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  • Reconstitution of biosynthetic machinery of fungal natural products in heterologous hosts.
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : 2019-11-19
    Hideaki Oikawa

    Ascomycota and basidiomycota fungi are prolific sources of biologically active natural products. Recent genomic data and bioinformatic analysis indicate that fungi possess a large number of biosynthetic gene clusters for bioactive natural products but more than 90% are silent. Heterologous expression in the filamentous fungi as hosts is one of the powerful tools to expression of the silent gene clusters. This review introduces recent studies on the total biosynthesis of representative family members via common platform intermediates, genome mining of novel di- and sesterterpenoids including detailed cyclization pathway, and development of expression host for basidiomycota genes with efficient genome editing method. In addition, this review will discuss the several strategies, for the generation of structural diversity, which are found through these studies.

    更新日期:2020-01-10
  • His-Cys and Trp-Cys cross-links generated by post-translational chemical modification.
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : null
    Nobutaka Fujieda

    Galactose oxidase and amine oxidase contain a cofactor which is generated by post-translational chemical modification to the corresponding amino acid side chains near the copper active center. Such cofactors provide proteins unusual catalytic ability that canonical amino acids cannot exert as well as their structural stability, and thereby are called as protein-derived cofactors. These cofactors and modifications are mostly derived from aromatic amino acid residues, especially Tyr, Trp, and His. Current information about unusual cofactors derived from two of those, heteroaromatic residues (Trp and His) is summarized, especially chemical properties and maturation process of the cross-links between cysteine and heteroaromatic amino acids (His-Cys and Trp-Cys cross-links).Abbreviations: FMN: flavin mononucleotide; FAD: flavin adenine nucleotide; RNA: ribonucleic acid; PDC: protein-derived cofactor; GFP: green fluorescent protein; MIO: 3,5-dihydro-5-methylidene-4-imidazol-4-one; LTQ: lysyl tyrosylquinone; CTQ: cysteine tryptophylquinone; TTQ: tryptophan tryptophylquinone; E.coli: Escherichia coli; WT: wild type.

    更新日期:2020-01-10
  • Euphausia pacifica as a source of 8(R)-hydroxy-eicosapentaenoic acid (8R-HEPE), 8(R)-hydroxy-eicosatetraenoic acid (8R-HETE) and 10(R)-hydroxy-docosahexaenoic acid (10R-HDHA).
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : 2019-11-19
    Hidetoshi Yamada,Kanako Kumagai,Aiko Uemura,Sayaka Yuki

    Although not fully investigated, 8-HEPE, 8-HETE, and 10-HDHA have potentially beneficial effects for human health. Euphausia pacifica (North Pacific krill) is unique in containing several ppm level of 8R-HEPE, and sub-ppm levels of 8R-HETE and 10R-HDHA. Obtaining sufficient quantities of these compounds is a major bottleneck for conducting in vivo experiments to evaluate their biological activities. In this study, we examined an efficient way of obtaining 8R-HEPE, 8R-HETE, and 10R-HDHA by enzymatic production in E. pacifica. We devised a novel method to purify 199.4 mg of 8R-HEPE, 2.1 mg of 8R-HETE and 5.6 mg of 10R-HDHA from 1 kg of E. pacifica. We identified the stereochemistry of the hydroxy group at C-8 of HEPE and HETE and C-10 of HDHA as the R configuration by chiral column chromatography analysis using LC/QTOFMS.Abbreviations: 8-HEPE: 8-hydroxy-eicosapentaenoic acid; 8-HETE: 8-hydroxy-eicosatetraenoic acid; 10-HDHA: 10-hydroxy-docosahexaenoic acid; EPA: eicosapentaenoic acid; TLC-FID, thin layer chromatograph-Flame Ionization Detector; LC/QTOFMS: liquid chromatography/hybrid quadrupole time of flight mass spectrometry.

    更新日期:2020-01-10
  • Production of lycopene by metabolically engineered Pichia pastoris.
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : null
    Xinying Zhang,Denggang Wang,Yehong Duan,Xueyun Zheng,Ying Lin,Shuli Liang

    Lycopene is a highly valued carotenoid with wide applications in various industries. The market demand for lycopene promotes research in metabolic engineering of heterologous hosts for lycopene. In this study, Pichia pastoris strain GS115 was genetically engineered to produce lycopene by integrating the heterologous lycopene biosynthesis genes from Corynebacterium glutamicum ATCC13032. The resulting strain, L1, produced 0.115 mg/g cell dry weight (DCW) lycopene. Through optimization by promoter selection, improving the precursor supply and expanding the Geranylgeranyl diphosphate (GGPP) pool, ultimately, the lycopene yield of the final optimal strain was 6.146 mg/g DCW with shake flask fermentation and 9.319 mg/g DCW (0.714 g/L) in a 3 L fermenter. The lycopene yield in this study is the highest yield of lycopene in P. pastoris reported to date, which demonstrated the potential of P. pastoris in lycopene synthesis and as a candidate host organism for the synthesis of other high value-added terpenoids.

    更新日期:2020-01-10
  • Tryptophanyl-tRNA synthetase (WARS) expression in uveal melanoma - possible contributor during uveal melanoma progression.
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : 2019-11-07
    Pan-Pan Yang,Xiao-Hui Yu,Jiao Zhou

    This study aimed to explore the influence of Tryptophanyl-tRNA synthetase (WARS) expression on the proliferation and migration of uveal melanoma (UM) cells, and the potential mechanisms. Bioinformatics analysis based on Gene Expression Omnibus (GEO) database showed that WARS expression in metastatic cancer was significantly higher than that in no-metastatic group. Kaplan-Meier analysis based on The Cancer Genome Atlas (TCGA) database showed that high WARS expression was associated with lower survival. Biological function experiments showed that overexpression of WARS in OCM-1A cells can promote cell proliferation, migration, and invasion, whereas knockdown of WARS in C918 cells showed the opposite effect. Finally, we observed that the up-regulation of WARS induced the activation of phosphatidylinositol 3-kinase/AKT (PI3K/AKT) signaling, whilst depletion of WARS resulted in opponent outcomes. Taken together, our results illustrated that WARS was overexpressed in UM cells and contributed to the viability and motility of UM cells via modulating PI3K/AKT signaling pathway.

    更新日期:2020-01-10
  • Super-activator variants of the cyanobacterial transcriptional regulator ChlR essential for tetrapyrrole biosynthesis under low oxygen conditions.
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : null
    Yuto Hiraide,Haruki Yamamoto,Yasushi Kawajiri,Hisanori Yamakawa,Kei Wada,Yuichi Fujita

    ChlR is a MarR-type transcriptional regulator that activates the transcription of the chlAII-ho2-hemN operon in response to low oxygen conditions in the cyanobacterium Synechocystis sp. PCC 6803. Upon exposure to low oxygen conditions, ChlR activates transcription of the operon that encodes enzymes critical to tetrapyrrole biosynthesis under low oxygen conditions. We previously identified a super-activator variant, D35H, of ChlR that constitutively activates transcription of the operon. To gain insight into the low-oxygen induced activation of ChlR, we obtained eight additional super-activator variants of ChlR including D35H from pseudorevertants of a chlAI-disrupted mutant. Most substitutions were located in the N-terminal region of ChlR. Mapping of the substituted amino acid residues provided valuable structural insights that uncovered the activation mechanism of ChlR.

    更新日期:2020-01-10
  • Overexpressing of POU2F2 accelerates fracture healing via regulating HMGA1/Wnt/β-catenin signaling pathway.
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : null
    Wei Zhang,Hanke Zhao,Kun Chen,Ye Huang

    To elucidate the role of POU2F2 (POU class 2 homeobox 2) in fracture healing, 30 rats with femoral fracture were randomly grouped into three groups: FF group, LV-POU2F2 group and LV-scramble group. Rats were injected with PBS, lentivirus expressing POU2F2 or scramble lentivirus once a week for 4 weeks. Results showed that overexpressing of POU2F2 promoted fracture healing and callus growth. Besides, overexpressing of POU2F2 promoted protein and mRNA expression of Col10a1, Runx2, Osterix, and Osteocalcin. High Mobility Group AT-hook 1 (HMGA1) is a non-histone protein participating in chromatin remodeling of cells. Western blotting manifested HMGA1/Wnt/β-catenin pathway was activated in POU2F2 group. Moreover, in-vitro study of hMSCs cells supported the above data. In conclusion, POU2F2 promotes fracture healing via activating the HMGA1/Wnt/β-catenin signaling pathway.

    更新日期:2020-01-10
  • Liquid chromatography-electrospray ionization-tandem mass spectrometric assay for d-aspartate N-methyltransferase activity in ark shells.
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : 2019-11-07
    Kimihiko Shibata,Saho Hagiya,Rena Okawara,Katsumasa Abe,Shouji Takahashi,Yoshio Kera

    A liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) method for the separation and quantification of the enantiomers of N-methylaspartate and N-methylglutamate, after derivatization with Nα-(5-fluoro-2,4-dinitrophenyl)-L-leucinamide was established. The time required for the LC-ESI-MS/MS analysis was within 20 min and the detection limit was approximately 10 fmol per injection, demonstrating that this method can be used for the rapid determination of D-aspartate N-methyltransferase activity in the ark shell clam Scapharca broughtonii.Abbreviations: NMDA: N-methyl-D-aspartate; NMLA: N-methyl-L-aspartate; NMDG: N-methyl-D-glutamate; NMLG: N-methyl-L-glutamate; NMA: N-methylaspartate; NMG: N-methylglutamate; HPLC: high-performance liquid chromatography; SAM: S-adenosyl-L-methionine; OPA: o-phthalaldehyde; LC-ESI-MS/MS: liquid chromatography-electrospray ionization-tandem mass spectrometry; FDLA: Nα-(5-fluoro-2,4-dinitrophenyl)-L-leucinamide; FDAA: Nα-(5-fluoro-2,4-dinitrophenyl)-L-alaninamide; ESI: electrospray ionization; LC-ESI-MS: liquid chromatography-electrospray ionization-mass spectrometry; MS/MS: tandem mass spectrometry.

