Apoptosis and proliferative activity in mature and immature teratomas of the mediastinum.

BACKGROUND: Mediastinal teratomas are the most frequent mediastinal germ cell tumor. Whereas mature teratomas are benign tumors, immature teratomas are malignant. The purpose of this study was to find characteristics that could be used to distinguish between the growth and prognosis of the two teratoma types. METHODS: Twenty-four mediastinal teratomas (18 mature and 6 immature) were examined for apoptosis by 3'-end labeling of DNA and stained immunohistochemically for proliferating cell nuclear antigen, Bcl-2, Bax, p53 protein, and alpha-fetoprotein (AFP) expression in formalin fixed, paraffin embedded specimens. RESULTS: AFP was expressed in both immature teratomas and mature teratomas. Whereas p53 protein was expressed by most teratomas, p53 gene mutation was observed in only one patient with an immature teratoma in which the same mutation occurred in all tumor tissue components tested. Bax protein expression was relatively diffuse in mature teratomas but was focally expressed in immature teratomas. Bcl-2 protein was expressed focally in both mature and immature teratomas. Although the proliferative index was significantly higher in immature teratomas compared with mature teratomas (P < 0.001), the apoptotic index (AI) was significantly higher in mature teratomas compared with immature teratomas (P < 0.05). All patients except one in this study remain alive and disease free after undergoing tumor resection. CONCLUSIONS: The relatively high AI in mature teratomas may be due to the overexpression of the p53 protein. In contrast, immature teratomas exhibited higher proliferative activity and lower rates of apoptosis, which may explain the more aggressive behavior of these tumors. However, patients with immature mediastinal teratomas have a good prognosis if the tumor is resected completely after chemotherapy.

Expression of vascular endothelial growth factor in thoracic sarcomas.

BACKGROUND: A body of data indicates that vascular endothelial growth factor (VEGF) expression by carcinomas is closely related to the prognosis of carcinomas. However, the relationship between VEGF expression and the prognosis of sarcomas is contradictory. METHODS: Tissue from 27 cases of thoracic sarcoma was analyzed immunohistochemically for VEGF expression while tumor vascularity was quantified using an antibody directed against endothelial CD34. The relationship between VEGF expression and the prognosis of patients with sarcomas was then evaluated semiquantitatively. RESULTS: The microvessel count in sarcomas with strong VEGF expression was significantly higher than that in sarcomas with absent or faint VEGF expression. The disease-free survival rates of sarcomas with strong VEGF expression were significantly lower than those of sarcomas with absent or faint VEGF expression. We found that strong VEGF expression impacted on the disease-free survival in multivariate analyses. CONCLUSIONS: VEGF expression of thoracic sarcomas is directly related to angiogenesis and tumor vascularity, and our findings suggest that strong VEGF expression is an independent prognostic factor in patients with thoracic sarcomas.

Microsatellite alterations in patients with thoracic sarcoma.

Few studies on sarcomas have examined the relationships between microsatellite alterations in particular loci, tumor prognosis and tumorigenesis, because sarcomas are uncommon and those prognoses can be confounded by coexisting factors, such as tumor site. We studied the relationship between microsatellite alterations and prognosis in 31 patients with thoracic sarcoma. The frequency of loss of heterozygosity (LOH) at 17p13 in stage IV sarcomas was significantly higher than that in stage I and III sarcomas (p<0.05). The 5-year survival for patients with LOH at 17p13 was significantly lower than that for patients without LOH (p<0.05). Six of 31 cases (19.4%) revealed replication error. These results suggest that p53 abnormality occurs during advanced stages of sarcoma and are related to patient prognosis, and it is possible that aberrations in mismatch repair activity are related to sarcoma tumorigenesis.

Increased extracellular ascorbate release reflects glutamate re-uptake during the early stage of reperfusion after forebrain ischemia in rats.

