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Biomedical subjects

S Noguchi

Publications and source records attributed to S Noguchi.

At least 343 records · Page 19Linked to original sources
PubMed1994

The detection of breast carcinoma micrometastases in axillary lymph nodes by means of reverse transcriptase-polymerase chain reaction.

BACKGROUND: The development of a sensitive method for the detection of breast carcinoma micrometastases in axillary lymph nodes is reported. METHODS: The method was based on amplification of MUC1 mRNA, which encodes a core protein of polymorphic epithelial mucin, by a reverse transcriptase-polymerase chain reaction (RT-PCR). Total RNA, which was extracted from a breast carcinoma cell line (MCF-7), primary breast carcinomas, and axillary lymph nodes, was subjected to analysis of MUC1 mRNA expression by the RT-PCR method. RESULTS: MUC1 mRNA expression was detected by RT-PCR in MCF-7 cells and in all 15 primary breast carcinomas but not in control lymph nodes taken from patients with benign diseases. A serial dilution study revealed that MUC1 RT-PCR was a very sensitive method, detecting one MCF-7 cell per 1,000,000 lymph node cells. The detection sensitivity of MUC1 RT-PCR method was compared with that of immunohistochemical staining of an epithelial marker (polymorphic epithelial mucin). Fifty axillary lymph nodes were obtained from 15 patients with primary breast carcinomas, and metastasis in each lymph node was investigated by both methods. The immunohistochemical method demonstrated metastasis in nine lymph nodes, and MUC1 mRNA was detected in all of them. Of the 41 lymph nodes that were diagnosed to be devoid of metastasis by immunohistochemistry, MUC1 mRNA was expressed by 6 but not by the other 35, indicating the presence of micrometastases in these 6 lymph nodes that could be detected only by the MUC1 RT-PCR method. CONCLUSIONS: The MUC1 RT-PCR method is more sensitive than immunohistochemistry for the detection of micrometastases in axillary lymph nodes. This new method would be of practical value in selecting the patients at high risk for relapse from those who are histologically lymph node negative.

Axilla↗
PubMed1994

Clonal analysis of human gynecologic cancers by means of the polymerase chain reaction.

Clonality of human gynecologic cancers was analyzed in small DNA samples prepared from cryostat sections, by means of the polymerase chain reaction (PCR). The method used for clonal analysis was based on restriction fragment length polymorphism of the X-chromosome-linked phosphoglycerokinase (PGK) gene and on the differential methylation of the PGK gene due to random inactivation of 1 of 2 X-chromosomes by methylation in females. Among 52 gynecologic cancers tested, 25 were found to be heterozygous for the BstXI polymorphism of the PGK gene. All the 25 gynecologic cancers (4 cervix, 11 endometrium, 7 ovary and 3 fallopian tube) analyzed by the PCR-based method were monoclonal in origin while adjacent normal tissues were polyclonal. When DNA samples were prepared from widely separated sites of tumors and/or metastatic lesions, every sample was found to be monoclonal, and the same allele of the PGK gene was inactivated in each case. These results demonstrate that clonal analysis by PCR offers a good method for studying clonality in small DNA samples prepared from cryostat sections of tumors. This method could be applied to distinguish between benign and malignant gynecologic lesions.

Adult↗
PubMed1994

Discrimination between multicentric and multifocal carcinomas of the breast through clonal analysis.

BACKGROUND: An unanswered, important question concerning multiple breast carcinomas is whether they arise independently in the breast (multicentric) or are metastatic deposits from a single, primary carcinoma (multifocal). This issue was studied by clonal analysis of each focus of multiple breast carcinomas. METHODS: First, the clonality of 30 breast carcinomas was analyzed by the method based on restriction fragment length polymorphism of the X-chromosome-linked phosphoglycerokinase (PGK) gene and on random inactivation of the gene. Second, the clonality of each focus of three multiple breast carcinomas was analyzed by the same method. RESULTS: Clonal analysis of the 30 breast carcinomas revealed that every carcinoma was monoclonal in origin, and one of two alleles of the PGK gene was inactivated at random in these carcinomas. Three patients with multiple breast carcinomas had three, three, and four histologically separate foci of carcinoma in the breast, respectively. Clonal analysis showed that each focus was monoclonal in origin and, in addition, the same allele of the PGK gene was inactivated consistently at each focus in every patient. CONCLUSIONS: The thesis that multiple carcinoma foci arise independently is unlikely to be true, because the probability that every independent focus happens to inactivate the same allele of PGK gene in every patient is very low. It seems more likely that a single primary carcinoma spreads throughout the breast to culminate in multiple secondary carcinoma foci. Therefore, it is concluded that multiple breast carcinomas are multifocal and not multicentric in origin.

