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Biomedical subjects

Y Tagawa

Publications and source records attributed to Y Tagawa.

At least 163 records · Page 9Linked to original sources

Mouse brain opioid receptor identification by direct ultraviolet photoaffinity labeling.

[3H]Morphine, PL-017[prolyl-3,4-3H,D-prolyl,3,4-3H] ([3H]PL-017) and enkephalin-(2-D-penicillamine,5-D-penicillamine)[tyrosyl-2,6-3H] ([3H]DPDPE) were directly cross-linked to mouse brain opiate receptors by an ultraviolet (254 nm) irradiation procedure. [3H]Morphine preferentially and specifically labeled a 58 kDa protein. The labeling of this protein was suppressed by the addition of excess naloxone. Dithiothreitol reduced the irreversible binding of [3H]morphine and [3H]PL-017 to the receptor protein. In the acid hydrolysate of [3H]DPDPE-labeled opiate receptors, dityrosine, was detected. These results suggest that the [3H]tyrosine residue of [3H]DPDPE covalently bound the tyrosine residue of delta-opioid receptors. The direct UV-photoaffinity labeling method using commercially available radioactive opiates described here should be a useful tool for characterization and purification of the opiate receptors.

Affinity Labels↗

Esophagectomy for metastatic carcinoma of the esophagus from lung cancer.

A patient with metastatic carcinoma of the esophagus from lung cancer is reported. The patient was a 54-year-old woman who underwent a left lower lobectomy for lung cancer 5 years previously. The authors performed a thoracic esophagectomy, dissection of mediastinal lymph nodes, and reconstruction of the esophagus; the surgery was followed by chemotherapy. Because the histologic pattern of the esophageal tumor was similar to that of lung cancer and mucosal involvement was not seen, the esophageal tumor was interpreted to be a metastasis from lung cancer. The patient is well without recurrence of disease 23 months after operation. This is the first report of a successful esophagectomy for metastatic carcinoma of the esophagus from lung cancer.

Adenocarcinoma↗

Comparison of pepsin and trypsin digestion on paraffin-embedded tissue preparation for DNA flow cytometry.

Preparation with enzymes plays an important part in obtaining good results in cellular DNA content measurements from paraffin-embedded tissue using flow cytometry. Therefore, we have compared two groups of DNA histograms obtained with pepsin (Hedley method) and trypsin digestion (Schutte method). Samples of five tumor types were compared: eight lung, seven breast, five thyroid, seven stomach, and six colon cancer cases. All samples were stained with propidium iodide (PI). The incidence of DNA aneuploidy determined by the Schutte method indicated a higher ratio in four tumor types than that determined by the Hedley method. Concerning the coefficient of variation (CV) used to estimate the quality of the DNA histogram, the diploid G1 peak determined by the Schutte method produced a smaller CV in five tumor types than did the Hedley method. This Schutte method had especially good results in lung and thyroid cancer. Furthermore, the amount of debris (background) determined by the Schutte method indicated a decrease in four tumor types compared with the Hedley method. In cell-cycle analysis of ten DNA diploid cases, DNA histograms by the Hedley method showed an increased S-phase fraction due to an overlapping of debris and aggregated cells. These results indicate that preparation by trypsin digestion is a method superior to pepsin digestion when the sample from paraffin-embedded tissue is stained with PI for the purpose of DNA content measurement.

Breast Neoplasms↗

The RNA content of alveolar macrophages in canine lung allografts.

