Effects of isoflavone-rich red clover extract on blood glucose level: A randomized, double-blind, placebo-controlled trial.

High blood glucose is associated with increased risk of various diseases. Red clover (RC; Trifolium pratense L.) is an edible legume whose sprout is rich in isoflavones such as formononetin and biochanin A. We examined the effects of RC extract on postprandial and fasting blood glucose level, using a randomized, double-blind, placebo-controlled trial with 36 participants, aged 25 to 64 years, who were randomly assigned to receive either 1.91 g of RC extract (containing 8 mg formononetin and 1.8 mg biochanin A) or placebo. Each participant ingested the assigned test food daily for 8 weeks, and at the oral maltose tolerance test (OMTT). Initially, the two groups did not significantly differ in OMTT results. However, fasting insulin levels at 8 weeks were significantly lower in the RC group (4.76 µIU/ml at Week 0 to 4.01 µIU/ml at Week 8) with a significant interaction (P = 0.046). Subgroup analysis showed that change in blood glucose level (blood glucose ΔC) tended to decrease late in the trial period during OMTT in the ≤50-year-old RC group, as did fasting blood glucose and insulin levels at 8 weeks; hemoglobin A1c was also significantly reduced in this subgroup (5.36% at Week 0 to 5.28% at Week 8) with a significant interaction (P = 0.040). These results suggest that the daily intake of RC could reduce blood glucose, particularly for those ≤50 years old. Formononetin-an α-glucosidase inhibitor-is considered to be the major functional molecule for these effects. Therefore, intake of RC that contains formononetin might help blood glucose control.

Benzyl Isothiocyanate Produced by Garden Cress (Lepidium sativum) Prevents Accumulation of Hepatic Lipids.

We determined the physiological effects of glucotropaeolin-rich lyophilized garden cress sprout powder (GC) administered to fasting and nonfasting mice. High-performance liquid chromatography analysis revealed glucotropaeolin (57.4±1.1 mg/g dry weight) as a major phytochemical constituent of GC. Decreasing tendency in body weight and feeding efficiency ratio were detected in the group of mice fed 0.05% (w/w) GC (GC0.05). Nonfasting mice exhibited significantly lower liver weights that were unchanged after fasting. Decreased total lipid (TL) and triglyceride (TG) levels in the liver were detected in the nonfasted GC0.01 and GC0.05 groups, but only in TLs of the fasted groups. The levels of plasma TGs and nonesterified fatty acids of the GC0.05 group, which remained unchanged during nonfasting, decreased after fasting. To determine its effects on the accumulation of lipids in the liver, the glucotropaeolin aglycone, benzyl isothiocyanate (BITC), was added to the liver-derived HepG2 human cell line cultured in a medium containing a high concentration of D-glucose (4,500 mg/L D-glucose) (HG group) or 1 mM oleic acid (SO group). Toxicity was not detected when cells were treated with as much as 5 µM BITC; however, lipid accumulation was inhibited by BITC in a concentration-dependent manner in the HG groups. The same effect was observed when 2 µM BITC was added to the diet of the SO groups. These results suggest that moderate levels of GC or BITC are useful for reducing liver and plasma TGs.

Red Clover (Trifolium pratense L.) Sprout Prevents Metabolic Syndrome.

We examined the prevention effect of red clover (Trifolium pratense L.) sprout on metabolic syndrome using a high-carbohydrate and high-fat diet (Western diet; WD)-induced male C57BL/6J obese model mouse. Red clover sprout-lyophilized powder (RC) contained 3.5 mg/g dry-weight of formononetin as a major phenolic compound, as analyzed by high performance liquid chromatography. Supplementation of 0.3% (w/w) RC in a WD (WD+RC) showed an anti-obesity effect and ameliorated lipid metabolism in the obese model mice. Additionally, fasting plasma glucose levels were significantly reduced in the WD+RC group. Administration of 0.1 mg/kg formononetin reduced the postprandial blood glucose level, as assessed using the oral maltose tolerance test. However, no significant formononetin intake effect was observed on the plasma insulin level. These results suggest that the formononetin contained in red clover sprout inhibits α-glucosidase and thereby contributes to reducing the postprandial blood glucose response in mice.

