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Biomedical subjects

T Morisaki

Publications and source records attributed to T Morisaki.

At least 73 records · Page 4Linked to original sources

Expression of MEF2 genes during human cardiac development.

To better understand the regulatory mechanisms in gene expression of human cardiomyocytes, we studied the expression of MEF2 genes encoding transcription factors during the course of cardiac development. Expression of all four MEF2 transcripts (MEF2A, MEF2B, MEF2C, and MEF2D) were detected in all developmental stage of the human heart, while Mef2b transcripts were down-regulated in mouse heart development. Although none of the MEF2 genes, besides mouse Mef2b, exhibited any remarkable quantitative change in their transcripts, qualitative changes in MEF2 transcripts were found during the course of cardiac development. In particular, MEF2D transcripts showed prominent changes by alternative splicing in the perinatal period. MEF2D transcripts containing the 21-base exon (exon b) were predominantly expressed after birth. At the same time, transcripts of the alpha myosin heavy chain (alphaMHC) gene increased after birth, as the splicing pattern in transcripts of the cardiac troponin T (cTnT) gene changed to decrease the transcripts of cTnT1 after birth. These changes seemed to be correlated with the alternative splicing changes of MEF2 genes, especially MEF2D. The alternative splicing as well as transcriptional regulation in MEF2 genes might be important for regulating the alphaMHC gene and the maturation of cardiomyocytes.

Animals↗

Positive washing cytology in patients with pancreatic cancer indicates a contraindication of pancreatectomy.

BACKGROUND: Although peritoneal washing cytology has been widely used in the field of gynecology, it has not been performed so frequently in patients with pancreatic cancer. Only a few papers have reported surgical implications of peritoneal washing cytology. MATERIALS AND METHODS: We reviewed results of peritoneal washing cytology performed immediately after laparotomy in 50 Japanese patients with pancreatic cancer. The 50 patients were divided into two groups according to the results of cytology. Clinicopathological findings were compared between these two groups. RESULTS: Cytology of peritoneal washing was positive in 13 (26%) of the 50 patients. Nine of the 13 patients had no visible peritoneal dissemination. There were no significant differences regarding the age, tumor size and serum level of the carcinoembryonic antigen. Moderately to poorly differentiated adenocarcinoma and perineural invasion (ne3) and lymphatic permeation (ly3) of marked degree were more frequent in the positive group than in the negative group (p < 0.01). The survival curve of the 37 patients with negative cytology was significantly better than that of the 13 with positive cytology (p < 0.01). Four of 13 patients with positive cytology underwent a potentially curative resection but died within 12 months. Three of these four patients died of peritonitis carcinomatosa with massive malignant ascites and one died of local recurrence. CONCLUSIONS: Positive peritoneal washing cytology is not rare in patients with pancreatic cancer and indicates a contraindication of pancreatectomy.

Adenocarcinoma↗

Novel biological functions of interleukin-4: formation of tube-like structures by vascular endothelial cells in vitro and angiogenesis in vivo.

The effect of interleukin-4 (IL-4) on angiogenesis was studied in vitro and in vivo. Human recombinant IL-4 significantly stimulated the formation of tube-like structures in collagen gels by bovine aortic endothelial cells as well as by human microvascular endothelial cells. Human recombinant IL-4 at 50-500 U/ml stimulated by about two- to threefold the formation of tubes by bovine aortic endothelial cells; the rate was comparable to that of basic fibroblast growth factor. Tube formation was almost completely inhibited by the addition of IL-4 receptor neutralizing antibody. Administration of rat recombinant IL-4 led to neovascularization when implanted in the rat cornea. Findings suggest that IL-4 may be a mediator of the immune system as well as an inducer of angiogenesis.

Animals↗

A novel gene, ITM, located between p57KIP2 and IPL, is imprinted in mice.

