Circulating miRNAs in extracellular vesicles related to treatment response in patients with idiopathic membranous nephropathy.

BACKGROUND: Extracellular vesicle (EV)-microRNAs (miRNAs) are potential biomarkers for various renal diseases. This study attempted to identify the circulating EV-miRNA signature not only for discriminating idiopathic membranous nephropathy (IMN) from idiopathic nephrotic syndrome (INS), but also to predict the treatment response of patients with IMN. METHODS: We prospectively enrolled 60 participants, including those with IMN (n = 19) and INS (n = 21) and healthy volunteers (HVs; n = 20) in this study. Using RNA sequencing, we assessed the serum EV-miRNA profiles of all participants. To identify the EV-miRNAs predictive of treatment response in IMN, we also analyzed EV-miRNAs among patients with IMN with and without clinical remission. RESULTS: The expression levels of 3 miRNAs differed between IMN patients, INS patients and HVs. In addition, compared to HVs, RNA sequencing revealed differential expression of 77 and 44 EV-miRNAs in patients with IMN without and with remission, respectively. We also identified statistically significant (|fold change ≥ 2, p < 0.05) differences in the expression levels of 23 miRNAs in IMN without remission. Biological pathway analysis of miRNAs in IMN without remission indicated that they are likely involved in various pathways, including renal fibrosis. CONCLUSION: Our study identified EV-miRNAs associated with IMN as well as those associations with therapeutic response. Therefore, these circulating EV-miRNAs may be used as potential markers for the diagnosis and prediction of treatment response in patients with IMN.

Experimentally and spectroscopically evidenced mechanistic study of butyl peroxyacid oxidative degradation of benzo[a]pyrene in soil.

Benzo[a]pyrene (BaP) is a major indicator of soil contamination and categorized as a highly persistent, carcinogenic, and mutagenic polycyclic aromatic hydrocarbon. An advanced peroxyacid oxidation process was developed to reduce soil pollution caused by BaP originating from creosote spills from railroad sleepers. The pH, organic matter, particle size distribution of soil, and concentrations of BaP and heavy metals (Cd, Cu, Zn, Pb, and As) in the BaP-contaminated soils were estimated. A batch experiment was conducted to determine the effects of organic acid type, soil particle size, stirring speed, and reaction time on the peroxyacid oxidation of BaP in the soil samples. Additionally, the effect of the organic acid concentration on the peroxyacid degradation of BaP was investigated using an oxidizing agent in spiked soil with and without hydrogen peroxide. The results of the oxidation process indicated that BaP and heavy metal residuals were below acceptable Korean standards. A significant difference in the oxidative degradation of BaP was observed between the spiked and natural soil samples. The formation of a peroxyacid intermediate was primarily responsible for the enhanced BaP oxidation. Further, butyric acid could be reused thrice without losing the efficacy (<90%). The systematic peroxyacid oxidative degradation mechanism of BaP was also discussed. A qualitative analysis of the by-products of the BaP reaction was conducted, and their corresponding toxicities were determined for possible field applications. The findings conclude that the developed peroxyacid oxidation method has potential applications in the treatment of BaP-contaminated soils.

Design and synthesis of acrylate and acrylamide substituted pyrimidinediones as potential PPO herbicides.

PPO herbicides emerge to be widely use in the agricultural field and a focus of research to many scientists due to its environmentally-friendly properties. In lieu with this, this study presents acrylate and acrylamide substituted pyrimidinediones as PPO herbicide candidates. Most synthesized compounds exhibits herbicidal activities against both monocot and dicot weeds, especially, compound 5a which showed non-selective superior activity against the commercialized, Saflufenacil. Compound 5a was further tested for residual effect and showed promising results as shorter period is needed to cultivate the next crops. The synthesized acrylate and acrylamide substituted pyrimidinediones, especially, 5a could potentially be utilized in the development of commercial protoporphyrinogen oxidase inhibitors with further tests and studies.

Enhancement of Liposomal Plasmid DNA and siRNA Delivery by Itraconazole through Intracellular Cholesterol Accumulation.

