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Biomedical subjects

Y Niwa

Publications and source records attributed to Y Niwa.

At least 109 records · Page 6Linked to original sources

The effect of nilvadipine, a calcium-channel blocker, on the hemodynamics of retrobulbar vessels in normal-tension glaucoma.

PURPOSE: To investigate the effect of nilvadipine, a calcium-channel blocker, on the hemodynamics of retrobulbar vessels in normal-tension glaucoma. METHODS: Twenty-five patients who prospectively met the enrollment criteria underwent color Doppler imaging of the retrobulbar vessels before and after receiving 4 weeks of treatment with 2 mg oral nilvadipine twice daily. RESULTS: Nilvadipine significantly increased the end-diastolic velocity in the central retinal artery and a short posterior ciliary artery. It significantly reduced the resistance index in the central retinal artery and posterior ciliary arteries, but not in the ophthalmic artery. The calculated ocular perfusion pressure was not affected. CONCLUSION: Oral nilvadipine reduces vascular resistance in distal retrobulbar arteries in normal-tension glaucoma without affecting more proximal blood vessels. Therefore, nilvadipine may have a beneficial effect on the hemodynamics of retrobulbar vessels in normal-tension glaucoma.

Administration, Oral↗

[Terminal home care "Ikigai no Yoake--The real aim of our life"--case report].

It is difficult to provide home care especially for terminal cancer patients as their physical conditions deteriorate due to the cancer. It is important to enhance the will of home care providers to make this possible. Although a universal method has been worked out, personal and religious beliefs have made it difficult to create an effective method. We would like to introduce our experience in the paper entitled "Ikigai no Yoake" and subtitled "Influence of scientific study of reincarnation on view of life" by Fumihiko Ida of Fukushima University. It has encouraged home care providers and made it possible to provide home care to terminal patients along with benefits.

Advance Directives↗

[Benefits and problems of "home care score"--discussion of deviation from home care].

Care ability is the most important factor in providing home care. This ability to care can be classified into family care and community care. The "Home care score" is a system developed through the comprehensive scientific study on long life by the Ministry of Health and Welfare in order to evaluate such care ability. We have scored 71 patients since August 1997 until February 1998, and we found benefits and problems with the system. Here are the benefits: (1) General evaluation can be made on status of patients and home care providers. (2) Comparison of large numbers of patients can be easily made. (3) Helpful to work out practical support. On the other hand, the system has the following problems: (1) Scoring can differ by subjective evaluation. (2) Difficulty to reflect status of disease deterioration and so on. The factors of deviation from home care are: (1) of disease. (2) Loss of desire to provide care. We believe it is important for the home care nurse to intervene so that the will to provide care will not weaken until the end.

Aged↗

Leaf-specifically expressed genes for polypeptides destined for chloroplasts with domains of sigma70 factors of bacterial RNA polymerases in Arabidopsis thaliana.

Genes for sigma-like factors of bacterial-type RNA polymerase have not been characterized from any multicellular eukaryotes, although they probably play a crucial role in the expression of plastid photosynthesis genes. We have cloned three distinct cDNAs, designated SIG1, SIG2, and SIG3, for polypeptides possessing amino acid sequences for domains conserved in sigma70 factors of bacterial RNA polymerases from the higher plant Arabidopsis thaliana. Each gene is present as one copy per haploid genome without any additional sequences hybridized in the genome. Transient expression assays using green fluorescent protein demonstrated that N-terminal regions of the SIG2 and SIG3 ORFs could function as transit peptides for import into chloroplasts. Transcripts for all three SIG genes were detected in leaves but not in roots, and were induced in leaves of dark-adapted plants in rapid response to light illumination. Together with results of our previous analysis of tissue-specific regulation of transcription of plastid photosynthesis genes, these results indicate that expressed levels of the genes may influence transcription by regulating RNA polymerase activity in a green tissue-specific manner.

Amino Acid Sequence↗

Intrinsic nucleoside diphosphate kinase-like activity as a novel function of 14-3-3 proteins.

