分类: 生物学 >> 生物物理学 >> 生物物理、生物化学与分子生物学 提交时间: 2016-05-05
摘要: Scintillation proximity assays (SPA), developed from the original radioimmunoassay, is a homogeneous, sensitive, fast and simple scintillation bead-based assay platform. Due to the diversification of radio-labeled ligands and of affinity tags modified on the scintillation carrier's surface, as well as the development of scintillation recorders and liquid handling technologies, SPA has been widely used as a high throughput method for lead compound screening and biomarker detection. In this review, after a brief introduction of the principle and operation of SPA, a detailed advance of its applications in life sciences were outlined, especially on membrane proteins analysis and at the cell level analysis with detailed classical reports. Beyond that, the methods on increasing the ratios of signal/noise were also analyzed. The wide spread of SPA, especially of its innovations at the cell level research will definitely promote our comprehensive understanding of cellular system biology.
分类: 地球科学 >> 地理学 提交时间: 2021-02-10 合作期刊: 《干旱区科学》
摘要: Crop residue is a major source of soil organic matter; therefore, application of crop straw to soil contributes to the sustainable development of organic agriculture. To better understand the transformation of crop straw in orchard soils, we investigated the relationship between the characteristics of straw decomposition and functional diversity of associated microbial communities in a long-term peach orchard, China. Mesh bags, each containing 30 g of corn or bean straw, were buried at a soil depth of 20 cm in a 12-year-old peach orchard for 360 d (October 2011–October 2012). Three treatments were applied, i.e., fresh corn straw, fresh corn straw with nitrogen fertilizer (urea, 10.34 g/kg), and fresh bean straw. Changes in straw residual rate, straw water content and soil conditions were monitored after treatment. The functional diversity of straw-associated microbial communities was analyzed by the Biolog-Eco microplate assay. During the decomposition process, straw residual rates did not vary considerably from 10 d (30.4%–45.4%) to 360 d (19.0%–30.3%). Irrespective of nitrogen addition, corn straw decomposed faster than bean straw. Corn straw with nitrogen fertilizer yielded the highest average well color development (AWCD) values (1.11–1.67), followed by corn straw (1.14–1.68) and bean straw (1.18–1.62). Although the AWCD values did not differ significantly among the three treatments, substantial differences occurred across various time periods of the decomposition process (P<0.01). In terms of carbon source utilization, the dominant microbial groups fed mainly on saccharides. Hard-to-decompose substances gradually accumulated in the middle and late stages of straw decomposition. Of the six categories of carbon sources tested, the utilization rate of aromatics was the lowest with corn straw, whereas that of polymers was the lowest with bean straw. Among different treatments, straw residual rate was negatively correlated to soil available phosphorous, soil available potassium and soil temperature (P<0.05), but not to soil water content. In some cases (corn straw with or without nitrogen fertilizer), straw residual rate was negatively correlated to straw water content, amino acid utilization and carboxylic acid utilization, and positively correlated with microbial species richness and evenness (P<0.05). Microbial community associated with corn and bean straw decomposition in soil was respectively dominated by aromatic- and polymer-metabolizing groups during the middle and late stages of this process, which could reduce the stability of microbial community structure and decrease the rate of straw decomposition in the fruit tree orchard.
分类: 核科学技术 >> 核材料与工艺技术 提交时间: 2023-06-18 合作期刊: 《Nuclear Science and Techniques》
摘要: We evaluated the protective effect of N-acetylcysteine (NAC) on immunity system irradiated by 12C6+ ion beam. Kun-Ming mice were whole-body irradiated by 12C6+ ion at doses of 0, 0.5, 1, 1.5, 2, 2.5 and 3 Gy. The results showed that in saline group, the lymphocytes DNA double-strand breaks (DSBs), maleic dialdehyde, thymocytes number in G0/G1 and apoptosis percentage increased with dose increment, and the levels of interferon-, glutathione, superoxide radical (SOD) and natural killer cells activity decreased with dose increment. However, there were no significant changes in NAC-treated group. The data indicated that pre-treatment with NAC could significantly remove the ROS by counteracting the glutamate, decrease excessive lipid peroxidation reaction and SOD damages, and protect DNA, lymphocytes and cytokines against irradiation.
分类: 核科学技术 >> 核材料与工艺技术 提交时间: 2023-06-18 合作期刊: 《Nuclear Science and Techniques》
摘要: This study investigated the impact of simulated microgravity on acute injury induced by low doses of carbon ions in male reproductive organs of mice, and determined alterations in spermatogenic function and expression levels of apoptotic factors in mice following exposure to acute irradiation after 7 days of simulated microgravity. The results demonstrated that significant reductions in spermatozoa, primary spermatocytes and spermatogonia, and increased globular cells in seminiferous tubule and pro-apoptotic proteins were observed in the group exposed to over 0.4 Gy irradiation. Collectively, the data suggest that lesions inflicted by simulated microgravity are not markedly modified by lower doses of irradiation (0.2 Gy) in mouse testis compared to the control group. However, testicular impairments were markedly evident in the group exposed to higher doses of carbon ions plus simulated microgravity, which may be due at least in part to elevated apoptosis initiated by the mitochondrial apoptosis pathway in germ cells.
