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Biomedical subjects

T Funatsu

Publications and source records attributed to T Funatsu.

At least 37 records · Page 2Linked to original sources
PubMed1996

[Flow cytometric analysis of the DNA content of resected non-small cell lung cancer with reference to long-term follow-up].

We measured the cellular DNA content of paraffin-embedded tumor specimens by flow cytometry from 340 cases of resected non-small cell lung cancer, and investigated the correlation of DNA content and prognosis of these cases with long-term follow-up. These 340 cases were divided into some populations according to pathological stage, histologic type, surgical curativity and N factor, and we compared the prognosis of DNA diploidy cases and DNA aneuploidy cases in each population. DNA aneuploidy cases had a significantly less favorable prognosis than DNA diploidy cases in population of stage I adenocarcinoma, stage IIIA non-small cell lung cancer and N2 cases among stage IIIA non-small cell lung cancer, all after curative operation. But in other populations, there was no significant difference in prognosis between DNA diploidy cases and DNA aneuploidy cases. In conclusion, DNA ploidy pattern is a prognostic factor for survival in patients with stage I adenocarcinoma and N2 cases of stage IIIA non-small cell lung cancer.

Adenocarcinoma↗
PubMed1996

[T cell lymphoma presenting as recurrent bilateral pulmonary infiltrates over five years].

A 52-year-old man presented with BOOP-like recurrent bilateral pulmonary infiltrates. In 1989 a chest X-ray film showed an infiltrative shadow in the right S6 region which disappeared after administration of prednisolone. Thereafter, bilateral patchy infiltrates recurred many times, and each time they resolved rapidly with steroid therapy. Skin eruptions on the face recurred. In October 1994 the patient underwent an open-lung biopsy of the infiltrate in the left S6 region. The pathological findings were consistent with BOOP, except for the moderate-to marked infiltration of lymphocytes. A lymphoproliferative disorder was suspected, and Southern blot analysis of the specimen revealed a rearrangement of the TCR-beta gene, which led to the diagnosis of T cell lymphoma. Ten months after the diagnosis, no recurrence of the lymphoma had been detected. In this case a gene analysis of the biopsy specimen was very useful for the diagnosis of T cell lymphoma.

Cryptogenic Organizing Pneumonia↗
PubMed1995

Imaging of single fluorescent molecules and individual ATP turnovers by single myosin molecules in aqueous solution.

Visualization of single actin filaments by fluorescence microscopy led to the development of new in vitro assays for analysing actomyosin-based motility at the molecular level. The ability to manipulate actin filaments with a microneedle or an optical trap combined with position-sensitive detectors has enabled direct measurements of nanometre displacements and piconewton forces exerted by individual myosin molecules. To elucidate how myosin generates movement, it is necessary to understand how ATP hydrolysis is coupled to mechanical work at the level of the single molecule. But the most sensitive microscopic ATPase assay available still requires over 1,000 myosins. To enhance the sensitivity of such assays, we have refined epifluorescence and total internal reflection microscopies to visualize single fluorescent dye molecules. We report here that this approach can be used directly to image single fluorescently labelled myosin molecules and detect individual ATP turnover reactions. In contrast to previously reported single fluorescent molecule imaging methods, which used specimens immobilized on an air-dried surface, our method allows video-rate imaging of single molecules in aqueous solution, and hence can be applied to the study of many types of enzymes and biomolecules.

Adenosine Triphosphate↗
PubMed1994

Single-molecule analysis of the actomyosin motor using nano-manipulation.

The elementary events in energy transduction by the actomyosin motor, driven by ATP hydrolysis, were directly recorded from multiple and single molecules using a recently developed technique for nano-manipulation of single actin filaments by a microneedle. In order to avoid the effects of random orientation of myosin and association of myosin with an artificial substrate in the surface motility assay, we used single myosin-rod cofilaments with various ratios. Distinct actomyosin attachment, force generation (the power stroke) and detachment events were detected at a very low myosin: rod ratio. At high load, one power stroke generated 5-6 pN peak force and 2.3 pN force averaged over the cycle, which were compatible with those deduced from noise analysis of force fluctuations caused by multiple molecules. As the load was reduced, the length of the power stroke increased. At near zero load, the length of a power stroke was approximately 17 nm. The results suggested that an ATPase cycle produces one power stroke at high load and many ones at low load.

Actins↗
PubMed1994

Structural and functional reconstitution of thin filaments in skeletal muscle.

