Agrobacterium rhizogenes lacZ-rolC gene expression in Escherichia coli: detection of the product in transgenic plants using RolC-specific antibodies.

The rolC sequence of the Agrobacterium rhizogenes Ri plasmid was fused in-frame to the 3' end of the lacZ gene in plasmid pEX3. The fusion protein RolC-beta-galactosidase was accumulated as insoluble inclusion bodies in Escherichia coli. Antibodies were raised in rabbits against the fusion protein. After affinity purification, RolC-specific antibodies were found to react with a 22-kDa polypeptide prepared from roots of transgenic tobacco plants possessing a rolC gene. The result of differential centrifugation suggested that RolC is present in the soluble fraction of transformed cells.

Spontaneous antibody-secreting cells in the stomach of gastric cancer patients.

The gastric mucosa has been regarded as an active site of humoral immunity since the discovery of Helicobacter pylori. The present study was conducted to determine the in vivo activity of gastric B cells in 53 gastric cancer patients. B-cell activity was measured by protein-A plaque assay, in which IgA-, IgM-, and IgG-plaque-forming cells (PFC) were counted. The number of PFC was associated with the stage of cancer, but the response of lymphocytes in a non-tumorous area (NML) and tumor-infiltrating lymphocytes (TIL) differed. PFC in both sites were decreased compared to n0 cancer in n1 lymph node metastasis-positive cancer, while only NML showed raised PFC in n2 + (P < 0.05, vs TIL). Cancer cells penetrating the submucosa caused the PFC of TIL (but not of NML) to decrease. Invasion of the intratumor capillary (V) or lymphatic (Ly) vessels also caused PFC to change, showing differences of Ig class; there was a decrease of PFC in V2 (IgG- and IgM-PFC) and in Ly2 (all Ig-PFC). IgA-PFC in Ly1 differed in TIL (decrease of PFC) and NML (increase). PFC also differed in TIL and NML in cancer cells, as follows: TIL < NML in tubular and poorly differentiated adenocarcinoma and TIL > NML in papillary and signet ring cell adenocarcinoma. Changes in lymph node (LNL) and blood lymphocytes were similar to those in gastric PFC whose IgA value was 10 times as much as that of LNL. The 5-year survival rate was significantly better in patients with lower rather than higher PFC such as 89% vs 68%. Gastric B cells thus appear to be active and to reflect gastric mucosal immunity.

Chemical modification of streptovaricin C I. 19-O-substituted damavaricin C.

Damavaricin C, a degradative derivative of streptovaricin C, has reduced antibiotic activity relative to streptovaricin C. It has, however, a new phenolic hydroxyl group at the C-19 position of the naphthoquinone ring on which various groups can be substituted through an ether linkage. A series of 19-O-substituted derivatives of damavaricin C has been synthesized. The preparation of these derivatives, their in vitro antibacterial activities, in vitro inhibition of E. coli RNA polymerase, and lethal activity on the membrane mutants of E. coli are reported. It is believed that the original biological activity of damavaricin C is retained and that introduction of the functional groups at the C-19 position has increased the membrane diffusibility of the molecule.

Capsimycin, a new antibiotic. I. Production, isolation and properties.

Capsimycin is a new antifungal antibiotic produced by a strain of Streptomyces sp. C 49--87. The active substance in the fermented broth was isolated by solvent extraction followed by silica gel column chromatography. The antibiotic melts at 186 degrees C (decomp.) and has a molecular formula C30H40N2O6. It exhibits marked inhibitory activity against Phytophthora capsici (Leaf blight disease of cucumber) and Pythium debaryanum (Damping-off disease of cucumber).

A case of breast cancer diagnosed by inguinal lymph node metastasis.

We describe a case of a 58-year-old woman with right inguinal lymph node swelling and a T1 tumor in the right breast. She was referred with an 18-month history of the former complaint and a six-month history of the latter. Excisional biopsy of the inguinal lymph node revealed breast cancer metastasis. Radiographical examination showed no metastases to the lungs, liver or bone. Modified radical mastectomy was performed. Histological examination revealed solid tubular carcinoma, PT2, PM (axillary lymph node metastases 4/16), stage IV. Estrogen and progesterone receptors were negative. Three cycles of postoperative cyclophosphamide, adriamycin and 5-fluorouracil (CAF) chemotherapy were given, and the right inguinal area was irradiated with 40 Gy. The patient complained of swelling in both legs three years after surgery. Computed tomography revealed marked lymph node swellings in the pelvic cavity. She died six months later. Inguinal lymph node metastasis from breast cancer is very rare, although distant lymph node metastasis in the cervix occurs frequently. This case should help clarify how breast cancer metastasizes to distant lymph nodes.

