Biomedical subjects

K Miyashita

Publications and source records attributed to K Miyashita.

At least 19 recordsLinked to original sources

PubMed1999

The chlorocatechol-catabolic transposon Tn5707 of Alcaligenes eutrophus NH9, carrying a gene cluster highly homologous to that in the 1,2,4-trichlorobenzene-degrading bacterium Pseudomonas sp. strain P51, confers the ability to grow on 3-chlorobenzoate.

Alcaligenes eutrophus (Ralstonia eutropha) NH9, isolated in Japan, utilizes 3-chlorobenzoate as its sole source of carbon and energy. Sequencing of the relevant region of plasmid pENH91 from strain NH9 revealed that the genes for the catabolic enzymes were homologous to the genes of the modified ortho-cleavage pathway. The genes from strain NH9 (cbnR-ABCD) showed the highest homology (89 to 100% identity at the nucleotide level) to the tcbR-CDEF genes on plasmid pP51 of the 1,2,4-trichlorobenzene-degrading bacterium Pseudomonas sp. strain P51, which was isolated in The Netherlands. The structure of the operon, including the lengths of open reading frames and intervening sequences, was completely conserved between the cbn and tcb genes. Most nucleotide substitutions were localized within and proximal to the cbnB (tcbD) gene. The difference in the chloroaromatics that the two strains could use as growth substrates seemed to be due to differences in enzymes that convert substrates to chlorocatechols. The restriction map of plasmid pENH91 was clearly different from that of pP51 except in the regions that contained the cbnR-ABCD and tcbR-CDEF genes, respectively, suggesting that the chlorocatechol gene clusters might have been transferred as units. Two homologous sequences, present as direct repeats in both flanking regions of the cbnR-ABCD genes on pENH91, were found to be identical insertion sequences (ISs), designated IS1600, which formed a composite transposon designated Tn5707. Although the tcbR-CDEF genes were not associated with similar ISs, a DNA fragment homologous to IS1600 was cloned from the chromosome of strain P51. The sequence of the fragment suggested that it might be a remnant of an IS. The two sequences, together with IS1326 and nmoT, formed a distinct cluster on a phylogenetic tree of the IS21 family. The diversity of the sources of these IS or IS-like elements suggests the prevalence of ISs of this type.

Amino Acid Sequence↗

PubMed1998

Visualization of pressure-dependent luxury perfusion in a patient with subacute cerebral infarction.

Luxury perfusion characterized by depressed metabolism compared with CBF might be changed by decreasing cerebral perfusion pressure during the sitting position. A 77-yr-old man with subacute cerebral infarction was studied with brain X-ray computed tomography (CT), raise-up test with 99mTc-d,1-hexamethylpropyleneamine oxime (HMPAO) brain single photon emission tomography (SPECT) and positron emission tomography (PET). Brain X-ray CT revealed a low-density area in the left middle cerebral artery (MCA) anterior area. Raise-up 99mTc-HMPAO brain SPECT revealed decreased uptake in the left MCA anterior area in the sitting position and subsequent supine 99mTc-HMPAO brain SPECT revealed hot accumulation there. PET study in the supine position demonstrated some differences between CBF and the cerebral metabolic rate for oxygen in the left MCA anterior area, indicating luxury perfusion. CBF in the area of luxury perfusion might be decreased during the sitting or standing position and increased during the supine position by dysautoregulation of the cerebral vessels in the luxury perfusion during the subacute infarct.

Aged↗

PubMed1998

Characteristics of a long-term procedural skill in the monkey.

The purpose of this study was to characterize the nature and structure of procedural memory. We have previously studied the process of learning sequential behavioral procedures using monkeys. The monkey's task was to press five consecutive pairs of buttons (indicated by illumination) in the correct order for every pair, which he had to find by trial-and-error in a block of trials. The whole sequence was called a "hyperset"; each pair was called a "set". We first examined whether monkeys learned to perform a hyperset as a single sequence or learned the order of button-presses individually for each set. To answer this question, we generated hypersets that were the same as the hypersets that had been extensively learned except that the order of the sets was reversed. The performance of these "reversed hypersets" was much worse than the performance of the original learned hypersets and was similar to the performance of new hypersets, as regards both the number of errors and the performance time. The result suggests that monkeys learned a hyperset as a sequence. To examine whether the learned performance was specific to the hand used for practice, we had monkeys use the same hand throughout the long-term practice of each hyperset, and then tested the opposite hand. The performance using the opposite hand was worse than the performance using the trained hand, but was better than the performance for new hypersets. This indicates that the memory for the sequential procedure is only partially accessible to the hand that was not used for the practice.

