Frequent epigenetic inactivation of Wnt antagonist genes in breast cancer.

Although mutation of APC or CTNNB1 (beta-catenin) is rare in breast cancer, activation of Wnt signalling is nonetheless thought to play an important role in breast tumorigenesis, and epigenetic silencing of Wnt antagonist genes, including the secreted frizzled-related protein (SFRP) and Dickkopf (DKK) families, has been observed in various tumours. In breast cancer, frequent methylation and silencing of SFRP1 was recently documented; however, altered expression of other Wnt antagonist genes is largely unknown. In the present study, we found frequent methylation of SFRP family genes in breast cancer cell lines (SFRP1, 7 out of 11, 64%; SFRP2, 11 out of 11, 100%; SFRP5, 10 out of 11, 91%) and primary breast tumours (SFRP1, 31 out of 78, 40%; SFRP2, 60 out of 78, 77%; SFRP5, 55 out of 78, 71%). We also observed methylation of DKK1, although less frequently, in cell lines (3 out of 11, 27%) and primary tumours (15 out of 78, 19%). Breast cancer cell lines express various Wnt ligands, and overexpression of SFRPs inhibited cancer cell growth. In addition, overexpression of a beta-catenin mutant and depletion of SFRP1 using small interfering RNA synergistically upregulated transcriptional activity of T-cell factor/lymphocyte enhancer factor. Our results confirm the frequent methylation and silencing of Wnt antagonist genes in breast cancer, and suggest that their loss of function contributes to activation of Wnt signalling in breast carcinogenesis.

Efficacy and outcomes of clarithromycin treatment for pulmonary MAC disease.

SETTING: Retrospective review of patients with pulmonary Mycobacterium avium complex (MAC) disease treated with clarithromycin. OBJECTIVES: To investigate whether the short-term response to treatment predicts long-term outcomes, and to analyse what explanatory variables are associated with the efficacy and outcome of treatment. DESIGN: Sputum conversion rates in short- and long-term outcomes were evaluated for 111 patients. Respectively 9 and 10 explanatory variables were analysed for their association with both response and outcomes. RESULTS: Eighty-four patients (75.7%) showed good short-term response and 94 (82.0%) showed good long-term outcomes. Women and patients with satisfactory nutrition status showed good short-term response. Patients with small lesions and those treated for >12 months after sputum conversion showed good long-term outcomes. Patients who showed good short-term response, in the group with large lesions, showed significantly good long-term outcomes (P = 0.0382). CONCLUSION: There were differences between prognostic factors reflecting short-term response and long-term outcomes. The short-term response predicts long-term outcomes in certain groups divided by prognostic factor. To establish standard treatment for pulmonary MAC disease, it is important to determine a standardised method of evaluation of treatment taking such factors into consideration.

Molecular epidemiology of tuberculosis in the north Hokkaido district of Japan.

SETTING: The incidence of tuberculosis in Japan has fallen rapidly in the past 50 years, to 27.9 cases per 100,000 population in 2001. OBJECTIVE: To assess the molecular epidemiology of tuberculosis in the north Hokkaido district, Japan. DESIGN: Restriction fragment length polymorphism analysis was consecutively performed on Mycobacterium tuberculosis strains from patients with pulmonary tuberculosis from April 1999 to March 2002. RESULTS: Of 229 patients, strains from 227 (99.1%) were available for analysis. The mean age of the patients analysed was 69.4 years. There was one immigrant patient. Two hundred and seven patients had six or more copies of IS6110. Of these, 16 (7.7%) in eight clusters had identical patterns, 120 (58.0%) belonged to five groups that had similar patterns (Dice coefficient >0.7) and 80 (38.6%) belonged to the 35 groups with the most common patterns (Dice coefficient >0.9). CONCLUSION: These results may reflect the epidemiological characteristics, age and migration of the residents and the incidence of tuberculosis of the area, and also those of Japan: many elderly patients were infected in the past, when the incidence of tuberculosis was very high.

Impaired interleukin-1beta converting enzyme (ICE) activity in patients with pulmonary tuberculosis.

