Biomedical subjects

T Kono

Publications and source records attributed to T Kono.

At least 37 records · Page 2Linked to original sources

PubMed2001

Disruption of imprinted expression of U2afbp-rs/U2af1-rs1 gene in mouse parthenogenetic fetuses.

The present study shows that the U2afbp-rs gene, a paternally expressed imprinted gene, is activated and expressed in a biallelic manner from maternal alleles in parthenogenetic mouse fetuses on day 9.5 of gestation. The mean expression was detected to be 88% (31-134%) of that in control biparental fetuses, using real-time quantitative reverse transcription and polymerase chain reaction. This disrupted expression of the gene was associated with changes in the chromatin structure but not with the methylation pattern in the regulation region. The present results show that parental specific expression of imprinted genes is not always maintained in uniparental embryos. This suggests that both parental genomes are necessary to establish parental specific expression of the U2afbp-rs gene.

Animals↗

PubMed2001

Distinct roles for the N- and C-terminal regions in the cytotoxicity of pierisin-1, a putative ADP-ribosylating toxin from cabbage butterfly, against mammalian cells.

Pierisin-1 is an 850-aa cytotoxic protein found in the cabbage butterfly, Pieris rapae, and has been suggested to consist of an N-terminal region with ADP-ribosyltransferase domain and of a C-terminal region that might have a receptor-binding domain. To elucidate the role of each region, we investigated the functions of various fragments of pierisin-1. In vitro expressed polypeptide consisting of amino acid residues 1-233 or 234-850 of pierisin-1 alone did not show cytotoxicity against human cervical carcinoma HeLa cells. However, the presence of both polypeptides in the culture medium showed some of the original cytotoxic activity. Introduction of the N-terminal polypeptide alone by electroporation also induced cell death in HeLa cells, and even in the mouse melanoma MEB4 cells insensitive to pierisin-1. Thus, the N-terminal region has a principal role in the cytotoxicity of pierisin-1 inside mammalian cells. Analyses of incorporated pierisin-1 indicated that the entire protein, regardless of whether it consisted of a single polypeptide or two separate N- and C-terminal polypeptides, was incorporated into HeLa cells. However, neither of the terminal polypeptides was incorporated when each polypeptide was present separately. These findings indicate that the C-terminal region is important for the incorporation of pierisin-1. Moreover, presence of receptor for pierisin-1 in the lipid fraction of cell membrane was suggested. The cytotoxic effects of pierisin-1 were enhanced by previous treatment with trypsin, producing "nicked" pierisin-1. Generation of the N-terminal fragment in HeLa cells was detected after application of intact entire molecule of pierisin-1. From the above observations, it is suggested that after incorporation of pierisin-1 into the cell by interaction of its C-terminal region with the receptor in the cell membrane, the entire protein is cleaved into the N- and C-terminal fragments with intracellular protease, and the N-terminal fragment then exhibits cytotoxicity.

ADP Ribose Transferases↗

PubMed2001

A retrospective study looking at the long-term complications of Q-switched ruby laser in the treatment of nevus of Ota.

BACKGROUND AND OBJECTIVE: Despite the extensive use of QS Ruby, there is no report looking at its long-term complication in the treatment of nevus of Ota. To look at the long-term complications of nevus of Ota patients treated with QS Ruby laser. STUDY DESIGN/MATERIALS AND METHODS: A teaching hospital in Japan where over 400 patients with nevus of Ota had been treated since 1984. Hundred and one nevus of Ota patients that had been treated with QS Ruby laser, but had not received treatment for 12 months prior to the study, were called back for further assessment. Patients were called back to the hospital where they were interviewed and examined by two independent clinicians. RESULTS: Hypopigmentation was the most common complication, affecting 16.8% of the patients and 5.9% had hyperpigmentation. One patient that had complete clearance post-laser surgery developed recurrence. CONCLUSION: QS Ruby laser is effective in the treatment of nevus of Ota. Recurrence is rare, but hypopigmentation is common and can be permanent. Further prospective study comparing its use with other Q-switched lasers is necessary.

Female↗

PubMed2001

The analysis of expressed genes in the kidney of Japanese flounder, Paralichthys olivaceus, injected with the immunostimulant peptidoglycan.