Animals↗

PubMed1998

Hearing impairment among young Chinese in an urban area.

To evaluate hearing levels among young Chinese in an urban area, audiometry was carried out in Jinan city, Shandong Province, China. The subjects were 442 healthy school children and students ranging in age from 6-19 y. All subjects were asked to complete a brief questionnaire on otological symptoms, personal histories and use of noisy playthings. Audiometric threshold testing was performed at the audiometric frequencies of 0.5, 1, 2, 4 and 8 kHz. Logistic regression analysis was used to analyse the items of personal histories related to hearing impairment. Forty-five subjects (10% of the subjects) were classified as abnormal. Nineteen ears of the abnormal subjects showed 4 kHz-dip and 38 ears showed high-frequency hearing impairment. Lower prevalence of hearing impairment was found when compared with young Chinese in a rural area (20%). The logistic regression analysis showed that head trauma may be the cause of hearing impairment among young Chinese in urban areas. Otological examinations for all children sustaining head trauma are recommended.

Adolescent↗

PubMed1998

Development of the selective adsorbent for EDA containing fibronectin using heparin immobilized cellulose.

We have noticed that extra domain A containing fibronectin (EDA(+)FN) is closely related to inflammatory diseases and accordingly investigated a selective adsorbent to remove EDA(+)FN, which may increase the severity of the symptoms, from the plasma of rheumatoid arthritis (RA) patients. Three types of heparin immobilized cellulose (HC) were prepared and their adsorptive properties were evaluated by batchwise adsorption tests. The results showed that EDA(+)FN was adsorbed more efficiently than total FN on HC. In particular HC in which heparin was immobilized to amino residue had an excellent capacity. The adsorption rates of EDA(+)FN, total FN and antithrombin-III were 89, 15 and 17% respectively. This adsorbent is extremely useful in selectively removing EDA(+)FN from plasma.

Adsorption↗

PubMed1998

Quantitation of herpes simplex viral DNA in Vero cells for evaluation of an antiviral agent using the polymerase chain reaction.

A method for quantitation of the DNA of Herpes simplex virus type 2 (HSV-2)-infected Vero cells by the polymerase chain reaction (PCR) was developed. This method allowed recognition of several molecules of viral DNA among the total DNA extracted from cells. The method could be applied to a very large range (10(-0)-10(-7)) of initial amounts of template. Products of PCR were collected after each cycle for kinetic analysis. Products were subjected to electrophoresis and amplified bands were stained with ethidium bromide. The intensity of fluorescence of each band was measured with a charge-coupled device (CCD) image analyzer. The time course of increases in the relative yield of viral DNA was determined. Two-fold amplification of viral DNA occurred each 6-h cycle from 7 h after infection. Using this method, the yields of viral DNA after treatment with the drug acyclovir (ACV) at 0.1 and 2 microg/ml were about 1/10 and 1/80 of those from nontreated infected cells, respectively. These results indicate that this method makes clear the inhibitory effect of ACV on the synthesis of viral DNA.

Acyclovir↗

PubMed1998

Denture mobility with six degrees of freedom during function.

The purpose of this study was (1) to assemble and verify a system to measure the three-dimensional (3-D) movement of the upper and lower complete dentures and the movement of the mandible simultaneously, and (2) to analyse the relation between denture movements and the path of closure of the mandible during function. A 3-D motion capture system with four infrared TV cameras was used for this purpose. The relation between the dentures and the mandibular movements was analysed through the change of the inner product of normal vectors of the denture occlusal planes and mandibular planes. The mandibular movements were classified into two types; the normal stroke (the path of closure was on the ipsilateral side of mastication) and the reverse stroke (on the contralateral side). The results showed that the system could measure the denture mobility within a 0.3 mm error. The mobility of the upper dentures had a correlation to the path of closure of the mandible regardless of the working side or nonworking side, and the lower dentures had a tendency to move toward the working side.