PURPOSE: Efficient and safe vehicle that can enhance gene transfer is still needed. Since intracellular cholesterol is known to have an important role in gene delivery and itraconazole alters intracellular cholesterol trafficking, we investigated the effect of itraconazole on pDNA and siRNA delivery. METHODS: The pDNA and Bcl2 siRNA transfection efficiency was measured by luciferase assay and cytotoxicity. Cellular cholesterol was observed using filipin staining, and intracellular uptake was analyzed by flow cytometry. Lipoplex localization was observed by fluorescent labeling of DNA and lysosome after treatment of itraconazole or co-treatment of itraconazole and bafilomycin A1. RESULTS: Itraconazole enhanced the transfection efficiency of pDNA and siRNA compared to that of control through the accumulation of cholesterol. Bafilomycin A1 diminished the effect of itraconazole on gene delivery and the increment of cholesterol. Itraconazole did not increase the cellular uptake of lipoplex, but increased free pDNA during the endosome-lysosome pathway was observed during the endosome-lysosome pathway. Treating cells with both imipramine and itraconazole caused an additive effect in pDNA and siRNA delivery. CONCLUSIONS: Itraconazole enhanced gene delivery of pDNA and siRNA, and it can be used to potentiate nucleic acid therapeutics.

Dynamics of Thin-film Piezoelectric Microactuators with Large Vertical Stroke Subject to Multi-axis Coupling and Fabrication Asymmetries.

The high performance and small size of MEMS based scanners has allowed various optical imaging techniques to be realized in a small form factor. Many such devices are resonant scanners, and thus their linear and nonlinear dynamic behaviors have been studied in the past. Thin-film piezoelectric materials, in contrast, provide sufficient energy density to achieve both large static displacements and high-frequency resonance, but large deformation can in turn influence dynamic scanner behavior. This paper reports on the influence of very large stroke translation of a piezoelectric vertical actuator on its resonant behavior, which may not be otherwise explained fully by common causes of resonance shift such as beam stiffening or nonlinear forcing. To examine the change of structural compliance over the course of scanner motion, a model has been developed that includes internal forces from residual stress and the resultant additional multi-axis coupling among actuator leg structures. Like some preceding vertical scanning micro-actuators, the scanner of this work has four legs, with each leg featuring four serially connected thin-film PZT unimorphs that allow the scanner to generate larger than 400µm of vertical displacement at 14VDC. Using an excitation near one or more resonances, the input voltage can be lowered, and complementary multi-axis rotations can be also generated, but change of the resonant frequencies with scanner height needs to be understood to maximize scanner performance. The presented model well predicts the experimental observation of the decrease of the resonant frequencies of the scanner with the increase of a dc bias voltage. Also, the effects of the magnitude and uniformity of residual stress across the scanner structure on the natural frequencies have been studied.

Multi-photon vertical cross-sectional imaging with a dynamically-balanced thin-film PZT z-axis microactuator.

Use of a thin-film piezoelectric microactuator for axial scanning during multi-photon vertical cross-sectional imaging is described. The actuator uses thin-film lead-zirconate-titanate (PZT) to generate upward displacement of a central mirror platform, micro-machined from a silicon-on-insulator (SOI) wafer to dimensions compatible with endoscopic imaging instruments. Device modeling in this paper focuses on existence of frequencies near device resonance producing vertical motion with minimal off-axis tilt even in the presence of multiple vibration modes and non-uniformity in fabrication outcomes. Operation near rear resonance permits large stroke lengths at low voltages relative to other vertical microactuators. Highly uniform vertical motion of the mirror platform is a key requirement for vertical cross-sectional imaging in the remote scan architecture being used for multi-photon instrument prototyping. The stage is installed in a benchtop testbed in combination with an electrostatic mirror that performs in-plane scanning. Vertical sectional images are acquired from 15 µm diameter beads and excised mouse colon tissue.

Synthesis of cholesteryl doxorubicin and its anti-cancer activity.

