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Ethylene carbonate

Ethylene carbonate is a cyclic organic compound with the chemical formula C3H4O3.
It is a clear, colorless liquid that is widely used in a variety of industrial and commercial applications, including as a solvent, electrolyte, and chemical intermediate.
Ethylene carbonate is known for its high dielectric constant, low toxicity, and biodegradability, making it a popular choice for environmentally-friendly applications.
Researchers often study ethylene carbonate to optimize its use in areas such as battery technology, polymer synthesis, and organic synthesis.
There may be typos in this description, as it is meant to appear human-written.

Most cited protocols related to «Ethylene carbonate»

1
Single-molecule FISH in Drosophila Ovaries
Cited 30 times
The following probe sets were used during smFISH: osk17×–Atto565 (DOL = 0.93), osk18×–Atto633 (DOL = 1.07), nos18×–Atto633 (DOL = 1.07), osk42×–Atto565 (DOL = 0.96), gfp23×–Atto633 (DOL = 0.94). Single-molecule FISH was performed similarly as described in Gáspár et al. (2017) (link) using ovaries of w1118 and oskar-EGFP expressing (Sarov et al. 2016 (link)) female flies. Briefly, ovaries were dissected into 2 v/v% PFA, 0.05 v/v% Triton X-100 in PBS (pH 7.4) and were fixed for 20 min on an orbital shaker. The fixative was removed and the ovaries were washed twice in PBT (PBS + 0.1 v/v% Triton X-100, pH 7.4) for 5 min. w1118 samples were treated with 2 µg/mL proteinase K in PBT for 5 min and then were subjected to 95°C in PBS + 0.05 v/v% SDS for 5 min. Specimens were cooled by adding 2× volume of room temperature PBT. Proteinase K/heat treatment was omitted in the case of oskar-EGFP expressing samples so as to preserve GFP fluorescence. Ovaries were prehybridized in 200 µL 2×HYBEC (300 mM NaCl, 30 mM sodium citrate pH 7.0, 15 v/v% ethylene carbonate, 1 mM EDTA, 50 µg/mL heparin, 100 µg/mL salmon sperm DNA, 1 v/v% Triton X-100) for 10 min at 42°C. Fifty microliters of prewarmed probe mixture (12.5–25 nM per individual oligonucleotide) was added to the prehybridization mixture, and hybridization was allowed to proceed for 2 h at 42°C. Free probe molecules were washed out of the specimen by a series of washes: 0.5 mL prewarmed 2×HYBEC, 1 mL prewarmed 2×HYBEC:PBT 1:1 mixture, 1 mL prewarmed PBT for 10 min at 42°C, and finally 1 mL prewarmed PBT allowed to cool down to room temperature. Ovaries were mounted in 80 v/v% 2,2-thiodiethanol in PBS.
Stacks of images were acquired on a Leica TCS SP8 confocal microscope using a 63× 1.4 NA oil immersion objective and were restored by deconvolution in Huygens Essential. Deconvolved images were analyzed in ImageJ using a custom-made particle detection and tracking algorithm (Gaspar et al. 2014 (link), 2017 (link)). Briefly, the algorithm finds in each slice of the reference channel the local maxima that represent the upper few percentiles of the signal distribution. These 2D objects are then connected along the z-axis based on their center-positions to create 3D objects. Signal intensities of both the reference and target channels of all 3D objects with a minimum of three slices depth (smFISH object) found within the nurse cell compartment were recorded and were subject to statistical analyses in R (as described in the legend of Fig. 3). To determine FPDR, smFISH object density in manually selected regions in the follicle cells was compared with the smFISH object density in the nurse cells in randomly selected regions of comparable volumes.
Gaspar I., Wippich F, & Ephrussi A. (2017). Enzymatic production of single-molecule FISH and RNA capture probes. RNA, 23(10), 1582-1591.
