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Biomedical subjects

H Goto

Publications and source records attributed to H Goto.

At least 307 records · Page 17Linked to original sources

A RING finger motif regulates transforming activity of the rfp/ret fusion gene.

The rfp/ret transforming gene was generated by DNA rearrangement which occurred during transfection of NIH 3T3 cells. The amino-terminal half of Rfp with a RING finger motif was fused to the truncated Ret receptor tyrosine kinase, leading to a transforming sequence, Rfp/Ret. In the present study, to elucidate the importance of a RING finger motif for its transforming activity, we mutated cysteine or histidine residues present in the motif that are believed to be crucial for the tertiary structure of the protein. Substitution of Cys-16, Cys-31, or His-33 markedly decreased the transforming activity of Rfp/Ret. These mutations also resulted in a striking decrease of tyrosine phosphorylation of the mutant proteins in transfectants. In contrast, mutations of Cys-118 or His-124 in the second cysteine/histidine-rich motif (called B box) of Rfp/Ret did not affect its activity. These results thus indicated that the RING finger structure is critical for the full transforming activity of Rfp/Ret.

3T3 Cells↗

Intraabdominal contamination after gallbladder perforation during laparoscopic cholecystectomy and its complications.

BACKGROUND: Gallbladder perforation often occurs during laparoscopic cholecystectomy. METHODS: The frequency and causes of gallbladder perforation as well as the relevant clinical background factors were investigated in 110 patients undergoing laparoscopic cholecystectomy. We also evaluated intraperitoneal contamination by bacteria and gallstones at the time of gallbladder perforation and investigated whether perforation caused early or late postoperative complications. RESULTS: Intraoperative gallbladder perforation occurred in 29 of the 110 patients (26.3%). It was caused by injury with an electric knife during dissection of the gallbladder bed, injury during gallbladder retraction with grasping forceps, injury during gallbladder extraction from the abdomen, and slippage of cystic duct clips (potentially causing bile and stone spillage). Perforation was more frequent in patients with positive bile cultures and in those with pigment stones (p < 0.02), but not in patients with cholecystitis or cystic duct obstruction. The peritoneal cavity was contaminated by bacteria in 11/29 patients (37.9%) and by spilled stones in 3/29 patients (10.3%). There was no difference in the incidence of postoperative complications between the patients with and without perforation either in the early postoperative period or during follow-up for 24-42 months. Only one patient developed abdominal pain and fever in the early postoperative period, and they were probably related to perforation. CONCLUSIONS: Although gallbladder perforation is sometimes unavoidable during laparoscopic cholecystectomy, the risk of severe complications appears to be minimized by early closure of perforation, retrieval of as many of the spilled stones as possible, and intraperitoneal lavage.

Cholecystectomy, Laparoscopic↗

Inflammatory reactions after vascular prosthesis implantation: a comparison of gelatin-sealed and unsealed Dacron prostheses.

Despite widespread use of the gelatin-sealed knitted Dacron prosthesis (GDP) in clinical practice owing to its zero porosity, the biological impacts of this graft are still controversial. We conducted a randomized controlled study on 50 patients undergoing abdominal aortic aneurysm repair to evaluate the inflammatory reaction to GDP (n = 25) and unsealed knitted Dacron prostheses (UDP, n = 25). There were no significant differences in the mean age, size of the aneurysm, operative time, blood loss, or transfusion requirements between the GDP and UDP groups. During the first 7 postoperative days (PODs), slight fever and leukocytosis were noticed in both groups. Significant differences in maximum body temperature, leukocyte count, and plasma C-reactive protein concentration were observed between the GDP and UDP groups on POD 14:37.2 +/- 0.5 degrees C vs 36.9 +/- 0.3 degrees C (P = 0.019), 8,151 +/- 1,788/microliters vs 6,914 +/- 1,501/microliters (P = 0.015), and 32.6 +/- 27.5 mg/l vs 19.0 +/- 15.8 mg/l (P = 0.048), respectively. By POD 21, however, there were no detectable differences in these variables. Thus, we concluded that GDP caused an inflammatory reaction in the 2nd week after implantation, but ultimately there were no significant differences from UDP by the 3rd week.

Aged↗

Idiopathic portal hypertension associated with high serum titer of autoantibodies and liver dysfunction.