[A case of malignant hemangioendothelioma of the lung with rapid growth of pulmonary metastasis].

Malignant hemangioendothelioma of the lung is very rare and difficult to diagnose. The growth is rapid and the prognosis is very unfavorable. We report an operated case of the disease with rapid growth of pulmonary metastasis. The silver, vimentin and factor VIII stainings were available for diagnosis of this disease. The 46-year-old female patient died of bleeding from the pulmonary metastasis eight months after the operation without other distant metastasis. Even if a lung tumor is pointed out and thought to be benign, frequent follow up is necessary. Furthermore, we recommend positive lung biopsy by thoracotomy or video-assisted-thoracic-surgery.

[Two cases of metastatic lung tumor from leiomyosarcoma of the uterus].

Three pulmonary resections were performed for two cases of metastatic lung tumor from uterine leiomyosarcoma. Case 1 was diagnosed as myoma uteri. Preoperative chest X-ray examination showed an abnormal shadow in the left upper lobe. Postoperative diagnosis was leiomyosarcoma. During radiotherapy for the pelvic lesion, the lung tumor was growing very rapidly and micrometastases were detected bilaterally without distant metastasis. Bilateral pulmonary resections were performed. Three months and 15 days later, second left pulmonary and thoracic wall resections were performed because of lung and pleural recurrence. Seven months after the first pulmonary operation, she died of systemic metastases. Case 2 was admitted to the hospital because of genital bleeding and diagnosed leiomyosarcoma of the uterus. The patient had a metastatic lung tumor in the left lower lobe preoperatively. Eighteen days after hysterectomy, left lower lobectomy was performed because progressive hemoptysis occurred for rapidly enlarged metastatic lesion. She died of intrapelvic bleeding three months after the operation without respiratory symptom. The prognosis of synchronously found out lung metastasis from uterine leiomyosarcoma is very poor. Neither chemotherapy nor radiotherapy is effective. Although it's difficult and controversial to decide the surgical indication, we need to operate on as soon as possible when there is no other distant metastasis. Even if the timing of operation is late, we can find the surgical significance of pulmonary resection for reduction of respiratory symptoms and probability of longer survival.

[A case of bronchogenic cyst in the posterior mediastinum with high level of serum SCC antigen].

A 78-year-old man was picked up by chest X-ray findings. His CT scan film showed a posterior mediasinal tumor shadow. Serum level of SCC antigen was high and thoracotomy revealed bronchogenic cyst with high production of SCC antigen. After resection of the cyst, his serum level of SCC was within normal range. It is proposed that the elevated serum level of SCC is due to chronic inflammation or structural feature of the cystic wall.

[Low-dose methotrexate and sequential 5-FU treatment in advanced gastric cancer].

Combination chemotherapy of low-dose methotrexate and sequential 5-FU was given to 16 patients with advanced gastric cancer. Treatment regimen : methotrexate 30 mg/m2 followed 3 hours later with 5-FU, 750 mg and 24 hours later with leucovorin 30 mg/m2. Each drug was administered as a bolus. Treatment was repeated weekly. The median ECOG PS was 3 (range 2-4) and median age was 55.1 years (range 28-87). Of these 16 patients who were evaluated, four had PR, and 5 had MR, giving a response rate of 25%. Of the 7 patients who had been previously treated with a combination of chemotherapy including 5-FU, the figures were 2 PR and 3 MR. This regimen has been well tolerated by the patients. No patient had a WBC nadir of less than 2000 cells/mm3, and no patient had a platelet count nadir of less than one hundred thousand platelets/mm3. No other toxicity was detected.

[Efficacy of combination therapy consisting of cyclophosphamide, adriamycin, UFT and oophorectomy/tamoxifen for advanced or recurrent breast cancer].

Combination therapy consisting of cyclophosphamide (CPA), adriamycin (ADR), UFT and endocrine therapy of oophorectomy or tamoxifen (TAM) was given female patients with advanced or recurrent breast cancer. Premenopausal patients initially underwent oophorectomy, then were administered 300 mg/day of CPA and 30 mg/day of ADR intravenous injection every 2 weeks, and 300 mg/day of UFT orally every day. Postmenopausal patients, on the other hand, were administered the same three chemotherapeutic drugs and 30 mg/day of TAM orally every day instead of receiving castration. A total of 10 patients were registered, and nine of them were eligible. Premenopausal cases were six, and postmenopausal cases were three. The objective response rates were 50% (3/6) for premenopausal women and 33% (1/3) for postmenopausal women. Complete response was observed in one patient and partial response in three patients. The therapy was effective for lymph nodes (2/2), lung (3/4), breast (2/3), skin or subcutis (1/2), but it showed no effect on bone, pleural effusion or ascites. The overall 5-year survival rate was 53.3%. The main side effects of more than Grade 1 were leucopenia 33% (3/9), anorexia 22% (2/9) and malaise 22% (2/9). They were relatively mild for therapy involving ADR. This therapy was considered useful for advanced or recurrent breast cancer.

