Natural (Mg,Fe)SiO3-ilmenite and -perovskite in the Tenham meteorite.

The minerals (Mg,Fe)SiO3-ilmenite and -perovskite were identified in the shock-induced veins in the Tenham chondritic meteorite. Both phases are inferred to have transformed from pyroxene at high pressures and temperatures by shock metamorphism. Columnar-shaped ilmenite grains, one of two types of morphologies, have a topotaxial relationship with neighboring pyroxene grains, indicating shear transformation. Granular-shaped perovskite grains showed a diffraction pattern consistent with orthorhombic perovskite, but these grains were not stable under the electron beam irradiation and became amorphous. The higher iron concentration in both phases compared with those experimentally reported may suggest their metastable transition from enstatite because of shock compression.

BRCA1 positively regulates FOXO3 expression by restricting FOXO3 gene methylation and epigenetic silencing through targeting EZH2 in breast cancer.

BRCA1 mutation or depletion correlates with basal-like phenotype and poor prognosis in breast cancer but the underlying reason remains elusive. RNA and protein analysis of a panel of breast cancer cell lines revealed that BRCA1 deficiency is associated with downregulation of the expression of the pleiotropic tumour suppressor FOXO3. Knockdown of BRCA1 by small interfering RNA (siRNA) resulted in downregulation of FOXO3 expression in the BRCA1-competent MCF-7, whereas expression of BRCA1 restored FOXO3 expression in BRCA1-defective HCC70 and MDA-MB-468 cells, suggesting a role of BRCA1 in the control of FOXO3 expression. Treatment of HCC70 and MDA-MB-468 cells with either the DNA methylation inhibitor 5-aza-2'-deoxycitydine, the N-methyltransferase enhancer of zeste homologue 2 (EZH2) inhibitor GSK126 or EZH2 siRNA induced FOXO3 mRNA and protein expression, but had no effect on the BRCA1-competent MCF-7 cells. Chromatin immunoprecipitation (ChIP) analysis demonstrated that BRCA1, EZH2, DNMT1/3a/b and histone H3 lysine 27 trimethylation (H3K27me3) are recruited to the endogenous FOXO3 promoter, further advocating that these proteins interact to modulate FOXO3 methylation and expression. In addition, ChIP results also revealed that BRCA1 depletion promoted the recruitment of the DNA methyltransferases DNMT1/3a/3b and the enrichment of the EZH2-mediated transcriptional repressive epigenetic marks H3K27me3 on the FOXO3 promoter. Methylated DNA immunoprecipitation assays also confirmed increased CpG methylation of the FOXO3 gene on BRCA1 depletion. Analysis of the global gene methylation profiles of a cohort of 33 familial breast tumours revealed that FOXO3 promoter methylation is significantly associated with BRCA1 mutation. Furthermore, immunohistochemistry further suggested that FOXO3 expression was significantly associated with BRCA1 status in EZH2-positive breast cancer. Consistently, high FOXO3 and EZH2 mRNA levels were significantly associated with good and poor prognosis in breast cancer, respectively. Together, these data suggest that BRCA1 can prevent and reverse FOXO3 suppression via inhibiting EZH2 and, consequently, its ability to recruit the transcriptional repressive H3K27me3 histone marks and the DNA methylases DNMT1/3a/3b, to induce DNA methylation and gene silencing on the FOXO3 promoter.

Cholinergic modulation of stellate cells in the mammalian ventral cochlear nucleus.

