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Biomedical subjects

M Yamane

Publications and source records attributed to M Yamane.

At least 73 records · Page 4Linked to original sources

High-performance liquid chromatograpy-thermospray mass spectrometry of omega-hydroxy polyunsaturated fatty acids from rat brain homogenate.

A method for the analysis of omega-hydroxy polyunsaturated fatty acids (omega-HPUFAs) in rat tissue homogenate, supplemented with NADPH and homo-gamma-linolenic acid [20:3(n-6)], arachidonic acid [20:4(n-6)], eicosapentaenoic acid [20:5(n-3)] or docosahexaenoic acid [22:6(n-3)] as a substrate was developed. By ion analysis of chromatograms obtained with reversed-phase HPLC-thermospray MS, many omega-HPUFAs corresponding to each precursor fatty acid could be characterized by the high intensity of the molecular ion (MH+) and quasimolecular ion (MNH4+, MNa+), while other common HPUFAs were characterized by the high intensity of the base ion of MH+--H2O. On a selected-ion monitoring chromatogram of rat brain homogenate, significant amounts of omega-HPUFA from each precursor fatty acid, especially from 22:6(n-3), were detected compared with the amounts found in rat large intestine homogenate. Based on these results, a highly active NADPH-dependent omega-oxidation system is suggested for rat brain homogenate resulting in extensive oxidation of 22:6(n-3).

Animals↗

High-performance liquid chromatography-thermospray mass spectrometry of epoxy polyunsaturated fatty acids and epoxyhydroxy polyunsaturated fatty acids from an incubation mixture of rat tissue homogenate.

A method for the analysis of epoxy polyunsaturated fatty acids (EpPUFAs) and epoxyhydroxy polyunsaturated fatty acids (EpHPUFAs) in rat tissue homogenate, with homo-gamma-linolenic acid (20:3, n - 6), arachidonic acid (20:4, n - 6), eicosapentaenoic acid (20:5, n - 3) or docosahexaenoic acid (22:6, n - 3) as a substrate, has been developed. Extraction with dichloromethane at pH 4-5 and concentration in the presence of pyridine were performed. Spectral analysis of chromatograms obtained with high-performance liquid chromatography-thermospray mass spectrometry showed the presence of EpPUFAs, EpHPUFAs and dihydroxy metabolites (DiHPUFAs) of EpPUFAs corresponding to each precursor fatty acid. On a selected-ion monitoring chromatogram, many EpPUFAs, EpHPUFAs and DiHPUFAs in an extract from an incubation mixture of each precursor fatty acid in aged rat tissue homogenate were detected simultaneously within 70 min. EpPUFAs and DiHPUFAs derived from 20:3 (n - 6) or 20:5 (n - 3) were detected in significant amounts. From these results, a highly active cytochrome P450 system or non-enzymic oxidative reactions in aged rat tissue homogenate were suggested.

Animals↗

Analysis of lower aliphatic aldehydes in water by micellar electrokinetic chromatography with derivatization to 2,4-dinitrophenylhydrazones.

The analysis of lower aliphatic aldehydes in water following derivatization to 2,4-dinitrophenylhydrazones, was investigated by micellar electrokinetic chromatography (MEKC). The effects of the buffer pH and acetonitrile addition on the separation were investigated. Under optimized conditions the 2,4-dinitrophenylhydrazine (DNPH) reagent was completely separated from four DNPH derivatized aldehydes with high separation efficiency. DNPH-formaldehyde and DNPH-acetaldehyde were also completely separated from one another. Their relative standard deviation of migration times and peak areas were within 1.0% and 3.3%, respectively. The reproducibility of peak areas in this MEKC analysis was as good as that of high-performance liquid chromatography (HPLC) analysis with the same derivatization method. The recovery values from tap and river water were in the range of 97-102%. The detection limits of formaldehyde and acetaldehyde were 0.05 mg/L and 0.08 mg/L respectively.

Acetaldehyde↗

Increased plasma cholesteryl ester transfer protein in obese subjects. A possible mechanism for the reduction of serum HDL cholesterol levels in obesity.

