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Biomedical subjects

S Nomoto

Publications and source records attributed to S Nomoto.

At least 55 records · Page 3Linked to original sources

Immunohistochemical and molecular biological studies of serous cystadenoma of the pancreas.

Seven cases of pancreatic serous cystadenoma were examined immunohistochemically and molecular biologically. Six were benign tumors and one was clinically malignant. Immunohistochemical studies were performed with the avidin-biotin peroxidase complex technique on paraffin-embedded tumor tissue and were stained with antibody to carcinoembryonic antigen (CEA), CA19-9, and p53 protein. Two-stage polymerase chain reaction-restriction fragment length polymorphism analysis was used to detect K-ras oncogene mutation at codon 12. No tumor cells were stained with anti-CEA and anti-p53 protein, but two cases were stained focally with anti-CA19-9. One case was benign and one was clinically malignant. In the anti-CA19-9 staining, tumor cells of the benign case were positive only on the apical membrane and supranuclear cytoplasm of the cells, whereas those of the clinically malignant case were positive over the entire surface and cytoplasm of the cells. All seven cases were without K-ras gene mutation. So the features of serous cystadenoma of the pancreas suggest a tumor genesis different from that of ductal adenocarcinoma. They also suggest a relationship between immunohistochemical localizations of CA19-9 in the tumor cells and the biological behavior of the tumor itself.

Adult↗

Comparisons between in-vitro and in-vivo metabolism of (S)-warfarin: catalytic activities of cDNA-expressed CYP2C9, its Leu359 variant and their mixture versus unbound clearance in patients with the corresponding CYP2C9 genotypes.

To study whether an in-vitro model for three different genotypes of human CYP2C9*3 polymorphism would be useful for predicting the in-vivo kinetics of (S)-warfarin in patients with the corresponding genotypes, the intrinsic clearance (Cl(int) or Vmax/Km) for (S)-warfarin 7-hydroxylation obtained from recombinant human CYP2C9*1 [wild-type (wt)] and CYP2C9*3 (Leu359/Leu) expressed in yeast and the mixture of equal amounts of these were compared with the in-vivo unbound oral CI (CI(po,u)) of (S)-warfarin obtained from 47 Japanese cardiac patients with the corresponding CYP2C9 genotypes. The in-vitro study revealed that the recombinant CYP2C9*1 (wt/wt), 2C9*3 (Leu359/Leu) and their mixture (Ile359/Leu) possessed a mean Km of 2.6, 10.4 and 6.6 microM and Vmax of 280, 67 and 246 pmol/min/nmol P450, respectively. Thus, the mean in-vitro Cl(int) obtained from recombinant CYP2C9*3 (Leu359/Leu) and the mixture (Ile359/Leu) of 2C9*3 and 2C9*1 were 94% and 65% lower than that obtained from CYP2C9*1 (wt/wt) (6.7 versus 38 versus 108 ml/min/micromol P450, respectively). The in-vivo study showed that the median Cl(po,u) for (S)-warfarin obtained from patients with homozygous (Leu359/Leu, n = 1) and heterozygous (Ile359/Leu, n = 4) CYP2C9*3 mutations were reduced by 90% (62 ml/min) and 66% (212 ml/min, P < 0.05) compared with that obtained from those with homozygous 2C9*1 (625 ml/min, n = 42). Consequently, there was a significant correlation (r = 0.99, P < 0.05) between the in-vitro Cl(int) for (S)-warfarin 7-hydroxylation and the in-vivo Cl(po,u) for (S)-warfarin in relation to the CYP2C9*3 polymorphism. In conclusion, the in-vitro model for human CYP2C9*3 polymorphism using recombinant cytochrome P450 proteins would serve as a useful means for predicting changes in in-vivo kinetics for (S)-warfarin and possibly other CYP2C9 substrates in relation to CYP2C9*3 polymorphism.

Adolescent↗

Relationship between left ventricular hypertrophy and renal and retinal damage in untreated patients with essential hypertension.

We examined the relationship between left ventricular hypertrophy (LVH) and renal and retinal damage in 174 untreated patients with essential hypertension. As an index of renal and retinal damage, we examined proteinuria and retinal vascular change. LVH was diagnosed according to left ventricular mass obtained from echocardiography. Of the hypertensive patients, 111 patients (64%) had LVH. The incidences of proteinuria and advanced retinal vascular change were higher in patients with LVH than in those without LVH. In a multiple regression model, there was a significant positive correlation between left ventricular mass and proteinuria, as well as diastolic blood pressure, sex, age and body mass index. In conclusion, proteinuria is related to elevated left ventricular mass in patients with essential hypertension.

