Comparative investigation of exosome extraction from rat bone marrow mesenchymal stem cells using three different methodologies.

Exosomes have been identified as crucial mediators in numerous physiological and pathological processes, emerging as a focal point of scientific inquiry. This study aims to compare three methods for isolating exosomes from rat bone marrow mesenchymal stem cells: ultracentrifugation (UC), ultrafast separation system (EXODUS), and commercial precipitation kit (EXO-kit). First, the investigation compared exosomal morphology, particle size distribution, and expression of marker proteins. Subsequently, the RNA content, protein concentration, and purity of exosomes were evaluated. Finally, the impact of these exosomes on cellular metabolic viability and migration capacity was assessed. Results indicated that exosomes exhibited spherical or elliptical membrane structures, and most of the exosomes extracted by the three methods were in the range of 30to 200 nm. UC-extracted exosomes demonstrated the least impurities and clearest background, followed by EXODUS-extracted exosomes, and lastly EXO-kit-extracted exosomes. The EXO-kit-extracted exosomes yielded the highest RNA and protein content, whereas those isolated through UC exhibited superior purity. Furthermore, exosomes extracted from EXODUS and EXO-kit methods effectively enhanced the metabolic viability and migratory ability of osteoblast precursor cells compared to UC-extracted exosomes. In conclusion, each of the three methodologies presents advantages and limitations. Therefore, the selection of an appropriate exosome extraction technique should be based on specific experimental objectives and requirements.

Micro(nano)plastics: invisible compounds with a visible impact.

The plastic related research has been an epicentre in recent times. The presence and spread of micro (nano) plastics (MNPs) are well-known in the terrestrial and aquatic environment. However, the focus on the fate and remediation of MNP in soil and groundwater is limited. The fate and bioaccumulation of ingested MNPs remain unknown within the digestive tract of animals. There is also a significant knowledge gap in understanding the ubiquitous organic environmental pollutants with MNPs in biological systems. Reducing plastic consumption, improving waste management practices, and developing environmentally friendly alternatives are some of the key steps needed to address MNP pollution. For better handling and to protect the environment from these invisible substances, policymakers and researchers urgently need to monitor and map MNP contamination in soil and groundwater.

A simple overflow density separation method that recovers >95% of dense microplastics from sediment.

Density separation can isolate microplastics from environmental samples containing sediment. Typically, a solution added to sediment causes microplastics with lower densities to float. The solution of choice can influence the recovery of different particles since denser solutions can separate a greater range of microplastics. The equipment and procedural complexity further influence density separation protocols and microplastic recoveries. Zinc chloride (ZnCl2) is frequently used to isolate high-density polymers from environmental samples yet is rarely validated with simple, well-described protocols. A simple overflow method using ZnCl2 to isolate microplastics from sediment samples is described following a 3-step process: (1. Separation) ZnCl2 (1.7 g cm-3) solution is added to a sediment sample, agitated then settled; (2. Overflows) buoyant particles at the surface of the solution are overflowed twice; (3. Filtration) the overflowed solution is filtered. In a validation experiment with polyamide, rubber, polyvinyl chloride and polyethylene terephthalate/polyester, the mean recovery using this overflow method was 96 % ± 0.6 (standard error). This overflow density separation method proposes an accessible and reliable protocol to extract medium and high-density microplastics.•Microplastic separation with concentrated ZnCl2 solution•Simple overflow of buoyant particles•Reliable extraction of microplastics.

Parameter Identification of Multispan Rigid Frames Using a Stiffness Separation Method.

Identifying the parameters of multispan rigid frames is challenging because of their complex structures and large computational workloads. This paper presents a stiffness separation method for the static response parameter identification of multispan rigid frames. The stiffness separation method segments the global stiffness matrix of the overall structure into the stiffness matrices of its substructures, which are to be computed, thereby reducing the computational workload and improving the efficiency of parameter identification. Loads can be applied individually to each separate substructure, thereby guaranteeing obvious local static responses. The veracity and efficacy of the proposed methodology are substantiated by applying it to three- and eight-span continuous rigid frame structures. The findings indicate that the proposed approach significantly enhances the efficiency of parameter identification for multispan rigid frames.

