As the second most abundant transition metal in humans, zinc plays essential roles in normal cellular biological functions, including metabolism, signalling, proliferation, gene expression and apoptosis. We use ZnSO₄ as a stressor in this study to investigate for the first time the effects of exogenous Zn²⁺ on both the cellular distribution of zinc and zinc-related proteins and the cell cycle of human lung adenocarcinoma (A549) cells. The cellular distribution of zinc and soluble proteins was determined in the whole cell as well as in the cytoplasmic and nuclear fractions. Exogenous zinc in the tested exposure range (0–100 µM) resulted in an altered cellular distribution of both zinc and the soluble proteins, together with total glutathione (GSx), the ratio of glutathione (GSH) to glutathione disulfide (GSSG) and non-protein sulphydryl (NPSH). Surprisingly, a turning point was observed in the re-distribution trend at a concentration of approximately 50 µM ZnSO₄. It is concluded that there exists a regulatory system in A549 cells that maintains the cellular zinc content stable in the presence of a certain range of extracellular zinc concentration. In addition, an MTT assay and flow cytometric analysis showed that the ZnSO₄ treatment led to a bi-phasic variation in viability and a slight fluctuation in the apoptosis of A549 cells. Our results will help to further elucidate zinc-related cell biology and biochemistry.

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An elemental analysis was performed on 30 Korean and Chinese kimchi samples in combination with strontium (Sr) isotope ratio measurements to develop a method for tracing the geographic origins of Korean and Chinese kimchi. We found a distinct difference between Korean and Chinese kimchi in such specific elements as K, Na, Ca, Ba, Sr, Zn, Li, and Sc, but no significant difference in the Sr isotope ratio. A comparative analysis was performed with washed kimchi (containing no submaterial) and unwashed kimchi to evaluate the effect of submaterials on the determination of geographic origin. A linear discriminant analysis provided a clear distinction between Korean and Chinese kimchi. The results indicate that combining multi-elemental and statistical analyses has the potential to determine the geographic origins of such processed food as kimchi.

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An enzymatic assay system for D-Asp was established using D-aspartate oxidase and oxaloacetate decarboxylase. In this system, D-Asp is converted to pyruvate, which is determined fluorometrically with 1,2-diamino-4,5-methylenedioxybenzene. This method makes possible D-Asp measurement at the micromolar level. The D-Asp contents of an edible brown alga, Hijika fusiforme, a lactic acid bacteria beverage, and pig testis were determined by the method.

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Scorpion venom is composed of a number of peptides, many of which show neurotoxicity. The Isometrus maculatus scorpion, belonging to the Buthidae family, is found in many tropical regions, including the southern islands of Japan, but there have been no reports on the isolation of toxins from its venom affecting sodium channels. We isolated in this study a novel toxin, Im-2, from the I. maculatus venom. Im-2 induced paralysis in crickets after injecting 20 µg/g of body weight. Im-2 also induced lethality in mice after an intracerebroventricular injection, indicating that Im-2 had non-selective toxicity between insects and mammals. Im-2 consists of 68 amino acids cross-linked by 4 disulfide bonds, and has sequence similarity to scorpion β-toxins that have been reported to affect the sodium channels of both insects and mammals. The toxic symptoms caused by Im-2 suggest that it acted on the nervous system and shared the action mechanism(s) with sequence-homologous β-toxins.

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Propolis collected on Jeju Island, Korea, contains characteristic components not present in propolis from other regions. Hence, the plant origin of the propolis from Jeju Island can be expected to be a novel plant. To identify the plant origin of this propolis, first we observed honeybee behavior, and found them collecting the resin from Angelica keiskei. Then comparative analyses of chemical and biological properties of the resin from the plant and propolis from hives of nearby apiaries were performed. Alcoholic extracts showed entirely identical HPLC profiles and closely similar antioxidant activities. These results indicate that A. keiskei is the plant origin of the propolis from Jeju Island, Korea.

