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Biomedical subjects

S Yamaguchi

Publications and source records attributed to S Yamaguchi.

At least 379 records · Page 21Linked to original sources

Viruses produced from complementary DNA of virulent and avirulent strains of swine vesicular disease viruses retain the in vivo and in vitro characteristics of the parental strain.

A full-length cDNA copy of the genome of the pathogenic strain, J1'73, of swine vesicular disease virus (SVDV) was constructed and inserted into the plasmid pSVL to generate a recombinant plasmid pSVLSJ1. Infectious virus was produced following transfection of cultured mammalian cells with the plasmid. The recovered virus had the same in vitro properties as the parental strain with regard to antigenicity, plaque size on IBRS-2 cells and single-step growth. Pigs were experimentally infected with the parental virus, J1'73 strain, and viruses recovered from cells transfected with the plasmids pSVLSJ1 and pSVLS00 [Inoue T, Yamaguchi S, Saeki T, Sekiguchi K, J Gen Virol 71: 1,835-1,838 (1990)] corresponding to the pathogenic (J1'73) and non-pathogenic (H/3'76) Japanese strains of the SVDV, respectively. All pigs inoculated with the virus recovered from pSVLSJ1 produced clinical signs of similar severity to those inoculated with the parental J1'73 strain. In contrast, pigs inoculated with the virus recovered from pSVLS00 did not show any clinical signs. Viruses recovered from cells transfected with either pSVLSJ1 or pSVLS00 therefore retained the in vitro characteristics and the in vivo pathogenicity of their respective parental strains.

Animals↗

Selective impairment of HCO3(-)-dependent pHi regulation by lysophosphatidylcholine in guinea pig ventricular myocardium.

OBJECTIVE: The aim was to examine the effects of lysophosphatidylcholine (LPC), an amphiphilic lipid metabolite in ischemic myocardium, on intracellular pH (pH(i)) regulatory systems in guinea pig papillary muscles. METHODS: In CO2/HCO(3-)-buffered Tyrode solution, pH(i), intracellular Na+ activity (aNai) and membrane potential of isolated guinea pig papillary muscles were measured using ion-selective microelectrode and conventional microelectrode. Standard ammonium prepulsing with 20 mM NH4Cl was used to produce an intracellular acid load, and effects of LPC on the pH(i) recovery from acidosis were evaluated in the absence and presence of a transport inhibitor. RESULTS: LPC acidified the resting pH(i) by 0.03 +/- 0.01 pH units (n = 15, p < 0.01) concomitantly with a slight decrease in resting membrane potential and an increase in aNai in quiescent preparations. The pH(i) recovery rate from an intracellular acid load was decreased to 83 +/- 4% of the control value by 30 microM LPC (n = 8, P < 0.05) but not by 30 microM phosphatidylcholine (PC). In the presence of 10 microM 5-(N,N-hexamethylene) amiloride (HMA), a Na(+)-H+ exchange inhibitor, LPC still slowed pH(i) recovery from an intracellular acid load to 77 +/- 4% of the control (n = 5, P < 0.05). However, LPC failed to alter the pH(i) recovery rate in the presence of 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS, 0.5 mM), a Na(+)-HCO3- symport inhibitor. CONCLUSION: LPC impairs Na(+)-HCO3- symport but not Na(+)-H+ exchange, and LPC may potentiate its arrhythmogenic action by intensifying the intracellular acidosis in ischemic myocardium.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗

Glutamate receptors: brain function and signal transduction.

