25 research outputs found

    Structure and pH-Induced Structural Rearrangements of the Putative Multidrug Efflux Pump EmrD in Liposomes Probed by Site-Directed Spin Labeling

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    EmrD is the only structurally characterized drug/H<sup>+</sup> antiporter of the major facilitator superfamily (MFS). It has been crystallized in a doubly occluded conformation that is considered representative of an intermediate state in the transport cycle of MFS transporters. However, unexpected features of the crystal structure and the lack of functional information available for EmrD limit the utility of the structural data. To assess whether the crystal structure represents a stable state in a native-like environment, we used electron paramagnetic resonance (EPR) spectroscopy to determine the mobility and accessibility of spin labels at 76 positions in six transmembrane (TM) helices of EmrD reconstituted in liposomes. While the EPR data were mostly consistent with the crystal structure, they also revealed significant deviations from the predicted orientation and topology of TM helices at several locations. Additionally, we were unable to reproduce EmrD-dependent multidrug resistance phenotypes <i>in vitro</i> and in cell-based assays of drug transport. In spite of structural and functional discrepancies, we mapped a pH-dependent conformational change in which the cytoplasmic side of the N-terminal half opened locally in response to protonation. This conformational switch is consistent with the expected pH-dependent behavior of MFS H<sup>+</sup>-coupled antiporters

    This is the GRAMMS checklist.

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    BackgroundCoronary Heart Disease (CHD) is one of the most prevalent chronic diseases worldwide. Currently, percutaneous coronary intervention (PCI) with stent implantation is the main clinical treatment for CHD, and patients can achieve better outcomes after stenting. However, adverse cardiovascular events continue to recur, ultimately failing to yield good results. Several symptoms exist after stenting and are associated with health outcomes. Little is known about the symptom patterns of patients during the different postoperative periods. Therefore, this study aims to explore the dynamics of symptoms and clarify the experiences of post-stenting in patients during different periods, which may help the delivery of more specific patient management and improve survival outcomes in the future.MethodsA mixed method (quantitative/qualitative) design will be adopted. Longitudinal research, including surveys regarding three different periods, will be sued to describe the symptom patterns of patients undergoing PCI with stent implantation, clarifying their focused symptom problems during different time periods or in populations with different features. Qualitative individual interviews aim to understand the feelings, experiences, opinions, and health conditions of patients post-stenting, which can explain and supplement quantitative data. Quantitative data will be analyzed using descriptive statistics, latent class analysis (LCA), and latent translation analysis (LTA). Qualitative data will be analyzed using content analysis.DiscussionThis study is the first study to explore the symptom patterns and experiences of patients in various domains after stent implantation using a novel design including quantitative and qualitative methods, which will help the delivery of more specific patient management, reduce the recurrence of adverse cardiovascular events, and improve survival outcomes in the future. It is also meaningful to use PROMIS profile-57 to help patients to proactively focus on their health problems, promote health literacy, and incorporate active patient participation into health management, which is a successful transition from passive medical treatment to active management.</div

    Effect of MDS-MSC on T cell apoptosis.

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    <p>T cells were incubated for 3 days alone or with MDS-MSC or normal-MSC in the presence of the mitogen PHA.The test was conducted by Annexin-V and PI double staining and analyzed by flow cytometry. Data are expressed as mean±SD of triplicates of 5 separate experiments. Annexin V+ means the cells were PI negative and Annexin V positive. *P≤0.05.</p

    MDS-MSC induce CD4+CD25+Foxp3+Tregs.

