Analyses of T-cell differentiation from hemopoietic stem cells in the G0 phase by an in vitro method.

Using differential radiation sensitivity of components of mouse embryonal thymus, an in vitro method for studying T-cell differentiation from hemopoietic stem cells (HSCs) in the G0 phase was established. Intrathymic T-cell precursors present in embryonal thymus were found to be quite radioresistant (up to 20 Gy), and consequently 25-Gy-irradiated embryonal thymic lobes were used. Thymic lobes (25-Gy irradiated) taken from mouse fetuses (gestation day 15) were placed in Millipore-HA culture plates supported on squares of gelatin foam sponge in 24-well culture plates in which neonatal thymus stromal cells were cultured. HSCs (10(5) cells per well) in the G0 phase were added to these thymic lobes and cocultured at 37 degrees C in a 5% CO2/95% air incubator. Half the culture medium was changed every week. After 3 weeks, a large number of colonies had formed. Immunohistochemical studies and fluorescence-activated cell sorter analyses revealed that the colonizing cells regularly develop and exhibit surface markers characteristic of T cells (Thy-1, IL-2R, L3T4, Lyt-2, etc.). In situ hybridization analyses revealed that mRNA expression for T-cell receptor (TCR) beta chains occurred within colonizing cells. Using a monoclonal antibody (F23.1), expression of TCR beta-chain variable domain (V beta 8) on the surface of these developing T cells was demonstrated. These cells responded to interleukin 2 and/or anti-CD3 monoclonal antibody, indicating functional T cells. This method will be useful in studying T-cell differentiation pathways from pluripotent HSCs and in clarifying the mechanisms involved in negative and positive selection of T cells within the thymus.     

Anomeric preference in uptake of D-glucose and of D-galactose by rat lenses

To determine the anomeric preference of uptake of D-glucose and of D-galactose by rat lenses, we crystallized alpha-, beta-D-[U-14C]glucose (720 microCi/mmol) and alpha-, beta-D-[U-14C]galactose (180 microCi/mmol) by our method and incubated them separately with rat lenses for 1 min, because of the short half-life of mutarotation of alpha-D-glucose (9.6 min) and of alpha-D-galactose (4.6 min) in HEPES medium at 30 degrees C. During aerobic incubation of rat lenses in HEPES medium containing radioactive alpha or beta anomer of D-glucose, there was no significant difference in the rate of uptake between alpha and beta anomers of D-glucose by rat lenses. However, 1.59 times greater incorporation of alpha-D-galactose was observed over that of beta-D-galactose under the same conditions.     

A case of immunohistochemically carcinoembryonic antigen producing cervical cancer--the evaluation of plasma CEA levels in 201 patients with uterine cancer

A case of a cervical cancer in a 40-year-old woman with a plasma carcinoembryonic antigen (CEA) level of 29.4 ng/ml was investigated using light and electron microscopy. In addition, the plasma CEA levels before treatment were determined in 168 patients with cervical cancer and in 33 patients with endometrial cancer. CEA was found to be elevated in the plasma of 19% of those with cervical cancer and in 6% of those with endometrial cancer. Effective serial plasma CEA determinations following therapy, in patients whose plasma or tumors initially contain elevated amounts of antigen, might be useful as an adjunctive method in the earlier detection of a recurrent cancer.     

Clinical similarities of hereditary progressive/dopa responsive dystonia caused by different types of mutations in the GTP cyclohydrolase I gene

OBJECTIVE—Hereditary progressive dystonia withpronounced diurnal fluctuation ((HPD)/dopa responsive dystonia (DRD))is a childhood onset dystonia which responds to levodopa. Variousclinical signs and symptoms of HPD/DRD have been recognised to date.Mutations in the GTP cyclohydrolase I (GTP-CH-I) gene were recentlyidentified as the cause of HPD/DRD. In the present study, the GTP-CH-Igene and the clinical features of eight HPD/DRD patients from sixfamilies were analysed to determine the correlationsbetween clinical expression and the mutations in the GTP-CH-I gene.
METHODS—The exons, exon-intron junctions, and anindispensable part of the 5' flanking region of the GTP-CH-I gene weresequenced in the eight clinically diagnosed patients with HPD/DRD andtheir asymptomatic parents.
RESULTS—Three independent mutations in theGTP-CH-I gene were found in three patients. One of the patients and herasymptomatic mother were heterozygous for a novel mutation at theinitiation codon. The three patients with dissimilar GTP-CH-I mutationsexhibited similar clinical features. The other five patients withnormal sequences presented several features not manifested by the three patients with the mutations. No mutation was found in the 5' flanking region of any patients or their parents.
CONCLUSIONS—A novel initiation codon mutation wasfound in a Japanese patient with HPD/DRD. The clinical manifestationscommon to the patients with HPD/DRD with a mutated GTP-CH-I gene werealso identified. Although focal manifestations of HPD/DRD associatedwith the mutations of this gene will be broadened, it is inferred thatthese clinical features are fundamental to HPD/DRD caused by mutationsin this gene.

