Thy28 protects against anti-CD3-mediated thymic cell death in vivo.

Apoptotic cell death plays a pivotal role in the development and/or maintenance of several tissues including thymus. Deregulated thymic cell death is associated with autoimmune diseases including experimental autoimmune encephalomyelitis (EAE), a prototype murine model for analysis of human multiple sclerosis. Because Thy28 expression is modulated during thymocyte development, we tested whether Thy28 affects induction of EAE as effectively as antigen-induced thymocyte deletion using Thy28 transgenic (TG) mice. Thy28 TG mice showed partial resistance to anti-CD3 monoclonal antibody (mAb)-induced thymic cell death in vivo, as assessed by annexin V-expression and loss of mitochondrial membrane potential. The resistance to anti-CD3 mAb-induced cell death in Thy28 TG mice appeared to correlate with a decreased c-Jun N-terminal kinase phosphorylation and reduced down-regulation of Bcl-xL. Moreover, thymic hyperplasia was detected in Thy28 TG mice, although thymocyte development was unaltered. Development of peripheral lymphoid tissues including spleen and lymph nodes was also unaltered. Thy28 TG spleen T cells showed an increased production of IFN-γ, but not IL-17, in response to both anti-CD3 and anti-CD28 mAbs. Finally, Thy28 TG mice displayed accelerated induction of EAE as assessed by disease incidence, clinical score, and pathology following immunization with myelin oligodendrocyte glycoprotein compared with control WT mice. These findings suggest that modulation of Thy28 expression plays a crucial role in the determination of thymic cell fate, which may contribute to the development of EAE through proinflammatory cytokine production.

Cytochrome c release, mitochondrial membrane depolarization, caspase-3 activation, and Bax-alpha cleavage during IFN-alpha-induced apoptosis in Daudi B lymphoma cells.

Interferon-alpha (IFN-alpha) displays antitumor action by inducing direct cytotoxicity against tumor cells in addition to generation of cytotoxic cells. The IFN-alpha-induced direct cytotoxicity is at least partly due to induction of apoptosis. In the present study, we examined signaling pathways implicated in IFN-alpha-induced apoptosis in Daudi cells. Release of cytochrome c from mitochondria to cytosol was found after 12 h incubation with IFN-alpha, followed by a decline in mitochondrial membrane potential (Delta psi(m)) and procaspase-3 activation at 24 and 36 h, respectively. Cleavage of endogenous Bax-alpha (21 kDa), generating an 18-kDa fragment (p18 Bax-alpha), was found at 36 h. Although the endogenous p21 Bax-alpha was located in both cytosol and mitochondrial membranes, the p18 Bax-alpha resided only on mitochondrial membranes. IFN-alpha-induced apoptosis occurred 48 h after stimulation, with a further increase in proportion up to 72 h. Pretreatment with pancaspase inhibitor Z-VAD-fmk substantially inhibited the IFN-alpha-mediated Bax-alpha cleavage and apoptosis, but not the decline in Delta psi(m), suggesting the possibility that caspase-3 activation is implicated in the Bax-alpha cleavage, probably leading to amplification of the apoptotic processes. Our results suggest that modulation of endogenous p21 Bax-alpha is implicated in IFN-alpha-induced apoptosis.

Participation of Bax-alpha in IFN-alpha-mediated apoptosis in Daudi B lymphoma cells.

Although interferon-alpha (IFN-alpha) has proved beneficial in the treatment of some tumors, the basis for this is still uncertain. In this study, we examined the effects of IFN-alpha on the growth of tumor cells in vitro, using the Daudi line of B lymphoma cells as a model. There was a dose-dependent accumulation of these cells in the G1 phase of the cell cycle 24-48 h from the time of exposure to IFN-alpha. This was followed between 48 h and 96 h by an increasing degree of apoptosis, as assessed by cell survival, propidium iodine staining, and transmission electron microscopy. Concomitantly with the apoptosis, there was the appearance of pl8 Bax-alpha, an apparently novel variant low molecular weight form of the p21 Bax-alpha found in normal cells. There was also a slight diminution in Bcl-xL, with a resultant drop in the Bcl-xL:Bax-alpha ratio. Treatment of cells with CD40-L partially inhibited the development of apoptosis in response to IFN-alpha. At the same time, generation of p18 Bax-alpha was reduced, which suggests that this plays a part in the apoptotic process. These findings may throw light on the development of lymphomas and perhaps point to future ways of improving therapy with IFN-alpha.