The main source of excitation to the ventral cochlear nucleus (VCN) is from glutamatergic auditory nerve afferents, but the VCN is also innervated by two groups of cholinergic efferents from the ventral nucleus of the trapezoid body. One arises from collaterals of medial olivocochlear efferents, and the other arises from neurons that project solely to the VCN. This study examines the action of cholinergic inputs on stellate cells in the VCN. T stellate cells, which form one of the ascending auditory pathways to the inferior colliculus, and D stellate cells, which inhibit T stellate cells, are distinguished electrophysiologically. Whole-cell recordings from stellate cells in slices of the VCN of mice demonstrate that most T stellate cells are excited by cholinergic agonists through three types of receptors, whereas all D stellate cells tested were insensitive to cholinergic agonists. Nicotinic excitation in T stellate cells has two components. The faster component was blocked by alpha-bungarotoxin and methyllycaconitine, suggesting that receptors contained alpha7 subunits; the slower component was insensitive to both. Muscarinic receptors excite T stellate cells by blocking a voltage-insensitive, "leak" potassium conductance. Our results suggest that cholinergic efferent innervation enhances excitation by sounds of T stellate cells, opposing the inhibitory action of cholinergic innervation in the cochlea that is conveyed indirectly through the glutamatergic afferents. The inhibitory action of D stellate cells on their targets is probably not affected by cholinergic inputs. Excitation of T stellate cells by cholinergic efferents would be expected to enhance the encoding of spectral peaks in noise.

FOXA1 repression is associated with loss of BRCA1 and increased promoter methylation and chromatin silencing in breast cancer.

FOXA1 expression correlates with the breast cancer luminal subtype and patient survival. RNA and protein analysis of a panel of breast cancer cell lines revealed that BRCA1 deficiency is associated with the downregulation of FOXA1 expression. Knockdown of BRCA1 resulted in the downregulation of FOXA1 expression and enhancement of FOXA1 promoter methylation in MCF-7 breast cancer cells, whereas the reconstitution of BRCA1 in Brca1-deficent mouse mammary epithelial cells (MMECs) promoted Foxa1 expression and methylation. These data suggest that BRCA1 suppresses FOXA1 hypermethylation and silencing. Consistently, the treatment of MMECs with the DNA methylation inhibitor 5-aza-2'-deoxycitydine induced Foxa1 mRNA expression. Furthermore, treatment with GSK126, an inhibitor of EZH2 methyltransferase activity, induced FOXA1 expression in BRCA1-deficient but not in BRCA1-reconstituted MMECs. Likewise, the depletion of EZH2 by small interfering RNA enhanced FOXA1 mRNA expression. Chromatin immunoprecipitation (ChIP) analysis demonstrated that BRCA1, EZH2, DNA methyltransferases (DNMT)1/3a/3b and H3K27me3 are recruited to the endogenous FOXA1 promoter, further supporting the hypothesis that these proteins interact to modulate FOXA1 methylation and repression. Further co-immunoprecipitation and ChIP analysis showed that both BRCA1 and DNMT3b form complexes with EZH2 but not with each other, consistent with the notion that BRCA1 binds to EZH2 and negatively regulates its methyltransferase activity. We also found that EZH2 promotes and BRCA1 impairs the deposit of the gene silencing histone mark H3K27me3 on the FOXA1 promoter. These associations were validated in a familial breast cancer patient cohort. Integrated analysis of the global gene methylation and expression profiles of a set of 33 familial breast tumours revealed that FOXA1 promoter methylation is inversely correlated with the transcriptional expression of FOXA1 and that BRCA1 mutation breast cancer is significantly associated with FOXA1 methylation and downregulation of FOXA1 expression, providing physiological evidence to our findings that FOXA1 expression is regulated by methylation and chromatin silencing and that BRCA1 maintains FOXA1 expression through suppressing FOXA1 gene methylation in breast cancer.

Assembly and disassembly of the peripheral architecture of the plant cell nucleus during mitosis

The architecture of the nuclei of higher plants includes a structure similar to the nuclear lamina of vertebrates. Changes in this structure were monitored during mitosis in carrot (Daucus carota L.) and celery (Apium graveolens L.) cells by immunofluorescence microscopy using an antibody that recognized the nuclear-matrix protein NMCP1. This protein has been shown to be localized exclusively at the periphery of the nucleus (K. Masuda et al. 1997, Exp Cell Res 232: 173-187). Immunofluorescence was recognized throughout cells in mitotic metaphase, although it was distributed predominantly in the mitotic spindle zone. At late anaphase or telophase, the immunofluorescence was localized around each set of daughter chromosomes. Immunofluorescence in newly formed daughter nuclei was restricted to the periphery of nuclei. This behavior was very similar to that of the nuclear lamina of vertebrates, suggesting that the structure located between the nuclear envelope and the chromosomes in plants disassembles and assembles in parallel with the disintegration and re-formation of the nuclear envelope.

