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Biomedical subjects

Y Yoshida

Publications and source records attributed to Y Yoshida.

At least 937 records · Page 52Linked to original sources

The 4 beta-methyl group of substrate does not affect the activity of lanosterol 14 alpha-demethylase (P-450(14)DM) of yeast: difference between the substrate recognition by yeast and plant sterol 14 alpha-demethylases.

Interaction of obtusifoliol and 24,28-dihydroobtusifoliol with yeast lanosterol 14 alpha-demethylase (P-450(14)DM) was studied to elucidate the role of the 4 beta-methyl group of substrate. P-450(14)DM of Saccharomyces cerevisiae catalyzed 14 alpha-demethylation of obtusifoliol. Apparent Vmax of obtusifoliol demethylation (15.4 nmol/min/nmol P-450) was similar to that of 24-methylene-24,25-dihydrolanosterol demethylation and was a little higher than those of lanosterol and 24,25-dihydrolanosterol demethylations. Apparent Km for obtusifoliol (12.0 microM) was higher than those for lanosterol and 24-methylene-24,25-dihydrolanosterol but was lower than that for 24,25-dihydrolanosterol. 24,28-Dihydroobtusifoliol was a very poor substrate for yeast P-450(14)DM. These facts suggest that the 4 beta-methyl group of sterol slightly affects the activity of yeast P-450(14)DM, while hydrogenation of a double bond in the sterol side-chain considerably impairs the activity. This finding is a contrast to the fact that the plant P-450(14)DM could not catalyze demethylation of sterols having 4 beta-methyl group, but favorably interacts with sterols having saturated side chain.

Cholestadienols↗

Effects of various bile acids and their sodium salts on development of pepsinogen-altered pyloric glands in rats.

Effects of dietary bile acids and their sodium salts on the development of pepsinogen-altered pyloric glands (PAPG) were examined in male WKY/N Crj rats initially given a single dose of 160 mg/kg body weight of N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) by gastric intubation. From week 3 the animals were administered basal diet containing 0.5% supplements of cholic acid (CA), deoxycholic acid (DCA), chenodeoxycholic acid (CDCA) or their sodium salts (Na-C, Na-DC and Na-CDC), or 5% ascorbic acid (ASA) or its salt (Na-AS) for 18 weeks. The concentration of DCA and Na-DC was reduced to 0.3% from week 12. At week 20, animals were killed and the numbers of immunohistochemically-demonstrated PAPG were determined. Values were significantly higher with Na-C and Na-CDC than with the corresponding parent acids, and in the Na-C case PAPG development was greater than with MNNG alone. In addition, Na-CDC itself induced the numbers of PAPG significantly. These results suggest that bile salts are possible intrinsic promoters of gastric carcinogenesis. They were without effect, however, on forestomach lesions.

Animals↗

N-nitrosoheptamethyleneimine-induced pulmonary and esophageal carcinogenesis and effects of concomitant treatment with bleomycin in rats.

The combination effects of bleomycin with N-nitrosoheptamethyleneimine (NHMI) or dihydroxy-di-N-propylnitrosamine (DHPN) on pulmonary carcinogenesis were investigated. Male F344 rats were given NHMI (20 or 40 ppm) or DHPN (200 ppm) in the drinking water and intraperitoneally injected with bleomycin (1 mg/kg) once a week for 18 weeks and then killed at week 24. Many rats treated with NHMI died before the termination of the experiment due to toxicity or development of advanced esophageal carcinomas, considered to be the main cause of death. Detailed histological examination performed on rats killed at week 24 revealed no statistically significant effects of bleomycin on NHMI or DHPN induction of neoplastic lesions in the lung or esophagus, although pulmonary carcinomas were only found in two rats treated with NHMI plus bleomycin. Under the present experimental conditions, NHMI exerted stronger carcinogenic activity in the esophagus than in the lung, and no obvious modifying effects of simultaneously administered bleomycin were evident on NHMI- or DHPN-induced pulmonary carcinogenesis.