To evaluate the utility of RNA synthesis by alveolar macrophages (AM) in lung allografts as a marker of acute rejection, we performed canine left lung allotransplantation and measured the AM RNA content during immunosuppressive therapy with azathioprine (AZ), cyclosporine A (CsA), or CsA combined with AZ and prednisolone (Pr). We determined the RNA content by determining the RNA index (RI) (peak channel number of AM/peak RNA channel number of peripheral blood lymphocytes) by flow cytometry. The dogs were classified into one of five treatment groups: Group 1 (n = 18), control (no surgery) dogs; Group 2 (n = 11), allotransplanted dogs without pre- or postoperative immunosuppressive therapy; Group 3 (n = 14), received AZ at dose of 5 mg/kg/day orally; Group 4 (n = 17), received CsA at dose of 20 mg/kg/day orally; Group 5 (n = 15), CsA combined with AZ (5 mg/kg/day) and Pr (0.5 mg/kg/day). The RI of normal lungs was 2.14 +/- 0.20 (mean +/- SD) (n = 18). The RI of rejected lungs was significantly higher than the RI of normal lungs in Group 2 (2.51 +/- 0.21, n = 8, P < 0.05) or Group 3 (2.76 +/- 0.36, n = 7, P < 0.05), but did not differ significantly from that in Group 4 (2.19 +/- 0.20, n = 7) or Group 5 (2.05 +/- 0.45, n = 8). The RI of rejected lungs was higher than the RI of nonrejected lungs in Group 2 (vs 2.18 +/- 0.12, n = 3, P < 0.05) or in Group 3 (vs 2.18 +/- 0.22, n = 7, P < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Combined transection of the trachea and esophagus following cervical blunt trauma.

The successful management of a patient with combined transection of the cervical trachea and esophagus following blunt trauma is reported. This type of injury was rare in the past, but is gradually increasing at the present time because of the increased use of motorcycles. Immediate primary closure of the transected trachea and esophagus offers the best chance for a good result.

Accidents, Traffic↗

Purification and partial characterization of the major outer membrane protein of Haemophilus somnus.

We purified the major outer membrane protein (MOMP), which is the most abundant OMP (with an apparent molecular mass of 40 kDa), from Haemophilus somnus strain 8025. The method involves solubilization of the MOMP with Zwittergent 3-14 and further purification accomplished by ion-exchange and molecular-sieve chromatographies. The amino-terminal sequence of the MOMP showed considerable similarity to those of porin proteins from other gram-negative bacteria. The MOMP of H. somnus is immunogenic to rabbits and calves. Hyperimmune sera from rabbits and calves reacted with both the MOMP and lipopolysaccharides in enzyme-linked immunosorbent assay (ELISA) and immunoblot analysis. The rabbit antiserum to the MOMP was cross-reactive with whole-cell preparations from strains 8025, D1238, NT2301, and 540 at a band with a molecular mass of 40 kDa in immunoblot analysis, although the reactivity of the rabbit antiserum with strain 540 was lower than those with the other strains tested. Two murine monoclonal antibodies (MAbs) to the MOMP were developed. ELISA with the OMP fractions as the antigens showed that one MAb was cross-reactive with the four strains but that the other MAb was reactive with the three strains other than strain 540. These results indicate that the MOMP of H. somnus possesses at least two antigenic determinants and that the MOMP of strain 540 is antigenically different from those of the other strains. The antigenic heterogeneity of the H. somnus MOMP has implications regarding the development of a serotyping system with MAbs that is based on the MOMP epitopes.

Amino Acid Sequence↗

Characterization of an immunoreactive 17.5-kilodalton outer membrane protein of Haemophilus somnus by using a monoclonal antibody.

A single outer membrane protein (OMP) of Haemophilus somnus, with an apparent molecular mass of 17.5 kDa, was identified in the sodium dodecyl sulfate (SDS)-insoluble fraction after extraction with 1% SDS-0.5 M NaCl-0.1% beta-mercaptoethanol. A hybridoma derived from mice immunized with H. somnus OMP fractions produced a monoclonal antibody (MAb), designated 20-3-5, that bound to the 17.5-kDa OMP of H. somnus. The MAb 20-3-5 epitope was present on 45 of 45 strains of H. somnus tested. MAb 20-3-5 cross-reacted with Haemophilus agni, Histophilus ovis, and Haemophilus haemoglobinophilus but not with 13 other species and subspecies of gram-negative bacteria. Immunoelectron-microscopic and antibody absorption studies revealed that the MAb 20-3-5 epitope is exposed on the surface of bacteria. In an immunoblot analysis, convalescent-phase sera obtained from calves with experimental H. somnus pneumonia contained antibodies to the 17.5-kDa OMP of H. somnus. Future studies will be directed toward examining the role of the 17.5-kDa OMP in immunity to H. somnus infections.

