Single-step equipment-free extracellular vesicle concentration using super absorbent polymer beads.
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ABSTRACT:
SUBMITTER: Yang HC
PROVIDER: S-EPMC7902527 | biostudies-literature |
REPOSITORIES: biostudies-literature
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Project description:Detection and quantification of pathogens in water is critical for the protection of human health and for drinking water safety and security. When the pathogen concentrations are low, large sample volumes (several liters) are needed to achieve reliable quantitative results. However, most microbial identification methods utilize relatively small sample volumes. As a consequence, a concentration step is often required to detect pathogens in natural waters. Herein, we introduce a novel water sample concentration method based on superabsorbent polymer (SAP) beads. When SAP beads swell with water, small molecules can be sorbed within the beads, but larger particles are excluded and, thus, concentrated in the residual non-sorbed water. To illustrate this approach, millimeter-sized poly(acrylamide-co-itaconic acid) (P(AM-co-IA)) beads are synthesized and successfully applied to concentrate water samples containing two model microorganisms: Escherichia coli and bacteriophage MS2. Experimental results indicate that the size of the water channel within water swollen P(AM-co-IA) hydrogel beads is on the order of several nanometers. The millimeter size coupled with a negative surface charge of the beads are shown to be critical in order to achieve high levels of concentration. This new concentration procedure is very fast, effective, scalable, and low-cost with no need for complex instrumentation.
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Project description:Recently, there has been a high expectation that high water absorbent hydrogels can be developed as an artificial vitreous body. However, the drawbacks associated with in vivo instability, biofouling, uncontrollable in situ reaction time, and injection-induced precrosslinked fragmentation preclude their genuine use as vitreous substitutes. Here, a supramolecular binary copolymer hydrogel termed as PNAGA-PCBAA by copolymerization of N-acryloyl glycinamide (NAGA) and carboxybetaine acrylamide (CBAA) is prepared. This PNAGA-PCBAA hydrogel physically crosslinked by dual amide hydrogen bonds of NAGA exhibits an ultralow solid content (1.6, 98.4 wt% water content), and shear-thinning behavior, body temperature extrudability/self-healability, rapid network recoverability, and very close key parameters (modulus, antifouling/antifibrosis, light transmittance, refractive index, ultrastability) to human vitreous body. It is demonstrated that the hydrogel can be readily injected by a 22G needle into the rabbits' eyes where the gelling network is rapidly recovered. After 16 weeks postoperation, the hydrogel acts as a very stable vitreous substitute without affecting the structure of soft tissues in eye, or eliciting adverse effects. This supramolecular binary copolymer hydrogel finds a broad application in ophthalmic fields as not only a self-recoverable permanent vitreous substitute, but also transient intraocular filling for prevention of inner tissues in postsurgical eyes.
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Project description:Type 2 diabetes is a chronic age-associated degenerative metabolic disease that reflects relative insulin deficiency and resistance. Extracellular vesicles (EVs) (exosomes, microvesicles, and apoptotic bodies) are small (30-400 nm) lipid-bound vesicles capable of shuttling functional proteins, nucleic acids, and lipids as part of intercellular communication systems. Recent studies in mouse models and in cell culture suggest that EVs may modulate insulin signaling. Here, we designed cross-sectional and longitudinal cohorts of euglycemic participants and participants with prediabetes or diabetes. Individuals with diabetes had significantly higher levels of EVs in their circulation than euglycemic control participants. Using a cell-specific EV assay, we identified that levels of erythrocyte-derived EVs are higher with diabetes. We found that insulin resistance increases EV secretion. Furthermore, the levels of insulin signaling proteins were altered in EVs from individuals with high levels of insulin resistance and β-cell dysfunction. Moreover, EVs from individuals with diabetes were preferentially internalized by circulating leukocytes. Cytokine levels in the media and in EVs were higher from monocytes incubated with diabetic EVs. Microarray of these leukocytes revealed altered gene expression pathways related to cell survival, oxidative stress, and immune function. Collectively, these results suggest that insulin resistance increases the secretion of EVs, which are preferentially internalized by leukocytes, and alters leukocyte function.