Adenoma↗

Effect of acyl derivatives of 4,4,4-trifluoro-1-phenyl-1,3-butanedione on 2-acetylaminofluorene-induced glutathione S-transferase positive foci in the rat liver.

Acyl derivatives of 4,4,4-trifluoro-1-phenyl-1,3-butanedione (TFPB), 1-benzoyl-2-trifluoromethyl-2-acetoxyethene (BTAE), and 1-benzoyl-2-trifluoromethyl-2-(4-methylthio)benzoyloxyethene (BTME), were synthesized and investigated for inhibition of tRNA binding by N-acetoxy-2-acetylaminofluorene (N-AcO-AAF), and induction of glutathione S-transferase placental form (GST-P) positive foci in the rat liver by 2-acetylaminofluorene (2-AAF). Male F344 rats were given BTAE or BTME intraperitoneally and 2-AAF by intragastric intubation. Two weeks following the treatment, the rats were maintained on the diet containing 0.05% phenobarbital for an additional 6 weeks and then killed. Development of GST-P positive foci was not affected by concomitant treatment with BTAE or BTME. These two compounds inhibited the in vitro binding of N-AcO-AAF to tRNA. Thus, although these diacylmethane derivatives had the in vitro inhibitory activity, they did not inhibit tumor-initiating activity of 2-AAF in the rat liver.

2-Acetylaminofluorene↗

DNA ploidy of primary hepatocellular carcinoma and pulmonary metastases.

To better comprehend the differences in deoxyribonucleic acid (DNA) content between a primary hepatocellular carcinoma (HCC) and pulmonary metastatic nodules, tissue specimens taken at autopsy of 25 patients who had not received any drugs to treat the malignancy were examined using microspectrophotometry. The DNA distribution patterns were classified into Types I-III, and low (Types I and II) or high (Type III) ploidies, according to DNA distribution. Changes in the DNA content from high to low ploidies, namely DNA ploidy reduction from the primary lesion to pulmonary metastatic lesions, was evident in 9 of the 25 patients (36%), and changes from low to high were noted in 2 of the 25 patients (8%). The remaining 14 (56%) showed no evidence of changes in the DNA ploidy pattern. Reduction of DNA ploidy seen in HCC and its metastatic lesions in the lung may be one of the aspects of clonal evolution or selection mechanisms during the progression of tumor growth and metastasis.

Autopsy↗

An experimental study on the treatment of hydrofluoric acid burns.

A 20% solution of hydrofluoric acid (HF) was applied to the skin of rats and a biomedical observation of the tissues and sera was made. Flushing with running water was effective for HF burns. By applying 2.5% calcium gluconate jelly, concentrations of fluoride in the urine and the tissues surrounding the injured region were reduced. Thus, the results proved that irrigation with running water and jelly applications were evaluated as the most effective therapy among various methods tested for HF burns.

Animals↗

Age variation in the upper limit of hearing.

The upper limit of hearing was measured in 6105 otologically normal ears of subjects ranging in age from 5 to 89 years. The results are as follows: in each age group from 5 to 59 years in both sexes, the upper limit of hearing showed an approximately normal distribution if a logarithmic scale was used for the upper limit of hearing axis. The mode of the distribution shifted to a lower frequency with increasing age. Over age 60 years, the distribution became much wider. Standard upper limit age curves were established by calculating 10th, 25th, 50th, 75th and 90th percentiles for each age group. From early childhood where no age variation was recognized in conventional audiometry, deterioration of the upper limit of hearing was already in progress. This deterioration was slight between ages 25 and 39 but at ages over 40 it was accelerated and led to so-called presbycousis. The upper limit of hearing was found to be one of the best parameters for showing the quantitative age-related changes in hearing.

Adolescent↗

Human glial fibrillary acidic protein (GFAP): molecular cloning of the complete cDNA sequence and chromosomal localization (chromosome 17) of the GFAP gene.

