Search PubMed⌕ Search

Biomedical subjects

Y Gotoh

Publications and source records attributed to Y Gotoh.

At least 91 records · Page 5Linked to original sources
PubMed1996

Chemical modification of silk fibroin with N-acetyl-chito-oligosaccharides.

N-Acetyl-chito-oligosaccharides (NACOS)-silk fibroin (SF) conjugates (NACOS-CY-SF) were prepared by the reaction of solubilized SF and cyanuric chloride (CY)-activated NACOS modifier (NACOS-CY). N-Acetyl-D-glucosamine (NAG), as a model compound, was reacted with CY to clarify the chemical structure of the modifier. The 1H- and 13C-NMR spectra of the reactant suggest that the anomeric hydroxyl group of NACOS reacted with the chlorine atom of CY. The content of NACOS in the NACOS-CY-SF conjugates was calculated by comparing the integral values of the signals in the 1H-NMR spectra of the conjugates and the mixture of NACOS and SF. As the 1H-NMR spectrum of the conjugates showed a downfield shift of the aromatic protons of the tyrosine residue, the tyrosine residue in SF reacted with another chlorine atom of the triazine ring of the modifier. The result of the amino acid analysis of the conjugates suggests that lysine residues also reacted with the modifier.

Acetylglucosamine↗
PubMed1996

Chemical modification of the arginyl residue in silk fibroin: 2. Reaction of 1,2-cyclohexanedione in aqueous alkaline medium.

The arginyl residue of solubilized silk fibroin was chemically modified with 1,2-cyclohexanedione in aqueous alkaline medium to form a stable imidazolidinone ring, and its positive charge was masked. CD spectra of the modified silk fibroin in aqueous solution showed an increase in the fraction of random coil conformation. The increase may be caused by the exposure to alkaline medium in the modification reaction. FT-IR and CD spectra of the silk fibroin films before and after the modification indicated that the conformational change in the modified silk fibroin in the solid state did not occur by the modification of its arginyl residue with 1,2-cyclohexanedione. The chemical stability of the modified silk fibroin film was investigated in vitro with phosphate-buffered saline solution. The modified arginyl residue in the film was stable in the phosphate-buffered solution.

Animals↗
PubMed1996

Lingual factors enhance the increase of ornithine decarboxylase activity in rat jejunal mucosa after feeding.

Luminal nutrients are the main factors that stimulate ornithine decarboxylase (ODC) activity in rat intestinal mucosa following feeding. The aim of the present study was to determine whether lingual (oral) factors are related to the increase in jejunal ODC activity after feeding. ODC activity in the jejunum and liver was measured 3 hours after refeeding of 48-hour fasted rats. In the first experiment, rats were refed with a regular pellet, powder, or liquid diet. In the second experiment, rats were infused with the liquid diet through a gastric infusion tube following 48 hours' fasting. In the third experiment, the experimental rats had a gastric fistula that allowed free drainage from the stomach of all ingested liquid diet. In the fourth experiment, a truncal vagotomy was performed 1 week before the experiment. The increase of ODC activity in the jejunum of rats fed with the liquid diet was less than that of rats fed with the pellet diet or powder diet. The increase of ODC activity in the jejunal mucosa of rats infused through the gastric tube was less than that of rats fed per os, and the increase of ODC activity in the liver did not differ between these experimental groups. ODC activity did not increase in rats with a gastric fistula. Vagotomy did not affect the increase of jejunal ODC activity after feeding. In conclusion, the increase of ODC activity after feeding was attenuated in rats in which the diet was given by bypassing the mouth. This indicates that lingual factors enhance the increase of ODC activity in the jejunal mucosa after feeding, but the lingual factors alone do not increase ODC activity in the jejunum.

Animal Feed↗
PubMed1996

Efficient promoter cassettes for enhanced expression of foreign genes in dicotyledonous and monocotyledonous plants.