We measured the activity of IL-1beta converting enzyme (ICE) of monocytes from pulmonary tuberculosis (PTB) patients. Peripheral blood mononuclear cells (PBMCs) and serum were collected from PTB patients (n = 16) and healthy subjects (n = 6). Serum IL-18 levels were significantly higher in PTB patients (515.9 +/- 326.5 pg/ml) than in controls (108.7 +/- 65.2 pg/ml) (P < 0.01), but gradually decreased with tuberculosis therapy. ICE activity was significantly lower in TB patients (65.3 +/- 34.4 vs. 142.2 +/- 75.6 U/mg) (P < 0.05). Serum IL-18 levels inversely correlated with ICE activity (P < 0.05). Altered ICE activity may explain why there is reduced IFN-gamma production from PBMCs; however, serum IL-18 is probably not regulated by ICE in monocytes in TB patients.

Increased vascular endothelial growth factor in acute eosinophilic pneumonia.

Acute eosinophilic pneumonia (AEP) is associated with the presence of diffuse pulmonary infiltrates on the chest radiograph and an increased number of eosinophils and an elevation of interleukin (IL)-5 levels in bronchoalveolar lavage (BAL) fluid. Vascular endothelial growth factor (VEGF) is a constitutively expressed protein encoded by messenger ribonucleic acid in human eosinophils and is released following stimulation with IL-5. However, the roles of IL-5 and VEGF in the pathogenesis or activity of this disease have not been clarified. The authors investigated the cells and the levels of these two factors in BAL fluid in five AEP patients and five normal controls before and after corticosteroid treatment. The absolute number of eosinophils-mL(-1), IL-5 and VEGF levels in patients before treatment were higher than in controls (53.8 versus 0.3 x 10(4) x mL(-1), 490.1 versus 5.2 pg x mL(-1) and 643.0 versus 133.9 pg x mL(-1), respectively). IL-5 and VEGF rapidly decreased to the control level in parallel with clinical improvement. The relationship between eosinophilia and IL-5 and VEGF levels was strongly significant. Elevated interleukin-5 in the lung may initiate the recruitment of eosinophils and enhance the release of mediators, such as vascular endothelial growth factor from eosinophils, which, in turn, increases the permeability of blood vessels.

Disseminated Mycobacterium avium complex (DMAC) in an immunocompetent adult.

A female patient with multiple osteomyelitis and pulmonary Mycobacterium avium disease visited an orthopaedic clinic with back pain. Systemic bone scan showed multiple sites of increased radioactivity in the vertebral bodies, right scapula, femurs and ribs. M. avium was isolated from sputum and a sample aspirated from the right scapula. The route of infection was unknown as there was no history of trauma or surgery. HIV testing was negative. As there was no underlying immunological disease she was diagnosed as disseminated M. avium complex (DMAC) disease in an immunocompetent adult. Cytokine production on several stimuli from peripheral blood mononuclear cells was similar to that in pulmonary M. avium patients. Sequence analysis of IFN-gamma receptor revealed no nucleotide substitution. We detected serotypes 1, 2 and 4 from mycobacteria cultured from the right scapula, and conclude that this case could be the result of undetected immune deficiency and/or unrecognised virulence of the infecting isolate.

[Two cases of acute eosinophilic pneumonia--analyses of cytokines in BALF before and after treatment].

Acute eosinophilic pneumonia (AEP) has been well recognized as sudden onset respiratory failure characterized by an increased eosinophilic cell count in the bronchoalveolar lavage fluid (BALF). Although several lines of research have demonstrated that the Th2 cytokine network is likely to play a pivotal role in the development of AEP, that mechanism has not been clearly understood yet. Previous reports published in Japan demonstrated that the BALF CD4/CD8 ratio in patients with AEP ranged between 1.1 and 5.8. In this report, we describe 2 cases of AEP in which the CD4/CD8 ratios were 0.3 and 7.8 when present. We measured IL-4, IL-5, IL-10 and IFN gamma in the BALF before and after treatment in these patients. IL-5 decreased drastically after the treatment phase, while the other cytokines did not change much. We concluded that the IL-5 concentration in the BALF is probably related to the development of AEP. In addition, we also speculated that the CD4/CD8 ratio in BALF from patients with AEP may be affected whether a patient has an atopic background or not.

Plasma von Willebrand factor activities in vascular Parkinsonism: comparison with Parkinson's disease.

