Abstract In this study, the bioaccessibility and antioxidant activity of phenolic compounds in insoluble dietary fiber (IDF) and soluble dietary fiber (SDF) derived from hulless barley were evaluated by an in vitro gastrointestinal (GI) digestion model. The total phenolic and flavonoid contents, as well as antioxidant activity of phenolic compounds in IDF and SDF following GI digestion were studied. The results obtained showed an increase in total phenolic and flavonoid contents, as well antioxidant activity compared with undigested extracts. Moreover, the bioaccessibility indexes of phenolic compounds in IDF and SDF were 490.90 ± 3.10% and 1608.79 ± 40.63% respectively, after GI digestion. Similarly, the bioaccessibility indexes of flavonoids in IDF and SDF were 179.20 ± 15.16% and 814.36 ± 26.31%, respectively. Based on our findings, individual phenolic compounds show different stability in the digestion process. The content of ferulic acid has different trends in IDF and SDF during GI digestion. This study could provide a scientific basis for hulless barley DF as valuable food additives. Practical Application Hulless barley is a unique cereal with potential health benefits due to high dietary fiber (DF) content and phenolic compounds. Phenolic compounds could be linked to DF through chemical bonds. Phenolic compounds in DF can be slowly and continuously released under acidic, alkaline, and enzymatic conditions by in vitro gastrointestinal digestion, which could maintain a higher phenolic concentration in the bloodstream and be beneficial for human health. This study could provide a scientific basis for hulless barley DF as valuable food additives.
BACKGROUND: Despite the potential utility of primate somatic cell nuclear transfer (SCNT) to biomedical research and to the production of autologous embryonic stem (ES) cells for cell- or tissue-based therapy, a reliable method for SCNT is not yet available. Employing the rhesus monkey as a clinically relevant animal model, we have compared a conventional electrofusion method for SCNT with a one-step micromanipulation (OSM) method. METHODS: A prospective, randomized trial was conducted using only oocytes that were mature [metaphase II (MII)] at collection and a fibroblast-like cell line as nuclear donor cells (fetal fibroblasts). The embryos produced were characterized for in vitro developmental potential, cell number, karyotype and expression of nuclear mitotic apparatus (NuMA) and OCT-4. RESULTS: An in vitro blastocyst development rate of 24.4% was achieved with the OSM method, significantly higher than the 12.2% obtained following electrofusion. SCNT-produced embryos expressed normal karyotypes, cell numbers and NuMA and OCT-4 proteins in most cases. SCNT with male nuclear donor cells resulted in the production of male, SCNT blastocysts, eliminating the possibility of a parthenogenetic origin. Of the four fibroblast cell lines tested as nuclear donor cells, two supported the routine production of blastocysts following SCNT. CONCLUSIONS: The application of a modified SCNT technique (OSM) followed by embryo culture in hamster embryo culture medium-10 (HECM-10) allows, for the first time, the routine production of SCNT blastocysts, most of which appear normal by immunochemical, cytochemical and in vitro developmental criteria. These embryos will provide a resource for isolating ES cells and for studies of nuclear reprogramming by monkey cytoplasts.
Gene editing in non-human primates may lead to valuable models for exploring the etiologies and therapeutic strategies of genetically based neurological disorders in humans. However, a monkey model of neurological disorders that closely mimics pathological and behavioral deficits in humans has not yet been successfully generated. Microcephalin 1 (MCPH1) is implicated in the evolution of the human brain, and MCPH1 mutation causes microcephaly accompanied by mental retardation. Here we generated a cynomolgus monkey (Macaca fascicularis) carrying biallelic MCPH1 mutations using transcription activator-like effector nucleases. The monkey recapitulated most of the important clinical features observed in patients, including marked reductions in head circumference, premature chromosome condensation (PCC), hypoplasia of the corpus callosum and upper limb spasticity. Moreover, overexpression of MCPH1 in mutated dermal fibroblasts rescued the PCC syndrome. This monkey model may help us elucidate the role of MCPH1 in the pathogenesis of human microcephaly and better understand the function of this protein in the evolution of primate brain size.
Mistletoes are used medicinally in order to treat various human illnesses. Few studies have reported on the phenolic content and antioxidant properties of Chinese mistletoes (CMs). In this work, the total phenolic content (TPC), total flavonoid content (TFC), and antioxidant activities of soluble and insoluble-bound phenolic extracts from CMs hosted by Camellia assamica (Mast.) Chang (CMC) and Pyrus, i, f. (CMP) were compared. Phenolic compounds in CMC and CMP were identified and quantified using high-performance liquid chromatography (HPLC). The results indicated that the TPC of soluble phenolic extracts was higher than insoluble-bound phenolic counterparts in both CMC and CMP. In addition, the TPC of soluble, insoluble-bound and total phenolic fractions (9.91 ± 0.23, 4.59 ± 0.27 and 14.50 ± 0.35 μmol ferulic acid equivalents per gram (FAE/g) dry sample) extracted from CMP were higher than those extracted from CMC. The soluble phenolic extracts in CMP showed higher antioxidant activities than those in CMC. Eighteen phenolic compounds from soluble and insoluble-bound phenolic extracts from the CMs were identified and quantified by HPLC. This study indicates that CMC and CMP, especially the latter, could be sources of antioxidants in human health care.
