Minimally invasive transabdominal collection of preimplantation embryos from the common marmoset monkey (Callithrix jacchus).

A novel, minimally invasive, transabdominal embryo collection method (transabdominal method) was developed as an alternative to a standard abdominal incision for embryo collection in the common marmoset. The abdominal incision method was used for 304 flushes using 36 female animals, whereas the transabdominal method was used for 488 flushes using 48 females; successful embryo collection rates were 48.0% and 48.4% (P > 0.05), respectively. These techniques were successfully duplicated at another institute (German Primate Center, DPZ). At that institution, successful embryo collection rates were 88.9% and 77.8% for the abdominal incision and transabdominal methods, respectively (P > 0.05), whereas the average numbers of preimplantation embryos obtained per flush were (mean ± SD) 1.91 ± 0.35 and 1.71 ± 0.14 (P > 0.05). The transabdominal method reduced animal stress, did not require incisional wound healing, and enabled successive embryo recoveries to be done much sooner. More embryos in early developmental stages (zygotes/morulae) were recovered using the transabdominal method (76.1%) than the abdominal incision method (52.6%, P < 0.01). In contrast, recovery of arrested or abnormal embryos was not significantly different between these two methods (9.8% and 8.3%). To verify developmental ability of embryos recovered by the transabdominal method, transfer of 28 normal embryos to 14 surrogate mothers yielded a nidation rate of 57%. Five females sustained term pregnancies and eight neonates were born. This novel transabdominal method will facilitate progress in marmoset developmental biology and embryology.

Progress reports on immune gene therapy for stage IV renal cell cancer using lethally irradiated granulocyte-macrophage colony-stimulating factor-transduced autologous renal cancer cells.

There is no effective treatment for patients with stage IV renal cell cancer (RCC), although the introduction of new therapy is imminent. Cancer gene therapy is currently considered to be one of the most promising therapeutic modalities in the field of cancer treatment. Based on the results of animal studies, vaccination using autologous granulocyte-macrophage colony-stimulating factor-transduced renal cancer cells appears promising. Before initiating a clinical study using an ex vivo gene-transduced autologous cell vaccine-based immunogene therapy for RCC in Japan, in 1992 we initially planned a Japanese version of a clinical protocol in collaboration with a US group. In 1993, the original protocol was refined. We performed five preclinical qualification studies using RCC nephrectomy specimens from patients in 1997, and the results showed that preparation of RCC cells for autologous vaccines at the Clinical Cell Technology Facility, Research Hospital of the Institute of Medical Science, University of Tokyo, was feasible. Subsequently in August 1998, the Ministry of Health and Welfare and the Ministry of Education, Science, Culture, and Sport approved our clinical protocol. We have recruited two patients with stage IV RCC to our study so far. Here we report the background to the initiation of cancer gene therapy in Japan.

Primary carcinoid tumor in a horseshoe kidney.

A case of primary carcinoid tumor arising within a horseshoe kidney in a 51-year-old woman is reported. The tumor was found incidentally by computed tomography (CT) during a check-up for a suspected gall bladder polyp. Histologic, immunohistochemical and electron microscopic analyses of this tumor revealed features typical of carcinoid tumor. Primary carcinoid tumor of the kidney is extremely rare and only 32 cases were previously reported, including five cases in horseshoe kidneys. None of these five cases in horseshoe kidneys demonstrated any evidence of local or distant metastases and all were alive at the time of reporting without evidence of disease after up to 3 years of follow up. The present case, even with accompanying lymph nodal metastasis, also has had no evidence of local recurrence or distant metastasis for 3 years post operation. Primary carcinoid tumor arising within horseshoe kidneys appear to be more benign than those within non-horseshoe kidneys.

Interferon therapy for renal cell carcinoma in hemodialysis patients: report of two patients.

The method of interferon therapy has not been clarified in hemodialysis patients with renal cell carcinoma. Two hemodialysis patients with renal cell carcinoma were treated with natural interferon-alpha. The serum levels of interferon-alpha in both patients were measured after 24, 48, 72 and 96 h of injection. In case 1, the serum concentration of interferon-alpha after 24 h of injection at a dose of 3 x 10(6) reached a maximum and decreased gradually. In case 2, the serum concentration of interferon-alpha reached a maximum 24 h after injection at a dose of 3 x 10(6) and decreasing gradually after this. After an injection of interferon-alpha at a concentration of 6 x 10(6) 7 days later, the serum concentration of interferon-alpha reached a maximum 48 h after the injection and decreased gradually thereafter. The serum interferon-alpha concentrations of these cases were higher than normal renal function patients in other literature. It is necessary to modify the interval between injections for hemodialysis patients with renal cell carcinoma.