Breast Neoplasms↗
PubMed1994

Simultaneous Assay of ErbB-2 Protein and Carcinoembryonic Antigen in Cyst Fluid as an Aid in Diagnosing Cystic Lesions of the Breast.

C-erbB-2 oncoprotein (ErbB-2 protein)and carcinoembryonic antigen (CEA)were simultaneously evaluated in the breast cyst fluid of 6 patients with intracystic cancer, 6 patients with intracystic papilloma, and 42 patients with gross cystic disease. A combination test, using 12 ng of ErbB-2 protein and 150 ng of CEA/ml of cyst fluid, respectively, as cutoff values resulted in a positive detection rate of 83%(5/6 cases) for intracystic breast cancer. Two of the 6 patients with intracystic papilloma and 2 of the 42 patients with gross cystic disease were positive. In addition, elevated levels of these markers in cyst fluid correlated well with the expression of these antigens in the corresponding tumor. Thus, it can be concluded that measurements of ErbB-2 protein and CEA aid in the diagnosis of intracystic breast cancer, and the assay of ErbB-2 protein is valuable for detecting tumors in which this protein is overexpressed.

Journal Article↗
PubMed1994

Preoperative Discrimination of Fibroadenoma Which Is Clinically and Cytologically Indistinguishable from Breast Carcinoma through Clonal Analysis of Fine Needle Aspirate of Tumor: Report of a Case.

Recently, it has been demonstrated that breast carcimoma is monoclonal and fibroadenoma is polyclonal in origin. In the present case report, this observation was successfully applied to a preoperative differential diagnosis of a fibroadenoma clinically and cytologically indistinguishable from carcimoma. Case report: A45-year old female presented for clinical examination with a history of breast lump. A firm tumor measuring 2 x 2 cm was palpable in the upper-outer quadrant of @the left breast. The margin of the tumor was partially ill-defined and its mobility was restricted. A tumor shadow with a partially ill-defined margin was revealed by mammography. Ultrasonographic examination showed and irregularly-shaped, hypo-echoic tumor accompanied by an acoustic shadow. Fine needle aspiration biopsy yielded positive cytology. Based on these results, the tumor was diagnosed as breast carcinoma. However, clonal analysis of fine needle aspirates showed a polyclonal pattern, strongly indicating that the tumor was not a carcinoma but benign disease; most probably fibroadenoma. Thus, an open surgical biopsy was performed. Histoglogical examination revealed that the tumor was indeed a fibroadenoma with epithelial hyperplasia. This case suggests the usefulness of clonal analysis of fine needle aspirates in differentiating fibroadenoma from carcinoma of the breast.

Journal Article↗
PubMed1994

[Granulocyte colony-stimulating factor treatment (G-CSF) of antithyroid drug-induced granulocytopenia: granulocyte count measurement after 4 hours of G-CSF injection is useful for the detection of recovery from granulocytopenia].

The primary objective of this study was to ascertain the usefulness of granulocyte count measurement after 4 hours of granulocyte colony-stimulating factor (G-CSF) injections for the detection of recovery from granulocytopenia. Four Graves' patients with antithyroid drug-induced granulocytopenia (granulocyte count between 500 and 1000/mm3) and three Graves' patients with antithyroid drug-induced agranulocytosis (granulocyte count < 500/mm3) each received a daily dose of 75 mu g of G-CSF administered subcutaneously. In all granulocytopenic patients, after 4 hours of G-CSF injection the granulocyte counts increased to 5623, 4050, 8923 and 4647/mm3, and the granulocyte count after 24 hours of G-CSF injection was 3008, 4634, 4854, 4200/mm3. In one of the three agranulocytic patients, the granulocyte count increased from 238/mm3 to 5982/mm3 after 4 hours of G-CSF injection, and the granulocyte count after 24 hours of G-CSF injection was 4800/mm3. Although the granulocyte counts before G-CSF injection of the remaining two agranulocytic patients were 138 and 126/mm3, the granulocyte counts after 4 hours of G-CSF injection were 837 and 59/mm3 and those after 24 hours of G-CSF injection were 817 and 0/mm3. These results indicated that granulocyte count measurement after 4 hours of G-CSF injection was useful for detecting the recovery from granulocytopenia and agranulocytosis.

