Biomedical subjects

Y Nakanishi

Publications and source records attributed to Y Nakanishi.

At least 19 recordsLinked to original sources

PubMed1992

A silencer-like cis element for the testis-specific phosphoglycerate-kinase-2-encoding gene.

Phosphoglycerate kinase (PGK), a glycolytic enzyme, possesses two isozymes: somatic-type PGK-1 and testis-specific PGK-2, encoded by distinct genes. Tissue-specific expression of the two PGK-encoding genes (Pgk) seems to be transcriptionally controlled, since tissue distribution of the mRNAs coincides well with that of the proteins. In the present study, we determined the cis-acting DNA elements that regulate the transcription of mouse Pgk-2. A transient expression assay of DNAs having various portions of the Pgk-2 upstream region linked to the chloramphenicol acetyltransferase (CAT)-encoding gene (cat) was performed using mouse cell lines that exclusively express Pgk-1. A substantial increase in cat expression was observed when the region between nucleotides (nt) -1404 and -685, relative to the most distal transcription start point at nt +1, was lost. This cis-acting region appeared to function as a silencer, since it repressed cat expression independently of either orientation to or distance from the Pgk-2 promoter. Moreover, the cis element inhibited Pgk-2 transcription with no effect on Pgk-1 transcription in a cell-free system using nuclear extracts of rat liver. These results suggest that a silencer-like negative cis element is responsible, at least partly, for tissue-specific transcription of Pgk-2.

Animals↗

PubMed1992

Direct effect of basic fibroblast growth factor on gene transcription in a cell-free system.

Recent findings on the translocation of intact fibroblast growth factor (FGF) into the cell nucleus suggest that it functions directly in nuclear events. We examined the effect of human basic FGF (bFGF) on gene transcription in a cell-free system. When mouse genes encoding phosphoglycerate kinases 1 and 2 (Pgk-1 and Pgk-2) were transcribed by using nuclear extracts of Ehrlich ascites tumor cells, FGF affected transcription in different ways: in the presence of bFGF, transcription of the Pgk-1 gene was inhibited, whereas that of the Pgk-2 gene was enhanced. When viral genes were tested, transcription of the adenovirus major late DNA was slightly stimulated but that of the adenovirus early E1A DNA or the human immunodeficiency virus DNA was not changed by the addition of bFGF. Moreover, the presence of a distinct 5' upstream region of the Pgk-2 gene, which includes a negative cis-acting element, was required for transcription stimulation by bFGF. These results suggest that bFGF can regulate transcription directly in the nucleus in a gene-specific manner.

Adenovirus Early Proteins↗

PubMed1992

[Clinical usefulness of 99mTc-PMT whole body scans in the diagnosis of bone metastases from hepatocellular carcinoma].

The diagnostic value of whole body scanning using 99mTc-N-pyridoxylmethyltryptophan (PMT) was evaluated in 16 patients with bone metastases from hepatocellular carcinoma, in comparison with 99mTc-MDP. Of the 72 known lesions of bone metastases, 63 (87.5%) were detected by 99mTc-PMT scintigraphy, which demonstrated increased uptake of radionuclide. However, 99mTc-MDP bone scintigraphy detected only 45 lesions (62.5%), which were shown as increased, decreased, or mixed patterns of uptake. Thus 99mTc-PMT scintigraphy was more sensitive than 99mTc-MDP bone scintigraphy. In addition, the latter showed poor specificity because of its high false positive rate due to degenerative change. All lesions undetected by 99mTc-PMT scintigraphy were located in areas that overlapped the liver or bowel activity. In conclusion, it is recommended that whole body 99mTc-PMT scintigraphy be combined with 99mTc-MDP bone scintigraphy for the detection of bone metastases from hepatocellular carcinoma.

Aged↗

PubMed1992

Isolation of new temperature-sensitive mutants of Saccharomyces cerevisiae deficient in mannose outer chain elongation.