Animals↗

Characterization of a heat-modifiable outer membrane protein of Haemophilus somnus.

In immunoblot analysis, a murine monoclonal antibody (MAb), 27-1, which was produced to an outer membrane protein (OMP) of Haemophilus somnus, showed that a major OMP is heat modifiable, having a molecular mass of 28 kDa when the N-lauroylsarcosine-insoluble OMP preparation was solubilized at 60 degrees C and a mass of 37 kDa when the OMP preparation was solubilized at 100 degrees C. The heat-modifiable OMP reacted intensely with convalescent sera obtained from calves with experimental H. somnus pneumonia in immunoblot analysis. Immunoelectron microscopic and antibody absorption studies revealed that the MAb 27-1 epitope was not surface exposed on the intact bacterium. However, a decrease in antibody reactivity to the heat-modifiable OMP in immunoblot analysis after absorption of convalescent serum with intact bacterial cells of H. somnus suggests that a surface-exposed portion of the heat-modifiable OMP is expressed on the intact bacterium. MAb 27-1 reacted with 45 of 45 strains of H. somnus tested in immunoblot analysis. The apparent molecular mass of the antigen varied among strains, and five reactivity patterns demonstrated by MAb 27-1 were observed. MAb 27-1 also reacted with six species in the family Pasteurellaceae, Escherichia coli, and Salmonella dublin, but not with the other eight species of gram-negative bacteria. The heat-modifiable OMP of H. somnus showed immunological cross-reactivity with the OmpA protein of E. coli K-12 and significant N-terminal amino acid sequence homology with the OmpA proteins of gram-negative bacteria. We conclude that a major, 37-kDa heat-modifiable OMP of H. somnus, which elicits an antibody response in H. somnus-infected animals, is a common antigen among H. somnus strains tested and is structurally related to the OmpA protein of E. coli.

Amino Acid Sequence↗

Antigenic analysis of the major outer membrane protein of Haemophilus somnus with monoclonal antibodies.

The major outer membrane protein of Haemophilus somnus possesses at least five distinct epitopes. Three surface-exposed epitopes on the major outer membrane protein include a conserved epitope with potential for development of a vaccine and a diagnostic test and two variable epitopes responsible for antigenic differences among strains; the remaining two epitopes are well preserved among strains but not exposed on the cell surface.

Antibodies, Bacterial↗

Modifying effects of chemicals on the development of liver preneoplastic placental glutathione S-transferase positive foci in analbuminemic and Sprague-Dawley rats.

Nagase analbuminemic rats (NARs) were compared to the Sprague-Dawley (SD) stock in a medium-term assay system for hepatocarcinogenesis regarding their susceptibilities to the influence of chemicals on the development of glutathione S-transferase, placental form, positive (GST-P+) foci. Two weeks after initiation with diethylnitrosamine (DEN), the animals were exposed alternatively to 0.06% 3'-methyl-4-dimethyl-aminoazobenzene (3'-Me-DAB), 50 ppm DEN, 0.25% ethionine, 1% clofibrate, and 1% butylated hydroxyanisole (BHA) for a 6-wk period. Adequate controls included groups only initiated with DEN or treated with each test compound alone. For evaluation of the modifying potential of the chemicals, indices were generated by using the mean values obtained for number and area of GST-P+ foci after each treatment. Comparison between these indices suggests that SD rats were relatively more sensitive than NARs to the modifying effects of complete carcinogens (3'-Me-DAB and DEN). The strains were similarly-susceptible to the promoting influence of ethionine, a nongenotoxic carcinogen. The inhibitory influence of BHA was more intense in NARs, whereas in both strains clofibrate was associated to similarly reduced values for number and area of GST-P+ foci. The degree of susceptibility of each strain to the modifying influence of chemicals on foci development depended on the chemical agent investigated.

Acetylglucosaminidase↗

[Lung preserving operation for bronchogenic carcinoma--segmentectomy and bronchoplastic lobectomy].

