Additional file 1: of The advantages and limitations of guideline adaptation frameworks

Table S1. Factors influencing local guideline group’s decisions about how to construct recommendations for a new guideline. Table S2. Steps for the implementation of guidelines. (DOCX 31 kb

Mitochondria are the major sources of ROS in hyperthermia-treated platelets.

<p>(A and B) DCFDA-loaded platelets were incubated at the indicated temperatures for 3 h (A) or 1-3 h (B). As a positive control, loaded platelets were incubated with thrombin at 37°C for 30 min. Samples were then analyzed for intracellular ROS levels by flow cytometry. Representative flow cytometric histogram is shown (A). The relative ROS levels are expressed as a percentage of platelets, which were incubated at RT for 1 h (B). Percentage of RT 1h is presented as mean ± SEM from three independent experiments. *<i>P</i><0.017 (after Bonferroni correction) as compared with RT 1 h, **<i>P</i><0.017 (after Bonferroni correction) as compared with RT 2 h, #<i>P</i><0.017 (after Bonferroni correction) as compared with RT 3h. (C–E) DCFDA-loaded platelets were pre-incubated with solvent control, DPI and apocynin (C), Mito-TEMPO and NAC (D), or L-NAME and ETYA (E) at 37 °C for 15 min, and then incubated for 3 h at 40°C or 42°C, and further analyzed by flow cytometry. The relative ROS levels are expressed as a percentage of platelets, which were pre-incubated with solvent control at 37°C for 15 min and then incubated at 40°C for 3 h. Percentage of 40°C loaded platelets pre-incubated with solvent control is presented as mean ± SEM from three independent experiments. *<i>P</i><0.025 (after Bonferroni correction) as compared with solvent control at 40°C, **<i>P</i><0.025 (after Bonferroni correction) as compared with solvent control at 42°C. Thrombin is labeled as Thr.</p

Hyperthermia increase mitochondrial superoxide production.

<p>(A and B) MitoSOX^TM Red-loaded platelets were incubated at the indicated temperatures for 3 h (A) or1-3 h (B). As a positive control, loaded platelets were incubated with antimycin A at 37°C for 30 min. Samples were then analyzed by flow cytometry. Representative flow cytometric histogram is shown (A). Data are expressed as a percentage of platelets which were incubated at RT for 1h (B). Percentage of RT 1h is presented as mean ± SEM from three independent experiments. *<i>P</i><0.017 (after a Bonferroni correction) compared with RT 1 h, **<i>P</i><0.017 (after Bonferroni correction) as compared with RT 2 h, #<i>P</i><0.017 (after Bonferroni correction) as compared with RT 3 h. (C) MitoSOX^TM Red-loaded platelets were pre-incubated with apocynin, Mito-TEMPO, L-NAME, ETYA, or solvent control at 37°C for 15 min, and then incubated for 3 h at 40°C or 42°C, and further analyzed by flow cytometry. Data are expressed as a percentage of platelets that were pre-incubated with solvent control at 37°C for 15 min and then incubated at 40°C for 3 h. Percentage of 40°C platelets pre-incubated with solvent control is presented as mean ± SEM from three independent experiments. *<i>P</i><0.013 (after Bonferroni correction) as compared with solvent control at 40°C, **<i>P</i><0.013 (after Bonferroni correction) as compared with solvent control at 42°C. Antimycin A is labeled as Ant.</p

Hyperthermia decreases MnSOD protein levels and enzyme activity.

<p>(A and B) Washed platelets were incubated at the indicated temperatures for 3 h. Treated platelets were lysed, and ManSOD activity was measured using a commercially available SOD (A), or GPx4 (B) assay kits. *<i>P</i><0.017 (after Bonferroni correction) as compared with RT. (C and D) Washed platelets were incubated at the indicated temperatures for 3 h. Treated platelets were lysed and analyzed by Western blot with anti-MnSOD (C), or anti-GPx4 (D). Actin levels demonstrated equal protein loading. Representative results of three independent experiments are presented.</p

Effect of Mito-TEMPO on caspase-3 activation, depolarization of ΔΨm, and PS exposure in hyperthermia-treated platelets.

<p>(A and B) Washed platelets were incubated at indicated temperatures for 3 h (A) or pre-incubated with Mito-TEMPO and solvent control at 37°C for 15 min and then incubated at 42°C for 3 h (B). Treated platelets were lysed and analyzed by Western blot with anti-cleaved p17 fragment of caspase-3. Actin levels were assayed to demonstrate equal protein loading. Representative results of three independent experiments are presented. (C–F) Washed platelets were pre-incubated without (C and D) or with Mito-TEMPO and solvent control at 37°C for 15 min (E and F), and then incubated at the indicated temperature for 3 h. Treated platelets were incubated with TMRE (C and E), or annexin V-FITC (D and F), and analyzed by flow cytometry. Representative flow cytometric histograms are shown (C and D). Data are expressed as a percentage of platelets that were pre-incubated with solvent control at 37°C for 15 min and then incubated at RT for 3 h (E and F). The percentage of RT platelets pre-incubated with solvent is presented as mean ± SEM from three independent experiments. *<i>P</i><0.05 as compared with solvent control at an identical temperature.</p

Effect of Mito-TEMPO on Bax mitochondrial translocation and cytochrome C release in hyperthermia-treated platelets.

