cDNA cloning of nine S alleles and establishment of a PCR-RFLP system for genotyping European pear cultivars.

Nine full-length cDNAs of S ribonucleases (S-RNases) were cloned from stylar RNA of European pear cultivars by RT-PCR and 3' and 5' RACE. Comparison of the nucleotide sequences between the nine S-RNases cloned and 13 putative S alleles previously amplified by genomic PCRs revealed that seven corresponded to Sa, Sb, Sd, Se, Sh, Sk and Sl alleles, and the other two were new S alleles (designated as Sq and Sr alleles). Genomic PCR with a set of a8FTQQYQa9 and a8EP-anti-IIWPNVa9 primers was used to amplify nine S alleles; 1,414 bp (Sl), ca. 1.3 kb (Sk and Sq), 998 bp (Se), 440 bp (Sb) and ca. 350 bp (Sa, Sd, Sh and Sr). Among these, S alleles of similar size were discriminated by digestion with BaeI, BglII, BssHII, HindIII, EcoO109I and SphI. The PCR amplification of S allele following digestion with the restriction enzymes provided a PCR-RFLP system for rapid S-genotyping European pear cultivars harboring nine S alleles. The PCR-RFLP system assigned a total of 63 European pear cultivars to 25 genotypes. Among these, 14 genotypes were shared by two or more cultivars, which were cross-incompatible. These results suggested that the genes cloned represented the S-RNases from European pear, and that there were many cross-incompatible combinations among European pear varieties.

[True pulmonary carcinosarcoma suspected as a metastatic lung tumor].

This patient, a 53-year-old man, has underwent operation on the diagnosis of esophageal cancer 2 years ago. An abnormal shadow was detected in the left lung field and he was admitted to our hospital for further examination. On the suspicion of metastatic lung tumor by transbronchial brushing cytology, partial resection of the left lower lobe was performed. Histologically the tumor was of carcinomatous (squamous cell carcinoma and adenocarcinoma) and sarcomatous (fibrosarcoma and chondrosarcoma) elements, so the patient was diagnosed as "true" pulmonary carcinosarcoma. True pulmonary carcinosarcoma is one of the very rarest neoplasms of the lung.

Inheritance of the Md-ACS1 gene and its relationship to fruit softening in apple ( Malus x domestica Borkh.).

The 1-aminocyclopropane-1-carboxylic acid synthase (ACS) gene is a member of the ACS gene family that is involved in apple ( Malus x domestica Borkh.) fruit ripening. Presence of an allele ( Md-ACS1-2) of this gene is associated with low internal ethylene concentration in some apple cultivars. In this study, inheritance of Md-ACS1 was determined for 50 apple cultivars/advanced selections and 101 F(1) seedlings from five populations. Following this, the softening pattern of apples stored at 20 degrees C for up to 40 days was examined using 35 fruiting cultivars/selections of defined Md-ACS1 status. Md-ACS1 is inherited in a Mendelian fashion and was found to be linked to fruit softening. Maturity season of genotypes also significantly affected fruit softening. Late-season genotypes in the Md- ACS1-2/2 class had the slowest rate of softening, while early-season Md- ACS1-1/1 genotypes had the most rapid softening rate. The implications of these results are discussed in relation to parental selection and breeding for storage ability in apple.

Usefulness of enzyme-linked immunosorbent assay using recombinant desmogleins 1 and 3 for serodiagnosis of pemphigus.

