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Biomedical subjects

Y Yamauchi

Publications and source records attributed to Y Yamauchi.

At least 37 records · Page 2Linked to original sources

Resazurin as an electron acceptor in glucose oxidase-catalyzed oxidation of glucose.

The behavior of resazurin (1) as an electron acceptor in glucose oxidase (GOD)-catalyzed oxidation of glucose under anaerobic conditions is described. When a mixture of 1, glucose, and GOD in phosphate buffer (pH 7.4, 0.1 M) was incubated at 25 degrees C, the resulting solution turned purple to fluorescent pink due to the deoxygenated product, resorufin (2). On incubation of 1 with GOD alone or with H2O2 under essentially the same conditions, no color change was seen, indicating that generation of 2 in the enzymatic reaction is brought about through reduction of 1 by the reduced form (GODred) of GOD, which was also supported by the voltammetric behavior of 1. However, it was found that the enzymatic transformation of 1 to 2 is of no practical use as an indicator reaction for glucose determination using only GOD due to a slow reaction of 1 with GODred. Based on a ping-pong type mechanism with a steady-state approximation, KM and kcat for 1 as an electron acceptor from GODred were estimated to be 15+/-1.3 microM and (5.0+/-0.5) x 10(-2) s(-1), respectively.

Electrons↗

Purification of an aminopeptidase preferentially releasing N-terminal alanine from cucumber leaves and its identification as a plant aminopeptidase N.

In this study, a highly active foliar aminopeptidase preferentially releasing N-terminal alanine from artificial substrates was purified and characterized from cucumber (Cucumis sativus L. suyo). The enzyme had a molecular mass of 200 kDa consisting of two subunits of 95 kDa. It was a metalloprotease the pH optimum of which was 8 to 9. It cleaved Ala-, Gly-, Met-, Ser-, Leu-, Lys-, and Arg artificial substrates. An internal amino acid sequence was similar to those of aminopeptidase N (clan MA, family M1) of microorganisms, and was very similar to that of a putative aminopeptidase N of Arabidopsis thaliana. From these results, the highly active aminopeptidase in cucumber leaves was identified to be a plant aminopepitdase N.

Alanine↗

An autopsy case of primary biliary cirrhosis with severe interstitial pneumonia.

A 74-year-old woman was admitted to our hospital because of interstitial pneumonia. She had a 14-year history of primary biliary cirrhosis (PBC) diagnosed histologically, with a positive test for anti-mitochondrial antibodies and elevated biliary enzyme activity. She also had a 7-year history of rheumatoid arthritis and a 26-year history of Sjögren's syndrome. Though the symptoms of these complications improved, the interstitial pneumonia deteriorated very quickly and the patient died of respiratory failure due to acute exacerbation of interstitial pneumonia when the activity of PBC decreased. We report this case because it is relatively rare for PBC to be complicated by severe interstitial pneumonia, and it may offer insight into the etiology of these diseases.

Aged↗

[Evaluation of canal wall down tympanoplasty with canal reconstruction for draining ear with middle ear cholesteatoma].

OBJECTIVES: We studied the postoperative stability of canal wall down tympanoplasty with canal reconstruction for middle ear cholesteatoma with preoperative otorrhea. SUBJECTS AND METHODS: 155 ears with middle ear cholesteatoma treated with canal wall down tympanoplasty with canal reconstruction were evaluated retrospectively. A comparison was made between the group of 80 ears which showed otorrhea, preoperatively, and the group of 75 without preoperative otorrhea. Problems observed in the tympanic membrane or reconstructed external auditory canal were evaluated both at the postoperative initial stage and more than 1 year after surgery. Postoperative hearing prognosis was also studied. RESULTS: 1) In the postoperative initial stage, local infection and necrosis of materials for canal reconstruction were significantly more likely to be observed in ears with preoperative otorrhea. 2) In ears with postoperative local infection, necrosis of materials for canal reconstruction occurred more frequently, and the period until drying of the reconstructed external auditory canal was significantly extended. 3) No significant difference was seen in postoperative status of the tympanic membrane and reconstructed ear canal at least 1 year after surgery. 4) The presence of preoperative otorrhea had no influence on hearing prognosis. CONCLUSIONS: When canal wall down tympanoplasty with canal reconstruction is used for ears with preoperative otorrhea, careful attention should be paid to local treatment at the postoperative initial stage. However, no significant problem occurred in the outcome of preoperative ear draining at least 1 year after surgery.

Adolescent↗

CJ-13,981 and CJ-13,982, new squalene synthase inhibitors.