     

Biliary cystadenoma with mesenchymal stroma of the liver: Correlation between unusual MR appearance and pathologic findings

We reported a case of the biliary cystadenoma of the liver. The cystic mass had labulation and septation and showed marked hyperintensity on T1-weighted images and hypointensity on T2-weighted images; MR findings were very unusual for cystadenoma. The content of the cystic mass was jelly-like, thick mucinous fluid without intracystic hemorrhage. We concluded that these unusual signal intensities of the cyst were due to hyperproteinous mucinous fluid.     

Development of new immunoradiometric assay for CA 125 antigen using two monoclonal antibodies produced by immunizing lung cancer cells

CA 125 is an antigen associated with non-mucinous epithelial ovarian cancer, which is defined by OC 125 antibody developed by immunizing ovarian cancer cells. We have produced two monoclonal antibodies, 130-22 and 145-9, by using the human lung adenocarcinoma cell line PC-9. Both 130-22 and 145-9 antibodies recognized CA 125 antigen. However, the binding sites seemed to be separate from those of OC 125. Testing by 9 immunoradiometric assays (IRMA), using different combinations of the 3 monoclonal antibodies 130-22, 145-9 and OC 125 demonstrated that the best standard curve for detecting CA 125 could be obtained by a "simultaneous sandwich" assay based on a mixture of 125I-labeled OC 125 and 130-22 or 145-9 coated beads. One-step IRMA, using 130-22 as a tracer and 145-9 as an immunoadsorbent, also showed good reproducibility and sensitivity for measuring CA 125. Antigens were detectable in the culture supernatants of PC-9 cells and 5 of 6 ovarian cancer and endometrial adenocarcinoma cells. These results indicate that one-step IRMA using 130-22 and 145-9 is useful for detecting CA 125 antigen.     

A case of a probable "cancer family syndrome" presenting four synchronous malignancies of the colon

A 72-year-old male was admitted because of right lower quadrant pain, Barium enema and total colonoscopy disclosed multiple colon cancers and sequentially, a subtotal colectomy was performed. The resected specimen demonstrated 3 advanced carcinomas and an adenomatous cancer with additional multiple polyps. Investigation of his family history revealed that his mother and his elder sister had died of uterine cancer, and that his elder brother, his nephew, and his niece had been operated on for colorectal cancer. We thus supposed a case of "Cancer Family Syndrome" presenting multiple neoplasms of the colon.     

Second generation shock wave lithotripsy: experience with the Siemens Lithostar

In 1980, extracorporeal shock wave lithotripsy (ESWL) was incorporated as a nonsurgical method of stone removal in the cases of nephrolithiasis and rapidly found worldwide acceptance. Several devices commonly designated "second generation" lithotripters vs "first generation" Dornier HM3 are now under experimental or clinical trial. We report our clinical experience of ESWL using a Siemens Lithostar and compared it with that obtained using a Dornier HM3. One hundred patients were treated during the period of April through October, 1986 using an HM3, and 100 other patients were treated using a Lithostar from April to August, 1988. More cases were treated with a Lithostar than with a HM3. Nearly 10% of all patients treated by ESWL required additional therapeutic approaches (excepted ureteral stent) either with HM3 or Lithostar. However, in the cases of ureteral stone, with the Lithostar more cases required adjuvant procedures (TUL) than HM3. Significantly more shock waves were needed with Lithostar than HM3 for complete fragmentation of the same size of renal and ureteral stones. The stone-free rate during a one month period after ESWL was nearly the same for HM3 and Lithostar (HM3: 84.3%, Lithostar: 83.5%). Lithostar is a multifunctional lithotriptor which has most of the advantages required by the lithotripter.