Non-genomic inhibitory effect of glucocorticoids on activated peripheral blood basophils through suppression of lipid raft formation.

We investigated the non-genomic effects of glucocorticoids (GCs) on inhibition of plasma membrane lipid raft formation in activated human basophils. Human basophils obtained from house dust mite (HDM)-sensitive volunteers were pretreated with hydrocortisone (CORT) or dexamethasone (Dex) for 30 min and then primed with phorbol 12-myristate 13-acetate (PMA, 10 ng/ml) or HDM (10 µg/ml). The expression of CD63, a basophil activation marker, was assessed by flow cytometry. Membrane-bound GC receptors (mGCRs) were analysed by flow cytometry and confocal laser microscopy. Lipid rafts were assessed using a GM1 ganglioside probe and visualization by confocal laser microscopy. Pretreatment of basophils with CORT (10(-4) M and 10(-5) M) and Dex (10(-7) M) significantly inhibited CD63 expression 20 min after addition of PMA or HDM. The inhibitory effects of GCs were not altered by the nuclear GC receptor (GCR) antagonist RU486 (10(-5) M) or the protein synthesis inhibitor cycloheximide (10(-4) M) (P < 0·05). CORT coupled to bovine serum albumin (BSA-CORT) mimicked the rapid inhibitory effects of CORT, suggesting the involvement of mGCRs. mGCRs were detectable on the plasma membrane of resting basophils and formed nanoclusters following treatment with PMA or HDM. Pretreatment of cells with BSA-CORT inhibited the expression of mGCRs and nanoclustering of ganglioside GM1 in lipid rafts. The study provides evidence that non-genomic mechanisms are involved in the rapid inhibitory effect of GCs on the formation of lipid raft nanoclusters, through binding to mGCRs on the plasma membrane of activated basophils.

Phase II multi-institutional prospective randomised trial comparing S-1+paclitaxel with S-1+cisplatin in patients with unresectable and/or recurrent advanced gastric cancer.

BACKGROUND: A combination of S-1 and cisplatin has been shown to be effective with acceptable safety for the first-line treatment of far-advanced gastric cancer in Japan. This is the first randomised phase II trial to compare S-1+paclitaxel with S-1+cisplatin in this setting. METHODS: Patients with unresectable and/or recurrent advanced gastric cancer were randomly assigned to receive one of the two regimens: S-1 (40 mg m(-2) twice daily) on days 1-14 plus paclitaxel (60 mg m(-2)) on days 1, 8, and 15 of a 4-week cycle (S-1+paclitaxel) or S-1 (40 mg m(-2) twice daily) on days 1-21 plus cisplatin (60 mg m(-2)) on day 8 of a 5-week cycle (S-1+cisplatin). The primary end point was the response rate (RR). Secondary end points included progression-free survival (PFS), overall survival (OS), and safety. RESULTS: A total of 83 patients were eligible for safety and efficacy analyses. In the S-1+paclitaxel and S-1+cisplatin groups, RRs (52.3% vs 48.7%; P=0.74) and median PFS (9 vs 6 months; P=0.50) were similar. The median OS was similar in the S-1+paclitaxel and S-1+cisplatin groups (16 vs 17 months; P=0.84). The incidence of grade 3 or higher haematological toxicity was 19.0% with S-1+paclitaxel and 19.5% with S-1+cisplatin. The incidence of grade 3 or higher non-haematological toxicity was 14.2% with S-1+paclitaxel and 17.1% with S-1+cisplatin. CONCLUSION: S-1+paclitaxel was suggested to be a feasible and effective non-platinum-based regimen for chemotherapy in patients with advanced gastric cancer. Our results should be confirmed in multicenter, phase III-controlled clinical trials.

Expression of carbonic anhydrase 9, a potential intrinsic marker of hypoxia, is associated with poor prognosis in oesophageal squamous cell carcinoma.