Immunostimulants are widely used in aquaculture, but there are few reports on the genes that are expressed by their stimulation. Therefore, in this study, expressed genes in the kidney of Japanese flounder Paralichthys olivaceus injected with the immunostimulant peptidoglycan were analysed. The results of single-pass sequencing of ESTs from 198 clones (AU090255-AU090451, AU090935) from kidney cDNA are presented. Sequences of the cDNA clones were compared with sequences in the GenBank database. One hundred and six clones (53.5%) appeared to be completely unknown and are likely to represent newly described genes, whereas 92 clones (46.5%) were identified based on matches to sequences in the database. The results contain the genes such as alpha globin (AU090287), several ribosomal proteins (AU090-263, 274, 299, 351, 365, 375, 377, 382, 434, 445), heat shock protein 90 (AU090374) and cytochrome oxidase subunit (AU090385). Immune related cDNAs identified from the kidney were immunoglobulin heavy (AU090291) and light chain (AU090352), beta2-microglobulin (AU090280), macrophage inflammatory protein 1-alpha precursor (AU090535), thymosin beta-10 (AU090391), lysozyme (AU090322) and MHC class IIalpha (AU090435). It is possible that expression of macrophage inflammatory protein 1-alpha results in macrophage activation as a consequence of peptidoglycan treatment.

Amino Acid Sequence↗

PubMed2001

Coronary arteriovenous fistula with annuloaortic ectasia and aortic regurgitation.

Cardiac catheterization in a 57-year-old man disclosed a left coronary artery-coronary sinus fistula with annuloaortic ectasia and severe aortic regurgitation. During surgery, the fistula vessel was exposed and clamped before infusing cardioplegic solution for cardiac arrest. The proximal orifice of the fistula was closed directly and the distal orifice doubly ligated. A modified Bentall's procedure was conducted without difficulty. The postoperative course was uneventful.

Aortic Valve Insufficiency↗

PubMed2001

CT findings of descending necrotising mediastinitis via the carotid space ('Lincoln Highway').

A 3-year-old girl with fever and neck swelling showed widening of the superior mediastinum on chest radiographs. Contrast-enhanced CT of the neck revealed ill-defined low-attenuation areas with a thick, enhanced rim adjacent to the hypertrophied palatine tonsil. The abscess extended inferiorly within the carotid sheath between the carotid artery and the internal jugular vein into the anterior mediastinum. The carotid space is considered an important conduit of descending necrotising mediastinitis and is called the 'Lincoln Highway' as previously suggested.

Carotid Arteries↗

PubMed2001

Adaptive cytoprotection against acetic acid induced colonic injury in rats.

BACKGROUND AND AIMS: The phenomenon of prostaglandin dependent adaptive cytoprotection has been well established in the stomach and duodenum but not in the colon. This study investigated whether it also occurs in the colon. METHODS: Fisher rats received intracolonic administration (0.5 ml) of saline or acetic acid at low concentrations (0.01-5%) followed by high concentration (25%) at various intervals (10-720 min). The distal colon was removed 2 h after acetic acid (25%), and colonic injury was assessed macroscopically and histologically and scored. Indomethacin (5 mg/kg) or N(G)-nitro-L-arginine methyl ester (10 mg/kg) was injected intraperitoneally 1 h or 30 min before pretreatment with 1% acetic acid. RESULTS: Acetic acid (25%) administered into the colon induced 11.6+/-0.2 macroscopic scores and 10.0+/-0.4 histological scores in saline-pretreated rats, which were reduced to 0.3+/-0.2 and 1.8+/-0.5 by acetic acid (1%) pretreatment, respectively. The protective effect assessed macroscopically and histologically was dose related (0.01-1% acetic acid) and long lasting (maximal at 30 min and lasting up to 6 h). Indomethacin abolished the colonic adaptive cytoprotection while nitric oxide synthase inhibitor had no effect. CONCLUSIONS: These data show that prostaglandin dependent adaptive protection is induced by exposure of the colon to low concentrations of acetic acid, suggesting that endogenous defense mechanisms are increased in the colon by mild irritants.

Acetic Acid↗

PubMed2001

Intestinointestinal inhibitory reflexes: effect of distension on intestinal slow waves.

