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Biomedical subjects

M Miyagi

Publications and source records attributed to M Miyagi.

At least 19 recordsLinked to original sources

Correlation of loss of alleles on the short arms of chromosomes 11 and 17 with metastasis of primary breast cancer to lymph nodes.

To examine the role of loss of heterozygosity (LOH) during tumor development and/or progression, we looked for correlations between metastasis of breast cancer to a regional lymph node(s) and LOH of chromosomal arms 11p, 13q, 16q, 17p, and 17q, where frequent losses in primary tumors have been detected. No correlation between lymph node metastasis and LOH of chromosomes 13q, 16q, or 17q was observed. However, tumors showing LOH of chromosomes 11p (chi 2 = 10.82, P less than 0.01) and 17p (chi 2 = 6.78, P less than 0.01) revealed a significantly higher incidence of metastasis to a regional lymph node(s) than tumors without LOH on these chromosomal arms. Furthermore, only four of 30 (13%) patients with tumors that retained both 11p and 17p had metastasis to a regional lymph node(s), compared with 24 of 32 (75%) patients with tumors that had lost both 11p and 17p. Analysis of LOH with markers on chromosomes 11p and 17p in a large number of tumors indicated that the peritelomeric region of each of these chromosomal arms contains a tumor suppressor gene that may be associated with tumor progression, particularly metastasis to a regional lymph node(s).

Alleles

NH2-terminal acetylation of ribosomal proteins of Saccharomyces cerevisiae.

Using a mutant of Saccharomyces cerevisiae defective in the NAT1 gene, that encodes one of the NH2-terminal acetyltransferases, we have identified 14 ribosomal proteins whose electrophoretic mobility at pH 5.0 suggests they carry an additional charge, presumably due to the lack of NH2-terminal acetylation. At least 30 other ribosomal proteins from the mutant are electrophoretically normal. Attempted NH2-terminal analysis of most of the presumed acetylated proteins from wild type cells indicated that all were blocked. NH2-terminal analysis of the same proteins from the nat1 mutant strain yielded unique sequences. Each one carries an NH2-terminal serine. We conclude that these are normally acetylated due to the presence of the NAT1 gene product. It seems surprising that cells whose ribosomes have been altered to this degree grow rather well and synthesize the same spectrum of proteins as do wild type cells (Mullen, J. R., Kayne, P. S., Moerschell, R. P., Tsunasawa, S. Gribskov, M., Sherman, F., and Sternglanz, R. (1989) EMBO J. 8, 2067-2075). Finally, this analysis has provided the first sequence information available for several of the acetylated ribosomal proteins and for one non-acetylated ribosomal protein, which is clearly the product of the MFT1 gene (Garrett, J. M., Singh, K. K., Vonder Haar, R. A., and Emr. S. D. (1991) Mol. Gen. Gen. 225, 483-491).

Acetylation

Antibodies against 70-kD heat shock cognate protein inhibit mediated nuclear import of karyophilic proteins.

Previously, we found that anti-DDDED antibodies strongly inhibited in vivo nuclear transport of nuclear proteins and that these antibodies recognized a protein of 69 kD (p69) from rat liver nuclear envelopes that showed specific binding activities to the nuclear location sequences (NLSs) of nucleoplasmin and SV-40 large T-antigen. Here we identified this protein as the 70-kD heat shock cognate protein (hsc70) based on its mass, isoelectric point, cellular localization, and partial amino acid sequences. Competition studies indicated that the recombinant hsc70 expressed in Escherichia coli binds to transport competent SV-40 T-antigen NLS more strongly than to the point mutated transport incompetent mutant NLS. To investigate the possible involvement of hsc70 in nuclear transport, we examined the effect of anti-hsc70 rabbit antibodies on the nuclear accumulation of karyophilic proteins. When injected into the cytoplasm of tissue culture cells, anti-hsc70 strongly inhibited the nuclear import of nucleoplasmin, SV-40 T-antigen NLS bearing BSA and histone H1. In contrast, anti-hsc70 IgG did not prevent the diffusion of lysozyme or 17.4-kD FITC-dextran into the nuclei. After injection of these antibodies, cells continued RNA synthesis and were viable. These results indicate that hsc70 interacts with NLS-containing proteins in the cytoplasm before their nuclear import.