14-3-3 proteins play a role in many cellular functions as molecular chaperone and adapter proteins: they bind to and modulate several proteins involved in cell proliferation and differentiation, and also function ATP-dependently in targeting of precursors to mitochondria. We show here that 14-3-3 purified from a human lymphoblastoma and also its recombinant tau isoform exhibited intrinsic nucleoside diphosphate (NDP) kinase-like activity. 14-3-3 proteins preferentially catalyzed the transfer of the gamma-phosphate group from ATP, dATP or dGTP to all nucleoside diphosphates and this transfer involved acid-labile phosphoenzyme intermediates. They also simultaneously catalyzed the reverse reaction of ATP hydrolysis. These properties of 14-3-3 are similar to those of NDP kinase, but not to those of adenylate kinase.

14-3-3 Proteins↗

Chromosomal mapping of genes in the RBCS family in Arabidopsis thaliana.

In Arabidopsis thaliana, four genes have been identified in the RBCS gene family, one being assigned to subfamily RBCS-A and the other three to subfamily RBCS-B (1B, 2B and 3B). To determine the chromosomal location of these genes, hybridization analysis with CIC YAC high-density filters was carried out for the RBCS-A gene, and CAPS analysis for the three RBCS-B genes, based on the finding that restriction fragment length polymorphism is present in the upstream region of the gene RBCS-3B. The RBCS-A gene was mapped at 100.8 cM from the top of chromosome 1 and the three RBCS-B genes at 62.70 cM from the top of chromosome 5.

Arabidopsis↗

Regression of superficial gastric MALT lymphoma with unsuccessful eradication therapy for Helicobacter pylori infection.

A 51-year-old man had a reddish flat granular lesion in the stomach on endoscopic examination. Histology of biopsied specimen confirmed the diagnosis of low-grade B-cell gastric lymphoma of mucosa-associated lymphoid tissue (gastric MALT lymphoma) and simultaneous infection with Helicobacter pylori. He was given antibiotic treatment. Five weeks later, endoscopy and histology of biopsied specimen showed eradication of H. pylori, and the tumor had regressed. Six months later, H. pylori reemerged, but the tumor had not recurred. After the second antibiotic therapy, H. pylori has been eradicated. The lymphoma has been in remission for 14 months.

Anti-Bacterial Agents↗

Endoscopic ultrasonography in the diagnosis of submucosal lesions of the large intestine.

BACKGROUND: Although reports of colorectal submucosal tumors have increased since the development of endoscopic examinations, precise diagnosis of these lesions remains difficult. In this study, we evaluated the usefulness of endoscopic ultrasonography (EUS) in the diagnosis of submucosal lesions of the large intestine. METHODS: From September 1989 to June 1996, EUS was performed in 46 patients who were suspected to have submucosal lesions by barium enema or colonoscopy. Twenty-seven of the 46 cases were confirmed histologically by endoscopic or surgical resection, and their ultrasonographic images were compared with resection specimens. RESULTS: Lipomas (n = 15) were visualized as hyperechoic masses and lymphangiomas (n = 9) visualized as cystic lesions with septal structures as characteristic findings. The EUS images of leiomyomas (n = 6), leiomyosarcomas (n = 3), and enteric endometriosis (n = 7) were all hypoechoic masses in the fourth layer. Leiomyosarcomas tended to be larger and more inhomogeneous than leiomyomas. Enteric endometriosis was shaped like a spindle or a half-moon, and myogenic tumors were lobulated when the lesions were large. Recurrences of colorectal carcinoma (n = 3), malignant lymphomas (n = 2), and an appendiceal mucocele (n = 1) were examined. CONCLUSIONS: EUS is useful in the diagnosis of submucosal lesions of the large intestine because it provides precise information about these lesions.

Adult↗

Inhibition of tryptase TL2 from human T4+ lymphocytes and inhibition of HIV-1 replication in H9 cells by recombinant aprotinin and bikunin homologues.