We searched for new imprinted genes using a positional cloning method in a region of human chromosome 11p15.5, which contains several imprinted genes including p57KIP2 and IPL, and found a novel ITM gene located between p57KIP2 and IPL. We also obtained the mouse homologue Itm in its syntenic region of mouse chromosome 7. In humans, its location is 17 kb centromeric to p57KIP2 and 3 kb telomeric to IPL, and in mice, 15 kb and 2.5 kb, respectively. They are expressed in most tissue, but especially in the kidney and liver, and moderately in the heart, lung and testis. Mice exhibit a functional imprinting resulting in higher expression of maternal alleles in fetal, newborn and most adult tissues, but it is biallelically expressed in the adult kidney and liver where expression is the highest. In addition to the discrepancy between the level of expression and the strength of the imprint, Itm has several unusual features for an imprinted gene, including large introns, moderate GC content and the absence of direct repeats. Our results will be helpful in understanding the intricate regulatory mechanism of imprinted genes.

Amino Acid Sequence↗

Identification of a novel human homolog of the Drosophila dlg, P-dlg, specifically expressed in the gland tissues and interacting with p55.

We have identified a novel human homolog of the Drosophila dlg tumor suppressor gene, termed P-dlg, which has been mapped at chromosome 10q23. Unlike other human dlg homologs, P-dlg is expressed in placenta and various gland tissues but not in brain. The P-dlg protein is localized at the plasma membrane and cytoplasm, and it is expressed in the gland epithelial cells in normal prostate tissue but not in prostate cancer cell lines. Furthermore, we identified interaction between P-dlg and p55 palmitoylated membrane protein by yeast two-hybrid screening. These findings suggest that P-dlg forms a complex with p55 at the plasma membrane and plays roles in maintaining the structure of epithelial cells and transmitting extracellular signals to the membrane and cytoskeleton, which may negatively regulate cell proliferation.

Amino Acid Sequence↗

Molecular detection of cancer cells by competitive reverse transcription-polymerase chain reaction analysis of specific CD44 variant RNAs.

BACKGROUND: CD44 is a cell surface glycoprotein implicated in such diverse biologic processes as lymphocyte activation and homing, extracellular matrix adhesion, and cellular migration. Primary transcripts of the CD44 gene can be alternatively spliced to produce a variety of messenger RNA (mRNA) species. The standard form of CD44 mRNA contains sequences from at least 20 genomic exons; variant mRNAs contain sequences from one or more additional exons (v1-10). Predominant expression of a specific CD44 variant, i.e., CD44v8-10, in several human carcinomas has been described previously. In this study, we developed a novel molecular approach for detecting cancer cells that overexpress CD44v8-10 mRNA. METHODS: After finding that CD44v8-10 was predominantly expressed in non-small-cell lung and bladder carcinomas and that CD44v10 was predominantly expressed in leukocytes, we developed a competitive reverse transcription-polymerase chain reaction assay (CC-RT-PCR) that allows quantification of the relative expression of these two mRNA species in clinical specimens (i.e., determination of a v8-10/v10 ratio). CC-RT-PCR analysis was applied to pleural effusion specimens from patients with benign or malignant lung diseases as well as to spontaneously voided urine samples from patients with benign or malignant urologic diseases. RESULTS: Fifty two of 54 samples from patients with benign diseases expressed CD44v10 predominantly (v8-10/v10 ratio < or = 0.65), whereas 46 of 61 samples from patients with malignant diseases expressed CD44v8-10 predominantly (v8-10/v10 ratio > 1.00) (two-sided P < .001). CC-RT-PCR detected predominant expression of CD44v8-10 in cytologically negative samples from 11 patients who were later diagnosed with malignant disease. CONCLUSIONS: CC-RT-PCR analysis of CD44v8-10 expression could be an important adjunct to cytologic examination in cancer diagnosis, especially in detecting exfoliated cancer cells in pleural effusions and urine.

Breast Neoplasms↗

N-glycosylation does not affect assembly and targeting of proglycinin in yeast.

Glycinin, a simple protein, and beta-conglycinin, a glycoprotein, are the dominant storage proteins of soybean and are suggested to be derived from a common ancestor. To investigate why glycinin does not require glycosylation for its maturation, we attempted N-glycosylation of proglycinin A1aB1b using site-directed mutagenesis and yeast expression system. An N-glycosylation consensus sequence Asn-X-Ser/Thr was created at positions 103, 183, 196, 284 and 457 in the variable regions being strongly hydrophilic revealed from the alignment of amino acid sequences of various glycinin-type proteins. Among five mutant proglycinins (Q103N, H183N, G198T, S284N, N459T), Q103N was fully glycosylated, and H183N and N459T were partly (around 20% of the expressed proteins), whereas others were barely or not glycosylated. The glycosylated proglycinin was susceptible to endo-beta-N-acetylglucosamidase and N-glycanase cleavages. N-glycosylation did not cause inconveniences to processing of signal peptide, assembly into trimers and targeting into the vacuoles. Thermal and trypsin sensitivity analyses of the glycosylated proglycinin suggested that N-linked glycan prevents protein-protein interaction but does not stabilize the protein conformation. The reason why glycinin does not require N-glycosylation for its maturation is discussed.

