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Foods ◽  
2021 ◽  
Vol 10 (12) ◽  
pp. 3064
Author(s):  
Qi Gao ◽  
Jia-Le Wu ◽  
Lan-Ping Jiang ◽  
Su-Qi Sun ◽  
Xue-Jun Gu ◽  
...  

Sweet potato plants were treated with selenium (Se). Spraying Se on the sweet potato leaves was an effective Se enrichment method and proteins were extracted from the sweet potato stem. The structural characteristics of the protein were investigated. Fourier transform infrared spectroscopy (FT-IR) detected more signals from the Se-enriched sweet potato stem protein (SSP), and the number of forms of Se chemical bonds gradually increased with increasing Se content, such as the Se-O bond in high Se-enriched SSP, indicating altered secondary structures.Scanning electron microscopy-energy dispersive spectrometry (SEM-EDS) indicated more Se atoms in the Se-enriched SSPs (SSSPs). The DSC results revealed that Se enrichment enhanced the thermal stability of the samples. Moreover, selenomethionine (SeMet), selenocystine (SeCys2), and methylselenocysteine (MeSeCys) were determined to be the main Se forms in the SSSPs. Furthermore, the SSSPs showed relatively higher superoxide anion radical and DPPH radical scavenging activities than the blank, which indicates that SSSPs can be used as antioxidants. By recovering the proteins, the agricultural by-product—sweet potato stem can be further utilized, and the obtained Se-enriched proteins may contribute to human health.


Blood ◽  
2021 ◽  
Vol 138 (Supplement 1) ◽  
pp. 2181-2181
Author(s):  
Hiroki Mizumaki ◽  
Dung Cao Tran ◽  
Kohei Hosokawa ◽  
Kazuyoshi Hosomichi ◽  
Hiroyuki Takamatsu ◽  
...  

Abstract [Background] Minor populations (0.003%-1.0%) of glycosylphosphatidylinositol-anchored protein-deficient granulocytes (GPI[-] Gs) are often detected in the peripheral blood (PB) of patients with acquired aplastic anemia (AA) and low-risk myelodysplastic syndromes and are thought to represent immune pathophysiology of bone marrow failure. We previously reported that minor GPI(-) G populations were detected in some healthy individuals (HIs) and persisted over several years at similar percentages (Katagiri T, et al. Stem Cells 2013). Several lines of evidence have suggested that small numbers of GPI(-) Gs detected in HIs are polyclonal populations mostly derived from short-lived PIGA-mutated committed progenitor cells. Minor GPI(-) G populations in AA patients may also be derived from multiple committed progenitor cells rather than from a few hematopoietic stem progenitor cells (HSPCs) with PIGA mutations. However, minor GPI(-) G populations usually persist for a long period of time at similar frequencies in AA patients, suggesting that they may instead be derived from a few HSPCs that have undergone PIGA mutations. This issue remains debated due to the inability to sequence the PIGA gene in the very few GPI(-) granulocytes available. We recently developed a sensitive method capable of detecting PIGA mutations in minor GPI(-) Gs using amplicon sequencing of GPI(-) Gs that were enriched with magnetic microbeads followed by FACS sorting. Using this method, we addressed whether minor GPI(-) G populations in AA patients and HIs are oligoclonal or polyclonal as well as which cell population they are derived from HSPCs or committed progenitor cells. [Methods] Five AA patients possessing 0.025%-0.898% GPI(-) Gs, 3 HIs who were found to have ≥0.003% (0.006%, 0.051%, and 0.059%) GPI(-) Gs during a screening of more than 200 HIs for GPI(-) Gs, 30 HIs (median: 37 years old, male/female:17/13) with 0% to 0.002% GPI(-) Gs, and 8 cord blood (CB) samples were subjected to enrichment of GPI(-) Gs for PIGA sequencing. Their leukocytes were treated with PE-labelled anti-CD55 monoclonal antibodies (mAbs) and anti-CD59 mAbs, and CD55 +CD59 + granulocytes were removed using magnetic microbeads labelled with anti-PE mAbs. CD11b +FLAER-negative granulocytes were sorted from the remaining granulocytes using FACSAria Fusion. DNA from sorted GPI(-) Gs was amplified using primers covering all exons of PIGA. Nucleotide sequences of the PIGA gene in GPI(-) Gs were determined using a next-generation sequencer. [Results] This novel enrichment method enabled the detection of only 1-4 different PIGA mutations in all 5 female AA patients (AA 1-5) with the total of different VAFs in each case reaching nearly 50% (Table 1). Limited kinds of PIGA mutations were also detected in three HIs (two males [HI 1 and 3] and one female [HI 2]). For HI 1 and HI 3, the VAFs of predominant PIGA-mutated sequences were longitudinally measurable using whole-blood DNA samples with droplet digital PCR, which showed no apparent changes in the VAF (0.020%-0.027% for HI 1 and 0.012%-0.025% for HI 3 over 4 and 6 years, respectively). The presence of mono or oligoclonal GPI(-) Gs in the 3 HIs prompted us to study 30 HIs who had been judged to be negative for minor GPI(-) G populations by a high-sensitivity flow cytometry method. The enrichment method unexpectedly identified clear CD11b highFLAER - GPI(-) G clusters in granulocytes from 24 of the 30 HIs (Figure 1a). The median number of GPI(-) Gs contained in 7 ml of PB was 31 (range, 1-136 cells). Sufficient amounts of DNA for NGS were obtained from sorted GPI(-) Gs of six subjects, and PIGA amplicon sequencing revealed 1-3 different PIGA mutations in four of the six subjects. The examination of fresh CB also revealed clear GPI(-) G clusters in four of eight samples (Figure 1b). PIGA amplicon sequencing of 79 GPI(-) Gs obtained from 1 male CB sample (CB 1) showed a sole PIGA mutation with VAFs of 95% (Figure 1c). [Conclusion] Minor GPI(-) G populations detectable in patients with AA and HIs are derived from a few PIGA-mutated HSPCs, not from committed myeloid progenitor cells, a finding that negates a hypothesis that a few PIGA-mutated HSPCs are selected from polyclonal PIGA-mutated HSPCs during transition from AA to florid PNH. Very small numbers of GPI(-) Gs are present much more frequently in HIs than previously thought and may also be derived from a few HSPCs with PIGA mutations that occur in HSPCs during the fetal stage. Figure 1 Figure 1. Disclosures Takamatsu: Bristol-Myers Squibb: Honoraria, Research Funding; Adaptive Biotechnologies, Eisai: Honoraria; SRL: Consultancy; Janssen: Consultancy, Honoraria, Research Funding. Yamazaki: Novartis Pharma: Honoraria; Kyowa Kirin: Research Funding; Kyowa Kirin: Honoraria. Nakao: Symbio: Consultancy; Kyowa Kirin: Honoraria; Novartis Pharma: Honoraria; Alexion Pharma: Research Funding.