[A case of locally advanced breast cancer successfully treated with wide resection and reconstruction of chest wall with A-O metallic plates].

A 63-year-old female with locally advanced breast cancer was treated with preoperative chemotherapy using docetaxel. The therapeutic regimen was comprised of four cycles at 3-week intervals. One cycle consisted of 80 mg of docetaxel which was administered on day 1. A remarkable response was confirmed. The side effects such as leukopenia, general fatigue and alopecia were moderate and had no influence on the patient's QOL. After preoperative chemotherapy, a full thickness chest wall resection was performed. Chest wall defect was reconstructed with orthopedic A-O metallic plates, Marlex mesh and rectus abdominis myocutaneous flap. These metal plates were very useful because it was easy to bend and twist them manually to fit the defect at the time of operation. Moreover, the curved metal plates preserved the cone form of the chest cage. The postoperative course was favourable without frail chest or wound infection.

[Partial liver resection with small skin incision for multidisciplinary treatment of hepatocellular carcinoma].

Hepatocellular carcinoma (HCC) is likely to be detected as multiple intrahepatic metastases, and trans-arterial embolization (TAE) is the treatment of choice. However, other therapy is needed when TAE is not effective for a tumor. We performed partial liver resection with small skin incision in two such patients. One patient had two HCCs of segment 6 (S6) and S7, and TAE was performed twice. However, HCC of S7 was viable, and we partially resected the tumor of S7. In the other patient, HCC was multiple in segment 6 and 7. TAE was performed three times, and in segment 6 we detected a tumor which grew into the extrahepatic space and seemed uneffected by TAE. We performed a partial resection of liver segment 6. After 3 months, TAE was repeated in the latter. The operation time was about three hours, and no blood transfusion was performed. Two of the patients were discharged on the 13th postoperative day. We concluded that partial liver resection with small skin incision was useful for one case of multidisciplinary treatment for hepatocellular carcinoma.

[A long-survival case of unresectable liver metastasis of colon cancer by hepatic arterial intermittent multidrug infusion chemotherapy].

We experienced a long-survival case with unresectable liver metastasis of sigmoid cancer by hepatic arterial intermittent multidrug infusion chemotherapy. The patient was a 65-year-old man. A movable tumor was palpable at the left lower abdomen. The serum CEA value was 15.6 ng/ml. Abdominal CT scan showed a 3 cm-sized low-density area near the root of the right and middle hepatic vein in segment 4. He had undergone sigmoidectomy, cholecystectomy and cannulation into the proper hepatic artery. Chemotherapy was performed by using 5-FU (250 mg), Adriacin (5 mg) and mitomycin C (4 mg) every second week from 4 months after surgery as an outpatient. In the CT scan image, the low-density area completely disappeared at 10 months after treatment, and the CR interval was 14 months. The patient has been alive for 5 years after surgery.

[Phase II study of 5'-DFUR (5'-deoxy-5-fluorouridine) by the Cooperative Study Group].

Phase II study of 5'-DFUR was conducted in 195 patients with malignant tumors by the Osaka Chemotherapy Cooperative Study Group. Five CR and 20 PR cases were obtained out 133 evaluable cases and the total response rate was 18.8% (15.8% in gastric, 38.1% in breast) One PR was observed for each of head & neck and esophageal cancer. It was noteworthy that four CR were observed in breast cancer. Adverse reaction was observed in 61 out of 151 cases (40.4%), and major side effects were digestive symptoms such as diarrhea (22.5%), nausea-vomiting (11.9%) and anorexia (10.6%). These results suggest that 5'-DFUR can be useful for the treatment of malignant tumors.

[Microwave tissue coagulation therapy with mini-laparotomy for hepatocellular carcinoma].

Recently microwave tissue coagulation therapy (MCT) has been established as a therapeutic procedure for hepatocellular carcinoma (HCC). The route of approach to penetrate the tumor is two-fold: By the percutaneous route (PMCT) or by conventional laparotomy. PMCT has the advantage of less invasive therapy, however, it is not applicable to a tumor that cannot be detected by ultrasonography. Therefore, we tried MCT with mini-laparotomy. The patient was a 57-year old man with HCC located in the S6 subsegment. The operation was performed at the semi-right side up position, and laparotomy was done by skin incision of 12 cm in the right hypochondral area. MCT was performed about 30 times, with an output of 60 watts for 45 seconds at a time. Abdominal CT image showed a low-density area of 7 x 4 cm on the 18th postoperative day. Slight damage to the liver and no obvious complications were observed. We concluded that a much wider range-coagulation than PMCT was able to be obtained by MCT with mini-laparotomy, and postoperative quality of life was better than MCT with conventional laparotomy.