Thin filaments were reconstituted by incorporating exogenous actin, tropomyosin and troponin into glycerinated skeletal muscle fibres or myofibrils. Firstly, thin filaments except short fragments at the Z line were selectively removed by treatment with plasma gelsolin, an actin severing protein. As a result, the fibres (or fibrils) lost the ability to generate active tension. Next, actin filaments were reconstituted by adding purified G-actin which polymerizes onto the actin fragments which remained at the Z line. Rhodamine phalloidin staining of myofibrils showed that exogenous actin was incorporated into the position where the intrinsic thin filaments located. Thin section electron micrographs of fibres showed that reconstituted actin filaments ran from the Z line to the inside of the A band, with some reaching the H zone. The number density of reconstituted actin filaments in the A band was about 20% of that found in intact fibres. The actin filament-reconstituted fibres (or fibrils) generated active tension in a Ca(2+)-insensitive manner and the tension was reversibly suppressed by 2,3-butanedione 2-monoxime. The recovered active tension was about 20% of tension developed by intact fibres. These results indicate that reconstituted actin filaments bear active tension similar to that borne by intact thin filaments. Thin filament-reconstituted fibres, which were prepared by adding purified tropomyosin-troponin complexes into actin filament-reconstituted fibres, showed Ca(2+)-sensitive tension generation. The maximum tension generated was not affected by the presence of tropomyosin and troponin. SDS-PAGE analysis showed that more than 25% of actin and 20% of tropomyosin and troponin was incorporated into the reconstituted fibres. These results indicate that the structure and function of thin filaments are substantially reconstituted by self-assembly of actin, tropomyosin and troponin. The reconstituted fibres and fibrils will be useful for studying the molecular mechanism of muscle contraction and its regulation.

Actin Cytoskeleton↗
PubMed1994

[A case of intractable pulmonary tuberculosis complicated by idiopathic thrombocytopenic purpura (ITP)].

The authors describe a case of pulmonary tuberculosis with multiple drug resistance and complicated by severe idiopathic thrombocytopenic purpura (ITP). The patient was initially treated with high-dose gamma globulin therapy and splenectomy. These procedures resulted in improvement of the patient's condition; the platelet count increased from 7,000/mm3 to 230,000/mm3 and the bleeding time fell from 15 min to 1 min 30 s. Given the significant improvement in the patient's condition, it was considered safe to carry out left pulmonary upper lobectomy and thoracoplasty. There have been no reports describing pulmonary resection performed for inflammatory lung disease in patients with ITP. High-dose gamma globulin therapy rapidly brings about an increase in the platelet count and a reduction of the bleeding time, and it thus extremely useful as preoperative treatment in surgical cases complicated by thrombocytopenia.

Humans↗
PubMed1994

[Diagnosis of mediastinal lymph node metastasis in primary lung cancer: comparing CT scan and mediastinoscopy].

In 105 patients with primary lung cancer who were operated on, the rate of diagnosis of mediastinal lymph node metastasis was determined, and the results of CT scanning and mediastinoscopy were compared. The rate of accuracy of mediastinoscopy was 93.8%, considerably higher than that of CT, which was less than 85%. For the lymph nodes other than #7, which indicate contralateral metastasis, the rate of accuracy of mediastinoscopy was even better and reached 96.3%. In the 9 patients who were proved by mediastinoscopy to have contralateral lymph node metastasis, the rate of accuracy of CT was only 44%. Mediastinoscopy was very effective in the diagnosis of mediastinal lymph node metastases from primary lung cancers.

Humans↗
PubMed1994

Preoperative mediastinoscopic assessment of N factors and the need for mediastinal lymph node dissection in T1 lung cancer.

The significance of preoperative N factor assessment in T1 lung cancer and the need for mediastinal node dissection in T1 N0 M0 cases were investigated. The results of mediastinoscopy were evaluated in patients with T1 adenocarcinoma or squamous cell carcinoma who underwent preoperative mediastinoscopy and thoracotomy from 1971 to 1991 (n = 164). Mediastinoscopy gave true negative results in 90% of patients, false negative in 1%, and true positive in 9%. The 5-year survival was 90% for patients with T1 N0 M0 disease who underwent nonradical dissection (n = 64) and 70% for those who underwent radical dissection (n = 61), indicating that the prognosis was significantly better (p < 0.05) with nonradical dissection. Distant metastasis was a common cause of death, and no death was related to local recurrence, whether nonradical or radical dissection had been performed. The results strongly suggest that preoperative mediastinoscopy and intraoperative node sampling are sufficient for assessment of N factors in T1 lung cancer. The possible relation between immunologic functional changes associated with mediastinal lymph node dissection and the prognosis in patients without evidence of positive lymph nodes should be clarified by a prospective randomized study.