A series of chimeric promoters for higher-level expression of foreign genes in plants was constructed as fusions of a gene for beta-glucuronidase (GUS) with the terminator of a gene for nopaline synthase (nos) or of the cauliflower mosaic virus (CaMV) 35S transcript, and the strength of these promoters was assayed in transient and stable expression systems in tobacco and rice. As parts of these promoters, the CaMV 35S core promoter, three different 5'-upstream sequences of the 35S promoter, the first intron of a gene for phaseolin, and a 5'-untranslated sequence (omega sequence) of tobacco mosaic virus were used in various combinations. In tobacco and rice protoplasts, all three fragments of the 35S promoter (-419 to -90, -390 to -90 and -290 to -90, relative to the site of initiation of transcription), the intron, and the omega sequence effectively enhanced GUS activity. Some chimeric promoters allowed levels of GUS activity that were 20- to 70-fold higher than those obtained with the 35S promoter in pBI221. In tobacco protoplasts, the two longer fragments of the 35S promoter were more effective than the shortest fragment. In rice cells, by contrast, the shortest fragment was as effective as the two longer ones. The terminator of the 35S transcript was more effective than that of the nos gene for gene expression. In transgenic tobacco plants, a representative powerful promoter, as compared to the 35S promoter, allowed 10- and 50-fold higher levels of expression on average and at most, respectively, with no clear qualitative differences in tissue- and organ-specific patterns of expression. When the representative promoter was introduced into tobacco with a gene for luciferase, the autofluorescence of detached leaves after a supply of luciferin to petioles was great and was easily detectable by the naked eye in a dark room.

Amino Acid Oxidoreductases↗
PubMed1996

Necrotizing fasciitis due to group A streptococci: a clinicopathological study of six patients.

The recent worldwide appearance of invasive group A streptococcal infections has again called attention to streptococcal necrotizing fasciitis. However, in contrast to polymicrobial necrotizing fasciitis, the streptococcal form has not been thoroughly studied clinically. The objective of the study was to elucidate the characteristic features of recent cases of necrotizing fasciitis due exclusively to pure group A streptococci. We encountered six patients with these criteria at a single hospital in Japan during the last 12 years. A clinicopathological analysis was performed in these six patients. In three patients, the clinical signs and the laboratory findings were characteristic of systemic toxicity. In this group, the clinical presentation was a pale or blue-gray lesion associated with severe intravascular coagulation histologically involving the vessels in the lesion. In the three patients without signs of systemic toxicity, a swollen, erythematous skin lesion persisted for as long as one week; histologically, the intravascular coagulation within these lesions was mild. In clinicopathological terms, the entity in these six patients could be clearly classified as either fulminant or subacute. In the fulminant type, immediate surgical debridement of necrotic fascia is required; in the subacute type, incision and drainage alone are sufficient.

Adult↗
PubMed1996

Metanephric adenoma of the kidney: ultrastructural, immunohistochemical and lectin histochemical studies.

A case of metanephric adenoma, a rare benign tumor of the kidney is reported, and the results of ultrastructural, immunohistochemical, and lectin histochemical studies are presented. The patient was a 47 year old Japanese man presenting with a left renal tumor by ultrasonography. The nephrectomy material revealed a well-demarcated tumor with medullary appearance, and on histological examination the tumor was composed of cuboidal cells forming regular small tubules with hyalinous stroma. Incomplete glomeruloid structures were formed in a few elongated tubules composed of columnar cells. Metanephric blastema was not observed in the tumor or in the normal kidney. By electron microscope, the tumor cells were observed to be immature cells with microvilli at the apical surfaces. The neoplastic tubules were surrounded by basal lamina. The tumor cells were immunoreactive for Leu 7, epithelial membrane antigen, vimentin, and low molecular weight cytokeratin, and were weakly immunoreactive for S-100 protein. Peanut agglutinin, soybean agglutinin, and Dolichos biflorus agglutinin were bound to the apical surfaces of the tumor cells.

Adenoma↗
PubMed1996

The fission yeast pmk1+ gene encodes a novel mitogen-activated protein kinase homolog which regulates cell integrity and functions coordinately with the protein kinase C pathway.

