东方Cheese发酵过程中异黄酮、苷元及组分的研究

东方Cheese经毛霉发酵能去除某些过敏源,这种无盐制品深受日本民众欢迎。文中通过正交试验优化了东方Cheese中异黄酮的提取条件。采用了三波长二标样法和HPLC测定异黄酮含量和组成。检测了东方Cheese发酵过程中异黄酮含量及组成的变化:异黄酮的总量降低了37.90%;其中糖苷由34.02μg/mL降至3.11μg/mL(即已有90.86%糖苷转化为苷元),苷元含量则相应的由0.97μg/mL升至18.62μg/mL。认为异黄酮总量降低以及组成变化的主要因素是糖苷到苷元分子质量的降低和样品含水量提高;发酵后期限制组成变化的主要因素是底物浓度的降低。     

Identification of the gene PaEMT1 for biosynthesis of mannosylerythritol lipids in the basidiomycetous yeast Pseudozyma antarctica

The yeast Pseudozyma antarctica produces a large amount of glycolipid biosurfactants known as mannosylerythritol lipids (MELs), which show not only excellent surface‐active properties but also versatile biochemical actions. To investigate the biosynthesis of MELs in the yeast, we recently reported expressed sequence tag (EST) analysis and estimated genes expressing under MEL production conditions. Among the genes, a contiguous sequence of 938 bp, PA_004, showed high sequence identity to the gene emt1, encoding an erythritol/mannose transferase of Ustilago maydis, which is essential for MEL biosynthesis. The predicted translation product of the extended PA_004 containing the two introns and a stop codon was aligned with Emt1 of U. maydis. The predicted amino acid sequence shared high identity (72%) with Emt1 of U. maydis, although the amino‐terminal was incomplete. To identify the gene as PaEMT1 encoding an erythritol/mannose transferase of P. antarctica, the gene‐disrupted strain was developed by the method for targeted gene disruption, using hygromycin B resistance as the selection marker. The obtained ΔPaEMT1 strain failed to produce MELs, while its growth was the same as that of the parental strain. The additional mannosylerythritol into culture allowed ΔPaEMT1 strain to form MELs regardless of the carbon source supplied, indicating a defect of the erythritol/mannose transferase activity. Furthermore, we found that MEL formation is associated with the morphology and low‐temperature tolerance of the yeast. Copyright © 2010 John Wiley & Sons, Ltd.     

Lingual Epithelial Stem Cells and Organoid Culture of Them

As tongue cancer is one of the major malignant cancers in the world, understanding the mechanism of maintenance of lingual epithelial tissue, which is known to be the origin of tongue cancer, is unquestionably important. However, the actual stem cells that are responsible for the long-term maintenance of the lingual epithelium have not been identified. Moreover, a simple and convenient culture method for lingual epithelial stem cells has not yet been established. Recently, we have shown that Bmi1-positive cells, residing at the second or third layer of the epithelial cell layer at the base of the interpapillary pit (IPP), were slow-cycling and could supply keratinized epithelial cells for over one year, indicating that Bmi1-positive cells are long-term lingual epithelial stem cells. In addition, we have developed a novel lingual epithelium organoid culture system using a three-dimensional matrix and growth factors. Here, we discuss current progress in the identification of lingual stem cells and future applications of the lingual culture system for studying the regulatory mechanisms of the lingual epithelium and for regenerative medicine.     

Space charge behavior in polymer film based on increment of capacitance under AC high field

Polymer materials have excellent dielectric and insulation properties; however, those properties in AC high field region have not been known well. Recently we established an evaluation method of high‐field AC dissipation current waveform of polymer materials 1 . AC dissipation current waveforms of polyethylene and polypropylene films show nonlinear distortion in AC high‐field region. This nonlinearity was thought to be related to the behavior of AC space charge formation in the sample near electrodes. The properties of space charge formed under AC high field at power frequency seem to differ from those formed under DC high field. The measurement of AC space charge distribution is not so easy due to the resolution limit of the space charge measurement. We studied the dielectric properties of biaxially oriented polypropylene (BOPP) film under AC high field up to 120 °C. It was found that tan δ, AC dissipation current (Ixr), and unbalanced component of capacitive current (ΔIxc) increased when the temperature became higher. In particular, ΔIxc increased above some threshold field and was considered to be due to the AC space charge formation. This AC space charge layer near electrode is thought to be formed due to carrier injection under AC high‐field application. Usually, the carrier mobility becomes smaller on lowering the temperature. Most of the carriers injected from the electrode are trapped near the electrode in the sample film. But in the high‐temperature region, the carrier mobility becomes larger and the carrier injection starts to increase from lower field. Many more carriers are injected from the electrode. It is thought that some of the injected carriers are trapped inside the sample film; the others go through the sample to the opposite side. © 2002 Wiley Periodicals, Inc. Electr Eng Jpn, 141(2): 8–16, 2002; Published online in Wiley InterScience ( www.interscience.wiley.com ). DOI 10.1002/eej.10018     