Nucleophilic Arylation of Halopurines Facilitated by Brønsted Acid in Fluoroalcohol.

Various aryl-substituted purine derivatives were synthesized through the direct arylation of halopurines with aromatic compounds, facilitated by the combination of triflic acid and fluoroalcohol. This metal-free method is complementary to conventional coupling reactions using metal catalysts and reagents for the syntheses of aryl-substituted purine analogues.

Prism-pair interferometer for precise measurement of the refractive index of optical glass by using a spectrum lamp.

A prism-pair interferometer for a spectrum lamp was developed for precise measurement of the refractive index of a prism of optical glass. Previously we reported the prism-pair interferometer with a He-Ne laser light source, resulting in a measurement uncertainty of 1.1×10⁻6. However, most of the refractive-index values managed by optical glass manufacturers are conventionally measured with spectrum lamps. We have optimized the prism-pair interferometer for spectrum lamps and implemented a signal-processing technique from Fourier-transform spectroscopy. When the refractive index is measured, the wavelength of the spectrum lamp is simultaneously calibrated by part of the interferometer, so that the resulting refractive index is traceable to a national standard of length. The combined standard uncertainty for a refractive index measured with the e-line (546 nm) of a Hg lamp is 6.9×10⁻6.

Novel insights into the function of Arabidopsis R2R3-MYB transcription factors regulating aliphatic glucosinolate biosynthesis.

Arabidopsis transcription factors, MYB28, MYB29 and MYB76, positively regulate aliphatic glucosinolate (AGSL) biosynthesis. Mutual transcriptional regulation among these MYB genes makes it difficult to elucidate their individual function simply by analyzing knock-out mutants or ectopically overexpressing lines of these genes. In this study, we constructed transgenic lines expressing each MYB gene driven by its own promoter in the myb28myb29 background, where the expression of the endogenous MYB28, MYB29 and MYB76 was repressed with no AGSL accumulation. In leaves, transgenic MYB28 expression activated AGSL biosynthetic genes and restored accumulation of AGSLs with short side chains. Transgenic MYB29 expression activated the same biosynthetic pathway, but induction of the genes involved in side chain elongation was weaker than that by MYB28, resulting in a weaker recovery of AGSLs. Neither MYB28 nor MYB29 recovered long-chain AGSL accumulation. MYB76 was considered to require both MYB28 and MYB29 for its normal level of expression in leaves, and could not activate AGSL biosynthesis on its own. Interestingly, the accumulation in seeds of long- and short-chain AGSLs was restored by transgenic expression of MYB28 and MYB76, respectively. A sulfur stress experiment indicated that MYB28 expression was induced by sulfur deficiency, while the expression levels of MYB29 and MYB76 were positively correlated with sulfur concentration. This study illustrated how the individual MYBs work in regulating AGSL biosynthesis when expressed alone under normal transcriptional regulation.

Fully interferometric three-dimensional imaging spectrometry using hyperbolic-type volume interferogram.

A signal-processing method is proposed in the fully interferometric three-dimensional (3D) imaging spectrometry. This processing computes a 3D interferogram, in which recorded fringe patterns do not directly reflect wavefront forms propagated from a polychromatic light source under measurement. This paper presents a procedure for signal processing including a synthesis of the 3D interferogram and retrieval of a set of spectral components of 3D images. We demonstrate retrieving 3D images for spectral components of two planar light sources by means of the proposed method. The procedure to synthesize the 3D interferogram in this method suggests the possibility of direct measurement of the 3D interferogram.

[Anesthetic management of a patient with SAPHO syndrome: a case report].

We report a case of SAPHO syndrome accompanying progressive osteoarthritis. In this 43-year-old woman, difficult intubation was expected due to unilateral hyperostosis of the mandible and spinal degeneration. As far as we know, anesthetic management of this disease has not been reported. Therefore we made an anesthetic plan based on spinal degenerative disease which was considered to have similar problems of intubation and the case was managed without problems. As this syndrome is a progressive disorder, careful perioperative management is required each time.