The serine esterase TL2 from human T4+ lymphocytes is a binding component to HIV-1 glycoprotein gp120 and seems to play a role in the HIV-1 infection mechanism. Recombinant variants of the Kunitz-type serine proteinase inhibitor aprotinin were investigated for their ability to inhibit tryptase TL2 and the binding of gp120 to this enzyme. Furthermore, the viral replication of HIV-1 was investigated H9 cell cultures under the influence of recombinant aprotinin and bikunin variants. In contrast to native aprotinin, the recombinant variant [Arg15, Phe17, Glu52] aprotinin with a reactive-site sequence homologous to the V3 loop of HIV-1 gp120 showed a specific inhibition of tryptase TL2 (> 80%). However, the [Leu15, Phe17, Glu52] aprotinin variant with hydrophobic subsites was the most potent inhibitor of the binding of gp120 to tryptase TL2 (68%). Our results show that the enzyme activity of purified tryptase TL2 is inhibited not only by variants with basic amino acids, but also those with hydrophobic residues in the reactive-site region. Therefore, tryptase TL2 is not a typical trypsin-like or chymotrypsin-like protease. Investigations on inhibition of HIV-1 replication in H9 cell cultures showed that tryptase TL2 is involved in the mechanism of virus internalization into human lymphocytes. The [Leu15, Phe17, Glu52] aprotinin showed a significant retardation of syncytium formation over a period of 5 days in a 1 micro M concentration. Similar investigations were performed with recombinant variants of bikunin, the light chain of human inter-alpha-trypsin inhibitor. Only the single-headed variant [Arg94] delta 2 bikunin inhibited slightly the syncytium formation over a period of 2 days in a 2.2 micro M concentration. Wild-type bikunin and all full-length variants showed no effect, possibly due to steric hindrance by the second domain of the double-headed inhibitor.

Amino Acid Sequence↗

Evidence for transcriptional regulation of plastid photosynthesis genes in Arabidopsis thaliana roots.

Mechanisms underlying suppressed levels of transcripts for plastid photosynthesis genes in nongreen tissues such as roots and calli were analyzed in Arabidopsis thaliana, a plant suitable for further genetic dissection. A region encoding promoters of rbcL, the gene encoding the large subunit of ribulose-1,5-biphosphate carboxylase/oxygenase, and the atpB/E operon for the beta and epsilon subunits of coupling factor one were cloned and sequenced. Transcripts for rbcL, atpB/E, and psbA, the gene for the D1 protein in the photosystem II reaction center, were barely detectable in roots of A. thaliana, whereas 16S rRNA was detected at a low level. The run-on transcription experiment revealed that expression of rbcL, atpB/E, and psbA was regulated at transcription. The copy number of plastid DNA in roots was one-fifth that in green leaves on the basis of total cellular DNA, suggesting that in the latter the DNA copy-number regulation also exists in plastid gene expression. Digestion of DNA with methyl-sensitive and -insensitive isoschizomeric endonucleases and subsequent polymerase chain reaction, as well as in vitro transcription of plastid DNAs with Escherichia coli RNA polymerase, resulted in no evidence of regulation by DNA modification. In spite of predominant suppression of expression of rbcL, atpB/E, and psbA at transcription in roots and calli, 16S rRNA levels were decreased because of low RNA stability.

Arabidopsis↗

Inhibitory effect of local anaesthetics on reactive oxygen species production by human neutrophils.

BACKGROUND: Reactive oxygen species (ROS) generated from neutrophils accumulated in various major organs are thought to play a pivotal role in the pathogenesis of host auto-injury. Lidocaine has been shown to reduce the injury. We investigated the effect of local anaesthetics (lidocaine, mepivacaine and bupivacaine) on ROS production by neutrophils using an in vitro system. METHODS: We measured the production of superoxide (ferricytochrome c method), hydrogen peroxide (H2O2: scopoletin fluorescence technique), and hydroxyl radical (OH.: ethylene gas method) by neutrophils isolated from human adult volunteers in the absence and presence of lidocaine (2-200 micrograms/mL), mepivacaine (3-300 micrograms/mL), and bupivacaine (3-300 micrograms/mL). We also measured the ROS generation in a cell-free (xanthine-xanthine oxidase) system. RESULTS: Lidocaine and mepivacaine at higher levels significantly decreased the production of ROS by neutrophils. However, these local anaesthetics at clinically relevant blood concentrations had no effect on the levels of ROS. Furthermore, lidocaine and mepivacaine failed to reduce ROS generated by the cell-free system. Bupivacaine did not decrease ROS generation by either generating system. CONCLUSION: In conclusion, in the present in vitro system, only concentrations of lidocaine and mepivacaine 100-fold higher than clinically feasible ones reduced ROS production by human neutrophils. However, the local anaesthetics at clinically relevant blood concentrations had no suppressive effect. Further studies using in vivo systems are required to elucidate the inhibitory effects of local anaesthetics on ROS generation in clinical settings.

Adult↗

Relationship between local immune response to Helicobacter pylori and the diversity of disease: investigation of H. pylori-specific IgA in gastric juice.