We isolated three glial fibrillary acidic protein (GFAP) cDNA clones from a glioma cell line, U-251 MG. One clone isolated from a U-251 MG cDNA library was long, but lacked both ends. Using poly(A)+ RNA and primers synthesized according to the sequence of this clone, we used the polymerase chain reaction-assisted rapid amplification of cDNA ends (PCR-RACE) method, which is a strategy to isolate cDNA ends, and obtained cDNA clones for the 5' and 3' ends. From the sequences of these overlapping clones, the complete nucleotide sequence of human GFAP cDNA was established. The start (ATG) and the stop (TGA) signals were seen at nucleotide positions 15 and 1311, respectively, and divided the entire sequence of 3027 bp into 14 bp of 5' non-coding, 1296 bp of coding and 1717 bp of 3' non-coding regions. Using cDNA probes made from both the coding and the 3' non-coding regions, Northern blot hybridization was performed with two different stringencies on RNAs from human and rodent brains and human GFAP-positive and -negative cells. It was shown that the 3' non-coding region probe was more specific for human GFAP than the coding region probe which was specific only under higher stringency conditions. This was also suggested by homology analysis of the sequence with those of various intermediate filament proteins. Based on these findings, we performed spot blot hybridization of sorted human chromosomes and Southern blot hybridization of PCR-amplified DNAs of a panel of hamster-human somatic cell hybrids and localized the human GFAP gene to chromosome 17.

Amino Acid Sequence↗

Serum fluoride as an indicator of occupational hydrofluoric acid exposure.

To define the relationship between ionic fluoride concentration in the serum of workers and the amount of hydrofluoric acid (HF) in the work environment, pre-and postshift serum and urine samples of 142 HF workers and 270 unexposed workers were examined. The maximum and minimum concentrations of HF in the air in each workshop varied from the mean by less than 30%. The preexposure levels of serum and urinary fluoride in HF workers were higher (P < 0.001) than the control values. This suggests that fluoride excretion from the body continues for at least 12h. The postshift serum and urinary fluoride concentrations of these workers were significantly higher (P < 0.001) than the preshift concentrations. A good correlation (r = 0.64) was obtained between postshift serum fluoride and postshift urine fluoride. There was a linear relationship between mean serum fluoride concentration and HF concentration in the workshop. A mean fluoride concentration of 82.3 micrograms/l with a lower fiducial limit (95%, P = 0.05) of 57.9 micrograms/l was estimated to correspond to an atmospheric HF concentration of 3 ppm. This is the maximum allowable environmental concentration recommended by the Japanese Association of Industrial Health, and it is also the threshold limit value suggested by the American Conference of Governmental Industrial Hygienists. The results demonstrate that exposure to HF can be monitored by determining the serum fluoride concentration.

Adolescent↗

Distribution of 72-kDa heat-shock protein in rat brain after hyperthermia.

The distribution of the 72-kDa heat-shock protein (hsp72) in rat brain, 24 h following in vivo transient hyperthermia (41.5 degrees C, 15 min), was studied using immunohistochemistry (n = 22). Tissue sections were also stained with hematoxylin and eosin, and with an anti-glial fibrillary acidic protein to evaluate neuronal and astrocytic response to transient hyperthermia, respectively. hsp72 was observed in glia and endothelial cells throughout brain. hsp72 was also found in neurons located in the: dentate gyrus, habenula, and hypothalamus, granular layer of the cerebellum and the olfactory area. Our data indicate, that hyperthermia causes neuronal expression of hsp72, particularly in cerebral neuronal populations which control the neuroendocrine stress response.

Animals↗

Ultrastructural localization of calcium in mechanoreceptors of the oral mucosa.

