Biomedical subjects

T Endo

Publications and source records attributed to T Endo.

At least 235 records · Page 13Linked to original sources

PubMed1999

The role of chloroplastic NAD(P)H dehydrogenase in photoprotection.

After a brief exposure to supra-saturating light, leaves of a tobacco transformant, in which chloroplastic NAD(P)H dehydrogenase (NDH) was defective, showed more severe photoinhibition than the wild-type, when judged by the parameter of chlorophyll fluorescence Fv/Fm. Repeated application of supra-saturating light eventually resulted in chlorosis in the NDH-defective mutant, while the wild-type sustained less photodamage and was able to recover from it. The mechanism of the phenomena is discussed with respect to the potential role of NDH in photosynthesis.

Chloroplasts↗

PubMed1999

Age-related decrease in brain-derived neurotrophic factor gene expression in the brain of the zitter rat with genetic spongiform encephalopathy.

The zitter rat exhibits a progressive degradation in neuronal cells and genetic spongiform encephalopathy with age. In order to elucidate the involvement of the expression of the neurotrophic factor in neuropathology of the rat, we quantified mRNA levels of neurotrophic factors (nerve growth factor, brain-derived neurotrophic factor (BDNF), and neurotrophin-3, ciliary neurotrophic factor and glial-cell-line-derived neurotrophic factor) in the zitter rat brain. Expression of the BDNF gene was lower in the zitter rat brain (cerebrum, cerebellum, and brainstem regions). Interestingly, kinase activity of mitogen activated protein kinase (MAPK) Erk2 involved in the expression of BDNF was also down regulated, despite an unchanging expression of MAPK protein. These results show the possible involvement of a MAPK pathway in BDNF mRNA reduction in the zitter rat brain.

Aging↗

PubMed1999

Pharbin, a novel inositol polyphosphate 5-phosphatase, induces dendritic appearances in fibroblasts.

We have cloned a cDNA encoding a novel protein pharbin with a homology to inositol polyphosphate 5-phosphatases. Pharbin contains relatively well-conserved catalytic motifs for 5-phosphatase, a proline-rich sequence corresponding to the SH3-binding motif, and a sequence consistent with the CaaX motif at the C-terminus. COS-7 cells transfected with pharbin exhibited elevated hydrolytic activity on the 5-phosphate group of inositol 1,4,5-trisphosphate, inositol 1,3,4,5-tetrakisphosphate, and phosphatidylinositol 4, 5-bisphosphate. Thus, pharbin indeed serves as an inositol polyphosphate 5-phosphatase. When pharbin was transfected to C3H/10T1/2 fibroblasts, it was located to the plasma membrane-associated structures including membrane ruffles. The cells were converted to dendritic forms within 24 h. The protein with deleted or point-mutated CaaX motif hardly induced the dendritic forms but remained associated with the membranes. These results imply that the CaaX motif is required for the morphological alteration but that some other structural element is likely to also be responsible for the membrane localization.

Amino Acid Sequence↗

PubMed1999

Sec-dependent pathway and DeltapH-dependent pathway do not share a common translocation pore in thylakoidal protein transport.

Thylakoidal proteins of plant chloroplasts are transported to thylakoids via several different pathways, including the DeltapH-dependent and the Sec-dependent pathways. In this study, we asked if these two pathways utilize a common translocation pore. A fusion protein consisting of a 23-kDa subunit of the oxygen evolving complex and Escherichia coli biotin carboxyl carrier protein was biotinylated in E. coli cells and purified. When incubated with isolated pea thylakoids in the absence of avidin, the purified fusion protein was imported into the thylakoids via the DeltapH-dependent pathway. However in the presence of avidin, the fusion protein became lodged in the thylakoid membranes, with its N terminus reaching the thylakoidal lumen, while its C-terminal segment complexed with avidin exposed on the thylakoidal surface. The translocation intermediate of the fusion protein inhibited the import of authentic 23-kDa subunit, suggesting that it occupies a putative translocation pore for the DeltapH-dependent pathway. However the intermediate did not block import of the 33-kDa subunit of the oxygen evolving complex, which is a substrate for the Sec-dependent pathway. These results provide evidence against the possibility of a common translocation pore shared by the Sec-dependent pathway and the DeltapH-dependent pathway.

Acetyl-CoA Carboxylase↗

PubMed1999

Association of matrilysin expression with recurrence and poor prognosis in human esophageal squamous cell carcinoma.

