Biomedical subjects

J Hata

Publications and source records attributed to J Hata.

At least 19 recordsLinked to original sources

PubMed1997

A neuroblastoma cell line derived from a case detected through a mass screening system in Japan: a case report including the biologic and phenotypic characteristics of the cell line.

BACKGROUND: A mass screening system in Japan, which involves measuring urinary catecholamine metabolites, has resulted in an increasing number of cases of neuroblastoma, most of which have favorable biologic properties and some of which are associated with tumor regression or involution. At the time this study was begun, the characteristics and biologic nature of the neuroblastomas had not been fully defined, because a cell line had not yet been established with tumor tissue taken from a neuroblastoma detected in the mass screening. METHODS: The authors established a cell line by tissue culture for over 50 generations from a neuroblastoma found during mass screening, which was characterized by favorable histology, with a triploid DNA stemline and without N-myc gene amplification. The morphologic and biologic characteristics of the new cell line were investigated in vitro. RESULTS: The cell line, designated MASS-NB-SCH-1, has neuronal properties, such as neurite-like processes and neurofilaments, as well as the expression of vimentin and fibronectin in studies of the cell morphology and immunohistochemistry. Karyotype analysis detected the presence of 42-46 chromosomes, with a deletion of the short arm of 1 of the 3 copies of chromosome 1. DNA ploidy was near-diploid in association with 20-fold amplification of N-myc genes. CONCLUSIONS: The cell line has a nature distinct from the original tumor tissue. It may be characterized by phenotypic change caused by clonal selection or evolution of aggressive, proliferative properties in vitro. This cell line will be a useful model to investigate the properties of the neuroblastoma in relation to the N-myc amplification mechanism.

Biology↗

PubMed1997

Neurogenic potential of Ewing's sarcoma cells.

We investigated the capacity of eight well-characterized Ewing's sarcoma cell lines to differentiate towards a neural phenotype. Ewing's sarcoma cells expressed the neuroectoderm-associated antigens such as nerve growth factor (NGF) receptor, N-CAM (6H7 and Leu-19) and Leu-7. Ewing's sarcoma cells also exhibited the potential for neural differentiation at the mRNA level; neuron-specific medium- and low-sized filament (NF-M and NF-L) expression was induced by dibutyryladenosine cyclic monophosphate. The pattern of expression of NF-L obtained by using alternative polyadenylation sites in Ewing's sarcoma cells differed from that in peripheral primitive neuroectodermal tumour (PNET) cells, and was similar to that in undifferentiated neural tissues. Furthermore, the NGF receptors detected by immunohistochemistry were found to be non-functional as assayed by c-fos induction with NGF treatment. The results indicate that Ewing's sarcoma cells maintain a primitive phenotype and have the potential to differentiate into a neural phenotype, indicating that Ewing's sarcoma is distinct from PNET.

Bone Neoplasms↗

PubMed1997

Effects of octreotide, a somatostatin analogue, on gastric function evaluated by real-time ultrasonography.

BACKGROUND: Somatostatin exerts inhibitory effects on physiological functions in the gastrointestinal tract. The actions differ, however, depending on the test meal, dose, and other factors. AIMS: To determine by use of ultrasonography and scintigraphy the effect of a somatostatin analog, octreotide, on gastric emptying and antral contraction. SUBJECTS: Twenty healthy men; mean age 23.9 years METHODS: Subjects were studied for 7 days, once after subcutaneous injection of octreotide, 50 microg, 5 min before the ingestion of a test meal and once after subcutaneous injection of placebo. Ten subjects received a liquid meal, 10 others received a solid meal. With the liquid meal, gastric emptying was measured 15 min after its ingestion and antral contraction was measured for 15 min by ultrasonography. With the solid meal, gastric emptying was measured both by ultrasonography (n = 10) and by simultaneous scintigraphy (n = 6), with antral contraction measured by ultrasonography for 5 h after ingestion of the meal. RESULTS AND CONCLUSIONS: Octreotide given with a liquid or solid meal inhibited gastric emptying in healthy subjects. A significant suppression of antral contraction occurred only with a solid meal.

Adult↗

PubMed1997

Impaired male sex development in an infant with molecularly defined partial 9p monosomy: implication for a testis forming gene(s) on 9p.