Two new squalene synthase (SSase) inhibitors, CJ-13,981 (I) and CJ-13,982 (II), were isolated from the fermentation broth of an unidentified fungus CL 15036. They inhibited human liver microsomal SSase with IC50s of 2.8 and 1.1 microM, respectively, but showed no inhibitory activity against human brain protein farnesyltransferase (PFTase) at 100 microM. Based on FAB-MS and NMR analyses, the structures of I and II were determined to be 3-hydroxy-3,4-dicarboxy-15-hexadecenoic acid and 3-hydroxy-3,4-dicarboxyhexadecanoic acid, respectively.

Alkyl and Aryl Transferases↗

Identification and comparison of residues critical for cell-adhesion activities of two neutrophil CD66 antigens, CEACAM6 and CEACAM8.

CEACAM6 (CD66c) and CEACAM8 (CD66b) are cell-adhesion proteins on neutrophils that belong to the human carcinoembryonic antigen (CEA) family. CEACAM6 reveals homophilic adhesion and heterophilic adhesion to other CEACAM family antigens including CEACAM8, CEACAM1, and CEA, whereas CEACAM8 exhibits only heterophilic adhesion to CEACAM6. Here, we investigated and compared structural requirements for the homophilic adhesion of CEACAM6 and heterophilic adhesion between CEACAM6 and CEACAM8 at the amino acid level by using CHO transfectants expressing their mutant and chimeric proteins. The NH(2)-terminal domain (N-domain) of CEACAM6 expressed on a CHO cell was suggested to bind the N-domain of CEACAM6 or CEACAM8 on the opposing cell. By homologue-scanning mutagenesis, we found that the locations of the sequences critical for the adhesion of CEACAM6 to itself and to CEACAM8 are overlapped and that they are highly similar but not identical to the locations of the residues previously shown to be essential for the binding of CEACAM antigens to Opa proteins of pathogenic NEISSERIAE: Our findings imply that subtle differences in the N-domain sequences determine the specificity of the CEACAM antigens on neutrophils for interaction with the same or different CEACAM antigens and the bacterial proteins.

Amino Acid Sequence↗

[Comparison of five guidelines of community-acquired pneumonia].

The aim of this retrospective study was to elucidate the characteristics of five guidelines of community-acquired pneumonia: ATS (1993), ATS (2001), IDSA (1998), IDSA (2000) and the guidelines of the Japan Respiratory Society (2000). One hundred community-acquired pneumonia patients admitted to the International Medical Center of Japan were investigated in accordance with each set of guidelines based on the physical, laboratory, and chest radiography findings on the first day of treatment. According to the ATS (1993) guidelines, 33% of the cases were classified as "severe" pneumonia. On the other hand, according to the ATS (2001) guidelines, only 8% of the cases were classified as "severe" pneumonia. According to the IDSA guidelines, 35% of the patients were classified as "outpatients". Fluoroquinolone appears to be a very important antibiotic drug in the new guidelines of both ATS and IDSA. The scoring system of IDSA suggested a correlation between the patient's score and the pathogenic bacteria. According to the guidelines of the Japan Respiratory Society, 42% of the cases were classified as "severe" pneumonia. There are evident differences between these guidelines, and clinicians need to have a full understanding of their respective characteristics.

Community-Acquired Infections↗

Molecular cloning of ring finger protein 21 (RNF21)/interferon-responsive finger protein (ifp1), which possesses two RING-B box-coiled coil domains in tandem.

We have cloned the full length of a novel cDNA, named ring finger protein 21 (RNF21), composed of the RING finger-B box-coiled coil (RBCC) domain and the B30.2 domain, which are characteristic of the RBCC-B30.2 family. As a structural feature, the RNF21 cDNA possessed at least three kinds of isoforms, due to alternative splicing, consisting of the long form with the RBCC-RBCC-B30.2 domain, the medium form with the RBCC-B30.2 domain, and the short form with only the RBCC domain. Moreover, respective transcripts corresponding to the three isoforms were detected in various human organs by reverse transcription-PCR and Northern blot analyses. Interestingly, the medium form of the RNF21 mRNA expressed most predominantly was dramatically up-regulated within 8-16 h by interferon stimulation of HeLa cells. These findings suggest that RNF21 is a downstream gene that may mediate interferon's biological action.

Alternative Splicing↗

Molecular cloning of testis-abundant finger Protein/Ring finger protein 23 (RNF23), a novel RING-B box-coiled coil-B30.2 protein on the class I region of the human MHC.

We have identified a genomic DNA fragment, using the PCR method with degenerate oligonucleotide primers which contain the conserved sequence of the RING finger domain. Using the DNA fragment as a probe, a novel cDNA was cloned from human and mouse testis. The cDNA had a domain structure of the typical RING-B box-coiled coil (RBCC)-B30.2 domain and therefore was named testis-abundant finger protein (tfp). Indeed, the transcript was highly expressed in the testis, although it was also found ubiquitously in various organs by Northern blot analysis. The tfp gene was mapped at the class I region of the human MHC (major histocompatibility complex), within which some known RBCC-B30.2 proteins such as RFP, RFB30/HERF1, AFP, and HZF had been localized. These findings demonstrate that several RBCC-B30.2 proteins including tfp, which are non-HLA proteins, are clustered within the class I region of the human MHC.