Matrix metalloproteinase-7 (matrilysin) has been implicated in tumor invasion and metastasis as well as tumor initiation and growth. In this study, we analyzed an association between immunohistochemically detected matrilysin expression at the invasive front in esophageal squamous cell carcinomas and clinicopathological characteristics and determined whether matrilysin predicts recurrence and/or survival Matrilysin expression at the invasive front was detected in 49% of 100 carcinoma tissues and was associated with the depth of invasion (P < 0.0001), advanced tumor stage (P = 0.0159), recurrences (P = 0.0002), and recurrences within the first postoperative year (P = 0.002). Patients with matrilysin-positive carcinoma had a significantly shorter disease-free and overall survival time than did those with a matrilysin-negative one (P < 0.0001). Matrilysin remained a significant predictive value for disease-free and overall survival in multivariate analysis, including conventional clinicopathological factors (P = 0.0007 and 0.0004, respectively). Our results suggest that matrilysin may play a key role in the progression of esophageal carcinoma and that its detection may be useful for the prediction of recurrence and poor prognosis and, possibly, for selecting patients for anti-matrix metalloproteinase therapy.

Carcinoma, Squamous Cell↗

PubMed1999

FGF7 and FGF10 directly induce the apical ectodermal ridge in chick embryos.

During vertebrate limb development, the apical ectodermal ridge (AER) plays a vital role in both limb initiation and distal outgrowth of the limb bud. In the early chick embryo the prelimb bud mesoderm induces the AER in the overlying ectoderm. However, the direct inducer of the AER remains unknown. Here we report that FGF7 and FGF10, members of the fibroblast growth factor family, are the best candidates for the direct inducer of the AER. FGF7 induces an ectopic AER in the flank ectoderm of the chick embryo in a different manner from FGF1, -2, and -4 and activates the expression of Fgf8, an AER marker gene, in a cultured flank ectoderm without the mesoderm. Remarkably, FGF7 and FGF10 applied in the back induced an ectopic AER in the dorsal median ectoderm. Our results suggest that FGF7 and FGF10 directly induce the AER in the ectoderm both of the flank and of the dorsal midline and that these two regions have the competence for AER induction. Formation of the AER of the dorsal median ectoderm in the chick embryo is likely to appear as a vestige of the dorsal fin of the ancestors.

Animals↗

PubMed1999

Small GTPase RhoD suppresses cell migration and cytokinesis.

Rho family small GTPases regulate organization of the actin cytoskeleton. Among them, RhoA plays essential roles in the formation of the actin stress fibers, the associated focal adhesions, and the contractile rings necessary for cytokinesis. Recently, RhoD, a novel member of Rho family has been identified. The amino acid sequences of its effector domain is distinct from those of the other Rho family proteins, suggesting its unique cellular functions. Introduction of the constitutively active form of RhoD(G26V) into fibroblasts by microinjection or transfection resulted in disassembly of the actin stress fibers and the focal adhesions, whereas the dominant negative form of RhoD(T31K) did not affect these structures. The degree of cell migration assessed by the phagokinetic tracks on a substrate covered with gold particles was diminished by the expression of RhoD(G26V) but not by RhoD(T31K). Thus, cytoskeletal alterations including the loss of stress fibers and focal adhesions by RhoD seems to lead to the retardation of cell migration. Transfection of RhoD(G26V) cDNA into cultured cells also induced multinucleation. Moreover, RhoD(G26V) microinjected into fertilized eggs and embryos of Xenopus laevis caused cleavage arrest only in the injected cells, and the uncleaved cells contained multiple nuclei. These results imply that RhoD does not affect nuclear division but can interfere with cytokinesis presumably by preventing the formation of the actin-based contractile ring. Enhancement of the stress fibers by RhoA or RhoA-activating lysophosphatidic acid was reversed by the transfection of RhoD cDNA. Accordingly, the cellular functions of RhoD are likely to be antagonistic to those of RhoA.

Amino Acid Sequence↗

PubMed1999

Large-scale production of N-acetyllactosamine through bacterial coupling.

A large-scale production system of N-acetyllactosamine, a core structure of various oligosaccharides, was established by a whole-cell reaction through the combination of recombinant Escherichia coli strains and Corynebacterium ammoniagenes. Two recombinant E. coli strains over-expressed the UDP-Gal biosynthetic genes and the beta-(1-->4)-galactosyltransferase gene of Neisseria gonorrhoeae, respectively. C. ammoniagenes contributed the production of UTP from orotic acid. N-Acetyllactosamine was accumulated at 279 mM (107 g L-1) after a 38 h reaction (2.5 L in volume) starting from orotic acid, D-galactose, and 2-acetamido-2-deoxy-D-glucose.