We have isolated a gene, pmk1+, a third mitogen-activated protein kinase (MAPK) gene homolog from the fission yeast Schizosaccharomyces pombe. The predicted amino acid sequence shows the most homology (63 to 65% identity) to those of budding yeast Saccharomyces Mpk1 and Candida Mkc1. The Pmk1 protein contains phosphorylated tyrosines, and the level of tyrosine phosphorylation was increased in the dsp1 mutant which lacks an attenuating phosphatase for Pmk1. The level of tyrosine phosphorylation appears constant during hypotonic or heat shock treatment. The cells with pmk1 deleted (delta pmk1) are viable but show various defective phenotypes, including cell wall weakness, abnormal cell shape, a cytokinesis defect, and altered sensitivities to cations, such as hypersensitivity to potassium and resistance to sodium. Consistent with a high degree of conservation of amino acid sequence, multicopy plasmids containing the MPK1 gene rescued the defective phenotypes of the delta pmk1 mutant. The frog MAPK gene also suppressed the pmk1 disruptant. The results of genetic analysis indicated that Pmk1 lies on a novel MAPK pathway which does not overlap functionally with the other two MAPK pathways, the Spk1-dependent mating signal pathway and Sty1/Spc1/Phh1-dependent stress-sensing pathway. In Saccharomyces cerevisiae, Mpk1 is involved in cell wall integrity and functions downstream of the protein kinase C homolog. In contrast, in S. pombe, Pmk1 may not act in a linear manner with respect to fission yeast protein kinase C homologs. Interestingly, however, these two pathways are not independent; instead, they regulate cell integrity in a coordinate manner.

Amino Acid Sequence↗
PubMed1996

Nucleotide sequence of the gene encoding pepstatin-insensitive acid protease B, scytalidopepsin B, of Scytalidium lignicolum.

A chromosomal DNA fragment of Scytalidium lignicolum that encodes the mature enzyme region of acid protease B (Scytalidopepsin B), was cloned and its nucleotides sequenced. The fragment contained a 76-bp intron at the middle of the mature enzyme-coding region. The mature enzyme was composed of 206 amino acid residues with a molecular weight of 21,550. There were some discrepancies between the amino acid sequence deduced from these results and that previously established by protein sequencing.

Amino Acid Sequence↗
PubMed1996

[Analysis of locally controlled esophageal carcinomas treated with radiotherapy].

Of 227 esophageal carcinomas treated with a radiation dose of 60 Gy or more, 100 patients had no tumor or ulceration (with or without stenosis) of the esophagus after irradiation. We analyzed local control factors of these 100 patients to determine the need for further treatment. The cumulative local control rate at five years was 40% in all cases, 37% in 21 cases without any stenosis of the esophagus and 40% in 79 cases with stenosis. The presence of stenosis of the esophagus after irradiation was not a critical factor in predicting final local control. Local recurrence of tumors with findings of Borrmann III or Borrmann IV by the pretreatment esophageal barium study, tumors controlled after a total dose of more than 80 Gy, tumors without low dose rate telecobalt therapy (LDRT; 1 Gy/hour, 5 to 7Gy/day, a total dose of 12 to 15 Gy) as boost therapy, and apparently controlled tumors with a stenotic ratio of 60% or more or with 5 cm or more length of stenosis of the esophagus after irradiation was significantly higher than that of the others (p < 0.05). Multivariate analysis revealed that findings of pretreatment barium study, total dose, with or without LDRT, and length of stenosis of the esophagus after irradiation were significantly important factors in local control. Members of the high risk group of apparently controlled tumors should undertake surgical treatment or further intensive chemotherapy.

Aged↗
PubMed1995

Activation of the estrogen receptor through phosphorylation by mitogen-activated protein kinase.