Adolescent↗
PubMed1994

Dissociation of the complex of dystrophin and its associated proteins into several unique groups by n-octyl beta-D-glucoside.

Dystrophin is purified as a complex with several proteins from the digitonin-solubilized muscle cell membrane. Most of dystrophin-associated proteins (DAPs) are assumed to form a large oligomeric transmembranous glycoprotein complex on the sarcolemma and link dystrophin with a basement membrane protein, laminin. In the present study, we found that the purified dystrophin-DAP complex was dissociated into several groups by n-octyl-beta-D-glucoside treatment. In particular, we found that the glycoprotein complex stated above was dissociated into two distinct groups: one composed of 156DAG and 43DAG (A3a) and the other composed of 50DAG, 35DAG and A3b. We confirmed by crosslinking and immunoaffinity chromatography that these two groups existed in a complexes. We thus concluded that the glycoprotein complex consists of these two subcomplexes. Furthermore, A3b and 43DAG, which had been formerly treated simply as the 43DAG doublets due to their similar electrophoretic mobilities in SDS/PAGE, were shown to be present in two different subcomplexes. Based on the analyses by two-dimensional gel electrophoresis, peptide mapping and immunoblotting, we concluded that A3b is a novel DAP different from 43DAG.

Amino Acid Sequence↗
PubMed1994

Clonal analysis of parathyroid adenomas by means of the polymerase chain reaction.

Clonality of parathyroid adenomas and normal parathyroid glands was analyzed by a method based on restriction fragment length polymorphism of the X-chromosome-linked phosphoglycerokinase (PGK) gene and on random inactivation of the gene by methylation. Through the introduction of the polymerase chain reaction to this method, clonal analysis could be performed on small DNA samples prepared from cryostat sections of these specimens. Every normal parathyroid gland was found to be polyclonal while every parathyroid adenoma was found to be monoclonal. When DNA samples obtained from four widely separated sites of an adenoma were independently analyzed, each sample was found to be monoclonal and, in addition, the same allele of PGK gene was inactivated. These results suggest that parathyroid adenoma, which has a single cell origin, is a true neoplasm and that its pathogenesis is probably different from that of parathyroid hyperplasia which is polyclonal in origin.

Adenoma↗
PubMed1994

Clonal analysis of predominantly intraductal carcinoma and precancerous lesions of the breast by means of polymerase chain reaction.

Clonality of predominantly intraductal carcinoma (PIC) and precancerous lesions of the breast was analyzed by a method based on restriction fragment length polymorphism of the X-chromosome-linked phosphoglycerokinase gene and on random inactivation of the gene by methylation. The application of polymerase chain reaction to this method enabled clonal analysis of small lesions. In order to eliminate the contamination by normal stromal cells, intraductal components were microdissected from the frozen sections of PIC under a dissection microscope. Clonal analysis of the intraductal components from seven PICs revealed that all were monoclonal in origin. In three PICs with intraductal spreading of carcinoma cells over nearly a whole breast gland, the intraductal components were collected from eight widely separated sites in each case. Clonal analysis of these samples showed that every sample was monoclonal and the same allele of the phosphoglycerokinase gene was consistently inactivated in each case. These results suggest that PIC arises as a single monoclonal carcinoma and spreads through the ducts over the gland rather than having multicentric origins. Clonality of precancerous lesions such as atypical ductal hyperplasia and intraductal papilloma arising in the terminal ducts was also studied. Intraductal components were microdissected from the paraffin sections of these lesions and subjected to clonal analysis. Both atypical ductal hyperplasia and intraductal papilloma were found to be monoclonal in origin, suggesting that certain genetic changes had already occurred in the precancerous lesions. A further study is needed to elucidate these genetic changes, which would greatly help our understanding of the mechanism of carcinogenesis.

Base Sequence↗
PubMed1994

The functional roles of disulfide bonds in the beta-subunit of (Na,K)ATPase as studied by site-directed mutagenesis.