Amino Acid Sequence↗

Metastable helium atom stimulated desorption of H+ Ion

H+ desorption induced by the impact of metastable helium atoms has been found for H(2)O/Na/Ni(110) coadsorption systems. The measurements were carried out using a time-of-flight technique and a pulsed-discharge type metastable helium atom ( He(*)) source. It is concluded that the H+ desorption by He(*) is induced by a hole created on the valence levels via the Auger deexcitation of He(*). The H+ desorption by He(*) may be understood within the framework of the Menzel-Gomer-Readhead model.

Journal Article↗

Identification of the novel developmentally regulated gene, Bdm2, which is highly expressed in fetal rat brain.

Most of the neurogenesis take place during the embryonic stage; the genes expressed predominantly in this stage may play important roles in the control of development of the central nervous system. Using a differential display method, we identified the novel rat gene, brain development-related molecule 2 (Bdm2), that is expressed more abundantly in the embryonic brain than in the adult brain. Full-length Bdm2 cDNA consists of 1842 base pairs (bp) and contains an open reading frame of 1260 bp. Northern blot analysis demonstrated that Bdm2 was strongly expressed in the late embryonic brain and was still detected at lower levels in an early postnatal period; in adults, Bdm2 mRNA was decreased to an undetectable level in brain, though the expression of this mRNA was revealed in other tissues. Level of Bdm2 mRNA was maintained during neuronal differentiation of mouse embryonal carcinoma cell P19, but decreased during the differentiation to glial and unidentified non-neuronal cells. In situ hybridization study demonstrated the wide distribution of Bdm2 mRNA in the embryonic brain; in the adult brain, the hybridization signals became more restricted to the hippocampus, olfactory bulb, cerebellum, and neocortex, almost coinciding with the regions where nascent and immature neurons are present. Thus, it appears likely that Bdm2 encodes a protein that is involved in both the regulation of growth of undifferentiated neural cells and the terminal differentiation of neuronal cells.

Amino Acid Sequence↗

cDNA and deduced amino acid sequences of apolipophorin-IIIs from Bombyx mori and Bombyx mandarina.

The cDNA sequence for apolipophorin-III from two strains of Bombyx mori (N4 and P50) and the Japanese and Chinese strains of Bombyx mandarina were determined. Both the cDNA and deduced amino acid sequences of the four apolipophorin-IIIs were highly similar (95-98%). The four Bombyx sequences also showed significant similarity to the sequence of apolipophorin-III from another lepidopteran, Manduca sexta (83-84%), particularly in the five amphipathic alpha-helices that are proposed to play a critical role in the binding of apolipophorin-III to lipophorin. In the coding region, the nucleotide sequences for the Chinese strain of B. mandarina and the P50 strain of B. mori were identical, supporting the suggestion that P50 is the current strain most closely related to the original domesticated strain. The N4 strain of B. mori is more closely related to these two strains than is the Japanese strain of B. mandarina, suggesting that Japanese strain of B. mandarina separated from the Chinese strain of B. mandarina before domestication of B. mori. Arch.

Amino Acid Sequence↗

Immunohistological study of intervertebral disc herniation of lumbar spine.

In order to observe histological changes in the extruded and sequestrated intervertebral disc, we conducted pathological and immunological examinations of herniated disc materials taken at the time of discectomy. There were 49 disc materials (from 38 men and 10 women [aged 19 to 78 years; average, 36.6 years]). The herniation was classified into four types, based on the intraoperative observations: protrusion (P), subligamentous extrusion (SE), transligamentous extrusion (TE), and sequestration (S). There were 19 P type discs, 3 SE type, 10 TE type, and 17 S type. The surgical specimens were stained with hematoxylin-eosin, as well as immunohistological staining with the labelled streptavidin biotin method, using human T-cell, human B-cell, and human macrophage antibodies. Inflammatory-cell infiltration was observed at the border of the disc. These findings were present in 19 discs (70%) of the 27 discs of TE and S types (10 TE and 17 S types), but were not seen in the 22 discs of P and SE types (19 P and 3 SE types). Immunohistological staining of the area with inflammatory-cell infiltration revealed the presence of T cells and macrophages, which suggested that this cell infiltration originated from T cells and macrophages, and that the spontaneous resorption of the disc may have resulted from the phagocytic activities of these cells.