A study on the application of blunt separation method in midline catheter intubation in elderly patients with coagulation dysfunction / 中华护理杂志

Objective To explore the efficacy of blunt separation method in midline catheter intubation in elderly patients with coagulation dysfunction.Methods A total of 80 elderly patients with coagulation dysfunction were selected in the convenience sampling method from October 2022 to April 2023 in our hospital,and they were randomly divided into an experimental(blunt)group and a control(routine)group,with 40 patients in each group.The differences in the degree of bleeding and exudation at the puncture site,the pain score and the incidence of complications(including bleeding and exudation,phlebitis,symptomatic catheter-related thrombus,catheter blockage,catheter pulling-off)were compared between 2 groups.Results In the experimental group,the degree of bleeding and exudation at the puncture point immediately after the operation,degree of bleeding and exudation at the puncture point 24 hours after the operation,pain score 1 day after the catheterization,pain score 3 days after the catheterization,incidence of bleeding and exudation,total incidence of complications and maintenance times were significantly lower than these in the control group(P＜0.05).In terms of the pain score immediately after the operation,pain score 5 days after the operation,incidence of phlebitis,incidence of symptomatic catheter-related thrombosis,incidence of catheter blockage,incidence of catheter pulling-off,incidence of catheter related skin injury,incidence of unplanned extubation,success rate of one-time sheath delivery and the indwelling time,the differences between the experimental group and control group were not significant(P＜0.05).Conclusion The application of blunt separation method in midline catheter indwelling can significantly reduce the incidence and degree of bleeding at the puncture point,decrease the maintenance times and relieve the pain in elderly patients with coagulation dysfunction.

Application of one-step inner-recycling counter-current chromatography for the preparative separation and purification of chemical constituents from the rhizome of Bergenia ciliate (haw.) Sternb.

Bergenia ciliata (haw.) Sternb, the renowned pharmaceutical plant in Jammu and Kashmir of Pakistan, is widely applied in treating different illnesses including diabetes, diarrhea, and vomiting. This work employed an efficient one-step inner-recycling counter-current chromatography for preparative separating and purifying compounds with similar partition coefficients from the rhizome of Bergenia ciliate (haw.). Five compounds, including quercetin rhamnodiglucoside (1), quercetin-3-O-rutinoside (2), bergenine (3), kaempferol (4), and palmatic acid (5), were successfully separated using the optimized biphasic solvent system that contained ter-butylmetylether/n-butanol/acetonitrile/water (2:2:1:5, v/v) with the purities over 98%. Mass spectrometry and nuclear magnetic resonance were conducted for structural identification. As a result, our proposed strategy might be applied in separating compounds with similar partition coefficients, which was advantageous with regard to the less solvent and time consumption, and the increased number of theoretical plates.

Isolation and purification of high polar glycosides from aerial parts of Gynostemma pentaphyllum (Thunb.) Makino by linear gradient counter-current chromatography coupled with inner-recycling mode.

Gynostemma pentaphyllum (Thunb.) Makino represents the popular health food and supplemental product with broad pharmacological activities. The highly polar glycosides, including flavonoids and saponins, are major effective active components that contain diverse sugar positions and quantities, which result in diverse chemical polarities, making it challenging to separate and isolate these components. The present work described the rapid and efficient linear gradient counter-current chromatography to preparatively separate glycosides from aboveground parts of G. pentaphyllum. Besides, the ethyl acetate and n-butanol binary mobile phases were achieved through adjusting associated proportions. Six glycosides, including quercetin-3-O-neohesperidoside (1), kaempferol-3-O-robinobioside (2), kaempferol-3-O-neohesperidoside (3), gypenoside LVI (4), ginsenoside Rb3 (5), and gypenoside XLVI (6), were isolated at the purities greater than 98%. Moreover, electrospray ionization mass spectrometry and nuclear magnetic resonance tandem mass spectrometry were conducted for structural identification. According to our findings, the established linear gradient counter-current chromatography was an efficient approach to separate the highly polar glycosides from aboveground parts of G. pentaphyllum. Our proposed strategy can be used to separate active compounds from other complex natural products.