Validity of 18F-fluorodeoxyglucose imaging with a dual-head coincidence gamma camera for detection of myocardial viability.

UNLABELLED: This study investigated the validity of myocardial 18F-fluorodeoxyglucose (FDG) imaging with a dual-head gamma camera operated in coincidence detection mode (DCD-I) by comparing this technique with conventional PET and SPECT with ultra-high-energy general-purpose collimators (UHGPs). METHODS: The subjects included 5 healthy volunteers and 20 patients with a history of myocardial infarction. FDG (370 MBq) was injected intravenously after 75-g oral glucose loading, and PET, UHGP SPECT and DCD-I were performed 45, 60 and 210 min, respectively, after the injection. The target-to-background ratio of each imaging method was evaluated for the healthy volunteers by comparing myocardial uptake with uptake in the upper lungs or left ventricular cavity. Agreement between the results of the various imaging methods was investigated for the myocardial infarction patients, as was the validity of DCD-I for assessing myocardial viability as judged by comparison with myocardial perfusion SPECT. The left ventricular wall was divided into 18 regions, and uptake was evaluated using a five-grade defect score (0 = normal; 1-3 = low uptake; 4 = defect). RESULTS: The mean ratio of myocardial counts to lung counts was lower on the DCD images (2.77 +/- 1.12) than on the UHGP SPECT images (3.69 +/- 0.98) (P < 0.05). In contrast, the mean ratio of myocardial counts to left ventricular cavity counts was higher on the DCD images (2.76 +/- 1.36) than on the UHGP SPECT images (1.98 +/- 0.70) (P < 0.05). For the patients, only 30.6% of the defect scores obtained by DCD-I agreed with the scores obtained by PET, and the defect scores in the inferior and septal walls were higher for the DCD images than for the PET images. When DCD-I was compared with PET without attenuation correction (AC), agreement improved to 58.3%. When corrected by a modified AC method, DCD-I improved to 48.1%. Agreement between UHGP SPECT and PET was 55.0%. Of the segments (64) for which the defect score of the myocardial perfusion image was greater than that for the FDG PET image, DCD-I without AC, DCD-I with AC and UHGP SPECT allowed an accurate diagnosis in 12 (18.8%), 31 (48.4%) and 43 (67.2%), respectively. CONCLUSION: The image quality of DCD-I is superior to that of UHGP SPECT. However, because the effect of attenuation is marked, accurate AC, by the transmission method, for example, is required to equal the validity of PET.

Corticotropin-releasing factor (CRF)-like immunoreactivity in the adrenal medulla.

Bovine adrenal medulla extract prepared by acid-acetone or acid methanol extraction showed two peaks of CRF-like immunoreactivity on Sephadex G-50 chromatography. One eluted near the void volume and another (low molecular weight CRF-like immunoreactivity) eluted slightly before arginine vasopressin (AVP), while most of the immunoreactivity in bovine hypothalamus coeluted with synthetic ovine CRF. When low molecular weight CRF fractions were chromatographed by reversed phase high performance liquid chromatography, three CRF-like immunoreactive peaks appeared. The first peak appeared near TRH, the second one eluted near AVP and the last one eluted near somatostatin. These three peaks of immunoreactivity showed ACTH releasing bioactivity in rat pituitary cells cultures. Therefore, the adrenal medulla-CRF-like substances might be tissue-CRF which may play a role to stimulate ACTH release in the severe stress conditions.

A new pulse sequence for "fast recovery" fast-scan NMR imaging.

This paper describes the "fast recovery" (FR) method for fast NMR imaging. The FR method combines a sequence of four RF pulses-alternating selective 90 degrees nutation pulses and nonselective 180 degrees pulses-with a gradient field pulse sequence which includes "spoiler" pulses to destroy the coherence between successive sequence cycles. We use the 2-D backprojection method of image reconstruction, but other imaging methods could be applied. The paper analyzes the behavior of the macroscopic magnetization-compares the FR method with other methods and proposes "figure of merit" expressions for relative signal-to-noise (S/N) ratios, scan time reduction ratios, and image contrast-and presents experimental results, including backprojection image reconstruction 2-D images and computed T1 and T2 images. For the FR method, in theory and practice, we find that, after each scan sequence cycle, magnetization is restored to equilibrium quickly and exactly; scan time can consequently be less than a tenth that for the saturation recovery method without any penalty in signal-to-noise ratio. Image contrast is even higher than that of the SR method, and compromise "optimum" sequence (interpulse timing) parameters give high image contrast for a wide range of tissue T1 and T2 (spin-lattice and spin-spin relaxation time) values.