Doxorubicin (dox) has been used as anti-cancer agent, but there are disadvantages such as rapid excretion, short retention time and cardiotoxicity. For giving lipophilic properties to dox, it was modified with cholesterol derivatives that were validated as a component of liposomal gene delivery. This article describes the synthesis of dox derivatives (lipo-dox A-D), their cytotoxicity and cellular uptake. In A549, HeLa, MCF7 and MDA MB 231 cell lines, lipo-dox A and lipo-dox B substituted at alcohol group showed similar anti-cancer effect as dox, but lipo-dox C and lipo-dox D substituted at amino group did not. As a result, the amino group of dox seems an important site for its cancer cell inhibition. Lipophilic property of lipo-dox A and lipo-dox B induced more accumulation in cells compared to parent drug. Therefore, the newly synthesized lipo-dox A and lipo-dox B would be a good candidate for anti-cancer agent.

DOTAP/DOPE ratio and cell type determine transfection efficiency with DOTAP-liposomes.

The effects of lipid compositions on their physicochemical properties and transfection efficiencies were investigated. Four liposome formulations with different 1,2-dioleoyl-3-trimethylammoniumpropane (DOTAP) to dioleoylphosphatidylethanolamine (DOPE) weight ratios were investigated, that is, weight ratios 1:0 (T1P0), 3:1 (T3P1), 1:1 (T1P1), and 1:3 (T1P3). Mean sizes of liposomes were influenced by their lipid composition and the preparation concentration at the time of sonication. Zeta potentials of liposomes were inversely correlated with their liposome sizes. However, neither liposome sizes nor zeta potentials were correlated with transfection efficiency. The optimum composition of liposomes was cell-line dependent (T1P0 and T3P1 for Huh7 and AGS, T3P1 and T1P1 for COS7, and T1P1 and T1P3 for A549). The shape of lipoplexes was changed from lamellar to inverted hexagonal structure according to the increased ratio of DOPE, but there was no definite advantage of specific structure in transfection efficiency throughout all used cell lines. However, cellular internalization was consistently faster in T1P0, T3P1, T1P1 compared to T1P3 in all cell lines, suggesting the importance of endosomal escape. Our findings show that the transfection efficiency of DOTAP liposomes is mainly influenced by lipid composition and cell type, and not by size or zeta potential.

Synthesis and validation of novel cholesterol-based fluorescent lipids designed to observe the cellular trafficking of cationic liposomes.

Cholesterol-based fluorescent lipids with ether linker were synthesized using NBD (Chol-E-NBD) or Rhodamine B (Chol-E-Rh), and the usefulnesses as fluorescent probes for tracing cholesterol-based liposomes were validated. The fluorescent intensities of liposomes containing these modified lipids were measured and observed under a microscope. Neither compound interfered with the expression of GFP plasmid, and live cell images were obtained without interferences. Changes in the fluorescent intensity of liposomes containing Chol-E-NBD were followed by flow cytometry for up to 24h. These fluorescent lipids could be useful probes for trafficking of cationic liposome-mediated gene delivery.

Large Stroke Vertical PZT Microactuator With High-Speed Rotational Scanning.

A thin-film piezoelectric microactuator using a novel combination of active vertical translational scanning and passive resonant rotational scanning is presented. Thin-film lead-zirconate-titanate unimorph bending beams surrounding a central platform provide nearly 200-µm displacement at 18 V with bandwidth greater than 200 Hz. Inside the platform, a mirror mount, or mirror surface, supported by silicon dioxide spring beams can be excited to resonance by low-voltage; high-frequency excitation of the outer PZT beams. Over ±5.5° mechanical resonance is obtained at 3.8 kHz and ±2 V. The combination of large translational vertical displacements and high-speed rotational scanning is intended to support real-time cross-sectional imaging in a dual axes confocal endomicroscope.

A three-degree-of-freedom thin-film PZT-actuated microactuator with large out-of-plane displacement.