We measured the level of plasma von Willebrand factor (vWF), a marker of vascular endothelial damage, in patients with vascular parkinsonism (VP) to clarify whether the level of plasma vWF is useful in the diagnosis of VP. We examined 26 patients with VP, 26 with Parkinson's disease (PD), 26 with cerebral infarct (CI), and 26 age-matched normal controls (NC). The vWF was measured by the ristocetin cofactor assay. The level of plasma vWF in the VP group was significantly higher than that in the PD or in the NC groups (P < .01). However, there was no difference between the VP group and the CI group. These findings suggest marked endothelial cell damage in patients suffering from VP and CI. Thus, the level of plasma vWF may be useful in diagnosing VP.

An adult case of mumps brainstem encephalitis.

We present an adult case of mumps brainstem encephalitis. He was successfully treated with steroid pulse therapy and recovered completely except for persistent dysuria. He had not been vaccinated and had no history of acute mumps infection. We consider that encephalitis in this case was caused by a reversible autoimmune process triggered by mumps infection. We emphasize the usefulness of pulse therapy for the treatment of some cases of mumps brainstem encephalitis in addition to the importance of mumps vaccination to prevent such a severe complication as encephalitis.

[Pulmonary infection caused by Mycobacterium gordonae in immunocompetent patient].

We report a case of 67-year-old female immunocompetent patient admitted to our hospital because of hemoptysis. Computed tomography (CT) of the lung showed bronchiectasis in the right S5 and small nodules in the right S6 and left S5. The cultures of sputum and bronchial washing specimen repeatedly revealed acid-fast bacilli identified as Mycobacterium gordonae (M. gordonae) by DNA-DNA hybridization (DDH) method. Thus, she was diagnosed to be infected with M. gordonae. She was treated with isoniazid, rifampicin, ethambutol and streptomycin. After treatment, the cultures of sputum and bronchial washing specimen converted to negative, and the chest CT showed improvement of small nodules. M. gordonae is a nontuberculous mycobacterium of very low pathogenic potency. Recently there have been a few reports of infection by M. gordonae not only in immunocompromised patients but also in immunocompetent patients. These cases were considered to be sensitive to initial standard antimycobacterial therapy, therefore, it is important to examine for M. gordonae in cases suspected of nontuberculous mycobacterial infection.

[Two cases of pulmonary disease due to Mycobacterium szulgai].

This paper describes with two patients with pulmonary disease due to Mycobacterium szulgai. The first patient was a 67-year-old man who consulted a doctor at the outpatient clinic of the Internal Medicine of our hospital, complaining with hemosputum. A chest X-ray showed an infiltrative shadow in the right upper lobe. A smear test of the sputum was negative but a culture was positive for mycobacteria. Second patient was a 37-year-old man who was admitted to our hospital, complaining with cough and fever. A chest X-ray showed an infiltrative shadow with cavity in the right upper lobe. A smear test was positive and culture was positive for mycobacteria. Cultured isolates of the two cases were indentified as M. szulgai. These two patients were treated with isoniazid, rifampicin and ethambutol daily. Their clinical symptoms improved and their sputum smears and cultures converted to negative for mycobacteria.

[Changes in atrial natriuretic peptide and pulmonary hemodynamics induced by exercise in patients with chronic pulmonary emphysema].

We studied the changes in pulmonary hemodynamics induced by arm-stretching exercise in 12 patients with chronic pulmonary emphysema (CPE) and 9 control subjects (Controls). Both patients and control subjects underwent right heart catheterization with inspired gas analysis at rest and during exercise for the 6 minute-exercise test. Mean pulmonary arterial pressure (mPAP) in patients with CPE significantly increased from 18.5 +/- 3.9 mmHg at rest to 25.7 +/- 5.1 mmHg during exercise without decreased oxygen tension of the arterial blood. Both mPAP and total pulmonary resistance increased in proportion to the increases in cardiac output. Mixed venous plasma atrial natriuretic peptide (ANP) was significantly evaluated during exercise in patients with CPE, but did not in Controls. There were a significantly positive relationships between ANP and mPAP, and a significantly negative relationships between ANP and PvO2 during exercise. These results suggest that pulmonary hypertension during light exercise in cases of CPE may be caused by deterioration of the pulmonary capillary bed, and that ANP may be a useful indicator for evaluating pulmonary hypertension in patients with CPE.