Glutamate receptors are important in neural plasticity, neural development and neurodegeneration. N-methyl-d-aspartate (NMDA) receptors and alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA)/kainate receptors act as glutamate-gated cation channels, whereas metabotropic receptors (mGluRs) modulate the production of second messengers via G proteins. Molecular studies from our and other laboratories indicated that NMDA receptors and mGluRs exist as multiple subunits (NMDAR1 and NMDAR2A-2D) and multiple subtypes (mGluR1-mGluR8). In light of the molecular diversity of glutamate receptors, we explored the function and intracellular signaling mechanisms of different members of glutamate receptors. In the visual system, retinal bipolar cells receive glutamate transmission from photoreceptors and contribute to segregating visual signals into ON and OFF pathways. The molecularly cloned mGluR6 is restrictedly expressed at the postsynaptic site of ON-bipolar cells in both rod and cone systems. Gene targeting of mGluR6 results in a loss of ON responses without changing OFF responses and severely impairs detecting visual contrasts. Since AMPA receptors mediate OFF responses in OFF-bipolar cells, two distinct types of glutamate receptors effectively operate for ON and OFF responses. mGluR1 and mGluR5 are both coupled to inositol triphosphate (IP3)/calcium signal transduction with an identical agonist selectivity. Single-cell intracellular calcium ([Ca2+]i) recordings indicated that glutamate evokes a non-oscillatory and oscillatory [Ca2+]i response in mGluR1-expressing and mGluR5-expressing cells, respectively. This difference results from a single amino acid substitution, aspartate of mGluR1 or threonine of mGluR5, at the G protein-interacting carboxy-terminal domains. Protein kinase C phosphorylation of the threonine of mGluR5 is responsible for inducing [Ca2+]i oscillations in mGluR5-expressing cells and cultured glial cells. Thus, the two closely related mGluR subtypes mediate diverging intracellular signaling in glutamate transmission.

Animals↗

Anatomic study of the pisotriquetral joint: ligament anatomy and cartilagenous change.

Eighty embalmed cadaver wrists (40 right and 40 left, 55 male and 25 female, aged 36 to 93 years [average age, 72.5 years]) were dissected to assess the anatomy of the pisotriquetral joint and the location of degenerative changes. The patterns of degenerative changes were classified into five types: type 1, central (8.8%); type 2, peripheral (38.8%); type 3, fan-shaped (13.8%); type 4, mixed (combination of > or =1 of types 1, 2, and/or 3; 10.0%); and type 5, total (12.5%). There was no cartilagenous change in 16.2% of the pisoform and 13.7% of the triquetrum. Degenerative changes (exposed subchondral bone and chondromalacia) on the articular surface of the pisiform were present in 83.3% of the wrists and on the triquetrum in 86.3% of the wrists. Degenerative changes were most commonly of the type 2 pattern (peripheral), with change located most commonly in the distal, distal-radial, and radial aspects of the pisiform and triquetrum. The ligamentous anatomy was categorized into 3 different types: type A, pisohamate and pisometacarpal ligaments inserted on the palmar distal aspect of the pisiform (52.5%); type B, pisohamate ligament inserted on the radial side of the pisiform and pisometacarpal ligament inserted on the palmar side of the pisiform (41.3%); and type C, type B anatomy with an additional ligamentous slip between the pisometacarpal ligament and the distal aspect of the hook of the hamate (6.2%).

Adult↗

The carpal stretch test at the scapholunate joint.

The purpose of this study was to evaluate the principle of the carpal stretch test. Eleven upper extremities from 6 fresh cadaver specimens and 40 normal wrists from 20 volunteers were used in this study. Changes in intercarpal distances (carpal interval ratio of the radioscaphoid, radiolunate, and lunocapitate joints) and step-off deformities at the scapholunate joint seen on the posteroanterior view of an x-ray film were analyzed with and without the carpal stretch test. In the cadaver wrists, the dorsal capsulotomy and radioscapholunate ligament (stage 1), the palmar half of the scapholunate interosseous ligament (stage 2), the dorsal half of the scapholunate interosseous ligament (stage 3), and the radioscaphocapitate ligament (stage 4) were sequentially sectioned. The carpal stretch test was completed in each stage. Each carpal interval ratio increased as each stage was added. Step-off deformities at the scapholunate joint were seen in 6 of 11 wrists in stage 2 and in 10 wrists in stages 3 and 4 with the carpal stretch test. In the normal wrists, neither gender nor wrist laxity (Garcia's criteria) not influenced the results of the carpal stretch test. The carpal stretch test may be helpful in identifying occult and/or dynamic scapholunate dissociation.