A novel three degree-of-freedom microactuator based on thin-film lead-zirconate-titanate (PZT) is described with its detailed structural model. Its central rectangular-shaped mirror platform, also referred to as the stage, is actuated by four symmetric PZT bending legs such that each leg provides vertical translation for one corner of the stage. It has been developed to support real-time in vivo vertical cross-sectional imaging with a dual axes confocal endomicroscope for early cancer detection, having large displacements in three axes (z, θx, θy) and a relatively high bandwidth in the z-axis direction. Prototype microactuators closely meet the performance requirements for this application; in the out-of-plane (z-axis) direction, it has shown more than 177 µm of displacement and about 84 Hz of structural natural frequency, when two diagonal legs are actuated at 14V. With all four legs, another prototype of the same design with lighter stage mass has achieved more than 430 µm of out-of-plane displacement at 15V and about 200 Hz of bandwidth. The former design has shown approximately 6.4° and 2.9° of stage tilting about the x-axis and y-axis, respectively, at 14V. This paper also presents a modeling technique that uses experimental data to account for the effects of fabrication uncertainties in residual stress and structural dimensions. The presented model predicts the static motion of the stage within an average absolute error of 14.6 µm, which approaches the desired imaging resolution, 5 µm, and also reasonably anticipates the structural dynamic behavior of the stage. The refined model will support development of a future trajectory tracking controller for the system.

Efficient sequestration of cesium ions using phosphoric acid-modified activated carbon fibers from aqueous solutions.

In this study, acid-modified activated carbon fibers (ACF-Ps) were synthesized by phosphorylation. Three different types of ACF-based adsorbents functionalized with PO43-, P2O74-, or P3O105- ions, namely, ACF-P1, ACF-P2, and ACF-P3, were prepared by phosphorylating ACF with trisodium phosphate (Na3PO4), sodium dihydrogen pyrophosphate (Na2H2P2O5), and sodium tripolyphosphate (Na5P3O10), respectively, and utilized as adsorbents to remove cesium ions (Cs+) from aqueous solutions. Among the tested adsorbents, ACF-P3 exhibited the highest Cs+ adsorption capacity of 37.59 mg g-1 at 25 °C and pH 7 which is higher than that of ACF (5.634 mg g-1), ACF-P1 (19.38 mg g-1), and ACF-P2 (30.12 mg g-1) under the same experimental conditions. More importantly, the Cs+ removal efficiencies of ACF-P3 (82.90%), ACF-P2 (66.2%), ACF-P1 (34.2%) were 29.3-, 23.4-, and 12.11-fold higher than that of un-treated ACF (2.83%). The results suggested that the phosphorylation with Na5P3O10 is highly suitable for Cs+ adsorption which effectively functionalizes ACF with a greater number of phosphate functional groups. Adsorption and kinetic data well-fitted the Langmuir isotherm and pseudo-second-order model, respectively, which indicated the monolayer adsorption of Cs+ onto ACF-P1, ACF-P2, and ACF-P3 which were largely controlled by chemisorption. Overall, phosphoric acids containing different phosphate-based polyanions (PO43-, P2O74-, or P3O105-) enriched -OH and/or -COOH surface functional groups of ACF in addition to P-containing surface groups (PO, C-P-O, C-O-P, and P-O) and facilitated the Cs+ adsorption through surface complexation and electrostatic interactions.

Analysis of the impact of low impact development on runoff from a new district in Korea.

An analysis of the impact of a low impact development (LID) on runoff was performed using a Storm Water Management Model 5 (SWMM5)-LID model. The SWMM5 package has been developed to facilitate the analysis of the hydrologic impacts of LID facilities. Continuous simulation of urban stormwater runoff from the district which included the LID design was conducted. In order to examine the impact of runoff in the LID district the first, second and third highest ranked flood events over the past 38 years were analyzed. The assessment estimated that a LID system under historical storm conditions would reduce peak runoff by approximately 55-66% and runoff volume by approximately 25-121% in comparison with that before the LID design. The impact on runoff was also simulated under 50, 80 and 100 year return period conditions. Under these conditions, the runoff reductions within the district were estimated to be about 6-16% (peak runoff) and 33-37% (runoff volume) in comparison with conditions prior to the LID. It is concluded from these results that LID is worthy of consideration for urban flood control in future development and as part of sewer and stormwater management planning.

Stable and recyclable lanthanum hydroxide-doped graphene oxide biopolymer foam for superior aqueous arsenate removal: Insight mechanisms, batch, and column studies.