The cryoprotective effect and toxicity of Bovine Serum Albumin (BSA) at 8 concentrations (0, 2, 4, 8, 16, 32, 64, 96 mg/mL, respectively) supplemented in cryoprotectant solutions on mouse pronuclear-stage embryo vitrification were studied. Survival rate, cleavage rate, blastocyst rate and mean cell numbers of blastocyst of vitrified embryos were used as criterion to evaluate the effect of BSA on the post-thawed embryos development. The results showed that no statistical differences of survival rate, cleavage rate, blastocyst rate and mean cell numbers of blastocyst of vitrified groups both in the toxic effect assay (P>0.05) and the vitrification assay (P>0.05). The results indicated that BSA in this cryoprotectant solutions has no effect on the viability of pronuclearstage embryos. Economical, practical and biosecure considerations do not support the use of BSA in vitrificationsolutions.
HBB-deficient Macaca fascicularis monkey presents with human β-thalassemiaDear Editor, β-Thalassemia is a common severe genetic disease caused by mutations in HBB and affects approximately 1.5% of the global population (Origa, 2017).In southern China, the carrier rate of β-thalassemia is as high as 6.43%, creating a high socio-economic burden (Xiong et al., 2010).In adult humans, there are three types of hemoglobin: HbA1 (∼97%), HbA2 (∼2%) and HbF (∼1%).HbA1 (α 2 β 2 ) is composed of two α-globin and two β-globin subunits encoded by HBA and HBB, respectively; HbF (α 2 γ 2 ) is made up of two α-globin subunits and two γ-globin subunits encoded by HBG.Mutations in the coding region or regulatory region of HBB are involved in β-thalassemia pathogenesis.Except for some rare dominant mutations, most HBB mutations are recessive (Origa, 2017).Depending on the mutation type, the β-globin level will either be reduced or completely depleted, resulting in α-globin accumulation and precipitation.These α-globin precipitates lead to red blood cell death, resulting in anemia and tissue damage, and even death in βthalassemia major patients.Blood transfusions can help slow disease progression but lead to iron overload, ultimately resulting in iron toxicity.Bone marrow transfer is the only cure in the clinic and is available only to a small percentage of patients with human leukocyte antigen-matched donors.Recently, gene therapy and gene editing therapy have shown great promise in curing β-thalassemia (Glaser et al., 2015;Thompson et al., 2018).However, no appropriate animal models are available for evaluating the safety and efficacy of such advanced therapeutic strategies in vivo.β-thalassemia mice are the sole animal model available for research.However, substantial differences have been reported between the types and expression patterns of human and mouse globins (McColl and Vadolas, 2016).Moreover, mice contain no fetal globin gene equivalent, and homozygous mutations of HBB in mouse for early models of β-thalassemia major or Cooley anemia are all embryonic lethal (Huo et al., 2009).Recently, significant phenotype and physiology differences have been reported between SIRT6null mice and the non-human primate model (Zhang et al., 2018).Thus, an appropriate non-human primate model is needed for human β-thalassemia studies and treatments.
Abstract Background: The restriction factor, TRIM5α is known to limit retroviral infection at an early post-entry stage in a species-specific manner. The current study was designed to assess the efficacy of TRIM5α in restricting the human immunodeficiency virus (HIV) using a pseudo-viral infection model in cynomolgus monkey embryonic fibroblast cells (CMEFs down the TRIM5α expression using the lentivirus short hairpin RNA (shRNA) knockdown vector system. Methods: Three shRNAs against the TRIM5α gene were designed and cloned into lentiviral vectors expressing the green fluorescent protein (GFP) and red fluorescent protein (RFP) reporter genes. The stable knockdown of TRIM5α gene expression was confirmed by both real-time RT-PCR as well as Western blot. The lentiviral inducible shRNA vector was then transduced into the CMEFs infected with HIV Pseudo-virus. Results: We successfully designed and cloned three shRNAs targets into lentiviral vectors to knock down the TRIM5α gene expression. Further, transduction of the shRNAs into the pseudo-viral HIV infected CMEFs showed greater susceptibility to the infection. Conclusion: Here, for the first time, we successfully knocked down the TRIM5α gene using RNA interference (RNAi) technology and found TRIM5α-null CMEF cells to be susceptible to HIV pseudo-virus infection. Our research provides an important strategy for the establishment of shRNA based lentiviral vector system for gene knockdown in the cynomolgus monkey animal model against HIV infection.
'/%3e%3cuse xlink:href='%23a' transform='rotate(23.036 .093 25.536)'/%3e%3cuse xlink:href='%23a' transform='rotate(45.87 1.273 16.18)'/%3e%3cuse xlink:href='%23a' transform='rotate(69.945 .996 12.078)'/%3e%3cuse xlink:href='%23a' transform='rotate(20.66 -19.689 31.932)'/%3e%3c/svg%3e)