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Larvae of the clearwing moth, Carmenta mimosa (Lepidoptera: Sesiidae), bore into the trunk of Mimosa pigra L., which is one of the most invasive weeds in Vietnam. GC-EAD and GC-MS analyses of a pheromone gland extract revealed that the female moths produced (3Z,13Z)-3,13-octadecadienyl acetate. A lure baited with the synthetic acetate alone successfully attracted C. mimosa males in a field test. While the addition of a small amount of the corresponding alcohol did not strongly diminish the number of captured males, a trace of the aldehyde derivative or the (3E,13Z)-isomer markedly inhibited the attractiveness of the acetate. The diurnal males were mainly attracted from 6:00 am to 12:00 am.

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Biotin is one of the most useful tags in (bio)analytical science due to its specific interaction with avidin, but is not easy to convert because of its low solubility in most solvents. Friedel-Crafts acylation of biotin acid chloride in triflic acid was examined, and the synthesized derivatives had stronger affinity to avidin than biotin in a binding assay using 2-(4'-hydroxyphenylazo)benzoic acid.

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Immunological function in chicks is greatly affected by estrogen treatment during embryogenesis, but the mechanism of the estrogen effect is not fully understood. To elucidate the effect of estrogen on immune function, we observed estrogen receptor expression in the thymus and bursa of chick embryos by immunohistochemistry. We compared the distribution of estrogen receptor-positive cells with that of keratin-positive epithelial cells. Intense expression of estrogen receptors was detected in thymic and bursal lymphocytes. In peripheral lymphocytes, ER mRNA was detected by RT-PCR analysis. The results of fluorescence-activated cell sorting analysis indicated that the estrogen receptor was expressed in the cytoplasm of the lymphocytes. Furthermore, intense expression of the estrogen receptor was also confirmed in thymic Hassall's corpuscles, bursal follicle-associated epithelial cells, and the bursal interfollicular epithelium. Our results indicate that estrogen affects the differentiation of thymic and bursal lymphocytes, suggesting that the underlying role for estrogen in immune function.

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LX519290, a synthetic derivative from a combinatorial chemistry library, has been screened for anti-atopic activity, but its biological activities have not yet been elucidated. To assess whether LX519290 has the potential to lessen 2,4-dinitrofluorobenzene-induced atopic dermatitis symptoms in mice, first we sensitized the skin in the dorsal neck of C57BL/6 mice and re-sensitized the ear skin to determine the ear thickness. Then, we tested to determine whether LX519290 affect atopic dermatitis symptoms in vivo. The results indicate that LX519290 significantly mitigated inflammation indications including ear thickness, total T-cell numbers, and eosinophils. Moreover, the treatment drastically inhibited the levels of mediators such as interleukin-17E and histamine by 52% and 37% of control, respectively. Taken together, the data suggest that LX519290 can alleviate atopic parameters in mice.

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Glutathione (GSH) is involved in abscisic acid (ABA)- and methyl jasmonate (MeJA)-induced stomatal closure in Arabidopsis thaliana. In this study, we examined the effects of GSH-decreasing chemicals, p-nitrobenzyl chloride (PNBC), iodomethane (IDM), and ethacrynic acid (EA), on ABA- and MeJA-induced stomatal closure in Arabidopsis. Treatments with PNBC, IDM, and EA decreased GSH contents in guard cells. Depletion of GSH by PNBC and IDM enhanced ABA- and MeJA-induced stomatal closure and inhibition of light-induced stomatal opening by ABA, whereas EA did not enhance either ABA- and MeJA-induced stomatal closure or inhibition of light-induced stomatal opening by ABA. Depletion of GSH did not significantly increase the production of the reactive oxygen species (ROS), cytosolic alkalization, or cytosolic Ca²⁺ oscillation induced by ABA and MeJA. These results indicate that depletion of GSH enhances ABA- and MeJA-induced stomatal closure without affecting ROS production, cytosolic alkalization, or cytosolic Ca²⁺ oscillation in guard cells of Arabidopsis.