Amino Acid Sequence

Cloning and characterization of an interstitial deletion at chromosome 11p15 in a sporadic breast cancer.

In a number of types of cancer including breast, hepatocellular, and bladder carcinoma, frequent losses of heterozygosity (LOH) on chromosome 11p15 have indicated the presence of one or more tumor suppressor genes in this region. In the present study, we report the detection and characterization of a rearrangement at 11p15 in a sporadic breast carcinoma. Genomic DNA clones encompassing the rearranged region were isolated; localization of both flanking clones to 11p15 by two-color fluorescent in situ hybridization (FISH) indicated that the rearrangement was caused by an interstitial deletion in the affected allele. Although it is uncertain whether the region between the flanking two loci was missing from tumor cells, our result implied that a putative tumor suppressor gene on chromosome 11p15 is located between the loci on either side of the interstitial deletion or may be interrupted by one of the breakpoints.

Breast Neoplasms

Effects of sex hormones on chemotaxis of human peripheral polymorphonuclear leukocytes and monocytes.

The effects of sex hormones on the in vitro chemotaxis of polymorphonuclear leukocytes (PMNs) and monocytes were investigated using fMLP as the chemoattractant. PMNs, monocytes, and plasma were obtained from heparinized peripheral blood of healthy adults. Chemotaxis of PMNs or monocytes treated with sex hormones were tested using 48-well chemotaxis microchambers. The correlation between sex hormone levels in plasma and the chemotactic ability of PMNs from the same donor was also investigated. The chemotaxis of PMNs was enhanced by progesterone, while it was reduced by estradiol. Random migration of PMNs was also enhanced by progesterone and reduced by estradiol. The effect of estradiol on PMN chemotaxis was inhibited by addition of antiestrogens or progesterone. Testosterone did not have a measurable effect on PMN chemotaxis. A significant positive correlation was found between the concentration of progesterone in plasma of females and PMN chemotactic ability in vitro. For males, there was no significant relationship between plasma levels of sex hormones and PMN chemotactic ability. Estradiol and testosterone levels in plasma did not correlate with PMN chemotactic ability. Sex hormones had no effect on the chemotaxis of monocytes. These results suggest that the altered PMN chemotaxis associated with gingival inflammation may be due to the effects of sex hormones.

Adult

[Chemosensitivity of head and neck squamous cell carcinomas using subrenal capsule assay--comparative assay of 5-FU, FT, UFT, and HCFU and immunohistochemical study].

The chemosensitivity of 23 human head and neck squamous cell carcinomas (9 pharyngeal, 5 laryngeal, 5 oral cavity, 2 maxillary sinus, 1 salivary gland, 1 cervical esophagus) were examined using subrenal capsule (SRC) assays against 5-FU, FT, UFT and HCFU. Assays adequate for evaluation were obtained in 19 of 23 cases and the averages of tumor size changes on day 6 were, 17.3% for control, -0.8% for 5-FU, 10.3% for FT, -10.4% for UFT and 2.4% for HCFU. Sensitivity rates for each drug were 26.3% for 5-FU, 6.3% for FT, 38.9% for UFT and 11.1% for HCFU. To evaluate the clinical utility of these anticancer drugs, we determined the therapeutic index (TI), the ratio between tumor chemosensitivity and changes in animal body weight as a criterion for toxicity. The mean values of TI were 0.891 for control, 0.989 for 5-FU, 0.944 for FT, 1.054 for UFT and 0.980 for HCFU. UFT showed a greater antineoplastic effect as compared to the other three agents. Immunohistochemical study of the transplanted tumors revealed a specific antigen staining pattern similar to that observed in the original human cancer tissues. Degeneration of tumor tissues in drug-treated animals was also confirmed by immunohistochemical staining. Based on the results obtained from subrenal capsule assays, either by examining tumor size changes or evaluating TI, UFT proved to be the most effective of the 5-FU analogues against human head and neck squamous cell carcinomas.

Animals

[YAG laser contact therapy of ovarian endometriosis under laparoscopy].