<p>(A–D) Washed platelets were incubated at different temperatures for 3 h (A and B), or pre-incubated with Mito-TEMPO and solvent control at 37°C for 15 min and then incubated at 42°C for 3 h (C and D). Treated platelets were lysed, and cytosol and mitochondrial fractions were isolated and analyzed by Western blot with anti-Bax (A and C), and anti-cytochrome C (B and D). Cytochrome C oxidase subunit 1 (COX1) and tubulin were used as internal controls. Representative data of three independent experiments are presented. Cytochrome C is labeled as Cyto C.</p

Mito-TEMPO ameliorates hyperthermia-impaired platelet function.

<p>(A) Platelet aggregation was performed as described in Methods. Representative traces from three independent experiments are shown. (B and C) Washed platelets were incubated at the indicated temperatures for 3 h (B), or pre-incubated with Mito-TEMPO, GM6001 and solvent control at 37°C for 15 min, followed by incubation at 42°C for 3 h (C). Western blot was performed as described in Methods. (D and E) Washed platelets were incubated at indicated temperatures for 3 h (D), or pre-incubated without (control) or with Mito-TEMPO and solvent at 37°C for 15 min, and then incubated at 42°C for 3 h (E). Platelet adhesion was performed as described in Methods. The results shown are the mean ± SEM of cell number/mm². *<i>P</i><0.017 (after Bonferroni correction) as compared with RT, **<i>P</i><0.025 (after Bonferroni correction) as compared with control. Mito-TEMPO is labeled as TEMPO.</p

Effect of Mito-TEMPO on the levels of MDA and cardiolipin peoxidation in hyperthermia-treated platelets.

<p>(A and B) Washed platelets were incubated without (A), or with (B) Mito-TEMPO and solvent control at 37°C for 15 min, and then incubated at different temperatures for 3 h. MDA levels were detected as described in Methods. (C–E) Washed platelets were incubated without (C and D), or with (E) Mito-TEMPO and solvent control at 37°C for 15 min, and then incubated at different temperatures for 3 h. Cardiolipin peoxidation was detected as described in Methods. Representative flow cytometric histogram is shown (C). (A and D) Data are expressed as a percentage of platelets which were incubated at RT for 3h. Percentage of RT is presented as mean ± SEM from three independent experiments. *<i>P</i><0.017 (after Bonferroni correction) as compared with RT. (B and E) Data are expressed as a percentage of platelets that were pre-incubated with solvent control at 37°C for 15 min and then incubated at RT for 3 h. Percentage of RT platelets pre-incubated with solvent control is presented as mean ± SEM from three independent experiments. *<i>P</i><0.05 as compared with solvent control at an identical temperature.</p

Replacing Phenol and Formaldehyde for Green Adhesive Synthesis by Lignin and Furfural

Traditional phenol-formaldehyde resin involves the use of petrochemical-based phenol and formaldehyde as feedstocks, raising environmental and health concerns. Herein, renewable biobased substances, i.e., lignin and furfural, were employed to partially replace petroleum-based phenol and formaldehyde for developing greener adhesives by batch copolymerization method. The substitution rates of phenol by acetonitrile-extracted lignin and formaldehyde by furfural can reach 80 and 15%, respectively. The application of these greener raw materials can effectively decrease the emission of toxic content, increasing the viscosity and thermal stability without any sacrifice in the binding performance. The resin with 40% phenol substitution rate and 15% formaldehyde substitution rate displayed superior trade-off between bonding strength (a wet tensile strength of 1.68 MPa) and free formaldehyde content (0.21%) among all biobased resins, much superior to those of commercial adhesive (0.7 MPa and 5%) and those prepared with industrial lignin (0.66 MPa and 0.38%). By using two major biomass-derived feedstocks, this work provides a sustainable strategy to develop a green adhesive featuring superior adhesive properties, enhanced environmental compatibility, and improved thermal stability that shows a broader application potential in the adhesive field

Liposomal Resveratrol Alleviates Platelet Storage Lesion via Antioxidation and the Physical Buffering Effect

Platelet transfusion is essential in the treatment of platelet-related diseases and the prevention of bleeding in patients with surgical procedures. Platelet transfusion efficacy and shelf life are limited mainly by the development of platelet storage lesion (PSL). Mitigating PSL is the key to prolonging the platelet shelf life and reducing wastage. Excess intracellular reactive oxygen species (ROS) are one of the main factors causing PSL. In this study, we explored a nanomedicine strategy to improve the quality and functions of platelets in storage. Resveratrol (Res), a natural plant product, is known for its antioxidative effect. However, medical applications of Res are limited due to its low water solubility and stability. Therefore, we used a resveratrol-loaded liposomal system (Res-Lipo) to better utilize the antioxidant effect of the drug. This study aimed to evaluate the effect of Res-Lipo on platelet oxidative stress and alleviation of PSL during the storage time. Res-Lipo scavenged intracellular ROS and inhibited platelet apoptosis and activation during storage. Res-Lipo not only maintained mitochondrial function but also improved platelet aggregation in response to adenosine 5′-diphosphate. These results revealed that Res-Lipo ameliorated PSL and prolonged the platelet survival time in vivo. The strategy provides a potential method for extending the platelet storage time and might be considered a potential and safe additive to alleviate PSL