It is well known that obesity is frequently associated with low levels of serum high-density lipoprotein (HDL) cholesterol. However, the mechanism for this reduction has not been fully clarified. Cholesteryl ester transfer protein (CETP) transfers cholesteryl ester from HDL to apolipoprotein B-containing lipoproteins and plays an important role in regulating the concentration and composition of HDL. To elucidate the mechanism for the reduction of serum HDL cholesterol in obesity, we analyzed serum lipoproteins, CETP, and postheparin lipoprotein lipase (LPL) and hepatic triglyceride lipase (HTGL) activities in 30 obese subjects (17 women and 13 men, age 44 +/- 14 years, mean +/- SD). We also investigated the relationship between these variables, total adiposity, and indices of body fat distribution. The average body mass index of the obese subjects was 33.1 +/- 4.8 kg/m2 (range, 26.4 to 43.8 kg/m2). The obese subjects showed significantly lower serum HDL cholesterol levels than control subjects (1.04 +/- 0.28 versus 1.50 +/- 0.34 mmol/L, P < .01). In the obese subjects, both activities and protein mass of CETP and postheparin HTGL activities were significantly increased, whereas postheparin LPL activities were significantly decreased. CETP activities, independent of postheparin HTGL and LPL activities, were correlated negatively with HDL cholesterol (r = -.39, P < .05) and the cholesteryl ester to triglyceride ratio of HDL2 and HDL3 (r = -.36, P < .05; r = -.46, P < .05, respectively).(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

[Surgical treatment of atherosclerotic vascular diseases associated with coronary artery disease].

Selective coronary angiography to determine the incidence of coronary artery disease (CAD) has been performed in the 38 patients with atherosclerotic vascular disease. significant CAD was demonstrated in 17 of 23 (73.8%) patients with angina pectoris or previous myocardial infarction, in 4/15 (26.7%) patients who had no clinical evidence of CAD. There were no early and late postoperative death in the 9 patients who underwent simultaneous coronary artery bypass graft and vascular operations or percutaneous coronary angioplasty, on the other hand, there were one early and four late death in the 11 patients who underwent only vascular procedure.

Aged↗

Excitation and inhibition of neuronal activity in the pontine micturition center by pelvic rectal and pudendal anal afferents in dogs.

To examine whether or not the pontine micturition center (PMC) is involved in the inhibition of the micturition reflex by pelvic rectal and pudendal anal afferents, neuronal activity in the PMC was observed during inhibition of this reflex in paralyzed decerebrate dogs. Discharge of pelvic vesical branches (VBs) waxed and waned at a rhythm of about 2 Hz during the micturition reflex, which was activated by continuous stimulation of the contralateral VBs. This rhythmic discharge was modulated by continuous stimulation of contralateral pelvic rectal branches (RBs) superimposed on the VB stimulation. The modulation was composed of three effects; initial inhibition, augmentation and late inhibition. However, not all of the three effects were obvious in some dogs. One-sixth of 118 neurons examined in the pontine area ventromedial to the locus ceruleus exhibited rhythmic burst firings which preceded the rhythmic discharge of VBs by about 150 ms. Therefore, these pontine neurons are assumed to be output neurons of the PMC. The rhythmic firings of pontine neurons were augmented during continuous RB stimulation independent of the inhibitory and/or augmentative effects of the RB stimulation on the reflex discharge of the VBs. In contrast, the rhythmic firings of the pontine neurons and the reflex discharge of VBs were inhibited by mechanical stimulation of the anal canal and perineal hairs. These results suggest that the PMC is involved in the inhibition of the micturition reflex produced via pudendal afferents but not in that produced by pelvic rectal afferents, and that pelvic and pudendal afferents project to the PMC through separate pathways.

Animals↗

Neuro-otological findings in patients with spinocerebellar degeneration: with special regard to the duration of illness.

Neuro-otological findings in 150 patients with spinocerebellar degeneration were analyzed based on the duration of illness since onset. The results of each kind of neuro-otological examination varied in close relation to the duration of illness. Also, the results of each kind of examination correlated closely. In particular, the optokinetic nystagmus test and the visual suppression test correlated most closely. Neuro-otological examinations were confirmed to be of great value in gauging progression in spinocerebellar degeneration.

Adolescent↗

Role of hepatic vitamin A and lipocyte distribution in experimental hepatic fibrosis.