Total replacement of the aorta and the inferior vena cava after resection of an extra-adrenal malignant pheochromocytoma.

Extra-adrenal malignant pheochromocytomas are difficult to operate on because of a tight adhesion to the circumference of tumors. However, an operation is the most curable treatment. Sometimes a replacement of the inferior vena cava with grafts is necessary for removing tumors completely. We totally replaced the aorta and the inferior vena cava after resection of an extra-adrenal malignant pheochromocytoma. No recurrence has been detected, and the aortic graft has had a good patency after 36 months postoperatively. But the vena caval graft has had no patency after 21 months postoperatively.

Collection of peripheral blood stem cells with granulocyte-colony-stimulating factor alone in testicular cancer patients.

BACKGROUND: High-dose chemotherapy with the transplantation of peripheral blood stem cells (PBSC) has been performed for the treatment of advanced testicular cancer patients. Recently, it has been reported that, in healthy donors, a large quantity of stem cells can be transferred to peripheral blood using granulocyte-colony-stimulating factor (G-CSF) alone. Therefore, it was decided to try to harvest PBSC from three patients having testicular cancers with G-CSF alone. METHODS: The three patients with testicular cancer were 26, 56 and 62-years-old. They had undergone five, two and three cycles of chemotherapy, respectively, but no radiation therapy. Granulocyte colony-stimulating factor was subcutaneously injected (250 microg) into each patient twice per day for 6 days. Peripheral blood stem cells were harvested for 3 days (days 4-6) and mononuclear cells (MNC), CD34-positive cells and colony-forming units of granulocyte-macrophage (CFU-GM) in PBSC collected by apheresis were measured. RESULTS: Apheresis showed that the total MNC count was 20.2 x 10(8)/kg (range, 10.6-25.9 x 10(8)/kg), the CD34-positive cell count was 0.98 x 10(6)/kg (range, 0.75-1.4 x 10(6)/kg) and the total CFU-GM count was 1.36 x 10(5)/kg (range, 0.25-3.0 x 10(5)/kg). CONCLUSION: After mobilization of peripheral blood stem cells with G-CSF alone, sufficient amounts of MNC were obtained from testicular cancer patients who had undergone chemotherapy several times. However, sufficient amounts of CD34-positive cells and CFU-GM could not be obtained. These results suggested that the G-CSF dose was not adequate for harvesting sufficient amounts of CD34-positive cells and CFU-GM.

Development of black gallstones after the nonsurgical management of splenic injury: report of a case.

A 22-year-old man was admitted to our Emergency Department after suffering splenic injury in a traffic accident. His intraabdominal bleeding was treated nonsurgically by the administration of total parenteral nutrition (TPN) and blood transfusions of packed red cells. He presented again 2 months after his discharge, being 3 months after the injury, for right hypochondralgia, at which time a gallstone was demonstrated on ultrasound (US) and computed tomography (CT). After endoscopic laparoscopic cholecystectomy, his symptoms disappeared and he has remained well since. The clinical course of this patient indicates that hemolytic hyperbilirubinemia can cause black gallstones as a late complication of the nonsurgical management of abdominal blunt trauma.

Detection of interleukin-1 activity in human bladder cancer cell lines.

Detection of interleukin (IL)-1 activity was studied in two human bladder cancer cell lines, T24 and EJ1, and one rat bladder carcinoma cell line, 804G. Significantly high proliferation of mouse thymocytes in the assay of IL-1 activity was observed in the conditioned medium (CM) of T24 cells, indicating that the cells released IL-1-like activity. Enzyme-linked immunosorbent assay (ELISA) and Northern blot analysis showed the presence of both IL-1 alpha and IL-1 beta in the CM of T24 cells and expression of mRNAs of both cytokines in the cells. Interleukin-1 activity in EJ1 cells, which produced a little activity, was induced by E. coli lipopolysaccharide (LPS) while it was not induced in T24 by either LPS or other test substances. Conditioned medium of T24 increased proliferation of both T24 and EJ1 in cell-growth assay. Further investigation of the mode of action and role of cytokines, especially those from tumor cells themselves, is necessary in relation to BCG or photodynamic therapy.

