マウス原腸胚形成におけるCables2の役割

筑波大学 (University of Tsukuba)201

Enzimatska obrada iskorištenih listića zelenog čaja i njihova primjena u proizvodnji čajnog peciva s velikim udjelom prehrambenih vlakana

Research background. By-products of food industry have been studied as sources of high fibre and antioxidant ingredients for healthy food products, because of their economic and environmental benefits. However, the soluble dietary fibre content of these materials is usually lower than the recommended value that is claimed to bring positive health effects. Enzymatic treatment could be an efficient method for modifying insoluble and soluble dietary fibre contents of these materials. The purpose of this study is to investigate the effects of enzymatic treatment conditions on soluble, insoluble and total dietary fibre mass fractions in spent green tea leaves, and evaluate the quality of dough and cookies when different mass fractions of untreated and treated leaves were added to the recipe. Experimental approach. The mass fractions of soluble, insoluble and total dietary fibre in spent tea leaf powder was evaluated after the leaves were treated with cellulase amount of 0−25 U/g for 0 to 2 h. Wheat flour was replaced by untreated and treated spent tea leaf powder at 0, 10, 20, 30 and 40% in cookie formulation. Textural properties of dough, proximate composition, physical properties and overall acceptability of cookies were analysed. Results and conclusions. The appropriate conditions for enzymatic treatment were enzyme loading of 20 U/g and biocatalytic time of 1.5 h, under which the mass fraction of soluble dietary fibre in spent tea leaves increased by 144.5% compared to that of the control sample. The addition of spent tea leaves led to the increase in dough hardness. Increase in the spent tea leaf amount also enhanced fibre mass fraction, antioxidant activity and hardness of cookies but reduced their overall acceptability. Moreover, the enzymatic treatment of spent tea leaves improved the soluble to total dietary fibre ratio of the cookies, which influenced their textural properties and health benefits. The cookies with added 20% untreated or treated spent tea leaves were overall accepted by the panel. Novelty and scientific contribution. For the first time, spent tea leaves have been treated with enzymes to improve their soluble to total dietary fibre ratio. The treated spent tea leaves are a new promising high-fibre antioxidant ingredient for cookie preparation.Pozadina istraživanja. Nusproizvodi prehrambene industrije ispituju se iz ekonomskih i ekoloških razloga kao izvori sastojaka s velikim udjelom prehrambenih vlakana i antioksidacijskim učinkom koji se mogu upotrijebiti u proizvodnji zdrave hrane. Međutim, udjel topljivih prehrambenih vlakana u nusproizvodima je obično manji od preporučenih vrijednosti koje mogu imati pozitivan učinak na zdravlje. Enzimatska obrada bi mogla biti učinkovita metoda modificiranja netopljivih i topljivih prehrambenih vlakana iz otpada prehrambene industrije. Svrha je ovog rada bila ispitati utjecaj različitih uvjeta enzimatske obrade na masene udjele topljivih, netopljivih i ukupnih prehrambenih vlakana u iskorištenim listićima zelenog čaja, te procijeniti kakvoću tijesta i čajnog peciva s dodatkom različitih masenih udjela obrađenih i neobrađenih listića. Eksperimentalni pristup. Maseni udjeli topljivih, netopljivih i ukupnih prehrambenih vlakana u prahu od iskorištenih listića zelenog čaja mjereni su nakon njihove obrade s 0–25 U/g celulaze tijekom 0 do 2 h. U smjesi za čajno pecivo je pšenično brašno zamijenjeno s 0, 10, 20, 30 i 40 % obrađenog ili neobrađenog praha iskorištenih listića zelenog čaja. Ispitani su tekstura tijesta te kemijski sastav, fizikalna svojstva i ukupna prihvatljivost dobivenog čajnog peciva. Rezultati i zaključci. Pri povoljnim uvjetima enzimatske obrade, a to su 20 U/g enzima i vrijeme trajanja biokatalitičkog procesa od 1,5 sata, maseni udjel topljivih prehrambenih vlakana u iskorištenim listićima zelenog čaja porastao je za 144,5 % u usporedbi s kontrolnim uzorkom. Dodatkom iskorištenih listića zelenog čaja povećala se tvrdoća tijesta. Povećanjem količine listića čaja povećali su se i maseni udjel vlakana, antioksidacijska svojstva i tvrdoća čajnog peciva, ali se smanjila njihova prihvatljivost. Osim toga, enzimatskom se obradom iskorištenih listića zelenog čaja poboljšao omjer topljivih i ukupnih vlakana u čajnom pecivu, što je utjecalo na njegovu teksturu i pozitivan učinak na zdravlje. Panel ocjenjivača je okarakterizirao čajno pecivo s dodatkom 20 % obrađenih ili neobrađenih iskorištenih listića čaja kao prihvatljive. Novina i znanstveni doprinos. Prvi put je enzimatskom obradom iskorištenih listića zelenog čaja poboljšan njihov omjer topljivih i ukupnih prehrambenih vlakana. Obrađeni iskorišteni listići zelenog čaja novi su obećavajući sastojak s antioksidacijskim svojstvima koji se može upotrijebiti u pripremi čajnog peciva