分类: 生物学 >> 生物物理学 >> 生物物理、生物化学与分子生物学 提交时间: 2016-05-18
摘要: Autophagosome formation is promoted by the PI3 kinase complex and negatively regulated by myotubularin phosphatases, indicating that regulation of local phosphatidylinositol 3-phosphate (PtdIns3P) levels is important for this early phase of autophagy. Here, we show that the Caenorhabditis elegans myotubularin phosphatase MTM-3 catalyzes PtdIns3P turnover late in autophagy. MTM-3 acts downstream of the ATG-2/EPG-6 complex and upstream of EPG-5 to promote autophagosome maturation into autolysosomes. MTM-3 is recruited to autophagosomes by PtdIns3P, and loss of MTM-3 causes increased autophagic association of ATG-18 in a PtdIns3P-dependent manner. Our data reveal critical roles of PtdIns3P turnover in autophagosome maturation and/or autolysosome formation.
分类: 生物学 >> 生物物理学 >> 生物物理、生物化学与分子生物学 提交时间: 2016-05-12
摘要: UBXD8 is a membrane protein that mediates endoplasmic reticulum-associated protein ubiquitination and degradation by interacting with p97NCP. Recently, lipid droplet proteomic studies show the lipid droplet localization of UBXD8. Besides, UBXD8 is also involved in triglyceride metabolism. However, the molecular mechanism by which UBXD8 regulates triglyceride metabolism is still obscure. Here we knocked out UBXD8 in mouse C2C12 myoblasts by CRISPR/Cas9. We selected 2 UBXD8 knockout (KO) clone cell lines from 26 possible KO clones. UBXD8 KO did not change the lipid droplet proteins expression pattern. However, UBXD8 KO led to the accumulation of neutral lipid. Furthermore, our data show that UBXD8 KO could alleviate palmitate-induced insulin resistance and rescue palmitate-induced apoptosis which was characterized by PARP splicing. In addition, the phenotype of palmitate-induced insulin resistance and apoptosis was reappeared after overexpressing UBXD8 in UBXD8 KO cells. These data suggested that UBXD8 plays an important role in lipid metabolism and its abnormity related insulin signal and apoptosis.
分类: 生物学 >> 生物物理学 >> 生物物理、生物化学与分子生物学 提交时间: 2016-05-12
摘要: MutM (Formamidopyrimidine-DNA glycosylase, Fpg), a bifunctional base excision repair enzyme (DNA glycosylase/AP lyase), is involved in the repair of many kinds of DNA damage, including the formation of 8-oxoguanine, 5-formyluracil, and C/C mismatches, through recognizing DNA damage and removing damaged bases. The mechanisms of MutM involvement, however, with the exception of 8-oxoG, are poorly understood. Here, we identified proteins which interact with MutM in Mycobacterium smegmatis using methods of tandem affinity purification and mass spectrometry and used Far-western and GST pull-down analysis to validate the interactions between MutM and DEAD-box rna helicase, RpsC, and UvrA. Results demonstrated that tandem affinity purification is a suitable method for identifying MutM interacting proteins and provided insights into the mechanism by which MutM is involved in DNA damage repair.
分类: 生物学 >> 生物物理学 提交时间: 2016-05-12
摘要: Objective: Sera of Hashimoto's thyroiditis (HT) patients are known to exhibit elevated levels of anti-thyroglobulin IgG (TgAb IgG). Therefore, TgAb IgG represents a hallmark of this debilitating autoimmune disease. The aim of our study was to investigate the differential expression of specific glycosylation patterns of TgAb IgG from HT patients and healthy blood donors. Methods: HT patients (n = 32) were divided into two subgroups, medium level group (mHT, n = 15) and high level group (hHT, n = 17), according to the serum levels of TgAb detected by electrochemiluminescence immunoassay. TgAb IgG was purified by affinity chromatography from the sera of the HT group and control group (n = 15). MALDI-QIT-TOF-MS/MS spectrometry was performed to identify the glycosylation profiles of purified TgAb IgG. Lectin microarray technology was used to compare the abundance of different glycans found on TgAb IgG between HT patients and controls, and between the mHT and hHT groups. Results: The results by MALDI-QIT-TOF-MS/MS showed that the glycosylation profiles of TgAb IgG were similar between the mHT, hHT, and control groups. Furthermore, the lectin microarray showed that compared to the control group (all P < .001), there were higher levels present of (1) mannose (detected as lectin LCA, VFA, and MNA-M); (2) terminal sialic acid (detected as SNA-I and PSA); (3) core fucose (detected as LcH); and (4) Gal(beta 1-4) GlcNAc(beta 1-2) Man glycans (detected as PHA-L) on TgAb IgG from the HT group. A similar trend was observed between the hHT and mHT group, with elevated levels of mannose, terminal sialic acid, core fucose, and Gal(beta 1-4) GlcNAc(< 12) Man glycans on TgAb IgG found in the hHT group compared with the mHT group (all P < .05). Conclusions: TgAb IgG of HT patients exhibits higher glycosylation levels than those observed for TgAb IgG of healthy controls. Our results provide new clues for exploring the role of TgAb in the pathogenesis of HT.
提交时间: 2017-05-02
摘要: We describe the lateral-coupled junctionless indium-zinc-oxide [IZO] thin-film transistors [TFTs] in which there are no junctions between channel and source/drain electrodes and with solid-state phosphosilicate glass electrolyte [PSG] gating. Due to the t