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Obovatol has various biological activities, including anti-proliferative, neurotrophic, anti-fibrillogenic, anti-platelet, anti-fungal and anti-inflammatory activities. In this study, we investigated the effects of JJK694, a synthesized obovatol derivative, on rabbit platelet activation and its molecular mechanisms. JJK694 significantly inhibited washed rabbit platelet aggregation and serotonin secretion induced by collagen and arachidonic acid, but had little effect on thrombin- or U46619-induced aggregation. These results suggest that JJK694 selectively inhibits collagen- and arachidonic acid-mediated signaling. JJK694 also showed a concentration-dependent decrease in cytosolic Ca²⁺ mobilization, but it had no effect on arachidonic acid liberation. On the other hand, it significantly inhibited the formation of arachidonic acid metabolites, including thromboxane A₂ (TXA₂), prostaglandin D₂, and 12-hydroxy-5,8,10,14-eicosatetraenoic acid (12-HETE), by suppression of cyclooxygenase (COX)-1 and lipoxygenase (LOX) activities. These results indicate that JJK694 hasanti-platelet activities through inhibition of arachidonic acid metabolite production by suppression of COX-1 and LOX activities.

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RGD peptides are popular drug delivery tools in treating integrin αVβ3-expressing malignant tumors and tumor vasculature cells. We investigated the specific delivery and pharmacological potential of enantiomeric mitochondria-disruptive peptides (RLYLRIGRR-NH₂, RLRLRIGRR-NH₂, ALYLAIRRR-NH₂, and RLLLRIGRR-NH₂) after conjugation with an integrin αVβ3-homing peptide, cyclic pentameric RGD peptide. The cyclic RGD-conjugated mitochondria-disruptive peptides exhibited specific internalization, apoptosis induction, and cytotoxicity against integrin αVβ3-high-expressing human umbilical vein endothelial cells. Our findings indicate that these novel peptide complexes might prove good anti-angiogenesis reagents.

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The mechanism was investigated for macrophage recognition of cells oxidized by diamide, a thiol group-specific oxidizing reagent. Jurkat cells exposed to various concentrations of diamide were recognized by macrophages, the cells exposed to 25 µM diamide being best recognized. CD43, a major glycoprotein on the Jurkat cell surface, tended to form clusters upon diamide oxidization, and pretreating Jurkat cells with the anti-CD43 antibody inhibited macrophage binding. This indicates that macrophages appeared to recognize CD43. Sodium dodecyl sulfate polyacrylamide gel electrophoresis and a Western blot analysis of CD43 of the diamide-oxidized cells showed no increase in the amount of cross-linked CD43 compared with control cells, indicating that cross-linking of CD43 by a disulphide bond was not involved in the clustering. Both CD43 clustering and binding of the oxidized cells to macrophages was prevented by the caspase inhibitor, benzyloxycarbonyl-Val-Ala-Asp (OMe) fluoromethyl ketone (Z-VAD-fmk), suggesting that the oxidized and macrophage-bound cells were undergoing apoptosis. A closer examination revealed that the caspase-3 activity, chromatin condensation, and DNA fragmentation in Jurkat cells were all increased by oxidation. The macrophage receptor involved in the binding appeared to be the cell-surface protein, nucleolin; an anti-nucleolin antibody treatment inhibited the binding. These results suggest that thiol group-oxidized cells underwent early apoptosis and were recognized by nucleolin on macrophages as early apoptotic cells.

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Here, we demonstrated the involvement of the domains in Arabidopsis high-light responsive serine/arginine-rich (SR) and SR-like proteins, atSR30 and atSR45a, respectively, in subcellular and subnuclear distribution using a series of structural domain-deleted mutants. Judging from the localization of the transiently expressed domain-deleted mutants in onion epidermal cells, the C terminal low complexity domain rich in arginine-serine repeats (C-RS) domain of atSR30 appeared to be necessary for the nuclear localization. On the other hand, the N-terminal RS (N-RS) domain of atSR45a was necessary for the accurate nuclear localization, although the N- or C-RS domain alone was sufficient for the nuclear speckled organization. The phosphorylation of RS domains of atSR45a is irrelevant to the regulation of its localization. atSR45a and atSR30 were co-localized in the speckles, suggesting their collaborative roles in the regulation of alternative splicing events.