Lipocytes are the major site of hepatic vitamin A storage, and they have been demonstrated to lose their vitamin A content in the process of hepatic fibrosis. To investigate the relationship between hepatic vitamin A content and the degree of hepatic fibrosis, we measured levels of retinyl palmitate and retinol in the CCl4-induced fibrotic liver using high-performance liquid chromatography. We estimated hepatic collagen content using a spectrophotometric analysis with sirius red, and also by measuring hydroxyproline levels. Lipocytes were detected by an immunoperoxidase method with anti-desmin antibody, and were counted morphometrically through a Texture Analyzing System. A significant negative correlation was observed between the level of retinyl palmitate and collagen content (r = -0.64) as well as the hydroxyproline level (r = -0.69) in the CCl4-induced fibrotic liver. In the process of fibrosis, hepatic retinol levels were elevated in association with a decrease in retinyl palmitate. In particular in the early stage of fibrosis, lipocytes increased remarkably in number in fibrotic areas in spite of a decrease in total hepatic vitamin A. The present study suggests that an increase in hepatic retinol as well as a decrease in retinyl palmitate may facilitate the process of hepatic fibrosis produced by lipocytes.

Animals↗

Disseminated pancreatic polypeptidioma.

Pancreatic polypeptidioma, a pancreatic endocrine tumor, is an extremely uncommon disease and its clinical features and responses to therapy are not well known. We present a 33-year-old woman with disseminated pancreatic polypeptidioma, who subsequently showed various signs and symptoms of metastases, including bone pain, cranial nerve palsy, spinal block, and hematuria, and died 22 months after the presentation. Responses to various therapeutic regimens including hepatic arterial embolization, radiation therapy, systemic chemotherapy, and administration of interferon-alpha or somatostatin analogue, are discussed. Particular note in this case is a prompt response of bone metastases to the radiotherapy.

Adult↗

High-performance liquid chromatography-thermospray mass spectrometry of hydroperoxy polyunsaturated fatty acid acetyl derivatives.

A method for the analysis of hydroperoxy polyunsaturated fatty acids was developed. The hydroperoxy groups were acetylated by acetic anhydride, and the mixture was partially purified on a Sep-Pak C18 cartridge and analysed by high-performance liquid chromatography with thermospray mass spectrometry. Generally, the base ion, [M+H - n(60)]+ or [M+H - n(60) - n(H2O)]+, is produced through elimination of acetic acid or water (n = number of hydroperoxy groups). The detection limit for these derivatives was ca. 1 pmol at concentrations of hydroperoxy polyenoic acids prior to derivatization. Using this method, many hydroxy and hydroperoxy polyunsaturated fatty acid derivatives could be detected simultaneously within 30 min on a selected-ion monitoring detection chromatogram without a gradient system. The assay was successfully applied to hydroxy and hydroperoxy polyunsaturated fatty acids from an incubation mixture of rat brain homogenate to which polyunsaturated fatty acids had been added.

Acetylation↗

High-performance liquid chromatography-thermospray mass spectrometry of hydroxy-polyunsaturated fatty acid acetyl derivatives.

A method for the determination of hydroxy-polyenoic acids has been developed. Hydroxy groups of hydroxy-polyunsaturated fatty acid were acetylated by acetic anhydride, and the mixture was partially purified on a Sep-Pak C18 cartridge and analysed by high-performance liquid chromatography combined with thermospray mass spectrometry. Using this method, fifteen kinds of hydroxy-polyunsaturated fatty acid derivative could be detected simultaneously within 30 min on a selected-ion monitoring detection chromatograph without a gradient system. Generally, the base ion, [M + H - n(60)]+, is produced through elimination of acetic acid (n = number of the hydroxy group of hydroxy-polyunsaturated fatty acid). The detection limit for these derivatives was ca. 0.2 pmol at the levels of hydroxy-polyenoic acids prior to derivatization. They could be analysed in the range 0.5-25 pmol. The assay was successfully applied to hydroxy-polyunsaturated fatty acids from an incubation mixture of rat brain homogenate to which polyunsaturated fatty acid was added.

Animals↗

Subzonal insemination of a single mouse spermatozoon with a personal computer-controlled micromanipulation system.