Antigens, Plant↗

Coil embolization of bleeding visceral pseudoaneurysms following pancreatectomy: the importance of early angiography.

OBJECTIVE: To evaluate the efficacy of early angiography in the diagnosis and treatment of delayed arterial hemorrhage as a significant complication following pancreatectomy. DESIGN: Retrospective case series. SETTING: University hospital; 6-year period between January 1992 and December 1997. PATIENTS AND INTERVENTION: Of a total of 81 patients undergoing pancreatic head resection, 10 (12%) developed massive arterial hemorrhage after surgery. All 10 patients underwent an emergency angiography and 8 of these were managed by transcatheter arterial embolization. RESULTS: Before onset of major bleeding, 9 patients (90%) had developed intra-abdominal abscess resulting from pancreatic fistula or other anastomotic leak, and all 10 patients had had preliminary minor bleeding. The angiogram demonstrated an exact site of bleeding as a pseudoaneurysm in all 10 patients. Transcatheter arterial embolization achieved temporary control of bleeding in all 8 patients for whom embolization was attempted and complete hemostasis in 5 of the 8 subsequently, yielding a success rate of 63%. Overall, 4 patients (40%) died of complications related directly to the major hemorrhage or the transcatheter arterial embolization technique. CONCLUSIONS: An emergency angiography should be considered in all patients who develop either a sentinel or massive gastrointestinal bleed following pancreatectomy irrespective of suspected intra-abdominal sepsis. Transcatheter arterial embolization allows temporary control of massive hemorrhage and hemodynamic stabilization in most cases, and prevents the need for high-risk emergency surgery.

Aged↗

Expression of interleukin (IL)-12 mRNA in gastric carcinoma specimens: cellular antitumor immune responses.

BACKGROUND AND OBJECTIVES: Several tumor-related antigen peptides that are recognized by autologous cytolytic T cells (CTL) have been reported. However, most human solid tumors, including gastric carcinoma, are only weakly immunogenic. In this study, we focused on interleukin (IL)-12 and interferon-gamma (IFN-gamma) as key cytokines for estimating positive cellular immune responses. METHODS: To estimate the immunogenicity of gastric carcinomas, we examined IL-12 and IFN-gamma at mRNA levels by reverse transcription-polymerase chain reaction assay (RT-PCR) in tumor specimens and adjacent nontumor specimens from 36 gastric carcinoma patients. RESULTS: IL-12 expression was detected in 12 tumor specimens and in only two adjacent nontumor specimens (P = .003). The frequency of IFN-gamma gene expression was higher in the IL-12 mRNA-positive tumor specimens than in the IL-12 mRNA-negative tumor specimens (P = .015). In the IL-12 mRNA-positive tumors, IFN-gamma expression was higher in the tumor specimens than in the adjacent nontumor specimens (P = .007). Conversely, in the IL-12 mRNA-negative tumors, IFN-gamma expression was lower in the tumor specimens than in the nontumor specimens (P = .03). Many tumor-infiltrating mononuclear cells, predominantly T cells, were found in four of the 12 IL-12-mRNA-positive tumor specimens and in none of the 24 IL-12-mRNA-negative tumor specimens (P = .008). CONCLUSIONS: These data suggest that possible immune responses against a tumor may occur at the mRNA level in approximately one-third of gastric carcinomas.

Blotting, Southern↗

Expression of inducible nitric oxide synthase in circulating neutrophils of the systemic inflammatory response syndrome and septic patients.

