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- Cheng, Yirui, et al.
(författare)
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Comparison of serum exosome isolation methods on co-precipitated free microRNAs.
- 2020
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Ingår i: PeerJ. - : PeerJ. - 2167-8359. ; 8
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Tidskriftsartikel (refereegranskat)abstract
- Background: Exosomes are nano-sized extracellular vesicles containing different biomolecules such as proteins and microRNAs (miRNAs) that mediate intercellular communication. Recently, numerous studies have reported the important functions of exosomal miRNAs in disease development and the potential clinical application as diagnostic biomarkers. Up to now, the most commonly used methods to extract exosomes are ultracentrifugation (UC) and precipitation-based commercial kit (e.g., ExoQuick). Generally, both UC and ExoQuick method could co-isolate contaminating proteins along with exosomes, with the UC method yielding even purer exosomes than ExoQuick. However, the comparison of these two methods on co-precipitated free miRNAs is still unknown.Methods: In this study, we isolated exosomes from the human serum with exogenously added cel-miR-39 by UC and ExoQuick and compared the proportion of cel-miR-39 co-precipitated with exosomes extracted by these two methods.Results: Using exogenous cel-miR-39 as free miRNAs in serum, we concluded that ExoQuick co-isolates a small proportion of free miRNAs while UC hardly precipitates any free miRNAs. We also found that incubation at 37 °C for 1 h could decrease the proportion of free miRNAs, and exosomal miRNAs like miR-126 and miR-152 also decreased when RNase A was used. In conclusion, our findings provide essential information about the details of serum exosome isolation methods for further research on exosomal miRNAs.
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2. |
- Cheng, Yirui, et al.
(författare)
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NDFIP1 limits cellular TAZ accumulation via exosomal sorting to inhibit NSCLC proliferation
- 2023
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Ingår i: Protein & Cell. - : Springer Nature. - 1674-800X .- 1674-8018. ; 14:2, s. 123-136
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Tidskriftsartikel (refereegranskat)abstract
- NDFIP1 has been previously reported as a tumor suppressor in multiple solid tumors, but the function of NDFIP1 in NSCLC and the underlying mechanism are still unknown. Besides, the WW domain containing proteins can be recognized by NDFIP1, resulted in the loading of the target proteins into exosomes. However, whether WW domain-containing transcription regulator 1 (WWTR1, also known as TAZ) can be packaged into exosomes by NDFIP1 and if so, whether the release of this oncogenic protein via exosomes has an effect on tumor development has not been investigated to any extent. Here, we first found that NDFIP1 was low expressed in NSCLC samples and cell lines, which is associated with shorter OS. Then, we confirmed the interaction between TAZ and NDFIP1, and the existence of TAZ in exosomes, which requires NDFIP1. Critically, knockout of NDFIP1 led to TAZ accumulation with no change in its mRNA level and degradation rate. And the cellular TAZ level could be altered by exosome secretion. Furthermore, NDFIP1 inhibited proliferation in vitro and in vivo, and silencing TAZ eliminated the increase of proliferation caused by NDFIP1 knockout. Moreover, TAZ was negatively correlated with NDFIP1 in subcutaneous xenograft model and clinical samples, and the serum exosomal TAZ level was lower in NSCLC patients. In summary, our data uncover a new tumor suppressor, NDFIP1 in NSCLC, and a new exosome-related regulatory mechanism of TAZ.
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3. |
- Joshi, Prachi, et al.
(författare)
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Approaches for Assigning Offsets to Signals for Improving Frame Packing in CAN-FD
- 2020
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Ingår i: IEEE Transactions on Computer-Aided Design of Integrated Circuits and Systems. - : Institute of Electrical and Electronics Engineers (IEEE). - 0278-0070 .- 1937-4151. ; 39, s. 1109-1122
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Tidskriftsartikel (refereegranskat)abstract
- Controller area network (CAN) is a widely used protocol that allows communication among electronic control units (ECUs) in automotive electronics. It was extended to CAN with flexible data-rate (CAN-FD) to meet the increasing demand for bandwidth generated by the growing number of features in modern automobiles. The signal-to-frame packing problem has been studied in the literature for both CAN and CAN-FD. In this paper, we propose and formulate the signal offset assignment problem (SOAP) in CAN-FD to improve the bus utilization during frame packing. We propose two algorithmic themes to solve SOAP and establish their worst case performance guarantees. The first is a general approximation framework (GAF) which can use any approximation algorithm for the makespan minimization problem (MMP) in multiprocessor systems. Its performance guarantee is the product of the performance guarantee of the MMP algorithm and the number of distinct periods in the frame. The second is a 2-D strip packing-based framework (2DSPF) which uses the bottom left fill algorithm for 2-D strip packing. The performance guarantee is 2G , where G is the minimum number of groups into which the set of signals can be partitioned so that the periods of the signals in the same group form a geometric series. The experimental results for GAF and 2DSPF indicate that by carefully assigning offsets for signals in frame packing schemes, one can achieve about 10.83% improvement in bus utilization in CAN-FD systems.
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