    更新日期:2020-01-10
  • Oxidation of isomaltose, gentiobiose, and melibiose by membrane-bound quinoprotein glucose dehydrogenase from acetic acid bacteria.
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : null
    Takaaki Kiryu,Taro Kiso,Hirofumi Sato,Hiromi Murakami

    Membrane-bound quinoprotein glucose dehydrogenase from acetic acid bacteria produces lactobionic acid by the oxidation of lactose. Its enzymatic activity on lactose and maltose is much lower than that on D-glucose. For that reason, the activity of the enzyme on disaccharides has been considered low. In this study, we show that the isomaltose-oxidizing activity of acetic acid bacteria is much higher than their lactose-oxidizing activity. In addition to isomaltose, the enzyme oxidized gentiobiose and melibiose to the same extent. According to the characteristics of the isomaltose-oxidizing activity and investigations using dehydrogenase-deficient mutant bacteria, we identified the responsible enzyme as membrane-bound quinoprotein glucose dehydrogenase.Abbreviations: AAB: acetic acid bacteria; m-GDH: membrane-bound quinoprotein glucose dehydrogenase; DCIP: 2,6-dichlorophenolindophenol; DP: degree of polymerization; HPAEC-PAD: high-performance anion-exchange chromatography with pulsed amperometric detection; NMR: nuclear magnetic resonance; TLC: thin layer chromatography; COSY: correlation spectroscopy.

    更新日期:2020-01-10
  • Inhibition of melanin production and promotion of collagen production by the extract of Kuji amber.
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : null
    Satoshi Suzuki,Junpei Abe,Yui Kudo,Masayuki Shirai,Ken-Ichi Kimura

    Kuji amber is fossilized tree resin of the Late Cretaceous in Japan. In this study, new biological activities of ethanol extract of Kuji amber (EtOH ext.) and supercritical carbon dioxide fluid extract of Kuji amber (scCO2 ext.) were examined. Both EtOH ext. and scCO2 ext. inhibited melanin production in B16 mouse melanoma cells and promoted collagen production in human skin fibroblast SF-TY cells. The scCO2 ext. had more potent activity than that of EtOH ext. and may depend on the efficiency of the extraction. The main new biologically active compound in Kuji amber, kujigamberol had no activities against melanin production, however, it promoted collagen production at low concentrations. A biologically active compound having a different structure, spirolactone norditerpenoid, showed both the inhibition activity against melanin production and the promotion activity of collagen production in a dose dependent manner. EtOH ext. and scCO2 ext., which include both kujigamberol and spirolactone norditerpenoid, have not only anti-allergy activity, but also inhibit melanin production and promote collagen production.

    更新日期:2020-01-10
  • Tocotrienol-rich fraction from annatto ameliorates expression of lysyl oxidase in human osteoblastic MG-63 cells.
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : 2019-11-20
    Kakeru Kohno,Wakana Yamada,Aya Ishitsuka,Miki Sekine,Nantiga Virgona,Masako Ota,Tomohiro Yano

    Lysyl oxidase (LOX) is required for the formation of bone collagen cross-links. Inactivation of the LOX gene in osteoblasts by DNA methylation and JAK signaling has been reported to cause loss of cross-links and an increased risk of fractures. Tocotrienols (T3s) have proven benefits on bone strength, but their potential effects on LOX remain largely unknown. Thus, the present study investigates the in vitro effects of T3s on LOX expression in human osteoblastic MG-63 cells. Results indicated that Tocotrienol-Rich Fraction (TRF), the δ-T3 rich oil extracted from Annatto was the most effective and significantly increased LOX expression. TRF treatment decreased de-novo methyltransferases (DNMTs), DNMT3A and DNMT3B levels. In addition, TRF significantly inhibited JAK2 activation and decreased expression of Fli1, a transcription factor of DNMTs. We conclude that TRF induced an increase in LOX expression via inhibition of de-novo methylation and reduction of Fli1 expression by the inactivation of JAK2.Abbreviations: CpG: cytosine-guanine dinucleotide; DNMT: DNA methyltransferase; Fli1: friend leukemia virus integration 1; JAK: janus kinase; LOX: lysyl oxidase; PCR: polymerase chain reaction; STAT: signal transducers and activators of transcription; T3s: tocotrienols; TPs: tocopherols; TRF: Tocotrienol-Rich Fraction.

    更新日期:2020-01-10
  • Olive polyphenol reduces the collagen-elicited release of phosphorylated HSP27 from human platelets.
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : null
    Daisuke Mizutani,Takashi Onuma,Kumiko Tanabe,Akiko Kojima,Kodai Uematsu,Daiki Nakashima,Tomoaki Doi,Yukiko Enomoto,Rie Matsushima-Nishiwaki,Haruhiko Tokuda,Shinji Ogura,Hiroki Iida,Osamu Kozawa,Toru Iwama

    Hydroxytyrosol (HT) and oleuropein (OLE) are natural polyphenols found in extra virgin olive oil. Accumulating evidence indicates that ingestion of olive oil contributes to reduce the risk of cardiovascular diseases and stroke. It has been reported that HT and OLE inhibit human platelet aggregation. We have shown that collagen induces the phosphorylation of heat shock protein 27 (HSP27) in human platelets, resulting in the release of HSP27, an extracellular pro-inflammatory agent. In this study, we investigated the effects of HT and OLE on the collagen-stimulated human platelet activation. The PDGF-AB secretion and the soluble CD40 ligand (sCD40L) release by collagen were reduced by HT or OLE. HT and OLE significantly suppressed the phosphorylation of HSP27 and the release of phosphorylated-HSP27. These findings suggest that olive polyphenol reduces the collagen-stimulated phosphorylation of HSP27 in human platelets and the release. Our results may provide a novel anti- inflammatory effect of olive polyphenol.

    更新日期:2020-01-10
  • Effects of genistein on lipopolysaccharide-induced injury of mouse alveolar epithelial cells and its mechanism.
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : null
    Qi Zhu,Wei Zhang,Deguang Mu,Hongbin Zhou,Shengchang Wu,Hai Zou

    Alveolar and bronchial epithelial cells have critical functions in acute respiratory distress syndrome progress. Genistein could protect the lungs from acute lung injury, however, whether genistein protects the alveolar epithelial cells from LPS-induced injury was less studied. Spectrophotometric method 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) and enzyme-linked immunosorbent assay (ELISA) were performed to detect cell viability and levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and IL-6. Flow cytometry and western blot assay were performed to detect cells apoptosis and protein levels. In LPS-induced model of mouse lung epithelial (MLE)-12 cells, PBEF (proinflammatory cytokine) expression, and cell apoptosis were increased and cell viability was decreased, whereas NF-κB was activated and expression levels of TNF-α, IL-1β, and IL-6 were increased. However, genistein partly reversed the effect of LPS, and it plays a protective role in lung injury by reducing expression of PBEF, inhibiting the activation of NF-κB and alleviating inflammatory response of cells.

    更新日期:2020-01-10
  • The plant-type phosphoenolpyruvate carboxylase Gmppc2 is developmentally induced in immature soy seeds at the late maturation stage: a potential protein biomarker for seed chemical composition.
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : null
    Naoki Yamamoto,Toshio Sugimoto,Tomoyuki Takano,Ai Sasou,Shigeto Morita,Kentaro Yano,Takehiro Masumura

    Phosphoenolpyruvate carboxylase (PEPC) is a carbon-fixing enzyme with critical roles in seed development. Previously we observed a positive correlation between PEPC activity and protein content in mature seeds among soybean cultivars and varietal differences of PEPC activity in immature seeds, which is concordant with seed protein accumulation. Here, we report a PEPC isoform (Gmppc2) which is preferentially expressed in immature soybean seeds at the late maturation stage. Gmppc2 was co-expressed with enzyme genes involved in starch degradation: α-amylase, hexokinase, and α-glucan phosphorylase. Gmppc2 was developmentally induced in the external seed coats, internal seed coats, hypocotyls, and cotyledons at the late maturation stage. The expression of Gmppc2 protein was negatively regulated by the application of a nitrogen fertilizer, which suppressed nodule formation. These results imply that Gmppc2 is involved in the metabolism of nitrogen originated from nodules into seeds, and Gmppc2 might be applicable as a biomarker of seed protein content.Abbreviations: PEP: phosphoenolpyruvate; PEPC: phosphoenolpyruvate carboxylase; RNA-Seq: RNA sequencing; PCA: principal component analysis; SE: standard error.

    更新日期:2020-01-10
  • Purification, cDNA cloning and characterization of Kunitz-type protease inhibitors from Apios americana tubers.
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : null
    Jia Liu,Masami Yonekura,Yoshiaki Kouzuma

    Two kinds of Kunitz-type protease inhibitors, AKPI1 and AKPI2, were purified from Apios americana tubers by four steps of column chromatographies and their cDNA cloning was performed. AKPI1 cDNA consist of 809 nucleotides, and the matured protein had 190 amino acids with 20,594 Da. AKPI2 cDNA consist of 794 nucleotides, and the matured protein had 177 amino acids with 19,336 Da. P1 site of AKPI2 was Leu88, suggested the target enzyme was chymotrypsin. On the other hand, Gly85-Ile86-Ser87 was positioned around P1 site of AKTI1. Sequence analysis suggested that two forms (single-chain and two-chain form) of AKPI2 protein were present in the tubers. Recombinant AKPI2 expressed by E.coli system showed inhibitory activity toward serine proteases and heat stability. The Ki values toward chymotrypsin and trypsin were 4 × 10-7 M and 6 × 10-6 M, respectively.Abbreviations: AAL: Apios americana lectin; AATI: Apios americana Bowman-Birk type trypsin inhibitor; ACE: angiotensin-converting enzyme; IPTG: isopropyl-β-D-thio-galactopyranoside; Ki: inhibition constant; KPIs: Kunitz-type protease inhibitors; L-BAPA: Benzoyl-L-arginine p-nitroanilide monohydrochloride; L-BTPA: Benzoyl-L-tyrosine p-nitroanilide; PFLNA: Pyr-Phe-Leu-p-nitroanilide; RP-HPLC: reverse-phase high-performance liquid chromatography; RT-PCR: reverse transcription-polymerase chain reaction; SDS-PAGE: sodium dodecyl sulfate-polyacrylamide gel electrophoresis; SLIC: sequence and ligation independent cloning; STANA: N-Succinyl-Ala-Ala-Ala-p-nitroanilide; SHR: spontaneously hypertensive rats; TFA: trifluoroacetic acid; UTR: untranslated region.