Publication 2017
2
Automated Multi-Round seqFISH+ Imaging
Cited 15 times
Imaging platform and automated fluidics delivery system were similar to those previously described with some modifications. In brief, the flow cell on the sample was first connected to the automated fluidics system. Then the region of interests(ROI) was registered using nuclei signals stained with 10 μg/mL of DAPI (D8417; Sigma). For cell culture experiments, blank images containing beads only were first imaged before the first round of serial hybridization. Each serial hybridization buffer contained three unique sequences with different concentrations of 15-nt readouts conjugated to either Alexa Fluor 647(50 nM), Cy3B(50 nM) or Alexa Fluor 488(100 nM) in EC buffer made from 10% Ethylene Carbonate (E26258; Sigma), 10% Dextran Sulfate (D4911; Sigma), 4X SSC and 1:100 dilution of SUPERase In RNase Inhibitor. The 100 μL of serial hybridization buffers for 80 rounds of seqFISH+ imaging with a repeat for round 1 (in total 81 rounds) were pipetted into a 96 well-plate. During each serial hybridization, the automated sampler will move to the well of the designated hyb buffer and flow the 100 μL hyb solution through a multichannel fluidic valves (EZ1213-820-4; IDEX Health & Science) to the flow cell (required ~25 μL) using a syringe pump (63133–01, Hamilton Company). The serial hyb solution was incubated for 17 minutes for cell culture experiments and 20 minutes for tissue slice experiments at room temperature. After serial hybridization, the sample was washed with ~300 μL of 10% formamide wash buffer (10% formamide and 0.1% Triton X-100 in 2X SSC) to remove excess readout probes and non-specific binding. Then, the sample was rinsed with ~200 μL of 4X SSC supplemented with 1:1000 dilution of SUPERase In RNase Inhibitor before stained with DAPI solution (10 μg/mL of DAPI, 4X SSC, and 1:1000 dilution of SUPERase In RNase Inhibitor) for ~15 seconds. Next, an anti-bleaching buffer solution made of 10% (w/v) glucose, 1:100 diluted catalase (Sigma C3155), 0.5 mg/mL Glucose oxidase (Sigma G2133), 0.02 U/μL SUPERase In RNase Inhibitor, 50 mM pH8 Tris-HCl in 4x SSC was flowed through the samples. Imaging was done with the microscope (Leica, DMi8) equipped with a confocal scanner unit (Yokogawa CSU-W1), a sCMOS camera (Andor Zyla 4.2 Plus), 63 × oil objective lens (Leica 1.40 NA), and a motorized stage (ASI MS2000). Lasers from CNI and filter sets from Semrock were used. Snapshots were acquired with 0.35 μm z steps for two z slices per FOV across 647-nm, 561-nm, 488-nm and 405-nm fluorescent channels. After imaging, stripping buffer made from 55% formamide and 0.1% Triton-X 100 in 2x SSC was flowed through for 1 minute, followed by an incubation time of 1 minute before rinsing with 4X SSC solution. In general, the 15-nt readouts were stripped off within seconds, and a 2-minute wash ensured the removal of any residual signal. The serial hybridization, imaging, and signal extinguishing steps were repeated for 80-rounds. Then, stainings buffer for segmentation purpose consists of 10 μg/mL of DAPI, 50nM LNA T20-Alexa 647, and 1: 100 dilution of Nissl stainings (N21480; Invitrogen) in 1x PBS was flowed in and allowed to incubate for 30 mins at room temperature before imaging. The integration of automated fluidics delivery system and imaging was controlled by a custom written script in Micro-Manager32
Eng C.H., Lawson M., Zhu Q., Dries R., Koulena N., Takei Y., Yun J., Cronin C., Karp C., Yuan G.C, & Cai L. (2019). Transcriptome-scale super-resolved imaging in tissues by RNA seqFISH+. Nature, 568(7751), 235-239.