Aged↗

PubMed1998

Relevance of periglomerular myofibroblasts in progression of human glomerulonephritis.

To clarify the pathological and clinical significance of periglomerular alpha-smooth muscle actin (alpha-SMA)-positive cells, we examined 51 needle-biopsy specimens from patients with human glomerulonephritis. Immunoelectron microscopy confirmed these cells were myofibroblasts showing characteristic features with abundant alpha-SMA-positive thin myofilaments. Nonsclerotic glomeruli with periglomerular myofibroblasts were larger in the Bowman's capsular planar area than nonsclerotic glomeruli without periglomerular myofibroblasts (24.7 +/- 6.0 x 10(3) microm2 v 19.9 +/- 8.5 x 10(3) microm2; P < 0.01). We studied the correlation between the clinical prognosis and the extent of periglomerular myofibroblasts in 24 patients with IgA nephropathy. Patients were divided into two groups; those with plasma creatinine levels within normal range at biopsy and significantly elevated at follow-up were designated group 1 (poor prognosis), and patients with plasma creatinine levels within normal range at biopsy and not significantly elevated at follow-up were designated group 2 (fair prognosis). In the kidneys of group 1 patients, periglomerular alpha-SMA was expressed more intensively than it was in the kidneys of group 2 patients (alpha-SMA expression score, 1.0 +/- 0.48 v 0.52 +/- 0.54; P < 0.05). These findings indicate that periglomerular myofibroblasts appeared surrounding the nonsclerotic hypertrophic glomeruli, which may lead finally to glomerulosclerosis. This report suggests that interaction between the glomerular cells and the periglomerular myofibroblasts may have a role in the progression of glomerular diseases.

Actin Cytoskeleton↗

PubMed1998

Presymptomatic brain lesions on MRI in a patient with intravascular malignant lymphomatosis.

A 58-year-old man with a intravascular malignant lymphomatosis initially developed myeloradiculopathy without cerebral symptoms. His MRI, however, demonstrated solid, wedge-shaped, and well-demarcated lesions in the deep white matter and a string-shaped lesion along with nerve fibers in the splenium of corpus callosum. A variety of cerebral symptoms manifested a month afterward. The possibility of this disease should be considered in cases of undiagnosed myeloradiculopathy with such silent brain lesions.

Brain↗

PubMed1998

glkA is involved in glucose repression of chitinase production in Streptomyces lividans.

Chitinase production in Streptomyces lividans is induced by chitin and repressed in the presence of glucose. A mutant of S. lividans TK24, strain G015, which was defective in glucose repression of chitinase production, was obtained by screening colonies for zones of clearing on colloidal chitin agar plates containing 1.0% (wt/vol) glucose. The transcriptional analysis of chiA in G015 with xylE, which encodes catechol 2,3-dioxygenase, as a reporter gene showed that the transcription from the chiA promoter of S. lividans TK24 occurred regardless of the presence of glucose. G015 was resistant to 2-deoxyglucose (2-DOG) and did not utilize glucose as a sole carbon source. When a DNA fragment containing glkA, a gene for glucose kinase, of Streptomyces coelicolor A3(2) was introduced into strain G015 on a low-copy-number plasmid, the sensitivity to 2-DOG, the ability to utilize glucose, and the glucose repression of chitinase production were restored. These results indicate that glkA is involved in glucose repression of chitinase production in S. lividans TK24.

Bacterial Proteins↗

PubMed1998

Mesangial expression of a nonmuscle myosin heavy chain, SMemb, is associated with glomerular sclerosis and renal prognosis in IgA nephropathy.

To characterize the phenotypic alteration in mesangial cells in human glomerulonephritis, we investigated the expression of nonmuscle-type myosin heavy chain, SMemb, and alpha-smooth muscle actin (alpha-SM actin) in IgA nephropathy. The expression of SMemb and alpha-SM actin was examined by immunohistochemistry in biopsy specimens from 45 patients with IgA nephropathy. We examined a total of 489 glomeruli representing all patients enrolled, and found that mesangial expression of SMemb and alpha-SM actin was associated with mesangial proliferation. Only mesangial expression of SMemb showed a significant relationship with mesangial matrix accumulation. Semiquantitative evaluation using composite expression scores showed that the expression of SMemb was elevated in the patients with poor renal prognosis. The expression of alpha-SM actin showed no significant relationship with renal prognosis. These results suggest that mesangial expression of SMemb is an important factor in the progression of IgA nephropathy, and that SMemb and alpha-SM actin are associated with the activation of mesangial cells by different mechanisms.