Diagnosis of peripheral lung cancer in cases of tumors 2 cm or less in size.

This study evaluates the results of preoperative diagnosis on 108 resected cases of peripheral lung cancer with a tumor size of 2 cm or less. Transbronchofiberscopic curettage was performed in 85 cases under x-ray TV fluoroscopy. Only 65 of 85 patients were positive on the initial bronchoscopy (76.5 percent), with repeated bronchoscopy, and 71 of 85 were positive for malignant cells (83.5 percent). The cases for which diagnosis could not be obtained with curettage were those in which the tumor shadow was not visible by x-ray TV fluoroscopy, those in which the lesions arose extramurally compressing the bronchi, or in cases of subpleural lesions. Curettage did not yield a diagnosis in cases with lesions less than 1.1 cm. In cases of small lung cancer lesions, peripheral transbronchofiberscopic curettage should be expected to yield a false-negative rate of approximately 15 percent. Therefore, even when the results of curettage are negative, further detailed examinations including needle biopsy, are necessary to obtain a definitive diagnosis.

Measurement of (90)Sr in environmental samples by cation-exchange and liquid scintillation counting.

A new method for the measurement of (90)Sr in environmental samples by cation-exchange and liquid scintillation counting is described. Strontium carbonate is purified by precipitation and ion-exchange, weighed for the determination of chemical yield, dissolved in hydrochloric acid and mixed with the liquid scintillator, Aquasol-2. Two channels of a low-background liquid scintillation counter are used to determine (90)Sr, (90)Y and (89)Sr, free from the effects of environmental tritium. The values of (90)Sr obtained by this method are in good agreement with those from ordinary (90)Y milking and the gas proportional counting method. The concentration of (90)Sr in the air at Tokai-mura in Japan has been measured by the new method.

Particulate and dissolved elemental loads in the Kuji River related to discharge rate.

In order to investigate influences of discharge rates on fluvial transport behavior of elements in the Kuji River, Japan, suspended and dissolved phase concentrations in the river water were measured together with the water flow rates. The concentrations of suspended particulate matter (SPM) increased by two or three orders of magnitude with the water discharge, and also had seasonal variations. Adsorptive elements or heavy metal elements, which tend to form insoluble compounds, were present generally as suspended forms in the river waters, and their dissolved form concentrations tended to increase with the water discharge. On the other hand, non-adsorptive elements such as alkali and alkaline-earth elements were present as dissolved forms under a normal flow rate condition, but equivalent quantity of suspended species also occurred under a high flow condition. In this case, the dissolved form concentrations decreased with the water discharge. Characterization of SPM with analyses of chemical compositions, scanning electron microscopy (SEM) and X-ray diffractometry (XRD) indicated that clay mineral such as montmorillonite would be the main carrier material of trace and major elements during the fluvial transport in the Kuji River. Variations of contents of selected elements in SPM with the water discharge indicated that river bottom sediments, which are probably potential sources of SPM added due to high water flow rates, would be different in chemical compositions from SPM floating under a normal flow condition. Variations of chemical compositions of SPM as well as of suspended or dissolved form concentrations of elements in the river waters were formulated as a function of the water discharge rates.

Lung cancer accompanied by erythema.