Ascorbate is highly concentrated in neuropils, and its extracellular release is closely related to that of the excitatory neurotransmitters. Thus, the extracellular release of ascorbate and glutamate was measured during the early stage of forebrain ischemia-reperfusion in the rat hippocampus using a microdialysis biosensor system. Male Wistar rats were anesthetized with halothane under mechanical ventilation and normothermia. Two probes of the microdialysis biosensor electrode were inserted in the hippocampus bilaterally. One probe was perfused with phosphate-buffered saline (PBS) and the oxidation signal of dialyzed ascorbate was recorded. A second electropolymerized probe was perfused with PBS containing glutamate oxidase for glutamate measurement. Forebrain ischemia-reperfusion was performed by bilateral carotid artery occlusion with hemorrhagic hypotension (MAP=30 mmHg) for 10 min (Group 10, n=10) or 15 min (Group 15, n=10), followed by reperfusion for 60 min. The release of glutamate increased significantly to 294% (Group 10) and 334% (Group 15) during ischemia, and then decreased rapidly. In Group 15, however, it remained significantly higher after reperfusion than in Group 10. The release of ascorbate increased significantly to 504% (Group 10) and 334% (Group 15) after reperfusion. In Group 10, it was significantly higher for 5-15 min after reperfusion than in Group 15. The marked increase of ascorbate during reperfusion was associated with the rapid decrease in glutamate. The extended time of ischemia significantly inhibited glutamate re-uptake and ascorbate release during reperfusion. These findings suggest the extracellular ascorbate release during reperfusion after global ischemia as a marker of glutamate re-uptake.

A simple HPLC method to determine urinary phenylmercapturic acid and its application to gasoline station attendants to biomonitor occupational exposure to benzene at less than 1 ppm.

The objective of this study was to establish a hand-saving method to measure phenylmercapturic acid (PMA) and to examine urinary PMA as a marker of occupational exposure to benzene at levels less than 1 ppm. A simple HPLC method was developed to analyse PMA by monitoring absorption at 195 nm of the ef? uent from an ODS-3 column with acetonitrile-methanol-perchloric acid-water as a mobile phase. The detection limit of the method was 0.2 µg l(-1) with sufficient reproducibility. The method was applied to end-of-shift urine samples from 70 gasoline station attendants exposed to up to 107 ppb benzene, and 20 non-exposed controls of both sexes. Time-weighted average (TWA) exposure to benzene was measured by diffusive sampling. A regression analysis was applied to examine the quantitative relationship between the intensity of exposure to benzene and PMA in the end-of-shift urine samples. Multiple regression analysis showed no effects of age, sex, smoking and co-exposure to toluene and xylenes on urinary PMA. There was a linear relationship between TWA benzene exposure and urinary PMA (r = 0.60-0.67, P < 0.01). Background PMA in urine of the non-exposed controls was low and scattering of PMA around the regression line was narrow so that those with 20 ppb benzene exposure can be separated from the non-exposed by urinalysis for PMA. Thus, urinary PMA is sensitive enough for biological exposure monitoring of those exposed to less than 1 ppm benzene.

Continuous real-time measurement of extracellular ascorbate release in the rat striatum in vivo during forebrain ischemia-reperfusion.

Apart from its physiological role as a major antioxidant, ascorbate is highly concentrated in neuropils and ascorbate-mediated protection from excitotoxins has been demonstrated in vitro. Therefore, extracellular release of ascorbate during the early stage of ischemia-reperfusion was measured using a microdialysis electrode technique. One or two probes of the microdialysis biosensor were inserted into the rat striatum. One probe (n=16) was perfused with phosphate-buffered saline (PBS) for continuous oxidative signal recording. A second electropolymerised probe inserted into the other side of the striatum was perfused with PBS containing ascorbate oxidase in six rats. Forebrain ischemia-reperfusion was performed for 10min, followed by reperfusion for 60min. Ascorbate increased transiently during ischemia, and markedly to a maximum of 247.5+/-55. 8 microM from the baseline of 68.5+/-25.3 microM after reperfusion. The marked increase of extracellular ascorbate may be a marker of the early stage of reperfusion.

Undifferentiated spindle-cell sarcoma of the chest wall with vascular endothelial growth factor expression: report of a case.

The expressions of some growth factors have been immunohistochemically confirmed in several kinds of tumors, and in particular the expression of vascular endothelial growth factor (VEGF) has been reported to be closely related to tumor cell proliferation. We report herein a case of undifferentiated spindle-cell sarcoma arising in the chest wall with VEGF expression. A 67-year-old man, who presented with coughing, was found to have an abnormal shadow on his right chest wall. He was admitted to Chiba Rosai Hospital and preoperative diagnosis of the tumor was sarcoma. The tumor was thus resected along with the right chest wall and right lower lobe of the lung. Histopathologically, the tumor cells were spindle-shaped and showed severe atypism. The tumor cells were positive for vimentin and VEGF antibody with immunohistochemical staining, but they did not show differentiation to any special type of sarcoma. The tumor was diagnosed to be undifferentiated spindle-cell sarcoma. The microvessel density of the tumor was measured using CD34 and it was found to be higher than the average density of usual sarcomas. The prognosis of this case was poor. The patient died of tumor metastasis to the lung and bone 1 year later in spite of the fact that the tumor was resected.

Castleman's disease in the posterior mediastinum: report of a case.