HMU fluorinze mouthwash enhances enamel remineralization: An in vitro study

BACKGROUND: Fluoride therapy has long been used extensively to prevent dental caries. Fluoride appears in variety of dental care products such as mouthrinse, dentifrice, gel, etc. HMU fluorinze is the first mouthwash containing fluoride in Vietnam. AIM: This research was conducted to evaluate the efficacy of HMU Fluorinze mouthwash on remineralizing enamel in laboratory conditions. METHODS: 20 third molars teeth were cleaned and covered with nail polish , except for a 3x3 mm square on their buccal surfaces. These teeth went through two steps: demineralization using Coke and remineralization for 20 days: 1) using standard calcifying solution (control group) and 2) using standard calcifying solution + HMU Fluorinze mouthwash 2 times/day (experimental group). The mineralization index of enamel structure after demineralization and remineralization was assessed by DIAGNOdent pen 2190. RESULTS: The mineralization indexes of the control group and experimental group at baseline were 3.65 ± 0.76 and 3.35 ± 0.64, after demineralization were in turn of 21.78 ± 4.48 and 20.25 ± 2.26; and after remineralization were 6.30 ± 1.03 and 3.90 ± 1.24. The different figures  between the two groups after remineralization shows statistical significance (p<0.01). Group B using HMU fluorinze mouthwash after 20 days did not differ from the original results (p = 0.272), in contrast with the control group (p<0.01). CONCLUSIONS: HMU fluorinze mouthwash has better mineralization effect than standard calcifying solution

Differential Cytotoxicity of Curcumin-Loaded Micelles on Human Tumor and Stromal Cells

peer reviewedAlthough curcumin in the form of nanoparticles has been demonstrated as a potential anti-tumor compound, the impact of curcumin and nanocurcumin in vitro on normal cells and in vivo in animal models is largely unknown. This study evaluated the toxicity of curcumin-loaded micelles in vitro and in vivo on several tumor cell lines, primary stromal cells, and zebrafish embryos. Breast tumor cell line (MCF7) and stromal cells (human umbilical cord vein endothelial cells, human fibroblasts, and human umbilical cord-derived mesenchymal stem cells) were used in this study. A zebrafish embryotoxicity (FET) assay was conducted following the Organisation for Economic Co-operation and Development (OECD) Test 236. Compared to free curcumin, curcumin PM showed higher cytotoxicity to MCF7 cells in both monolayer culture and multicellular tumor spheroids. The curcumin-loaded micelles efficiently penetrated the MCF7 spheroids and induced apoptosis. The nanocurcumin reduced the viability and disturbed the function of stromal cells by suppressing cell migration and tube formation. The micelles demonstrated toxicity to the development of zebrafish embryos. Curcumin-loaded micelles demonstrated toxicity to both tumor and normal primary stromal cells and zebrafish embryos, indicating that the use of nanocurcumin in cancer treatment should be carefully investigated and controlled.3. Good health and well-bein

Disruption of entire Cables2 locus leads to embryonic lethality by diminished Rps21 gene expression and enhanced p53 pathway

In vivo function of CDK5 and Abl enzyme substrate 2 (Cables2), belonging to the Cables protein family, is unknown. Here, we found that targeted disruption of the entire Cables2 locus (Cables2d) caused growth retardation and enhanced apoptosis at the gastrulation stage and then induced embryonic lethality in mice. Comparative transcriptome analysis revealed disruption of Cables2, 50% down-regulation of Rps21 abutting on the Cables2 locus, and up-regulation of p53-target genes in Cables2d gastrulas. We further revealed the lethality phenotype in Rps21-deleted mice and unexpectedly, the exon 1-deleted Cables2 mice survived. Interestingly, chimeric mice derived from Cables2d ESCs carrying exogenous Cables2 and tetraploid wild-type embryo overcame gastrulation. These results suggest that the diminished expression of Rps21 and the completed lack of Cables2 expression are intricately involved in the embryonic lethality via the p53 pathway. This study sheds light on the importance of Cables2 locus in mouse embryonic development.Grant-in-Aid for Scientific Research(B), Japan Society for the Promotion of Science (JSPS KAKENHI 17H03568) Fumihiro Sugiyama Grant-in-Aid for Scientific Research(S), Japan Society for the Promotion of Science (JSPS KAKENHI 26221004) Satoru Takahashi Grant-in-Aid for Scientific Research(C), Japan Society for the Promotion of Science (JSPS KAKENHI 17K07130) Hiroyoshi Iseki Grant-in-Aid for Young Scientists (B), Japan Society for the Promotion of Science (JSPS KAKENHI 19K16020) Tra Thi Huong Dinh Grant-in-Aid for Scientific Research(A), Japan Society for the Promotion of Science (JSPS KAKENHI 20H00444) Fumihiro Sugiyama The Cooperative Research Project Program of Life Science Center for Survival Dynamics, Tsukuba Advanced Research Alliance (TARA Center), University of Tsukuba, Japan (182107) Fumihiro Sugiyam