In order to evaluate the relationship between local immune response to Helicobacter pylori and the diversity of disease, 77 asymptomatic subjects who underwent a health examination were studied. Helicobacter pylori-specific IgG in serum and H. pylori-specific IgA in gastric juice were measured by ELISA, and the measured IgA titre was classified into two grades, low or high. Histological classification of gastritis was performed according to the Sydney system. Cytokines in gastric juice were also measured, and the cytotoxin-associated gene A (cagA) status of H. pylori was tested by PCR. Of the 65 subjects who were positive for H. pylori-specific IgG in serum, 38 (58.5%) were classified as H. pylori-specific IgA low titre in gastric juice and 27 (41.5%) had high titres. In the IgG-positive, IgA-low group, the rate of peptic ulcers (especially duodenal ulcers) in endoscopic findings was higher (P < 0.05); the score of activity and the density of H. pylori were higher (P < 0.001 and P < 0.05, respectively); the score of metaplasia was lower (P < 0.05); and the level of interleukin-1 beta was lower (P < 0.05) than in the IgG-positive, IgA-high group. The positive rate of the cagA gene was 84.4% and there was no significant difference between the two groups. There were differences in endoscopic and histological findings between the IgG-positive, IgA-low and the IgG-positive, IgA-high groups. It is suggested that persons infected with H. pylori can be divided into two different states of disease according to local immune response.

Antibodies, Bacterial↗

Kinetics of fibroblasts in ulcer healing in rats: interference with indomethacin.

OBJECTIVE: The authors reported that the repair of submucosal tissue is important in ulcer healing. The kinetics of fibroblasts have not been well known in gastric ulcer healing. The effect of subcutaneous administration of indomethacin (1 mg/kg) to submucosal tissue was also examined. METHOD: Immunohistological staining was done using the antibodies for the proliferating cell nuclear antigen prolyl 4-hydroxylase and alpha-smooth muscle actin in acetic acid induced gastric ulcers in rats. The terminal deoxynucleotidyl transferase mediated deoxyuridine triphosphate-biotin nick end labeling method was also employed. RESULTS: The granulation tissue consisted predominantly of lymphocytes in indomethacin-treated rats. In the control group, proliferative cell nuclear antigen positive cells were at their peak 5 days after ulcer formation (day 5). Prolyl 4-hydroxylase positive fibroblasts were maximal on day 10. Alpha-smooth muscle actin positive myofibroblasts were few on day 5 and increased on day 15. In terminal deoxynucleotidyl transferase mediated deoxyuridine triphosphate-biotin nick end labeling, positive cells were sporadically seen on days 20 and 30. In indomethacin-administered rats, the fibroblasts proliferated weakly. The numbers of prolyl 4-hydroxylase and alpha-smooth muscle actin positive spindle-shaped cells were fewer and their appearance delayed. Nick end labeling positive cells were few before day 30 and sporadically observed on days 50 and 80. CONCLUSION: The results indicate that fibroblasts differentiated into several phenotypes and finally underwent apoptosis.

Acetic Acid↗

Three-dimensional endoscopic ultrasonography of gastrointestinal lesions using an ultrasound probe.

BACKGROUND: This study was carried out to ascertain the clinical usefulness and problems of three-dimensional (3D) images obtained with the ultrasound probe. METHODS: Eighteen resected specimens and 21 patients were examined. After the images were recorded on video, 3D displays were produced using a workstation. RESULTS: In the in vitro study the surface images were quite consistent with the macroscopic findings in 17 cases. In 2 esophageal cancers, 7 of 10 gastric cancers, and 2 colonic cancers the depth of tumor invasion was assessed accurately from the reconstructed images. In the in vivo study although 3 D display had some limitations, it was useful in esophageal and rectal lesions. CONCLUSIONS: This new diagnostic method could visualize the 3D extent of gastrointestinal lesions and appears to have a useful clinical application.

Adult↗

[Trigeminal neuralgia caused by tortuous vertebrobasilar system--the clinical and imaging features].