We report a case of Castleman's disease which developed in the posterior mediastinum, with a review of the Japanese literature. A 19-year-old female patient with asthma was pointed out to have an abnormal shadow in the right posterior mediastinum on chest X-ray. We had tried to perform thoracoscopic surgery for this tumor, but we had to convert the surgical approach from thoracoscopy to a thoracotomy because of both tight adhesion of the tumor and muscle and profuse bleeding from the tumor. The tumor was diagnosed to be the hyaline vascular type of Castleman's disease histopathologically. In the Japanese literature, Castleman's disease, which develops in the posterior mediastinum, has been reported to often accompany tight adhesion between the tumor and surrounding tissue, and profuse bleeding thus cannot be avoided at surgery. The large amount of bleeding observed during the surgery of a patient with Castleman's disease in the posterior mediastinum may be due to tight adhesion and hypervascularity of the tumor. Therefore, care should be exercised in choosing the surgical approach if Castleman's disease is suspected in cases of posterior mediastinal tumor.

Effects of intraluminal trypsin and bile on the exocrine and endocrine pancreas after pancreaticobiliary diversion and biliodigestive shunt.

Pancreaticobiliary diversion (PBD) and biliodigestive shunt (BDS) cause long-standing hypercholecystokininemia followed by pancreatic hyperplasia. These changes have been suggested to be due to the lack of intraluminal trypsin and bile, respectively, in the upper small intestine. The aim of these experiments was to study the effect of restoration of intraluminal trypsin and bile on plasma levels of cholecystokinin (CCK) and the changes found in exocrine and endocrine pancreas after PBD and BDS. Male Sprague-Dawley rats were used. PBD was done in 16 rats, eight of which had trypsin dissolved in 50 mM sodium bicarbonate (SB), and eight had SB only by gastric intubation twice daily. BDS was done in another 16 rats, eight of which had bile dissolved in SB, and eight had SB in a similar manner. Sham-operated rats had SB and served as controls. After 4 weeks, the rats were killed, and the concentrations of circulating CCK, gastrin, glucose, glucagon, and insulin were determined. The pancreas was removed, weighed, and analyzed for contents of water, protein, and DNA. In another study, PBD-operated rats got trypsin in varying dosages or trypsin and taurocholate in combination for 2 weeks before death. The concentrations of plasma CCK and glucagon were elevated after both PBD and BDS. PBD decreased the concentration of gastrin in plasma. PBD caused an increase of pancreatic weight and the contents of protein and DNA. Trypsin substitution to PBD-operated rats did not affect plasma CCK or glucagon levels, but the PBD-induced increases in weight and DNA content were counteracted by trypsin. Higher dosages of trypsin did not further influence the effects seen after PBD. Pancreatic weight and DNA content were increased after BDS. Bile administration completely abolished the increase in plasma CCK and glucagon, as well as the gain in pancreatic weight, and reduced the increase in pancreatic DNA. Substitution with bile to BDS-operated rats abolished the increase in the plasma levels of CCK and glucagon, as well as the trophic effects on the pancreas. Trypsin substitution to PBD-operated rats partly reversed the trophic effects on the pancreas but not the hormonal changes in plasma. Thus the trophic effects on the pancreas exerted by BDS seem to be dependent on the lack of bile in the upper small intestine, whereas the effects of PBD only partly are a consequence of the absence of intraluminal trypsin.

Regulation of the activity and polymerization status of recombinant human cytosolic thymidine kinase by thiols and ATP.

The cDNA clone encoding human thymidine kinase (hTK), was expressed in E. coli using a prokaryotic expression vector, pKK 223-3. The kinetics of the recombinant hTK (rhTK) were similar to those of cytosolic TK but not of mitochondrial TK. rhTK was highly purified in the presence of either ATP or dithiothreitol (DTT). The specific activity of rhTK purified in the presence of ATP [rhTK(ATP)] was lower than that of rhTK purified in the presence of DTT [rhTK(DTT)]. Activity of the purified rhTK(ATP) was enhanced by addition of thiols including DTT, cysteine, homocysteine and beta-mercaptoethanol but inhibited by various sulfhydryl reagents such as 5,5'-dithio-bis(2-nitrobenzoic acid). Hence, it was suggested that rhTK is a thiol-type enzyme. Apparent Mr of purified rhTK(ATP) was 100 kDa, which corresponds to the size of a tetramer (25 kDa subunit), while that of purified rhTK(DTT) was 50 kDa, the size of a dimer. The tetramer form of rhTK(ATP) was converted to the dimer by replacement of ATP by DTT. On the other hand, the dimer form of rhTK(DTT) was converted to the tetramer by addition of ATP. Thus, the catalytic activity of human cytosolic TK might be regulated by thiols as well as ATP via its polymerization status.