Adenocarcinoma↗
PubMed1994

[A case of solitary plasmacytoma of the chest wall].

A 58-year-old man with a solitary plasmacytoma of the chest wall is reported. He was admitted because of a painless tumor on the right lateral chest wall. The chest CT scan showed a chest wall tumor surrounding the 7th and 8th ribs without rib destruction. A transcutaneous needle biopsy of the tumor with a Sure-Cut-Needle revealed a plasmacytoma. The bone marrow biopsy findings were normal. Under a diagnosis of solitary plasmacytoma, the chest wall tumor was resected. The tumor was an extramedullary plasmacytoma. Southern blot analysis of the immunoglobulin light chain gene of the tumor cells detected a rearrangement of genes in the lambda chain, while no rearrangements of immunoglobulin genes were detected in the bone marrow specimen. Seven months later another solitary plasmacytoma was found at the left radius, and was resected. Southern blot analysis of the immunoglobulin light chain gene was useful in the differential diagnosis from systemic myeloma and in determining the monoclonality of the tumor.

Blotting, Southern↗
PubMed1994

[Preoperative staging in T1 and T2 lung cancers by mediastinoscopy and mediastinal lymph node dissection].

Between 1975 and 1992, mediastinoscopy and thoracotomy were performed on 184 T1 and 271 T2 lung cancer cases consisting of adenocarcinoma and squamous cell carcinoma. Mediastinoscopy gave true negative findings in 90.8% of the T1 patients, true positive findings in 7.6% and false negative findings in 1.6%. The comparable rates were 76.7%, 17.0% and 6.3% in T2 patients. The 5-year survival rate was 91.3% for T1N0M0 patients (n = 64) who underwent non-radical dissection (= NRD), and 69.4% for those (n = 70) who underwent radical dissection (= RD). The rate with NRD was significantly better (p < 0.006). The 5-year survival rate was 63.2% for T2N0M0 patients (n = 62) undergoing NRD, and 49.8% for those (n = 72) undergoing RD, but the difference was not significantly. Distant metastasis was a common cause of death, whereas there were no deaths due to local recurrence in the T1N0M0 patients, whether NRD or RD was performed. These results support our opinions that preoperative mediastinoscopy and intraoperative node staging are sufficient for assessment of the N factor in T1 and T2 lung cancer, and that mediastinal node dissection should not be performed in T1N0M0 patients.

Adenocarcinoma↗
PubMed1993

Changes in mobility of chromaffin granules in actin network with its assembly and Ca(2+)-dependent disassembly by gelsolin.

As a final stage of cell signal transduction, secretory cells release hormones by exocytosis. Before secretory granules contact with the cell membrane for fusion, an actin-network barrier must dissociate as a prelude. To elucidate dynamical behaviors of secretory granules in actin networks, in vitro assembly and disassembly processes of actin networks were examined by means of dynamic light-scattering spectroscopy. We studied actin polymerization in the presence of chromaffin granules isolated from bovine adrenal medullas and found that the entanglement of actin filaments rapidly formed cages that confined granules in them. We also studied the effect of gelsolin, one of actin-severing proteins, on the network of actin filaments preformed in the presence of chromaffin granules. It turned out that the cages that confined granules rapidly disappeared when gelsolin was added in the presence of free Ca2+ ions. A semiquantitative analysis of dynamic light-scattering spectra permitted us to estimate the changes in the mobility (or the translational diffusion coefficient) of chromaffin granules in the actin network with its assembly and Ca(2+)-dependent disassembly by gelsolin. Based on the present results and some pieces of evidence in the literature, a model is proposed for biophysical situations before, during, and after an exocytotic event.

Actins↗
PubMed1993

Elastic filaments in situ in cardiac muscle: deep-etch replica analysis in combination with selective removal of actin and myosin filaments.