Acetylglucosamine↗

PubMed1999

Uncoupling of transfer of the presequence and unfolding of the mature domain in precursor translocation across the mitochondrial outer membrane.

Translocation of mitochondrial precursor proteins across the mitochondrial outer membrane is facilitated by the translocase of the outer membrane (TOM) complex. By using site-specific photocrosslinking, we have mapped interactions between TOM proteins and a mitochondrial precursor protein arrested at two distinct stages, stage A (accumulated at 0 degrees C) and stage B (accumulated at 30 degrees C), in the translocation across the outer membrane at high resolution not achieved previously. Although the stage A and stage B intermediates were assigned previously to the forms bound to the cis site and the trans site of the TOM complex, respectively, the results of crosslinking indicate that the presequence of the intermediates at both stage A and stage B is already on the trans side of the outer membrane. The mature domain is unfolded and bound to Tom40 at stage B whereas it remains folded at stage A. After dissociation from the TOM complex, translocation of the stage B intermediate, but not of the stage A intermediate, across the inner membrane was promoted by the intermembrane-space domain of Tom22. We propose a new model for protein translocation across the outer membrane, where translocation of the presequence and unfolding of the mature domain are not necessarily coupled.

Amino Acid Sequence↗

PubMed1999

Differential expression of sialoglycoproteins in the rat hippocampus and its changes during aging.

The distribution pattern of sialylated glycoproteins in the hippocampal formation of 9-week-old and 29-month-old rats was examined using Maackia amurensis lectin, specific for Sia alpha2-3Gal, and Sambucus sieboldiana agglutinin specific for Sia alpha2-6Gal(GalNAc), and a monoclonal antibody for a linear homopolymer of alpha2-8-linked sialic acid. Each lectin and antibody showed different and characteristic staining patterns. On comparison of younger and older animals, different staining patterns were revealed. These results indicate that sialoglycoproteins with different sialyl linkages were expressed in distinct regions of the rat brain and their expression patterns changed with aging.

Aging↗

PubMed1999

Two distinct mechanisms operate in the reactivation of heat-denatured proteins by the mitochondrial Hsp70/Mdj1p/Yge1p chaperone system.

The yeast mitochondrial Hsp70, Ssc1p, functions as a molecular chaperone with its partner proteins, Mdj1p (DnaJ homologue) and Yge1p (GrpE homologue). We have purified a mature form of Ssc1p from yeast mitochondria and those of Mdj1p and Yge1p from Escherichia coli overexpresser cells. With these purified components of the mitochondrial Hsp70 chaperone system, we have succeeded in reconstituting their chaperone functions in the protection of firefly luciferase against thermal damage in vitro. Heat-denatured luciferase is prevented from irreversible aggregation and is maintained in a refolding-competent state by Ssc1p and/or Mdj1p at 42 degreesC. Luciferase denatured at 42 degreesC is actively reactivated by Ssc1p, Mdj1p and/or Yge1p after lowering the temperature to 25 degreesC. The reactivation process of heat-denatured luciferase shows two-phase kinetics. The slow refolding process requires either Ssc1p or Mdj1p at 42 degreesC but the presence of Ssc1p, Mdj1p and Yge1p, and ATP hydrolysis, is essential at 25 degreesC. The slow refolding of luciferase involves multiple rounds of formation and dissociation of the complex between luciferase and Mdj1p/Ssc1p. On the other hand, the fast refolding process is most enhanced when luciferase is incubated with Ssc1p alone at 42 degreesC, and it requires neither the assistance of Mdj1p and Yge1p nor ATP hydrolysis. We have observed a similar two-pathway reactivation of heat-denatured luciferase by the bacterial Hsp70 and the yeast cytosolic Hsp70 systems.

Adenosine Triphosphate↗

PubMed1999

Detection of novel carbohydrate binding activity of interleukin-1.