There is increasing evidence that nitric oxide (NO) is an important factor in the pathogenesis of septic shock. It is known that polymorphonuclear neutrophils (PMNs) are activated during sepsis or after surgical stress, and they then release various toxic mediators including free radicals. It has not been clear whether NO synthesis can be induced in circulating PMNs. Blood samples were obtained from 11 patients with sepsis, 23 patients with systemic inflammatory response syndrome (SIRS), and 16 patients without SIRS (nonSIRS) who underwent operation. We examined mRNA expression of inducible NO synthase (iNOS) in circulating PMNs from those patients pre- and postoperatively using the reverse transcriptase polymerase chain reaction (RT-PCR) method and measured their serum nitrate (NO2-) + nitrate (NO3-) concentration, peripheral blood white cell (WBC) count, and serum C-reactive protein (CRP) level. The frequency of iNOS expression in PMNs increased in sepsis (100%) and SIRS (70%) patients compared to that in nonSIRS patients (18%) (p < 0.001). The peripheral WBC count and CRP level were significantly higher in iNOS-positive patients than in iNOS-negative patients (p < 0.05 and p < 0.01, respectively). Postoperatively, the serum NO2- + NO3- concentration increased in 87% of septic patients and in 56% of patients with SIRS (p < 0.05 for both). Our study indicated that iNOS mRNA expression is induced in human circulating PMNs of patients with postoperative sepsis and SIRS and may be involved in the pathogenesis of the sepsis syndrome.

Adult↗

Comparison of endoscopic retrograde and magnetic resonance cholangiopancreatography in the surgical diagnosis of pancreatic diseases.

BACKGROUND: Magnetic resonance cholangiopancreatography (MRCP) is a newly developing noninvasive examination of the biliopancreatic trees. Roles of MRCP in the diagnosis of pancreatic diseases have not been scrutinized. METHODS: Endoscopic retrograde cholangiopancreatography (ERCP) and MRCP were reviewed in 52 Japanese patients with various pancreatic diseases and 6 patients with normal pancreas to compare their diagnostic usefulness and limitation. RESULTS: In those with normal pancreas, only the main pancreatic duct was visualized by MRCP, while both the main pancreatic and branch ducts were clearly delineated by ERCP. In 3 patients with serous cystadenoma, the tumor was not visualized by ERCP, whereas it was visible as a high-intensity mass on MRCP. Of 18 patients with a "mucin hypersecreting" tumor of the branch type, MRCP demonstrated cystically dilated branch ducts in all, while ERCP failed to visualize the dilated ducts in 6 patients. However, the details of the cystic lesions (mural nodule, communication with the main pancreatic duct) were more exactly demonstrated by ERCP than MRCP. In 5 patients with a mucin hypersecreting tumor of the main pancreatic duct type, the dilated main pancreatic duct and the presence of mural nodules were similarly demonstrated both by ERCP and MRCP. In 12 patients with pancreatic adenocarcinoma, indirect findings were similarly demonstrated both by ERCP and MRCP, ie, stenosis (4 patients) and obstruction (8) together with dilation of the main pancreatic duct (9). In 3 patients, the center of the mass showed high intensity on MRCP, suggesting the secondary change of pancreatic carcinoma. In 8 patients with obstruction of the main pancreatic duct due to carcinoma, the distal pancreatic duct was visualized by MRCP but not by ERCP. In 9 patients who had undergone pylorus-preserving or standard pancreatoduodenectomy, follow-up MRCP was obtainable in all examined and displayed the main pancreatic duct. CONCLUSIONS: MRCP plays a complementary role in the surgical diagnosis of pancreatic disorders and is especially useful to examine the pancreatic duct after pancreatoduodenectomy.

Adenocarcinoma↗

Assessment of c-erbB2 and vascular endothelial growth factor mRNA expression in fine-needle aspirates from early breast carcinomas: pre-operative determination of malignant potential.

AIMS: Although axillary lymph nodes status, tumour size, hormonal-receptor status and histological grade at diagnosis are frequently used to orient the treatment of breast cancer patients, some tumours recur in patients with early stage disease. Pre-operative assessment of individual tumour characteristics, based on oncogenes and growth factors related to tumour growth, invasion or metastasis, may guide the treatment for patients with breast carcinomas. METHODS: We examine here the prognostic significance of cyclin D1, urokinase type plasminogen activator, vascular endothelial growth factor (VEGF), platelet-derived growth factor, and c-erbB2 expression in pre-operatively obtained fine-needle aspirates from breast carcinomas less than or equal to 3 cm in size. Correlation between mRNA expression of these factors and clinicopathological characteristics was analysed. RESULTS: The level of c-erbB2 mRNA expression was significantly higher in tumours with lymph node metastases than in those without lymph node metastases. VEGF mRNA expression positively correlated with the degree of angiogenesis as quantitated by immunohistological staining with a CD31 monoclonal antibody. CONCLUSIONS: Analysis of c-erbB2 and VEGF mRNA expression in fine-needle aspirates may be useful in assessing the malignant potential of individual breast carcinomas, leading to a pre-operative discrimination of a high-risk group.