The results of segmentectomy and bronchoplastic lobectomy for bronchogenic carcinoma were reported. Until December 1992, 920 patients underwent lung resection for primary lung cancer at our institution. Among them, 70 patients (7.6%) had segmentectomy and 149 (16.2%) had bronchoplastic lobectomy. In cases with segmentectomy, adenocarcinoma (57.1%) and Stage I disease (77.1%) were dominant, and in cases with bronchoplasty, m squamous cell carcinoma (69.8%) and Stage IIIA (45.0%) were frequent. There were no operative deaths and one hospital death (1.4%) in patients with segmentectomy, and 8 operative deaths (5.4%) and 9 hospital deaths (6.0%) in those with bronchoplasty. Five-year survival rates of the patients with segmentectomy were 49.6% in total and 64.6% in those with Stage I disease. Five-year survival rates of those with bronchoplasty were 33.9% in total, 51.1% in those with Stage I, 29.1% in those with Stage IIIA and 39.0% in those with squamous cell carcinoma. Respiratory function was well preserved in patients with segmentectomy and bronchoplastic lobectomy. Lung-preserving operations such as segmentectomy and bronchoplastic lobectomy are meaningful as a curative procedure for primary lung cancer for selected patients.

Adenocarcinoma↗

[Relationship between the Ki-67 nuclear antigen content and cell-cycle perturbation on WiDr cells treated with 5-FU].

The profile of Ki-67 nuclear antigen content according to the change in the cell-cycle of WiDr cells affected by 5-fluorouracil (5-FU) was studied by flow cytometry. The cases who received 10 micrograms/ml for 1 hr and showed reproliferation in the growth curve presented significant accumulation at G2M phases. However, significant increase of Ki-67 antigen content was not observed in G2M phases. Meanwhile, the cases who continuously received 10 micrograms/ml for 72 hr and showed the suppression of cellular proliferation without reproliferation presented gradual accumulation into S phase and Ki-67 antigen content was found to be significantly increased. This increase occurred in G1 phase, and the cells with Ki-67 antigen content of the largest increase in G1 phase shifted to S phase. It indicates a possibility that Ki-67 nuclear antigen content is destined to be controlled specifically in G1 phase of the cell-cycle. 5-FU seems to damage this regulation system and, as a result, increase Ki-67 nuclear antigen.

Cell Cycle↗

[Vanishing tumor thrombus in IVC after oral administration of UFT and TAE in a patient with HCC; a case report].

A 74-year-old man was diagnosed as hepatocellular carcinoma (HCC) in the area of S6-7 with tumor thrombus continuously developed from hepatic vein to inferior vena cava (IVC). AFP value remained high despite two transcatheter arterial embolizations with adriamycin, mitomycin and Lipiodol (Lp-TAE). The AFP value gradually decreased two weeks after oral administration of UFT. Nine months after, the AFP level was 26.4 ng/ml and the tumor thrombus in the IVC disappeared. The size of the main tumor did not change. This case suggests Lp-TAE and UFT are useful for suppressing the growth of the main tumor and to extinguish the tumor thrombus.

Aged↗

[Comparison of typical and atypical bronchoplasty for bronchogenic carcinoma].

One-hundred and twenty-eight patients who underwent bronchoplasty for bronchogenic carcinoma from 1969 to 1990 were retrospectively analyzed. Ninety-six patients had upper sleeve lobectomy or upper lobectomy with wedge bronchoplasty (typical procedure) and 32 underwent bronchoplasty with lobectomy other than upper lobes (atypical bronchoplasty). The two groups were statistically similar in preoperative characteristics including sex, age, cell types and stages. In typical group, right upper sleeve lobectomy was performed in 48 patients, left upper sleeve lobectomy in 27, right upper lobectomy with wedge bronchoplasty in 11, left upper lobectomy with wedge bronchoplasty in 7 and sleeve bronchoplasty alone in 2. In atypical group, right middle and lower lobectomy with sleeve bronchoplasty in 10 patients, left lower sleeve lobectomy in 5, right lower lobectomy with wedge bronchoplasty in 4 and others in 13. Postoperative mortality was 5.2% in the typical group and 9.4% in the atypical group. Postoperative respiratory complications occurred in 16 patients (50%) in the patients with the atypical group and in 33 (34.7%) in those with typical group. Three years and 5 years survival rates in the typical group were 46 and 40% and those in the atypical group, 27 and 18%. However, there was no statistical difference in survival between the two groups. In conclusion, atypical bronchoplasty is useful for preserving the lung parenchyma, but, careful perioperative management and surgical technique of bronchoplasty are mandatory.