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A double mutant of Escherichia coli acetyl esterase (EcAE) with enhanced enzymatic activity was obtained by random mutagenesis using error-prone PCR and screening for enzymatic activity by observing halo formation on a tributyrin plate. The mutant contained Leu97Phe (L97F) and Leu209Phe (L209F) mutations. Single mutants L97F and L209F were also constructed and analyzed for kinetic parameters, as well as double mutant L97F/L209F. Kinetic analysis using p-nitrophenyl butyrate as substrate indicated that the k_cat values of L97F and L97F/L209F were larger than that of the wild-type enzyme, by 8.3-fold and 12-fold respectively, whereas no significant change was observed in the k_cat value of L209F. The K_m values of L209F and L97F/L209F were smaller than that of the wild-type enzyme, by 2.9-fold and 2.4-fold respectively, whereas no significant change was observed in the K_m value of L97F. These results indicate that a combination of an increase in k_cat values due to the L97F mutation and a decrease in K_m value due to the L209F mutation renders the k_cat/K_m value of the double mutant enzyme 29-fold higher than that of the wild-type enzyme.

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ΦC31 integrase, a site-specific recombinase, can effectively mediate the integration of foreign genes bearing an attB sequence into pseudo attP sites of genomes in human, mouse, and Drosophila cells. In this study, we measured ΦC31 integrase-mediated homologous recombination between attB and pseudo attP sites in sheep cells. The integration efficiency of the EGFP expression cassette with the attB sequence increased at least 2-fold in sheep fibroblasts. Three pseudo-attP sites were identified in the sheep genome, located in the intergenic regions on chromosomes 4, 13, and 7 respectively. Moreover, the transgene that was integrated at the three pseudo attP sites exhibited high levels of expression. Our study indicates that the ΦC31 integrase system provides an efficient integration tool for genetic engineering of the sheep genome.

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The replication and transcription of cucumber mosaic virus (CMV) are catalyzed by multi-protein complex RNA-dependent RNA polymerase (RdRp), which is composed of the viral-encoded 1a and 2a proteins with host factors. We have reported that the N-terminal region of the polymerase 2a protein, composed of 126 amino acids, is required for interaction with the helicase 1a protein, and that the phosphorylation of the region abrogated interaction with the 1a protein, suggesting a mechanism of resistance in host plants against viral infection. Here, we found that three protein 2a kinases, of 60, 55, and 38 kDa, co-purified with the tobacco membrane fraction in an in-gel kinase assay. By yeast two-hybrid library screening using the N-terminal 126 amino acids of 2a as a bait, we identified CBL-interacting protein kinase 12 (NtCIPK12) corresponding to 55 kDa protein 2a kinase. The bacterially expressed protein kinase showed protein 2a kinase (t2aK) activity in vitro. We found that NtCIPK12 stabilized upon CMV infection at the post-translational level, and accumulated more heavily to the membrane than in the cytosol.

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We found that stem-cell leukemia (SCL), also known as T cell acute-lymphocytic leukemia (Tal-1) gene expression, was upregulated in the maturing rat testis. Strong expression of Tal-1 was detected in the normal maturing rat testis by Northern blotting. Western blotting revealed the protein size to be about 34 kDa. Protein expression was wide-spread in spermatocytes, spermtids and spermatogonia in accordance with the seminiferous epithelium cycle, as determined by an analysis of immunohistochemistry. Gene expression of Tal-1 regulatory gene, NKX3.1, was negatively correlated with Tal-1 expression. Human Tal-1 expression in the maturing testis as well as in bone marrow was observed, which suggests that the gene product is a novel cancer-testis antigen candidate. Taken together, TAL-1 may be involved in cell division, morphological changes, and the development of spermatogenic cells in the normal rat testis.