2021 ◽  
Vol 2021 ◽  
pp. 1-11
Author(s):  
Li Hui ◽  
Yang Tao ◽  
Mao Xinmin

Objective. Diabetic retinopathy (DR) is one of the main diabetic microvascular complications in clinical practice, which features a complicated mechanism and insignificant efficacy. Therefore, it is urgent to find effective drugs. Xinjiang Coreopsis tinctoria Nutt is a rare alpine wild plant with unique effects and extremely high medicinal value. Preliminary studies have shown that it can reduce elevated blood sugar, unhealthy lipids, and antioxidants. This study was intended to investigate the protective effect of the effective components of Coreopsis tinctoria Nutt on the retinopathy of db/db diabetic mice and provide experimental basis for exploring the efficacy of Coreopsis tinctoria Nutt and the development of new drugs for the treatment of DR. Method. The db/db diabetic mouse models were used, and the effective components of Coreopsis tinctoria Nutt were obtained from Coreopsis tinctoria Nutt using macroporous resin enrichment method after alcohol extraction. These mice were divided into the normal group, model group, and high-dose Coreopsis tinctoria Nutt groups, the positive drug metformin group, and the metformin and Coreopsis tinctoria Nutt combination group. After these db/db type 2 diabetes mouse models were intervened for 10 weeks, their weight, blood sugar, glycosylated hemoglobin, serum MDA, SOD, and other indicators of each group were tested, and the expression changes of VEGF, ICAM1, PEDF, Bcl-2 in mouse retina were observed by immunohistochemistry method. Result. The effective components of Coreopsis tinctoria Nutt were obtained using macroporous resin enrichment method after alcohol extraction, which were mainly comprised of chlorogenic acid, flavone mariside, mariside, dicaffeoyl quinic acid, and flavone oxanine, with a total content of 532.82 mg/g, and the total flavonoid content of 330 mg/g. The effective components of Coreopsis tinctoria Nutt significantly reduced blood sugar and glycosylated hemoglobin and improved oxidative stress levels in db/db diabetic mice. Meanwhile, they reduced the expression of VEGF and ICAM1 in retinopathy and increased the expression of Bcl-2 and PEDF. The combination of Coreopsis tinctoria Nutt and metformin has the most significant effect. Conclusion. Coreopsis tinctoria Nutt can prevent and treat early diabetic retinopathy by affecting the expression of retinopathy-related factors.