Adult↗

3-Hydroxyisobutyric aciduria in two brothers.

Two Japanese brothers with 3-hydroxyisobutyric aciduria (3HiB-uria) are studied. The elder brother died of a ketoacidotic episode at the age of 4 years; the younger brother also manifested repeated episodes of ketoacidosis after 1 year of age. He is diagnosed as having 3HiB-uria by gas chromatography/mass spectometry analysis, using the unique fragment ions of 3HiB. Magnetic resonance imaging reveals focal white matter abnormalities. Protein restriction is effective for preventing the ketoacidotic episodes, although carnitine therapy seems less effective.

Child, Preschool↗

6-Phenylaliphatic-substituted androst-4-ene-3,17-diones as aromatase inhibitors: structure-activity relationships.

Two series of 6alpha- and 6beta-phenylaliphatic-substituted androst-4-ene-3,17-diones (3 and 5) were synthesized as aromatase inhibitors to gain insights of structure-activity relationships of varying the n-alkyl moiety (C2 to C5) of the 6-phenylaliphatic substituents to the inhibitory activity. All of the inhibitors synthesized inhibited human placental aromatase in a competitive manner with apparent Ki values ranging from 16 to 115 nM. The 6alpha-phenethyl analog 3a and the 6beta-phenbutyl analog 5c (Ki=16 nM for the two inhibitors, respectively) were the most potent inhibitors in each series. The inhibitory activities of the 6beta-substituted steroids 5 except for the phenethyl compound 5a were more powerful than those of the corresponding 6alpha-isomers 3. Elongation of the alkyl moiety of the 6-substituent of the 6alpha-phenethyl steroid 3a up to five methylene units decreased affinity to aromatase in all cases, whereas the addition of two more methylene units to the 6-side chain of the 6beta-phenethyl analog 5a increased the affinity in relation to carbon number of the 6-substituent. These results along with molecular modelling with the PM3 method, would give a new information about the formation of thermodynamically stable enzyme-inhibitor complex in a hydrophobic binding pocket in the active site of aromatase.

Androstenedione↗

Phase II study of paclitaxel (BMS-181339) intravenously infused over 3 hours for advanced or metastatic breast cancer in Japan. BMS-181339 Breast Cancer Study Group.

A Phase II study of paclitaxel in patients with primary advanced or metastatic breast cancer was conducted by a cooperative study group consisting of 16 institutions in Japan. Paclitaxel at a dose of 210 mg/m2 was intravenously infused over 3 hours, along with premedication to prevent hypersensitivity reactions. The course was repeated at 21-day intervals. Of 62 eligible patients, 60 were evaluable for toxicity and 59 were evaluable for efficacy. Forty-five patients were previously treated with anthracyclines. Twenty-one of 59 patients (35.6%) had a major objective response including 2 CRs and 19 PRs (95% confidence interval, 23.6-49.1%). A response rate of 35.5% (CR1, PR10) was observed in 31 patients refractory to the anthracyclines containing prior metastatic chemotherapy. Median (range) time was 41 (6-100) days to onset of and median duration of response was 125 (36-305) days. Toxicities included leukopenia (grade 3, 4: 67%), anemia (grade 1-3: 80%), thrombocytopenia (grade 1: 8%), alopecia (grade 3: 43%), peripheral neuropathy (grade 1-3: 93%), arthralgia (59%), myalgia (46%), nausea and vomiting (40%), fever (33%), allergic reaction (grade 3: 2%) and hypotension (grade 3: 5%). All toxicities were tolerable and manageable. Paclitaxel intravenously infused over 3 hours demonstrated a significant antitumor activity for metastatic breast cancer. Furthermore, paclitaxel exhibited non-cross resistance to anthracycline. Paclitaxel administered as a convenient 3-hour infusion is effective for patients with metastatic breast cancer and has an acceptable toxicity profile.

Adult↗

Detection of regional lymph node metastases in colon cancer by using RT-PCR for matrix metalloproteinase 7, matrilysin.