Prism-pair interferometry by homodyne interferometers with a common light source for high-accuracy measurement of the absolute refractive index of glasses.

A prism-pair interferometer comprising two homodyne interferometers with a common light source was developed for high-precision measurements of the refractive index of optical glasses with an uncertainty of the order of 10(-6). The two interferometers measure changes in the optical path length in the glass sample and in air, respectively. Uncertainties in the absolute wavelength of the common light source are cancelled out by calculating a ratio between the results from the interferometers. Uncertainties in phase measurement are suppressed by a quadrature detection system. The combined standard uncertainty of the developed system is evaluated as 1.1×10(-6).

[Successful management of a patient with myotonic dystrophy under total intravenous anesthesia with propofol, remifentnil and rocuronium bromide, combined with epidural anesthesia].

In general anesthesia for a patient with dystrophia myotonica (DM), respiratory depression and muscle weakness by opioid, as well as prolongation of the effect of muscle relaxant are seen postoperatively. Therefore it is desirable to choose agents with short duration of action and to dose these medicines to the minimum. We report a case of a 45-year-old woman with DM who underwent laparotomy for uterine cancer under general anesthesia combined with epidural anesthesia. Epidural catheter was placed from T 11-12, and anesthesia was inducted with propofol and remifentanil (RF). We administered rocuronium bromide (RB) 5 mg while watching TOF ratio with a muscle relaxation monitor (TOF-Watch). T1 became 0 after giving a dose of 10 mg, and intubation was performed. We maintained anesthesia by propofol and RF combined with epidural anesthesia. TOF ratio was restored to around 80% 90 minutes after RB administration, but we did not give supplemental doses because the operation went well smoothly. Recovery was smooth and fast. The respiratory depression and the muscle spasm were not noticed. RB and RE both with short duration of action, are useful in anesthesia management in DM cases.

Widely targeted metabolomics and coexpression analysis as tools to identify genes involved in the side-chain elongation steps of aliphatic glucosinolate biosynthesis.

Amino acid and glucosinolate biosynthesis are two interdependent pathways; amino acid synthesis as a part of primary metabolism provides the precursors for glucosinolate biosynthesis in secondary metabolism. In our previous studies, the combination of coexpression analysis and metabolite profiling led to the identification of genes and key regulators involved in glucosinolate biosynthesis. Moreover, the integration of transcriptome and metabolome data of sulphur-deprived Arabidopsis plants revealed coordinate changes in the expression profiles of genes involved in glucosinolate and amino acid metabolism.This review provides an overview of our recent studies involving Arabidopsis mutant plants that exhibit impairment in the side-chain elongation process occurring during aliphatic glucosinolate biosynthesis by means of coexpression analysis and a novel metabolite profiling approach based on ultra-performance liquid chromatography coupled with tandem quadrupole mass spectrometry (UPLC-TQMS) (Sawada et al. 2009a). Thus, this review highlights the advantages of the omics-based approach in identifying genes involved in glucosinolate biosynthesis.

High-accuracy interferometer with a prism pair for measurement of the absolute refractive index of glass.

We propose a variable-path interferometric technique for the measurement of the absolute refractive index of optical glasses. We use two interferometers to decide the ratio between changes in the optical path in a prism-shaped sample glass and in air resulting from displacement of the sample. The method allows precise measurements to be made without prior knowledge of the properties of the sample. The combined standard uncertainty of the proposed method is 1.6x10(-6).

A chloroplastic UDP-glucose pyrophosphorylase from Arabidopsis is the committed enzyme for the first step of sulfolipid biosynthesis.

