumonitis. The rate constant of pulmonary 99mTc-DTPA clearance (k; %/min) was used to assess pulmonary epithelial damage. Fifteen nonsmoking patients who underwent radiation therapy were studied. The subjects included 4 patients who had already developed radiation pneumonitis at the time of study, and 11 in whom we prospectively observed 99mTc-DTPA clearance serially during the course of chest radiation therapy. In the 4 patients with pre-existing radiation pneumonitis, the mean k value obtained from the area with infiltration on the chest X-ray was significantly greater than that from the opposite lung (p < 0.02). In the prospective study, 3 out of 11 patients developed radiation pneumonitis. The mean k of the irradiated lung field in the 8 patients who did not develop radiation pneumonitis was unchanged. The mean k value obtained in the 3 patients who did develop radiation pneumonitis increased just before the onset, and further increased when the disease manifested clinically. We conclude that pulmonary 99mTc-DTPA clearance may be useful for predicting the development of radiation pneumonitis.

[A case of slowly-growing localized malignant mesothelioma].

We report a case of localized malignant mesothelioma. The patient was a 37-year-old male, admitted to our hospital because of a solitary nodular shadow in the right middle lung field noted on chest X-ray. Five years prior to admission, a solitary nodular shadow had been seen in the same area on chest X-ray, and the diameter of the tumor had gradually increased. A transbronchial biopsy specimen revealed proliferation of spindle-shaped tumor cells, suggesting fibrosarcoma. No other lesions were identified as primary foci, so we made the tentative diagnosis of primary pulmonary sarcoma prior performing right upper lobectomy. The resected specimen revealed that the tumor, which partially touched the pleura, contained a number of large and small cystic spaces, and was composed of numerous spindle-shaped tumor cells. Some of the tumor cells were immunohistochemically positive for cytokeratin and epithelial membrane antigen, and many mitotic figures were noted. Thus, we made the histopathological diagnosis of localized malignant mesothelioma.