    更新日期:2020-01-10
  • Molecular insights into the mechanism of substrate recognition of Streptomyces transglutaminases.
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : null
    Shota Tokai,Misugi Uraji,Tadashi Hatanaka

    The microbial TGase from Streptomyces mobaraensis has used in various food industries. However, the detailed substrate specificities of TGases from the Streptomyces species toward the natural peptides remains to be unclear. In this study, we conducted the comparison of two different TGases from Streptomyces mobaranensis (SMTG) and Streptomyces cinnamoneus (SCTG). To clarify the region associated with the characteristics of enzymes, we constructed a chimeric enzyme of CM, of which is consisted of N-terminal half of SCTG and C-terminal half of SMTG. To reveal the differences in the substrate specificity between SCTG and SMTG toward natural peptides, we investigated the time dependence of TGase activity on the productivity of cross-linking peptide with tryptic casein and lysine by using LC-MS. We identified two peptides of "VLPVPQK" and "AVPYPQR" as substrates for both of the TGases.

    更新日期:2020-01-10
  • Changes in the physicochemical properties of fish cell membranes during cellular senescence.
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : null
    Akifumi Shiomi,Kohjiro Nagao,Hisae Kasai,Yuji Hara,Masato Umeda

    Fish cell lines are widely used for the studies of developmental biology, virology, biology of aging, and nutrition physiology. However, little is known about their physicochemical properties. Here, we report the phospholipid compositions and mechanical properties of cell membranes derived from freshwater, anadromous and marine fish species. Biophysical analyses revealed that fish cell lines have highly deformable cell membranes with significantly low membrane tensions and Young's moduli compared with those of mammalian cell lines. The induction of cellular senescence by DNA demethylation using 5-Aza-2'-deoxycytidine significantly reduced the deformability of fish cell membrane, but hydrogen peroxide-induced oxidative stress did not affect the deformability. Mass spectrometry analysis of phospholipids revealed that the level of phosphatidylethanolamine molecules containing polyunsaturated fatty acids significantly increased during the 5-Aza-2'-deoxycytidine-induced cellular senescence. Fish cell lines provide a useful model system for studying the changes in the physicochemical properties of cell membranes during cellular senescence.Abbreviations: 2D-TLC: two-dimensional thin layer chromatography; 5-Aza-dC: 5-Aza-2'-deoxycytidine; DHA: docosahexaenoic acid; EPA: eicosapentaenoic acid; FBS: fetal bovine serum; PC: phosphatidylcholine; PE: phosphatidylethanolamine; PI: phosphatidylinositol; PS: phosphatidylserine; PUFA: polyunsaturated fatty acid; SA-β-gal: senescence-associated beta-galactosidase; SM: sphingomyelin.

    更新日期:2020-01-10
  • Identification of Reg3β-producing cells using IL-22-stimulated enteroids.
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : null
    Mika Sato,Akihiko Inaba,Ken Iwatsuki,Yuki Saito,Miki Tadaishi,Makoto Shimizu,Kazuo Kobayashi-Hattori

    Reg3β, a lectin, displays antibacterial activity. This study investigated Reg3β-expressing cells using IL-22-stimulated enteroids. IL-22 stimulation elevated the mRNA and protein levels of Reg3β. IL-22 also increased the mRNA levels of CD133 (a transit-amplifying cell marker) and lysozyme (a Paneth cell marker). Immunohistochemistry showed partial colocalization of Reg3β- and lysozyme-positive cells, suggesting that Paneth cells are one of Reg3β-producing cells.

    更新日期:2020-01-10
  • Antidiabetic components from the hexane extract of red kidney beans (Phaseolus vulgaris L.): isolation and structure determination.
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : null
    Anita Maya Sutedja,Emiko Yanase,Irmanida Batubara,Dedi Fardiaz,Hanifah Nuryani Lioe

    Red kidney beans (Phaseolus vulgaris L.) contain bioactive compounds that are known to exhibit antidiabetic effects via inhibition of α-glucosidase. However, information on the nonpolar components that exhibit antidiabetic activity is limited. Here, we report the isolation and structure determination of components with α-glucosidase inhibitory activity, which were obtained from the hexane extract of red kidney beans. Triacylglycerols (TAGs) were identified as the major components exhibiting inhibitory activity against α-glucosidase. The chemical structure of TAGs was determined by a combination of GC-MS and UPLC-MS/MS. The primary TAGs identified were LnLnLn (trilinolenin) and LnLLn (1,3-dilinolenoyl-2-linoleoyl glycerol). The major fatty acids present in these TAGs were α-linolenic acid (ω-3) and linoleic acid (ω-6). These TAGs were also found to inhibit the α-glucosidase activity in a similar fashion as acarbose. These results suggest that TAGs have potency as antidiabetics and support the potential suitability of red kidney beans for diabetes treatment.

    更新日期:2020-01-10
  • Royal jelly reduces depression-like behavior through possible effects on adrenal steroidogenesis in a murine model of unpredictable chronic mild stress.
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : null
    Noriyuki Iegaki,Yukio Narita,Noriko Hattori,Yoko Hirata,Kenji Ichihara

    Royal jelly (RJ) is used as a dietary supplement for human health promotion. Recently, a clinical trial has reported that RJ improved mental health. The present study was conducted to experimentally support the clinical effect of RJ on mental health and to further elucidate the mechanisms of action of RJ. RJ and an ethanol extract of RJ, which contains fatty acids but not proteins, inhibited an unpredictable chronic mild stress (UCMS)-induced increase in immobility time, a depression-like behavior, in the tail suspension test. DNA microarray analysis of the adrenal grand revealed that the expression of genes involved in cholesterol metabolism was up-regulated in response to UCMS exposure and that RJ suppressed expression of genes related to cholesterol synthesis and transport. These results suggested that RJ improves stress-induced depression-like behavior by regulating adrenal steroidogenesis and that fatty acids contained in RJ partly contribute to the antidepressant effect of RJ.

    更新日期:2020-01-10
  • Effect of dietary cellulose nanofiber and exercise on obesity and gut microbiota in mice fed a high-fat-diet.
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : null
    Takao Nagano,Hiromi Yano

    Cellulose nanofiber (CN) consumption with exercise could be a potential strategy to control obesity. Here, we studied the effects of CN supplementation and voluntary exercise on obesity and gut microbiota in high-fat diet (HFD)-fed mice. Consumption of CN increased voluntary wheel running activity. CN intake and exercise together suppressed the increase in body weight and fat mass, and improved glucose tolerance. The fecal gut microbiota was analyzed by sequencing 16S ribosomal RNA genes. Principal component analysis revealed a shift in the microbiota composition resulting from exercise, but not from CN supplementation. Erysipelotrichaceae and Rikenellaceae decreased with exercise. Exercise also increased Ruminococcaceae, whereas exercise and CN intake together increased Eubacteriaceae. These two families are butyrate producers. Exercise increased the amount of acetate in the cecum. These results suggest that CN consumption improves exercise performance and exerts anti-obesity effects by modulating the balance of the gut microbiota.

    更新日期:2020-01-10
  • In vitro and in vivo evaluation of antioxidant activity of Petasites japonicus Maxim. flower buds extracts.
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : null
    Miki Hiemori-Kondo,Mika Nii

    The antioxidant activity of Petasites japonicus flower buds cultivated in Tokushima, Japan, was examined in vitro and in vivo. The flower bud extracts were assayed using either oxygen radical absorbance capacity or 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity. Antioxidants in the 80% ethanol extract were investigated using online high-performance liquid chromatography-DPPH and were identified as caffeic acid, 3-O-caffeoylquinic acid, fukinolic acid, 3,4-di-O-caffeoylquinic acid, 3,5-di-O-caffeoylquinic acid, and 4,5-di-O-caffeoylquinic acid using liquid chromatography-mass spectrometry. Fukinolic acid was the most active compound based on its activity and abundance. Administering the extracts orally to ICR mice prior to iron injection significantly suppressed plasma thiobarbituric acid reactive substance (TBARS) production. Moreover, TBARS and triglyceride concentrations in the plasma of C57BL/6 mice fed with a high fat diet were also significantly decreased by the extract. The results suggest that antioxidative compounds in P. japonicus can be used in the management of oxidative stress.

    更新日期:2020-01-10
  • Growth of term infants fed a commercial infant formula with a protein content of 2.2 g/100 kcal: an observational follow-up study.
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : null
    Shinji Jinno,Kae Yamazaki,Yoshitaka Nakamura,Toshi Kinouchi

    To evaluate the suitability of the new nutritional composition of renewed commercial Formula A (protein reduced to 2.2 g/100 kcal, arachidonic acid increased to 13.2 mg/100 kcal, and docosahexaenoic acid maintained at 20 mg/100 kcal), we examined whether the growth of Formula A-fed infants was equivalent to that of breastfed infants. In this observational study, 1,053 infants were followed-up to 12 months. Growth, stool consistency, and the health condition of 99 infants fed with Formula A and 295 breastfed infants were compared. Body weight, body mass index, and head circumference of Formula A-fed infants were similar to those of breastfed infants. Additionally, there were no differences in the stool consistency and the health condition (infection and allergy prevalence) between the two groups. Formula A-fed infants grew as well as breastfed infants, suggesting the appropriate nutritional composition of Formula A. The findings may contribute to further improvements in infant formulas.