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    <p>(A) CD4+CD25-T cells were cultured with MDS-MSC or normal-MSC for 5 days, and CD4+ T cells were collected. The expression of CD25 and Foxp3 on CD4+ T cells was analyzed by FACS. Results are expressed as mean±SD of triplicates of 4 separate experiments. *P≤0.05. (B) CD4+T cells were cocultured with MDS-MSC generated CD4+CD25+Foxp3+Tregs or normal-MSC generated CD4+CD25+Foxp3+Tregs in the presence of PHA, and the T-lymphocyte proliferation was measured on day 5 by [3H]-thymidine incorporation. Results are expressed as mean±SD of triplicates of 4 separate experiments. *P≤0.05. (C) MDS-MSC generated CD4+CD25+Foxp3+Tregs or normal-MSC generated CD4+CD25+Foxp3+Tregs inhibited the response of allogeneic T-lymphocyte in a dose-dependent manner. Responder CD2+ T-lymphocyte were stimulated with PHA for 5 days with or without graded dosed of MDS-MSC generated CD4+CD25+Foxp3+Tregs or normal-MSC generated CD4+CD25+Foxp3+Tregs. Results are expressed as mean±SD of triplicates of 4 separate experiments. *p≤0.05. (D) CD4+CD25-T cells were cultured with high-risk MDS-MSC or low-risk MDS-MSC for 5 days, and CD4+ T cells were collected. The expression of CD25 and Foxp3 on CD4+ T cells was analyzed by FACS. Results are expressed as mean±SD of triplicates of 4 separate experiments. *P≤0.05.</p

    Induction of CD4+CD25+Foxp3+Tregs by MDS-MSC is dependent on TGFβ1.

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    <p>Western blot confirmed efficient knockdown of TGF-β1. (B) CD4+CD25-T cells were cultured with TGF-β1 knockdown MDS-MSC or normal-MSC for 5 days, and CD4+ T cells were collected. The expression of CD25 and Foxp3 on CD4+ T cells was analyzed by FACS. Results are expressed as mean±SD of triplicates of 5 separate experiments. *P≤0.05. (C) CD4+CD25-T cells were cultured with mutant siRNA transfected MDS-MSC or normal-MSC for 5 days, and CD4+ T cells were collected. The expression of CD25 and Foxp3 on CD4+ T cells was analyzed by FACS. Results are expressed as mean±SD of triplicates of 6 separate experiments. *P≤0.05. (D) anti-rhTGF-β1 mAb was added at the beginning of coculture of CD4+CD25-T cells and untransfected MDS-MSC or normal-MSC for 5 days, and CD4+ T cells were collected. The expression of CD25 and Foxp3 on CD4+ T cells was analyzed by FACS. Results are expressed as mean±SD of triplicates of 6 separate experiments. *P≤0.05.</p

    MDS-MSC inhibit T-lymphocyte proliferation.

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    <p>Irradiated (15 Gy) MDS-MSC or normal-MSC were cultured for 5 days with CD2+ T-lymphocyte in the presence of PHA, then assessed by [3H]-thymidine incorporation. Data are expressed as mean±SD of triplicates of 5 separate experiments. *P≤0.05.</p

    Understanding adaptive tasks in cardiac rehabilitation among patients with acute myocardial infarction: a qualitative study

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    While Cardiac Rehabilitation (CR) programs have shown effectiveness in improving cardiac outcomes, there is limited understanding of how patients perceive and adapt to these interventions. Furthermore, alternative modes of delivering CR that have received positive evaluations from participants remain underexplored, yet they have the potential to enhance CR uptake. To explore the patient experience in CR programmes following Acute Myocardial Infarction (AMI) and describe their adaptive processing. This qualitative study was conducted at a nationally certified centre in China between July 2021 and September 2022, encompassing three stages: in-hospital, centre-based, and home-based CR programs. Purposive sampling was used to select eligible AMI patients for in-depth semi-structured interviews. The interview outline and analytical framework were aligned with the key concepts derived from the middle-range theory of adaptation to chronic illness and the normalization process theory. The findings were reported following the Consolidated Criteria for Reporting Qualitative Research checklist. Forty AMI patients were recruited. Four main themes describing the process of AMI patients normalizing CR intervention were identified, including (1) experiencing CR service driving by role’s responsibilities, (2) engaging in collaborative relationship based on interpersonal trust, (3) exploring a personalized rehabilitation plan by complex integration, and (4) expecting a promised outcome to shape decision-making. Integrated care interventions for AMI patients could benefit from a collaborative co-designed approach to ensure that CR interventions are normalized and fit into patients’ daily lives. Organizational-level CR services should align with the rehabilitation needs and expectations of patients.</p