We have isolated two temperature-sensitive Saccharomyces cerevisiae mutants which exhibit a deficiency in mannose outer chain elongation of asparagine-linked oligosaccharide. The size of yeast glycoprotein, secretory form of invertase, of one mutant (och1) was slightly larger than that of the sec18 mutant at the non-permissive temperature, while that of the other mutant (och2) was almost the same as that of the sec18 mutant. Unlike sec mutants, the och mutants were not deficient in secretion of invertase. The och1 mutant showed a 2+:2- cosegregation with regard to the temperature sensitivity and mannose outer chain deficiency, suggesting that a single gene designated as OCH1 is responsible for these two phenotypes. The och1 mutant stopped its growth at the early stage of bud formation and rapidly lost its viability at the non-permissive temperature. The och1 mutation was mapped near the ole1 on the left arm of chromosome VII. The och1 mutant cells accumulated the external invertase containing a large amount of core-like oligosaccharides (Man9-10GlcNAc2) and a small amount of high mannose oligosaccharides (greater than Man50GlcNAc2) at the non-permissive temperature. Production of the active form of human tissue-type plasminogen activator was increased in the och1 mutant compared with the parental strain, suggesting the potential advantage of this mutant for the production of mammalian-type glycoproteins which lack mannose outer chains in yeast.

Asparagine↗

PubMed1992

Multiple cis-acting DNA elements that regulate transcription of the adenovirus 12 E1A gene.

To delineate cis-acting elements for adenovirus (Ad) 12 E1A gene transcription, we transfected HeLa and NIH3T3 cells with DNAs having various deletions in the 5'-upstream region linked to the chloramphenicol acetyltransferase gene. Deletions in the regions between nucleotide (nt) positions 54 and 166, and 167 and 200, with respect to the left end of the viral genome at nt position 1, caused a two- to three-fold reduction in transcription. Transcription decreased to an almost undetectable level with loss of the region between nt positions 201 and 282. The effect of these mutations was almost consistent between both cell lines. The region between nt positions 77 and 94 stimulated transcription when situated upstream of the simian virus 40 early promoter in either orientation. Transcription was stimulated about ninefold in the presence of the DNA that encodes the product of the 13S, but not the 12S mRNA of the Ad12 E1A gene. These results indicate that transcription of the Ad12 E1A gene is regulated by multiple cis-acting elements and is stimulated by its own gene product.

3T3 Cells↗

PubMed1992

A vascularised rib strut technique for funnel chest correction.

We have refined the Ravitch technique of sternal elevation for surgical correction of funnel chest. Our major modification is introducing a living rib strut for supporting the elevated sternum. The left 5th rib with a vascular pedicle from the internal thoracic artery is turned 180 degrees beneath the sternum. We have used this method in 10 cases. The results have all been satisfactory.

Adolescent↗

PubMed1992

Co-culture of in vitro fertilized bovine embryos with different cell monolayers.

The effects of different cell monolayers on in vitro development of early bovine embryos derived from in vitro maturation and fertilization were examined in this study. Early embryos (four to eight cells) were randomly allocated to bovine granulosa cell (GC), oviductal cell (OC), or uterine cell (UC) monolayers in Exp. 1 and to GC, skin cell (SC; from 10-d-old chicken embryos), testicular cell (TC; from 10-d-old mouse), and liver cell (LC; from 10-d-old chicken embryos) monolayers in Exp. 2, and cultured for 6 d at 38.6 degrees C in a humidified atmosphere of 5% CO2 in air. The culture medium was 12.5 mM HEPES TCM 199 supplemented with 1% calf serum and 1 mM sodium pyruvate. In Exp. 1, the percentage of four- to eight-cell embryos that developed to blastocysts on GC, OC, and UC monolayers was 26.9 (28/104), 37.5 (39/104), and 39.2 (40/102), respectively. In Exp. 2, the percentage of four- to eight-cell embryos that developed to blastocysts on GC, SC, TC, and LC monolayers was 53.3 (40/75), 42.9 (33/77), 49.3 (37/75), and 44.3 (35/79), respectively. There were no significant differences in development among groups in either experiment.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1992

[Disappearance of lung metastases from hepatocellular carcinoma following bronchial arterial infusion of CDDP and MMC].