Plants synthesize a sulfur-containing lipid, sulfoquinovosyldiacylglycerol, which is one of three nonphosphorus glycerolipids that provide the bulk of the structural lipids in photosynthetic membranes. Here, the identification of a novel gene, UDP-glucose pyrophosphorylase3 (UGP3), required for sulfolipid biosynthesis is described. Transcriptome coexpression analysis demonstrated highly correlated expression of UGP3 with known genes for sulfolipid biosynthesis in Arabidopsis thaliana. Liquid chromatography-mass spectrometry analysis of leaf lipids in two Arabidopsis ugp3 mutants revealed that no sulfolipid was accumulated in these mutants, indicating the participation of UGP3 in sulfolipid biosynthesis. From the deduced amino acid sequence, UGP3 was presumed to be a UDP-glucose pyrophosphorylase (UGPase) involved in the generation of UDP-glucose, serving as the precursor of the polar head of sulfolipid. Recombinant UGP3 was able to catalyze the formation of UDP-glucose from glucose-1-phosphate and UTP. A transient assay using fluorescence fusion proteins and UGPase activity in isolated chloroplasts indicated chloroplastic localization of UGP3. The transcription level of UGP3 was increased by phosphate starvation. A comparative genomics study on UGP3 homologs across different plant species suggested the structural and functional conservation of the proteins and, thus, a committing role for UGP3 in sulfolipid synthesis.

A suppressive role of the prolyl isomerase Pin1 in cellular apoptosis mediated by the death-associated protein Daxx.

The death-associated protein Daxx is a multifunctional factor that regulates a variety of cellular processes, including transcription and apoptosis. Several previous reports have indicated that Daxx is induced upon oxidative stress and is then subjected to phosphorylation-based functional modification. However, the precise molecular events underlying these phosphorylation events remain largely unknown. We report in our current study that the peptidyl-prolyl isomerase Pin1 is highly overexpressed in malignant human gliomas and inhibits Daxx-mediated cellular apoptosis. The targeted inhibition of Pin1 by small interfering RNA in A172 glioblastoma cells significantly enhances the apoptotic response induced by hydrogen peroxide or stimulatory Fas antibodies. This is in turn accompanied by the increased induction of Daxx and the activation of the apoptosis signal-regulating kinase 1/c-Jun N-terminal kinase pathway. Furthermore, Pin1 binds to the phosphorylated Ser178-Pro motif in the Daxx protein, and Pin1 overexpression results in the rapid degradation of Daxx via the ubiquitin-proteasome pathway. Moreover, a Daxx-S178A mutant, which cannot interact with Pin1, demonstrates higher proapoptotic activity and is refractory to Pin1-mediated antiapoptotic effects. We further found that the expression levels of Pin1 inversely correlate with the degree of Daxx nuclear accumulation in human glioblastoma tissues. These results together indicate that Pin1-mediated prolyl isomerization plays an important role in the negative regulation of Daxx and thereby inhibits the oxidative stress-induced cellular apoptotic response, particularly in malignant tumor cells where Pin1 is often overexpressed.

Measurement of group refractive index wavelength dependence using a low-coherence tandem interferometer.

A technique for the measurement of the group refractive index wavelength dependence of optical materials using a low-coherence tandem interferometer and a spectrometer is proposed. Four channeled spectra resulting from interferences of light beams from different pairs of optical paths are used for the calculation of optical path differences. The group refractive index wavelength dependence is calculated from these optical path differences generated from the sample under measurement. No a priori information of the geometric thickness of a sample is required. The wavelength dependence of the group refractive index of the samples BK7 parallel plate of 5.200 and 10.025 mm from 675 to 850 nm is experimentally measured with an accuracy of the order of 10(-5) and a repeatability of the order of 10(-9).

Prolyl-isomerase Pin1 accumulates in lewy bodies of parkinson disease and facilitates formation of alpha-synuclein inclusions.