Carbonic anhydrase 9 (CA9) is a protein to be upregulated under exposure to hypoxic conditions. Hypoxic conditions are known to be associated with resistance to chemotherapy and radiotherapy, and with poor cancer prognosis. We examined CA9 expression in surgical specimens from oesophageal squamous cell carcinoma (ESCC) patients (n=127) using immunohistochemistry and real-time RT-PCR. We also examined CA9 expression and cell proliferation in ESCC cell lines (TE-2, TE-8 and TE-15) and an immortalised human oesophageal cell line (CHEK-1) using real-time RT-PCR, Western blotting, ELISA and MTT assay. Immunohistochemistry, high expression of CA9 was found in 63 of the 127 primary tumour specimens and was correlated with poor outcome (P=0.0003) and more aggressive/less favourable clinicopathological parameters (tumour size (P=0.0235), tumour depth (P<0.0001), regional lymph node metastasis (P=0.0031), distant lymph node metastasis (P=0.0077), stage (P<0.0001) and blood vessel invasion (P=0.006)). In vitro, CA9 expression in cultured cells and culture medium was also induced by hypoxia (P<0.01). CA9 is correlated with poor prognosis and malignant phenotype in patients with ESCC, and was upregulated by hypoxia. It is suggested that control of CA9 expression might improve the effectiveness of chemotherapy and radiotherapy in ESCC.

Effect of perioperative steroid therapy on the postoperative course of patients with oesophageal cancer.

BACKGROUND: Perioperative steroid therapy is often used in oesophageal cancer surgery and we evaluate the effect of this therapy on the secretory leukocyte protease inhibitor levels in the lungs (a major antiprotease in the conducting airways) and postoperative course in oesophageal cancer patients. METHODS: Twenty-one patients operated on for oesophageal cancer in 2003-2004 were treated with perioperative steroid therapy (250 mg of methylprednisolone intravenously 1 h before the operation). Fifteen consecutive patients operated on in 2002 served as a control group. Secretory leukocyte protease inhibitor in bronchoalveolar lavage fluid and the postoperative course in the two groups were compared. RESULTS: The mortality rate was 0% and there was no significant difference in the morbidity rate between the two groups. Days of intubation and systemic inflammatory response syndrome were significantly shorter for the steroid group. The bronchoalveolar lavage fluid secretory leukocyte protease inhibitor level was significantly higher in the steroid group than in the control group on postoperative days 2 and 3. The secretory leukocyte protease inhibitor level on postoperative day 3 was remarkably lower for the patients intubated for > or = 5 days and for those with pulmonary complications. CONCLUSION: Perioperative steroid therapy increased the bronchoalveolar lavage fluid secretory leukocyte protease inhibitor level and reduced the days of intubation and systemic inflammatory response syndrome in patients with oesophagectomy.

Interferon-gamma and granulocyte colony-stimulating factor in bronchoalveolar lavage fluid after oesophagectomy.

BACKGROUND: Activated polymorphonuclear leucocytes play a pivotal role in pulmonary complications after oesophagectomy. A lot of inflammatory mediators including interferon-gamma and granulocyte colony-stimulating factor are reported to modify the life span of polymorphonuclear leucocytes. AIMS: In this study we investigated whether interferon-gamma and granulocyte colony-stimulating factor are associated with pulmonary complications after oesophagectomy. PATIENTS AND METHODS: We measured interferon-gamma and granulocyte colony-stimulating factor concentrations in bronchoalveolar lavage fluid of 37 patients who had undergone oesophagectomy and examined the relationship between these mediators and pulmonary complications. RESULTS: Pulmonary complications occurred in nine patients (24%, Pneum(+)). There was no significant difference in age, gender, preoperative comorbid conditions, tumour stage, operation method, operating time or blood loss between the Pneum(+) group and another 28 patients(Pneum(-)). Days until extubation were significantly increased in the Pneum(+) group than in the Pneum(-) group. Interferon-gamma (on postoperative day 2) and granulocyte colony-stimulating factor (on postoperative days 1-3) in bronchoalveolar lavage fluid were significantly increased in the Pneum(+) group than in the Pneum(-) group and granulocyte colony-stimulating factor was significantly correlated with days until extubation. CONCLUSIONS: Our results indicate that bronchoalveolar lavage fluid granulocyte colony-stimulating factor is associated with respiratory conditions after oesophagectomy and assaying it can be useful for predicting pulmonary complications.

An immunohistochemical study of TIMP-3 expression in oesophageal squamous cell carcinoma.