A 58-year-old man was admitted to our department because he had been diagnosed as hepatocellular carcinoma, which was located at S6 segment, and posterior segmentectomy was performed. After 6 months, right lung metastases of HCC were found and right bronchial arterial infusion of CDDP and MMC was performed twice. Dramatic effects were obtained such as disappearance of lung metastases. We emphasize the useful effect of CDDP and MMC for metastases of HCC.

Antineoplastic Combined Chemotherapy Protocols↗

PubMed1992

[Combination chemotherapy with 254-S, ifosfamide and peplomycin for advanced or recurrent cervical cancer].

From favorable results with 254-S, a new cisplatin analogue, single administration, we have conducted a clinical study to investigate the efficacy of combination of 254-S, ifosfamide and peplomycin, each of which has a different dose limiting factor. A total of 45 patients, including 22 patients with stage III and IV cervical cancer and 23 cases with recurrent cervical cancer, were treated with at least two courses of 254-S (100mg/m2, iv. Day 1), ifosfamide (1,500mg/body, iv. Day 1-5) and peplomycin (5mg/body, im. Day 1-6), and tumor response was evaluated clinically and by CT scanning. The response rate obtained in patients with advanced disease was 81.8% (PR = 17, CR = 1) and that in cases with recurrence was 60.9% (PR = 12, CR = 2). Myelosuppression was the dose limiting factor. In the 121 courses, grade 3 and 4 of leucopenia and thrombocytopenia were observed with an incidence of 44% and 32%, respectively and DIC occurred in 3 cases with poor PS though they recovered after reducing the 254-S dose to 80 mg/m2. The other toxicities were mild except for alopecia. Anaphylaxia was observed in a case at the second administration though the patient recovered in 15 minutes. There was no death. As to prognosis, a significant prolongation of survival period was observed in recurrent cases and 4 cases are alive (NED) after one and a half year. In the advanced cases, until now 3 cases of stage IV have died from the disease. We have concluded that this regimen is effective as a neoadjuvant chemotherapy for advanced cervical cancer and useful for the treatment of recurrent cervical cancer.

Adenocarcinoma↗

PubMed1992

Salivary gland morphogenesis: possible involvement of collagenase.

When 12-day rudiments of mouse submandibular glands were cultured, they formed an average two clefts within 24 h. The average number of the clefts, however, increased up to five in the presence of bovine tissue inhibitor of metalloproteinases (TIMP). Contrary to this, either bacterial or bovine interstitial collagenase in the medium completely inhibited the cleft formation of the glands. Electron microscopic observations revealed that the amounts of collagen bundles at the cleft points significantly increased with TIMP, but decreased with bacterial collagenase. These results strongly support the idea that endogenous collagenase regulates the cleft formation through the modulation of fibrillar architectures containing collagen in the extracellular matrix. Recently, our immunohistochemical studies clarified that collagen III, known to be rich in embryonic tissue, accumulated preferentially at the cleft points of the epithelium, and may play a crucial role in submandibular gland morphogenesis.

Animals↗

PubMed1992

[Neurological manifestations in hemorrhagic colitis associated with Escherichia coli O 157: H 7].

From September through November 1990, an outbreak of hemorrhagic colitis associated with Escherichia coli O 157: H 7, occurred in a kindergarten in Saitama, Japan. Some of the patients suffered from neurological symptoms such as stupor, deep coma and/or convulsions in the acute stage, and/or action tremors, nystagmus, incontinence, phrenic nerve palsy in the later stage. Serum complements decreased more in the patients with neurological symptoms than in the patients without them. The verotoxin elaborated by E. coli O 157: H 7 was considered to cause the neurological symptoms, on the basis of their clinical courses and laboratory findings, especially the cerebrospinal fluid findings.