The phosphorylation of the human estrogen receptor (ER) serine residue at position 118 is required for full activity of the ER activation function 1 (AF-1). This Ser118 is phosphorylated by mitogen-activated protein kinase (MAPK) in vitro and in cells treated with epidermal growth factor (EGF) and insulin-like growth factor (IGF) in vivo. Overexpression of MAPK kinase (MAPKK) or of the guanine nucleotide binding protein Ras, both of which activate MAPK, enhanced estrogen-induced and antiestrogen (tamoxifen)-induced transcriptional activity of wild-type ER, but not that of a mutant ER with an alanine in place of Ser118. Thus, the activity of the amino-terminal AF-1 of the ER is modulated by the phosphorylation of Ser118 through the Ras-MAPK cascade of the growth factor signaling pathways.

Amino Acid Sequence↗
PubMed1995

Activation of two isoforms of mitogen-activated protein kinase kinase in response to epidermal growth factor and nerve growth factor.

Mitogen-activated protein kinase kinase (MAPKK) is a dual-specificity protein kinase which phosphorylates and activates mitogen-activated protein kinase (MAPK). cDNAs encoding two isoforms of MAPKK, MAPKK1 and MAPKK2 (also known as MEK1 and MEK2), have been cloned in mammalian cells. To analyze the characteristics of MAPKK1 and MAPKK2 individually, we have produced specific anti-MAPKK serum against each isoform. MAPKK1 and MAPKK2 have apparent molecular masses of 45 kDa and 47 kDa, respectively, on SDS/polyacrylamide gel electrophoresis. In mouse tissues, MAPKK1 was highly enriched in brain, while MAPKK2 was present relatively evenly. In rat fibroblastic 3Y1 cells, epidermal growth factor (EGF) treatment induced activation of both MAPKK1 and MAPKK2. Immunoprecipitation experiments have shown that the time courses of activation and deactivation of both isoforms of MAPKK were superimposed. In PC12 cells, both MAPKK1 and MAPKK2 were activated in response to nerve growth factor (NGF) as well as EGF, and the time courses of activation and deactivation of both isoforms were indistinguishable from each other in the NGF-stimulated cells and also in the EGF-stimulated cells. Furthermore, localization of both MAPKK1 and MAPKK2 in the cytoplasm was unchanged in response to EGF and NGF. Thus, the same or quite similar mechanisms may operate in the regulation of the activation and deactivation of two isoforms of MAPKK, and both kinases might have redundant functions when expressed in the same cell.

Animals↗
PubMed1995

Initiation of Xenopus oocyte maturation by activation of the mitogen-activated protein kinase cascade.

Mitogen-activated protein kinase (MAPK) and MAPK kinase (MAPKK) are activated during Xenopus oocyte maturation concomitant with the activation of maturation promoting factor (MPF). We reported previously that an anti-MAPKK neutralizing antibody inhibited progesterone- or Mos- induced initiation of oocyte maturation. Here, we show that the injection of CL100 (also called MAPK phosphatase-1) into immature oocytes inhibited progesterone-induced oocyte maturation as well as MAPK activation and that injection of mRNA encoding a constitutively active MAPKK induced activation of histone H1 kinase and germinal vesicle breakdown in the absence of progesterone. Injection of recombinant STE11 protein (a yeast MAPKK kinase) also induced initiation of oocyte maturation. These data support the idea that the MAPKK/MAPK cascade plays an important role in oocyte maturation. Interestingly, injection of the active MAPKK mRNA or the STE11 protein resulted in induction and accumulation of Mos protein. Furthermore, in the presence of cycloheximide, the STE11-induced activation of MPF as well as the induction and accumulation of Mos was blocked, and the activation of MAPK was greatly reduced. The increase in Mos protein and the activation of MAPK by injecting cyclin A protein into immature oocytes were both blocked also by cycloheximide treatment. These results are consistent with an idea that there may exist a positive feedback loop consisting of Mos, the MAPKK/MAPK cascade, and MPF, which may be important for the initiation of oocyte maturation induced by progesterone.

Animals↗
PubMed1995

Induction of neurite outgrowth by MAP kinase in PC12 cells.