Tissue inhibitor of metalloproteinase-3 (TIMP-3) inhibits the activity of matrix metalloproteinase, which may play an important role in carcinoma invasion and metastasis. We have investigated the relationship between TIMP-3 reduction and clinicopathological factors in oesophageal squamous cell carcinoma (ESCC). We examined tissue specimens that had been removed from 90 patients with thoracic oesophageal cancer who had undergone surgery between 1983 and 2001. Immunohistochemical staining was performed by the standard streptavidin-biotin method. Immunostaining of TIMP-3 was seen in the cytoplasm of cancer cells and normal oesophageal epithelial cells, particularly in cells located in shallow areas of the tumour. TIMP-3 preserved (+), moderate (+/-), and reduced (-) cases accounted for 30, 27, and 33 of the 90 patients, respectively (33, 30, 37%). Significant correlations were observed between TIMP-3 expression and depth of tumour invasion (P=0.001), number of lymph node metastases (P=0.003), infiltrative growth pattern (P=0.003), and disease stage (P=0.005). The survival rates of patients with TIMP-3 (-) cancer were significantly lower than those of patients with TIMP-3 (+) and TIMP-3 (+/-) cancer (P=0.0003). The mean 5-year survival rates of patients with TIMP-3 (+), (+/-), and (-) were 50, 58, and 21%, respectively. In conclusion, decreased expression of TIMP-3 protein correlates with invasive activity and metastasis. This makes the prognosis for patients with cancer that has lost TIMP-3 significantly less favourable than that for patients with cancer that has maintained TIMP-3.

CT is useful for identifying patients with complicated appendicitis.

BACKGROUND AND AIMS: We often come across patients with complicated appendicitis (perforation, abscess formation, or peritonitis) and it is essential to get accurate and detailed information on these patients preoperatively. In this study, we investigated whether or not preoperative computed tomography is useful for identifying these patients. PATIENTS AND METHODS: Plain and intravenously-contrasted helical computed tomography was obtained preoperatively in 94 (75%) of 125 patients who underwent appendectomy. Twenty-eight (30%) of the 94 patients had complicated appendicitis (Compli(+) group). We compared clinical factors and computed tomography findings of the Compli(+) group with those of 66 other patients (Compli(-) group). RESULTS: There was no significant difference between the Compli(+) and Compli(-) groups in gender, white blood cell count, the present rate of an enlarged appendix, or appendicolith. Fat stranding and free fluid on computed tomography were significantly associated with complicated appendicitis by both univariate and multilogistic regression analysis. Fourteen (70%) of the 20 patients with fat stranding and free fluid on computed tomography had complicated appendicitis and only 1 (4%) of the 28 Compli(+) patients had neither fat stranding nor free fluid on computed tomography. CONCLUSION: Our study has indicated that fat stranding and free fluid on computed tomography are significant for complicated appendicitis and helical computed tomography is a powerful tool for identifying patients with complicated appendicitis preoperatively.

FAK overexpression is correlated with tumour invasiveness and lymph node metastasis in oesophageal squamous cell carcinoma.

Focal adhesion kinase (p125(FAK); 'FAK') is a tyrosine kinase that is localised to cellular focal adhesions and is associated with a number of other proteins, such as integrin adhesion receptors. We performed an immunohistochemical analysis of FAK protein expression to determine the relationship between FAK overexpression and clinicopathological factors in oesophageal squamous cell carcinoma (ESCC). We examined tissue specimens that had been removed from 91 patients with thoracic oesophageal cancer who had undergone surgery between 1983 and 2001. Immunohistochemical staining was performed by the standard streptavidin-biotin method. Seven human ESCC cell lines-TE-1, TE-2, TE-8, TE-13, TE-15, TT, and TTn-and one immortalized human keratinocyte cell line-HaCaT-were used in Western blot analysis. Immunostaining of FAK was seen in the cytoplasm of cancer cells, particularly in cells located in the invasive fronts of cancer nests. FAK overexpression was detected in 54 of the 91 patients (59.3%). Significant correlations were observed between FAK overexpression and cell differentiation (P=0.0057), depth of tumour invasion (P=0.0023), presence of regional lymph node metastasis (P=0.0097), number of lymph node metastases (P=0.0026), and disease stage (P=0.012). The survival rates of patients with FAK-overexpressing cancer were significantly lower than those of patients without FAK-overexpression cancer (P=0.006). The 5-year survival rate of patients without FAK overexpression was 69%, whereas that of patients with FAK overexpression was 38%. On Western blot analysis, FAK was expressed at a high level in TE-1, TE-8, TE-15, and TT cells, at a moderate level in TE-2 and TTn cells, and at a low level in TE13 and HaCaT cells. FAK phosphorylation at tyrosine 397 was demonstrated in proportion to the intensity of FAK in all cell lines except TE15 and HaCaT. In conclusion, FAK overexpression of ESCC was related to cell differentiation, tumour invasiveness, and lymph node metastasis. Consequently, patients with ESCC who had FAK overexpression had a poor prognosis.

Seven-day triple therapy with omeprazole, amoxycillin and clarithromycin for Helicobacter pylori infection in haemodialysis patients.