In this study, a graphene oxide-based lanthanum hydroxide/chitosan foam (CSGOL foam) was synthesized for arsenate (As(V)) remediation in surface water. The synthesized CSGOL foam texture and purity was assessed using scanning electron microscopy (SEM), Brunauer-Emmett-Teller (BET) surface area, X-ray diffraction (XRD), thermogravimetric analysis (TGA), Fourier transform infrared spectroscopy (FT-IR), and X-ray photoelectron spectroscopy (XPS) studies. The results proved that the foam was highly porous, stable, and had high surface functionality that facilitated adsorption for water pollutant removal. The sorption results proved that the As(V) removal was high (146.20 mg/g at pH 6 with 0.5 g/L CSGOL foam) when compared to the similar type of materials, endothermic chemisorption due to the production of monodentate and bidentate inner-sphere complexes. Furthermore, continuous column results indicated that the As(V) concentration in real surface waters was reduced to WHO standards (less than 10 µg As/L of water) of As(V) in drinking water for up to 10,000 bed volume. Further it can be used up to four cycles without loss of efficacy less than 93%. Because of its excellent removal capabilities and simple synthesis technique, CSGOL foam shows significant promise for treating As(V)-containing water. Further, the XPS analysis and batch studies results suggests that As(V) removal mechanism was involved electrostatic and surface complexation through chemical interaction predominately.

Development of ultra-high surface area polyaniline-based activated carbon for the removal of volatile organic compounds from industrial effluents.

Removing volatile organic compounds (VOCs) from aqueous solutions is critical for reducing VOC emissions in the environment. Activated carbons are widely used for removal of VOCs from water. However, they show less application feasibility and low removal due to less surface area. Here, a cost-effective and high surface area activated carbonized polyaniline (ACP) was synthesized to sustainable removal of VOCs from water. The ACP microstructure, surface properties, and pore structure were investigated using Brunauer-Emmett-Teller (BET) theory, Scanning Electron Microscopy (SEM), and Transmission Electron Microscopy (TEM). The specific surface area of ACP6:1 (2988.13 m2/g) was greater than that of commercial activated carbon (PAC) (1094.49 m2/g), indicating that it has excellent VOC adsorption capacity. The effects of pH, initial VOC concentration, time, temperature, and ionic strength were studied. According to kinetic and thermodynamic studies on VOCs adsorption, it is an exothermic and spontaneous process involving rate-limiting kinetics. Adsorption isotherms follow the Freundlich isotherm model, suggesting that the adsorbent surface is heterogeneous with multilayer adsorption and maximum ACP adsorption capacities of 1913.9, 2453.3, 1635.8, and 3327.0 mg/g at 293 K for benzene, toluene, ethylbenzene, and perchloroethylene, respectively, representing a 3- to 5-fold improvement over PAC. ACP is a promising adsorbent with a high adsorption efficiency for VOC removal.

Efficiency of Ppy-PA-pani and Ppy-PA composite adsorbents in Chromium(VI) removal from aqueous solution.

In this study, first time the combination of composites with Phytic acid (PA) as the organic binder cross-linker is reported. The novel use of PA with single and double conducting polymers (polypyrrole (Ppy) and polyaniline (Pani)) were tested against removal of Cr(VI) from wastewater. Characterizations (FE-SEM, EDX, FTIR, XRD, XPS) were performed to study the morphology and removal mechanism. The adsorption removal capability of Polypyrrole - Phytic Acid - Polyaniline (Ppy-PA-Pani) was deemed to be higher than Polypyrrole - Phytic Acid (Ppy-PA) due to the mere existence of Polyaniline as the extra polymer. The kinetics followed 2nd order with equilibration at 480 min, but Elovich model confirmed that chemisorption is followed. Langmuir isotherm model exhibited maximum adsorption capacity of 222.7-321.49 mg/g for Ppy-PA-Pani and 207.66-271.96 mg/g for Ppy-PA at 298K-318K with R2 values of 0.9934 and 0.9938 respectively. The adsorbents were reusable for 5 cycles of adsorption-desorption. The thermodynamic parameter, ΔH shows positive values confirmed the adsorption process was endothermic. From overall results, the removal mechanism is believed to be chemisorption through Cr(VI) reduction to Cr(III). The use of phytic acid (PA) as organic binder with combination of dual conducting polymer (Ppy-PA-Pani) was invigorating the adsorption efficiency than just single conducting polymer (Ppy-PA).