Tamm-Horsfall glycoprotein (THGP) and the oligosaccharide fraction liberated from THGP by hydrazinolysis inhibited tetanus toxoid-induced T cell proliferation. Intact THGP showed approximately 100-fold more inhibitory activity than the free oligosaccharides. After fractionating the oligosaccharides by anion-exchange column chromatography, the inhibitory activity could be detected in a sialidase-resistant acidic oligosaccharide fraction (fraction AR). The inhibitory activity of fraction AR was not observed when the fraction was added to the T cell culture medium 24 h after the addition of tetanus toxoid. Increased concentration of interleukin (IL) 1beta and decreased concentration of IL-2 were observed in the T cell culture medium after the addition of fraction AR. The oligosaccharides in fraction AR also inhibited the growth of an IL-1-dependent cell line, D10-G4. These results strongly suggested that the oligosaccharides in fraction AR bind to IL-1beta and suppress its cytokine activity. IL-1beta actually bound to the fraction AR immobilized on an amino-bonded thin layer plate. Fractionation of the oligosaccharides indicated that only oligosaccharides containing an N-acetylgalactosamine residue and a sulfate residue bound specifically to IL-1beta. Removal of either the sulfate residue or the N-acetylgalactosamine residue from the oligosaccharides abolished both the proliferation-inhibition and IL-1beta binding activities. Since IL-1beta did not bind to thyroid-stimulating hormone, which has the sulfate group at C-4 of the N-acetylgalactosamine residue in its N-linked sugar chains, the binding of IL-1beta toward oligosaccharides in fraction AR was considered to be highly specific.

Acetylation↗

PubMed1999

CBP-dependent and independent enhancing activity of steroid receptor coactivator-1 in thyroid hormone receptor-mediated transactivation.

Full-length of steroid receptor coactivator-1 (F-SRC-1) has been shown to interact with thyroid hormone receptors (TRs) in a ligand-dependent manner and to stimulate receptor-dependent transcription. To identify functional domains of F-SRC-1, several internal deletion mutants of F-SRC-1 were constructed. Although in vitro pull down assay with TR showed interaction of all of these mutants with TR, lack of mid legion (amino acids 398-1172) lost enhancing activity of TR-mediated transcription in a transient transfection assay. However, F-SRC-1 mutant lacking CBP-interacting domain still preserved enhancing activity. Surprisingly, F-SRC-1 mutants also increased basal level of viral promoter activity depending upon their deleted region. Yeast activation function assay revealed that these F-SRC-1 mutants had intrinsic activation function when bound to DNA. Analyses of small fragments of F-SRC-1 identified three separable activation domains. In vitro binding assay showed that TBP and TFIIB bound to C-terminal half of F-SRC-1. These results suggest that F-SRC-1 can function via both CBP-dependent and independent manners using various sets of activation domains and that direct interactions between F-SRC-1 and TBP or TFIIB may not be important for CBP-independent transcription.

Animals↗

PubMed1999

Expression of MUC2 gene in gastric regenerative, metaplastic, and neoplastic epithelia.

It has been reported that MUC2 mucin is expressed in goblet cells of gastric intestinal metaplasia, but not in its normal epithelium. To confirm this finding, we have examined the expression of the MUC2 gene by reverse transcriptase-polymerase chain reaction and immunohistochemical methods in gastric tissues obtained by routine upper gastrointestinal tract endoscopy and compared the results with pathological findings based on hematoxylin and eosin (H&E) staining. In 16.7% of the tissue specimens tested, MUC2 mRNA was detected in spite of the absence of intestinal metaplasia in HE specimens. A possible explanation for this was the identification by immunohistochemistry of MUC2 protein in regenerative gastric mucosal cells in biopsies that did not contain intestinal metaplasia. Sialyl-Le(x) epitope, which is suggested to be located on MUC2 mucin core protein (MUC2 protein), was also immunohistochemically detected in both goblet cells of intestinal metaplasia and regenerative epithelium. With regard to carcinoma, MUC2 protein was predominantly expressed in intestinal-type adenocarcinoma. These data indicate that MUC2 mucin is expressed in gastric regenerative epithelium in addition to intestinal metaplasia and intestinal type adenocarcinoma.

Adult↗

PubMed1999

Comparison of suppressive effects of a new anti-inflammatory compound, FR167653, on production of PGE2 and inflammatory cytokines, human monocytes, and alveolar macrophages in response to endotoxin.

FR167653 [1-[7-(4-fluorophenyl)-1,2,3,4-tetrahydro-8 (4-pyridyl) pyrazoro [5-1-c] [1,2,4] triazin-2-yl]-2-phenylethanedion sulfate monohydrate] was developed to inhibit proinflammatory cytokine production. However, the effects of FR167653 on prostanoid production are unclear. We investigated the effect of FR167653 on proinflammatory cytokine and prostaglandin (PG) production by lipopolysaccharide (LPS)-stimulated human peripheral blood monocytes and alveolar macrophages (AM) from the same individuals, and compared the effects in monocytes and AM. FR167653 inhibited interleukin-1beta and tumor necrosis factor alpha production. The effect on PGE2 production was assessed by four parameters. FR167653 inhibited PGE2 synthesis and LPS induction of cyclooxygenase activity. Western and Northern blot analyses revealed that LPS induction of cyclooxygenase-2 expression was attenuated by this compound. The suppression in monocytes was greater than that in AM. We concluded that the reduction of LPS-induced PGE2 synthesis by FR167653 was due to inhibition of cyclooxygenase-2 production. These results show that FR167653 may be therapeutically useful for inhibiting production of both inflammatory cytokines and PG production in inflammatory diseases.