BACKGROUND: Triple therapy is accepted as the treatment of choice for Helicobacter pylori eradication, but there is no consensus on how long the therapy should be maintained in haemodialysis (HD(+)) patients. Our aims in this study were to evaluate the safety and efficacy of the 7-day triple therapy in HD(+) patients. METHOD: Forty-seven HD(+) and 55 HD(-) patients with dyspepsia underwent endoscopy. The prevalence of H. pylori was detected by Giemsa stain, followed by the urea breath test (UBT). H. pylori(+) patients were scheduled to undergo 7-day triple therapy and the success of eradication was investigated by UBT. RESULTS: Forty-five (44%) patients were positive for H. pylori. Forty of them underwent triple therapy and 39 (98%) patients completed the treatment. Eradication was successful in 32 (82%) and unsuccessful in 7 (18%) patients. There was no significant difference between these groups in age, gender, endoscopic findings or HD, and only previous treatment was significant for eradication failure by univariate and multivariate logistic regression analysis. Side effects were observed in 2 (15%) of 13 HD(+) and 3 (11%) of 27 HD(-) patients, and one HD(-) patient had to stop medication because of severe nausea and vomiting. The eradication rate was 93% (28/30) in patients without previous treatment. The triple therapy was unsuccessful in 7 patients, and 4 of them again underwent 7-day triple therapy, but all resulted in failure. CONCLUSIONS: Seven-day triple therapy is safe and effective for primary treatment of H. pylori infection in both HD(+) and HD(-) patients, but a new treatment is necessary for patients with previous treatment.

Expression of p53 protein related to smoking and alcoholic beverage drinking habits in patients with esophageal cancers.

In esophageal squamous cell carcinoma (SCC), we used immunohistochemical analysis to further elucidate the correlation of p53 protein expression with clinicopathological factors, as well as with risk factors, such as tobacco smoking, alcohol consumption and a family history of cancer, using odds ratios (ORs). The expression of p53 protein was demonstrated in 55.1% of 89 esophageal SCC cases examined by immunohistochemistry. The expression of p53 protein did not correlate with gender, age, histological grading, lymph node metastasis, or TNM stage. The prevalence of p53 expression was significantly higher in patients with multiple primary esophageal cancers (P<0.05). p53 expression did not correlate with prognosis in univariate survival analysis. The esophageal SCC in either smokers or alcohol users was 4.67-5.83 times more likely to express p53 protein, while the likelihood of p53 expression in patients who use both tobacco and alcohol was more than 14.0 times. However, a significant association was not found between p53 expression and a family history of cancer, this having an OR as low as 1.85. The expression of p53 protein did not correlate with clinicopathological factors and prognosis in univariate and multivariate survival analyses. In contrast, tobacco smoking and alcohol consumption were shown to be strongly associated with p53 mutations in esophageal carcinogenesis.

Expression of nitrotyrosine is associated with angiogenesis in esophageal squamous cell carcinoma.

BACKGROUND: In our previous study, it was suggested that nitrotyrosine, a product of nitrogen species, found in esophageal squamous cell carcinoma (ESCC), may contribute to the progression of esophageal cancer. MATERIALS AND METHODS: To clarify whether nitrotyrosine expression is associated with apoptosis and/or angiogenic factors in ESCC, we have analyzed the relationship between nitrotyrosine presence and the apoptosis-related proteins, Bcl-2 and Bax, or CD34, the marker of vascular endothelial cells, by an immunohistochemical approach. RESULTS: Nitrotyrosine was detected in 21 out of 55 esophageal cancers. The correlation between nitrotyrosine presence and Bcl-2, Bax expression or apoptotic index (AI) was not significant. In contrast, nitrotyrosine presence was significantly correlated with the microvessel density (MVD); nitrotyrosine-positive specimens tended to show a high MAD, while nitrotyrosine-negative specimens tended to be associated with a low MVD (p<0.05). CONCLUSION: Our data suggest that NO induces progression of esophageal carcinoma through its effect on angiogenesis, rather than its effect on tumor apoptosis.

Epidermoid cyst of an intrapancreatic accessory spleen--a case report.