    更新日期:2020-01-10
  • Isolation of four xylanases capable of hydrolyzing corn fiber xylan from Paenibacillus sp. H2C.
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : null
    Yusuke Hagiwara,Yasuhiro Mihara,Koichi Sakagami,Ryuta Sagara,Undramaa Bat-Erdene,Rie Yatsunami,Satoshi Nakamura

    Corn fibre xylan (CX) shows high resistance to enzymatic hydrolysis due to its densely decorated side chains. To find enzymes capable of hydrolyzing CX, we isolated a bacterial strain (named H2C) from soil, by enrichment culture using non-starch polysaccharides of corn as the sole carbon source. Analysis based on the 16S rRNA sequence placed strain H2C within genus Paenibacillus. Enzymes were purified from supernatant of culture broth of strain H2C based on solubilizing activities toward CX. Four enzymes, Xyn5A, Xyn10B, Xyn11A, and Xyn30A, were successfully identified, which belong to glycoside hydrolase (GH) families, 5, 10, 11, and 30, respectively. Phylogenetic analysis classified Xyn5A in subfamily 35 of GH family 5, a subfamily of unknown function. Their activities toward beechwood xylan and/or wheat arabinoxylan indicated that these enzymes are β-1,4-xylanases. They showed high solubilizing activities toward a feed material, corn dried distiller's grains with solubles, compared to five previously characterized xylanases.Abbreviations : CX: corn fibre xylan; DDGS: corn dried distiller's grains with solubles.

    更新日期:2020-01-10
  • Synthesis of nucleotide analogues, EFdA, EdA and EdAP, and the effect of EdAP on hepatitis B virus replication
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : 2019-10-07
    Mai Kamata; Toshifumi Takeuchi; Ei Hayashi; Kazane Nishioka; Mizuki Oshima; Masashi Iwamoto; Kota Nishiuchi; Shogo Kamo; Shusuke Tomoshige; Koichi Watashi; Shinji Kamisuki; Hiroshi Ohrui; Fumio Sugawara; Kouji Kuramochi

    4′-Ethynyl-2-fluoro-2′-deoxyadenosine (EFdA) and 4′-ethynyl-2′-deoxyadenosine (EdA) are nucleoside analogues which inhibit human immunodeficiency virus type 1 (HIV-1) reverse transcriptase. EdAP, a cyclosaligenyl (cycloSal) phosphate derivative of EdA, inhibits the replication of the influenza A virus. The common structural feature of these compounds is the ethynyl group at the 4′-position. In this study, these nucleoside analogues were prepared by a common synthetic strategy starting from the known 1,2-di-O-acetyl-D-ribofuranose. Biological evaluation of EdAP revealed that this compound reduced hepatitis B virus (HBV) replication dose-dependently without cytotoxicity against host cells tested in this study.

    更新日期:2019-12-19
  • Electrostatic interactions at the interface of two enzymes are essential for two-step alkane biosynthesis in cyanobacteria
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : 2019-10-10
    Mari Chang; Keigo Shimba; Yuuki Hayashi; Munehito Arai

    Cyanobacterial alkane biosynthesis is catalyzed by acyl-(acyl carrier protein (ACP)) reductase (AAR) and aldehyde-deformylating oxygenase (ADO) in a two-step reaction. AAR reduces acyl-ACPs to fatty aldehydes, which are then converted by ADO to alkanes, the main components of diesel fuel. Interaction between AAR and ADO allows AAR to efficiently deliver the aldehyde to ADO. However, this interaction is poorly understood. Here, using analytical size-exclusion chromatography (SEC), we show that electrostatic interactions play an important role in the binding of the two enzymes. Alanine-scanning mutagenesis at charged residues around the substrate entry site of ADO revealed that E201A mutation greatly reduced hydrocarbon production. SEC measurement of the mutant demonstrated that E201 of ADO is essential for the AAR–ADO interaction. Our results suggest that AAR binds to the substrate entrance gate of ADO and thereby facilitates the insertion of the reactive and relatively insoluble aldehyde into the hydrophobic channel of ADO. Abbreviations: AAR: acyl-ACP reductase; ACP: acyl carrier protein; ADO: aldehyde-deformylating oxygenase; ASA: solvent accessible surface area; BSA: bovine serum albumin; CD: circular dichroism; DMSO: dimethyl sulfoxide; DTT: dithiothreitol; GC-MS: gas chromatography-mass spectrometer; HPLC: high-performance liquid chromatography; IPTG: isopropyl-β-D-thiogalactoside; MRE: mean residue ellipticity; NpAAR: AAR from Nostoc punctiforme PCC 73102; NpADO: ADO from Nostoc punctiforme PCC 73102; PmADO: ADO from Prochlorococcus marinus MIT 9313; SDS-PAGE: sodium dodecyl sulfate-polyacrylamide gel electrophoresis; SeAAR: AAR from Synechococcus elongatus PCC 7942; SeADO: ADO from Synechococcus elongatus PCC 7942; SEC: size-exclusion chromatography; TeAAR: AAR from Thermosynechococcus elongatus BP-1; TeADO: ADO from Thermosynechococcus elongatus BP-1; UV: ultraviolet

    更新日期:2019-12-19
  • Binding of cloned Cel enzymes on clay minerals related to the pI of the enzymes and database survey of cellulases of soil bacteria for pI
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : 2019-10-18
    Renukaradhya K. Math; Devaiah Kambiranda; Han Dae Yun; Yemane Ghebreiyessus

    The Cel genes from Bacillus licheniformis MSB03 were cloned and expressed to investigate binding ability on clay minerals and sea sand at pH ranging 3 to 9. FTIR analysis has been done to characterize bound enzymes on clay minerals. Subsequent, surveying of NCBI database for extracellular enzymes of soil bacteria was carried out. Among the five cloned Cel enzymes assayed for binding to clay minerals, only Cel5H enzyme had the binding ability. Enzyme Cel5H exhibited highest binding to montmorillonite followed by kaolinite and sea sand. Interestingly, Cel5H had higher pI value of 9.24 than other proteins (5.2–5.7). Cel5H binding to montmorillonite was shown to be negatively affected below pH 3 and above pH 9. Infrared absorption spectra of the Cel5H-montmorillonite complexes showed distinct peaks for clay minerals and bound proteins. Furthermore, database survey of soil bacterial extracellular enzymes revealed that Bacillus species enzymes had higher pI than other soil bacterial enzymes.

    更新日期:2019-12-19
  • Cl- channels regulate lipid droplet formation via Rab8a expression during adipocyte differentiation
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : 2019-10-10
    Kanae Ouchi; Susumu Yoshie; Masao Miyake; Akihiro Hazama

    Several studies have shown that Cl− channels regulate the differentiation of some cell types. Thus, we investigated the role of Cl− channels on adipocyte differentiation using adipose tissue-derived stem cells (ASCs) and Cl− channel blocker. We induced rabbit ASCs into adipocytes using Cl− channel blocker. The expression levels of adipocyte markers were no significant difference between the cells treated with a Cl− channel blocker NPPB and untreated cells. However, when the cells were treated with NPPB, lipid droplets (LDs) sizes decreased compared with the untreated control. Interestingly, the expression levels of Rab8a, which is known as a regulator of LD fusion, were also decreased in the cells treated with NPPB. Other Cl− channel blockers, DIDS and IAA-94, also inhibited large LDs formation and Rab8a expression. These results demonstrate that Cl− channels do not regulate the adipocyte differentiation, but do regulate the LDs formation via Rab8a expression. Abbreviations: ASCs: adipose tissue-derived stem cells; LDs: lipid droplets; RUNX2: runt-related transcription factor 2; CFTR: cystic fibrosis transmembrane conductance regulator; TG: triacylglycerol; FA: fatty acid; GLUT4: glucose transporter type 4; ER: endoplasmic reticulum; ADRP: adipose differentiation-related protein; TIP47: tail-interacting protein of 47 kD; HSL: hormone sensitive lipase; PBS: phosphate-buffered saline; DMEM: Dulbecco’s modified Eagle Medium; FBS: fetal bovine serum; SMA: smooth muscle actin; FAS: fatty acid synthase; ZONAB: ZO-1 associated nucleic acid binding protein; PPAR-γ: peroxisome proliferator-activated receptor-γ; C/EBPα: CCAAT/enhancer binding protein α; CE: cholesteryl ester; V-ATPase: vacuolar H+ ATPase.

    更新日期:2019-12-19
  • Mesenchymal stem cells co-cultured with colorectal cancer cells showed increased invasive and proliferative abilities due to its altered p53/TGF-β1 levels
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : 2019-10-10
    In-Rok Oh; Bernardo Raymundo; MiJung Kim; Chan-Wha Kim

    Signaling between cancer cells, their neighboring cells, and mesenchymal stem cells (MSCs) forms the tumor microenvironment. The complex heterogeneity of this microenvironment varies depending on the tumor type and its origins. However, most of the existing cancer-based studies have focused on cancer cells. In this study, we used a direct co-culture system (cross-talk signaling) to induce cross-interaction between cancer cells and mesenchymal stem cells. This induced deformation of MSCs. MSCs showed a diminished ability to maintain homeostasis. In particular, increase in the invasion ability of MSCs by TGF-β1 and decrease in p53, which plays a key role in cancer development, is an important discovery. It can thus be deduced that blocking these changes can effectively inhibit metastatic colorectal cancer. In conclusion, understanding the interactions and changes in MSCs associated with cancer will help develop novel therapeutic strategies for cancer.