Thirty-six patients with ovarian endometriosis were treated with Nd:YAG laser contact irradiation under laparoscopic control. Indications for laparoscopy were infertility (n = 20) and dysmenorrhea (n = 16). The laparoscopic procedures so far undertaken in our clinic include: Aspiration of chocolate cyst, removal of ovarian endometriosis, adhesion-lysis, uterine nerve ablation, coagulation of peritoneal endometriosis and irrigation. No complications were seen. After undergoing this procedure, eight of 20 patients achieved pregnancy and 15 of 16 patients obtained pain relief. Serum CA125 levels were significantly decreased postoperatively. We confirmed that contact irradiation with a cone-shaped sapphire probe provides adequate incision and lysis at lower power levels, and that this method is an effective treatment for ovarian endometriosis.

Adult

[A case of bronchiolitis obliterans organizing pneumonia in a patient with rheumatoid arthritis].

A 54-year-old man with rheumatoid arthritis visited his general practitioner because of fever and cough. Chest X-ray showed an infiltrative shadow in the right lower field. Antibiotic treatment was not effective, and the specimens obtained by transbronchial lung biopsy was not diagnostic. The patient was transferred to our hospital for further examination and treatment. Previously he had been treated with prednisolone in the knee joint for rheumatoid arthritis. Open lung biopsy was performed. The specimen obtained showed bronchiolitis obliterans organizing pneumonia (BOOP) histologically. The patient recuperated and the chest X-ray shadow decreased with no therapy except the previous treatment with prednisolone.

Arthritis, Rheumatoid

[Two case reports of septic shock due to Vibrio vulnificus with liver cirrhosis].

We have recently experienced a case of Vibrio vulnificus septicemia which occurred in a patient with hepatic cirrhosis, and as we were able to give early antibiotic treatment, the patient survived. We would like to report this case here together with another case experienced 2 years ago. Case 1 was a 58-year-old male who was attending our hospital as an outpatient for hepatic cirrhosis. At 5:30 pm on August 8, 1987, he consumed abalone and giant clam and at 9 pm complained of high fever with shaking chills. He was admitted to our department as an emergency case. Cefoperazone was administered resulting in a decline of fever on the following day. During the course of treatment he fell transiently into pre-DIC, but due mainly to the administration of antibiotics his condition was subsided. Case 2 was a 53-year-old male who was under medical care in our hospital for grave hepatic cirrhosis. On October 11, 1985, he consumed sushi and two days later suffered chills and pyrexia. A blood culture revealed Vibrio vulnificus. His condition improved transiently with administration of Cefazolin, but oliguria, hypotension and ascites occurred subsequently, and finally the patient died on the 22nd day.

Humans

[Bacteriological study of gingivitis in pubertal children. 2. Subgingival microflora cultivable on the nonselective blood agar medium].

The present study involved cultural and microscopic characterization of the subgingival microorganisms of pubertal children with gingivitis. Samples were obtained from eight children 12-14 years of age. Gram-positive organisms made up 56% of the isolates and included mainly Actinomyces israelii, Actinomyces naeslundii. Gram-negative anaerobic rods constituted 25% of the isolates and Bacteroides intermedius (9.7%) was most predominant species. The selective media permitted significantly lower recovery of Actinomyces spp. and Capnocytophaga spp. than nonselective blood agar medium. However, sixty-nine percents of isolates were cultured on the selective media employed in the present study. These results suggest that the subgingival microflora of gingivitis in pubertal children may be similar to those of adult gingivitis and that the use of these six selective media makes it possible to enumerate predominant cultivable microorganisms in subgingival area of pubertal children with gingivitis.

Actinomycetales

Hypocholesterolemic activity of alpha-amylase hydrolysate of bagasse alkaline extract in rats fed atherogenic diet.

Cholesterol-lowering activity of enzymatic hydrolysate of bagasse alkaline extract was studied in rats fed a cholesterol-enriched diet. The bagasse alkaline extract was found to be vulnerable to the enzymic attack of alpha-amylase. Thus the alkaline extract was degraded into two fractions: long-chain fraction (polysaccharides) and short-chain fraction (oligosaccharides). The long-chain fraction was active in lowering the serum cholesterol level while feeding the short-chain fraction resulted in a slightly elevated serum cholesterol level. The feeding by gavege of the long-chain fraction 12 h after the meal also was effective in lowering serum cholesterol concentration. These findings were compatible with the concept that the bagasse alkaline extract affect the cholesterol dynamics apart from input-output balance of cholesterol. Possible mechanisms responsible for the cholesterol-lowering activity of the extract were discussed.