Magnetic-watermelon rinds biochar for uranium-contaminated water treatment using an electromagnetic semi-batch column with removal mechanistic investigations.

Biosorption using modified biochar has been increasingly adopted for the sustainable removal of uranium-contaminated from an aqueous solution. In this research study, the facile preparation and surface characteristics of magnetized biochar derived from waste watermelon rind to treat U(VI) contaminated water were investigated. The porosity, specific surface area, adsorption capacity, reusability, and stability were effectively improved after the magnetization of biochar. The kinetics and isotherm studies found that the U(VI) adsorption was rate-limiting monolayer sorption on the homogeneous surface of magnetized watermelon rind biochar (MWBC). The maximum adsorption capacity was found to be 323.56 mg of U(VI) per g of MWBC at pH 4.0 and 293 K that was higher than that of watermelon rind biochar (WBC) (135.86 mg g-1) and other sourced biochars. The surface interaction mechanism, environmental feasibility, applicability for real-filed water treatment studied in the electromagnetic semi-batch column, and reusability of MWBC were also explored. Furthermore, salient raised the ion exchange and complexation action capacity of MWBC due to the presence of Fe oxide. The overall results indicated that MWBC was not only inexpensive and had a high removal capacity for U(VI), but it also easily enabled phase separation from an aqueous solution, with more than three times reusability at a minimum removal capacity of 99%.

Weight Change Alters the Small RNA Profile of Urinary Extracellular Vesicles in Obesity.

INTRODUCTION: Various kidney diseases reportedly show different urinary extracellular vesicle (EV) RNA profiles. Although obesity is one of the main causes of chronic kidney disease, the expression pattern of urinary EV RNA in obesity is uncertain. Our aim was to sequence the small RNA profiles of urinary EVs in obese patients before and after weight reduction and compare them to those of healthy volunteers (HVs). METHODS: We recruited age-sex-matched obese patients and HVs. The small RNA profiles of urinary EVs were analyzed using RNA sequencing. To evaluate the effect of weight reduction, small RNA profiles of urinary EVs 6 months after bariatric surgery were also analyzed. RESULTS: The proportion of urinary EVs transfer RNA and microRNA of obese patients differed from that of HVs. Obese patients showed differential expression of 1,343 small RNAs in urinary EVs compared to HVs (fold change ≥2 and p value <0.05). Among those, 61 small RNAs were upregulated in obese patients and downregulated after weight reduction, whereas 167 small RNAs were downregulated in obese patients and upregulated after weight reduction. RNA sequencing revealed the correlation between the specific urinary EV small RNAs and clinical parameters including body weight, low-density lipoprotein cholesterol, triglyceride, high-density lipoprotein cholesterol, serum glucose, estimated glomerular filtration rate, and albuminuria. CONCLUSION: Obese patients showed distinct urinary EV small RNA profiles compared to HVs. Weight reduction altered urinary EV small-RNA profiles in obese patients.

Molecular phylogenetics of Fonsecaea strains isolated from chromoblastomycosis patients in South Korea.

Fonsecaea strains isolated from chromoblastomycosis patients in Korea and morphologically identified as Fonsecaea pedrosoi were re-evaluated for typing by sequencing the ribosomal internal transcribed spacer (ITS) regions. The ITS sequences of five Korean isolates and two reference strains were determined and then aligned with those of 11 related strains deposited in GenBank. In a phylogenetic tree constructed from these 18 strains, the Korean isolates and the references were clustered into two groups: Group A representing F. pedrosoi; Group B representing Fonsecaea monophora. These groups could be further divided into A1 and A2 subgroups and B1, B2 and B3 subgroups. Among five Korean strains, two isolates belonged to A1 subgroup, while one belonged to B1 subgroup and two to B2 subgroup. Despite the low numbers of Korean isolates and the small size of the Korean territory, this result indicates that the Fonsecaea strains prevalent in Korea are more diverse compared with those isolated in Japan and China. Moreover, F. monophora isolates, which had been reported to cause cutaneous infections as well as opportunistic neurotropic infections, were responsible for chromoblastomycosis in immunocompetent patients in Korea. In conclusion, ITS sequence analysis provided useful information not only for typing of Fonsecaea isolates in Korea but also regarding the geographical sources of these strains.