Publication 2019
Acid Hybridizations, Nucleic alexa fluor 488 Alexa Fluor 647 Buffers Catalase Cell Culture Techniques Cell Nucleus Cells DAPI ethylene carbonate formamide Glucose Lens, Crystalline Microscopy Neoplasm Metastasis Obstetric Delivery Oxidase, Glucose Ribonucleases ribonuclease U Staining Sulfate, Dextran Syringes Technique, Dilution Tissues Triton X-100 Tromethamine
3
AAV-Mediated Neurotrophic Factor Delivery for Nerve Regeneration
Cited 10 times
AAV vector construction. Vector DNA plasmid pAAV.CMV.NT-3 (gift from B.K.K.) was used to generate single-stranded rAAV1.CMV.NT-3. It contains the human NT-3 CDS (GeneBank designation NTF3) under the control of the CMV promoter cloned between AAV2 inverted terminal repeats. To generate self-complementary (sc) AAV vectors, AAV DNA plasmid vectors pscAAV.CMV.NT-3 were generated as follows: the NT-3 coding sequence was polymerase chain reaction (PCR) amplified from plasmid, the pAAV.CMV.NT-3 vector using forward (5′-accttgcggccgccaccatgtccatcttgttttatg-3′) and reverse (5′-catatgcggccgcctcatgttcttc cgatttttctcgacaaggcacaca-3′) primers. The NT-3 PCR fragment was then digested with Not I and ligated into the self-complementary pAAV.CMV.X5 (b54) vector from which the X5 cDNA was removed by Not I digestion. For generating self-complementary DNA vector plasmid pscAAV.tMCK.NT3, the NT-3 cDNA was amplified from plasmid pAAV.CMV.NT-3 by PCR using forward (5′-atgtcggtacctgcagggatatcca ccatgtccatcttgttttatgtga-3′) and reverse (5′-tcagtggcgcgccgaaaaaacctcccacacctccc-3′) primers. The resulting NT-3 cDNA PCR fragment was then digested with Kpn I and Asc I enzymes and cloned into a self-complementary pscAAV.tMCK.aSG vector plasmid from which the αSG transgene was removed by Kpn I and Asc I digestion. The final constructs were confirmed by restriction digestion and sequencing. All vectors include a consensus Kozak sequence, an SV40 intron, and synthetic polyadenylation site (53 bp). The tMCK promoter (713 bp) was a kind gift from Dr. Xiao Xiao (University of North Carolina, Chapel Hill, NC).19 (link) It is a modification of the previously described CK6 promoter46 (link) and includes a modification in the enhancer upstream of the promoter region containing transcription factor binding sites. The enhancer is composed of 2 E-boxes (right and left). The tMCK promoter modification includes a mutation converting the left E-box to a right E-box (2R modification) and a 6 bp insertion (S5 modification).
rAAV Vector production. AAV1 vector production was accomplished using a standard 3 plasmid DNA/CaPO4 precipitation method using HEK293 cells. Two hundred and ninety-three cells were maintained in DMEM supplemented with 10% fetal bovine serum and penicillin and streptomycin. The production plasmids were: (i) pAAV.CMV.NT-3, pscAAV.CMV.NT-3, or pscAAV.tMCK.NT-3 (ii) rep2-cap1 modified AAV helper plasmid encoding the cap 1 serotype, and (iii) an adenovirus type 5 helper plasmid (pAdhelper) expressing adenovirus E2A, E4 ORF6, and VA I/II RNA genes. A quantitative PCR-based titration method was used to determine an encapsidated vg titer utilizing a Prism 7500 Taqman detector system (PE Applied Biosystems, Grand Island, NY).47 (link) The primer and fluorescent probe targeted the tMCK and CMV promoters and were as follows: tMCK forward primer, 5′-ACCCGAGATGCCTGGTTATAATT-3′; tMCK reverse primer, 5′-TCCATGGTGTACAGAGCCTAAGAC-3′; and tMCK probe, 5′-FAM-CTGCTGCCTGAGCCTGAGCGGTTAC-TAMRA-3′; CMV forward primer, 5′-TGGAAATCCCCGTGAGTCAA-3′; CMV reverse primer, 5′-CATGGTGATGCGGTTTTGG-3′; and CMV probe, 5′-FAM-CCGCTATCCACGCCCATTGATG-TAMRA-3′.