Idiopathic portal hypertension (IPH) is a condition marked by unexplained portal hypertension. Although a number of immunological abnormalities occur in patients with IPH, liver function is usually normal. We experienced an unusual case of IPH in a 49-year-old woman, who had pronounced splenomegaly. Laboratory data revealed pancytopenia, hypergammaglobulinemia, and liver dysfunction. Antinuclear antibodies were positive, with high titer at 1280 dilutions of sera. LE cell phenomena were also positive. Histological examination of biopsied liver showed only mild changes, but portal venous pressure was markedly elevated, at 38 cm H2O. This case was thus characterized by both a high serum titer of autoantibodies and liver dysfunction.

Autoantibodies↗

Mapping of antigenic sites on the major inner capsid protein of avian rotavirus using an Escherichia coli expression system.

The cDNA encoding the VP6 gene of avian rotavirus PO-13 strain was inserted into the bacterial expression vector pET-3a. Upon isopropyl-1-thio-beta-D-galactoside induction, the E. coli BL21 (DE3) harboring the vector containing cDNA of the VP6 gene produced an approximately 45-kDa polypeptide, which reacted with rabbit serum against PO-13 strain in Western blotting. To study the antigenic sites on VP6, various deletion mutants were constructed, expressed in E. coli and the reactivity with antigenic site I- and II-specific MAbs analyzed by Western blotting. Site I, which is shared with all group A mammalian and avian rotaviruses except for chicken rotavirus, was found to be located at amino acid positions 45 to 65, and site II, which probably contributes to an authentic group A antigen common to both mammalian and avian rotaviruses, at amino acid positions 134 to 142.

Animals↗

A novel de novo mutation in HPRT gene responsible for Lesch-Nyhan syndrome (HPRT OSAKA).

A virtually complete deficiency of hypoxanthine guanine phosphoribosyltransferase (HPRT) causes Lesch-Nyhan syndrome. A novel mutation of HPRT gene in a Japanese Lesch-Nyhan family has been identified using mRNA and genomic DNA from peripheral blood cells. A single nucleotide substitution of T to C in exon 3 resulted in a mis-sense mutation, CTC (Leu) to CCC (Pro), at codon 65. Utilizing an Mn/I restriction site which was lost in the mutation as an indicator, a family study showed that the mother was normal not having the mutant gene. The mutation was a de novo event that had occurred in the germ cells of the mother or in the proband during the early phase of fetal development.

DNA↗

t(7;11) and trilineage myelodysplasia in acute myelomonocytic leukemia.

A 48-year-old Japanese man was admitted to our hospital because of general fatigue, nasal bleeding, and petechiae on his extremities. He was diagnosed with acute myelomonocytic leukemia with trilineage myelodysplasia (T-MDS). Chromosomal analysis of bone marrow cells revealed t(7;11)(p15;p15), which has been rarely reported but known to be characteristic of Japanese patients. Although t(7;11)(p15;p15) has been reported mainly in acute myelogenous leukemia (AML), it can be occasionally found in so-called stem cell diseases such as chronic myelogenous leukemia or chronic myeloproliferative disorders. Therefore, t(7;11)(p15;p15) might affect trilineage progenitors or stem cells as well as myeloid lineage cells, subsequently resulting in AML with T-MDS, as in our case reported here.

Antineoplastic Combined Chemotherapy Protocols↗

The distribution of G and P types within isolates of bovine rotavirus in Japan.

Various combinations of G type and P type were observed in 76 bovine rotavirus (BRV) strains isolated from 235 diarrheal calves in three prefectures of Japan in 1992-1994. The most prevalent combination was G6:P5 (46/76, 60.5%), followed by G10:P11 (13/76, 17.1%), G6:P11 (7/76, 9.2%) and G10:P5 (5/76, 6.6%). No G6:P1 strain of BRV was recognized from the isolates in the present study, though this type of BRV is well known as a suitable vaccine strain against BRV infection.

Animals↗

Expression of the major inner capsid protein, VP6, of avian rotavirus in mammalian cells.

A gene encoding the major inner capsid protein, VP6, of avian rotavirus was inserted into the eukaryotic expression vector pAX-91 under the control of the SR alpha promoter and was expressed at a high level in simian COS7 cells. The expressed VP6 was indistinguishable in terms of electrophoretic mobility from the corresponding protein synthesized in simian MA104 cells infected with avian rotavirus. Binding assays with a series of monoclonal antibodies (mAbs) that corresponded to four antigenic sites on VP6 of avian rotavirus showed that the antigenic characteristics of the expressed product were identical to those of the native VP6 of avian rotavirus virions. Fiber-like structures that reacted strongly with antiserum against rotavirus were observed in VP6-expressing COS7 cells. Furthermore, an analysis of the tertiary structure of the expressed VP6 protein indicated that it adopts a trimeric configuration, similar to that of the major inner capsid protein of PO-13 virus. From these results, it appears that recombinant VP6 will facilitate studies of the structure and function of authentic VP6, an important protein in avian rotavirus.