The beta-subunit of Torpedo californica (Na,K)ATPase contains seven cysteine residues; one (Cys46) is in the single transmembrane segment and the other six (Cys127, Cys150, Cys160, Cys176, Cys215 and Cys278) are in the extracellular domain and form three highly conserved disulfide bonds. A beta-subunit mutant with replacement of Cys46 by Ser could assemble with the alpha-subunit, and the resulting alpha beta-complex was catalytically active. Mutants in which either the N-terminal side or both Cys residues of the Cys127-Cys150 bond were replaced by Ser could also tightly assemble with the alpha-subunit, but the resulting alpha beta-complex was catalytically inactive. On the other hand, disruption of either the Cys160-Cys176 or Cys215-Cys278 bond by substituting the N-terminal side only or both Cys residues with Ser led to a beta-subunit that could not assemble with the alpha-subunit. We conclude that the structure of the beta-subunit around the Cys160-Cys176 and Cys215-Cys278 loops is indispensable for assembly with the alpha-subunit, whereas the Cys127-Cys150 loop is not essential for assembly but is required for enzyme activity.

Amino Acid Sequence↗
PubMed1994

Proton pumping activity and visible absorption and resonance Raman spectra of a cao-type cytochrome c oxidase isolated from the thermophilic bacterium Bacillus PS3.

Cytochrome c oxidase having heme O in addition to heme C and heme A (cytochrome cao) [Sone, N., & Fujiwara, Y. (1991) FEBS Lett. 288, 154-158] was isolated from a thermophilic bacterium, Bacillus PS3, grown under slightly air-limited conditions. Cytochrome cao could oxidize yeast cytochrome c and N,N,N',N'-tetramethyl-p-phenylenediamine twice as fast as cytochrome caa3, which this organism yielded under normal growing conditions. Cytochrome cao also pumped protons upon cytochrome c oxidation in a way similar to cytochrome caa3. Binding of cyanide to cytochrome cao caused spin-state conversion of heme O at the binuclear center and seriously inhibited its physiological activity. A low Ki value (0.4 microM) for cyanide was found to be mainly due to a small "off" constant. Resonance Raman spectra of cytochrome cao bore close resemblance to those of cytochrome caa3 in both oxidized and reduced states, although the formyl stretching (vCH=O) band was absent. The Fe-histidine stretching (vFe-His) and Fe-CO stretching (vFe-CO) frequencies of cytochrome cao were very close to those seen for cytochrome caa3, but were distinct from those of hemoglobin and peroxidases, suggesting that the protein structure in the vicinity of heme O resembles that of the heme a3 moiety of cytochrome caa3.

Bacillus↗
PubMed1994

Hemiatrophy of the tongue due to hypoglossal schwannoma shown by MRI.

Schwannomas account for 8.5% of all intracranial tumours; more than 90% arise from the 8th cranial nerve. Only 42 cases of schwannoma of the hypoglossal nerve have been reported. A 59 year-old woman developed right hemiatrophy of the tongue, clearly demonstrated on MRI, as was a small hypoglossal schwannoma. High signal was seen in the atrophic side of the tongue on both T1- and T2-weighted images, as described in the literature.

Cranial Nerve Neoplasms↗
PubMed1994

A phase II study of prophylactic intravesical chemotherapy with 4'-epirubicin in recurrent superficial bladder cancer: comparison of 4'-epirubicin and adriamycin.

Since intravesical recurrence of superficial bladder cancer (Ta, T1) after transurethral resection (TUR) is frequent, adjuvant therapy to reduce the recurrence rate has been extensively investigated. Although intravesical chemotherapy has been employed for 30 years or more, neither the exact effect on the bladder epithelium nor the optimal dose and administration schedule has yet been clarified. In recent years, several derivatives of Adriamycin (ADR) have been developed, and 4'-epirubicin (FARM) is one of them. This drug has been shown to have antitumor effects almost equal to those of ADR and to produce less toxicity when given systemically as chemotherapy. In an attempt to clarify the effect of intravesical FARM in the prevention of recurrence of superficial bladder cancer, we conducted a prospective randomized trial to compare the effects of equal doses of FARM and ADR given by intravesical instillation after TUR in cases of highly recurrent superficial bladder cancer. A total of 73 patients with recurrent superficial bladder cancer were randomized to receive TUR and either 30 mg FARM or 30 mg ADR by intravesical instillation every 2-4 weeks for 1 year. The prophylactic effect on recurrence and the toxic effects of these drugs were investigated. The current results show that FARM provides efficacy almost equal to that of ADR in the prevention of recurrence in these patients. However, FARM also caused almost the same local toxic effects (bladder irritation, among others) as ADR. On the basis of these preliminary results, FARM is surmised to be one of the agents as beneficial as ADR in the prevention of recurrence of superficial bladder cancer.