We report an extremely rare case of a splenic epidermoid cyst of the pancreas in a 51-year-old Japanese male with no clinical symptoms. A cystic tumor of the pancreatic tail was detected incidentally by abdominal ultrasonography. The patient was referred to the Gunma University Hospital for further examination of the pancreatic tumor. Upon diagnosis of a benign cystic tumor, a distal pancreatectomy with splenectomy was performed. Microscopically, the multicystic tumor, which was surrounded by the splenic tissue, was located within the pancreatic tissue. The cysts were lined by non-keratinizing squamous epithelium. The diagnosis of an epidermoid cyst occurring in an intrapancreatic accessory spleen was confirmed. To our knowledge, this is the 4th case ever reported in the English literature.

Nitrotyrosine in esophageal squamous cell carcinoma and relevance to p53 expression.

We have examined the expression of nitrotyrosine, a marker of peroxynitrite formation, in 55 esophageal cancers by immunohistochemistry. Nitrotyrosine was detected in 21 of 55 (38.2%) esophageal cancers. Comparison of nitrotyrosine expression and the pathological findings showed that there was a significant association between the expression of nitrotyrosine and each of the depth of tumor invasion (P<0.01), occurrence of metastasis (P<0.05), pathological stage (P<0.01), smoking status (P<0.05) and alcohol intake (P<0.05). The survival rate of patients with nitrotyrosine-negative cancer was significantly higher than that of patients with nitrotyrosine-positive cancer (log-rank test, P<0.01). p53 was detected in 29 of 55 (52.7%) esophageal cancers, however, p53 expression did not correlate with nitrotyrosine expression. In conclusion, nitrotyrosine, a product of nitrogen species, is expressed in esophageal squamous cell carcinoma, which suggests that exogenous risk factors, such as tobacco and alcohol, through NO, are associated with carcinogenesis and progression of esophageal squamous cell carcinoma.

An immunohistochemical study of p16, pRb, p21 and p53 proteins in human esophageal cancers.

BACKGROUND: Cell cycle-associated proteins, p16, pRb, p21 and p53 are important in regulating the G1-S checkpoint in the cell cycle, and their functional alterations play key roles in carcinogenesis and cell proliferation. MATERIALS AND METHODS: We immunohistochemically examined the expression of p16, pRb, p21 and p53 proteins by using surgically resected tissues from 35 patients with primary esophageal cancers. RESULTS: In 35 esophageal cancers, the expressions of p16, pRb, p21 and p53 proteins were detectable in 4(11.4%), 25(71.4%), 11(31.4%) and 20(57.1%) respectively. Interestingly, 24 of 25 pRb positive cancers (96.0%) had negative p16, whereas three of ten pRb-negative cancers (30.0%) had high levels of p16 (p < 0.05). In 11 cases of p21 positive immunostaining, there was lymph node metastasis (pN1) in 9 (81.8%). CONCLUSIONS: These results suggest that abnormalities of p16 protein may be closely associated with the carcinogenesis or cell proliferation of esophageal cancers.

[Primary pulmonary cryptococcosis diagnosed by medical examinations in 12 patients].

Primary pulmonary cryptococcosis is thought to be relatively less common than other lung mycoses, but recently there has been an increase in reports of patients with this disease. Our report covers 12 cases of primary pulmonary cryptococcosis in which the diagnosis was based on medical examinations. The patients consisted of 11 men and 1 woman, aged 27 to 58 years. Only 3 exhibited subjective symptoms. Roentgenograms showed cavitating tumor shadows in the lungs of 2 of the patients with subjective symptoms, and nodular shadows with diameters of 1.0 to 2.8 cm in all patients without subjective symptoms, indicating the possibility of lung cancer. The disease was diagnosed in 3 patients on the basis of transbronchial biopsy findings, and in 9 on the basis of needle aspiration biopsy findings. One patient was diabetic, but the others did not exhibit malignancies or other immunocompromised states. Antifungal drugs significantly reduced or eliminated the nodular shadows in 10 patients. Because patients with primary pulmonary cryptococcosis frequently lack subjective symptoms, prompt diagnosis is critical, particularly in view of the need to distinguish the disease from lung cancer.

Functional expression of the ascofuranone-sensitive Trypanosoma brucei brucei alternative oxidase in the cytoplasmic membrane of Escherichia coli.