The Preparation of Superhydrophobic Polylactic Acid Membrane with Adjustable Pore Size by Freeze Solidification Phase Separation Method for Oil-Water Separation.

An environmentally friendly pore size-controlled, superhydrophobic polylactic acid (PLA) membrane was successfully prepared by a simpler freeze solidification phase separation method (FSPS) and solution impregnation, which has application prospects in the field of oil-water separation. The pore size and structure of the membrane were adjusted by different solvent ratios and solution impregnation ratios. The PLA-FSPS membrane after solution impregnation (S-PLA-FSPS) had the characteristics of uniform pore size, superhydrophobicity and super lipophilicity, its surface roughness Ra was 338 nm, and the contact angle to water was 151°. The S-PLA-FSPS membrane was used for the oil-water separation. The membrane oil flux reached 16,084 L·m-2·h-1, and the water separation efficiency was 99.7%, which was much higher than that of other oil-water separation materials. In addition, the S-PLA-FSPS membrane could also be applied for the adsorption and removal of oil slicks and underwater heavy oil. The S-PLA-FSPS membrane has great application potential in the field of oil-water separation.

Preassigned-time projective synchronization of delayed fully quaternion-valued discontinuous neural networks with parameter uncertainties.

This paper concerns with the preassigned-time projective synchronization issue for delayed fully quaternion-valued discontinuous neural networks involving parameter uncertainties through the non-separation method. Above all, based on the existing works, a new preassigned-time stability theorem is established. Subsequently, to realize the control goals, two types of novel and simple chattering-free quaternion controllers are designed, one without the power-law term and the other with a hyperbolic-tangent function. They are different from the existing common power-law controller and exponential controller. Thirdly, under the Filippov discontinuity theories and with the aid of quaternion inequality techniques, some novel succinct sufficient criteria are obtained to ensure the addressed systems to achieve the preassigned-time synchronization by using the preassigned-time stability theory. The preassigned settling time is free from any parameter and any initial value of the system, and can be preset according to the actual task demands. Particularly, unlike the existing results, the proposed control methods can effectively avoid the chattering phenomenon, and the time delay part is removed for simplicity. Additionally, the projection coefficient is generic quaternion-valued instead of real-valued or complex-valued, and some of the previous relevant results are extended. Lastly, numerical simulations are reported to substantiate the effectiveness of the control strategies, the merits of preassigned settling time, and the correctness of the acquired results.

Separation and purification of quinolyridine alkaloids from seeds of Thermopsis lanceolata R. Br. by conventional and pH-zone-refining counter-current chromatography.

In this work, the preparative separation of quinolyridine alkaloids from seeds of T. lanceolata by conventional and pH-zone-refining counter-current chromatography. Traditional counter-current chromatography separation was performed by a flow-rate changing strategy with a solvent system of ethyl acetate-n-butanol-water (1:9:10, v/v) and 200 mg sample loading. Meanwhile, the pH-zone-refining mode was adopted for separating 2.0 g crude alkaloid extracts with the chloroform-methanol-water (4:3:3, v/v) solvent system using the stationary and mobile phases of 40 mM hydrochloric acid and 10 mM triethylamine. Finally, six compounds, including N-formylcytisine (two conformers) (1), N-acetycytisine (two conformers) (2), (-)-cytisine (3), 13-ß-hydroxylthermopsine (4), N-methylcytisine (5), and thermopsine (6) were successfully obtained in the two counter-current chromatography modes with the purities over 96.5%. Moreover, we adopted nuclear magnetic resonance and mass spectrometry for structural characterization. Based on the obtained findings, the pH-zone-refining mode was the efficient method to separate quinolyridine alkaloids relative to the traditional mode.

Constructing a Hierarchical Hydrophilic Crosslink Network on the Surface of a Polyvinylidene Fluoride Membrane for Efficient Oil/Water Emulsion Separation.