The motor response induced by intraluminal distension of the small intestine has been well investigated. However, little is known of the myoelectrical response to intraluminal distension. The aim of this study was to investigate the effects of oral- and anal-side distension on jejunal slow waves in dogs. The study was performed in 10 healthy female hound dogs implanted with three pairs of electrodes on the jejunum and an intestinal fistula. The first study session was designed to investigate the effects of anal-side distension on jejunal myoelectrical activity in fasting state. The protocol consisted of a 30-min baseline recording, a 30-min recording during anal-side balloon distension located 5 cm distal to the most distal pair of electrodes, and another 30-min recording after distension. The second session was designed to investigate the effect of oral-side distension with the balloon 5 cm proximal to the most proximal pair of electrodes. Jejunal slow waves were severely impaired by both anal- and oral-side distension. The dominant power was significantly reduced from -2.96 +/- 0.90 dB at baseline to -6.00 +/- 0.64 dB during anal-side distension (P < 0.0005) and from -3.90 +/- 0.85 dB at baseline to -7.17 +/- 0.90 dB during oral-side distension (P < 0.001). The percentage of normal 17 to 22-cpm slow waves was significantly decreased from 97.39 +/- 0.88% to 83.48 +/- 3.12% during anal-side distension (P < 0.0005) and from 92.49 +/- 2.42% to 68.80 +/- 7.24% during oral-side distension (P < 0.002). The percentage of slow wave coupling was decreased from 95.08 +/- 2.27% to 52.48 +/- 7.73% during anal-side distension (P < 0.0005) and from 84.82 +/- 6.75% to 49.21 +/- 8.91% during oral-side distension (P < 0.001). The instability coefficient of the dominant frequency was significantly increased during anal-side distension. In conclusion, intraluminal distension of the jejunum severely impairs jejunal slow waves. The slow waves on both sides of distension become less coupled, less regular, and are of lower amplitude.

Animals↗

PubMed2001

Granulomatous slack skin: an ultrastructural study.

BACKGROUND: Granulomatous slack skin is a rare lymphoproliferative disorder characterized clinically by gradual development of pendulous folds of lax erythematous skin in flexural areas, and histologically by non-necrotizing granuloma, with numerous multinucleated giant cells, mononuclear histiocytes, and atypical lymphocytes associated with loss of elastic fibers. Although there are many reports describing the histological and immunophenotypic features of this disease, only a few have described the ultrastructural features. METHODS: Here we report a case of granulomatous slack skin and describe the ultrastructural findings. RESULTS AND CONCLUSION: We could detect some previously not noted abnormal findings in multinucleated giant cells, histiocytes, and atypical lymphocytes.

Adult↗

PubMed2001

Combined use of normal mode and Q-switched ruby lasers in the treatment of congenital melanocytic naevi.

Melanocytic naevi are common and they are often removed for cosmetic purposes. In recent years, both the normal mode ruby laser and the Q-switched ruby laser have been used, either alone or in combination, for the removal of congenital melanocytic naevi. However, their success was restricted to partial lightening and incomplete histological clearance. We report a new approach, with combined use of normal mode and Q-switched ruby lasers, that improves the effectiveness of the laser removal of pigmentation in congenital melanocytic naevi.

Adult↗

PubMed2001

Identification of a new imprinted gene, Rian, on mouse chromosome 12 by fluorescent differential display screening.

Systematic screening of differentially expressed genes among androgenetic, parthenogenetic, and normal embryos by means of fluorescent differential display revealed five imprinted genes. One of them, named Rian, was expressed exclusively from the maternal allele and was closely linked to an imprinted gene, Meg3(Gtl2), mapped to the distal end of chromosome 12. The Rian transcript did not have any apparent open reading frame, and its transcript was exclusively localized to the nucleus.

Animals↗

PubMed2001

Cloned mice from fetal fibroblast cells arrested at metaphase by a serial nuclear transfer.

Cloning using G(0)-arrested somatic cells has led to the suggestion that this stage of the cell cycle is necessary for the success of cloning. In this study we report that cloned mice can be generated from fetal fibroblasts arrested at metaphase of the cell cycle. The procedure involves fusing a metaphase-arrested fetal fibroblast to an enucleated oocyte. After parthenogenetic activation a polar body and single diploid pronucleus were formed. Some of these were allowed to develop to the blastocyst stage, while others were enucleated and the nucleus was transferred to an enucleated fertilized 1-cell embryo. After the single transfer technique, 2 out of 164 developed to late stages of gestation were dead with gross abnormalities. However, after the serial nuclear transfer, 5 out of 272 embryos were recovered live at Day 19.5, and 2 of these went on to develop into apparently normal adults. All of the cloned embryos showed severe placental hypertrophy and defective differentiation of placental tissues. This study illustrates that reprogramming can occur after nuclear transfer at metaphase of the cell cycle.

Animals↗

PubMed2001

Effect of ecabet sodium enema on mildly to moderately active ulcerative proctosigmoiditis: an open-label study.