Lymph node metastasis is the most important prognostic factor in colon cancer. However, more accurate screening for metastasis than that afforded by conventional pathology remains elusive. We have employed a reverse transcriptase-polymerase chain reaction (RT-PCR) assay for a matrix metalloproteinase (MMP), 'matrilysin', because this gene is epithelial-specific and consistently expressed in colorectal cancer cells. The sensitivity of this assay was examined with the matrilysin-producing rectal cancer cell line 'CaR-1'. Matrilysin mRNA was detected in this system when more than 10(4) matrilysin-positive cells existed in a lymph node of ordinary size. Fourteen of 15 (93%) primary colon cancers and none of the surrounding normal tissues expressed matrilysin. All 10 histologically-positive lymph nodes were positive for matrilysin, while of 60 histologically-negative lymph nodes, eight were positive for matrilysin. When the additional sequential sectioning and histological re-examination was performed on five of these eight 'matrilysin-positive, but histologically-negative' lymph nodes, micrometastases were detected in three. Only one of the lymph nodes that were histologically-positive, but negative by matrilysin assay was from a patient with colon cancer in which matrilysin was not detected. In conclusion, RT-PCR assay for matrilysin is a sensitive method for detecting occult metastases in patients with colon cancer, and may complement histologic examination.

Adult↗

Localization of Fas and Fas ligand in bone marrow cells demonstrating myelodysplasia.

Frequent apoptosis in the bone marrow of patients with myelodysplastic syndromes (MDS) was demonstrated on frozen sections using the terminal deoxytransferase (TdT)-mediated dUTP nick end labeling (TUNEL) method. The overall mean percentage of TUNEL-positive cells was about 17% in the bone marrow of MDS, while bone marrow from control cases exhibited a mean of 3.4% (P < 0.001). To elucidate the mechanism of apoptosis in bone marrow cells of MDS, the expression of Fas antigen and Fas ligand (FasL) was examined by RT-PCR and immunohistochemistry. All MDS cases showed expression of Fas mRNA (12/12) and most exhibited an expression of FasL mRNA (10/12) by RT-PCR. Basically, control cases did not show positive signals for Fas and FasL mRNA, however, a very weak band was detected in three cases (3/10) for Fas and in one case (1/10) for FasL mRNA by RT-PCR. Immunohistochemical examination revealed positive staining for Fas (11/12) and FasL (12/12) in the bone marrow of MDS, while all the bone marrow samples from control cases were negative for anti-Fas (0/15) and for anti-FasL (0/15) antibody. Double staining clarified that TUNEL-positive apoptotic cells expressed Fas antigen on the cell surface, although not all Fas-positive cells were TUNEL positive. The Fas-positive cells of MDS bone marrow included hematopoietic cells expressing CD34 antigen, neutrophil elastase, a marker for myeloid series of cells, or glycophorin A, a marker for erythroid cells. However, CD68-positive cells which were macrophage lineage cells, did not express Fas antigen strongly. In contrast, positive staining for FasL was detected in hematopoietic cells and CD68-positive cells in the bone marrow of MDS. These results suggest that the Fas-FasL system plays an important role in inducing apoptosis in the bone marrow of MDS and works in an autocrine (hematopoietic cell-hematopoietic cell interaction) and/or paracrine (hematopoietic cell-stromal cell interaction) manner.

Adult↗

Flt-1, a receptor for vascular endothelial growth factor, has transforming and morphogenic potentials.

A paradox of Flt-1, a tyrosine kinase receptor for vascular endothelial growth factor (VEGF), is that the ligand cannot activate the receptor to stimulate growth of cells that exogenously overexpress the receptor. In order to find Flt-1 kinase-dependent biological systems, we obtained for the first time activated forms of the Flt-1 kinase in a ligand-independent manner. Replacement of the ABL sequences in the human leukemia oncoprotein BCR-ABL with the cytoplasmic domain of Flt-1 (BCR-FLT) followed by a retroviral random mutagenesis scheme gave constitutively active artificial chimera BCR-FLTm with mutations within the Flt-1 sequence. Like BCR-ABL it could, but not the original BCR-FLT, transform Rat1 fibroblasts, abrogate cytokine dependence in Ba/F3 cells, and induce neurite-like structures in neuronal PC12 cells. Interestingly, Rat1 cells transformed by BCR-FLTm formed tube-like structures in basement membrane matrix. BCR-FLTm retroviruses may be a very useful tool to investigate an as yet uncovered functions of the Flt-1 kinase.