Blood Pressure↗

[A successful case of minimally invasive direct coronary artery bypass (MIDCAB) followed by PTCA].

A 70-year-old man who developed angina pectoris underwent cardiac catheterization, which showed total occlusion of the left anterior descending coronary artery (LAD), associated with 75% stenosis of the right coronary artery (RCA) and 90% stenosis of the small circumflex coronary artery (CX). The LAD received good collateral flow from the RCA. The patient was scheduled to undergo the MIDCAB for the LAD using the internal thoracic artery (ITA), combined with percutaneous transluminal coronary angioplasty (PTCA) for the RCA subsequently. A left anterior submammarian skin incision of 10 cm in length was made. The fifth costal cartilage was removed. The left ITA was directly harvested from the chest wall from the 4th to 7th intercostal space, and was anastomosed to the midportion of the LAD without cardiopulmonary bypass. The patient was quickly recovered after the operation. On the 8th postoperative day, the patient successfully underwent the PTCA for the RCA after the ITA-LAD graft had been verified to be patent. The MIDCAB could be indicated for multivessel coronary disease in conjunction with the PTCA.

Aged↗

Distribution of RGS4 mRNA in mouse brain shown by in situ hybridization.

RGS proteins (regulators of G protein signaling) are a newly identified group of proteins that negatively regulate G proteins. Some, including RGS4, have been shown to act as GTPase-activating proteins for several members of the Gi alpha subunits and the Gq alpha subunit. However, the signaling pathways of G protein-coupled receptors regulated by RGS4 are not well understood. We isolated the mouse RGS4 cDNA to investigate the roles of RGS4 protein in mouse brain. Expression of mouse RGS4 in yeast stimulated adaptation to mating pheromone, suggesting that it negatively regulates that G protein-mediated signaling pathway. Northern blot analysis of various mouse tissues revealed that RGS4 is expressed at high levels in brain, moderately low levels in heart, and very low levels in lung, liver, and skeletal muscle. In situ hybridization of mouse brain showed RGS4 mRNA mainly in the cerebral cortex, hippocampus, anterior olfactory nucleus, piriform cortex, olfactory tubercle, caudate-putamen, nucleus accumbens, islands of Calleja, substantia nigra, amygdala, the granular layer of cerebellum, middle cerebellar peduncle, and perifacial zone. These results suggest that RGS4 is expressed in various types of cells in the brain and may regulate multiple G protein-mediated signaling pathways.

Animals↗

Influence of donor age on the cytotoxicity and mutagenicity of ethylnitrosourea in cultured human T-lymphocytes.

The effects of age on the cytotoxicity and mutagenicity of N-ethyl-N-nitrosourea (ENU) in human peripheral blood T-lymphocytes were investigated using colony-forming assay in vitro. ENU was shown to induce a dose-dependent increase in cell killing and in mutation frequencies (MF). No significant correlation between age and ENU-induced 6-thioguanine-resistant (TGr) MF at the hypoxanthine phosphoribosyl transferase (HPRT) locus of the X-chromosome was found after treatment with the same concentration of ENU (1 mM or 2 mM). There were also no significant differences among different donor age groups and the sensitivity parameters for exposure to ENU. As X-rays, the cytotoxic and mutagenic effects of ENU in cultured human T-lymphocytes appear not to be associated with age. These results suggest that the repair of mutagen-induced DNA lesions does not decline with age. Such knowledge has implications for risk assessment and protection against environmental mutagens.

Adolescent↗

CMO I deficiency caused by a point mutation in exon 8 of the human CYP11B2 gene encoding steroid 18-hydroxylase (P450C18).

Corticosterone methyloxidase I (CMO I) deficiency is an autosomal recessive disorder of aldosterone biosynthesis. To determine further the molecular genetic basis of CMO I deficiency, a patient of Turkish origin that suffered from CMO I deficiency was studied. Nucleotide sequencing of the PCR-amplified exons from the genomic DNA of this patient revealed a single point mutation CTG (leucine) CCG (proline) at codon 461 in exon 8 of CYP11B2, which is involved in the putative heme binding site of steroid 18-hydroxylase (P450(C18)). The expression study using a cDNA introducing the point mutation revealed that the amino acid substitution totally abolishes the P450(C18)p3 enzyme activities required for conversion of 11-deoxycorticosterone to aldosterone, even though the mutant product was detected in the mitochondrial fraction of the transfected cells. These results suggest that this point mutation causes CMO I deficiency.

Aldosterone↗

Pulmonary hypertension caused by medial hypertrophy associated with aortic stenosis and preductal coarctation.