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Screening of rice full-length cDNA overexpressing (FOX) lines allowed the identification of a TIFY gene, TIFY11b, as a growth-promoting gene whose overexpression increased plant height and seed size. The grains of TIFY11b-overexpressing plants exceeded those of non-transformants in length, width and thickness, resulting in 9–21% increases in grain weight. The increase was achieved by overexpressing the gene in the whole plant body, but not by seed-restricted expression, indicating that seed enlargement is attributable to overexpression in vegetative organs such as the leaf. The whole-body overexpressing plants developed longer leaves along with higher levels of starch and sucrose in the leaf sheath and culm at the heading stage than the non-transformants. Although overexpression of TIFY11b did not alter the photosynthetic rate per leaf area before and after heading, it caused an accumulation of higher levels of the carbohydrate assimilate, probably due to increased photosynthesis per plant, suggesting that the increase in grain size and weight is attained by enhanced accumulation and translocation of the carbohydrate in the culms and leaf sheaths of the transgenic plants. Thus, TIFY11b is a novel grain-size increasing gene.

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The bean bug (Riptortus clavatus) is a serious insect pest of soybean. Corn (maize) cystatin strongly inhibited the activity of its digestive cysteine proteinase. Heterologous expression of corn cystatin in soybean seeds inhibited the insect's proteases, but not its growth.

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Efficiency of gene targeting was increased more than 10-fold when an excess molar ratio of non-homologous fragments was added in the transformation of Aspergillus nidulans. In the targeted transformants, integration of such fragments into the host chromosome was a rare event. Hence, our approach proved practical in terms of producing successful targeting events without disturbing the host chromosomes.

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NtP4H1.1 is a Golgi-localizing type II integral membrane protein. Mutations in the cytoplasmic tail direct the protein to the endoplasmic reticulum (ER). We expressed a GFP fusion protein containing the mutant tail and the transmembrane region in tobacco BY-2 cells, and found that the protein localized in the Golgi. Therefore NtP4H1.1 contains multiple targeting information in different regions.

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Mammalian cells express two isoforms of eIF5A, eIF5A1 and eIF5A2, but little is known about the function of eIF5A2. Here we report that eIF5A2 is reversibly acetylated at lysine-47. HDAC6 and SIRT2 were identified as the enzymes responsible for deacetylating eIF5A2. Analysis using acetylation-deficient mutants indicated that acetylation regulates the subcellular localization of eIF5A2.

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The insulin/insulin-like growth factor-1 signaling pathway of Caenorhabditis elegans regulates larval diapause and adult lifespan through the sole insulin receptor-like protein, DAF-2. In the present study, the physiological function and expression pattern of INS-17, one of the C. elegans insulin-like peptides, were examined by disruption and overexpression of the gene, and by the use of a reporter gene. INS-17 might function as a DAF-2 antagonist in the regulation of larval diapause, but not of the adult lifespan. The reporter protein was intensively expressed during larval diapause. It showed a drastic decrease in amount after larval diapause, which matches well the physiological function of INS-17.

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Recent studies have indicated that Listeria monocytogenes formed biofilms on the surface of food processing equipment, and may survive sanitization treatments. The purpose of this study was to compare the susceptibility of L. monocytogenes grown in either a biofilm or planktonic culture when exposed to hydrogen peroxide (H₂O₂). Twelve strains of biofilm-forming L. monocytogenes and their planktonic counterparts were treated with various concentrations of H₂O₂ (1, 6, and 10%), and the cell survival was then determined at 10-min exposure intervals. When grown as a biofilm, L. monocytogenes was significantly more resistant to H₂O₂ than under planktonic culture conditions. Planktonic L. monocytogenes strains exhibited significantly different susceptibility to 1% H₂O₂. Equally interestingly, biofilms of the 12 L. monocytogenes strains also inhibited different survival rates after being treated with 6 and 10% H₂O₂. However, most of the biofilms recovered to a population of 2–9 log CFU/glass fiber filter (GFF) after a 24-h re-growth period. These results indicate that there was no significant correlation between the H₂O₂ resistance of biofilm- and planktonic-cultured cells, and suggest that different mechanisms for the resistance to sanitation or disinfection underly the persistence of certain strains in food-processing environments.