The primary mediastinal growing teratoma syndrome.

We encountered a case of mediastinal immature teratoma which revealed the feature of the so-called growing teratoma syndrome. A 20-year-old male with a cough was discovered to have an abnormal shadow in the mediastinum. The serum AFP was elevated to 3600 ng/ml. The specimen with percutaneous needle biopsy revealed mature teratoma. The tumor was suspected to be mature teratoma with a malignant component because of the high level of serum AFP and he underwent chemotherapy. The serum AFP declined markedly but the tumor further enlarged. The resected tumor was diagnosed as immature teratoma, although most of the tumor tissue was mature component.

Effects of nitric oxide synthase inhibition on extracellular glutamate and cerebral blood flow during forebrain ischemia-reperfusion in rat in vivo.

PURPOSE: To evaluate factors involved in global forebrain ischemia-reperfusion, the effects of the systemically administered NOS inhibitor, N(G)-nitro-L-arginine methyl ester (L-NAME), on changes in extracellular glutamate and cerebral blood flow (CBF) were studied during the early period of global forebrain ischemia-reperfusion, simultaneously measuring the glutamate released in the rat forebrain cortex and cortical CBF. METHODS: After injection of saline or L-NAME, forebrain ischemia-reperfusion was performed by bilateral carotid artery occlusion with controlled hemorrhagic hypotension (30 mmHg) for 10 min and reperfusion for 60 min. The microdialysis electrode and laser Doppler flowmetry were used for real-time monitoring of glutamate and CBF, respectively. RESULTS: During ischemia, glutamate increased linearly to over 100 muM and remained elevated 30 min after reperfusion in L-NAME-treated rats. In L-NAME-treated rats, CBF also remained significantly lower than baseline for 30-60 min after reperfusion, and glutamate was higher than in saline-treated rats throughout the experiment. CONCLUSION: A remarkable linear increase in glutamate release was observed during ischemia. L-NAME did not prevent this dramatic glutamate accumulation, and moreover, its level increased during reperfusion. The decrease in CBF response after reperfusion might be a factor of the elevated glutamate after reperfusion due to a decrease in reuptake of glutamate.

Descending necrotizing mediastinitis: report of a case.

A 47-year-old man was admitted to our hospital for treatment of an odontogenic infection. He presented with a fever, signs of sepsis, and neck swelling, and was initially diagnosed as having a neck abscess. After cervical drainage, he showed no improvement, and mediastinitis was detected by chest X-ray and computed tomography. A thoracotomy and mediastinal drainage was subsequently performed for descending necrotizing mediastinitis, which resulted in marked improvement. To date, only 83 cases of descending necrotizing mediastinitis have been reported in Japan. We present herein an additional case, followed by a review of the Japanese literature.

Surgical resection combined with intrathoracic hyperthermic perfusion for thymic carcinoma with an intrathoracic disseminated lesion: a case report.

Thymic undifferentiated carcinoma has a poor prognosis. We encountered a patient with thymic carcinoma associated with an intrathoracic disseminated lesion, who underwent surgery combined with intrathoracic hyperthermic perfusion after systemic chemotherapy and showed good results. The 45-year-old man was diagnosed as having a thymoma with an intrathoracic disseminated lesion. After he underwent three courses of systemic chemotherapy, he was admitted to our hospital. An anterior mediastinal tumor and an intrathoracic disseminated lesion remained, and were treated by surgical resection combined with intrathoracic hyperthermic perfusion. The tumors were histopathologically diagnosed as thymic undifferentiated carcinomas with pleural dissemination. At present, approximately 16 months after surgery, the patient is alive without recurrence.

Human pulmonary dirofilariasis: report of six cases.

We report six cases of pulmonary dirofilariasis diagnosed at our laboratory with clinical and pathological features. The nodules of dirofilariasis were round in three cases as previously reported, however dumbbell-shaped in two cases. The nodule did not attach to the pleura in four cases. Microscopically, the nodules were granulomas composed of central coagulation necrosis and peripheral fibrosis with round cell infiltration, histiocytes, and multinucleated giant cells. Necrotic pulmonary artery with single or multiple sections of degenerated nematode was observed in the center of the nodule. Dilated bronchioles with inflammation were observed in the nodule in four cases. Collapse of the alveoli, organizing pneumonia, hemosiderin-laden macrophages were observed around the nodule. We suppose that the nodule is not an infarction but a granuloma caused by antigen released from the nematode. Because the pulmonary dirofilariasis is difficult to be differentiated from primary or metastatic lung carcinoma, and the inflammation exists around the nodule, the nodule should be removed surgically.