Child↗

PubMed1991

Transcription stimulation of the adenovirus type-12 E1a gene in vitro by a novel factor bound to a region adjacent to a TATA box.

The E1a gene of adenovirus (Ad) type-12 possesses two transcription start points (tsp) separated by 139 nucleotides (nt). We previously found that transcription from a tsp distal to the coding region is preferentially stimulated in a cell-free reaction by nuclear factor I (NF-I) bound to a region near the left end of the virus genome. We report here on the identification of a cis-acting DNA region and a trans-acting factor for transcription initiated at the proximal tsp of the Ad12 E1a gene. A deletion in the region between nt -50 and -36 relative to the proximal tsp at +1 caused a significant decrease in the proximal transcription in a cell-free reaction using nuclear extracts of HeLa cells. A cellular factor binding to this region was shown to be responsible for transcription stimulation. This E1A-stimulating factor (ESF-1) appeared to recognize the sequence 5'-TGTCA-3' located immediately upstream from a TATA box. Unlike NF-I, the ESF-1 activity did not seem to be influenced by the E1A protein. Our results indicated that ESF-1 stimulates the proximal transcription of the Ad12 E1a gene by binding to the region adjacent to a TATA box.

Adenoviridae↗

PubMed1991

[Clinical application of asialoglycoprotein receptor-mediated liver scintigraphy using 99mTc-DTPA-galactosyl-human serum albumin].

Technetium-99m-diethylenetriaminepentaacetic acid-galactosyl-human serum albumin (99mTc-GSA) is a new liver scintigraphy agent which binds to asialoglycoprotein receptors on the hepatocytes. Studies were performed in three normal volunteers and 19 patients with chronic liver diseases. Serial scintigrams and time-activity curves of heart and lever were obtained for 60 min following a single intravenous injection of 99mTc-GSA (1 mg/185 MBq). % Injected dose (%ID) in the blood at 60 min after injection and that in the liver were determined. Excellent hepatic images were obtained for 60 min in normal subjects, and neither blood pool image nor extrahepatic uptake was recognized. In the studies of patients, however, blood pools were also visualized in various degrees suggesting altered receptor quantity, although the livers were visualized clearly. The %ID in the blood at 60 min after the injection showed significant correlations with serum albumin levels (r = -0.741, p less than 0.0001), retention rates of ICG at 15 min (r = 0.855, p less than 0.0001), plasma disappearance rates of ICG (r = -0.760, p less than 0.0005), prothrombin times (r = -0.696, p less than 0.002), hepaplastin tests (r = -0.575, p less than 0.05), total bilirubin levels (r = 0.689, p less than 0.001) and Child Turcotte scores (r = 0.769, p less than 0.0001). Significant differences of %ID in both blood and liver were observed between normal subjects and patients in accordance with severities of the hepatic disorders. These results suggest that 99mTc-GSA liver scintigraphy is useful in evaluating the liver function.

Adult↗

PubMed1991

Transcription inhibition of the somatic-type phosphoglycerate kinase 1 gene in vitro by a testis-specific factor that recognizes a sequence similar to the binding site for Ets oncoproteins.

To elucidate the mechanism by which transcription of the somatic-type phosphoglycerate kinase 1 gene is inactivated during mammalian spermatogenesis, we examined the presence of specific transcription inhibitor(s) in the testis by a cell-free transcription system. Transcription of the mouse phosphoglycerate kinase 1 gene using nuclear extracts of the rat liver was significantly inhibited by the addition of testis extracts, whereas brain extracts had little effect. Transcription inhibition required the binding of a testis-specific factor, designated TIN-1, to the region between positions -268 and -259 relative to transcription initiation site at +1. This region had the sequence 5'-AGGAAGTTCC-3' that includes an inverted repeat of the binding motif, 5'-GGAA-3', for the oncoprotein Ets. A UV-crosslinking experiment revealed that 43- and 45-kDa polypeptides present in testis extracts bind to that sequence. These results suggest that a testis-specific transcription inhibitor TIN-1 inactivates the phosphoglycerate kinase 1 gene in the mammalian spermatogenic pathway.