Adult↗

Reconstruction of rabbit Achilles tendon with three bioabsorbable materials: histological and biomechanical studies.

This study investigated whether three biodegradable materials, poly-N-acetyl-D-glucosamine (chitin), poly-epsilon-caprolactone (p-CL), polylactic acid (PLA), and chitin/p-CL composite could be used as scaffold implants in the reconstruction of extra-articular ligaments or tendons. Braided artificial tendons made from these materials were soaked in phosphate-buffered saline or implanted in the subcutaneous tissue of 31 Japanese white rabbits and then subjected to load at failure testing. There was no significant loss of load at failure when chitin tendons were soaked in saline for 26 weeks, but a rapid decrease occurred in vivo. The other two types of tendons showed significant loss of strength after 26 weeks both in vitro and in vivo. Another 111 rabbits were used to assess Achilles tendon reconstruction with the braided tendon implants. The tendon regeneration process was assessed macroscopically, histologically, immunohistochemically, and mechanically (maximum load at failure). Chitin showed more rapid degradation than the other materials on histological examination. There was good formation of fibrous tissue composed of type I and type III collagen around the chitin, PLA, and chitin/p-CL fibers in comparison with p-CL fibers, but chitin tendons showed more rapid loss of strength after implantation. Both PLA and the chitin/p-CL composite tendons had good initial strength and showed increased ingrowth of fibrous tissue, suggesting that these materials are promising as artificial tendons.

Achilles Tendon↗

Progressive outer retinal necrosis caused by herpes simplex virus type 1 in a patient with acquired immunodeficiency syndrome.

OBJECTIVE/BACKGROUND: To identify the etiologic agent of rapidly progressive outer retinal necrosis (PORN) in a 32-year-old man with acquired immunodeficiency syndrome (AIDS), who had retinitis developed from cytomegalovirus (CMV). Multiple yellowish spots appeared in the deep retina without evidence of intraocular inflammation or retinal vasculitis, diagnosed clinically as PORN. Death occurred after failure of multiple organs. DESIGN: Case report. METHODS: Both globes were taken at autopsy, fixed in formalin, and examined histopathologically and immunohistochemically to identify causative agents in the retinal lesions. MAIN OUTCOME MEASURE: Immunohistochemistry. RESULTS: All layers of the retina were severely damaged and contained focal calcification. Cytomegalic inclusion bodies were found in cells in the damaged retina of the right eye. Immunohistochemical studies for herpesviruses revealed the presence of CMV antigens in the right retina at the posterior pole and herpes simplex virus type 1 (HSV-1)-specific antigen in the periphery of both retinas. No varicella-zoster virus (VZV) antigen was detected in either retina. CONCLUSIONS: PORN has been described as a variant of necrotizing herpetic retinopathy, occurring particularly in patients with AIDS. Although the etiologic agent has been reported to be VZV, HSV-1 can be an etiologic agent.

AIDS-Related Opportunistic Infections↗

Demonstration of diastolic and presystolic Purkinje potentials as critical potentials in a macroreentry circuit of verapamil-sensitive idiopathic left ventricular tachycardia.

OBJECTIVES: The purpose of this study was to determine the relation of diastolic and presystolic potentials recorded during verapamil-sensitive idiopathic left ventricular tachycardia (ILVT) to reentry circuit. BACKGROUND: Successful ablation of verapamil-sensitive ILVT at the zone of slow conduction from which the diastolic potential is recorded has been reported. However, the relationship between the diastolic potential and the reentrant circuit remains a matter of debate. METHODS: Radiofrequency (RF) ablation was performed in 20 patients with verapamil-sensitive ILVT. After identifying the ventricular tachycardia (VT) exit site, we searched for the mid-diastolic potential (P1) during VT. Entrainment followed by RF current application was performed. If the mid-diastolic potential could not be detected, RF current was applied at the VT exit site showing the earliest ventricular activation with a single fused presystolic Purkinje potential (P2). RESULTS: In 15 of 20 patients, both P1 and P2 were recorded during VT from midseptal region. Entrainment pacing captured P1 orthodromically and reset the VT. The interval from stimulus to P1 was prolonged as the pacing rate was increased. Radiofrequency ablation was successfully performed at this site in all 15 patients. After successful ablation, P1 appeared after the QRS complex during sinus rhythm with the identical sequence to that during VT. In the remaining five patients, the diastolic potential could not be detected, and a single fused P2 was recorded only at the VT exit site. Successful ablation was performed at this site in all five patients. CONCLUSIONS: This study demonstrates that P1 and P2 are critical potentials in a circuit of verapamil-sensitive ILVT and suggests the presence of a macroreentry circuit involving the normal Purkinje system and the abnormal Purkinje tissue with decremental property and verapamil-sensitivity.

Adolescent↗