To clarify the full picture of the connectin (titin) filament network in situ, we selectively removed actin and myosin filaments from cardiac muscle fibers by gelsolin and potassium acetate treatment, respectively, and observed the residual elastic filament network by deep-etch replica electron microscopy. In the A bands, elastic filaments of uniform diameter (6-7 nm) projecting from the M line ran parallel, and extended into the I bands. At the junction line in the I bands, which may correspond to the N2 line in skeletal muscle, individual elastic filaments branched into two or more thinner strands, which repeatedly joined and branched to reach the Z line. Considering that cardiac muscle lacks nebulin, it is very likely that these elastic filaments were composed predominantly of connectin molecules; indeed, anti-connectin monoclonal antibody specifically stained these elastic filaments. Further, striations of approximately 4 nm, characteristic of isolated connectin molecules, were also observed in the elastic filaments. Taking recent analyses of the structure of isolated connectin molecules into consideration, we concluded that individual connectin molecules stretched between the M and Z lines and that each elastic filament consisted of laterally-associated connectin molecules. Close comparison of these images with the replica images of intact and S1-decorated sarcomeres led us to conclude that, in intact sarcomeres, the elastic filaments were laterally associated with myosin and actin filaments in the A and I bands, respectively. Interestingly, it was shown that the elastic property of connectin filaments was not restricted by their lateral association with actin filaments in intact sarcomeres. Finally, we have proposed a new structural model of the cardiac muscle sarcomere that includes connectin filaments.

Actin Cytoskeleton↗
PubMed1993

Time-resolved electron microscopic analysis of the behavior of myosin heads on actin filaments after photolysis of caged ATP.

The interaction between myosin subfragment 1 (S1) and actin filaments after the photolysis of P3-1-(2-nitrophenyl)ethyl ester of ATP (caged ATP) was analyzed with a newly developed freezing system using liquid helium. Actin and S1 (100 microM each) formed a ropelike double-helix characteristic of rigor in the presence of 5 mM caged ATP at room temperature. At 15 ms after photolysis, the ropelike double helix was partially disintegrated. The number of S1 attached to actin filaments gradually decreased up to 35 ms after photolysis, and no more changes were detected from 35 to 200 ms. After depletion of ATP, the ropelike double helix was reformed. Taking recent analyses of actomyosin kinetics into consideration, we concluded that most S1 observed on actin filaments at 35-200 ms are so called "weakly bound S1" (S1.ATP or S1.ADP.Pi) and that the weakly bound S1 under a rapid association-dissociation equilibrium with actin filaments can be captured by electron microscopy by means of our newly developed freezing system. This enabled us to directly compare the conformation of weakly and strongly bound S1. Within the resolution of deep-etch replica technique, there were no significant conformational differences between weakly and strongly bound S1, and neither types of S1 showed any positive cooperativity in their binding to actin filaments. Close comparison revealed that the weakly and strongly bound S1 have different angles of attachment to actin filaments. As compared to strongly bound S1, weakly bound S1 showed a significantly broader distribution of attachment angles. These results are discussed with special reference to the molecular mechanism of acto-myosin interaction in the presence of ATP.

Actin Cytoskeleton↗
PubMed1993

[Correlation between DNA content and amplification of oncogenes (c-myc, L-myc, c-erbB-2) and correlation with prognosis in 143 cases of resected lung cancer].

DNA content analysis using flow cytometry and amplification of c-myc, L-myc, and c-erbB-2 oncogenes in 143 cases of resected lung cancer were analyzed using the same specimen, and we examined the correlation with prognosis of DNA content and amplification of oncogenes. There were 54 DNA diploid cases (38%), 81 DNA aneuploid cases (57%) and 8 DNA multiploid cases. Analysis of oncogene amplification revealed 22 cases of c-myc, 4 cases of L-myc, and 22 cases of c-erbB-2. In curatively resected cases, the 5-year survival rate was 65% in 31 DNA diploid cases, and 36% in 40 DNA aneuploid cases. There was a statistically significant difference between the two groups (p < 0.02). However, in non-curatively resected cases, the 5-year survival rate was 11% in 23 DNA diploid cases, and 33% in 49 DNA aneuploid cases. There were no statistically significant differences among these groups. The correlation between DNA content and amplification of oncogenes was as follows. In DNA diploid cases, there were 4 cases of c-myc, and 6 cases of c-erbB-2. In DNA aneuploid cases, there were 15 cases of c-myc, 4 cases of L-myc, and 15 cases of c-erbB-2. In DNA multiploid cases, there were 3 cases of c-myc, and 1 cases of c-erbB-2. Amplification of oncogenes was seen more frequently in DNA aneuploid and multiploid cases than in DNA diploid cases. In 71 curative resected cases, the 5-year survival rate for amplified cases of c-myc (10 cases) was 0%, and that of cases with no amplification was 61% (no statistically significant difference). The 5-year survival rate for amplified cases of c-erbB-2 (10 cases) was 40%, against 52% for cases with no amplification. DNA content analysis using flow cytometry was more convenient than analysis of amplification of oncogenes, and reflects the prognosis of resected lung cancer better than oncogenes. There was no relation between DNA content and gene amplification.