Administration, Intravesical↗
PubMed1994

Generalized resistance to thyroid hormone: identification of a novel c-erbA beta thyroid hormone receptor variant (Leu450) in a Japanese family and analysis of its secondary structure by the Chou and Fasman method.

Generalized resistance to thyroid hormone (GRTH) is characterized by elevated circulating levels of thyroid hormone in the presence of a eumetabolic state and failure to respond to triiodothyronine. Various point mutations in the c-erbA beta thyroid hormone receptor gene are known to be responsible for different phenotypes of GRTH. We herein report a new c-erbA beta variant in a Japanese family. The variant consisting of a cytosine to adenine base substitution at nucleotide position 1650 altered phenylalanine to leucine in codon 450 in the T3-binding domain of c-erbA beta. This base substitution was found in one allele of the 2 affected members of the family. The in vitro translation products of this mutant c-erbA beta gene demonstrated a significantly reduced T3-binding affinity. The secondary structure of this mutant thyroid hormone receptor predicted by the Chou and Fasman method included a new turn in the alpha helix structure in the T3-binding domain. We also discuss the secondary structures of the previously reported mutant receptors.

Adolescent↗
PubMed1994

Clonal analysis of regenerative nodules in hepatitis C virus-induced liver cirrhosis.

BACKGROUND/AIMS: Based on histological criteria, regenerative nodules in cirrhotic liver have been generally considered to result from hyperplastic proliferation of hepatocytes. Whether these nodules are hyperplastic or neoplastic has not been determined definitively. This study examined the issue by clonal analysis of each nodule. METHODS: The method for clonal analysis was based on restriction fragment length polymorphism of the X chromosome-linked phosphoglycerokinase gene and on random inactivation of the gene by methylation. RESULTS: Clonality of hepatocellular carcinoma (n = 7) and regenerative nodules (n = 76) induced by hepatitis C virus infection was analyzed. All carcinomas were monoclonal. Clonal analysis of regenerative nodules showed that 43% (33 of 76) were monoclonal in origin. Adjacent monoclonal nodules showed inactivation of the same allele of phosphoglycerokinase gene. Because the gene allele is inactivated at random, it is unlikely that each nodule happens to inactivate the same allele; it is more likely that monoclonal cell expansion is initiated before the nodule is established by septum formation. CONCLUSIONS: Monoclonal cell expansion is seen in a considerable number of regenerative nodules in cirrhotic liver. The results indicate that certain genetic changes, which are required for hepatocarcinogenesis, have already occurred in these nodules.

Base Sequence↗
PubMed1994

Spontaneous rupture of adrenal pheochromocytoma: a case report.

We report a case of retroperitoneal hemorrhage due to spontaneous rupture of a right adrenal pheochromocytoma, presenting as an acute abdominal emergency with symptoms of peripheral vasoconstriction. An elective operation was successfully performed on day 7 after sufficient volume replacement with continuous administration of an alpha and beta-adrenergic blocking agent.

Adrenal Gland Neoplasms↗
PubMed1994

pH-sensitive dissociation and association of beta-N-acetylhexosaminidase from boar sperm acrosome.

beta-N-Acetylhexosaminidase (beta-Hex, EC, 3.2.1.52) was released from cauda epididymal boar sperm by treatment with ionophore A23187, indicating that this enzyme is localized in the acrosome. beta-Hex was extracted on a large scale, with 2% acetic acid containing 0.2% Brij 35, from washed ejaculated sperm. By gel filtration chromatography, beta-Hex was separated into a high-molecular-weight fraction (beta-Hex I) and a low-molecular-weight fraction (beta-Hex I). beta-Hex I, which is predominant under acidic conditions (pH 6.5), dissociated into beta-Hex II under alkaline conditions (pH 7.4). beta-Hex II, converted from beta-Hex I, associated again to form beta-Hex I under acidic conditions. By sequential chromatography on ion-exchange, lectin, gel filtration, and ion-exchange HPLC columns, beta-Hex I and II were purified 1200-fold and 4000-fold, respectively, with a combined recovery of 23% as measured with synthetic substrate. An inhibitor of beta-Hex, O-(2-acetamido-2-deoxy-D-glucopyranosylidene) amino N-phenyl-carbamate (PUGNAC), reduced the in vitro fertilization rate in porcine cumulus-enclosed eggs, but barely changed the rate when cumulus-free eggs were used. beta-Hex I was shown to possess cumulus dispersion activity, suggesting that beta-Hex plays a role in the passing by sperm through cumulus cells before they bind to the zona pellucida.

Acetylglucosamine↗