Pemphigus is an autoimmune blistering disease with two major subtypes, pemphigus vulgaris (PV) and pemphigus foliaceus (PF). Patients with pemphigus have circulating antidesmoglein (Dsg)1 and/or anti-Dsg3 IgG autoantibodies. We have previously developed enzyme-linked immunosorbent assays (ELISAs) using recombinant Dsg1 and Dsg3 expressed by baculovirus as a diagnostic tool for pemphigus. The purpose of this study was to evaluate the practical application of these ELISAs for clinical use with a large number of serum samples. We used 81 PV sera, 48 PF sera, 114 bullous pemphigoid (BP) sera, 124 collagen disease sera, nine sera of other non-pemphigus bullous diseases and 179 normal control sera. A cut-off value was determined by receiver-operating-characteristic plots. Forty-seven of 48 PF sera (97.9%) were positive in the Dsg1 ELISA and 79 of 81 PV sera (97.5%) were positive in the Dsg3 ELISA, while only two (1. 1%) and four (2.2%) of 179 normal sera were positive in Dsg1 and Dsg3 ELISAs, respectively. However, some disease control sera of BP and collagen diseases exceeded the cut-off value. Introduction of a grey zone helped to decrease the number of these false-positive sera. Furthermore, in three patients studied, the respective Dsg1 and Dsg3 ELISA scores showed parallel fluctuation with the disease activity along the time course. We conclude that Dsg1 and Dsg3 ELISAs provide a simple, sensitive and highly specific assay for the diagnosis of patients with PV and PF and that these ELISAs may be a valuable tool to monitor the disease activity. We also propose diagnostic criteria for pemphigus based on ELISA reactivity: if a serum is positive against Dsg3 it indicates a diagnosis of PV, regardless of reactivity against Dsg1; if a serum is negative for Dsg3 and positive for Dsg1, it indicates a diagnosis of PF.

[Pulmonary vein reconstruction for lung carcinoma].

Four cases of primary lung carcinoma (two squamous cell carcinomas and two adenocarcinomas) were performed right middle lobectomy combination with the reconstruction of upper pulmonary vein to remain the upper lobe. Lung carcinomas were in the right lobe and infiltrated to lower margin of upper pulmonary vein in all cases. After resection of the right middle lobe and an affected portion of upper pulmonary vein, the defect of the upper pulmonary vein was replaced with auto-pericardial graft in three cases, and the other one was closed by continuous suture of 5-0 plorene. Pathological classification of these four cases was stage IB in one patient, stage IIB in two and stage IV in one. All patients died from 6 to 53 months after operation (average: 31 months). Metastasis to distant organs was confirmed in all cases, so the prognosis of them was generally poor. Reconstruction of pulmonary vein may be feasible to avoid over resection of other lobes, because pulmonary function can be preserve as well as in the bronchoplasty.

[A case of medullary carcinoma of the thyroid gland: metachronous bilateral metastasis of the neck and mediastinal lymph nodes were treated successfully by staged resection].

This patient, a 52-year-old male, underwent subtotal thyroidectomy on the diagnosis of medullary carcinoma of the thyroid gland in 1980 and postoperative course was uneventful. Since November 1990 he had a persistent diarrhea for 6 months and was admitted to the hospital for the further examination on June 1991. The serum CEA and calcitonine level was very high and chest CT scan findings showed the swelling of right neck and mediastinal lymph nodes. Dissection of the lymph nodes was performed by anterior approach which was gained through a proximal median sternotomy extended into the anterior fourth intercostal space as well as to the base of the neck on the right side. On the pathological examination it was metastasis of medullary carcinoma of the thyroid gland. And 50 months later after second operation he had a persistent diarrhea once again. Left neck and mediastinal lymph node metastasis was detected by chest CT with high serum CEA and calcitonine level. Similarly resection was performed by the same anterior approach on the left side. Irrespective of the extended resection he was free of severe complication; he is still alive 10 months after the third operation without any evidence of recurrence and his current performance status is very good.

[Assessment of surgery for primary lung cancer with dissemination or malignant effusion of pleura (T4 advanced lung cancer)].

Twenty-four cases of primary lung cancer with dissemination or malignant effusion of pleura detected preoperatively or intraoperatively were surgically treated at our hospital. Mean survival time (MST) and two-year survival rate (2 YSR) were analyzed on their resected cases and non-resected cases with similar lesion. MST and 2 YSR of 19 cases with lobectomy, 5 cases with pneumonectomy including pleuropneumonectomy and 15 cases with no surgical procedure were 2.77 +/- 0.60 years, 53.4%, 1.51 +/- 0.50 years, 26.7% and 0.99 +/- 0.15 years, 6.7%. MST and 2 YSR of 13 cases with lymph node dissection under R 1 and 6 cases over R 2 on lobectomy group were 1.99 +/- 0.38 years, 37.5% and 5.66 +/- 1.71 years, 66.7%. These findings suggested that lobectomy with lymph node dissection of R 2 over may be a beneficial treatment of lung cancer with dissemination or malignant effusion of pleura.

[A case of lung adenocarcinoma: detection and resection of the metastatic lesion was managed effectively by serum CEA level].