Anti-Inflammatory Agents, Non-Steroidal↗

PubMed1999

Clustering of Acanthamoeba isolates from human eye infections by means of mitochondrial DNA digestion patterns.

A total of 90 Acanthamoeba isolates from human eye infections from 15 countries were clustered into distinct genotypes according to their mtDNA restriction fragment patterns. Closely related digestion phenotypes (sequence difference 0.1-1.5%) were integrated into a single genotype, whereas phenotypes with greater than 4.76% difference were considered distinct. Approximately 80% of the human isolates studied fell into 7 of 22 genotypes, indicating that virulence may be associated with specific clusters of cladistic groups of Acanthamoeba. This technique is useful for large-scale surveying of this particular pathogen.

Acanthamoeba↗

PubMed1999

Effects of a mixture of organisms, Lactobacillus acidophilus or Streptococcus faecalis on delta6-desaturase activity in the livers of rats fed a fat- and cholesterol-enriched diet.

The effect of a mixture of organisms (a probiotic mixture) comprising Bacillus, Lactobacillus, Streptococcus, Clostridium, Saccharomyces, and Candida (10(7-8) colony-forming units/g rice bran of each component) on delta6-desaturase activity in liver microsomes was compared with those of Lactobacillus acidophilus and Streptococcus faecalis. There were four treatment groups. Each group of these rats received rice bran (control), the mixture of organisms, L. acidophilus, or S. faecalis (30 g/kg) along with a fat- and cholesterol-enriched diet for 4 wk. The serum total cholesterol concentration of the group fed the mixture of organisms was reduced by 15-33% compared with the other groups at the end of the 4-wk feeding period (P<0.05). The proportion of palmitic acid in the serum phosphatidylcholine (PC) for the control group was significantly higher than those of the other groups. The proportion of arachidonic acid in the serum PC for the mixed-organism group was also significantly higher than those of the other groups. The proportion of arachidonic acid in the liver PC for the mixed-organism group was significantly higher than those of the control and S. faecalis groups. The ratio of arachidonic acid/linoleic acid was significantly higher in the liver PC of rats fed the mixed organisms compared with the control group (P<0.05). The delta6-desaturase activity in the liver microsomal fraction of the mixed-organism group was significantly higher than those of the other groups. The delta6-desaturase activity correlated positively with the ratio of arachidonic acid/linoleic acid of liver PC, the correlation coefficient (r) being 0.819 (P<0.001). The results indicate that the effect of the mixture of organisms was to increase delta6-desaturase activity and serum arachidonic acid and decrease cholesterol compared to the other organisms and control, but the mechanism whereby the enzyme activity was related to serum cholesterol does not appear to have been explored.

Animals↗

PubMed1999

Sleeping environments as risk factors of sudden infant death syndrome in Japan.

For the purpose of assessing the risk factors of the sleeping environment in SIDS, we performed a population-based, case-control study. Index cases comprised 56 SIDS cases which were diagnosed on the basis of autopsies during a period of 11 years at the Department of Legal Medicine, Faculty of Medicine, University of the Ryukyus. Control cases comprised infants who were examined at regional health centers for Infant Health Screening. The incidence of SIDS obtained was 0.25 per 1000 live births, which was comparable to that obtained in other districts having the established medical examiner's system. The proportion of prone sleeping was 81.0% and 38.3% in the SIDS and control groups, respectively, and an odds ratio was calculated as 10.4 (99% confidence interval, 3.9 to 37.6). This indicates that prone sleeping is a risk factor of SIDS, in Japan, as reported in various countries. Various kinds of bedclothes were used in this country, and our survey of bedclothes in the control revealed their improper use for infants to sleep, particularly with the prone position. In addition, the co-sleeping habit, which was not uncommon in Japan, seems to contribute to certain deaths of infants whose causes of death were controversial. In the investigation of SIDS, therefore, the sleeping environments, such as bedclothes and the co-sleeping habit, as well as the sleeping position should be taken into consideration as risk factors.

Journal Article↗