DNA, Neoplasm↗
PubMed1992

The role of mediastinoscopic biopsy in preoperative assessment of lung cancer.

Between 1970 and 1989, mediastinoscopy and thoracotomy were performed on 619 patients admitted to our clinic with lung cancer. When mediastinoscopy was analyzed by lymph node location, the highest sensitivity (95.7%) was for the left paratracheal nodes and the lowest (64.0%) was for nodes at the bifurcation (p < 0.01). The 5-year survivals according to the results of mediastinoscopy were 47% for negative results, 14% for false-negative results, and 6% for positive results. The 5-year survival rate however, was significantly higher (28%) in patients (n = 13) with positive mediastinoscopic findings who underwent complete resection of the primary tumor and all involved nodes than in patients (n = 78) who underwent incomplete resection (p < 0.01). These data support our opinion that patients with positive mediastinoscopic results should not always be excluded from treatment by thoracotomy. The role of mediastinoscopy is not to select patients for thoracotomy but to evaluate lung cancer at the pretreatment stage.

Biopsy↗
PubMed1992

[Diagnosis, incidence, clinicopathology and surgical treatment of acquired tracheobronchomalacia].

We report our recent findings on the diagnosis, incidence, appearance of the trachea at autopsy, and surgical treatment of acquired trachobronchomalacia. In the diagnosis of this disease, continuous dynamic CT scanning (1 slice/0.6 second) from inspiration to deep expiration was better than X-ray cine recording. It is difficult to observe the most collapsed airway on coughing using bronchofiberscopic recording, because its duration time was estimated to be 1/10 second by X-ray cine recording, whereas the findings at one instant were easily recorded by video-bronchoscopy on coughing. The degree of severity of this disease was classified into three groups. 1st degree 0-50% narrowing of airway caliber, 2nd degree 50-75%; and 3rd degree 75-100%. Severity of 2nd or 3rd degree was present in 542 (12.7%) of 4283 patients suffering from chest disease who underwent bronchoscopy. Seventy-two percent of patients were aged 50 to 80 years. Third degree tracheomalacia was found in 131 patients with an incidence of 3.1%, which increased from 2.2% at 50 years to 6.2% at 80 years. The autopsy findings of the trachea consisted of fragmentation of the tracheal cartilage and extension of the membranous portion. The pathogenesis may be varied with different types of collapse of the airway, including the saber sheath type and the crescent type. Surgical treatment consists of prevention of the airway collapse by wrapping the airway with Marlex mesh and bonding Lyodura (lyophilized dura mata) with fibrin glue, which is more reliable than the span plasty proposed by Nissen.

Adolescent↗
PubMed1992

[Surgical treatment of lung cancer with mediastinoscopic positive lymph nodes].

There are no definite criteria for the indication of surgery in lung cancer with mediastinal lymph node involvement. During the past 20 years, 100 patients (76 patients with adenocarcinoma and 24 patients with squamous cell carcinoma) have undergone thoracotomy for lung cancer with mediastinoscopic positive lymph nodes at our hospital. Of these, relatively curative resection was performed on 13 patients. The 5-year survival rate in these 13 patients was 28%, which was significantly higher than the 0% in 42 patients with relatively non-curative resection and the 0% in 26 patients with absolutely non-curative resection. The 5-year survival rate was 9% in both T1 (n = 14) patients and T2 (n = 37) patients. No T3 (n = 21) and T4 (n = 9) patients survived 3 years. The 5 year survival rate in patients with squamous cell carcinoma was 12% and that in patients with adenocarcinoma was 0%, but there was no significant difference. The survival rates of T1 and T2 patients were significantly higher than that of T3 patients (p less than 0.02 and p less than 0.005) respectively. Contralateral mediastinal lymph node metastasis (N3) was observed significantly more frequently in patients with adenocarcinoma (38%) than in those with squamous cell carcinoma (13%), but there was no significant difference in the survival rate. In N2 patients, the survival rate was compared between those with mediastinal nodal involvement of an early stage (N2-1) and those with lymph node metastasis of more advanced stage (N2-2) according to the lobe bearing the primary cancer.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenocarcinoma↗