    更新日期:2019-12-19
  • Cryptotanshinone attenuates allergic airway inflammation through negative regulation of NF-κB and p38 MAPK
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : 2019-11-05
    Junfeng Li; Mingyu Zheng; Chongyang Wang; Jingzhi Jiang; Chang Xu; Li Li; Liangchang Li; Guanghai Yan; Yongde Jin

    This study is to determine the role and mechanism of cryptotanshinone (CTS) in allergic airway inflammation. Asthma induced by OVA was established in BALB/c mice. We found increased airway hyperresponsiveness (AHR), increased inflammatory cell infiltration, elevated levels of TNF-α, interleukin-1β (IL-1β), IL-4, IL-5, IL-6 and IL-13, decreased interferon gamma (IFN-γ) in lung tissue, increased content of total immunoglobulin E (IgE), OVA specific IgE, Eotaxin, ICAM-1, VCAM-1, nuclear factor-kappaB (NF-κB) and phosphorylation of p38 MAPK in lung tissue. However, the administration of CTS significantly decreased AHR in asthmatic mice, reduced inflammation around the bronchioles and inflammatory cells around airway, regulated cytokine production, reduced the total IgE and OVA-specific IgE levels, and inhibited NF-κB activation and p38 MAPK phosphorylation. In vitro experiments in 16 HBE cells revealed that CTS attenuated CAM-1 and IL-6 expression. These results indicate that CTS alleviates allergic airway inflammation by modulating p38 MAPK phosphorylation and NF-κB activation.

    更新日期:2019-12-19
  • GRM4 inhibits the proliferation, migration, and invasion of human osteosarcoma cells through interaction with CBX4
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : 2019-10-04
    Zengliang Zhang; Nan Li; Xing Wei; Bingyao Chen; Yinglong Zhang; Yantao Zhao; Xiantong Hu; Shuxun Hou

    In recent years, the survey of metabolic glutamate receptor 4 (GRM4) in tumor biology has been gradually concerned. There are currently few studies on GRM4 in osteosarcoma, and the biological function is not clear. Analysis of TCGA database showed that there was no substantial deviation in the expression of GRM4 between osteosarcoma and normal tissues. In the subsequent experiments, there is no significant difference in either mRNA or protein levels among immortalized human osteoblasts and various osteosarcoma cells. With the overexpression of GRM4, cell proliferation, migration and invasion were inhibited obviously. It was further revealed that GRM4 can interact with CBX4 to restrict the nuclear localization of CBX4 and affect the transcriptional activity of HIF-1α. This is the evidence supporting the interaction between GRM4 and CBX4, which could inhibit the malignant behavior of osteosarcoma cells through the GRM4/CBX4/HIF-1α signaling pathway.

    更新日期:2019-12-19
  • Leptomycin B inhibits the proliferation, migration, and invasion of cultured gastric carcinoma cells
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : 2019-10-16
    Hepan Zhu; Yi Yang; Li Wang; Xiaobin Xu; Tingting Wang; Haoran Qian

    Chromosome region maintenance 1 (CRM1) plays a critical role in tumorigenesis and progression through modulating nuclear export of several proteins. However, the precise effects of CRM1 inhibitor on gastric carcinoma have not yet been illustrated. Here, we investigated the potential anti-cancer activities of leptomycin B, the most potent CRM1 antagonist, on cultured gastric carcinoma cells. Our findings demonstrate that CRM1 was highly expressed in four gastric carcinoma cell lines. Leptomycin B inhibited the viability of HGC-27 and AGS cells in a dose- and time-dependent pattern. Leptomycin B at the dose of 10 nM or 100 nM suppressed the migration and invasion of HGC-27 and AGS cells. Leptomycin B elevated the expressions of autophagy-related protein LC3-II and autophagy substrate p62. Moreover, leptomycin B enhanced the LC3-positive puncta formation in cells. Our data suggest that leptomycin B may exert an anti-cancer activity possibly through interfering autophagy function in gastric carcinoma cells.

    更新日期:2019-12-19
  • MicroRNA-519d-3p inhibits cell proliferation and cell cycle G1/S transition in glioma by targeting CCND1
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : 2019-10-29
    Lishan Ma; Jin Li

    Glioma is the most common highly malignant primary brain tumor. MicroRNA-519d-3p exerts important effects in several tumors, but its functional role in glioma remained poorly understood. In this study, we found miR-519d-3p expression was significantly decreased in glioma tissues and cell lines. Moreover, the in vitro experiments showed that overexpression of miR-519d-3p suppressed cell proliferation and induced cell cycle G0/G1 phase arrest using MTT and flow cytometry assays in glioma cell lines, U87 and U251. Mechanistically, Cyclin D1 (CCND1) was predicted and confirmed as the direct target genes of miR-519d-3p using luciferase report assay. In addition, knockdown of CCND1 imitated the suppressive effects of miR-519d-3p on cell proliferation and cell cycle progression. Furthermore, restoration of CCND1 reversed the effects of miR-519d-3p overexpression in glioma cells. Taken together, these data demonstrate that suppression of CCND1 by miR-519d-3p might be a therapeutic target for glioma. Abbreviations miR-519d-3p: microRNA-519d-3p; CCND1: Cyclin D1; ATCC: American Type Culture Collection; MTT: 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide; PI: propidium iodide; WT: wild type; MUT: mutant type; SD: standard deviation

    更新日期:2019-12-19
  • Long non-coding RNA 2310069B03Rik functions as a suppressor of Ucp1 expression under prolonged cold exposure in murine beige adipocytes
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : 2019-10-10
    Mari Iwase; Shoko Sakai; Shigeto Seno; Yu-Sheng Yeh; Tony Kuo; Haruya Takahashi; Wataru Nomura; Huei-Fen Jheng; Paul Horton; Naoki Osato; Hideo Matsuda; Kazuo Inoue; Teruo Kawada; Tsuyoshi Goto

    Specific conditions, such as exposure to cold, can induce the production of brown-like adipocytes in white adipose tissue. These adipocytes express high levels of uncoupling protein 1 (UCP1) and energy expended by generating heat. Thus, these are a potential target for the prevention or treatment of obesity. The present study involved a comprehensive analysis of the adipose tissue to understand the relationship between long non-coding RNA (lncRNA) 2310069B03Rik and UCP1. Cold exposure increased both lncRNA 2310069B03Rik and Ucp1 expression in inguinal white adipose tissue (iWAT). However, overexpression of lncRNA 2310069B03Rik suppressed the Ucp1 mRNA expression and the promoter activity of UCP1 in the iWAT primary adipocytes. In addition, compared to the early induction of Ucp1 expression by cold stimulation, the induction of lncRNA 2310069B03Rik expression was later. These results suggest that lncRNA 2310069B03Rik functions as a suppression factor of Ucp1 expression.

    更新日期:2019-12-19
  • Glycyrrhiza glabra suppresses nasopharyngeal carcinoma cell proliferation through inhibiting the expression of lncRNA, AK027294
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : 2019-10-07
    Bo Zhang; Min Yan; Wei Zhang; Zhao-yang Ke; Ling-guo Ma

    Glycyrrhiza glabra is considered as potential drug for nasopharyngeal carcinoma (NPC). However, whether the long noncoding RNAs’ (lncRNAs) contributes to the anti-cancer function of this herb is unknown. In present study, we analyzed the differential expression of lncRNA between G. glabra-treated and untreated C666-1 cells. Out of those tumor-related lncRNAs, AK027294 had a strongest down-regulation upon G. glabra treatment. Knockdown of AK027294 suppresses the proliferation of C666-1 cells by inducing the apoptosis. Moreover, either G. glabra treatment or knockdown of AK027294 significantly increases the production of EZH1 (Enhancer of zeste 1 polycomb repressive complex 2 subunit). Collectively, we have identified a potential mechanism that the down-regulation of AK027294 contributes to the anti-cancer function of G. glabra and also provide the potential inter-relationship between AK027294 and EZH1.

    更新日期:2019-12-19
  • MiR-210-3p attenuates lipid accumulation and inflammation in atherosclerosis by repressing IGF2
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : 2019-11-02
    Xiang-Rui Qiao; Liang Wang; Mengping Liu; Yuling Tian; Tao Chen

    Previous studies have shown that miR-210-3p is involved in the development and progression of atherosclerosis, but its specific mechanisms are still unclear. This study aims to reveal the mechanism of miR-210-3p and its target genes in macrophage lipid deposition and inflammatory response, and provide new ideas for the treatment of atherosclerosis. We found miR-210-3p increased sharply in the first 12 h induced by higher doses of ox-LDL in THP-1 macrophages and then gradually decreased. MiR-210-3p mimic transfection inhibited lipid uptake and inflammatory cytokine production in ox-LDL-induced macrophages. By inhibiting IGF2/IGF2R, miR-210-3p suppressed the expression of fatty acid transcriptase CD36 and transcription factor NF-κB in ox-LDL-induced macrophages. In conclusion, miR-210-3p inhibits the expression of CD36 and NF-κB by inhibiting IGF2 / IGF2R, thereby reducing lipid accumulation and inflammatory response in ox-LDL-induced macrophages. Enhancing miR-210-3p expression may be a new strategy for the treatment of atherosclerosis.

    更新日期:2019-12-19
  • gas1 mutation extends chronological lifespan via Pmk1 and Sty1 MAPKs in Schizosaccharomyces pombe
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : 2019-10-10
    Yuki Imai; Takafumi Shimasaki; Chihiro Enokimura; Hokuto Ohtsuka; Satoshi Tsubouchi; Kunio Ihara; Hirofumi Aiba

    In the longevity research by using yeasts, chronological lifespan is defined as the survival time after entry into stationary phase. Previously, screening for long lived mutants of Schizosaccharomyces pombe was performed to identify the novel factors involved in longevity. From this screening, one long lived mutant called as No.36 was obtained. In this study, we identified the mutation caused in gas1+, which encodes glucanosyltransferase (gas1-287 mutation) is responsible for the longevity of No.36 mutant. Through the analysis of this mutant, we found that cell wall perturbing agent micafungin also extends chronological lifespan in fission yeast. This lifespan extension depended on both Pmk1 and Sty1 MAP kinases, and longevity caused by the gas1-287 mutation also depended on these kinases. In summary, we propose that the gas1-287 mutation causes longevity as the similar mechanism as cell wall stress depending on Pmk1 and Sty1 MAPK pathways.