Adolescent↗

PubMed1998

Enhanced in vitro percutaneous penetration of salicylate by ion pair formation with alkylamines.

The apparent octanol/water partition coefficient (APC) of salicylate (SA) increased as the concentration of alkylamine (amyl, hexyl, heptyl, octyl and nonylamine) in aqueous phase increased, presumably through intermolecular ion pair formation between the negatively charged SA moiety and the alkylamine cation. The true partition coefficient (TPC) and the formation constant (Kf) of the ion pair were calculated from the partition data. The skin permeability of SA increased as the APC of SA increased, when 20-fold molar excess of alkylamine was added to the donor compartment. Permeability of ion pairs (PAB) from the aqueous phase to a shed snake skin was estimated from the permeability data assuming 1:1 ion pair. The methylene group contribution to the free energy of transfer of ion pairs from water to the shed snake skin was less than the reported value for nonionized drugs. This suggests that the ion pair is more polar by nature than nonionized molecules, even if ionic characteristics are masked to some extent by ion pair formation.

Amines↗

PubMed1998

Health surveillance of forestry workers exposed to hand-arm vibration in Wakayama from 1974 to 1996.

To clarify the trends in the number and the severity of vibration syndrome in Wakayama for these 23 years, the records of the medical examinations for vibration syndrome were analyzed with 4,652 (a total of 9,920) private forestry workers exposed to hand-arm vibration. The number of the subjects who took the medical examinations reached a maximum of 1,242 in 1978. After that, it decreased year by year, but remained at about 300 or less from 1988. There was a corresponding increase in age and the number of years of operating chain saws among the subjects examined. The compensated cases reached a maximum number of 393 cases in 1977, and then a drastic decrease was noted. Year by year there was an increase in the number of cases whose medical examinations revealed no abnormality. The regional occupational health care system including the surveillance card, which has been active since 1975, has probably contributed to the decrease or prevention of occurrence of vibration syndrome in Wakayama.

Adult↗

PubMed1998

Finger skin temperature and laser-Doppler finger blood flow in subjects exposed to hand-arm vibration.

OBJECTIVES: This study was aimed to assess the peripheral circulatory functions of forestry workers exposed to hand-arm vibration, using simultaneous measurements of finger skin temperature (FST) and finger blood flow (FBF). METHODS: The subjects were five men with vibration-induced white finger (VWF), four subjects with numbness in their hands and five exposed controls without any signs or symptoms related to vibration syndrome. Simultaneous measurements of FST and laser-Doppler FBF were performed before, during and following a 10-min recovery period after the hand was immersed in cold water (10 degrees C, 10 min). Partitional measurements of finger blood flows could be made between the capillaries and deeply located vessels using two laser-Doppler flow probes having narrower and wider optic fiber separations. RESULTS: The FST as well as the FBF of the capillary and deeply located vessels during and following the cold immersion test in those with VWF and in the subjects with numbness had a slight increase immediately following the cold immersion and no tendency to be augmented as the time span increased. After the cold immersion test, the control group had significantly greater FST and FBF compared to other groups. Particularly, from the 5th to the 10th min after the cold provocation, the values of FBF of the deeply located vessels in those with VWF were significantly lower than the controls. During the course of observation, the FST had significantly linear relationship with the recorded capillary and deep FBF values, and that was much closely correlated with the deep FBF (i.e., P < 0.01 in each selected condition). The slope of the regression lines in the capillary were steeper and shallow in the deep FBF. CONCLUSIONS: In addition to the measurement of capillary FBF, more detailed and accurate information on the peripheral circulatory functions of vibration-exposed subjects can be obtained by including the assessment of the deep FBF.

Aged↗

PubMed1998

Evaluation of finger skin temperature by cold provocation test for diagnosis of vibration induced white finger (VWF).