This patient, a 53-year-old male, has had back pain and an abnormal shadow was detected in the right lung field on December 1989. He was admitted to the hospital for the further examination. On the diagnosis of lung cancer with high serum CEA level operation was performed on February 1990. As a results of pathological examination, histological type was adenocarcinoma and pathological stage was pT3N0M0 stage IIIA. After operation the serum CEA level was decreased immediately but it was gradually increased once again. And then 14 months later right adrenal metastasis was detected by abdominal CT with high serum CEA level and resection was performed. Similarly a solitary lymph node metastasis located in abdomen was detected and resected with high serum CEA level 28 months after second operation. In this case detection and resection of the metastatic lesion was managed effectively by serum CEA level. The patient had a good operative course and is alive 76 months after first operation without any evidence or recurrence.

Bronchoscopic findings of extramural lung cancer invading the subepithelium or submucosa.

Bronchoscopic findings from the main to segmental bronchi were compared with the histopathological findings in 185 resected cases of lung cancer, in order to determine which bronchoscopic features are associated with lung cancer invading the subepithelium or submucosa from beyond the bronchial wall. Carcinoma invaded the subepithelium or submucosa from beyond the bronchial wall in 43 cases (22.9%) out of the total of 185 cases. Bronchoscopic findings were evaluated in these 43 cases, and were summarized as follows: (1) The bronchoscopic findings in cases of subepithelial invasion consisted of vascular engorgement, bleeding, subepithelial tumor, and emphasized longitudinal relief; (2) irregularity of the mucosa was observed in cases of epithelial or muscular invasion; (3) indistinct bronchial cartilage was observed in cases of invasion proximal to the extramuscular layer; (4) accentuated irregular folds were observed in cases of invasion of the extramuscular or cartilage layers; and (5) edema and redness were not specific for malignancy. In addition, the occurrence of mediastinal lymph node metastasis was higher in cases of invasion to main or lobar bronchi. This result indicates that recognition of invasion of the subepithelium or submucosa of the central bronchus may be helpful in indicating the probability of mediastinal lymph node metastasis. Accordingly, there may be specific bronchoscopic findings which correlate with invasion of the bronchial subepithelium or submucosa. Accurate recognition of these findings may be useful in determining appropriate biopsy sites and may provide more information concerning selection of therapeutic strategy.

[A rare case of intrapulmonary sequestration in left upper lung field diagnosed before surgery].

This case is a 30-year-old male who was indicated abnormal shadow in the left upper lung field by chest X-ray film. The aortography was carried out because pulmonary sequestration was suspected by chest CT-scan etc. From the result, rare pulmonary sequestration in the left upper lung field was diagnosed because patterns with inflow of an abnormal artery from descending aorta into the left upper lung field and perfusion of it into the upper pulmonary vein were observed. The wedge resection was performed. The specimen was slightly hard on the whole, and many cartilages or multiple nodules were observed on the cut surface. Histopathologically, normal pulmonary structure was disappeared, and hyperplasia of the lymphatic nodule was remarkably observed in the interstitium, and cystic wall was covered by ciliated cylindrical epithelium.

[A case of hepatic hydrothorax treated by pleuro-venous shunt].

We reported a successful pleuro-venous shunt operation (Pl-V shunt) for a patient of liver cirrhosis with secondary hydrothorax. A 78-year-old female was admitted to our hospital because of severe dyspnea and palpitation. Chest x-ray film revealed right sided massive pleural effusion. Over several weeks the chest tube drainaged about 1,500 ml of transudative fluid per day. We performed Pl-V shunting and pleural effusion subsequently decreased in amount and dyspnea disappeared. This Pl-V shunting is thought to be useful for such a patient with massive pleural effusion which failed to respond to medical therapy.

[A successfully treated case of empyema with a large tracheal fistula after a radical operation of esophageal cancer by fixation and plombage with major pectoral muscle flap].