Local cortical cerebral blood flow and response to carbon dioxide during anesthesia in patients with moyamoya disease.

PURPOSE: The CO(2) reactivity of cortical cerebral vessels and local cortical blood flow (l-CoBF) were evaluated during anesthesia in patients with moyamoya disease who were undergoing revascularization surgery. METHODS: Using laser-Doppler flowmetry, the CO(2) reactivity of cortical cerebral vessels and l-CoBF were measured continuously in five patients at the local surgical field of the middle cerebral artery (MCA) territory. RESULTS: Local-CoBF values obtained during the normocapnic condition varied from site to site of gyrus in the MCA region (0-73 ml. 100 g(-1).min(-1)). Local-CoBF was maximal at 39-43 mmHg of the PaCO(2) range, and decreased above and below this range. The response of l-CoBF to CO(2) was larger at the sites where the maximal level was obtained during normocapnia. In two patients, l-CoBF decreased by about 50%, and remained law even 40 min after administration of acetazolamide. CONCLUSION: In patients with moyamoya disease, l-CoBF values obtained during the normocapnic condition varied from site to site of gyrus, and not only hypocapnia but also hypercapnia decreased l-CoBF within the MCA region.

[Detection of emphysematous bullae in spontaneous pneumothorax by three-dimensional computed topography].

The patient was a 20-year-old woman with spontaneous pneumothorax, who had had pneumothorax twice on each side. Three-dimensional computed topographic (CT) images were made as follows. The range 12.5 cm caudad from the pulmonary apex was scanned by rapid helical CT, and three-dimensional images were obtained with the surface-rendering technique at a voxel extraction threshold ranging-850 HU to 1,600 HU. The three-dimensional CT images provided a stereotactic rendering of the lung surface. Irregular findings were observed at both pulmonary apices; they indicated the presence of one emphysematous bulla in the right lung and many emphysematous bullae of various sizes in the left lung. These findings were highly consistent with the sites and structures of the emphysematous bullae, as observed during thoracoscopy, which suggests that three-dimensional CT is a useful method for diagnosing and evaluating emphysematous bullae in patients with spontaneous pneumothorax.

[Hyperbaric oxygen therapy for radiation-induced hemorrhagic cystitis].

BACKGROUND: Radiation therapy has widely been used for cancers in the pelvis. Radiation cystitis, one of the late complications, presents often as hemorrhagic cystitis, which is refractory to the conventional therapy and may threaten the patient's life. We used hyperbaric oxygen therapy on patients with radiation cystitis to test its potential benifit. METHODS: Ten patients aged from 46 to 81 years with a mean of 62 years underwent one or more courses of hyperbaric oxygen therapy according to their symptoms, consisting of 20 sessions (3 to 5 sessions a week) at the Department of Hyperbaric Medicine, the University of the Ryukyus Hospital in the 9-year period from 1985 to 1994. They included 8 patients having a history of cervical cancer, one with external genital cancer and one with vaginal cancer. During the 75 min hyperbaric oxygen therapy patients received 100% oxygen at 2 absolute atmosphere pressure in the Multiplace Hyperbaric Chamber. RESULTS: Hematuria subsided and subjective symptoms including urinary frequency improved in seven patients. Cystoscopic findings including mucosal edema, redness, and capillary dilation were partially improved. The procedure subjectively and objectively palliated the 10 patients in a favorable manner. CONCLUSIONS: To date we have not armed any active procedure to control radiation-induced refractory hemorrhagic cystitis in terms of efficacy, invasiveness, and adverse effects. Therefore, in consideration of our clinical results, hyperbaric oxygen therapy appears to be useful for radiation cystitis.

[The effect of hyperbaric oxygenation on nitrite and nitrate production in rats treated with endotoxin].

The effect of hyperbaric oxygenation (HBO; 3 ATA, 90 min) on production of stable endproducts of nitric oxide (NO); nitrite (NO2-) and nitrate (NO3-), in plasma was studied in anesthetized rats treated with lipopolysaccharide (LPS). Intravenous injection of LPS (1 mg.kg-1) increased NO2-/NO3- production in plasma significantly 5-6 hrs after treatment, compared with non-treated rats. Production of NO2-/NO3- was not influenced by HBO performed 3 hrs after LPS injection. HBO performed 1 hr after LPS treatment, however, depressed NO2-/NO3- production significantly, compared with HBO performed 3 hrs after LPS, although NO2- production remained increased. These data suggest that HBO may influence the NO2-/NO3- production in NO producing processes in rats treated with LPS.