Animals↗

Preoperative staging of superficial esophageal carcinoma: comparison of an ultrasound probe and standard endoscopic ultrasonography.

BACKGROUND: In diagnosing superficial esophageal carcinoma, it is necessary to differentiate mucosal carcinoma from submucosal carcinoma because mucosal carcinoma has a good prognosis and local treatment is likely to be successful. We evaluated an ultrasound probe and endoscopic ultrasonography (EUS) in the staging of superficial esophageal carcinoma. METHODS: From October 1992 to September 1994, 22 patients with 25 lesions (7 mucosal carcinomas, 18 submucosal carcinomas) were examined preoperatively with both the probe and EUS. The ultrasound findings were compared with histologic findings in all cases. RESULTS: The accuracy rates of the depth of invasion by the ultrasound probe were 86% (6 to 7) for mucosal carcinoma and 94% (17 to 18) for submucosal carcinoma, total 92% (23 to 25); by EUS 71% (5 to 7) for mucosal carcinoma and 78% (14 to 18) for submucosal carcinoma, total 76% (19 to 25). In the evaluation of lymph node metastasis, the overall accuracy was 56% by the ultrasound probe (sensitivity 25% and specificity 80%) and 67% by EUS (sensitivity 50% and specificity 80%). CONCLUSIONS: The ultrasound probe was more convenient to use and more accurate than EUS in the evaluation of the depth of invasion of superficial esophageal carcinoma.

Adult↗

Hepatocyte growth factor levels in bone marrow plasma of patients with leukaemia and its gene expression in leukaemic blast cells.

Hepatocyte growth factor (HGF) has been known as a multiple function factor, which also stimulates early haematopoiesis. In this study, we found that HGF was expressed at both the RNA and protein levels in acute myeloid leukaemia (AML) and chronic myeloid leukaemia (CML). In patients with AML (n = 20) and CML (n = 5), bone marrow plasma HGF concentrations were 20.44 +/- 6.26 (mean +/- s.e.) ng ml-1 and 7.17 +/- 0.53 ng ml-1 respectively. These were significantly higher (P < 0.01) than the value for normal subjects (n = 26): mean 0.92 +/- 0.09 ng ml-1. Constitutive HGF production was observed in freshly prepared leukaemic blast cells from three patients with high HGF levels of bone marrow plasma. Expression of HGF mRNA was correlated with bone marrow plasma HGF levels. After complete remission was obtained in six patients, bone marrow plasma HGF levels were significantly decreased. In contrast, the HGF mRNA was less abundantly expressed in acute lymphoid leukaemia (ALL). In patients with ALL (n = 5), bone marrow plasma HGF concentration (0.69 +/- 0.14 ng ml-1) remained low within the value for normal subjects. These results suggest that some populations of myeloid lineage cells have the ability to produce HGF.

Adult↗

Implication of nitric oxide synthase activity in the genesis of water immersion stress-induced gastric lesions in rats: the protective effects of FK506.

BACKGROUND: Nitric oxide (NO) is a potent cytoprotective substance of gastric mucosa. FK506, an immunosuppressive drug, shows anti-gastric ulcer effects equivalent to famotidine, an H2 blocker, in rats. This study was designed to evaluate the cytoprotective mechanism of FK506 on gastric mucosa in relation to the changes in NO synthase activity. METHODS: Gastric lesions were induced in rats by water immersion stress. Changes in NO synthase activity during water immersion stress treatment, and effects of FK506 on NO synthase activity were determined enzymatically. Gastric mucosal interleukin (IL)-1 beta and IL-2 were measured by immunoradiometric assay. Gastric mucosal blood flow was measured by hydrogen gas clearance technique. RESULTS: FK506 mitigated gastric lesions developed by water immersion stress. Stress-induced lesions were exacerbated by NG-monomethyl-L-arginine (L-NMMA), a specific inhibitor of NO synthase, while sodium nitroprusside, a NO donor, mitigated the lesions. Water immersion stress increased NO synthase activity in the early phase (0.5 h after stress treatment) and decreased it in the late phase (6 h after). Decrease in NO synthase activity in the late phase was significantly mitigated by FK506, though it did not affect changes in NO synthase activity in the early phase. Water immersion stress increased gastric mucosal IL-1 beta and IL-2 contents 6 h after stress treatment, and these increases were prevented by FK506. FK506 itself did not affect gastric mucosal blood flow. L-NMMA treatment significantly decreased gastric mucosal blood flow. In contrast, gastric mucosal blood flow was significantly increased by sodium nitroprusside. CONCLUSIONS: Increase in NO synthase activity might contribute to cytoprotection, and a decrease in activity might be a harmful factor for the gastric mucosa. Preservation of NO synthase activity by FK506 might be involved in FK506's protective effects on the gastric mucosa.