Parkinson disease (PD) is a relatively common neurodegenerative disorder that is characterized by the loss of dopaminergic neurons and by the formation of Lewy bodies (LBs), which are cytoplasmic inclusions containing aggregates of alpha-synuclein. Although certain post-translational modifications of alpha-synuclein and its related proteins are implicated in the genesis of LBs, the specific molecular mechanisms that both regulate these processes and initiate subsequent inclusion body formation are not yet well understood. We demonstrate in our current study, however, that the prolyl-isomerase Pin1 localizes to the LBs in PD brain tissue and thereby enhances the formation of alpha-synuclein immunoreactive inclusions. Immunohistochemical analysis of brain tissue from PD patients revealed that Pin1 localizes to 50-60% of the LBs that show an intense halo pattern resembling that of alpha-synuclein. By utilizing a cellular model of alpha-synuclein aggregation, we also demonstrate that, whereas Pin1 overexpression facilitates the formation of alpha-synuclein inclusions, dominant-negative Pin1 expression significantly suppresses this process. Consistent with these observations, Pin1 overexpression enhances the protein half-life and insolubility of alpha-synuclein. Finally, we show that Pin1 binds synphilin-1, an alpha-synuclein partner, via its Ser-211-Pro and Ser-215-Pro motifs, and enhances its interaction with alpha-synuclein, thus likely facilitating the formation of alpha-synuclein inclusions. These results indicate that Pin1-mediated prolyl-isomerization plays a pivotal role in a post-translational modification pathway for alpha-synuclein aggregation and in the resultant Lewy body formations in PD.

[Anesthetic management of tracheobronchial stent insertion in patients who underwent laser resection, balloon dilatation and tracheostomy in advance].

BACKGROUND: We report successful management of tracheobronchial stent insertion under general anesthesia. METHODS: In thirty-two cases, tracheobronchial stent insertion was performed under general anesthesia. The technique for airway management was chosen depending on the type of stent or the constriction level of the airway portion. We employed tracheostomy in order to avoid repeated intubations during the insertion of Dumon or Dynamic stent. In case of severe airway stenosis, laser resection or balloon dilatation was performed before stent insertion. RESULTS: We had 32 successful cases in 36 trials. Four trials failed due to insufficient expansion in one, mismatches of stent angle in one and pneumomediastinum in one. There was no exacerbation of respiratory condition in failed cases. There was no case who needed percutaneous cardiopulmonary support system. CONCLUSIONS: We managed tracheobronchial stent insertion under general anesthesia. Both the airway expansion by laser resection or balloon dilatation before stent insertion and also the insertion of Dumon or Dynamic stent through a tracheostomy were helpful strategies. These techniques facilitated more definitive airway maintenance and stable anesthetic management.

Low-coherence tandem interferometer for measurement of group refractive index without knowledge of the thickness of the test sample.

A new low-coherence interferometric technique is proposed for measuring the group refractive indices of dispersive samples with high accuracy. A tandem configuration of interferometers is used to compensate for the asymmetrical distortion of interferograms that results from the broad spectrum of the light source. The group refractive index can be measured without knowledge of the geometrical thickness of the sample under test. The proposed technique can successfully measure even a thick sample. Computer calculations have shown the effectiveness of the proposed technique, which was verified by preliminary experiments; the difference between the experimental result and the catalog data was 7 x 10(-4).

Broad-spectrum frequency comb generation and carrier-envelope offset frequency measurement by second-harmonic generation of a mode-locked fiber laser.

A frequency comb spanning more than one octave has been achieved by injecting the second-harmonic generation (780 nm) of a mode-locked fiber laser (1.56 microm) into a photonic crystal fiber. We propose and realize a novel interferometric scheme for observing the carrier-envelope offset frequency of the frequency comb. Frequency noise has been observed on the measured carrier-envelope offset frequency, which has been confirmed to be generated in the photonic crystal fiber by comparing the measured beat frequencies between cw lasers and frequency combs before and after the photonic crystal fiber. The mode-locked fiber laser is considered to be an important candidate for the light source used in realizing a compact optical frequency measurement system including applications in the telecommunication bands.

Heterodyne Fourier transform spectrometer for the near- infrared region.

A Fourier transform spectrometer with heterodyne modulation achieved by a moving diffraction grating has been developed for the near-infrared (NIR) region. The grating simultaneously acts as a beam splitter and a modulator, which realizes the optical frequency shift of incident light for increasing the sensitivity of measurements by the heterodyne detection technique. The differences in diffraction angle among broad spectra are compensated by a collimating mirror and plane mirrors. The proposed spectrometer is used for the measurements of spectra in the NIR region. The signal-to-noise ratio of measurements is improved sevenfold with a heterodyne modulation of 410 Hz. As examples, this spectrometer is applied for quantitative calibration and discrimination of organic solutions. The measurement of transmission spectra of a grape is also demonstrated.