OBJECTIVES: Ecabet sodium (ES), a nonabsorbable antigastric ulcer agent, has been shown to adhere to the region of an ulcer. It topically enhances gastric mucosal defensive factors such as the endogenous prostaglandins, capsaicin-sensitive sensory nerves, nitric oxide, and mucin. All of these mucosal defensive factors play an important role in maintaining the mucosal integrity of the colon and rectum. Therefore, we investigated the effect of ES in patients with mildly to moderately active ulcerative proctosigmoiditis. METHODS: In an open-label study, seven patients with mildly to moderately active ulcerative colitis (UC) who had an inflamed mucosa in the rectum and/or sigmoid and were resistant to 4-wk topical and systemic standard treatment were treated with an ES enema b.i.d. for 14 days. The enema consisted of ES (1 g) and tepid water (20 or 50 ml). These patients were assessed by the Clinical Activity Index, colonoscopically, and histologically before and after the ES therapy. The ES therapy was started after obtaining informed consent from the patients. RESULTS: Six of the seven patients responded to therapy and achieved clinical, endoscopic, and histological remissions. One patient was withdrawn because of increased stool frequency. All six patients who completed the study showed a significant change in the mean Clinical Activity Index score from 5.3+/-1.4 (mean +/- SD) to 0.5+/-0.8 (p < 0.05), in the colonoscopic score from 3.0+/-0.9 to 0.8+/-0.4 (p < 0.05), and in the histological score from 2.7+/-0.5 to 0.5+/-0.6 (p < 0.05), and achieved remission at the end of the study. There were no side effects attributable to the ES therapy. Five of the six patients are still in clinical remission after a median follow-up period of 5 months. CONCLUSIONS: The ES enemas proved to be a safe and potentially useful adjuvant therapy currently available for treating patients with mildly to moderately active ulcerative proctosigmoiditis. A controlled study is necessary to confirm our results.

Abietanes↗

PubMed2001

Glucose uptake and metabolism by cultured human skeletal muscle cells: rate-limiting steps.

To use primary cultures of human skeletal muscle cells to establish defects in glucose metabolism that underlie clinical insulin resistance, it is necessary to define the rate-determining steps in glucose metabolism and to improve the insulin response attained in previous studies. We modified experimental conditions to achieve an insulin effect on 3-O-methylglucose transport that was more than twofold over basal. Glucose phosphorylation by hexokinase limits glucose metabolism in these cells, because the apparent Michaelis-Menten constant of coupled glucose transport and phosphorylation is intermediate between that of transport and that of the hexokinase and because rates of 2-deoxyglucose uptake and phosphorylation are less than those of glucose. The latter reflects a preference of hexokinase for glucose over 2-deoxyglucose. Cellular NAD(P)H autofluorescence, measured using two-photon excitation microscopy, is both sensitive to insulin and indicative of additional distal control steps in glucose metabolism. Whereas the predominant effect of insulin in human skeletal muscle cells is to enhance glucose transport, phosphorylation, and steps beyond, it also determines the overall rate of glucose metabolism.

3-O-Methylglucose↗

PubMed2001

Development and characterization of 1C6-203, a new monoclonal antibody specific to human thymidine phosphorylase.

Thymidine phosphorylase (dThdPase) is an essential enzyme for activation of the oral cytostatic drug capecitabine and its intermediate metabolite, doxifluridine, to 5-fluorouracil in tumors. Methods to estimate dThdPase expression in tumor tissue might be useful to predict the efficacy of capecitabine and doxifluridine in cancer patients. We established a new monoclonal antibody (MAb), 1C6-203, applicable for dThdPase immunohistochemistry and compared its staining characteristics with those of a previously established MAb, 654-1. In 4% paraformaldehyde-fixed colorectal carcinoma, 1C6-203 and 654-1 stained cancer cells in 19/30 and 9/30 patients, respectively. In 10% formalin-fixed colorectal carcinoma, 1C6-203 and 654-1 stained cancer cells in 18/30 and 6/30 patients, respectively. In negative 10% formalin-fixed tissues, microwave treatment improved the positivity of 654-1-stained cancer cells. These results suggest that an epitope recognized by 1C6-203 is resistant to epitope masking by formaldehyde fixation, whereas that for MAb 654-1 is sensitive. Therefore, MAb 1C6-203 might be more suitable than MAb 654-1 for evaluating dThdPase expression in colorectal carcinoma.

Animals↗

PubMed2001

Computed tomographic evaluation of the position of the leg for mortise radiographs.