A 7-month-old female infant with aortic stenosis, preductal coarctation, and pulmonary hypertension underwent operation. Intraoperative lung biopsy revealed marked medial hypertrophy of the pulmonary arterioles. This histopathology is compatible with persistent pulmonary hypertension in the newborn. She is alive about 5 years after the operation, but pulmonary hypertension remains. The pathogenesis is discussed.

Aortic Coarctation↗

Alterations of insulin and glucagon secretion from the perfused pancreas before, at the onset and after the development of diabetes in male Otsuka Long-Evans Tokushima Fatty (OLETF) rats.

The Otsuka Long-Evans Tokushima Fatty (OLETF) rat is an obese non-insulin dependent diabetes mellitus (NIDDM) model of an inbred strain. In this study, basal (2.8 mM glucose) insulin and glucagon and their responses to glucose (16.7 mM) were examined at the age of 9 weeks (n = 3) before the onset of diabetes, at 23 weeks (n = 6) at the onset of diabetes, and at 48 weeks (n = 5) after the development of diabetes by pancreatic perfusion. In Long-Evans Tokushima Otsuka (LETO, control) rats, insulin responses to glucose showed a biphasic pattern at all three ages, while in OLETF rats, basal insulin concentrations were significantly increased compared to those in controls at the age of 9 and 48 weeks. Insulin responses to glucose showed no difference from controls at 9 and 23 weeks, however, at 48 weeks the response was significantly decreased. In controls, high basal glucagon concentrations showed significant decrease in response to glucose at all ages. In OLETF rats, basal glucagon concentrations showed significant decrease compared to those in control rats at 23 and 48 weeks. Glucagon response to glucose significantly decreased at 9 and 23 weeks, but at 48 weeks there was no change in concentration in response to glucose. Pancreatic insulin content was lower at 48 weeks in OLETF rats than in LETO rats, although no differences were observed at other ages. There were no significant differences in pancreatic glucagon content between the two groups at any age. Morphologically, in OLETF rats the number of pancreatic B cells were decreased and A cells migrated into the center of islets at 48 weeks. The results suggested that one of the causes of diabetes in OLETF rats is impaired insulin response to glucose.

Animals↗

Lymph node metastasis in carcinoma of the body and tail of the pancreas.

BACKGROUND: There have been no precise reports concerning lymph node metastatic involvement in carcinoma of the body and tail of the pancreas. METHODS: Histopathological examination of lymph node involvement in 30 specimens obtained from patients who underwent pancreatic resection and wide dissection of lymph nodes, including para-aortic lymph nodes, for carcinoma of the body and tail of the pancreas was performed. RESULTS: Fourteen of 30 patients had lymph node involvement. The highest incidence of lymph node involvement was around the splenic artery (five of 30 patients), aorta (four of 30) and coeliac trunk (four of 30). No significant difference in survival rate between the lymph node-negative group and the lymph node-positive group was observed, but all patients in the positive group died within 2 years after surgery, and four patients with para-aortic lymph node involvement died from recurrence within 10 months after surgery. Survival rates were significantly worse in patients with histopathological extrapancreatic nerve plexus invasion, retropancreatic tissue invasion, tumour diameter more than 4 cm, histological portal system vein wall invasion and carcinoma invasion of the surgical margins. CONCLUSION: Although aggressive extended surgery including para-aortic node dissection has been performed, the postoperative survival rate is still low in patients with carcinoma of the body and tail of the pancreas. The high incidence of liver metastasis after surgery is a prime cause of the poor outcome, and effective therapy for postoperative liver recurrence requires evaluation.

Adult↗

Functional analyses of mammalian protein kinase C isozymes in budding yeast and mammalian fibroblasts.