Animals↗

PubMed1991

Application of lectin and B-lymphocyte-specific monoclonal antibodies for the demonstration of human microglia in formalin-fixed, paraffin-embedded brain tissue.

To evaluate the usefulness of microglial markers for routine neuropathological material, we studied formalin-fixed, paraffin-embedded human brain tissue with the immunoperoxidase method using the lectin Ricinus communis agglutinin (RCA-1) and four monoclonal antibodies (LN-1, LN-2, LN-3, anti-HLA-DR/alpha). RCA-1 stained resting microglia, but the staining intensity was mostly weak. LN-1 also stained resting microglia in paraffin sections first treated with protease. In contrast to LN-1, RCA-1 stained blood vessels heavily. LN-1 stained resting microglia more markedly than RCA-1 in brains fixed for a prolonged period of time. However, LN-1 recognized a small number of astrocytes in routine paraffin sections. LN-3 reactivity was detected on a few resting microglia, but was intensely expressed on large numbers of reactive microglia in many neurological diseases. Both LN-2 and anti-HLA-DR/alpha labelled microglia, but the reactions were inconsistent. This study suggests that the monoclonal antibodies LN-1 and LN-3 are useful for the demonstration of microglia in paraffin sections, and a combination of these antibodies and the antibody to glial fibrillary acidic protein is recommended in attempting to identify microglia.

Antibodies, Monoclonal↗

PubMed1991

Comprehensive treatment of bilateral cleft lip by multidisciplinary team approach.

Repair of bilateral cleft lip presents numerous problems, and in our opinion, it is better to begin treatment at the earliest age possible. At Fujita Health University Hospital, we utilise a multidisciplinary team approach to cleft lip. Nonsurgical correction of the nasal deformity using a nose retainer and preoperative orthodontics using a Kuwahara-modified Hotz's palatal plate begins soon after birth. Surgical repair of the lip is done within the first 2 months of life, by the time the nose, alveolus and projecting prolabium are adequately reformed. A one-stage surgical procedure, including restoration of muscle union, labial sulcus construction and nasal correction is performed. After lip repair, lip and tongue pressure are well balanced by the plate, and a good alignment of the alveolus can be achieved. A total of 27 cases of bilateral cleft lip were treated from August 1986 to October 1990. In all cases, the postoperative course was uneventful, and no complications due to early surgery were encountered.

Cleft Lip↗

PubMed1991

Immunohistochemical localization of five classes of intermediate filament in a benign pelvic soft tissue tumor of rhabdoid appearance.

A benign pelvic soft tissue tumor from a 50-year-old woman was examined by immunohistochemistry and electron microscopy. The tumor cells had abundant eosinophilic cytoplasm with a hyaline appearance, which was filled with large aggregates of intermediate-sized filaments (IF). The cells were positively immunostained by antibodies against cytokeratin, vimentin, desmin, glial fibrillary acidic protein, and neurofilament proteins. This case represents an extreme example of the simultaneous expression of IF by neoplastic cells, and exemplifies the limited applicability of immunohistochemical detection of IF antigens for pathological diagnosis of neoplasms.

Female↗

PubMed1991

A novel cis-acting element that controls transcription of human immunodeficiency virus type 1 DNA, depending on cell type.

cis-acting elements for the transcription of human immunodeficiency virus type 1 DNA were analyzed in cell-free transcription and DNA transfection assays. Besides previously identified cis elements, a region adjacent to the enhancer element was found to regulate transcription in both assays. Loss of this region caused 4.3- and 1.6-fold transcription inhibition in a transfection assay with a T-cell line, MOLT-4, and a monocyte line, U937, respectively, whereas the same region appeared to function negatively with other T-cell lines, MT-4 and Jurkat. These results suggest that this novel cis element regulates the transcription of proviral DNA in a cell-type-specific manner.

Base Sequence↗