Treatment of PC12 cells with nerve growth factor (NGF) results in neural differentiation of the cells, inducing neurite outgrowth. Ras protein has been shown to play an essential role in this process. To examine whether or not the MAP kinase (MAPK) cascade mediates the NGF- and Ras-induced neural differentiation process, we injected PC12 cells with constitutive active forms of each components of the MAPK cascade. When a moderately active mutant of Xenopus MAPK kinase (S222E-MAPKK) in which Ser 222 was changed into glutamic acid was injected, the neurite outgrowth of PC12 cells occurred to some extent. Injection of an N-terminal truncated STE11 protein (delta N-STE11), a constitutively active form of STE11 which is a yeast MAPKK kinase, induced neurite outgrowth in PC12 cells. Furthermore, injection of thiophosphorylated MAPK, but not purified active MAPK, into PC12 cells resulted in neurite outgrowth. Thiophosphorylated MAPK was resistant to protein phosphatase 2A treatment, while purified active MAPK was inactivated by this treatment. All these results have suggested that sustained activation of MAPK is sufficient for PC12 cell differentiation. In accord with this, the delta N-STE11- or S222E- MAPKK-induced neurite outgrowth was inhibited by coinjection of CL-100 protein, a dual-specificity phosphatase that is capable of inactivating MAPK.

Animals↗
PubMed1995

Evidence for multiple activators for stress-activated protein kinase/c-Jun amino-terminal kinases. Existence of novel activators.

Stress-activated protein kinases (SAPKs) or c-Jun amino-terminal kinases (JNKs), which belong to a subgroup of the mitogen-activated protein kinase (MAPK) superfamily, are activated in response to a variety of stresses in mammalian cells. An activity to activate a recombinant rat SAPK alpha was detected in extracts obtained from rat fibroblastic 3Y1 cells exposed to hyperosmolar media and was resolved into unadsorbed and adsorbed fractions on Q-Sepharose chromatography. The adsorbed activity was identified as XMEK2/SEK1/MKK4 by using several anti-XMEK2 antibodies. Thus, a 45-kDa protein that was recognized specifically by these anti-XMEK2 antibodies co-eluted with the SAPK alpha activating activity during chromatography on Q-Sepharose and Superose 6, and the activity could be immunoprecipitated by the antibodies from these fractions. The unadsorbed activity, whose level was much greater than that of the adsorbed activity, did not contain XMEK2/SEK1/MKK4 and was also activated in a time-dependent manner by osmotic shock. This activity was further resolved into several peaks during chromatography on heparin-Sepharose and hydroxylapatite. Most of these peaks eluted separately from major peaks of a kinase activity toward p38/MPK2, another subgroup of the MAPK superfamily, whereas the activated XMEK2/SEK1/MKK4 could phosphorylate p38/MPK2 efficiently. These results indicate the existence of multiple activators for SAPK/JNK; one is XMEK2/SEK1/MKK4, and the others are previously undescribed factors.

Amino Acid Sequence↗
PubMed1995

Involvement of the MAP kinase cascade in Xenopus mesoderm induction.

Mitogen-activated protein kinase (MAPK) is activated by MAPK kinase (MAPKK) in a variety of signaling pathways. This kinase cascade has been shown to function in cell proliferation and differentiation, but its role in early vertebrate development remains to be investigated. During early vertebrate embryogenesis, the induction and patterning of mesoderm are thought to be determined by signals from intercellular factors such as members of the fibroblast growth factor (FGF) family and members of the transforming growth factor-beta family. Here we show that the microinjection of either mRNA encoding a constitutively active mutant of MAPKK or mRNA encoding a constitutively active form of STE11, a MAPKK kinase, leads to the induction of mesoderm in ectodermal explants from Xenopus embryos. Moreover, the expression of MAPK phosphatase-1 (MKP-1, also called CL100) blocks the growth factor-stimulated mesoderm induction. Furthermore, injection of CL100 mRNA into two-cell stage embryos causes severe defects in gastrulation and posterior development. The effects induced by CL100 can be rescued by co-injection of wild-type MAPK mRNA. Thus, the MAPK cascade may play a crucial role in early vertebrate embryogenesis, especially during mesoderm induction.

Animals↗