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We investigated the adverse effects of Japanese butterbur leaves (Petasites japonicus, Compositae) in male F344/DuCrj rats. The rats were fed a control diet or a treatment diet containing 5% butterbur leaf powder for 4 weeks. No differences were observed in body weight gain, food intake or feed efficiency between treatments, but relative liver weight in the butterbur group was significantly higher than that of the control group. In addition, thiobarbituric acid reactive substances (TBARs) and glutathione levels in the serum and liver of the butterbur group were higher than those of the control group. Hepatic glutathione reductase and glutathione S-transferase activities and mRNA expression in the butterbur leaf group were higher than in the control group. Furthermore, hepatic cytochrome 2E1 mRNA expression was higher than in the control group. In vitro, an acetone extract of the butterbur leaf powder showed the strongest increase in TBARs level in a hepatic homogenate through 4 d. Our findings suggest that feeding 5% butterbur leaf powder to rats can cause adverse effects by increasing oxidative stress.

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We have previously reported that Aloe vera gel had hypoglycemic activity and anti-obesity effects, although the effect on alcoholic fatty liver was unclear. We examined in this present study the effect of an Aloe vera gel extract (AVGE) on hepatic lipid metabolism by using an ethanol-induced transient fatty liver mouse model. Ethanol (3 g/kg of mouse weight) was orally administered to induce an accumulation of triglyceride (TG) and increase the mRNA expression of such lipogenic genes as sterol regulatory element-binding protein-1 (SREBP-1) and fatty acid synthase (FASN) in the liver. Although ethanol ingestion caused a 5.4-fold increase in liver TG, pre-treating with AVGE (1 mg/kg/d) for 1 week significantly suppressed this elevation of the ethanol-induced liver TG level. The expression of lipogenic genes was also lower in the AVGE pre-treatment group than in the control group. This inhibitory effect on the ethanol-induced accumulation of TG was attributed to a reduction in the expression of lipogenic genes that were increased by ethanol.

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In lactating mammary glands, milk fat is secreted as fat globules surrounded by a cell plasma membrane containing characteristic membrane-associated proteins. Among these, butyrophilin has been shown to be specific and intrinsic to the fat globule membrane, whereas milk-fat globule EGF-factor VIII (MFG-E8) is uncertain. We characterized in the present study MFG-E8 in milk fat globules and in the culture medium of HC11 mammary epithelial cells. MFG-E8 was immunologically detected in the mammary tissues of both pregnant and lactating mice. Double-immunofluorescence staining for MFG-E8 and butyrophilin showed diversity in the MFG-E8-staining intensity among different fat globules in milk. HC11 cells secreted monomeric MFG-E8 with phosphatidylserine-binding activity, despite no fat globules being detected in the cells. This secretion was upregulated by not only prolactin but also by insulin or EGF. These results suggest that milk MFG-E8 was not equally present among fat globules and not necessarily intrinsic to the fat globules.

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We investigated the extracellular production of folate, vitamin B₁₂, and thiamine in cultures of lactic acid bacteria (LAB) isolated from nukazuke, a traditional Japanese pickle, and the relationships between the vitamin production and such properties of LAB as tolerance to salts, ethanol, etc. Among the 180 isolates of LAB, two strains of Lactobacillus (Lb.) sakei and a strain of Lb. plantarum extracellularly produced high levels of folate (about 100 µg/L). A strain of Lb. coryniformis and one of Lb. plantarum produced about 2 µg/L of vitamin B₁₂, although the level was not high. No isolates produced a high level of thiamine. The type cultures of LBA (53 strains) did not show any higher production of these vitamins. Some isolates showed tolerance to high concentrations of salts and alcohol, and low initial pH. No significant relationships between folate or vitamin B₁₂ productions and these properties of LAB were apparent.

