Physiological responses to chilling in γ‐ray‐induced mutant 22‐3 compared with two wild‐type lines (W1 and W2) with yellow‐stemmed stolons from the cultivar Common were analyzed to understand the chilling tolerance mechanism of centipedegrass [Eremochloa ophiuroides (Munro) Hack.]. The mutant 22‐3 showed an elevated chilling tolerance, with lower ion leakage and higher chlorophyll content after chilling treatment than the wild‐type plants. The maximum photochemical efficiency, quantum efficiency of photosystem II photochemistry, photochemical quenching efficiency, and net photosynthetic rate decreased after chilling treatment but were less affected in 22‐3 than in the wild‐type plants. Consistently higher levels of maximum net photosynthetic rate, apparent quantum yield, carboxylation efficiency, and light saturation point and lower levels of the CO2 compensation point and light compensation point were observed in 22‐3 than in the wild‐type plants after chilling treatment. Antioxidant enzyme activities and polyamines (PAs) including putrescine (Put), spermidine (Spd), and spermine (Spm) remained higher in 22‐3 relative to the wild‐type plants during chilling stress. Activities of superoxide dismutase, ascorbate‐peroxidase (APX), catalase (CAT), and glutathione reductase (GR) were highly correlated with Put and Spm, while APX, CAT, and GR were highly correlated with Spd. The results suggest that higher levels of PAs and antioxidants are associated with the elevated chilling tolerance in 22‐3 and improve protection of photosynthesis against chilling‐induced oxidative damage. We propose that manipulation of the biosynthesis of PAs could be valuable in centipedegrass breeding for increased chilling tolerance.
BackgroundEnzyme-linked immunosorbent assay (ELISA) is a common method for diagnosing swine influenza. However, the production of classical antibodies is both costly and time-consuming. As a promising alternative diagnostic tool, single-domain antibodies (sdAbs) offer the advantages of simpler and faster generation, good stability and solubility, and high affinity and specificity.MethodsPhage display technology was used to isolate sdAbs against the SIV-NP protein from a camel VHH library. The sdAb5 was fused to the biotin acceptor peptide (BAP) and a His-Tag for its expression as monomeric and site-specific biotinylation in E.coli to develop an sdAb-based blocking ELISA (sdAb-ELISA). In the sdAb-ELISA, the anti-SIV antibodies from swine samples were used to block the binding between the biotinylated sdAb5 and SIV-NP protein coated on the ELISA plate. The specificity, sensitivity, and reproducibility of sdAb-ELISA were determined. In addition, consistency among sdAb-ELISA, commercial ELISA kit, and Western blot was evaluated.ResultsSix SIV-NP-specific sdAbs were isolated, among which sdAb5 was identified as a dominant sdAb with higher reactivity. The cut-off value of biotinylated sdAb5-based bELISA was determined to be 29.8%. Compared with the positive reference serum against five different types of swine viruses, the developed sdAb-ELISA showed 100% specificity. The detection limit of sdAb-ELISA was 1:160 in an anti-SIV positive reference serum, which is lower than that of the commercial ELISA kit (1:20). In 78 diluted anti-SIV positive serum (1:80), 21 and 42 samples were confirmed as positive by the commercial ELISA kit and sdAb-ELISA, respectively. The coefficients of variation of intra- and inter-assay were 1.79–4.57% and 5.54–9.98%, respectively. The sdAb-ELISA and commercial ELISA kit showed a consistency of 94.17% in clinical swine serum samples. Furthermore, the coincidence rate was 96.67% between the results detected by sdAb-ELISA and Western blot.ConclusionA specific, sensitive, and reproducible sdAb-ELISA was successfully developed, which offers a new, promising method to detect anti-SIV antibodies in swine serum.
Background The relationship between malnutrition-inflammation-atherosclerosis syndrome (MIAS) and self-management ability has not been previously revealed even though both play an important role in the management of peritoneal dialysis (PD) patients. Methods In total, 93 patients were enrolled in this study. A self-management questionnaire was used for the evaluation of self-management ability. The identification of MIAS was based on one or more of the following three conditions: C-reactive protein (CRP)≥10 mg/L, malnutrition–inflammation score (MIS)> 7, and the presence of atherosclerosis-related medical records. The possible association between different self-management abilities and MIAS was analyzed with a Spearman correlation analysis. Results There were 40 (43.0%) patients in the atherosclerosis group, and 38 (40.9%), 38 (40.9%), 10 (10.8%), and 7 (7.5%) patients in the MIAS0, MIAS1, MIAS2, and MIAS3 groups, respectively. The group with a score above the mean score of the Dialysis Effect Evaluation and Monitoring dimension had a fewer number of hospitalizations, higher albumin levels, lower MIS scores, a lower level of IL-6, and a lower number of MIAS factors. The Pearson and Spearman correlation analyses also revealed that this dimension was negatively correlated with the MIAS, MIS, IL-6, BNP, number of hospitalizations, and age and positively associated with albumin and prealbumin. Conclusion The Dialysis Effect Evaluation and Monitoring dimension of the self-management scale for PD patients is closely linked to the MIAS, and a better dialysis effect evaluation and monitoring capacity results in a decreased likelihood of exposure to malnutrition and inflammation. Trial registration Chinese Clinical Trial Registry: ChiCTR2000035525 (http://www.chictr.org.cn/showproj.aspx?proj=58110), registered August 13, 2020.
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