Clinical data of workers (40-69 yrs) operating chain saws for a ten-year period from 1986 to 1995 were analyzed to assess the evaluation standard of finger skin temperature for a cold provocation test (10 degrees C 10 min). Screening points of finger skin temperature for screening 191 workers with a vibration induced white finger (VWF) were obtained from receiver operating characteristic (ROC) curves. The screening points at 5 min and 10 min after a cold provocation were approximately equal to 50th percentiles of 217 workers with no symptoms (NS group). The screening points of recovery rates at 5 min and 10 min after a cold provocation almost agreed with 50th percentiles in NS group. A new evaluation standard was prepared in reference to these screening points and finger skin temperatures by fraction in NS group. The new one will be useful for the health care of workers operating vibrating tools under present working conditions.

Adult↗

PubMed1997

Differential roles of monkey striatum in learning of sequential hand movement.

To study the role of the basal ganglia in learning of sequential movements, we trained two monkeys to perform a sequential button-press task (2x5 task). This task enabled us to examine the process of learning new sequences as well as the execution of well-learned sequences repeatedly. We injected muscimol (a GABA agonist) into different parts of the striatum to inactivate the local neural activity reversibly. The learning of new sequences became deficient after injections in the anterior caudate and putamen, but not the middle-posterior putamen. The execution of well-learned sequences was disrupted after injections in the middle-posterior putamen and, less severely, after injections in the anterior caudate/putamen. These results suggest that the anterior and posterior portions of the striatum participate in different aspects of learning of sequential movements.

Animals↗

PubMed1997

Oxidative stability of liposomes prepared from soybean PC, chicken egg PC, and salmon egg PC.

The oxidative stability of phosphatidylcholines (PCs) from soybean, chicken egg, and salmon egg in liposomes was compared with that in aqueous micelles. When each PC was oxidized in aqueous micelles, salmon egg PC was the most oxidatively stable, followed by chicken egg PC and soybean PC, however, no significant difference in the oxidative stability was apparent between chicken egg PC and salmon egg PC in liposomes. The main molecular species of soybean PC was 1,2-dilinoleoyl-PC, while most of the PUFAs in chicken egg PC and salmon egg PC were not esterified at the sn-1 position but at the sn-2 position. Therefore, it is suggested that the oxidative stability of PC liposomes would be strongly influenced by the positional distribution of PUFAs in the PC molecule. Further studies on the oxidation of PC liposomes showed that chicken egg albumin and soybean protein protected PC bilayers against attack by free radicals generated in the aqueous phase.

Animals↗

PubMed1997

Immunohistochemical and biochemical characterization of sulphated proteoglycans in embryonic chick bone.

The type and distribution of sulphated proteoglycans (PGs) in the midshaft subperiosteal bone of 15-18-day embryonic chick femurs were studied immunocytochemically and biochemically, using four monoclonal antibodies (MAb 2B6, 3B3, 1B5, and 5D4). These MAb specifically recognize epitopes in chondroitin 4-sulphate (C4-S) and dermatan sulphate (DS); chondroitin 6-sulphate (C6-S) and unsulphated chondroitin (C0-S); C0-S; and keratan sulphate (KS) respectively. Immunohistochemistry showed that staining of C4-S, DS, and KS, but not of C6-S and C0-S, was limited to osteoid, the cell surface of osteocytes, and to the walls of osteocytic lacunae and bone canaliculi in 15-18-day embryonic specimens. However, no significant difference in the distribution and intensity of immunostaining was observed in these specimens. Bone proteins were extracted from fresh 18-day embryonic specimens with a three extraction procedure, 4 M guanidine HCl (GdnCl, G-1 extract), 0.4 M EDTA (E-extract), followed by GdnCl (G-2 extract), to characterize mineral binding and collagenous matrix associated PGs in E- and G2-extracts respectively. Western blot analysis of E- and G2-extracts demonstrated that chondroitinase ABC-digested PGs with a molecular weight (Mr) approximately of 45,000 containing GAGs predominantly corresponding to C4-S and/or DS, with no detectable C6-S or C0-S present in the mineral and matrix phase, whereas KSPGs having an Mr of approximately 72,000 are only present in the mineral phase. These results indicate that embryonic chick bone contains small PGs having C4-S, DS, and KS chains with preferential localization to osteoid, the cell surface of osteocytes, and to the walls of osteocytic lacunae and bone canaliculi.

Animals↗