Cytochemical localization of Ca2+ in Meissner corpuscles and Merkel cell-neurite complexes in the palatine mucosa of the Mongolian gerbil was studied by a combined oxalate antimonate-microwave irradiation procedure. The reaction products obtained were identified as calcium antimonate by EGTA solubility and X-ray microanalysis. Meissner corpuscles in the normal palatine rugae could be roughly classified into three types by amount and localization of Ca2+. Type I corpuscles were characterized by a high Ca2+ content in both the terminal axoplasm and caveolae of the lamellar plates, type II, by a low Ca2+ content in the terminal axoplasm and a high Ca2+ content in the lamellar cytoplasm. Type III corpuscles showed intermediate characteristics. Palatine rugae stimulated mechanically during fixation contained an increased number of type I corpuscles. On the other hand, two patterns were distinguished in the distribution of Ca2+ in Merkel cells in palatine rugae fixed under normal conditions. One showed abundant Ca2+ dispersed throughout the cell, while in the other, Ca2+ was specifically localized in the Golgi apparatus and mitochondria. Similar distribution patterns also were observed in palatine rugae that had received mechanical stimulus during fixation. Axon terminals of most Merkel cell-neurite complexes in normal palatine rugae were poor in axoplasmic Ca2+, whereas those in most Merkel cell-neurite complexes in mechanically stimulated palatine rugae contained abundant Ca2+ in their axoplasm.

Animals↗

Clinical features of long-term survivors of refractory myelodysplastic anemias. A Japanese cooperative study.

Eighty-one of 473 patients with refractory myelodysplastic anemias registered by the Japanese Cooperative Study Group prior to January 1987 survived more than 5 years. At presentation, most patients had mild cytopenia, a less aggressive form of the disease (48 with refractory anemia, 23 with refractory anemia with ring sideroblasts, 10 with refractory anemia with excess of blasts), less blast cells in the marrow and blood, and onset at younger ages. Their clinical profiles 5 years after presentation showed no significant improvement. The results suggest that the long-term survivors were found in a subpopulation of patients with a favorable prognosis and that it is a part of the natural course rather than the results of treatment.

Adolescent↗

On-line determination and resolution of the enantiomers of ketoprofen in plasma using coupled achiral-chiral high-performance liquid chromatography.

High-performance liquid chromatography (HPLC) using a column-switching technique has been applied to the on-line determination and resolution of the enantiomers of ketoprofen (KPF) as an acidic model compound. The system incorporates a mobile phase conversion stage (LC-2), which has a dilution tube and a trapping column, between the achiral chromatography stage (LC-1) and the chiral chromatography stage (LC-3). KPF in plasma was separated from plasma components and was determined with the aid of an internal standard in LC-1. The eluate containing KPF was selectively transferred to LC-2, where the eluate was adequately diluted with a new mobile phase by using the dilution tube to reduce the influence of the mobile phase from LC-1, and KPF was reconcentrated on the trapping column. Then the KPF enantiomers were resolved in LC-3 after column-switching. This system is accurate and rapid compared with conventional HPLC. Very high sensitivity could be achieved with our system when a microbore ovomucoid column was employed in LC-3. Incorporation of LC-2 allows the most favourable mobile phases for LC-1 and LC-3 to be used independently. This method is greatly superior to usual column-switching HPLC.

Chromatography, High Pressure Liquid↗

Mechanisms of relaxation induced by prostaglandins in isolated canine uterine arteries.

OBJECTIVE: Prostaglandins liberated from the uterus in response to chemical and physical stimuli would be important modulators of uterine arterial tone and blood flow. This study was aimed at analyzing mechanisms of vasodilator action of prostaglandins in uterine arteries. STUDY DESIGN: Canine uterine artery strips were suspended in Ringer-Locke solutions for isometric tension recording. RESULTS: Prostaglandin F2 alpha-induced relaxation was reversed to contraction by cyclooxygenase inhibitors and suppressed by tranylcypromine (prostaglandin I2 synthesis inhibitor) or diphloretin phosphate (an inhibitor of prostaglandin F2 alpha and prostaglandin I2 actions) but was unaffected by endothelium denudation. Prostaglandin E2-induced relaxation was not attenuated by indomethacin but was partially inhibited by a nitric oxide synthase inhibitor and endothelium denudation. Relaxation induced by beraprost (prostaglandin I2 analog) was suppressed by diphloretin phosphate but was not influenced by indomethacin and endothelium denudation. CONCLUSION: It appears that prostaglandin F2 alpha-induced relaxation is mediated by prostaglandin I2 released from subendothelial tissues, whereas prostaglandin E2-induced relaxation is caused by release of endothelium-derived nitric oxide and also by an endothelium-independent mechanism.