    更新日期:2019-12-19
  • MicroRNA-4461 derived from bone marrow mesenchymal stem cell exosomes inhibits tumorigenesis by downregulating COPB2 expression in colorectal cancer
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : 2019-10-21
    Hui-Li Chen; Jiu-Jiang Li; Fei Jiang; Wen-Jing Shi; Ge-Yun Chang

    Colorectal cancer (CRC) is one of the main cause of cancer-related deaths. It’s reported that bone marrow mesenchymal stem cells (BMSCs) affects tumor development through secreting exosomes. This study aims to investigate the function of BMSCs-derived exosome miR-4461 in CRC. The results of qRT-PCR showed that miR-4461 expression in DLD1, HCT116 and SW480 CRC cells and CRC tissues was lower than that in FHC cells and normal tissues, respectively. And COPB2 mRNA expression was negatively correlated with miR-4461. Western blot was used to detect COPB2 protein expression. Dual-luciferase reporter assay results revealed that miR-4461 targeted COPB2. Transwell assay and CCK-8 assay demonstrated that COPB2 knockdown inhibited HCT116 and SW480 cells proliferation, migration and invasion abilities. Furthermore, BMSCs-derived exosome miR-4461 downregulated COPB2 expression and inhibited HCT116 and SW480 cells migration and invasion. The findings demonstrated that miR-4461 could be a potential target for the diagnosis and treatment of colorectal cancer.

    更新日期:2019-12-19
  • Identification of critical residues for the catalytic activity of ComQ, a Bacillus prenylation enzyme for quorum sensing, by using a simple bioassay system
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : 2019-10-31
    Kazutake Hirooka; Saki Shioda; Masahiro Okada

    Bacillus ComQ participates in the biosynthesis of a quorum-sensing signaling molecule (ComX pheromone) through catalyzing the prenylation at a Trp residue of the precursor peptide (pre-ComX) with geranyl diphosphate (C10 type) or farnesyl diphosphate (C15 type). We hypothesized that several residues specifically conserved among either type of ComQs are important for their substrate specificities. Using a simple bioassay, we revealed that Phe63, Asn186, and Gly190 in ComQRO-E-2 (C10 type) were nondisplaceable to Ser63, Gly186, and Val190, the corresponding residues in the C15-type ComQ, respectively. A three-dimensional model suggested that the 186th and 190th residues are involved in the pre-ComX binding. In vitro analysis showed that substitution of Phe63 with Ser in ComQRO-E-2 significantly reduced the geranylation activity but substantially enhanced the farnesylation activity, whereas substitution of Ser63 with Phe in ComQ168 (C15 type) reduced the farnesylation activity. Therefore, the 63rd residue was found to be significant for the prenyl-substrate preference. Abbreviations: GPP: geranyl diphosphate; FPP: farnesyl diphosphate; IPP: isopentenyl diphosphate; GGPP: geranylgeranyl diphosphate; FARM: first aspartate-rich motif; SARM: second aspartate-rich motif; β-Gal: β-galactosidase; TBABG: tryptose blood agar base supplemented with glucose; X-gal: 5-bromo-4-chloro-3-indolyl-β-D-galactoside

    更新日期:2019-12-19
  • Structural and functional evaluation of single-chain Fv antibody HyC1 recognizing the residual native structure of hen egg lysozyme
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : 2019-10-29
    Takanori Yamaoka; Yuji O. Kamatari; Takahiro Maruno; Yuji Kobayashi; Masayuki Oda

    Evaluation of the molecular mechanisms by which an antibody recognizes a specific antigen could help in better understanding of the protein recognition mechanisms. We previously showed that anti-hen egg lysozyme (HEL) monoclonal antibody, HyC1, recognized the structural and hydrodynamic change in HEL. Here, we generated HyC1 single-chain Fv (scFv), and characterized it using different structural and biophysical methods. Similar to HyC1 monoclonal antibody, HyC1 scFv could recognize native HEL from carboxymethylated Cys6 and Cys127 HEL (CM6,127-HEL). Comparison of the binding thermodynamics of HyC1 scFv between HEL and CM6,127-HEL showed that the binding enthalpy change was different, while the binding entropy was remained unchanged. The results indicated that the fluctuation of the residual native structure in both HEL and CM6,127-HEL was similar. The NMR experiments for 15N-labeled HyC1 scFv indicated that the flexibility of HyC1 scFv decreased upon the binding to HEL.

    更新日期:2019-12-19
  • Effects of single ingestion of rice cracker and cooked rice with high resistant starch on postprandial glucose and insulin responses in healthy adults: two randomized, single-blind, cross-over trials
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : 2019-11-05
    Yuhi Saito; Toshiyuki Watanabe; Toshiyuki Sasaki; Kenichi Watanabe; Masao Hirayama; Naoko Fujita

    Rice with double mutation of starch synthase IIIa and branching enzyme IIb (ss3a/be2b) has much higher amounts of apparent amylose and resistant starch (RS) than usual varieties. In this study, we conducted two randomized, single-blind, crossover trials to investigate the effect of single ingestion of two processed foods composed of ss3a/be2b mutant rice on postprandial blood glucose and insulin response in healthy adults, compared to those of usual cultivar. In trial 1, of ingestion of rice crackers, the incremental area under the curves of glucose (IAUCglc) and insulin (IAUCins) in RS group was significantly lower than in the control group. In trial 2, of the ingestion of cooked rice, IAUCglc in the RS group was significantly lower than in the control group. These results showed that the ss3a/be2b-mutant rice cracker and cooked rice having high RS can attenuate postprandial blood glucose and insulin response.

    更新日期:2019-12-19
  • Silk sericin intake leads to increases in L-serine and L-tyrosine levels in the mouse brain and the simultaneous facilitation of brain noradrenergic turnover
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : 2019-10-12
    Yui Kawano; Hu Yating; Masahiro Sasaki; Shigeki Furuya

    Sericin is a protein component of the silkworm cocoon, and contains a high proportion of L-serine, but it has been mostly disposed of as an industrial waste. However, recent studies have revealed its unique biological functionalities beneficial to human health. This study aimed to evaluate the effect of acute oral intake of sericin on amino acid and neurotransmitter metabolism in the mouse brain. Acute administration of chemically modified sericin (0.26 g/30 g body weight) increased L-serine and L-tyrosine levels in the serum and brain, although the L-tyrosine content in the sericin was less than 3% (w/w). In addition, sericin administration led to a significant facilitation of noradrenergic turnover via enhancement of 3-methoxy-4-hydroxyphenylethyleneglycol, a principal metabolite of noradrenaline, in several of the brain regions examined. These present findings suggest that oral intake of sericin efficiently delivers L-serine and L-tyrosine to the brain, thus stimulating noradrenergic activity in the brain. Abbreviations: DA: dopamine; 5-HIAA: 5-hydroxyindoleicetic acid; 5-HT: 5-hydroxytryptamine; HVA: homovanillic acid; MHPG: 3-methoxy-4-hydroxyphenylethyleneglycol; 3-MT: 3-methoxytyramine; NA: noradrenaline; NM: normetanephrine; Veh: vehicle

    更新日期:2019-12-19
  • Preparation and effects on neuronal nutrition of plasmenylethonoamine and plasmanylcholine from the mussel Mytilus edulis
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : 2019-10-12
    Yi Ding; Rui Wang; Xiaoxu Wang; Peixu Cong; Yanjun Liu; Zhaojie Li; Jie Xu; Changhu Xue

    Plasmenylethonoamine (pPE) and plasmanylcholine (aPC) are important phospholipid subclasses. Herein we explored optimum conditions for enzymatic purification and preparation of pPE and aPC from the mussel Mytilus edulis and bovine brain. Among them, pPE in Mytilus edulis PE was mainly p18:0–20:5 and p18:0–22:6, and its purity was 92.7%; aPC in PC was primarily a16:0–22:6 and a16:0–20:5, and aPC accounted for 90.2% of PC. We thereafter evaluated neurotrophic effects of Mytilus edulis pPE, aPC, and bovine brain pPE in a NGF-induced PC12 cell model. Morphologically, pPE and aPC could both promote differentiation, manifested in a significant increase in neurite length and number, due to increased expression of synaptophysin and growth protein GAP-43 in a dose-independent and structure-selective manner. Importantly, the effect on neuronal nutrition of pPE was better than aPC, and marine pPE was better than terrestrial pPE, which might be ascribed to vinyl-ether bond and differences in fatty acid composition. Abbreviations: AA: arachidonic acid; DHA: docosahexaenoic acid; EIC: extracted ion chromatogram; EPA: eicosapentanoic acid; GAP: growth-associated protein; HPLC: high-performance liquid chromatography; LC-MS/MS: liquid chromatography-tandem mass spectrometry; LPC: lyso-PC; LPE: lyso-PE; MS: mass spectrometry; NGF: nerve growth factor; PC: phosphatidylcholine; aPC: plasmanylcholine; PE: phosphatidylethanolamine; pPE: plasmenylethonoamine; PG: phosphoglycerols; PLs: phospholipids; PS: phosphoserines; TIC: total ion chromatogram.

    更新日期:2019-12-19
  • Maltobionic acid accelerates recovery from iron deficiency-induced anemia in rats
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : 2019-10-12
    Daiki Suehiro; Haruka Kawase; Sumire Uehara; Rio Kawase; Ken Fukami; Tomoyuki Nakagawa; Masaya Shimada; Takashi Hayakawa

    In experiments 1 and 2, effect of ingestion of maltobionic acid calcium salt (MBCa) on recovery of rats from a latent iron deficiency and from iron deficiency anemia was examined, respectively. After grouping rats into control and iron-deficiency groups, a latent iron deficiency or iron-deficiency anemia was induced in the latter group. And recovery from these states by MBCa containing diets (0%, 3%, and 6% MBCa in diet, classified into MBCa-0, MBCa-3, and MBCa-6 groups) was compared for convalescence period in light of iron sufficient control group. In experiment 1, MBCa ingestion significantly increased the iron concentration in the serum and liver, and promoted recovery from a latent iron deficiency. In experiment 2, hemoglobin and hematocrit levels increased significantly with MBCa intake, and recovery from iron-deficiency anemia was promoted. MBCa effectively promoted the recovery of rats from a subclinical iron deficiency and iron-deficiency anemia. Abbreviations: ANOVA: analysis of variance; DMT1: divalent metal transporter 1; EDTA-2Na: disodium salt of ethylenediaminetetraacetic acid; Fpn: feroportin; Hb: hemoglobin; Ht: hematocrit; ICP-OES: inductivity coupled plasma optical emission spectrometer; MBCa: maltobionic acid calcium salt; nitroso-PSAP: 2-nitroso-5-[N-n-propyl-N-(3-sulfopropyl)amino]phenol; SE: standard error; SI: serum-iron concentration; TSAT: transferrin saturation; TIBC: total iron-binding capacity; UIBC: unsaturated iron-binding capacity.