Oil/water mixtures from industrial and domestic wastewater adversely affect the environment and human beings. In this context, the development of a facile and improved separation method is crucial. Herein, dopamine was used as a bioadhesive to bind tea polyphenol (TP) onto the surface of a polyvinylidene fluoride (PVDF) membrane to form the first hydrophilic polymer network. Sodium periodate (NaIO4) is considered an oxidising agent for triggering self-polymerisation and can be used to introduce hydrophilic groups via surface manipulation to form the second hydrophilic network. In contrast to the individual polydopamine (PDA) and TP/NaIO4 composite coatings for a hydrophobic PVDF microfiltration membrane, a combination of PDA, TP, and NaIO4 has achieved the most facile treatment process for transforming the hydrophobic membrane into the hydrophilic state. The hierarchical superhydrophilic network structure with a simultaneous underwater superoleophobic membrane exhibited excellent performance in separating various oil-in-water emulsions, with a high water flux (1530 L.m-2 h-1.bar) and improved rejection (98%). The water contact angle of the modified membrane was 0° in 1 s. Moreover, the steady polyphenol coating was applied onto the surface, which endowed the membrane with an adequate antifouling and recovery capability and a robust durability against immersion in an acid, alkali, or salt solution. This facile scale-up method depends on in situ plant-inspired chemistry and has remarkable potential for practical applications.

Characterization of forehead blood flow bias on NIRS signals during neural activation with a verbal fluency task.

The major problem of near-infrared spectroscopy (NIRS) for brain activity measurement during verbal fluency task is the overlapping forehead scalp blood flow (FBF) on the target cerebral blood flow (CBF). There could be among-individual differences in the influence of FBF on CBF. We investigated effects of FBF on CBF by comparing signals obtained through a laser Doppler flowmeter (LDF) and NIRS using the modified Beer-Lambert Law (MBLL). Among 25 healthy individuals, 7 participants showed a strong correlation between LDF and NIRS signals (rs >0.500). There were no significant differences according to age or sex. Subsequently, we applied the hemodynamic separation method to the values calculated using the MBLL (Δ[oxy-Hb]M): to separate the concentration of oxygenated hemoglobin in the forehead (Δ[oxy-Hb]F) and cerebral cortex (Δ[oxy-Hb]C). First, we found that the influence of Δ[oxy-Hb]F on Δ[oxy-Hb]C in the high rs group was almost twice as large as that in the low rs group. Second, presence of sex and age differences in the influence of Δ[oxy-Hb]F on Δ[oxy-Hb]C were suggested. Based on the results, we discuss the factors affecting FBF and the resulting variations in NIRS signals.

Application of cocrystal separation technology in the separation and purification of genistein-puerarin-daidzein ternary system / 药学学报

Cocrystal separation technology is a technology that utilizes coformers to selectively form cocrystals with target compounds and separate them from mixed systems. Our study used puerarin (PUE), daidzein (DDZ), and genistein (GEN) as model drugs, which have similar structures and are the main isoflavones in Pueraria lobata root. The separation and purification processes in the modern traditional Chinese medicine (TCM) of these three components use conventional column chromatography, recrystallization, and other technologies, which have the issues of lengthy separation cycles, high solvent consumption, and inefficient preparation. Different with existing separation technology, our team used the early-found cocrystal separation method to design a step-by-step extraction and separation experiment of GEN-PUE-DDZ ternary mixture. Caffeine and L-proline were added to the mixed system in turn, GEN-caffeine cocrystal and PUE-proline cocrystal were prepared by suspension method. The cocrystals precipitated out of the solution. The purities of the GEN-caffeine cocrystal and the PUE-proline cocrystal could achieve 93% (the purity of GEN) and 99% (the purity of PUE). Besides, the purity of DDZ could also be increased by 6.76 times. This study proposed a simple operating, low cost and wide application range separation method different from the traditional separation method and realized the separation of structurally similar chemical components in TCM, laying a foundation for the application of cocrystal technology in the separation and refining of TCM.

Integration of a liquid-phase ion trap with a miniature mass spectrometer.