Skin manifestations associated with malignant diseases are designated syndroma dermatotumorale. A case of lung cancer combined with atypical erythema is reported. A 70-year-old man was admitted to hospital because of a 4-month history of atypical erythema of unknown origin. A nodule in the right lung was revealed on chest roentgenogram which was diagnosed as lung cancer. After right upper lobectomy, the erythema regressed gradually and disappeared completely in 7 days. It is suggested that the erythema was a manifestation associated with the lung cancer.

[Measurement of small quantities of alpha 2-macroglobulin in bronchoalveolar lavage fluid by enzyme-labelled immunoassay, and its clinical implication in pulmonary disease].

We established an enzyme labelled immunoassay for the determination of alpha 2 macroglobulin (alpha 2M). The assay range was from 2 to 140 ng/ml and the within-assay coefficient of variation (CV) were 5.2% at 31.2 ng/ml and 6.4% at 62.5 ng/ml. Between-day CV ranged from 6.9% to 15.4%. Using this method, alpha 2 M was determined in the bronchoalveolar lavage fluid (BALF) from patients with interstitial lung diseases. Those diseases were active and inactive sarcoidosis, hypersensitivity pneumonitis and idiopathic pulmonary fibrosis (IPF, including collagen disease). We divided the IPF patients into two groups, 'acute type' and 'chronic type', judging from the prognosis. alpha 2 M/Albumin ratio in BALF in the active sarcoidosis and acute type IPF groups is significantly higher than that in the inactive sarcoidosis and chronic type IPF. These findings suggest that alpha 2 M in BALF can be a sensitive marker of the interstitial lung disease.

[Pleuropneumonectomy with combined resection of diaphragma, superior vena cava, and pericardium, for invasive thymoma with pleural dissemination].

A 63-year-old female was admitted to our hospital with chief complaint of exertional dyspnea. Right massive pleural effusion was found. After chest drainage was performed, chest CT scan and MRI showed several masses in the anterior mediastinum and on the diaphragma. Mesothelioma was suspected for needle biopsy. Pleuropneumonectomy with combined resection of diaphragma, superior vena cava and pericardium was performed. Histologically, the Tumor was mixed thymoma. She is well and alive for three years after operation. Pleuropneumonectomy may be a choice of effective treatments for invasive thymoma with pleural dissemination.

[Continuous oximetry monitoring in patients undergoing home oxygen therapy for chronic respiratory failure].

We investigated the clinical usefulness of continuous nocturnal oxygen saturation monitoring in patients undergoing home oxygen therapy (HOT). The subjects were 11 patients with chronic respiratory disease in the process of healing from acute exacerbation. None were mechanically ventilated. Each subject underwent full overnight oximetry. One patient was excluded from further investigation because of periodic desaturation suggestive of sleep apnea. The remaining 10 subjects included 5 patients with sequelae of pulmonary tuberculosis, 2 with diffuse panbronchiolitis, 1 with chronic pulmonary emphysema, 1 with chronic bronchitis, and 1 with kyphoscoliosis. All underwent full overnight and 30 min daytime oximetry monitoring for 23.7 +/- 7.4 (mean +/- SD) consecutive days. Daytime oximetry was performed when subjects were awake and resting in supine position. Mean nocturnal oxygen saturation (NmSpO2) and mean daytime oxygen saturation (DmSpO2) were calculated from data obtained from 0:00 through 5:00 hrs and from data obtained during a stable 10 min daytime period, respectively. The difference between NmSpO2 and DmSpO2 (delta SpO2), the percentage of total sleep time with SpO2 < or = 90% (DST 90) and nocturnal lowest oxygen saturation (NLSpO2) were calculated once daily for each subject. There were significant (p < 0.05) correlations between NmSpO2 and NLSpO2, between NmSpO2 and DST 90, and between NLSpO2 and DST 90 in all subjects. However, significant (p < 0.05) correlations between NmSpO2 and DmSpO2 were observed in only 6. During acute exacerbation, NmSpO2 was lower than DmSpO2, and delta SpO2 increased. Conversely, with the amelioration of acute symptoms, delta SpO2 decreased and NmSpO2 was higher than DmSpO2. There was a significant (p < 0.05) reverse correlation between NmSpO2 and delta SpO2 in 9 subjects. We concluded that monitoring nocturnal oxygen saturation is clinically useful to assessments of oxygenation status in patients undergoing HOT, and that it may assist the early diagnosis of acute exacerbation of respiratory failure.