BACKGROUND: The PKC1 gene of Saccharomyces cerevisiae encodes a homologue of mammalian protein kinase C (PKC) that is required for yeast cell growth. Pkc1 has been proposed to regulate a protein kinase cascade which includes the Bck1, Mkk1/Mkk2 and Mpk1 kinases. The functional relationship between Pkc1 and mammalian PKCs is unknown. Another signal transduction in Saccharomyces cerevisiae, the mating pheromone signalling pathway, is mediated by a heterotrimeric G protein, and causes cell cycle arrest in the G1 interval. It is not clear whether PKC is involved in this pathway. The effects of overexpression of PKCs in mammalian cells have been widely studied to analyse the function of PKCs in vivo. RESULTS: We isolated a human cDNA which encodes a protein kinase C type eta (PKC-eta) by complementation of pkc1 mutations in Saccharomyces cerevisiae. The human PKC-eta was able to complement the growth defect caused by the deletion of PKC1, whereas PKC-eta was unable to suppress the defect caused by deletion of BCK1. We also isolated human cDNAs that can suppress the adaptation defect of sst2. One of them encodes a protein kinase C type delta (PKC-delta). Expression of this gene in yeast stimulated an adaptation to the pheromone response. Human PKC-delta suppressed the adaptation defect of a pheromone receptor mutation lacking its C-terminal domain, but not that of a G protein beta-subunit mutation eliminating signal-induced phosphorylation, and not the lethality of the gpa1 null mutation. Moreover, overexpression of PKC-eta in NIH3T3 cells induced anchorage-independent growth. CONCLUSIONS: PKC-eta has a biological activity which is closely related to Pkc1, and PKC-eta activates the Pkc1-mediated pathway through an activation of the Bck1 kinase that is a homologue of MAP kinase kinase kinase. PKC-eta appears to play a critical role in growth control of yeast and mammalian cells. Suppression experiments with PKC-delta suggest that PKC-delta desensitizes the pathway by regulating an aspect of G protein function.

Adaptation, Biological↗

Yolk sac tumor of the ovary in a conjoined twin.

A 5-year-old conjoined twin girl had a yolk sac tumor of the right ovary. She was admitted to our hospital because of abdominal distension. Comprehensive clinical and radiological investigations revealed a tumor of the right ovary. She underwent right salpingo-oophorectomy. The diagnosis was yolk sac tumor (stage I A), and postoperative chemotherapy (vincristine, actinomycine-D, cyclophosphamide) according to the UK-CCSG was provided. The postoperative course was uneventful with no evidence of recurrence 3 years after surgery. Her twin sister had no tumor. This is the first reported of yolk sac tumor of the ovary in a conjoined twin.

Child, Preschool↗

cDNA sequence and expression of a gene encoding a pyruvate dehydrogenase kinase homolog of Drosophila melanogaster.

Pyruvate dehydrogenase is a catalyst for an irreversible step in the degradation of glucose and its activity is regulated by a highly specific protein kinase, pyruvate dehydrogenase kinase (PDK). PDK belongs to a family of mitochondrial protein kinases unique from other eukaryotic protein kinases. We cloned a cDNA encoding a putative PDK from Drosophila melanogaster (DmPDK). The deduced DmPDK consists of 413 amino acids and shares up to 57.8% homology with human and rat PDK isoenzymes. Developmental Northern blot analysis revealed two major transcripts of 2.1 kb and 2.7 kb. The 2.7-kb transcript was expressed throughout ontogeny, whereas the 2.1-kb transcript was specific to embryos and adult females. Whole-mount in situ hybridization revealed that PDK mRNA is ubiquitously distributed in the embryo. The DmPdk gene was cytologically mapped to the 45CD region on the right arm of the second chromosome.

Amino Acid Sequence↗

Peritoneal washing cytology combined with immunocytochemical staining and detecting mutant K-ras in pancreatic cancer: comparison of the sensitivity and availability of various methods.

Peritoneal metastases are the second most common site of involvement, following the liver, in pancreatic cancer. Thus, we performed peritoneal washing cytology at laparotomy to diagnose accurately the intraperitoneal spread of carcinoma cells to determine the appropriate therapy. Peritoneal washings were collected at laparotomy from 20 Japanese pancreatic carcinoma patients at Nagoya University Hospital between April 1993 and December 1994. From centrifuged deposits, we examined the cytology by three methods as follows. The first method was conventional cytology, including May-Grünwald and Giemsa, Papanicolaou, periodic acid-Schiff, and Alcian blue. The second method was immunocytochemical staining, using antibodies to carbohydrate antigen (CA19-9) and carcinoembryonic antigen. After extracting DNA from the remaining pellet, we studied the last method, detecting K-ras point mutation, by two-step polymerase chain reaction and restriction fragment length polymorphism analysis. In two cases, peritoneal metastases were macroscopically recognized, and the results of all three methods were positive. In the two other cases, where peritoneal dissemination was not macroscopically recognized, the judgments of conventional cytological study and detecting K-ras point mutation were negative. However, a few malignant cells were found by the immunocytochemical staining method. Judging from their clinical course, the positively stained cells were suggestive of malignancy. At present, the immunocytochemical staining method is the most sensitive of these three methods in peritoneal washing cytology. However, preserving DNA is suitable for repeated examination, and a modified method can be applied. If the sensitivity increases, the method of detecting K-ras has the potential to become the standard for peritoneal washing cytology in pancreatic cancer.

Adult↗