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Project description:Biofertilizer is a good substitute for chemical fertilizer in sustainable agriculture, but its effects are often hindered by drought stress. Super absorbent polymer (SAP), showing good capacity of water absorption and retention, can increase soil moisture. However, limited information is available about the efficiency of biofertilizer amended with SAP. This study was conducted to investigate the effects of synergistic application of SAP and biofertilizers (Paenibacillus beijingensis BJ-18 and Bacillus sp. L-56) on plant growth, including wheat and cucumber. Potted soil was treated with different fertilizer combinations (SAP, BJ-18 biofertilizer, L-56 biofertilizer, BJ-18 + SAP, L-56 + SAP), and pot experiment was carried out to explore its effects on viability of inoculants, seed germination rate, plant physiological and biochemical parameters, and expression pattern of stress-related genes under drought condition. At day 29 after sowing, the highest viability of strain P. beijingensis BJ-18 (264 copies ng-1 gDNA) was observed in BJ-18 + SAP treatment group of wheat rhizosphere soil, while that of strain Bacillus sp. L-56 (331 copies ng-1 gDNA) was observed in the L-56 + SAP treatment group of cucumber rhizosphere soil. In addition, both biofertilizers amended with SAP could promote germination rate of seeds (wheat and cucumber), plant growth, soil fertility (urease, sucrose, and dehydrogenase activities). Quantitative real-time PCR analysis showed that biofertilizer + SAP significantly down-regulated the expression levels of genes involved in ROS scavenging (TaCAT, CsCAT, TaAPX, and CsAPX2), ethylene biosynthesis (TaACO2, CsACO1, and CsACS1), stress response (TaDHN3, TaLEA, and CsLEA11), salicylic acid (TaPR1-1a and CsPR1-1a), and transcription activation (TaNAC2D and CsNAC35) in plants under drought stress. These results suggest that SAP addition in biofertilizer is a good tactic for enhancing the efficiency of biofertilizer, which is beneficial for plants in response to drought stress. To the best of our knowledge, this is the first report about the effect of synergistic use of biofertilizer and SAP on plant growth under drought stress.
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Project description:One of the major factors limiting the production of medicinal plants in arid and semi-arid areas is water deficit or drought stress. One-third of the land in the world is arid and semi-arid and is inhabited by nearly 4 × 108 people. Ocimum basilicum (sweet basil) is a valuable medicinal plant that is sensitive to water deficit, and water shortage negatively affects sweet basil yield and quality. Water availability in the root zone of basil could ameliorate the negative effects of water shortage. To the best of our knowledge, although the effects of hydrophilic polymers (HPs) have been studied in different agricultural crops, the effects of HP application in medicinal plants have not been previously investigated. This investigation was conducted to explore the effects on water use efficiency when using Stockosorb® (STS) and psyllium seed mucilage (PSM) as hydrophilic polymers (HPs) and the effects of these HPs on essential oil quality, quantity, and yield. The research was set up in a factorial experiment on the basis of completely randomized block design with three replications. We used two HPs, STS (industrial) and PSM (herbal), with two methods of application (mixed with soil, mixed with soil + root) at four concentrations (0%, 0.1%, 0.2%, and 0.3% (w/w)). Results showed that the STS and PSM significantly increased the dry herb yield (both shoot and root) in comparison to the control, and the improving effect was higher when these HPs were mixed with soil + root. The highest dry herb yield (6.74 and 3.68 g/plant for shoot and root, respectively) was detected in the PSM at 0.1% mixed with soil + root. There was not any significant difference in dry herb yield among PSM (0.1%), PSM (0.2%), and STS (0.2%) when mixed with soil + root. Soil application of PSM and soil + root application of STS at a concentration of 0.3% increased the Essential Oil (EO) content almost three-fold in comparison to the control (0.5% and 0.52% to 0.18% v/w, respectively). The maximum essential oil yield was recorded in plants treated with STS (0.2% in) or PSM (0.1%) by soil + root application (0.21 and 0.19 mL/plant, respectively). PSM at concentrations of 0.1% and 0.2% (mixed with soil + root) showed the highest water use efficiency (1.91 and 1.82 g dry weight (DW)/L H2O, respectively). STS mixed with soil also significantly improved water use efficiency (WUE) in comparison to the control. The application of these HPs improved the quality of sweet basil essential oil by increasing the linalool and decreasing the eugenol, epi-?-cadinol, and trans-?-bergamotene content.
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Project description:BackgroundRecently, bacterial extracellular vesicles (EVs) have been considered to play crucial roles in various biological processes and have great potential for developing cancer therapeutics and biomedicine. However, studies on bacterial EVs have mainly focused on outer membrane vesicles released from gram-negative bacteria since the outermost peptidoglycan layer in gram-positive bacteria is thought to preclude the release of EVs as a physical barrier.ResultsHere, we examined the ultrastructural organization of the EV produced by gram-positive bacteria using super-resolution stochastic optical reconstruction microscopy (STORM) at the nanoscale, which has not been resolved using conventional microscopy. Based on the super-resolution images of EVs, we propose three major mechanisms of EV biogenesis, i.e., membrane blebbing (mechanisms 1 and 2) or explosive cell lysis (mechanism 3), which are different from the mechanisms in gram-negative bacteria, despite some similarities.ConclusionsThese findings highlight the significant role of cell wall degradation in regulating various mechanisms of EV biogenesis and call for a reassessment of previously unresolved EV biogenesis in gram-positive bacteria.