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Royal jelly (RJ), a honeybee-derived product, has been found to possess developmental and physiological bioactivity in the fruit fly, Drosophila melanogaster, but little is known about the in vivo bioactivity of freeze-dried RJ (FDRJ) powder, which is another form of RJ processed for human use. To address this, we used Drosophila as a model animal to examine the effects of FDRJ in multicellular organisms. When flies were reared on food supplemented with FDRJ, the developmental time from larva to adult was shortened, the adult male lifespan was prolonged, and female fecundity was increased without any significant morphological alterations. Moreover, the expression of dilp5, an insulin-like peptide, its receptor InR, and the nutrient sensing molecule TOR, the target of rapamycin, was significantly increased in FDRJ-fed female flies as compared with ones reared on standard and on protein-enriched food. These findings suggest that like RJ, FDRJ maintains its bioactivity even after processing from RJ, what is expected to have bioactivity for multicellular organisms, including humans.

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A wx/ae double-mutant rice is generated by crossing waxy mutant and amylose-extender mutant in rice. Wx/ae brown rice contains highly beneficial nutrients for lipid and glucose metabolism, including resistant starch, dietary fiber, and γ-oryzanol, when compared to Koshihikari brown rice, the non-waxy japonica cultivar. To examine the effects of wx/ae brown rice on glucose and lipid metabolism, type 2 diabetic NSY/Hos mice were fed a high-fat diet containing 25% of wx/ae brown rice or Koshihikari brown rice for 10 weeks. The plasma total cholesterol, non-high-density lipoprotein cholesterol, triglyceride, and non-esterified fatty acid levels of the wx/ae group were significantly lower than those of the Koshihikari group. Moreover, the fasting blood glucose level and pathological score of glycosuria of the wx/ae group were also significantly lower than those of the Koshihikari group. These results indicate that wx/ae brown rice has the potential to prevent the rise in plasma lipid and glucose levels.

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β-Cryptoxanthin (β-CRX) is a carotenoid abundantly present in the Satsuma mandarin (Citrus unshiu Marc.), one of the most popular fruits in Japan, and it is reported to have several health benefits. Although it is thought to have higher bioavailability than other carotenoids, the usual daily intake is small, and a good method to improve its bioavailability is needed. Hence we studied the effect of emulsification on the absorption characteristics of β-CRX in the intestine. Human trials showed that its serum transfer efficiency was statistically higher in the emulsified formulation than in fresh Satsuma mandarin juice. Caco-2 permeability studies indicated that emulsifiers preferentially accelerate the absorption of the non-esterified form of β-CRX, suggesting that emulsification is more effective for free β-CRX. This information might be useful to improve the efficiency of β-CRX serum transfer, as well as to increase the health benefits of β-CRX.

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A novel method in which the water sorption curve is observed under linearly temperature-raising conditions was proposed to estimate the gelatinization temperature of starch-containing foods, it was applied in an estimation of the gelatinization temperatures of dried noodles. The gelatinization temperatures of two kinds of spaghetti, dried at high and low temperature, were 52.3 and 53.1 °C, and those of udon, kishimen, juwari-soba, hachiwari-soba, so-called common soba, Malony^®, and kuzukiri were 57.0, 57.8, 61.1, 59.6, 57.4, 48.4, and 49.1 °C. The gelatinization temperatures estimated by the method were between the onset and peak temperatures obtained by differential scanning calorimetric measurement.

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To increase isobutanol production in Saccharomyces cerevisiae, the valine biosynthetic pathway was activated by overexpression of the relevant enzymes in the mitochondria and the cytosol. Native mitochondrial enzymes were overepxressed in the cytosol by deleting the mitochondrial transit peptides. The metabolically engineered S. cerevisiae possessing the cytosolic pathway showed increased isobutanol production (63 ± 4 mg/L).

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