This paper describes a genetically male infant with impaired male sex development and partial 9p monosomy. The external genitalia were ambiguous with microphallus (penile length at birth 10 mm, mean age matched normal length 29 mm (SD 5)), hypospadias, and hypoplastic scrotum. The tests were undescended and severely hypoplastic (testis size at 12 months of age, right 8 x 5 x 4 mm and left 4 x 3 x 2 mm; mean age matched normal size, length 18 mm (SD 2), width 11 mm (SD 1). Cytogenetic studies showed a 46,XY,del(9)(p23) karyotype in all the 30 peripheral lymphocytes and 20 skin fibroblasts examined. Microsatellite analysis for a total of 13 loci assigned to the 9p22-24 region showed that the deleted chromosome 9 was of paternal origin and was missing a region distal to D9S168. Southern blot analysis for D9S47 also confirmed the 9p deletion. The sequence of SRY was normal. The results provide further support for the previously proposed hypothesis that a gene(s) for testis formation is present on the distal part of 9p and indicate in molecular terms that the putative testis forming gene(s) resides in the region distal to D9S168.

Blotting, Southern↗

PubMed1997

Fatal diffuse alveolar damage from bird fanciers' lung.

A 68-year-old man, who had continuing exposure to budgerigars, developed fatal acute respiratory failure following years of slowly progressive pulmonary deterioration. His lung function was characterized first by mild airflow obstruction and later by progressive loss of lung volume. Computed tomography showed progressive development of pulmonary fibrosis and honeycombing. His serum disclosed precipitins to pigeon antigen. During his final illness his chest radiograph showed widespread patchy consolidation. At autopsy, his lungs revealed left lower lobe bronchopneumonia, fibrosis and honeycombing at the bases and widespread evidence of diffuse alveolar damage with organized exudate in some alveoli. To our knowledge, this is the second reported fatality due to acute alveolar injury in bird fanciers' lung.

Acute Disease↗

PubMed1997

Establishment of three monoclonal antibodies specific for prespermatogonia and intratubular malignant germ cells in humans.

Intratubular malignant germ cells (ITMGC), as assessed by clinicopathologic or cytogenetic studies, are regarded as a preinvasive lesion of all human testicular germ cell tumors with the exception of yolk sac tumors (in infants) and spermatocytic seminomas. To characterize specific surface molecules of ITMGC, we raised three mouse monoclonal antibodies (mAb) against NCR-G3 (G3), a multipotent, human embryonal carcinoma (EC) cell line, and screened cryostat sections of human testicular tissue containing ITMGC. These three mAb (HB5, IgG1; HF2, IgG1; HE11, IgG1) reacted to the surface of ITMGC, seminomas, and EC in vivo as well as to human EC cell lines in vitro. Expression of HB5 and HF2 antigens was down-regulated during cellular differentiation of G3 cells by retinoic acid or N,N'-hexamethylene-bis-acetamide treatment, whereas that of HE11 antigen was up-regulated with cellular differentiation by retinoic acid. Furthermore, these three mAb reacted to stage-specific prespermatogonia in the human fetus but not in human adults. HB5, HF2, and HE11 antigens were shown to be glycoproteins with molecular weights of approximately 80, 80, and 70 kd, respectively, and could be immunoprecipitated after deglycosylation treatment. Peptide mapping with Staphylococcus aureus V8 protease suggested that the HB5 and HF2 antigens were identical. We concluded that HB5/HF2 and HE11 antigens are oncodevelopmental antigens in testicular germ cell tumors and human spermatogenesis that may play a significant role in tumorigenesis and the development of human germ cells.

Adult↗

PubMed1996

A method for direct DNA amplification of uncharacterized DNA viruses and for development of a viral polymerase chain reaction assay: application to the red sea bream iridovirus.

A method is described for isolating a DNA segment of a virus for which no protein or DNA sequence information is available. This segment can then be used to develop a PCR-based assay for the virus. The method is based on the widespread presence and strong conservation of the ribonucleotide reductase gene among DNA viruses. The validity of the procedure is demonstrated by development of an assay for the fish iridovirus. We report the direct isolation from infected fish of a 738-bp segment of the iridovirus ribonucleotide reductase small subunit gene without prior virus purification. Using the sequence information obtained, a PCR-based diagnostic system was developed for detecting iridovirus infection.