Expression, cloning, and immunological analysis of buckwheat (Fagopyrum esculentum Moench) seed storage proteins.

cDNA of buckwheat (Fagopyrum esculentum Moench) was isolated from immature seeds harvested 14 days after pollination. Two genes, designated FA02 and FA18, were found to encode legumin-like proteins and were expressed during seed development. The deduced amino acid sequence of FA02 was identical to the N-terminal amino acid domain of BW24KD, which was believed to be a major buckwheat allergen (Urisu, A.; Kondo, Y.; Morita, Y.; Yagi, E.; Tsuruta, M.; Yasaki, T.; Yamada, K.; Kuzuya, H.; Suzuki, M.; Titani, K.; Kurosawa, K. Isolation and characterization of a major allergen in buckwheat seeds. In Current Advances in Buckwheat Research; Shinshu University Press: Matsumoto, Japan, 1995; pp 965--974). It was predicted that FA02 would be cleaved to generate two separate components, a 41.3 kDa alpha-subunit and a 21 kDa beta-subunit. Antiserum was raised against the deduced FA02 beta-subunit, and immunoblotting of total protein from buckwheat seeds (F. esculentum M. and Fagopyrum tartaricum Gaertn.) revealed that several groups of proteins reacted with the antiserum. Polypeptides in the 23--25 kDa range displayed the greatest reactivity.

Effects of iterative reconstruction on image contrast and lesion detection in gamma camera coincidence imaging in lung and breast cancers.

To investigate the effects of iterative reconstruction in 18F-fluorodeoxyglucose (FDG) gamma camera coincidence imaging (GCI), image contrast and visual detection obtained by using the iterative ordered-subsets expectation maximization (OSEM) reconstruction, in a phantom and in patients with lung cancer and breast cancer, were compared with those obtained by using the conventional filtered backprojection (FBP) reconstruction. Images of a cylindrical phantom containing hot spheres of various sizes (10-38 mm) were acquired by positron emission tomography (PET) and GCI at various sphere-to-background activity ratios. Forty-one consecutive patients with biopsy-proven cancer of lung (n = 20) and breast (n = 21) underwent PET and GCI on the same day after intravenous injection of 370 MBq of FDG. GCI images reconstructed by the OSEM and the FBP were compared. FDG PET was considered as the standard of reference. In GCI phantom images, OSEM yielded better contrast and signal-to-noise ratio (SNR) than FBP over the range of sphere sizes. Attenuation correction improved both the image measures and sphere detection obtained by the OSEM in GCI. In the study involving patients, FDG PET depicted 41 primary tumours and 25 metastatic lymph nodes. All of the tumours >2 cm in diameter (n = 25), six of the nine tumours 1.5-2.0 cm in diameter (67%), two of seven tumours <1.5 cm in diameter (29%), and 20 metastatic lymph nodes (80%) were detected in attenuation uncorrected GCI reconstructed by the OSEM as well as the FBP. The undetected lesions in GCI were identical between the OSEM and the FBP reconstructions. OSEM yielded significantly greater tumour-to-background (T/B) ratios and lower noise than FBP in GCI (T/B ratios, 4.1+/-3.2 vs 3.7+/-2.7, P = 0.02; noise, 0.09+/-0.04 vs 0.14+/-0.05, P<0.0001). In conclusion, OSEM yielded better image contrast and less noise than the FBP in GCI, but the lesion detection obtained by the OSEM and the FBP in attenuation uncorrected GCI in patients with lung cancer and breast cancer were similar. Phantom data suggest the potential of OSEM for improving lesion detection in GCI after attenuation correction.

The nature of ventricular arrhythmias during ergonovine-induced vasospastic angina pectoris.