Liquid-phase ion trap was recently developed in our group, which has simple configuration and could realize ion manipulations in solution. In this study, a simplified liquid-phase ion trap was integrated with a home-built miniature mass spectrometer for enhanced analytical performances. Ion enrichment and separation could be realized in the liquid-phase ion trap before mass spectrometry analyses. As a result, detection sensitivity of the miniature mass spectrometer could be improved by ∼10 folds. Besides ion intensity enrichment, this liquid-phase ion trap was also capable of separating ions according to their hydrodynamic radii and effective charges. This capability was demonstrated by separating small molecules in protein background and isobaric peptide ions. Furthermore, target analytes in salt solutions and urine were also enriched and detected by the miniature mass spectrometer. Results suggest that the coupling of liquid phase ion trap with miniature mass spectrometer is a powerful technique, which could be beneficial for low abundant target molecule analysis in practical applications.

Mesoporous Ti0.5Cr0.5N for trace H2S detection with excellent long-term stability.

Efficient, accurate and reliable detection and monitoring of H2S is of significance in a wide range of areas: industrial production, medical diagnosis, environmental monitoring, and health screening. However the rapid corrosion of commercial platinum-on-carbon (Pt/C) sensing electrodes in the presence of H2S presents a fundamental challenge for fuel cell gas sensors. Herein we report a solution to the issue through the design of a sensing electrode, which is based on Pt supported on mesoporous titanium chromium nitrides (Pt/Ti0.5Cr0.5N). Its desirable characteristics are due to its high electrochemical stability and strong metal-support interactions. The Pt/Ti0.5Cr0.5N-based sensors exhibit a much smaller attenuation (1.3%) in response to H2S than Pt/C-sensor (40%), after 2 months sensing test. Furthermore, the Pt/Ti0.5Cr0.5N-based sensors exhibit negligible cross response to other interfering gases compared with hydrogen sulfide. Results of density functional theory calculation also verify the excellent long-term stability and selectivity of the gas sensor. Our work hence points to a new sensing electrode system that offers a combination of high performance and stability for fuel-cell gas sensors.

Immunological Separation of Bioactive Natural Compounds from Crude Drug Extract and Its Application for Cell-Based Studies.

In this study, we present a review on a useful approach, namely, immunoaffinity column coupled with monoclonal antibodies (MAbs), to separate natural compounds and its application for cell-based studies. The immunoaffinity column aids in separating the specific target compound from the crude extract. The column capacity was stable even after more than 10 purification cycles of use under the same conditions. After applying the crude extract to the column, the column was washed with washing buffer and eluted with elution buffer. The elution fraction contained the target compound bound to MAb, whereas the washing fraction was the crude extract, which contained all compounds except a group of target compounds; therefore, the washing fraction was referred to as a knockout (KO) crude extract. Cell-based studies using the KO extract revealed the actual effects of the natural compounds in the crude extract. One-step separation of natural compounds using the immunoaffinity column coupled with MAbs may help in determining the potential functions of natural compounds in crude extracts.

Anticorrosion Performance of PVDF Membranes Modified by Blending PTFE Nanoemulsion and Prepared through Usual Non-Solvent-Induced Phase Inversion Method.