Actins↗

Short-term and long-term pancreatic exocrine and endocrine functions after pancreatectomy.

Exocrine and endocrine functions of the pancreas were assessed in 44 Japanese patients who underwent pancreatic head resection. Functions were analyzed comparing levels before surgery, at a short-term follow-up (<2 months), and at a long-term follow-up (12-31 months). The N-benzoyl-L-tyrosyl-p-aminobenzoic acid (BT-PABA) excretion test, fasting blood sugar (FBS) level, and oral glucose tolerance test (OGTT) were used to determine pancreatic function. The patients were divided into three groups according to the size of the main pancreatic duct: group 1, 15 patients with a normal sized duct (< or =3 mm); group 2, 20 with a moderately dilated duct (>3 mm, <10 mm); and group 3, 9 with a markedly dilated duct (> or =10 mm). The mean BT-PABA value (6-hr urinary PABA recovery rate) in group 1 showed no change during the postoperative period. In contrast, the BT-PABA values in groups 2 and 3 had dropped by the short-term follow-up and returned to the preoperative level by the long-term examination. FBS and 120-min OGTT levels were not different between the three groups preoperatively. Although these values showed no change in all the three groups at the short-term measurements, the FBS in group 3 and 120-min levels in all the three groups had increased at the long-term. These findings suggest that exocrine pancreatic function shows a short-term deterioration in patients with a dilated pancreatic duct but recovers to the preoperative level over the long term after pancreatic head resection. Endocrine insufficiency, however, may occur at a long-term point after surgery irrespective of the preoperative pancreatic ductal dilatation.

4-Aminobenzoic Acid↗

Transforming growth factor beta1 (TGF-beta1) is a preoperative prognostic indicator in advanced gastric carcinoma.

It has been generally accepted that transforming growth factor beta1 (TGF-beta1) has both negative and positive effects on tumour growth and progression. This study analysed the prognostic value of TGF-beta1 mRNA in advanced gastric carcinoma. A reverse transcriptase-polymerase chain reaction analysis (RT-PCR) was used for TGF-beta1 in endoscopic biopsy specimens from 42 advanced gastric carcinomas. Thirty specimens expressed TGF-beta1 mRNA while 12 specimens did not. The follow-up duration ranged from 4 to 37 months (mean 22.8 months). TGF-beta1-positive group demonstrated a shorter overall survival compared with the TGF-beta1 -negative group (P=0.0014). A significant correlation was also found in the 32 patients who underwent curative resection (P=0.0048). Significant correlations were found between TGF-beta1 mRNA expression and both stage (P=0.0015) and nodal involvement (P=0.0060). Multivariate analysis demonstrated that only TGF-beta1 mRNA expression (P=0.0306) was an independent prognostic factor. All of ten patients who underwent non-curative resection expressed TGF-beta1 mRNA. Expression of TGF-beta1 mRNA in gastric biopsy specimens may be an important preoperative prognostic variable for advanced gastric carcinoma.

Aged↗

Prognostic value of platelet-derived growth factor-A (PDGF-A) in gastric carcinoma.

OBJECTIVE: Because our previous study indicated that PDGF-A mRNA expression in biopsy specimens might identify a subgroup of high-risk patients with gastric carcinoma, in this study we analyzed the prognostic value of platelet-derived growth factor-A (PDGF-A) gene expression in gastric carcinoma biopsy specimens. METHODS: Reverse transcriptase-polymerase chain reaction (RT-PCR) was used to analyze the PDGF-A gene expression in 65 gastric carcinoma endoscopic biopsy specimens. The 65 patients were divided into a PDGF-A-positive group (29 patients) and a PDGF-A-negative group (36 patients). RESULTS: On the basis of 2-year follow-up data, the PDGF-A-positive group demonstrated a shorter overall survival rate compared with the PDGF-A-negative group (p < 0.0001). A similar correlation was found in 34 advanced-stage patients (p = 0.003) and in 24 advanced-stage patients who underwent a curative resection (p = 0.003). Multivariance analysis indicated that the transcription of PDGF-A gene is a potent prognostic factor that is independent of the traditional pathologic parameters. CONCLUSIONS: Expression of PDGF-A mRNA in gastric biopsy specimens may be a new preoperative prognostic parameter in gastric carcinoma.