    更新日期:2019-12-19
  • Chamazulene reverses osteoarthritic inflammation through regulation of matrix metalloproteinases (MMPs) and NF-kβ pathway in in-vitro and in-vivo models
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : 2019-10-23
    Ding Ma; Jinlong He; Dapeng He

    This study was conducted to evaluate the protective effects of chamazulene against IL-1β-induced rat primary chondrocytes and complete Freund’s adjuvant (CFA)-induced osteoarthritic inflammation in rats. Oxidative stress markers, pro-inflammatory cytokines, and regulatory proteins were measured. Chamazulene significantly reverted (p < 0.05) the levels of lipid peroxidation and enhanced the activities of superoxide dismutase (SOD), glutathione peroxidase (GPx) and catalase (CAT) enzymes against IL-1β and CFA-induced oxidative stress. The levels of TNF-α and IL-6 were reduced (p < 0.05) in chamazulene treatment against IL-1β and CFA-induced inflammation. Western blot analysis results on the expressions of MMP-3, MMP-9, p65 NF-kβ, iNOS, and COX-2 showed chamazulene was able to protect the chondrocytes against IL-1β-induced osteoarthritic inflammation. Histopathology of rat hind ankle showed chamazulene significantly protected against CFA-induced osteoarthritic inflammation. Therefore, chamazulene can be recommended as a therapeutic agent for clinical trials against osteoarthritic inflammation.

    更新日期:2019-12-19
  • Antibiotics can cause weight loss by impairing gut microbiota in mice and the potent benefits of lactobacilli
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : 2019-10-29
    Zhonghua Miao; Ruyue Cheng; Yujie Zhang; Huijing Liang; Fengling Jiang; Xi Shen; Gong Chen; Qisheng Zhang; Fang He; Ming Li

    This study assessed whether antibiotics could alter gut microbiota to affect host growth and the possibility of alleviation by lactobacilli. We divided four-week-old BABL/c mice into control (Ctrl), antibiotic exposure (Abx), Lactobacillus plantarum PC-170 (PC), and Lactobacillus rhamnosus GG (LGG) group and the Abx, LGG, and PC group received an one-week antibiotic/antibiotic + probiotic treatment. The fecal microbiota and the expression of splenic cytokines were determined. Following the ceftriaxone treatment, the body weight gain of Abx was delayed compared with others. The ceftriaxone treatment significantly decreased the alpha-diversity of the fecal microbiota and altered the fecal microbiota but LGG and PC can partly alleviate the effect. At the end of the study, the microbial community of LGG and PC group were more similar to Ctrl compared with Abx group. The results indicated that ceftriaxone could significantly alter intestinal microbiota. Lactobacilli might alleviate the side effects of antibiotics by stabilizing the intestinal microbiota.

    更新日期:2019-12-19
  • Identification of the novel odor active compounds “p-menthane lactones” responsible for the characteristic aroma of fresh peppermint leaf
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : 2019-11-02
    Atsushi Shigeto; Atsushi Wada; Kenji Kumazawa

    The comparative aroma extract dilution analysis (cAEDA) applied to the volatile fractions of two types of fresh mint leaves (Peppermint and Spearmint) revealed 20 odor-active peaks with high FD factors (≥64). Among the perceived odorants, five “sweet/milky” and “caramel-like/spicy” notes showing high FD factors were characteristic for the peppermint aroma, and were identified as p-menthane lactone derivatives. Especially, the three p-menthane lactone derivatives, “rel-(3S,3aS,6R,7aS)-dihydromintlactone, (3S,3aS,6R)-epoxymenthofurolactone, and 2E-ethylidene-5-methylcyclohexanone”, were newly identified compounds in the peppermint. The biosynthetic routes of the p-menthane lactone derivatives have been suggested to be closely related to menthofuran. Therefore, these findings strongly suggest the possibility that the biosynthetic pathway leading to the p-menthane lactone derivatives via menthofuran plays a significantly important role in the formation of the characteristic aroma of peppermint.

    更新日期:2019-12-19
  • Z-Isomers of lycopene exhibit greater liver accumulation than the all-E-isomer in mice
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : 2019-10-12
    Masaki Honda; Yuki Nakayama; Sho Nishikawa; Takanori Tsuda

    The effect of oral administration of all-E-isomer-rich and Z-isomer-rich lycopene on liver accumulation in mice was investigated. When a diet rich in the Z-isomers was administered for 4 weeks, the total lycopene concentration in the liver was more than 3 times higher than that of all-E-isomer administration. This result clearly indicates that lycopene Z-isomers show greater bioavailability and/or liver accumulation than the all-E-isomer in mice.

    更新日期:2019-12-19
  • Effects of normothermic microwave irradiation on CD44+/CD24‒ in breast cancer MDA-MB-231 and MCF-7 cell lines
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : 2019-09-27
    Mamiko Asano; Satoshi Tanaka; Minoru Sakaguchi

    We previously reported that MDA-MB-231 and MCF-7 cells, which are breast cancer cell lines and have cancer and cancer-initiating cells (CICs), were killed following normothermic microwave irradiation in which the cellular temperature was maintained at 37°C. In this study, we investigated the percentages of live or dead cells among CD44+/CD24− cells, which were defined as CICs among MDA-MB-231 and MCF-7 cells, and other types of cells in response to microwave irradiation. CD44+/CD24− cells among MDA-MB-231 cells were killed, thereby decreasing the number of cells, whereas the number of live CD44+/CD24− MCF-7 cells was increased following microwave irradiation. Moreover, adhesion, invasion, and migration were decreased in MDA-MB-231 cells, and the activation of matrix metalloproteinase-2 (MMP-2) in MDA-MB-231 cells was increased following microwave irradiation. These decreased cell activities might have been caused by MMP-2 activation and population changes in CD44+/CD24− in MDA-MB-231 cells. Abbreviations: APC: allophecocyanin; CBB: coomassie Brilliant Blue; CD: cluster of differentiation; CICs: cancer-initiating cells; FACS: fluorescence-activated cell sorting; FBS: fetal bovine serum; FITC: fluorescein isothiocyanate; FTDT: finite-difference time domain; HER2: human epidermal growth factor receptor type 2; PI: propidium iodide.

    更新日期:2019-12-19
  • New diagnostic method for Alzheimer’s disease based on the toxic conformation theory of amyloid β
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : 2019-09-20
    Kazuhiro Irie

    Recent investigations suggest that soluble oligomeric amyloid β (Aβ) species may be involved in early onset of Alzheimer’s disease (AD). Using systematic proline replacement, solid-state NMR, and ESR, we identified a toxic turn at position 22 and 23 of Aβ42, the most potent neurotoxic Aβ species. Through radicalization, the toxic turn can induce formation of the C-terminal hydrophobic core to obtain putative Aβ42 dimers and trimers. Synthesized dimer and trimer models showed that the C-terminal hydrophobic core plays a critical role in the formation of high molecular weight oligomers with neurotoxicity. Accordingly, an anti-toxic turn antibody (24B3) that selectively recognizes a toxic dimer model of E22P-Aβ42 was developed. Sandwich enzyme-linked immunosorbent assay with 24B3 and 82E1 detected a significantly higher ratio of Aβ42 with a toxic turn to total Aβ42 in cerebrospinal fluid of AD patients compared with controls, suggesting that 24B3 could be useful for early onset of AD diagnosis.

    更新日期:2019-12-04
  • Large scale production of lacto-N-biose I, a building block of type I human milk oligosaccharides, using sugar phosphorylases
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : 2019-09-28
    Mamoru Nishimoto

    Human milk oligosaccharides (HMOs) have drawn attention for their contribution to the explosive bifidobacterial growth in the intestines of neonates. We found that bifidobacteria can efficiently metabolize lacto-N-biose I (LNB), the major building blocks of HMOs, and we have developed a method to synthesize LNB by applying this system. We produced LNB on a kilogram scale by the method. This proved that, among the enterobacteria, only bifidobacteria can assimilate LNB, and provided the data that supported the explosive growth of bifidobacteria in neonates. Furthermore, we were also able to reveal the structure of LNB crystal and the low stability for heating at neutral pH, which has not been clarified so far. In this paper, using bifidobacteria and LNB as examples, I describe the research on oligosaccharide synthesis that was conducted by utilizing a sugar metabolism. Abbreviations: LNB: lacto-N-biose I; GNB: galacto-N-biose; HMOs: human milk oligosaccharides; GLNBP: GNB/LNB phosphorylase; NahK: N-acetylhexosamine 1-kinase; GalT: UDP-glucose-hexose-1-phosphate uridylyltransferase; GalE: UDP-glucose 4-epimerase; SP: sucrose phosphorylase.

    更新日期:2019-12-04
  • Predominant accumulation of a 3-hydroxy-γ-decalactone in the male rectal gland complex of the Japanese orange fly, Bactrocera tsuneonis
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : 2019-09-13
    Hajime Ono, Masataka Nakahira, Satoshi Ohno, Jun Otake, Tomoya Kanno, Isao Tokushima, Yoshimitsu Higashiura, Ichiro Nishi, Ritsuo Nishida

    The Japanese orange fly, Bactrocera tsuneonis, infests various citrus crops. While male pheromone components accumulated in the rectal glands are well characterized for Bactrocera, but information regarding the chemical factors involved in the life cycles of B. tsuneonis remains scarce. Herein, several volatile chemicals including a γ-decalactone, (3R,4R)-3-hydroxy-4-decanolide [(3R,4R)-HD], were identified as major components, along with acetamide and spiroketals as minor components in the rectal gland complexes of male B. tsuneonis flies. The lactone (3R,4R)-HD was also identified in female rectal gland complexes. The amount of this compound in mature males was significantly higher than those observed in females and immature males. The lactone (3R,4R)-HD was detected in flies fed with sucrose only, indicating that this lactone is not derived from dietary sources during adulthood, but biosynthesized in vivo. The predominant accumulation of (3R,4R)-HD in mature males also suggests a possible role in reproductive behavior.