We reported a successfully treated case of empyema with a large tracheal fistula which had developed after a radical operation of esophageal cancer (reconstructed with stomach). This 59-year-old male was treated by the method of fixation and plombage with major pectoral muscle flap and thoracoplasty, because we could not use the omentum that were frequently used nowadays for closure of the fistula. The size of the tracheal fistula was a large as the main bronchus bronchoscopically. Postoperative care were the following, the endotracheal tube was inserted from the tracheal stoma to the left main bronchus and 9 days left hemi-ventilation was performed. Continuous suction was performed at the same time from the right main bronchus in order to prevent secretion and blood pour into the left lung. Bronchoscopical examination done 28 postoperative day, the small fistula remained the tip of the muscle flap. But 72 postoperative day, the surface of the fixed muscle flap was replaced by normal bronchial mucomembrane and tracheal fistula was obliterated. Major pectoral muscle could be used as local flap to obliterate empyema cavity associated with tracheal fistula. We believe that utilizing an muscle flap for those who had undergone abdominal operation like our case is a valuable method.

[An experience of laser therapy of lung cancer complicated with hemophilia A].

We applied an endoscopic Nd-YAG laser therapy for an early hilar lung cancer complicated with hemophilia A in a patient of 62-year-old male. Abnormality was pointed out by means of sputum cytology. But, no shadow of tumor was observed by chest x-ray films and computed tomogram. By a bronchoscopy, a mild irregular mucomembrane and stenosis without tumor were exhibited. By means of cytological diagnosis of an abrasive-washing specimen of that region, the presence of squamous cell carcinoma was proved. Also, he was diagnosed as factor VIII deficient hemophilia A. So we performed him a laser therapy in place of the operation and with an intrabronchial arterial infusion. After the laser treatment, the conditions progressed better without any recurrence but about one and a half years later, an intrapulmonary metastasis occurred. Then, radiation and chemotherapy have been applied and at present, the patient is in a good state although under the tumor-bearing condition.

[Usefulness of the percutaneous marking method with a coil (PMMC) for the localization of small pulmonary nodules].

We surgeons have always encountered difficulty in localizing small pulmonary nodules, measuring less than 5 mm in diameter, through palpation during operation. In order to solve such a problem, we devised a new method of localizing small pulmonary nodules by applying a technique of CT-guided needle biopsy; namely, metallic spring coil is implanted through a 16 G eraster needle, the tip of which has been placed near the lesion. Our method not only secures the resection of entire lesions, but reduces an amount of lung resection through a small incision (mini-thoracotomy) as well. Furthermore, this method is also thought to be useful for open lung biopsy of pulmonary nodules, measuring less than 1 cm in diameter, in saving time and reducing an amount of lung tissue collection. In case of cystic lesion which is thought to be difficult to palpate on operation, this method can also be useful for its resection.

[Resected cases of middle lobe primary cancer].

Of 200 lung cancer lesions resected in our hospital, there were 15 cases (7.5%) with middle lobe origin. The histological types were adenocarcinoma in 13 patients (4 patients with alveolar cell carcinoma), squamous cell carcinoma in one and large cell carcinoma in one. These patients were classified into two groups according to the type of operation they received and each group was evaluated. Group I (resection of the middle lobe) included 8 patients. Each one of Stage IIIB and Stage IV received the operation to improve their symptoms. The six patients of Stage I received only middle lobectomy as absolute curable cases. Group II (resection of the middle and lower lobes) included 7 patients, who had preoperative diagnosis of stage III. Two of them were postoperatively found to be cases of Stage I and Stage II. Although it was still short-term, the follow-up evaluation proved that these patients survived without local recurrence and distant metastasis, except for two with pleural dissemination and one with cerebral metastasis, who had received lobectomy as palliative operation. No difference was observed between the two groups receiving different types of operation.

[The use of an omental pedicle flap in the closure of a wide bronchial fistula after pneumonectomy: a case report].

A 52-years-old man with pulmonary hypofunction had a squamous cell lung carcinoma and underwent the right upper lobectomy. Bronchial fistula with lung abscess developed on the 6th post-operative day. So, the right middle and lower lobectomy (completion pneumonectomy) were done. But, bronchial fistula again appeared at the time of weaning from respirator. It increased in size to about 4.0 x 2.0 cm. Then, transposition of an omental pedicle flap for closure on a wide bronchial fistula, and muscular plombage and thoracoplasty for semifilling up a secondary empyema cavity were performed. On bronchoscopy performed 14 days after operation, the fistula was completely closed, and the transposed omentum did not project into the trachea. We accomplished our first aim to close the wide bronchial fistula with omentum. However, he died 59 days after the 3rd operation because we failed to control infection of the remaining empyema cavity of about 100 ml. The management of an empyema cavity remains to be a difficult therapeutic problem.