The peak incidence of ventricular fibrillation in acute myocardial infarction usually occurs during the first hours after the onset. Electrophysiological changes immediately after the onset have been studied in animal models, but are still incompletely understood in humans. For clarification of the characteristic features of ventricular arrhythmias during acute myocardial ischemia, ventricular arrhythmias were studied in 81 patients with vasospastic angina pectoris induced by ergonovine. Ventricular arrhythmias occurred in 45 of these patients, including ventricular tachycardia in 15, and ventricular fibrillation requiring repeated DC defibrillation in two patients. Most ventricular extrasystoles occurred before the ST segment reached maximum elevation, while reperfusion arrhythmias were less common. In many patients the coupling intervals varied, and the configuration was multiform. It is concluded that ventricular arrhythmias occurring during ergonovine-induced coronary spasm show different characteristics from those occurring during chronic ischemia. As the arrhythmias in this study seem, in some ways, to resemble arrhythmias occurring at the onset of myocardial infarction, the results might provide useful information on ventricular arrhythmias in myocardial ischemia in humans.

Vector U loop in patients with old myocardial infarction.

The U loop of the vectorcardiogram was examined qualitatively and quantitatively in 100 normal subjects and 67 patients with old myocardial infarction, using a direct-writing vectorcardiograph with memory function. In the control group, the U loop was directed to the left, anteriorly and inferiorly, and it was inscribed counterclockwise in the horizontal plane. In patients with anterior myocardial infarction, the U loop tended to be displaced to the right, and in patients with inferior myocardial infarction to the right and superiorly. The shape of the U loop in patients was also different from that of normal subjects. The maximum U vector was significantly smaller in magnitude both in patients with anterior and inferior myocardial infarction than that of normal subjects (p less than 0.01). In patients with ventricular aneurysm, the magnitude of the maximum U vector was significantly smaller and its direction was displaced more to the right and posteriorly than those without aneurysm (p less than 0.01). In standard 12-lead electrocardiogram (ECG), observation of the U wave in patients with old myocardial infarction was difficult, especially in the limb lead, because of the small size of the U wave. Therefore, vectorcardiographic observation may be more useful than electrocardiographic observation for the analysis of the U wave in patients with old myocardial infarction.

Paraganglioma associated with hypoglycemia.

A 63-year-old female had frequent attacks of palpitations with sweating and occasional loss of consciousness for 7 years before admission. These symptoms and documented hypoglycemia (blood glucose: 46 mg/dl) suggested insulinoma, but computed tomography demonstrated a retroperitoneal tumor which was diagnosed as a paraganglioma by histological examination of the resected specimen.

Efficacy of a continuous syringe extraction method for monitoring hemodialysis ultrafiltrate.

The evaluation of hemodialysis ultrafiltrate is essential for the assessment of uremic toxins, dialyzer net performance, protein catabolic rate, and safety and environmental protection. Total dialysate collection (TDC), however, is technically far from the daily procedure used. In the present study, use of a continuous syringe extraction method (CSEM) as a substitute for TDC was tested to determine its comparative effectiveness. Measurements of urea nitrogen, creatinine, phosphate, beta2-microglobulin, and albumin were simultaneously obtained by both TDC and CSEM in 20 dialysis sessions. CSEM showed an extremely significant correlation with TDC for these values. The correlation coefficients were >0.97 for these indicators and the value of Fisher's r to z were all <0.001. Taken together, these data indicate that CSEM is an effective substitute for TDC. With use of CSEM, the evaluation of spent dialysate could become as a daily procedure.

Brain catecholamines in spontaneously hypertensive and DOCA-salt hypertensive rats.