The protein encoded by cancer/testis gene D40/AF15q14 is localized in spermatocytes, acrosomes of spermatids and ejaculated spermatozoa

We have previously identified and cloned a human gene, D40, that is preferentially expressed in testis among normal organs, while it is widely expressed in various human tumor cell lines and primary tumors derived from different organs. In this report, we have examined the expression and localization of this protein in human testis with an antibody specific to D40 protein. In Western analyses, the anti-D40 antibody recognized a major band with a molecular mass of 300 kDa and a minor band of 250 kDa. These bands were not observed in the testis lysates from patients with Sertoli-cell-only syndrome and with Kleinfelter syndrome, who lack germ cells of the testis, indicating that D40 protein is expressed in the germ cells of normal testis. Immunohistochemical studies have revealed that D40 protein is highly expressed in spermatocytes and in the pre-acrosome of round spermatids. In the acrosome, D40 protein expression is observed not inside but outside the acrosome membrane. This is consistent with the finding that the amino-acid sequence at the amino terminal of the D40 protein lacks a hydrophobic signal peptide that is required for proteins to translocate to the membrane. Expression of D40 protein is observed in the acrosome of ejaculated spermatozoa as well, although the level is low compared with that in the pre-acrosome of spermatids. These results suggest that D40 protein plays important roles in spermatogenesis, especially in the formation and maintenance of the acrosome.     

Reaction of thiobarbituric acid with saturated aldehydes

The reaction of thiobarbituric acid (TBA) with saturated aldehydes, i.e., 1-butanal, 1-hexanal and 1-heptanal, produced a 455-nm yellow and a 532-nm red pigment. Formation of the pigments depended on the reaction conditions. The yellow pigment was unstable in the presence of excess amounts of the saturated aldehydes. The red pigment was formed only when the reaction was performed at a TBA/aldehyde ratio of 1∶1 in aqueous acetic acid. Formation of the yellow and red pigments required molecular oxygen. The colorless adducts, intermediates for the yellow and the red pigments, were isolated from the reaction mixtures. Aldol condensation and dehydration of 2 mol of the saturated aldehydes initially gave the α,β-unsaturated aldehydes, which in turn reacted with TBA to form the colorless adducts, pyranopyrimidine derivatives. The adducts were then converted into the yellow and red pigments under aerobic conditions.     

Complete erasing of ghost images on computed radiography plates and role of deeply trapped electrons

Computed radiography (CR) plates made of europium-doped Ba(Sr)FBr(I) were simultaneously exposed to filtered ultraviolet light and visible light in order to erase ghost images, i.e., latent image that is unerasable with visible light (LIunVL) and reappearing one, which are particularly observed in plates irradiated with a high dose and/or cumulatively over-irradiated. CR samples showing LIunVLs were prepared by irradiating three different types of CR plates (Agfa ADC MD10, Kodak Directview Mammo EHRM2, and Fuji ST-VI) with 50 kV X-ray beams in the dose range 8.1 mGy—8.0 Gy. After the sixth round of simultaneous 6 h exposures to filtered ultraviolet light and visible light, all the LIunVLs in the three types of CR plates were erased to the same level as in an unirradiated plate and no latent images reappeared after storage at 0 °C for 14 days. With conventional exposure to visible light, LIunVLs consistently remained in all types of CR plates irradiated with higher doses of X-rays and latent images reappeared in the Agfa M10 plates after storage at 0 °C. Electrons trapped in deep centers cause LIunVLs and they can be erased by simultaneous exposures to filtered ultraviolet light and visible light. To study electrons in deep centers, the absorption spectra were examined in all types of irradiated CR plates by using polychromatic ultraviolet light from a deep-ultraviolet lamp. It was found that deep centers showed a dominant peak in the absorption spectra at around 324 nm for the Agfa M10 and Kodak EHRM2 plates, and at around 320 nm for the Fuji ST-VI plate, in each case followed by a few small peaks. The peak heights were dose-dependent for all types of CR samples, suggesting that the number of electrons trapped in deep centers increases with the irradiation dose.     

Sedimentation study on the binding of fibrinogen or of antithrombin III with acidic polysaccharides including heparin

Summary The binding of fibrinogen with heparin and dextran sulfates, and of antithrombin III with heparin and dextran sulfate were investigated by sedimentation velocity method. From the measurement of fibrinogen-acidic polysaccharide systems, it was confirmed that heparin and low molecular weight dextran sulfate(DSC) forms soluble complexes with fibrinogen, though the amount bound of the latter was rather small compared with high molecular weight dextran sulfate(DSD). Antithrombin III was also found to be bound by heparin and DSC. The numbers of heparin molecules bound to one molecule of fibrinogen and antithrombin III were estimated to be 4.2 and 0.3, respectively.