[Adenosine deaminase activity in bronchoalveolar lavage fluid of sarcoidosis patients].

We studied adenosine deaminase (ADA) activity in bronchoalveolar lavage fluid (BALF) specimens from 24 patients with sarcoidosis. Mean BALF-ADA activity was significantly (p < 0.0001) elevated in patients with sarcoidosis (1.02 +/- 1.01 IU/L, mean +/- SD) compared to the subjects in a healthy control group (0.08 +/- 0.29 IU/L). In the sarcoidosis patients with high BALF-ADA activity (> or = 1.0 IU/L), AaDO2 was significantly (p < 0.0001) elevated compared to its level in patients with normal BALF-ADA activity (< 1.0 IU/L). BALF-ADA activity was significantly (p < 0.01) higher in patients who exhibited lung-field accumulations on 67Ga scintigrams compared to those with no accumulations, and significantly (p < 0.001) higher in patients undergoing corticosteroid treatment compared to in those patients who did not receive such treatment. These findings were similar to the results of studies using data on BALF-ADA/albumin ratios. Furthermore, they suggest that the localized production of ADA may increase in sarcoidosis patients displaying 67Ga scintigram lung-field accumulations with increased AaDO2, and that BALF-ADA activity may be a useful indicator of disease activity and the need for treatment.

[Measurement of adenosine deaminase activity in bronchoalveolar lavage fluids as a tool for diagnosing miliary tuberculosis].

Measurement of the activity of adenosine deaminase (ADA) in pleural effusions has been reported to be useful in the diagnosis of tuberculous pleuritis. To determine whether ADA activity can also be used to diagnose miliary tuberculosis, it was measured in bronchoalveolar lavage fluid from 64 subjects: 6 patients with miliary tuberculosis, 21 patients with sarcoidosis, 15 patients with idiopathic interstitial pneumonia, 15 patients with other diseases, and 7 healthy controls. Mean ADA activity was 5.02+/-3.75 IU/L (mean+/-SD) in patients with miliary tuberculosis; 1.06+/-0.99 IU/L in patients with sarcoidosis; 0.21+/-0.43 in patients with idiopathic interstitial pneumonia; and 0.30+/-0.51 IU/L in healthy controls. The level in patients with miliary tuberculosis differed significantly from the others (p<01.01). All 6 patients with miliary tuberculosis, 3 patients with sarcoidosis, 1 patient with Wegener's granulomatosis, and 1 patient with lymphangitic carcinomatosis had levels of ADA activity in bronchoalveolar lavage fluid that exceeded 2.0 IU/L. With a cut-off value of 2.0 IU/L the sensitivity of this test to miliary tuberculosis is higher than that of other diagnostic aids. Measurement of ADA activity in bronchoalveolar lavage fluid may be of great value in the early diagnosis of miliary tuberculosis.

[A case of Fusobacterium necroforum sepsis].

A 26-year-old man was admitted to our hospital with a 10-day history of sore throat, high fever, and right knee joint pain. On physical examination, the pharynx was considerably inflamed, and the right knee joint was swollen and extremely tender. Chest radiography showed multiple, bilateral nodules and masses with pleural effusions. Fusobacterium necorforum grew from samples of blood, pleural effusion, and pus taken from the knee joint. The patient was treated with intravenous clindamycin, ventilatory support, and continuous chest and knee joint drainage. His condition progressively improved and he was discharged on the 66th hospital day. A disease caused by an oropharyngeal infection with secondary suppurative thrombophlebitis of the internal jugular vein, and complicated by multiple metastatic infections is called postanginal septicemia, or Lemierre syndrome. Before the discovery of antibiotics, this disease usually was fatal. The widespread use of antibiotics for treat oropharyngeal infections may have caused a number of reported cases. Lemierre syndrome is an uncommon complication of oropharyngeal infection, and it may be fatal if diagnosis is delayed. Careful attention must be directed to patients with oropharyngeal infection who have signs and symptoms that suggest metastatic infection.