We investigated the most advantageous internal rotation angle of the leg for mortise radiographs. One hundred and twenty-eight feet of 64 healthy volunteers with no histories of ankle or foot pathology (72 feet of 36 males, 56 feet of 28 females) were examined. The subjects had an average age of 29 years (range, 19 to 51 years), average height of 167 cm (range, 157 to 181 cm), and average foot length of 25 cm (range, 23 to 27 cm). We obtained a plain axial view at the level of the central patella and 5 mm proximal to the tibial plafond using computed tomography, and investigated the inclination angle of the distal tibiofibular joint to a horizontal plane, regarding it as a mortise angle. The mean mortise angle was 19.1 +/- 5.0 degrees. However, two peaks were observed at around 15 degrees and 20 degrees. This indicated that the mean mortise angle of the males was 21.2 +/- 4.6 degrees, and the mean mortise angle of the females was 16.4 +/- 4.1degrees, with a significant difference between the males and the females' mortise angle (P < 0.0001). There was no correlation between the mortise angle and the height (P = 0.899 in the males, and P = 0.871 in the females), nor between the mortise angle and the foot length (P = 0.359 in the males, and P = 0.512 in the females). Therefore, we concluded that the internal rotation angle of the leg for mortise radiography should be generally set up at about 20 degrees on males and 15 degrees on females.

Adult↗

PubMed2001

Production of cloned mice from embryonic stem cells arrested at metaphase.

In mammals, cloned individuals can be produced from somatic cells. The combined use of gene targeting in embryonic stem cells and cloning contributes to the investigation of gene function in mammals. However, one of the major limitations to cloning is the low viability of cloned embryos, leading typically to high rates of pre- and postnatal death. The present study investigated whether cloning efficiency is influenced by the procedural differences involved in using transfected embryonic stem cells arrested at M phase for cloning by both single and serial transfer. In contrast to a previous study, in which fibroblasts were used, in the present study using embryonic stem cells there was no difference in the rate of production of cloned pups after the use of a single or serial nuclear transfer, although the proportion of blastocysts (70% versus 51%) was significantly higher (P < 0.001) after serial nuclear transfer. After embryo transfer of 445 blastocysts, 218 (49%) implanted and 27 (6% of blastocysts transferred) live pups were born. Of these 27 pups, 23 developed to adults of apparently normal fertility. Of these adults, 39% (n = 9) were derived from targeted embryonic stem cells, which is similar to the proportion of targeted embryonic stem cells in the population used for cloning. This study showed that cloning with embryonic stem cells is a viable procedure resulting in the production of transgenic cloned adults.

Animals↗

PubMed2001

Human fibroblasts ubiquitously express glutamic acid decarboxylase 65 (GAD 65): possible effects of connective tissue inflammation on GAD antibody titer.

BACKGROUND: Type 1 diabetes is caused by a destruction of pancreatic beta cells due to autoimmunity. Autoantibody against glutamic acid decarboxylase (GAD) 65 expressed in pancreatic beta cells is widely used as a predictive marker for pancreatic destruction. In this study, we hypothesized that if certain cells in periodontal tissues could express GAD, then it may influence GAD antibody titer. METHODS: We used: 1) reverse transcription-polymerase chain reaction (PCR) analysis to detect GAD 65 mRNA in various cells; 2) nucleotide sequencing analysis to confirm that amplified PCR product is the gene encoding GAD; and 3) Western blotting to determine the expression of GAD 65 protein in human gingival fibroblasts. Immunohistochemical staining of GAD 65 protein in normal and inflamed gingiva was performed to examine the potential influence of periodontal inflammation on GAD 65 expression. GAD antibody titer in sera of periodontal patients as well as healthy subjects was measured to determine if periodontal patients could develop autoantibody against GAD 65. RESULTS: Cultured human gingival, periodontal, and dermal fibroblasts and mesangial cells expressed GAD mRNA. Nucleotide sequencing analyses confirmed the amplified PCR product as GAD 65. Western immunoblotting analyses and immunohistochemical staining revealed that the GAD 65 protein was expressed in vitro and in vivo. The expression of GAD 65 in inflamed tissue was higher than that in normal tissues. Two of 62 periodontal patients without diabetes showed an increased antibody titer against GAD 65, while none of the systemically healthy subjects showed an increased antibody titer against this antigen. CONCLUSIONS: We concluded that periodontal inflammation may result in higher levels of GAD and influence GAD antibody titer, and, hence, affect diabetic diagnosis based upon GAD antibody production.

Actins↗