Animals↗

Dysregulation of adult T-cell leukemia-derived factor (ADF)/thioredoxin in HIV infection: loss of ADF high-producer cells in lymphoid tissues of AIDS patients.

Adult T-cell leukemia (ATL)-derived factor (ADF) is a multifunctional protein homologous to thioredoxin (TRX) with co-cytokine and thiol-dependent reducing activities. ADF/thioredoxin production is enhanced in T cells transformed by HTLV-I. We have examined the effect of HIV-1 infection on ADF/TRX expression using specific antibody against ADF/TRX. Lymph nodes from 5 AIDS and 1 AIDS-related complex (ARC) patients were examined. As a control, 8 HIV noninfected lymph nodes, including 3 cases with hyperplasia, were also examined. Immunohistopathological studies using normal HIV noninfected lymph nodes showed that ADF/TRX high-producer (ADFh) cells were macrophages and cells with dendritic morphology in the paracortical area. Abundant ADFh cells were observed in HIV noninfected hyperplastic lymph nodes. The number of ADFh cells was low in hyperplastic lymph nodes from an ARC patient. All of the lymph nodes of 5 AIDS cases were atrophic and the number of ADFh cells were extremely low. To verify these histochemical studies, we examined the effect of in vitro HIV infection on ADF/TRX expression in HTLV-I (+) T-cell lines. Western blot analysis showed that a reduction of ADF/TRX in HIV-1-infected SKT-1B and MT-2 cells, and the reduction inversely correlated with p24 antigen level. On the basis of the above in vivo and in vitro findings, we imply that the levels of ADF/TRX were down-regulated by HIV-1 infection and that the down-regulation may play a role for pathophysiology of HIV-infected individuals.

AIDS-Related Complex↗

Plasma membrane Ca(2+)-pump ATPase is not a substrate for cGMP-dependent protein kinase.

A plasma membrane Ca(2+)-pump ATPase preparation purified from porcine aorta was incubated with cGMP-dependent protein kinase (G-kinase) under the conditions under which dose-dependent stimulation of the enzyme by G-kinase was observed. Several proteins were phosphorylated, but two isoforms of plasma membrane Ca(2+)-pump ATPase with molecular masses of 135- and 145-kDa were not phosphorylated. The protein that was phosphorylated by G-kinase and identified in our previous study as the 135-kDa isoform of Ca(2+)-pump ATPase, on the basis of its almost identical mobility on SDS-PAGE, was found to be another protein with a molecular mass of 138 kDa. Fractionation of the enzyme preparation after incubation with G-kinase by a newly developed calmodulin affinity chromatographic method resulted in the separation of all the G-kinase substrates from the two isoforms of plasma membrane Ca(2+)-pump ATPase. These results suggest that the direct phosphorylation of the Ca(2+)-pump ATPase does not occur in association with the stimulation of the plasma membrane Ca(2+)-pump ATPase by G-kinase.

Animals↗

Photoactivated Photofrin II: astrocytic swelling precedes endothelial injury in rat brain.

Light activation of circulating hematoporphyrin derivatives has been used in the treatment of selected brain tumors. The effects of this photodynamic therapy on the non-neoplastic, adjacent brain tissue are incompletely characterized. We studied in adult Fisher rats the time-dependent (1 hour to 7 days) effects of photoactivated Photofrin II. Our protocol was comparable to that used in the treatment of human brain tumors. Structural and functional changes spread from the treatment surface and from the center to the periphery to involve the entire cerebral cortex exposed under a 5 mm craniectomy. The sequential changes spreading from the surface to the deepest cortical layer involve first astrocytes (1 hour), then endothelial cells and, ultimately, neurons. Thrombi were first noted in the microvasculature after 18 hours and coagulation necrosis of the entire area at risk occurred only after 48 hours. The results suggest that the photosensitizing agent crosses the intact blood-brain barrier and enters the astrocytic compartment where it becomes cytotoxic upon light activation. A comparison between the focal brain lesions of photodynamic therapy and those induced by middle cerebral artery occlusion suggests that cell damage evolves along different paths in these two forms of brain injury.

Animals↗