Animals↗

The role of complement in the acute inflammatory process in the skin and in host-parasite interaction in hamsters inoculated with Leishmania (Leishmania) chagasi.

Tecidual reaction at the inoculation site of L. (L.)chagasi promastigotes in hamsters depleted and non-depleted of complement was studied within 2, 6, 12, 24, 48 and 72 hours of infection. The inflammatory reaction was characterized by early predominance of polymorphonuclear cells (PMN) at 2, 6 and 12 hours of infection, mixed infiltrate of PMN and mononuclear cells (MN) at 24 hours, followed by predominance of MN at 48 and 72 hours after infection. The group depleted of complement showed a higher number of PMN at 2 hours and lower numbers of MN at 72 hours after infection (P < 0.0001). In the depleted group the phagocytosis by PMN was lower at 2 and 24 hours and by MN was lower at 24, 48 and 72 hours after infection. Electron microscopy showed extracellular intact and degenerated parasites, and lysed intracellular parasites, in PMN; and, rarely, preserved intracellular parasites in MN at 2, 6 and 12 hours after infection. The groups examined at 24, 48 and 72 hours of infection showed only cellular and parasite debris in mononuclear inflammatory cells. C3b deposits were detected by immunofluorescence in the interstitium and in the cytoplasm of inflammatory cells in non-depleted group at 2, 6 and 12 hours of infection. No immunoglobulin was detected in either group. Visceralization was detected 240 days after infection. The complement system has an important role in the inflammatory reaction and phagocytosis. The ultrastructural findings showed that the escape of the parasite probably occurs soon after inoculation.

Acute Disease↗

Soluble CD4: a link between specific immune mechanisms and non-specific inflammatory responses?

The CD4 molecule is an important cell surface molecule expressed on many cell types including T-helper cells. CD4 can be shed from cells, and the truncated soluble form of CD4 (sCD4) has been found elevated in serum in several diseases. In this study the authors show that migration of polymorphonuclear neutrophils (PMN) is induced by sCD4. Histopathology of the site of injection of recombinant sCD4 in the skin shows local infiltration of PMN. In vitro, in Boyden chamber, sCD4 can rapidly give rise to a dose-dependent migration of PMN. The authors speculate that sCD4 can be another chemotactic factor and, as such, constitutes a link within the immune system between specific and non-specific elements of inflammation.

Animals↗

Endothelium-dependent vasodilator effect of extract prepared from the roots of Paeonia lactiflora on isolated rat aorta.

Paeoniae Radix (the roots of Paeonia lactiflora Pallas) is a crude drug that is used in Asia and Europe to improve blood flow. We studied its vasodilator effect and mechanisms of action in vitro. The extract from Paeoniae Radix (PRE) relaxed prostaglandin F2a-precontracted aortic ring preparations of isolated rat aorta that contained endothelium. Relaxation by PRE did not occur in specimens without endothelium, and was inhibited by pretreatment with 10(-4) M NG-nitro-1-arginine methyl ester. Paeoniflorin and paeonol, the main active components of Paeoniae Radix, lacked a vasodilator effect. The effect of the component gallotannin was examined after treating PRE with tannase, but the product lacked a vasodilator effect. Pentagalloylglucose, hexagalloylglucose, heptagalloylglucose, and octagalloylglucose were extracted from PRE; they relaxed aortic rings with endothelium, but failed to relax aortic rings without endothelium. We conclude that PRE exhibits an endothelium-dependent vasodilator effect on isolated rat aorta. The active component is gallotannin.

Acetophenones↗