A personal computer-controlled micromanipulation system was developed for automatic injection of spermatozoa into the perivitelline space of mouse ova. A pair of three-dimensional hydraulic micromanipulators driven by pulse motors was used for this automatic system. The pulse signals that regulate the motors are initiated by the computer program, and these signals cause the micromanipulator to move the microtool precisely. The computer program was designed to perform the most effective movements of the sperm injection needle used during manual micromanipulation. Prior to the manipulation, the computer locates the tip of the injection needle and the end of the egg-holding pipette in the microscope field using image processing. The trajectory of the injection needle is determined according to these initial positions. Using this robotic system, subzonal insemination with a single mouse spermatozoon was attempted in a total of 143 ova. The sperm insertion was successfully completed in all cases without damaging any of the ova. Spermatozoa treated with ionophore A23187 and those without the treatment were used. The fertilization rate (68.8%) of the ova inseminated with treated sperm was significantly higher than that (37.5%) obtained with the nontreated sperm (P less than 0.05). These findings suggest the feasibility and potential for further applications of a robotic microinsemination system and, in addition, that a higher fertility rate in the subzonal insemination of mouse ova can be achieved with the ionophore treatment of spermatozoa.

Animals↗

Stapedial reflex in Parkinson's disease.

In 27 patients with Parkinson's disease (PD), stapedial reflexes were measured using impedance audiometry and compared with those of 11 age-matched control subjects. The reflex threshold of PD patients was lower than that of control subjects. A prolongation of contraction time (C50) and relaxation time (D50) was revealed. Between patients with and without L-dopa, there was no significant difference for any reflex parameter. But, the D50 of patients without anticholinergic drugs was longer than that of patients with anticholinergic drugs. The authors could not find any relationship between the severity of PD and the reflex parameters. The authors assume that the prolongation of reflex parameters might be attributed to the hyperactivity of the indirect pathways of the stapedial reflex.

Acoustic Impedance Tests↗

Beta(+)-thalassemia with hemochromatosis.

A 64-year-old man was admitted due to ascites. Laboratory data showed hemoglobin 6.7 g/dl, mean corpuscular volume 82 fl, and ferritin 2,360 ng/ml. Liver biopsy showed hemochromatosis. The diagnosis of beta-thalassemia was suggested by a decreased ratio of beta/alpha-globin synthesis in vitro (0.26). Cloning of the beta-globin gene showed A-to-G mutation in the first base of the ATA box. He was confirmed to be homozygous for this specific allele by beta-gene complex analysis and analysis of Southern blot hybridization of the alpha- and beta-globin genes. His two sons were confirmed to be heterozygous for this allele.

Adult↗

[Comparison of CT and MRI in diagnosis of parapharyngeal tumors].

Recently, the usefulness of CT and MRI in diagnosing parapharyngeal tumors has been established. At the same time, several modalities for these imagings, i.e. plane and enhanced CT, T1, 2 weighted MRI and Gadolinium enhanced MRI, have been developed. We compared the image findings of 12 tumors involving the parapharyngeal space with their operative records. T1 was most suited to diagnosing the sites of origins of tumors because of its superior depiction of the internal carotid artery and parapharyngeal fat. Enhanced CT was also superior in depicting these structures, but in cases where the parapharyngeal fat was diminished, or the parapharyngeal fat had been invaded by malignant tumor, enhanced CT was inferior to T1. The spatial resolution of T2 was poor, T2 was thus not useful in diagnosing the sites of origins of tumors. Because tumor intensity tended to be similar to that of fat in Gadolinium enhanced MRI imaging, Gadolinium was not as useful as T1. In diagnosing the extents of tumors, we examined the depiction of both tumor-fat and tumor-muscle interfaces. With regard to the depiction of the tumor-fat interface, enhanced CT and T1 were most useful. In diagnosing the tumor-muscle interface, Gadolinium enhanced MRI was most suitable. The net result is that MRI is superior to CT.

Adipose Tissue↗

High-performance liquid chromatography-thermospray mass spectrometry of prostaglandin and thromboxane acetyl derivatives.

A quantitative analysis of prostaglandin-related substances has been developed. Hydroxyl groups of prostaglandin and thromboxane were acetylated by acetic anhydride, the mixture was partially purified on a Sep-Pak C18 cartridge and analysed by high-performance liquid chromatography combined with thermospray mass spectrometry. Using this method, twenty kinds of prostaglandin derivative could be detected simultaneously within 11 min on a selected-ion monitoring detection chromatogram without a gradient system. Generally, the base ion, [M + H - n(60)]+, is produced through elimination of acetic acid (n = number of the hydroxyl group of prostaglandin or thromboxane). The detection limit for these derivatives was ca. 0.2 pmol at the levels of prostaglandin-related substances prior to derivatization. They could be analysed in the range 0.5-10 pmol. The assay was successfully applied to prostaglandin-related substances in human seminal fluid and rat brain.

Animals↗