In this study, PVDF/PTFE composite membranes were prepared by adding a PTFE nanoemulsion to a PVDF solution and casting it through the conventional non-solvent-induced phase separation method. The objective was to explore the effectiveness of using a simple and economical method to modify PVDF membranes with PTFE to enhance their anticorrosion performance, especially under highly acidic or alkaline conditions. PTFE nanoparticles (of around 200 nm in size) in nanoemulsion form were blended with PVDF at a mass ratio of PTFE:PVDF in the range of 0-0.3:1. The obtained membranes were examined to determine the effect of the added PTFE nanoparticles on the structure of the modified PVDF membranes as well as on their mechanical strength and surface characteristics. The membranes were then immersed in various concentrations of acidic or alkaline solutions for varied durations ranging from a few days up to as long as 180 days (6 months). The impacts of by the corrosive solutions on the tensile strength, surface roughness, and water flux of the membranes with different exposure times were quantified. The results showed that although a certain extent of change may occur with extended immersion times, greatly enhanced anticorrosion performance was obtained with the prepared PVDF/PTFE membranes compared with the unmodified PVDF membrane. For example, after being immersed in 5 mol-H+··L-1 H2SO4, HCl, and HNO3 solutions for 6 months, the tensile strength at breaking point remained at up to 69.70, 74.07, and 71.38%, respectively, of the initial strength for the PVDF/PTFE (M30) membrane. This was in contrast to values of only 55.77, 70.43, and 61.78% for the unmodified PVDF membrane (M0). Although the water flux and surface roughness showed a change trends to the tensile strength, the PVDF/PTFE (M30) membrane had much higher stability than the PVDF (M0) membrane. In a continuous filtration experiment containing H2SO4 at 0.01 mol-H+·L-1 for 336 h (14 days), the PVDF/PTFE membrane showed a maximum flux change of less than 30%. This was in comparison with a change of up to 50% for the PVDF membrane. However, the PVDF/PTFE membranes did not seem to have a greatly enhanced anticorrosion performance in the alkaline solution environment tested.

Chemical Adsorption Strategy for DMC-MeOH Mixture Separation.

The effective separation of dimethyl carbonate (DMC) from its methanol mixture through simple, inexpensive and low energy-input method is a promising and challenging field in the process of organic synthesis. Herein, a reversible adsorption strategy through the assistance of superbase and CO2 for DMC/methanol separation at ambient condition was described. The process was demonstrated effectively via the excellent CO2 adsorption efficiency. Notably, the protocol was also suitable to other alcohol (i.e., monohydric alcohol, dihydric alcohol, trihydric alcohol) mixtures. The study provided guidance for potential separation of DMC/alcohol mixture in the scale-up production.

Analysis of the occurrence of microplastics in beach sand on the Brazilian coast.

The occurrence of microplastics in coastal areas is a growing concern due to the increase in plastic waste pollution in recent decades. This study had as objective to evaluate the occurrence of microplastics in Brazilian beach sand, using a simple and fast methodology. In addition, possible microplastics were classified according to color, shape and chemical composition. The study was conducted on six beaches on the Brazilian coast: Pecado Beach (RJ), Castanheiras Beach (ES), Regência Beach (ES), Imbassaí Beach (BA), Viral Beach (SE) and Ponta dos Mangues Beach (SE). The methodology was adapted from international protocols, with section demarcation (100 m), where samples were collected from the superficial layer of the beach sand in ten quadrants (50 cm × 50 cm). The laboratory extraction procedure consisted of the density separation technique in two stages: the first used sodium chloride solution and the second used zinc chloride solution. For the visual inspection and screening procedures, a stereoscopic microscope was used. A total of 166 items of microplastics were collected on the six beaches on the Brazilian coast under study. The largest quantity of possible microplastics was recorded on the beaches of Viral (SE) and Ponta dos Mangues (SE) with 30.4 items/m2 and 17.4 items/m2, respectively. Regarding colour, white items were predominant (49.3%), and among particle types, fragments represented 85.1%. In addition, some microplastic particles were sent for FT-IR analysis, one of which was identified as being polyethylene. This study contributes to the knowledge of the presence of microplastics in different locations, and to supporting decision makers regarding coastal management.

Revealing Solute Clusters in Coalescence by Atom Probe Tomography Analysis.

Experimentally revealing dynamic evolution and growth behavior of small solute clusters in alloys remains a technical challenge. To date, the coalescence of the solute clusters has seldom been experimentally addressed. To address the challenge, we used atom probe tomography (APT) to access boundary information of solute clusters and identify those in close contact. By systematically investigating the population and size evolution of the clusters in close contact with aging time, we unveiled important information regarding the clusters in coalescence with the exsitu experimental technique. In this work, the maximum separation method was employed to identify clusters in APT datasets of naturally aged Al­Zn­Mg alloy. Coalescence was found to significantly contribute to the growth of small clusters and remained predominant for the formation and growth of large Guinier­Preston II ${\rm \lpar G}{\rm P}_{{\eta }^{\prime}}\rpar$ zones after 3 months aging.