[Hepatic arterial chemotherapy for liver cancer over a period of 8 years].

Hepatic arterial chemotherapy was performed for 27 patients with primary (3), metastatic liver cancer (21), and 3 other cases, over a period of 8 years. Chemotherapy was performed by intermittent hepatic arterial infusion of 5-FU or FAM (in case of metastatic tumor from colorectal cancer), FAM (from gastric cancer), and CDDP or Farmorubicin (HCC). Hepatic resection was performed in 10 cases of metastatic tumor from colorectal cancer, and 8 cases of 10 were curative operation. The 5-year survival rates of curative liver resection group, and non-curative liver resection or non-resection group were 57.1% and 12.5%, respectively. As is the case with metastatic cancer from gastric cancer, pancreatic cancer, and hepatocellular carcinoma (HCC), the prognosis was poor except for one CR case of HCC. We concluded that hepatic arterial chemotherapy may be recommended for a curative resected case of liver metastasis from colorectal cancer.

[A case of liver metastasis of gastric leiomyosarcoma successfully treated by transarterial hepatic chemo-embolization and intra hepato-arterial chemotherapy repeated with infusion-a-port].

The patient was a 68-year-old male, who underwent total gastrectomy for giant leiomyosarcoma of the stomach and then had multiple hepatic metastases one year and six months later. Thus, transarterial hepatic chemo-embolization therapy with Lipiodol, adriamycin and gelfoam was given. Moreover, using a reservoir catheter and infusion arterial port, intermittent arterial infusion therapy with adriamycin, cyclophosphamide, and vincristine was attempted. In the metastasis lesion where there were rich blood vessels, Lipiodol was accumulated and the tumor was reduced on abdominal CT. The result indicated the efficacy of this treatment.

[The significance of neoadjuvant intra-arterial chemotherapy for locally advanced breast cancer by different methods].

For neoadjuvant intra-arterial (IA) chemotherapy in locally advanced breast cancer patients, Seldinger's methods were found to be convenient and had the same effect and outcome as conventional methods. The prognosis of the patients in whom IA chemotherapy was locally effective and had fewer than n 1 lymph node metastases was comparatively favorable. However, several patients who underwent IA chemotherapy later experienced local recurrence, and the cause of these patient's death was distant metastases in almost all cases. We recommend neoadjuvant IA and systemic chemotherapy, and systemic adjuvant chemotherapy.

[Neoadjuvant intraarterial chemotherapy for ten cases of inflammatory breast cancer by Seldinger's methods].

The conventional methods (CM) of intraarterial (IA) chemotherapy for inflammatory breast cancer were catheterizations from superior epigastric and brachial artery under general anesthesia. Since 1997, we selected the Seldinger's methods (SM) for ten cases of the disease to control local effects and simplify the technique. Complications of the SM were slight, and side effects were equal to those with CM. The postoperative pathological findings of the SM showed the direct effects of chemotherapy to tumor cells (degenerative and necrotic changes) as compared with the embolism-like effects of CM. But the overall histological effects of chemotherapy by SM were almost equal to those for CM. The strong points of the SM were as follows: 1) More selective IA chemotherapy is available, 2) one can find the passage of the aim vessels and no trouble related to catheter, 3) general anesthesia is not necessary, and the techniques are simple, 4) the wounds are not remarkable. The disadvantages are as follows: 1) Patients must rest the day of IA chemotherapy, 2) in 20% of the procedures, one can not search the vessels, and 3) in 4 cases complications of stiffness of Mj or Mn pectral muscles were found. In future, we expect more effective results by dose escalation or combination chemotherapy.

[A case report of successful transcatheter arterial embolization therapy for osseous metastasis of hepatocellular carcinoma].

A 74-year-old man was admitted to our hospital with a chief complaint of severe local pain of the hip joint. Radiological findings showed a metastasized lesion on the left side of the pelvic wall originated from hepatocellular carcinoma (HCC) in the anterior segment of the liver. Transcatheter arterial embolization (TAE) therapy using epirubicin, Lipiodol and Spongel was successfully performed twice for primary HCC, and four times for osseous metastasis of HCC. After TAE therapy, the size of the metastasized lesion decreased with relief of pain, and an improvement in performance status of 4 to 2 was achieved. In conclusion, TAE therapy is thought to be very useful in the treatment of osseous metastasis of HCC with severe local pain.