[Effect of bisoprolol, a beta 1-selective beta-blocker, on lipid and glucose metabolism and quality of life in elderly patients with essential hypertension].

The present study investigated the effect of bisoprolol, a beta 1-selective beta-blocker without intrinsic sympathomimetic activity (ISA), on lipid and glucose metabolism and quality of life (QOL) in elderly patients with essential hypertention. Bisoprolol at doses of 5-10 mg was administered once daily for 12 weeks to 60 non-elderly and 21 elderly outpatients with mild to moderate essential hypertension. In both groups bisoprolol significantly decreased both systolic and diastolic blood pressures and significantly reduced pulse rates to the same extent. The levels of serum cholesterol, HDL-cholesterol and triglyceride, and the response of plasma glucose and insulin to 75 g oral glucose load, were not changed in either group by the bisoprolol treatment. Bisoprolol significantly improved QOL in both groups. Bradycardia, a side effect attributable to bisoprolol, was noted in only one patient in the elderly group. These results suggest that bisoprolol is a safe and useful antihypertensive drug in elderly and non-elderly patients with essential hypertension.

[Effects of pimobendan on pulmonary hypertension in patients with chronic pulmonary emphysema].

Pimobendan is a positive inotropic agent that dilates blood vessels and sensitizes the myocardium to calcium. We studied the clinical usefulness of single oral doses of pimobendan (2.5 mg) in 8 patients with pulmonary hypertension caused by chronic pulmonary emphysema. We measured the short-term effects of pimobendan on pulmonary hemodynamics and gas exchange. Pimobendan significantly decreased mean pulmonary artery pressure from 23.4 +/- 3.4 to 18.5 +/- 2.5 mmHg, and total pulmonary resistance from 383 +/- 111 to 281 +/- 81 dyne.sec.cm-5, while oxygen delivery and cardiac output increased significantly (oxygen delivery: from 81.8 +/- 10.2 to 93.5 +/- 16.2 ml O2/min. cardiac output: from 4.98 +/- 0.76 to 5.77 +/- 1.49 l/min, means +/- SD). Systemic vascular resistance was not significantly reduced. The ratio of pulmonary to systemic vascular resistance was reduced, but not significantly. Arterial oxygen tension (PaO2) and arterial carbon dioxide tension (PaCO2) did not change significantly. These results indicate that pimobendan may be useful in the treatment of patients with secondary pulmonary hypertension.

Pentoxifylline potentiates the antitumor effect of cisplatin and etoposide on human lung cancer cell lines.

The effect of pentoxifylline (PENT) on the sensitivity of two human lung adenocarcinoma cell lines, PC-9 and PC-14, to cisplatin (CDDP) and etoposide (ET) was studied. PENT at 0.5 and 1 mM enhanced the cytotoxicity of CDDP and ET on PC-14 and PC-9, respectively. Isobologram analyses of IC50 data, as well as combination index calculations, revealed that PENT had an additive or a synergistic effect when applied in combination with CDDP or ET, respectively. PENT potentiated the antitumor effect of ET in a nude-mouse xenograft model using PC-14 cells, when PENT was administered at 150 mg/kg subcutaneously for 6 days. Flow cytometry revealed that PENT decreased the accumulation of cells in the G2+M phase caused by CDDP when using PC-9 cells. However, PENT did not remarkably alter the accumulation of cells in G2+M caused by ET. These results suggest that PENT enhanced the antitumor effects of CDDP additively and those of ET synergistically. The enhancement mechanism probably differs between CDDP and ET. PENT needs more study to elucidate its potency as a new agent for combination chemotherapy.

[A case of chronic tuberculous pyothorax associated malignant lymphoma].