[Intralesional and transarterial administration of interferon for renal cell carcinoma].

Topical administration of interferon for locally recurrent renal cell carcinoma in 2 cases was reported. Case 1: a 56-year-old woman had undergone radical nephrectomy for left renal cell carcinoma. She had a locally recurrent tumor adjacent to spleen 26 months after the surgery. She received 3 million units of interferon alpha every day intralesionally for almost 3 years without any distant metastasis. Case 2: a 59-year-old man had undergone left radical nephrectomy for renal cell carcinoma. He had recurrent tumor contiguous to spleen 24 months after nephrectomy. With Seldinger method a catheter was indwelled selectively in splenic artery. Two million units of interferon gamma was administered through the catheter, twice a week for 4 weeks, and once a week in subsequent course. The tumor showed necrosis on CT film. He died of DIC 5 months after the initiation of intraarterial administration of interferon gamma. Topical use of interferon showed some favorable effects in both cases even after the failure of systemic administration. Further investigation is warranted to demonstrate the advantage of topical use of interferon over systemic administration.

[Significance of urinary cadmium concentration as an indicator of cadmium exposure in a population exposed to cadmium in a general environment].

Study was made on the significance of urinary cadmium concentration as an indicator of internal dose in a general environment polluted by cadmium. As an index of external dose, the mean rice cadmium concentration in each of 23 villages was employed. Morning urine samples were collected from 3,178 inhabitants of 23 cadmium-polluted villages of Kakehashi River basin in Ishikawa Prefecture and 294 inhabitants of two nonpolluted villages. Cadmium concentration in urine was determined. In addition, their residential history and intake of cadmium-polluted rice were obtained. All the subjects were 50 years of age or over. Participation rates were 91% for the polluted area and 94% for the nonpolluted area. Urinary cadmium concentrations were higher in the inhabitants of the cadmium-polluted area than in those of the nonpolluted area. Among the inhabitants in the cadmium-polluted area, urinary cadmium concentrations were higher in the subjects who had consumed cadmium polluted rice than in those who had not. The mean urinary cadmium concentrations in each village increased proportionally to increase in mean village rice cadmium concentration (intensity of exposure) when the inhabitants were classified according to period of residence at their present address. The mean urinary cadmium concentrations in each village were also significantly correlated with period of residence at the present address (duration of exposure) when the inhabitants were classified according to mean rice cadmium concentration of their village. It is concluded that urinary cadmium concentration mainly reflects the body burden of cadmium in a general environment polluted by cadmium.

[Adrenal myelolipoma: a report of a case].

This is a case report of an adrenal myelolipoma, accidentally diagnosed during a work-up for bladder tumor. A 67-year-old male presented with the chief complaint of gross hematuria. He was subsequently diagnosed as having a bladder tumor, which was resected transurethrally and was found to be a transitional cell carcinoma of Grade 2 and Stage pT2. During further examination for metastasis computed tomography (CT) scan revealed a round tumor (approximately 5 cm in diameter) in the left adrenal. A tentative diagnosis was reached based on the scan, and surgery was undertaken to remove the tumor. A well-encapsulated tumor, yellowish and partly dark brown in color and 60 grams in weight, was retrieved. The tumor consisted of mature lipoid cells with myeloid cells scattered among them which verified the pathological diagnosis of adrenal myelolipoma. The present case is the 16th clinical case of adrenal myelolipoma reported in the Japanese literature.

[Adjustment of urinary delta-aminolevulinic acid concentrations in workers exposed to lead and heat. Adjustments of specific gravity to 1.020 and urinary volume coefficients].

Adjustments of urinary ALA concentrations as to urinary specific gravity and creatinine were examined for workers exposed to lead and heat. Judging from our findings, we suggested that it was preferable to adopt the specific gravity (UG) at 1.020 as the adjustment value to obtain the correct urinary ALA concentration. Though corrected values thus obtained were found adequate for urine in the normal range of specific gravity, they failed to be adequate for concentrated urine samples higher than UG 1.025. Urinary volume adjustment was found to be necessary for these concentrated urine in stead of urinary specific gravity adjustment. For the practical purposes, we postulated urinary volume coefficients, which were estimated to be 0.5 for samples ranging from UG 1.026 to 1.030, 0.4 for samples from UG 1.031 to 1.035 and 0.3 for samples from UG 1.036 to 1.040, respectively.