The concentrations and alpha-methyl-p-tyrosine (alpha-MPT) induced disappearance of catecholamines, adrenaline, noradrenaline and dopamine, were measured in selected areas of the brainstem and hypothalamus of spontaneously hypertensive rats (SHR) and deoxycorticosterone acetate (DOCA)-salt hypertensive rats. The catecholamine levels were measured by a sensitive radioenzymatic assay method combined with microdissection of the rat brain. The adrenaline concentration was higher in the area A1 of young SHR, but not in adult SHR, than in age-matched control rats. Noradrenaline concentrations and the alpha-MPT induced noradrenaline disappearance were less in the rostral part of the nucleus tractus solitarii (NTS) and the nucleus hypothalamic anterior of young SHR, and in the rostral part of the NTS of adult SHR. On the other hand in DOCA-salt hypertensive rats, the concentrations of adrenaline and noradrenaline were the same as in control rats in the examined areas. The alpha-MPT induced noradrenaline disappearance was less in the rostral part of the NTS of DOCA-salt hypertensive rats. Dopamine concentrations and the alpha-MPT induced dopamine disappearance were the same in the examined areas of SHR and DOCA-salt hypertensive rats. The results suggest that SHR have a change in adrenergic neural activity in the brainstem and a decrease in noradrenergic neural activity in the brainstem and hypothalamus while DOCA-salt hypertensive rats have a decrease in noradrenergic neural activity in the brainstem. Such changes in brain catecholaminergic neurons may have played an important role in the development of hypertension in these rats.

Efferent vestibular fibers to otolith organs in guinea pigs.

The origins of efferent vestibular fibers to the otolith organs in guinea pigs, examined with the use of retrograde transport of HRP were: i) the bilateral lateral portions of the facial genu and ii) the ipsilateral reticular nucleus at the level of the facial nucleus. These origins are almost the same as those of the efferent fibers to the three semicircular canals which we have previously identified in cats; however, they differ in certain respects.

A primary hepatic carcinoid tumor: evaluation by computed tomography and magnetic resonance imaging.

A case of primary hepatic carcinoid tumor is reported. A 40-year-old woman was admitted to our hospital because of an epigastric mass. Ultrasonography demonstrated a heterogeneous, hyperechoic mass with hypoechoic area in the left lobe of the liver. CT showed a hypodense lesion with more hypodense areas. Enhanced CT showed slight retention of contrast medium and a low density area that suggested necrosis in the tumor. The mass appeared as a heterogeneous area of low signal intensity with sharp margin on T1-weighted MRI. T2-weighted MRI revealed a high intensity mass with higher intensity foci. Left lateral segmental hepatic resection was performed. Immunohistochemical and electron microscopic studies revealed the carcinoid tumor. Postoperatively, careful, intensive search revealed no other primary source of the tumor. The higher intensity foci were microscopically identified as intratumoral hemorrhage, corresponding to hemorrhagic areas on the cut surface. Therefore, T2-weighted MRI might reflect the pathological characteristics of the tumor.

Penetrating injury of the vertebral artery caused by a nail gun: preoperative embolization.

We describe a penetrating injury of the vertebral artery caused by a nail gun. The artery was preoperatively embolized using mechanical detachable coils (MDC).

Surgical difficulties and postoperative problems associated with cochlear implants.

Since 1987 we have implanted 60 severely hearing-impaired adults with the Nucleus Minisystem-22 cochlear implant. In most cases, the surgical procedures were performed with satisfactory results and very few postoperative problems. The complications associated with cochlear implant surgery include difficulty in drilling the ossified cochlea, electrode displacement, and insufficient insertion of the electrode. Postoperative problems include dizziness, tinnitus, facial nerve spasm, receiver-stimulator migration or extrusion, infection of the scalp flap, and facial nerve palsy. In most cases, these complaints did not develop into serious conditions.

Relationship between morbidity and body mass index of mariners in the Japan Maritime Self-Defense Force Fleet Escort Force.

To establish a practical weight management program for mariners in the Japan Maritime Self-Defense Force (JMSDF) Fleet Escort Force, the relationship between morbidity and body mass index (BMI) was studied. To estimate morbidity, 10 medical problems were used as indices (hyperlipidemia, hyperuricemia, diabetes mellitus, lung disease, heart disease, upper gastrointestinal tract disease, hypertension, renal disease, liver disease, and anemia). A curvilinear relationship was found between morbidity and BMI, in which a BMI of 17.5 was associated with the lowest morbidity. This curvilinear pattern was more complex than a curve reported previously for Japanese civilians. Using the present curve and aiming for a BMI of 17.5 will help in the design and implementation of a practical management program for health promotion in the JMSDF.