[A case of sho-saiko-to-induced pneumonitis, diagnosed by lymphocyte stimulation test using bronchoalveolar lavage fluid].

A 66-year-old Japanese man was admitted to our hospital with fever, cough and dyspnea. He had been taking Sho-Saiko-to, a traditional Chinese medicine, for twenty days. On admission, chest X-ray examination revealed a reticular pattern in the bilateral lungs, and respiratory failure was evident. Serum levels of CRP and LDH were elevated. A differential cell count of the bronchoalveolar lavage fluid (BALF) showed that lymphocytes and eosinophils were increased. Alveolitis with lymphocyte and plasma cell infiltration was observed in a pathological specimen obtained by transbronchial lung biopsy. After all drugs had been discontinued, PaO2, serum CRP and serum LDH improved, and the reticular pattern in the bilateral lungs gradually resolved without administration of corticosteroids or antibiotics. A lymphocyte stimulation test for Sho-Saiko-to using BALF gave a positive result, although LST using blood gave a doubtful reaction. The diagnosis of Sho-Saiko-to-induced pneumonitis was made from the clinical course, laboratory findings, BALF cell analysis, pathological findings and LST using BALF. Only 13 cases of pneumonitis due to administration of traditional Chinese medical drugs have been reported. This case suggests that LST using BALF is useful for the diagnosis of drug-induced pneumonitis.

[Evaluation of interstitial shadows by thin slice computed tomography].

We developed a method for evaluation of interstitial shadows by thin slice computed tomography (CT). We studied thirty-seven cases with interstitial shadows confirmed by thin slice CT, and 13 normal cases. Thin slice CT was scanned on 3 levels: 1) at the aortic arch, 2) about 2 cm below the carina, and 3) about 1 cm above the right diaphragm. Each level was scanned bilaterally to produce 6 regions of interest (ROI); CT numbers were recorded from each ROI. CT numbers ranging from -1000 to 100 Hounsfield units (H.U.) were classified into 4 different bands, -1000 - (-900) H.U. (low band), -900 - (-750) H.U. (medium low band), -750 - (-500) H.U. (medium high band), and -500 - 100 H.U. (high band), for cases with interstitial shadows and for normal cases. We calculated the prevalence of CT numbers in each band for both normals and subjects with interstitial shadows. Compared to the normals, subjects with interstitial shadows showed fewer pixels in the medium low band and more pixels in the medium high and high bands. Furthermore, the prevalence of CT numbers in each band correlated with %TLC, %VC, and pulmonary diffusing capacity. We conclude that thin slice CT is a useful method for evaluation of the extent and characteristics of interstitial shadows.

Bcl-2 protein expression in lung cancer and close correlation with neuroendocrine differentiation.