ĐA HÌNH GEN LEPTIN VÀ THYROGLOBULIN LIÊN QUAN ĐẾN TÍNH TRẠNG NĂNG SUẤT VÀ CHẤT LƯỢNG THỊT Ở TỔ HỢP BÒ LAI SENEPOL × LAI BRAHMAN NUÔI TẠI TỈNH THỪA THIÊN HUẾ

This study was conducted to evaluate the polymorphisms in the intron 2 of leptin (LEP) gene (g.1926C>T) and in the 5’UTR of thyroglobulin (TG) gene (g.422C>T) in Senepol × lai Brahman crossbred cows raised in Thua Thien Hue province. The results of evaluating LEP and TG gene polymorphisms on 30 DNA samples with the PCR-RFLP technique show that the LEP/Sau3AI gene is polymorphic in the Senepol × lai Brahman crossbred cows, with the A allele related to high production and reproduction traits as the dominant allele (0.92). Five over 30 cows carried allele B related to high-fat yield percentage, and all were raised in Dien Mon commune. In contrast, all 30 studied cows carried heterozygous TG/PsuI genotype (CT). More DNA samples should be analysed to confirm the polymorphism of the TG5/PsuI gene in this crossbred cattle in Thua Thien Hue province, and further selection is needed to increase the frequency of the T allele related to a high marbling score of beef. These results are informative for further breeding of Senepol × lai Brahman crossbred cows.Nghiên cứu này được thực hiện nhằm đánh giá đa hình đoạn intron 2 của gen Leptin (LEP) (g.1926C>T) và vùng 5’UTR của gen thyroglobulin (TG) (g.422C>T) ở tổ hợp bò lai Senepol × lai Brahman nuôi tại tỉnh Thừa Thiên Huế. Kết quả đánh giá đa hình gen LEP/Sau3AI và TG5/PsuI trên 30 mẫu DNA bằng kỹ thuật PCR-RFLP cho thấy có sự đa hình về kiểu gen LEP/Sau3AI; tần số alen A liên quan đến các tính trạng năng suất và sức sinh sản cao là chủ yếu (0,92). Năm trong số 30 con bò mang alen B liên quan đến tỷ lệ mỡ trong thân thịt cao được phát hiện ở xã Điền Môn. Ngược lại, cả 30 con bò khảo sát đều mang kiểu gen TG5/PsuI dị hợp tử (CT). Do đó, để xác định chính xác hơn tính đa hình của gen TG5/PsuI trên tổ hợp bò lai Senepol × lai Brahman, cần tăng số lượng mẫu khảo sát và tiếp tục chọn lọc để tăng tần số alen T liên quan đến chỉ số vân mỡ cao trong thịt bò. Các kết quả nghiên cứu này cung cấp những thông tin di truyền ở cấp độ phân tử đầu tiên trên tổ hợp bò lai Senepol × lai Brahman nuôi tại tỉnh Thừa Thiên Huế và là cơ sở cho những nghiên cứu chọn lọc tiếp theo nhằm nâng cao chất lượng bò thịt

Generation of bicistronic reporter knockin mice for visualizing germ layers

Knockout mouse models are commonly used in developmental biology to investigate the functions of specific genes, and the knowledge obtained in such models has yielded insights into the molecular mechanisms underlying developmental processes. Gastrulation is the most dynamic process in embryogenesis during which differentiation into three germ layers occurs. However, the functions of genes involved in gastrulation are not completely understood. One major reason for this is the technical difficulty of embryo analysis to understand germ layer location. We have generated three reporter mouse strains in which the germ layers are distinguished by different fluorescent reporters. Using CRISPR/Cas9 genome editing in mouse zygotes, the fluorescent reporter genes, EGFP, tdTomato, and TagBFP including 2A peptide sequences were knocked into the appropriate sites before the stop codon of the Sox17 (endoderm marker), Otx2 (ectoderm marker), and T (mesoderm marker) genes, respectively. Founder mice were successfully generated in the Sox17-2A-EGFP, Otx2-2A-tdTomato, and T-2A-TagBFP knockin reporter strains. Further, homozygous knockin mice of all strains appeared morphologically normal and were fertile. On stereomicroscopic analysis, fluorescent signals were detected in a germ layer-specific manner from heterozygous embryos at embryonic day (E) 6.5–8.5 in all strains, and were immunohistochemically demonstrated to match their respective germ layer-specific marker protein at E7.5. Taken together, these observations suggest that the Sox17-2A-EGFP, Otx2-2A-tdTomato, and T-2A-TagBFP knockin reporter mice may be useful for comprehensive analysis of gene function in germ layer formation