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Project description:The aim of this study was to determine the effectiveness of a Super absorbent polymer (SAP) containing copper (SAP-Cu) in controlling mal secco disease (MSD) of lemon caused by the fungus Plenodomus tracheiphilus. Super absorbent polymer containing copper was characterized by atomic absorption spectrometry (AAS) and UV-VIS spectroscopy. In vitro tests were performed to determine the inhibitory effects of SAP-Cu against the pathogen on both potato-dextrose-agar medium and naturally infected lemon cuttings. Super absorbent polymer was able to absorb up to about 200 and 30 times its weight of ionized water and copper (II) sulfate solution (Cu2+ ions at the concentration 236 mM), respectively. The distribution of copper released on twigs after 24 h of contact with SAP-Cu was determined by secondary ion mass spectrometry with time-of-flight analyzer (ToF-SIMS). Super absorbent polymer containing copper significantly inhibited the viability of P. tracheiphilus in lemon twigs. Overall, the results of this study showed that the SAP could be a suitable carrier of antifungal compounds.
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Project description:Red blood cell (RBC) hemolysis is one of the most common storage lesions in packed RBCs (pRBC). Older units of pRBCs, especially those?>?21 days old, have increasing levels of hemolysis leading to increased oxidative stress and premature platelet activation. This effect can mostly be attributed to the increase of cell-free hemoglobin (Hb). Therefore, removal of cell-free Hb from pRBCs prior to transfusion could mitigate these deleterious effects. We propose a new method for the removal of Hb from pRBCs using zinc beads. Prepared Hb solutions and pRBCs were treated with zinc beads using two different protocols. UV-Vis spectrophotometry was used to determine Hb concentrations, before and after treatment. Experiments were run in triplicate and paired t tests were used to determine significant differences between groups. Zinc beads removed on average 94% of cell-free Hb within 15 min and 78% Hb from pRBCs (p?<?0.0001), demonstrating a maximum binding capacity?~?66.2?±?0.7 mg Hb/mL beads. No differences in RBC morphology or deformability were observed after treatment. This study demonstrates the feasibility of using zinc beads for the rapid and targeted removal of Hb from pRBC units. Further investigation is needed to scale this method for large volume removal.
| S-EPMC6506348 | biostudies-literature
Project description:The main forms of poly-γ-glutamic acid (γ-PGA) applied in agriculture include agricultural γ-PGA and γ-PGA super absorbent polymer (SAP). Laboratory experiments were conducted with a check treatment CK (no γ-PGA added) and two different forms of γ-PGA added to sandy loam soil (T and TM stand for γ-PGA and γ-PGA SAP) at four different soil mass ratios (0.05% (1), 0.10% (2), 0.15% (3) and 0.20% (4)) to determine their effects on sandy loam soil hydro-physical properties. Both of them could reduce the cumulative infiltration of soil water. The total available water (TAW) which the soil water content (SWC) from field water capacity (FC) to permanent wilting point (PWP) after γ-PGA added into sandy loam soil had no significant different compared with CK, and the TAW was highest at the treatment of γ-PGA with 0.10% addition amount into sandy loam soil. However, the TAW of sandy loam soil increased dramatically with the γ-PGA SAP addition amount increasing. TM3 had the highest soil water absorption among the treatments with γ-PGA SAP. The T1 to T4 treatments with γ-PGA addition slightly prolonged retention time (RT) when SWC varied from FC to PWP compared with CK. For γ-PGA SAP addition treatments, the time for SWC varied from FC to PWP was 1.48 times (TM1), 1.88 times (TM2), 2.01 times (TM3) and 2.87 times (TM4) longer than that of CK, respectively. The results of this study will provide further information for the use of these materials in agricultural application.
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Project description:ABSTRACTThe microRNA (miRNA) cargo contained in plasma extracellular vesicles (EVs) offers a relatively little explored source of biomarkers for brain diseases that can be obtained noninvasively. Methods to isolate EVs from plasma, however, are still being developed. For EV isolation, it is important to ensure the removal of vesicle-free miRNAs, which account for approximately two-thirds of plasma miRNAs. Membrane particle precipitation-based EV isolation is an appealing method because of the simple protocol and high yield. Here, we evaluated the performance of a precipitation-based method to obtain enriched EV-specific miRNAs from a small volume of rat plasma. We performed size-exclusion chromatography (SEC) on precipitation-isolated EV pellets and whole plasma. The SEC fractions were analysed using Nanoparticle Tracking Analysis (NTA), protein and miRNA concentration assays, and droplet digital polymerase chain reaction for four miRNAs (miR-142-3p, miR-124-3p, miR-23a, miR-122). Precipitation-isolated EVs and selected SEC fractions from the plasma were also analysed with transmission electron microscopy (TEM). Precipitation-based EV isolation co-precipitated 9% to 15% of plasma proteins and 21% to 99% of vesicle-free miRNAs, depending on the individual miRNAs. In addition, the amount of miR-142-3p, found mainly in EV fractions, was decreased in the EV fractions, indicating that part of it was lost during precipitation-based isolation. Western blot and TEM revealed both protein and lipoprotein contamination in the precipitation-isolated EV-pellets. Our findings indicate that a precipitation-based method is not sufficient for purifying plasma EV-contained miRNA cargo. The particle number measured by NTA is high, but this is mostly due to the contaminating lipoproteins. Although a part of the vesicle-free miRNA is removed, vesicle-free miRNA still dominates in plasma EV pellets isolated by the precipitation-based method.
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