Animals↗

Flagellar filament elongation can be impaired by mutations in the hook protein FlgE of Salmonella typhimurium: a possible role of the hook as a passage for the anti-sigma factor FlgM.

Among motile revertants isolated from flagellar hook-deficient (flgE) mutants of Salmonella typhimurium, one produced only short flagellar filaments in L broth, despite the fact that flagellin itself has the ability to polymerize into long filaments in vitro. This pseudorevertant has an intragenic suppressor, resulting in a two-amino-acid substitution (Asp-Gln-->Ala-Arg) in the C-terminal region of the hook protein, FlgE. The flagellation of the pseudorevertant was greatly affected by the concentration of NaCl in the culture media: we observed no filaments in the absence of NaCl, short filaments in 1% NaCl and full-length filaments in 2% NaCl. Electron microscopy of osmotically shocked cells showed that the number of hook-basal bodies on cells was constant under various NaCl conditions. Furthermore, we found that the mutant hook was straight rather than curved. We monitored the cellular flagellin level of this pseudorevertant under various NaCl concentrations by immunoblotting. It was revealed that little flagellin was present under NaCl-free conditions in contrast with the ordinary amounts of flagellin present in 2% NaCl. As the expression of flagellin is regulated by competitive interaction of a sigma factor, FliA, and a corresponding anti-sigma factor, FlgM, we also observed the effect of NaCl on the secretion of FlgM. FlgM was secreted into the media in more than 1% NaCl but accumulated inside the cells in the absence of NaCl, indicating that the failure of secretion of FlgM in the absence of salt was the cause of the impaired elongation of filaments.

Bacterial Proteins↗

Expression of Tamm-Horsfall protein in stone-forming rat models.

OBJECTIVES: To examine the expression of Tamm-Horsfall protein (THP) and calcium oxalate deposition in three rat models to clarify whether THP plays an active role in crystal formation or whether crystals induce the secretion of this protein. MATERIALS AND METHODS: A stone-forming rat model (model 1) with marked tubular dilatation in an entire kidney was produced by rendering Wistar rats (aged 8 weeks) hyperoxaluric and hypercalciuric, through compulsorily feeding with 0.12 mL of 5% ethylene glycol (in two doses daily) and 0.5 microgram of vitamin D3 every other day. Two other rat models were also produced. Model 2 comprised stone-forming rats with minimal tubular dilatation, achieved by giving rats the same dose of ethylene glycol once daily, and model 3 comprised stone-free rats with marked tubular dilatation achieved by unilateral ureteric ligation. The rats' kidneys were resected after 4 weeks and all resected kidneys immunohistochemically stained with an antibody to THP. Simultaneously, the location of calcium oxalate (CaOx) crystals was established with von Kossa staining. The relation between crystals and the secretion of THP was also assessed in vitro. Cultured renal epithelial cells (NRK-52E) were stained with an antibody to THP after they had been cultured for 72 h in a medium containing CaOx crystals. RESULTS: In model-1 kidneys with both tubular dilatation and many crystals, there was local and intense expression of THP in many renal tubules. CaOx crystals and the intense expression of THP tended to occur in the same renal tubules. In model 2 kidneys with little tubular dilatation, only a few renal tubules expressed THP strongly and the location of the crystals rarely coincided with that of THP expression. In model 3 kidneys with marked tubular dilatation but no crystals, THP was expressed strongly in many renal tubules. The expression of THP in cultured NRK-52E cells was not stimulated by CaOx crystals. CONCLUSIONS: The results from the in vivo models suggest that THP did not initiate crystal formation and the strong expression of THP was induced not by crystals but by renal tubular damage caused by tubular dilatation. From the close association of THP and crystals in model 1 kidneys, this protein might play a secondary role as an adhesive, promoting stone formation.