Amino Acid Sequence↗

PubMed1996

Somatic mutations of the APC gene in sporadic hepatoblastomas.

Hepatoblastoma is a rare hepatic malignancy that occurs in children with an average age of 2 or 3 years and is known to be one of the extracolonic manifestations of familial adenomatous polyposis. Only a single hepatoblastoma with a germ-line mutation of the adenomatous polyposis coli (APC) gene has been reported thus far. To elucidate the possible roles of APC gene alterations in sporadic hepatoblastomas, we examined loss of heterozygosity (LOH) at the APC and MCC loci and performed a sequencing analysis of a part of the APC gene, including the mutation cluster region, in 13 hepatoblastomas of non-familial adenomatous polyposis patients. LOH at the APC and/or MCC loci was observed in four of seven (57%) informative cases. Of the 13 cases, somatic mutations were detected in 8 (61.5%), with 9 (69%) cases showing genetic alterations in the APC gene as LOH or somatic mutations. Two cases demonstrated double mutations. Furthermore, the nature of the somatic mutations observed in the present study was unusual because 9 of the 10 mutations were missense, with only 1 case featuring a frame-shift mutation due to an insertion. Previous reports have described almost all (>90%) mutations of the APC gene in colorectal tumors to result in a truncated APC protein due to either frame-shift or nonsense mutations. These findings suggest that a mutation of the APC gene may play an important role in the genesis of sporadic hepatoblastomas, and the mechanisms of APC gene alteration may be different from those reported previously for colorectal tumors.

Base Sequence↗

PubMed1996

Fish glucocorticoid receptor with splicing variants in the DNA binding domain.

Here we describe the isolation of a rainbow trout cDNA containing an entire GR coding region. Although the encoded protein is highly homologous to other GRs, especially in its DNA binding domain, it contains a nine amino acid insertion between the two zinc fingers. This novel form is found in all rainbow trout tissues examined; however, the testis also contains a splice variant lacking this insert, making it completely continuous to other GRs. In transient transfection assays of cultured cells, the two rainbow trout GR variants activated transcription from the glucocorticoid-responsive mouse mammary tumor virus promoter to comparable levels.

Alternative Splicing↗

PubMed1996

Comparative analysis of cell surface antigens expressed by cell lines derived from human germ cell tumours.

The pattern of cell surface antigen expression of a set of cell lines derived from human germ cell tumours and corresponding to various cell phenotypes found within these tumours was studied using immunofluorescence. Twenty-two different antibodies were used. Many of these antibodies have been noted to recognise epitopes that are either preferentially expressed by embryonal carcinoma (EC) cells, or by more differentiated cell types. Using scatter plots and rank correlations, 6 groups of antibodies were distinguished with respect to their staining patterns on the cell lines tested. Several antibodies showed a specific staining pattern in relation to the differentiation state of the cells. Two groups of antibodies included those recognising high m.w. glycoproteins (antibodies TRA-1-60, TRA-1-81, GCTM2, 3-177, K4 and K21) and the ganglioseries glycolipid antigens SSEA-3 and -4 (antibodies MC631 and MC813-70). These antibodies mostly stained EC cells but not other cell types, confirming previously published data. However, one of these groups, comprising antibodies K4 and MC631, was more exclusively associated with the EC cell phenotype than was the other group. Antibodies recognising the liver isozyme of alkaline phosphatase (TRA-2-49 and TRA-2-54) also reacted strongly with most EC cell lines, although they reacted significantly with a number of other cell lines as well, whereas antibodies to the placental isozyme tended to react only weakly with EC cells. The antibodies recognising the ganglioseries glycolipids GD2 and GD3 (VIN2PB22 and VINIS56) preferentially stained cells with neuroectodermal characteristics. Other antibodies showed a heterogeneous staining pattern for the cell lines with different phenotypes. The data obtained from the cell lines were, in general, similar to data obtained from immunohistochemical studies on tissue sections of primary germ cell tumours of the adult testis, including carcinoma in situ.