Aged↗

Pleomorphic carcinoma of the pancreas: reappraisal of surgical resection.

Pleomorphic carcinoma is a rare variant of pancreatic exocrine carcinoma. The aim of this communication is to reappraise surgical resection of pleomorphic carcinoma of the pancreas. Clinicopathological findings of four Japanese patients with pleomorphic carcinoma of the pancreas were reviewed and compared with those of 24 Japanese patients with adenocarcinoma of the pancreas to clarify possible surgical implications of pleomorphic carcinoma. Of the four patients, three were female and one male, aged 64, 65, 66, and 74 yr, respectively. Two carcinomas were located in the head of the pancreas, one in the body, and the other in the tail. Ultrasonography demonstrated a well defined hypoechoic mass measuring 5-10 cm, with central necrotic area in all of the patients. Computed tomography showed a low density tumor with sharp margin and heterogeneous internal structure in all. On angiography, three tumors were hypervascular and another was hypovascular. Extensive vascular encasement was observed in all. Pancreatoduodenectomy was done in two patients and distal pancreatectomy in the other two. Multiple liver metastases occurred 1 month after surgical resection in two patients and local recurrence 1 month in one and 2 months in the other, leading to death either 2 (2 patients) or 3 months (2 patients) after pancreatectomy. Significantly differentiating features of the four pleomorphic carcinomas of the pancreas and the 24 adenocarcinomas of the pancreas were the mean diameter (6.6 +/- 1.3 cm vs 3.5 +/- 0.3 cm, p = 0.0007), margin of the tumor (expansive in the four pleomorphic carcinomas versus infiltrative in 21 of the 24 adenocarcinomas, p = 0.003) and vascularity on angiography (hypervascular in three of the four pleomorphic carcinomas versus hypovascular in 21 of the 23 adenocarcinomas, p = 0.013). The 1-yr and 3-yr survival rates of the four patients with pleomorphic carcinoma were 0% and 0%, whereas those of the 24 patients with adenocarcinoma of the pancreas were 42% and 13%, respectively (p < 0.0001). These findings suggest that the clinical course of patients with pleomorphic carcinoma of the pancreas is so poor even after surgical resection that pleomorphic carcinoma of the pancreas is not a candidate for pancreatectomy despite its locally expansive growth.

Adenocarcinoma↗

Control of AMP deaminase 1 binding to myosin heavy chain.

AMP deaminase (AMPD) plays a central role in preserving the adenylate energy charge in myocytes following exercise and in producing intermediates for the citric acid cycle in muscle. Prior studies have demonstrated that AMPD1 binds to myosin heavy chain (MHC) in vitro; binding to the myofibril varies with the state of muscle contraction in vivo, and binding of AMPD1 to MHC is required for activation of this enzyme in myocytes. The present study has identified three domains in AMPD1 that influence binding of this enzyme to MHC using a cotransfection model that permits assessment of mutations introduced into the AMPD1 peptide. One domain that encompasses residues 178-333 of this 727-amino acid peptide is essential for binding of AMPD1 to MHC. This region of AMPD1 shares sequence similarity with several regions of titin, another MHC binding protein. Two additional domains regulate binding of this peptide to MHC in response to intracellular and extracellular signals. A nucleotide binding site, which is located at residues 660-674, controls binding of AMPD1 to MHC in response to changes in intracellular ATP concentration. Deletion analyses demonstrate that the amino-terminal 65 residues of AMPD1 play a critical role in modulating the sensitivity to ATP-induced inhibition of MHC binding. Alternative splicing of the AMPD1 gene product, which alters the sequence of residues 8-12, produces two AMPD1 isoforms that exhibit different MHC binding properties in the presence of ATP. These findings are discussed in the context of the various roles proposed for AMPD in energy production in the myocyte.

AMP Deaminase↗