Trypanosome alternative oxidase (TAO) is the terminal oxidase of the respiratory chain of long slender bloodstream forms (LS forms) of African trypanosoma, which causes sleeping sickness in human and nagana in cattle. TAO is a cytochrome-independent, cyanide-insensitive quinol oxidase and these properties are quite different from those of the bacterial quinol oxidase which belongs to the heme-copper terminal oxidase superfamily. Only little information concerning the molecular structure and enzymatic features of TAO have been available, whereas the bacterial enzyme has been well characterized. In this study, a cDNA encoding TAO from Trypanosoma brucei brucei was cloned into the expression vector pET15b (pTAO) and recombinant TAO was expressed in Escherichia coli. The growth of the transformant carrying pTAO was cyanide-resistant. A peptide with a molecular mass of 37 kDa was found in the cytoplasmic membrane of E. coli, and was recognized by antibodies against plant-type alternative oxidases from Sauromatum guttatum and Hansenula anomala. Both the ubiquinol oxidase and succinate oxidase activities found in the membrane of the transformant were insensitive to cyanide, while those of the control strain, which contained vector alone, were inhibited. This cyanide-insensitive growth of the E. coli carrying pTAO was inhibited by the addition of ascofuranone, a potent and specific inhibitor of TAO ubiquinol oxidase. The ubiquinol oxidase activity of the membrane from the transformant was sensitive to ascofuranone. These results clearly show the functional expression of TAO in E. coli and indicate that ubiquinol-8 in the E. coli membrane is able to serve as an electron donor to the recombinant enzyme and confer cyanide-resistant and ascofuranone-sensitive growth to E. coli. This system will facilitate the biochemical characterization of the novel terminal oxidase, TAO, and the understanding on the mechanism of the trypanocidal effect of ascofuranone.

1a-docosahexaenoyl mitomycin C: a novel inhibitor of protein tyrosine kinase.

A series of derivatives of mitomycin C conjugated with various fatty acids at position 1a was synthesized and the effect of these compounds on protein kinase activities was evaluated. 1a-Docosahexaenoyl mitomycin C (DMMC) selectively inhibited protein tyrosine kinase (PTK) activity in the postnuclear fraction of v-src-transformed NIH 3T3 cells although neither derivatives conjugated with other fatty acids or docosahexaenoic acid or mitomycin C did not. DMMC inhibited the activity of calmodulin-dependent kinase III and protein kinase A very weakly, and only barely affected protein kinase C activity. DMMC also attenuated autophosphorylation of immunoprecipitated p60v-src irreversibly. The addition of thiol compounds to the reaction mixture reversed the inhibition by DMMC, suggesting that some thiol moiety of PTK protein might be involved. DMMC also inhibited kinase activity of p210bcr-abl immunoprecipitated from the lysate of K562 cells. These results indicate that DMMC is a novel inhibitor of PTK.

[Thoracoscopy with two bronchoscopes in 50 patients with pleural effusion of unknown origin].

When performing thoracoscopy in patients with pleural effusion of unknown origin, we used two bronchoscopes simultaneously, one for observation and one for biopsy. A total of 50 patients with pleural effusion of unknown origin were studied. In all of those studies, pleural effusion was exudative, lymphocyte-dominant, had a low level of adenosine deaminase, no malignant cells, and no tuberculosis or other bacteria in pleural effusion smears. Fourteen were out-patients. A catheter was inserted into the pleural space under local anesthesia, and 300 ml to 500 ml of pure oxygen was injected to create a pneumothorax. Two flexible fiberoptic bronchoscopes were used simultaneously, one for observation and one for biopsy. Approximately 1 hour after the examination, the out patients were able to return home. Lesions in the pleural cavity were found in 42 of these 50 patients, and histological diagnosis was possible in 46. This is a simple procedure with no major side effects. The equipment required is familiar to pulmonary physicians, and the diagnostic yield is high.

[The mode of excitation of cat motoneurons].

1. Effects of afferent nerve stimulation on the waveform of the IS-spike were studied intracellularly in 101 motoneurons of cats which were either anesthetized with sodium pentobarbitone or decerebrated at the intercollicular level. 2. In 78 motoneurons repetitive or double shock stimulation of afferent nerves (the L 7 or S 1 dorsal root, biceps-semitendinosus nerve and tibial nerve) could produce 2-4 kinds of IS-spikes, as distinguished by differences in waveform. In 34 motoneurons 2-3 small potentials were superimposed on the smallest IS-spike with progressively longer latencies to form a larger IS-spike. These potentials were thought as dendritic in origin because only the dendrites are known to be capable of exhibiting multiple spikes and in 14 motoneurons dendritic spikes were actually shown to exist. 3. In 13 motoneurons following a repetitive stimulation of a dorsal root there occurred an increase in excitability of the membrane generating the IS-spike (the IS-membrane) in the absence of any appreciable changes in the resting membrane potential. This would indicate that the IS-membrane is subject to plastic changes or the IS-spike itself also originates in dendrites which are known to exhibit such changes.