    更新日期:2019-12-04
  • Isolation, synthesis, and biological activities of a bibenzyl from Empetrum nigrum var. japonicum
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : 2019-09-06
    Sayuki Oka, Ryo Kuniba, Nozomi Tsuboi, Sayaka Tsuchida, Kazunari Ushida, Shusuke Tomoshige, Kouji Kuramochi

    4-(2-Hydroxyphenethyl)-2,6-dimethoxyphenol, a bibenzyl, was isolated from the leaves of Empetrum nigrum var. japonicum, collected from Mount Tateyama. Japanese rock ptarmigans frequently eat the leaves and fruits of this plant. The structure of the bibenzyl was confirmed by NMR spectroscopic analysis and fully characterized. A synthesis of this compound was accomplished by coupling 2-hydroxyphenylacetic acid with syringaldehyde, decarboxylation of the resultant isoaurones, and hydrogenation of the double bond in the corresponding stilbene. This compound displayed cytotoxic activity against human cancer cells (HCT116 and Hela cells) and leukemia cells (HL-60 cells). The present study suggests that this plant serves as a source of biologically active natural products. Also, our findings provide information on the secondary metabolites in the diet of Japanese rock ptarmigans.

    更新日期:2019-12-04
  • Asymmetric synthesis of trans-p-menth-3-ene-1,2,8-triol, the monoterpene isolated from herbal plants
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : 2019-10-03
    Shunsuke Konishi, Yusuke Ogura, Hirosato Takikawa, Hidenori Watanabe

    The monoterpene, trans-p-menth-3-ene-1,2,8-triol, is a naturally occurring alcohol isolated from several herbal plants. In the present work, the asymmetric synthesis of both enantiomers of this natural product was achieved using Sharpless asymmetric dihydroxylation as the key step. A reversal of enantiofacial selectivity was observed in the asymmetric dihydroxylation.

    更新日期:2019-12-04
  • Visual, sensitive and rapid event-specific detection of genetically modified potato EH92-527-1 by loop-mediated isothermal amplification method
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : 2019-09-07
    Yuan-Kai Tu, Yen-Chun Lin, Yu-Wei Feng, Yeu-Yang Tseng, Han-Wei Chen

    To date, studies on the application of loop-mediated isothermal amplification (LAMP) in the detection of genetically modified organisms (GMOs) are stably increasing and demonstrates LAMP is a potential and promising method for on spot identification of GMOs. However, little information is known for detection of GM potato events by LAMP. In this report, we developed an optimized and visual LAMP assay with high specificity and sensitivity to rapidly amplify genomic DNA of potato EH92-527-1 within 45 min. The limit of detection of LAMP in our study is 10-fold higher than the conventional PCR. Furthermore, LAMP products can be directly observed via naked eyes by addition of SYBR Green I without gel electrophoresis analysis and PCR-based equipment. Therefore, the LAMP assay developed in this paper provides an efficient, convenient and cost-effective tool for the detection of GM potato EH92-527-1.

    更新日期:2019-12-04
  • High-throughput sequencing reveals biofluid exosomal miRNAs associated with immunity in pigs
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : 2019-09-04
    Jie Zhang, Hui Luo, Zibiao Xiong, Kun Wan, Qinfeng Liao, Hang He

    Large numbers of miRNAs are found in biofluid exosomes. We isolated ~50–200 nm diameter exosomes from four types of porcine biofluid (urine, plasma, semen, and bile) using serial centrifugation and ultracentrifugation procedures. A total of 42.15 M raw data were generated from four small RNA libraries. This produced 40.17 M map-able sequences, of which we identified 204 conserved miRNAs, and 190 novel candidate miRNAs. Furthermore, we identified 34 miRNAs specifically expressed in only one library, all with well-characterized immune-related functions. A set of five universally abundant miRNAs (miR-148a-3p, miR-21-5p, let-7f-5p, let-7i-5p, and miR-99a-5p) across all four biofluids was also found. Function enrichment analysis revealed that the target genes of the five ubiquitous miRNAs are primarily involved in immune and RNA metabolic processes. In summary, our findings suggest that porcine biofluid exosomes contain a large number of miRNAs, many of which may be crucial regulators of the immune system.

    更新日期:2019-12-04
  • Berberine and palmatine inhibit the growth of human rhabdomyosarcoma cells
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : 2019-08-29
    Sayaka Shinji, Shunichi Nakamura, Yuma Nihashi, Koji Umezawa, Tomohide Takaya

    A natural isoquinoline alkaloid, berberine, has been known to exhibit anti-tumor activity in various cancer cells via inducing cell cycle arrest. However, it has not been investigated whether berberine and its analogs inhibit the growth of rhabdomyosarcoma (RMS), which is the most frequent soft tissue tumor in children. The present study examined the anti-tumor effects of berberine and palmatine on expansions of three human embryonal RMS cell lines; ERMS1, KYM1, and RD. Intracellular incorporation of berberine was relatively higher than that of palmatine in every RMS cell line. Berberine significantly inhibited the cell cycle of all RMS cells at G1 phase. On the other hand, palmatine only suppressed the growth of RD cells. Both of berberine and palmatine strongly inhibited the growth of tumorsphere of RD cells in three-dimensional culture. These results indicate that berberine derivatives have the potential of anti-tumor drugs for RMS therapy. Abbreviations: ARMS: alveolar rhabdomyosarcoma; ERMS: embryonal rhabdomyosarcoma; RMS: rhabdomyosarcoma

    更新日期:2019-12-04
  • CCA1 and LHY contribute to nonhost resistance to Pyricularia oryzae (syn. Magnaporthe oryzae) in Arabidopsis thaliana
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : 2019-09-03
    Saaya Yamaura, Yuri Yamauchi, Motoi Makihara, Takafumi Yamashino, Atsushi Ishikawa

    The circadian clock enables plants to adapt to their environment and control numerous physiological processes, including plant–pathogen interactions. However, it is unknown if the circadian clock controls nonhost resistance (NHR) in plants. To find out, we analyzed microarray data with the web-based tool DIURNAL to reveal that NHR-related genes show rhythmic expression patterns in the absence of a pathogen challenge. Our clock mutant analyses found that cca1-1 lhy-11 double mutant showed compromised NHR to Pyricularia oryzae, suggesting that two components of the circadian clock, CCA1 and LHY, are involved in regulating penetration resistance in Arabidopsis thaliana. By analyzing pen2 double mutants, we revealed that CCA1 contributes to time-of-day-dependent penetration resistance as a positive regulator and that LHY regulates post-penetration resistance as a positive regulator. Taken together, our results suggest that the circadian clock regulates the time-of-day-dependent NHR to P. oryzae and thus enables A. thaliana to counteract pathogen attacks. Abbreviations: EE: evening element; ETI: effector-triggered immunity; NHR: nonhost resistance; PAMP: pathogen-associated molecular pattern; PTI: PAMP-triggered immunity; SAR: systemic acquired resistance.

    更新日期:2019-12-04
  • The CEACAM1-derived peptide QLSN impairs collagen-induced human platelet activation through glycoprotein VI
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : 2019-09-05
    Yujia Ye, Wen Wan, Jing Wang, Wei Hu, Huawei Wang, Longjun Li, Peng Sang, Yajuan Gu, Deng Li, Zhe Wang, Zhaohui Meng

    Carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1) regulates collagen-mediated platelet activation through its cytoplasmic immunoreceptor tyrosine-based inhibition motifs (ITIMs). However, the function of CEACAM1’s extracellular cleavage fragments is currently unknown. In the present study, we used mass spectrometry (MS) to identify 9 cleavage fragments shed by matrix metallopeptidase 12 (MMP-12), and then we synthesized peptides with sequences corresponding to the fragments. QLSNGNRTLT (QLSN), a peptide from the A1-domain of CEACAM1, significantly attenuated collagen-induced platelet aggregation. QLSN also attenuated platelet static adhesion to collagen. Additionally, QLSN reduced human platelet secretion and integrin αIIbβ3 activation in response to glycoprotein VI (GPVI)–selective agonist, convulxin. Correspondingly, QLSN treatment significantly decreased convulxin-mediated phosphorylation of Src, protein kinase B (Akt), spleen tyrosine kinase (Syk) and phospholipase Cγ2 (PLCγ2) in human platelets. These data indicate that the CEACAM1-derived peptide QLSN inhibits GPVI-mediated human platelet activation. QLSN could potentially be developed as a novel antiplatelet agent.

    更新日期:2019-12-04
  • Aspartate racemase and D-aspartate in starfish; possible involvement in testicular maturation
    Biosci. Biotechol. Biochem. (IF 1.297) Pub Date : 2019-09-03
    Kimihiko Shibata, Noriko Sugaya, Yuko Kuboki, Hiroko Matsuda, Katsumasa Abe, Shouji Takahashi, Yoshio Kera

    D-Aspartate, aspartate racemase activity, and D-aspartate oxidase activity were detected in tissues from several types of starfish. Aspartate racemase activity in male testes of Patiria pectinifera was significantly elevated in the summer months of the breeding season compared with spring months. We also compared aspartate racemase activity with the gonad index and found that activity in individuals with a gonad index ≥6% was four-fold higher than that of individuals with a gonad index <6%. The ratio of the D-form of aspartate to total aspartate was approximately 25% in testes with a gonad index <6% and this increased to approximately 40% in testes with a gonad index ≥6%. However, such changes were not observed in female ovaries. Administration of D-aspartate into male starfish caused testicular growth. These results indicate the possible involvement of aspartate racemase and D-aspartate in testicular maturation in echinoderm starfish.

    更新日期:2019-12-04
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