Adult↗

[Detection of chromosomal aberration using fluorescence in situ hybridization in DNA diploid colorectal carcinomas].

Numerical aberrations of chromosomes can be detected by fluorescence in situ hybridization (FISH), using chromosome-specific probes. It is possible to observe this in solid tumors from which it is very difficult to obtain metaphase nuclei. The present study employed surgical specimens from 15 cases of colorectal carcinoma, all of which showed DNA diploidy. In the same samples, we analyzed chromosomal numerical aberration by FISH according to the method of Pinkel et al. Biotinylated DNA probes specific to chromosome #7, #11 and #17, were used. The hybridization spots were observed by fluorescent microscopy. As a result, the numerical aberrations of chromosomes detected by FISH were found in DNA diploid cases by FCM. They were trisomy and monosomy. These results indicate that FISH is useful to detect the chromosomal aberrations of DNA diploid cases.

Chromosome Aberrations↗

Extraordinarily stable mini-hairpins: electrophoretical and thermal properties of the various sequence variants of d(GCGAAAGC) and their effect on DNA sequencing.

A small DNA fragment having a characteristic sequence d(GCGAAAGC) has been shown to form an extraordinarily stable mini-hairpin structure and to have an unusually rapid mobility in polyacrylamide gel electrophoresis, even when containing 7M urea. Here, we have studied the stability of the various sequence variants of d(GCGAAAGC) and the corresponding RNA fragments. Many such sequence variants form stable mini-hairpins in a similar manner to the d(GCGAAAGC) sequence. The RNA fragment, r(GCGAAAGC) also forms a mini-hairpin structure with less stability. The DNA mini-hairpins with GAAA or GAA loop are much more stable than DNA and RNA mini-hairpins with other loop sequence so far as has been examined. The stability difference between DNA and RNA mini-hairpins may be deduced to the stem structures formed by DNA (B form) and RNA (A form). The stable hairpins consisting of the GCGAAAGC sequence cause strong band compression on the sequencing gel. This phenomenon should be carefully considered in DNA sequencing.

Base Sequence↗

Lack of effects of post-initiation cholesterol on 3,2'-dimethyl-4-aminobiphenyl-induced prostate carcinogenesis.

The effects of cholesterol on 3,2'-dimethyl-4-aminobiphenyl (DMAB)-induced rat prostate carcinogenesis were investigated in male F344 rats. Animals were given 10 subcutaneous (s.c.) injections of 50 mg/kg body weight of DMAB at 2-week intervals, each time 2 days after transfer from 1 week on 0.75 ppm of an ethinyl estradiol (EE)-supplemented diet to a basal diet. From week 20, the animals received treatment with cholesterol (2 or 1%, in the diet), saponin (1 or 0.2 mg/rat/week, s.c.), or clofibrate (0.3%, in the diet) for 40 weeks. Although serum cholesterol levels were significantly increased by administration of cholesterol itself, and were decreased by clofibrate, the incidences of atypical hyperplasias and carcinomas of the ventral prostate were very similar with all treatments. Thus, under the present conditions, high or low cholesteremia did not affect rat prostate lesion development.

Aminobiphenyl Compounds↗

Upper airway obstruction from a benign goiter.

Acute upper tracheal obstruction from a huge benign goiter developed in an 80-year-old woman. Computed tomography showed severe stenosis of the cervical trachea by extrinsic compression. A left hemithyroidectomy was performed and the tracheal lumen returned to normal size. However, the tracheal wall was malacic due to the prolonged compression by the goiter and a silastic T-tube insertion was necessary for a two month period. This type of tracheal obstruction is rare. The importance of evaluating the upper trachea in patients with prolonged benign goiter is stressed and the diagnosis as well as the management of this type of complication are discussed.

Aged↗