Adjuvants, Immunologic↗

Regional expression and regulation of alternative forms of mRNAs derived from two distinct transcription initiation sites of the rat mGluR5 gene.

Metabotropic glutamate receptor (mGluR) subtype 5 is expressed in both neuronal and glial cells and is thought to play an important role in neuronal plasticity. This expression is up-regulated during the early postnatal period and is induced in cultured astrocytes by specific growth factors. To investigate the mechanism underlying the regulation of mGluR5 expression, we isolated and characterized genomic clones containing the 5'-upstream exons and their flanking regions of the mGluR5 gene. On the basis of the mGluR5 genomic structure, cDNA recloning of the 5'-extreme region of mGluR5 as well as primer extension analysis indicated that mGluR5 mRNA is generated from two alternative first exons, termed exon 1A and exon 1B, which are separated by 1,949 bp and then connected to the common exon 2. Northern blot and in situ hybridization analyses indicated that two distinct transcription initiation sites are commonly used in the expression of mGluR5 mRNA in various, but specialized, brain regions and that these two alternative forms of mGluR5 mRNA are similarly up-regulated or down-regulated during the early postnatal period, depending on the brain regions. The two mRNAs are also expressed in cultured astrocytes but respond differently to growth factor-mediated induction. This study provides the genetic basis indicating the diverse mechanisms involved in the regulation of mGluR5 expression.

Age Factors↗

A case of microphthalmos with cyst and partial trisomy 22.

A 13-day-old girl was referred for evaluation of a right orbital mass. Ophthalmic examinations revealed microphthalmia with cyst in the right eye and microphthalmia in the left eye. She had lowset ears, auricular fistula, micrognathia, and muscular hypotonia. Chromosomal analysis using fluorescence in-situ hybridization (FISH) showed partial trisomy 22. To our knowledge, microphthalmos with cyst associated with partial trisomy 22 has not been described previously in the English literature. Repeated aspiration resulted in a decrease in size of the cyst.

Chromosomes, Human, Pair 22↗

A normative, community-based study of Mini-Mental State in elderly adults: the effect of age and educational level.

We investigated community-based data of the Mini-Mental State Examination (MMSE) scores of elderly residents along with the effects of age and educational level. MMSE was planned for all residents over 65 years of age in a town in northern Japan. The number of elders who took the MMSE was 2,266 (90%). The score significantly declined with age and lower educational level, although no effect of sex was apparent. For the MMSE subitems, all the values except for that of naming showed effects of both age and educational level. Those screened by MMSE who fell in the range of cognitive impairment (< 24) accounted for 21.8% and those with severe cognitive impairment (< 18) constituted 6.0%. Despite the differences in language and culture, the mean scores are remarkably similar between Japan and other countries. This is the first normative, community-based study of MMSE among elderly adults in Japan.

Aged↗

In vitro and in vivo selectin-blocking activities of sulfated lipids and sulfated sialyl compounds.

There is accumulating evidence that sulfated lipids, sulfated oligosaccharides and other sulfated compounds are reactive with selectins in a manner that interferes with selectin interactions with their natural ligands. In the report we describe the ability of sulfated lipids (sulfatides and gangliosides) and multimeric forms of sulfated sialic acid to block binding of P- and E-selectin-Ig to neutrophils. The in vivo ability of these compounds to block lung injury in rats following i.v. infusion of purified cobra venom factor (CVF), which induces injury that is L- and P-selectin dependent, was also determined as well as effects on recruitment of neutrophils, as measured by lung myeloperoxidase. There was a significant correlation between the ability of sulfated lipids and sialyl compounds to interfere in vitro with P-selectin-Ig binding to neutrophils and to protect against P-selectin-dependent acute lung injury induced by CVF. The biological effects of these sulfated compounds were also associated with diminished accumulation of neutrophils. The protective effects of these compounds may be linked to their ability to interfere with P-selectin binding to counter-receptors on neutrophils.

Animals↗