Ten (6.8%) out of 146 patients with trigeminal neuralgia (TN) who underwent SPGR-MRI and 3D-TOF-MRA from August 1993 to October 1996, were found to have vascular compression caused by a tortuous vertebrobasilar system (TVBS). They were mostly males, demonstrated left-sided predominance, and had ipsilateral hemifacial spasm, compared with other 52 patients whose offending arteries were either superior cerebellar artery (SCA), anterior inferior cerebellar artery (AICA)or posterior inferior cerebellar artery (PICA). The patients who showed vascular compression by TVBS, presented an artery which compresses and dislocates the rootentry zone (REZ) of the trigeminal nerve, presses the brain stem at REZ and simultaneously compresses the REZ of the facial nerve. In addition, the diameters of the two branches of vertebrobasilar artery were not equal. These features indicate that the atherosclerotic change of the offending artery in TN caused by TVBS is more severe than that caused by SCA, AICA or PICA. This change causes an irregular running of artery which leads a strong compression of the trigeminal nerve REZ and of the brain stem. Consequently, the facial nerve REZ is severely affected leading to the presence of tic convulsif in TN caused by TVBS.

Aged↗

T-cell membrane-associated serine protease, tryptase TL2, binds human immunodeficiency virus type 1 gp120 and cleaves the third-variable-domain loop of gp120. Neutralizing antibodies of human immunodeficiency virus type 1 inhibit cleavage of gp120.

It has been suggested that the third variable domain (V3) loop of human immunodeficiency virus type 1 (HIV-1) gp120 has to interact with a cell-surface-associated protease(s) that acts as a cofactor after binding of gp120 to the CD4 receptor during entry of HIV-1 into susceptible cells. We isolated the membrane-associated serine protease, tryptase TL2, from human CD4-positive lymphocytes. This enzyme specifically binds gp120 through interaction with its V3 domain. To investigate the role of tryptase TL2 in HIV infection, we examined the affinity of the interaction and the proteolytic susceptibility of various recombinant gp120 expressed in mammalian cells to the enzyme, and we determined the cleavage sites. Tryptase TL2 bound gp120 with an apparent dissociation constant of 38 nM. The affinity was lower than that of gp120 for CD4 which suggests that gp120 initially binds to CD4, followed by interaction with tryptase TL2 which is localized close to CD4 on the cell surface. After binding, tryptase TL2 cleaved recombinant gp120 expressed in mammalian cells into two protein species of 70 kDa and 50 kDa but did not cleave gp120 expressed in insect cells, which indicates that the structure of the oligosaccharides linked to the polypeptide backbone of gp120 affects the proteolytic susceptibility. Cleavage was specifically inhibited by a neutralizing antibody against the V3 loop. Cleavage-site determination revealed that tryptase TL2 cleaved gp120 at various sites in the V3 in a strain-dependent manner. The amino acid variability at the cleavage site(s) in almost all HIV-1 isolates was restricted to amino acids which are susceptible to the chymotryptic and/or tryptic activities of tryptase TL2.

Amino Acid Sequence↗

Engineered GFP as a vital reporter in plants.

BACKGROUND: The green-fluorescent protein (GFP) of the jellyfish Aequorea victoria has recently been used as a universal reporter in a broad range of heterologous living cells and organisms. Although successful in some plant transient expression assays based on strong promoters or high copy number viral vectors, further improvement of expression efficiency and fluorescent intensity are required for GFP to be useful as a marker in intact plants. Here, we report that an extensively modified GFP is a versatile and sensitive reporter in a variety of living plant cells and in transgenic plants. RESULTS: We show that a re-engineered GFP gene sequence, with the favored codons of highly expressed human proteins, gives 20-fold higher GFP expression in maize leaf cells than the original jellyfish GFP sequence. When combined with a mutation in the chromophore, the replacement of the serine at position 65 with a threonine, the new GFP sequence gives more than 100-fold brighter fluorescent signals upon excitation with 490 nm (blue) light, and swifter chromophore formation. We also show that this modified GFP has a broad use in various transient expression systems, and allows the easy detection of weak promoter activity, visualization of protein targeting into the nucleus and various plastids, and analysis of signal transduction pathways in living single cells and in transgenic plants. CONCLUSIONS: The modified GFP is a simple and economical new tool for the direct visualization of promoter activities with a broad range of strength and cell specificity. It can be used to measure dynamic responses of signal transduction pathways, transfection efficiency, and subcellular localization of chimeric proteins, and should be suitable for many other applications in genetically modified living cells and tissues of higher plants. The data also suggest that the codon usage effect might be universal, allowing the design of recombinant proteins with high expression efficiency in evolutionarily distant species such as humans and maize.

Allium↗