Author(s):  
Seonjeong Lee ◽  
Shinyeong Ju ◽  
Seok Jin Kim ◽  
Jin-Oh Choi ◽  
Kihyun Kim ◽  
...  
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2021 ◽  
Vol 99 ◽  
pp. 103818
Author(s):  
Surendra Rasamsetti ◽  
Mark Berrang ◽  
Nelson A. Cox ◽  
Nikki W. Shariat

Author(s):  
Jie Chen ◽  
Zhigang Jiao ◽  
Jianwen Mo ◽  
Defa Huang ◽  
Zhengzhe Li ◽  
...  

AbstractA potential use of small extracellular vesicles (sEVs) for diagnostic and therapeutic purposes has recently generated a great interest. sEVs, when purified directly from various tissues with proper procedures, can reflect the physiological and pathological state of the organism. However, the quality of sEV is affected by many factors during isolation, including separation of sEV from cell and tissues debris, the use of enzymes for tissue digestion, and the storage state of tissues. In the present study, we established an assay for the isolation and purification of liver cancer tissues-derived sEVs (tdsEVs) and cultured explants-derived sEVs (cedsEVs) by comparing the quality of sEVs derived from different concentration of digestion enzyme and incubation time. The nano-flow cytometry (NanoFCM) showed that the isolated tdsEVs by our method are purer than those obtained from differential ultracentrifugation. Our study thus establishes a simple and effective approach for isolation of high-quality sEVs that can be used for analysis of their constituents. Graphical abstract


Insects ◽  
2021 ◽  
Vol 12 (9) ◽  
pp. 832
Author(s):  
Wei Lu ◽  
Xinhui Lan ◽  
Tong Zhang ◽  
Hao Sun ◽  
Sanyuan Ma ◽  
...  

To study the evolution of gene function and a species, it is essential to characterize the tandem repetitive sequences distributed across the genome. Cas9-based enrichment combined with nanopore sequencing is an important technique for targeting repetitive sequences. Cpf1 has low molecular weight, low off-target efficiency, and the same editing efficiency as Cas9. There are numerous studies on enrichment sequencing using Cas9 combined with nanopore, while there are only a few studies on the enrichment sequencing of long and highly repetitive genes using Cpf1. We developed Cpf1-based enrichment combined with ONT sequencing (CEO) to characterize the B. mori FibH gene, which is composed of many repeat units with a long and GC-rich sequence up to 17 kb and is not easily amplified by means of a polymerase chain reaction (PCR). CEO has four steps: the dephosphorylation of genomic DNA, the Cpf1 targeted cleavage of FibH, adapter ligation, and ONT sequencing. Using CEO, we determined the fine structure of B. moriFibH, which is 16,845 bp long and includes 12 repetitive domains separated by amorphous regions. Except for the difference of three bases in the intron from the reference gene, the other sequences are identical. Surprisingly, many methylated CG sites were found and distributed unevenly on the FibH repeat unit. The CEO we established is an available means to depict highly repetitive genes, but also a supplement to the enrichment method based on Cas9.


2021 ◽  
Vol 9 ◽  
Author(s):  
Yun Cui ◽  
Xuefang Dong ◽  
Xiaofei Zhang ◽  
Cheng Chen ◽  
Dongmei Fu ◽  
...  

HKU1 is a human beta coronavirus and infects host cells via highly glycosylated spike protein (S). The N-glycosylation of HKU1 S has been reported. However, little is known about its O-glycosylation, which hinders the in-depth understanding of its biological functions. Herein, a comprehensive study of O-glycosylation of HKU1 S was carried out based on dual-functional histidine-bonded silica (HBS) materials. The enrichment method for O-glycopeptides with HBS was developed and validated using standard proteins. The application of the developed method to the HKU1 S1 subunit resulted in 46 novel O-glycosylation sites, among which 55.6% were predicted to be exposed on the outer protein surface. Moreover, the O-linked glycans and their abundance on each HKU1 S1 site were analyzed. The obtained O-glycosylation dataset will provide valuable insights into the structure of HKU1 S.


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