The patient was 69-year-old male. He has a history of treatment for tuberculosis by artificial pneumothorax about 47 years ago. He was admitted an another hospital under the diagnosis of tuberculous pyothorax. He was transferred to our hospital because of chest pain and fever. Laboratory findings on the admission were as follows: ESR was 120 mm/hr, CRP was 20.22 mg/dl and other data were almost within normal limits. Chest X-ray showed a massive shadow in the right lower lung field, adjacent to the chest wall. Computed tomography (CT) showed tumor shadow with low density and invasions into the adjacent chest wall. Histological examination of surgically excised tumor biopsy revealed malignant lymphoma. The patient's condition improved and the size of tumor decreased temporarily by chemotherapy. Then, he began to complain of chest pain and high fever, and tumor in the chest wall invaded into the whole chest wall. He died of disseminated intravascular coagulation despite continuing chemotherapy. Postmortem examination revealed the following findings : the tumor existed mainly in the parietal pleura or the chest wall, adjacent to the lesion of pyothorax, and immunohistochemical examination showed that tumor was malignant lymphoma, diffuse, large B-cell type. Recent studies have shown a close association between EBV infection and pyothorax-associated lymphoma. We have to keep in mind the possible development of malignant lymphoma following tuberculous pyothorax, when we see patients complaining of fever or chest pain with tuberculous pyothorax.

[A case of humidifier lung associated with BHL].

We encountered a rare case of hypersensitivity pneumonitis associated with bilateral hilar lymphadenopathy (BHL). The patient was a 53-year-old male, who developed dry cough and shortness of breath when using a humidifier since 1982. He was admitted to our hospital for further evaluation in 1987. Chest X-ray films showed BHL and ground glass appearance in the bilateral lung fields. Pulmonary function test indicated disturbance of diffusing capacity. Transbronchial lung biopsy revealed interstitial pneumonitis, and lymph node biopsy by mediastinoscopy showed lymphoid sinus histiocytosis without noncaseating granuloma. Provocation test using the humidifier was positive, and the diagnosis of humidifier lung with BHL was made.

[Randomized trial of cisplatin plus ifosfamide versus cisplatin plus vindesine for non-small cell lung cancer (NSCLC)].

Between 2/87 and 10/89, 83 patients (pts) with previously untreated NSCLC were assigned at random to receive either the PI regimen (cisplatin 80 mg/m2 dl and ifosfamide 2 g/m2 d 1, 2, 3, q 3-4 wks) or the PV regimen (cisplatin 80 mg/m2 d 1 and vindesine 3 mg/m2 d 1, 8, 15, q 3-4 wks). Three pts (2 PI regimen, 1 PV regimen) were ineligible for the study and 13 pts (7 PI regimen, 6 PV regimen) did not complete it. Thirty-three PI regimen pts and 34 PV regimen pts completed 2 or more cycles of the treatment and were evaluated. Patient characteristics were almost identical in both groups in terms of sex, age, pathological types, stage, performance status and number of chemotherapy cycles. For the PI regimen, there were 1 C.R. and 6 P.R., and the response rate was 21.2% (7/33). In the PV regimen, there were 11 P.R., and the response rate was 32.4% (11/34). The median survival time (MST) for PI regimen was 29 weeks (range 12-156 wks) and for PV regimen 40 weeks (range 8-138 wks). The difference in the survival curves for the 2 regimens was not statistically significant. Toxicities for the 2 regimens were similar except for greater leukopenia and one treatment-related death due to renal toxicity on PI regimen. These results suggest that PI regimen is not superior to PV regimen in the treatment of NSCLC.

A phase III comparison of etoposide/cisplatin with or without added ifosfamide in small-cell lung cancer.

A total of 92 patients with small-cell lung cancer (SCLC) were randomized to receive cisplatin (80 mg/m2, day 1)/etoposide (100 mg/m2, days 1, 3, 5) (PE) or cisplatin (80 mg/m2, day 1)/etoposide (100 mg/m2, days 1, 3, 5)/ifosfamide (2 g/m2, days 1, 2, 3) (PEI) combination chemotherapy. After 2 courses of chemotherapy, patients with limited disease (LD) received chest irradiation of 40-50 Gy. Of the 89 patients who could be wholly evaluated, the overall response rate was 78% for PE and 74% for PEI therapy (NS). For all patients the complete response (CR) rates were 14 versus 21%, respectively, and 22 versus 30% for LD. However, the median survival times for all patients were 55 weeks for PE therapy versus 56 weeks for PEI therapy (NS). The 2-year survival rates were 15 and 17%, respectively, for all patients (NS). There was no difference in the duration of response between PE and PEI therapy in cases with CR or partial response. However, severe leukopenia (< 2,000/mm3) occurred more often after PEI (73%) than after PE (44%) therapy (p < 0.05). These results suggest that PEI is not superior to PE chemotherapy in SCLC. The use of ifosfamide in multimodality treatment regimens needs to be studied further.