    Effect of Sulfated Chitooligosaccharides on Wheat Seedlings (Triticum aestivum L.) under Salt Stress

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    In this study, sulfated chitooligosaccharide (SCOS) was applied to wheat seedlings to investigate its effect on the plants’ defense response under salt stress. The antioxidant enzyme activities, chlorophyll contents, and fluorescence characters of wheat seedlings were determined at a certain time. The results showed that treatment with exogenous SCOS could decrease the content of malondialdehyde, increase the chlorophyll contents, and modulate fluorescence characters in wheat seedlings under salt stress. In addition, SCOS was able to regulate the activities of antioxidant enzymes containing superoxide dismutase, catalase, peroxidase, ascorbate peroxidase, glutathione reductase, and dehydroascorbate reductase. Similarly, the mRNA expression levels of several antioxidant enzymes were efficiently modulated by SCOS. The results indicated that SCOS could alleviate the damage of salt stress by adjusting the antioxidant enzyme activities of plant. The effect of SCOS on the photochemical efficiency of wheat seedlings was associated with its enhanced capacity for antioxidant enzymes, which prevented structure degradation of the photosynthetic apparatus under NaCl stress. Furthermore, the effective activities of alleviating salt stress indicated the activities of SCOS were closely related with the sulfate group

    MDS-MSC inhibit the endocytosis of monocyte derived DCs.

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    <p>DCs <b>(5×10<sup>5</sup>)</b> were incubated in medium with FITC-labeled dextran at a concentration of 1 mg/ml in the presence of MSC <b>(5×10<sup>4</sup>)</b> at day 7. After an incubation period of 60 min at 37°C (green line) or 4°C as a control (gray line), cells were harvested and analyzed by FACS.Numbers in histograms indicate the mean fluorescence of each cell population. Results are expressed as mean±SD of triplicates of 5 separate experiments. *P≤0.05.</p

    Table_1_Low-Molecular-Weightt Polysaccharides From Pyropia yezoensis Enhance Tolerance of Wheat Seedlings (Triticum aestivum L.) to Salt Stress.DOCX

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    <p>Soil salinity is one of the major issues worldwide that affects plant growth and reduces agricultural productivity. Seaweed polysaccharides have been shown to promote crop growth and improve the resistance of plant to abiotic stresses. Pyropia yezoensis is a commercially important edible red alga in Southeast Asia. However, there is little research on the application of polysaccharides from P. yezoensis in agriculture. The molecular weight (MW) of polysaccharides influences their properties. Therefore, in this study, four representative polysaccharides from P. yezoensis (PP) with different MWs (MW: 3.2, 10.5, 29.0, and 48.8 kDa) were prepared by microwave-assisted acid hydrolysis. The relationship between the degradation of polysaccharides from P. yezoensis (DPP) and their effects on plant salt tolerance was investigated. The results showed that exogenous PP and DPPs increased wheat seedling shoot and root lengths, and fresh and dry weights, alleviated membrane lipid peroxidation, increased the chlorophyll content and enhanced antioxidant activities. The expression level examination analysis of several Na<sup>+</sup>/K<sup>+</sup> transporter genes suggested that DPPs could protect plants from the damage of salt stress by coordinating the efflux and compartmentation of Na<sup>+</sup>. The results demonstrated that polysaccharides could regulate antioxidant enzyme activities and modulate intracellular ion concentration, thereby to protect plants from salt stress damage. Furthermore, there was a significant correlation between the tolerance of wheat seedlings to salt stress and MW of polysaccharides. The results suggested that the lower-MW samples (DPP1, 3.2 kDa) most effectively protect wheat seedlings against salt stress.</p
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