Amylases

[Gastric acid secretions and serum gastrin levels in patients with mucosal and submucosal gastric cancers].

Gastric acid secretions and serum gastrin levels have been examined in 128 patients with early gastric cancer and in 98 gastric ulcer patients. Gastric cancer patients were found to have lower acid secretions than did gastric ulcer patients, and those with elevated types of a differentiated adenocarcinoma had lower acid secretions than did those with depressed types of an undifferentiated adenocarcinoma. Gastric acid secretions in patients with both a gastric ulcer and cancer were found to decrease with aging. However, the serum gastrin levels were found to be decreased in patients with a gastric ulcer and to be increased in patients with a gastric cancer. Incidences of a differentiated adenocarcinoma increased with aging. From these observations, it has been speculated that the carcinogenesis of a differentiated adenocarcinoma may be related to increasing endogenous gastrin levels and decreasing gastric acid secretions. These results suggest that a continuous check of the serum gastrin levels might be a good marker for cancer detection and that gastrin antibodies might be useful for treatment.

Adenocarcinoma

Increased integration of viral genome following chemical and viral treatment of hamster embryo cells.

Treatment of hamster embryo cells with diverse classes of chemical carcinogens enhances transformation by a carcinogenic simian adenovirus, SA7. Virus transformed foci selected from plates pretreated with 3-methyl-cholanthrene (MCA), methyl methanesulfonate (MMS) or 7,12-dimethylbenz[a]anthracene (DMBA) and established as cell lines in culture, contained equivalent amounts of SA7 viral genome. However, hamster embryo cultures treated with MMS or nickel sulfate had increased amounts of SA7 DNA integrated into cellular DNA when examined 2--9 days after chemical treatment and viral inoculation. An increased uptake of SA7 DNA was demonstrated in hamster cells treated with MMS during DNA repair synthesis in cells retricted in scheduled DNA synthesis by amino acid deprivation; addition of virus after the repair period did not result in an increased integration of viral DNA. These data suggest that enhancement of viral oncogenesis by chemical carcinogens or mutagens may be related to the formation of additional attachment sites in cellular DNA for insertion of viral DNA, thereby increasing the probability of viral transformation.

9,10-Dimethyl-1,2-benzanthracene

Reverse transcriptase from avian myeloblastosis virus.

From lots of 20 to 30 g of avian myeloblastosis virus RNA-dependent DNA polymerase was obtained in preparations of purity greater than 95% by using a two-step column chromatographic procedure employing DEAE (DE 52) and carboxymethylcellulose (CM 52.). Yields of RNA-dependent DNA polymerase varied from approximately 20,000 to 35,000 U/g of virus. Specific activity of the enzyme was about 35,000 to 60,000 U/mg of protein. Free of detectable RNase activity, the product exhibited a molecular weight of about 160,000, an isoelectric point of 6.5, and approximately 2 mol of fatty acid per mol of enzyme.

Animals

Protein kinase from avian myeloblastosis virus.

A protein kinase associated with purified virions of avian myeloblastosis BAI strain A was partially purified by ion-exchange chromatography and gel filtration. The transfer of phosphate catalyzed by this enzyme required a divalent metal ion and ATP as phosphate donor. GTP could not be substituted for ATP, and the reaction was unaffected by either cyclic AMP or beef-heart protein-kinase inhibitor. Of the virus and nonvirus proteins tested as phosphate acceptors, only acidic proteins were phosphorylated. In particular, purified preparations of reverse transcriptase from avian myeloblastosis virus did not accept phosphate. The enzyme is a basic protein (pI = 9.3), and, on the basis of molecular sieving through Sephadex G-200 and velocity sedimentation on glycerol gradient, the protein kinase has a molecular weight of 45,000.

Adenosine Triphosphate