For determination of the cellular distribution of bcl-2 expression in lung cancer and clarification of its correlation with cell neuroendocrine differentiation, Bcl-2 immunostaining was carried out on a large series of formalin-fixed, paraffin-embedded lung cancer samples, and four general neuroendocrine marker and seven peptide hormone stainings were carried out on all Bcl-2-positive squamous cell carcinomas and adenocarcinomas of the lung as well as on 8 pulmonary neuroendocrine carcinomas histologically diagnosed. In addition, 3 small cell lung cancer cell lines were studied by Western blotting. Neuroendocrine differentiation in Bcl-2-negative squamous cell carcinomas and adenocarcinomas was examined with chromogranin A and alpha-subunit of Go protein stainings. Bcl-2 protein was detected in 104/111 small cell carcinomas, 8/8 neuroendocrine carcinomas, 0/6 typical (well differentiated) carcinoids, 23/64 squamous cell carcinomas, 4/65 adenocarcinomas, and all 3 small cell lung cancer cell lines. All 8 neuroendocrine carcinomas, 11 of the Bcl-2-positive squamous cell carcinomas, and all 4 Bcl-2 positive adenocarcinomas expressed multiple neuroendocrine markers. The distributions of Bcl-2 and neuroendocrine marker immunoreactivity closely paralleled each other on consecutive sections. In squamous cell carcinomas, Bcl-2-positive cells could be roughly subdivided into those with neuroendocrine differentiation features, usually demonstrating intense Bcl-2 staining, with basaloid tumor cells usually expressing weak to moderate Bcl-2 staining. The present study clearly shows Bcl-2 protein expression to be remarkably differentially regulated according to histological types of lung cancers and to appear to quite likely be closely associated with neuroendocrine differentiation of tumor cells, indicating that bcl-2 is importantly involved in cell development and differentiation, in addition to protecting cells from apoptosis. Bcl-2 might be usable as a neuroendocrine marker in lung cancers and possibly also in neural-crest-derived tumors.

Involvement of TRAIL/TRAIL-R interaction in IFN-alpha-induced apoptosis of Daudi B lymphoma cells.

Interferon-alpha (IFN-alpha) exerts the anti-tumour effect on various tumours at least partly through induction of apoptosis. Apoptosis is induced by members of the tumour necrosis factor (TNF) family, including Fas (CD95) and TNF-related apoptosis-inducing ligand (TRAIL). In the present study, we examined whether the TRAIL/TRAIL-R system is involved in IFN-alpha-induced apoptosis using Daudi B lymphoma cells. IFN-alpha upregulated the expression of TRAIL within 12 h, as assessed by flow cytometry and RT-PCR, and the level increased with time until 72 h. The levels of both TRAIL-R1 and TRAIL-R2, low in Daudi cells, were enhanced by IFN-alpha. The enhanced TRAIL-R1/-R2 appeared to function as a death-inducing molecule since IFN-alpha-stimulated cells were more susceptible to TRAIL-induced cell death. The IFN-alpha-stimulated Daudi cells or their derived culture supernatants displayed cytotoxicity against TRAIL-sensitive, but not resistant lines. Moreover, the IFN-alpha-induced reduction in mitochondrial membrane potential preceding the induction of apoptosis was substantially prevented by neutralizing anti-TRAIL monoclonal antibody. Taken together, IFN-alpha-induced apoptosis appears to be mediated by the autocrine and/or paracrine loop involving TRAIL/TRAIL-R.

[A case of T-cell lymphoma showing multiple nodular shadows and an elevated titer of human T-lymphotropic virus type I (HTLV-I) antibody].

A 42-year-old man, born in Chiba prefecture, was admitted to our hospital because of multiple nodular shadows on chest X-ray film and an elevated titer of human T-lymphotropic virus type I (HTLV-I) antibody. The pulmonary lesion was diagnosed as T-cell lymphoma by open lung biopsy. There has been only one previous report of T-cell lymphoma showing multiple nodular shadows on chest X-ray. The elevated titer of HTLV-I antibody strongly suggested that the present case was one of adult T-cell lymphoma.

[Aniline-induced methemoglobinemia monitored by pulse oximetry].

A case of aniline-induced methemoglobinemia is reported. When the pulse oximeter reading (SpO2) was 80%, the oxygen saturation measured by a co-oximeter (SaO2) was 61.2%, the oxygen saturation calculated from PaO2 values was 98.9% and methemoglobin level was 38.8%. After methylene blue injection, methemoglobin level decreased gradually. With a decrease of methemoglobin level, SpO2 approached SaO2. If disparity between SpO2 and the oxygen saturation calculated from PaO2 values is noted, the presence of methemoglobinemia must be suspected. In clinical situations, the pulse oximeter permits the continuous noninvasive monitoring of oxygen saturation. It is necessary, however, to consider the potential errors in pulse oximetry.