Adult↗

PubMed1996

Heat shock induces differentiation of human embryonal carcinoma cells into trophectoderm lineages.

NCR-G3 cells were established from a testicular embryonal carcinoma and are highly multipotential, differentiating into trophectoderm cells upon exposure to retinoic acid. Differentiated NCR-G3 cells begin to produce human chorionic gonadotropin (hCG), a trophectoderm-specific hormone. We have previously isolated the up-regulated genes at the early stage of differentiation. One of them was found to be a heat shock protein gene. The heat shock protein gene (HSP90) is induced at the early stage of differentiation and decreases to the basal level or under the basal level at the later stage. We speculate that heat shock per se induces the differentiation of human EC cells. With exposure to heat, NCR-G3 cells began to express a series of differentiation markers such as cytokeratin and hCG. Heat, which is classically known to induce heat shock proteins, is able to differentiate an embryonal cell line into trophectoderm lineages, implying a new recognized function of a heat-like event in early differentiation.

Carcinoma, Embryonal↗

PubMed1996

A novel chimera gene between EWS and E1A-F, encoding the adenovirus E1A enhancer-binding protein, in extraosseous Ewing's sarcoma.

Ewing's sarcoma/PNET, a tumor of the bone and soft tissue, is one of the most common causes of tumor death among youths. This tumor does not have specific phenotypes, but does have characteristic chromosomal translocations. Furthermore, the expression of EWS/FLI-1 or EWS/ERG chimeric genes was found to be generated through a t(11;22)(q24;q12) or a t(21;22)(q22;q12) translocation. In this study, we identified a new chimera gene between the transactivation domain of EWS and E1A-F, encoding the adenovirus E1A enhancer-binding protein. Since E1A-F is known to activate matrix metalloproteinase genes, the chimera gene may possibly be involved in tumor progression and could be a novel tumor marker for Ewing's sarcoma/PNET.

Adenovirus E1A Proteins↗

PubMed1996

Immunocytochemical localization of the Pit-1 protein in the pituitary of the rainbow trout (Oncorhynchus mykiss).

Pit-1 is a pituitary-specific transcription factor responsible for activating the growth hormone (GH) gene family. The localization of Pit-1 protein in the pituitary of rainbow trout was studied by immunocytochemical analysis. Fish Pit-1 was found to be extensively localized in the nuclei of cells in the proximal pars distalis (PPD), rostral pars distalis (RPD), and pars intermedia (PI). Sagittal sections were immunocytochemically double stained with antibodies against Pit-1, GH, prolactin (PRL), and somatolactin (SL). The results showed that Pit-1 protein was present in the GH-containing cells of the PPD, the PRL-containing cells of the RPD, and the SL-containing cells of the PI.

Animals↗

PubMed1996

Stage IVS neuroblastoma: a clinical, histological, and biological analysis of 45 cases.

This study reports clinicopathologic and biological analysis of 45 stage IVS neuroblastomas (IVS-NBs). All IVS-NB cases had small or undetectable primary tumors associated with disease involving the liver, skin, or bone marrow. Their ages at diagnosis ranged from 1 day to 12 months, with a median age of 4 months. The primary sites were adrenal in 34 cases, retroperitoneum in two, mediastinum in one, and unknown in eight; the liver was involved in 24 of 31 cases, the bone marrow in 11 of 27, and the skin in 4 of 29. According to the histopathological evaluation (Shimada classification), 43 IVS-NB tumors had a favorable histology, and 2 cases showed an unfavorable histology. Three patients died of progressive disease and six of indirect tumor death. Among these three deceased cases with tumor progression, two cases showed unfavorable histology, and one case had amplified N-myc (20 copies). These data suggest that most IVS-NB can be described as a tumor presenting characteristic dissemination pattern in the younger age group, having a favorable histology and showing an excellent outcome of the patients. However, poor prognosis group in stage IVS does exist, and this group could be detected histopathologically and biologically.

Adrenal Gland Neoplasms↗

PubMed1996

Mother to fry, successful transfer of immunity against infectious haematopoietic necrosis virus infection in rainbow trout.

We have tested whether immunity can be transferred from a mother fish to its fry. Rainbow trout mother fish were inoculated against infectious haematopoietic necrosis virus (IHNV) by intraperitoneal injection of a fragment of the IHNV glycoprotein spanning amino acids 31 to 310. This protein fragment was obtained by isolating the specific cDNA from Japanese IHNV strain HV7601 and expressing it in Escherichia coli. Fry from immunized and control fish were exposed to IHNV at various intervals after hatching, and their mortality monitored. Survival of the fry of immunized fish was significantly greater when exposure to virus occurred 7 days after hatching, and some immunity appeared to persist until at least 25 days after hatching.

Animals↗

PubMed1996

Expression of bone marrow stromal cell specific antigen during murine development: its expression in embryonic hematopoietic tissues as well as in other developing tissues.

Monoclonal antibody R4-A9 demonstrated specificity for a cell surface antigen of stromal cells in murine bone marrow and spleen. In order to identify patterns of expression that may elucidate the potential role of R4-A9 antigen, the developmental expression of this antigen in mouse embryos from 8 days post-coitum to 5 days post-partum was investigated by immunohistochemistry. At an early developmental stage, weak staining for R4-A9 antigen could be detected in the yolk sac. At later stages, strong staining of this antigen was detected predominantly in the embryonic liver, the main site of embryonic hematopoiesis. However, concomitant with the decreased staining in the liver, increased expression of this antigen was observed in bone marrow and spleen. Therefore, the changes in expression in those hematopoietic tissues suggest that its expression is coordinately regulated during the developmental stage of the sites of embryonic hematopoiesis. Compared with the distribution of R4-A9 antigen in adult tissues as previously reported, the expression of this antigen in fetal tissues was more widespread during the period of organogenesis, and was most abundant in other developing tissues, including the heart, skin, and lung. In contrast, fetal expression detected in hematopoietic and other developing tissues was lost after birth. These results taken together show a marked gradient of R4-A9 antigen expression, with the highest level at the peak of organ development, raising the possibility that this molecule may act as a growth/differentiation factor both in hematopoietic and other developing tissues in a fetus.

ADP-ribosyl Cyclase↗

PubMed1996

Identification of carriers of Duchenne/Becker muscular dystrophy by a novel method based on detection of junction fragments in the dystrophin gene.

We developed a Southern blotting based method that uses rare cutting restriction endonucleases and electrophoresis of single stranded DNA to detect junction fragments resulting from the rearranged dystrophin gene. By conventional Southern blot hybridisation, no junction fragments were detected in 27 unrelated patients with Duchenne (DMD) or Becker (BMD) muscular dystrophy, who had 20 deletions and seven duplications in the dystrophin gene. With our new method, junction fragments were detected in 21 of these 27 patients. When the junction fragments were used as markers, five carriers were unequivocally diagnosed among six females from two families of DMD/ BMD patients. This novel method allows simple and definitive identification of carriers with risk factors for DMD/BMD without using quantitative Southern blot hybridisation.

Blotting, Southern↗

PubMed1996

Induction of mcl1/EAT, Bcl-2 related gene, by retinoic acid or heat shock in the human embryonal carcinoma cells, NCR-G3.

NCR-G3 cells were established from a testicular embryonal carcinoma and were differentiated into multi-lineages including trophectoderm cells by exposure to retinoic acid. The differentiated cells began to produce human chorionic gonadotropin (hCG), a trophectoderm-specific hormone, which was regulated at the mRNA level. As we assumed that genes responsible for differentiation were differentially expressed at the early stage of retinoic acid-induced differentiation, we prepared a cDNA library from retinoic acid-treated NCR-G3 cells. This cDNA library was then screened for genes whose expression was induced during the differentiation of these cells. From about 5 x 10(4) clones screened, three independent sequences were isolated. Sequencing analysis revealed that clone 1002 codes for mcl1/EAT, which has a Bcl-2 homology domain. The expression of mcl1/EAT, the Bcl-2 related gene, was increased at an early stage of the retinoic acid-induced differentiation and preceded the up-regulation of cytokeratin and hCG genes after ratinoic acid treatment. Furthermore, mcl1/EAT was also up-regulated by heat shock, which has recently been shown to induce the cells to differentiate.

Biomarkers↗