Animals, procedures and treatment groups. TrJ mice (B6.D2-Pmp22Tr-J/J) and C57BL/6 wild type were obtained from Jackson Laboratory (Bar Harbor, ME). All animal experiments were performed according to the guidelines approved by The Research Institute at Nationwide Children's Hospital Animal Care and Use Committee. The design of the experimental groups comparing single-stranded and self-complementary AAV1.NT-3 vectors, treatment duration, doses, and promoters is outlined below: (i) for the nerve regeneration study, 9–12-week-old TrJ mice were injected in the left gastrocnemius muscle with either PBS or 1 × 1011 vg of ssAAV1.CMV.NT-3 (n = 12). At 3 weeks postinjection, under isoflurane anesthesia, left sciatic nerves were exposed and crushed with a fine forceps at a level 5 mm distal to the sciatic notch to generate a regeneration paradigm as previously described.18 (link) Functional recovery, measured weekly by grip strength obtained from the limb harboring the crushed nerve and the morphological assessment of nerve regeneration were the primary endpoints of this study. At 20 weeks, postcrush mice were euthanized for tissue and serum collection for NT-3 ELISA enumeration. (ii) In this set of experiments, the effect of NT-3 gene therapy on the sciatic nerve motor conduction parameters and on the motor functions (ipsilateral and simultaneous bilateral grip strength) were investigated with endpoint correlative histopathology. Six- to 8-week-old TrJ mice received 1 × 1011 vg of ssAAV1.CMV.NT-3 or PBS in the right quadriceps muscle (n = 14 in each group). The left sciatic nerve conduction studies were performed at baseline age and were repeated at 20 and 40 weeks post-gene transfer. At 20 weeks, four vector-injected and five PBS-injected mice were euthanized for tissue collection for the assessment of NF cytoskeleton and NF phosphorylation studies using ultrastructural morphometry and western blot. Functional status of the remainder mice were monitored using rotarod between 23 and 40 weeks, and following endpoint electrophysiology, mice were euthanized for harvesting left sciatic nerve and distal leg muscles. (iii) The efficacy of scAAV1.NT-3 under control of the CMV promoter versus the muscle-specific tMCK promoter both given at three doses, within a half-log range (3 × 109 vg, 1 × 1010 vg, and 3 × 1010 vg) was assessed using endpoint electrophysiological and morphological studies. A total of 177 TrJ mice in 7 cohorts (n = 23–29 in each cohort) were generated, receiving i.m. injections of the self-complimentary vectors into the right gastric muscle at low dose, intermediate dose, or high dose with either promoters as indicated above or PBS. Technically acceptable quality nerve conduction studies were obtained from the left sciatic nerves in 171 mice. At the end of each study, mice were euthanized for tissue and serum collection for NT-3 ELISA. MF density determinations were done in high-dose cohorts (n = 13 with CMV, n = 26 with tMCK, and n = 12 with PBS).
Serum NT-3 ELISA. Serum collected from PBS and AAV1.NT-3 injected mice was assayed for NT-3 levels using a capture ELISA assay. Briefly, Immunlon4 plates were coated with 100 µl of a monoclonal anti-human NT-3 capture antibody (Cat# MAB267, 4 µg/ml, R&D Systems, Minneapolis, MN) in BupH carbonate buffer for 6 hours at 25 °C. Plates were subsequently blocked with PBS + 1% BSA + 5% sucrose overnight at 2–8 °C. The next day, plates were washed four times with PBS + 0.05% Tween20 (PBS-T) and a NT-3 standard (recombinant human NT-3, Cat# 267-N3, R&D Systems) was prepared using serial twofold dilutions in the range of 10–1,280 pg/ml in 20 mmol/l Tris, 150 mmol/l NaCl, 0.1% BSA, 0.05% Tween-20 and applied to the plate (100 µl volume). Animal sera were diluted 1:20 and 1:50 using the same dilution buffer used for the NT-3 standard and 100 µl added to the plate. Standards and serum samples were incubated at room temperature (25 °C) with gentle shaking for 2 hours ± 10 minutes. Following four PBS-T washes, 100 µl of a diluted goat anti-NT3-biotin detection antibody was added to each well and incubated 90 minutes ± 10 minutes. at RT (0.2 µg/ml of polyclonal goat anti-NT3-biotin detection antibody; Cat# BAF267; R&D Systems). Following PBS-T washes, 100 µl of a 1:1,000 dilution (PBS diluent) of extra-avidin-HRP developer solution was added to the wells and incubated for 60 minutes ± 10 minutes at RT (extra-avidin-HRP; Cat# E2896; Sigma, St Louis, MO). After washing, plates were developed by adding 100 µL of RT TMB substrate solution in the dark for 15 minutes ± 1 minutes (1-step ultra TMB-ELISA; Cat# 34028; Thermo, Waltham, MA). The reaction was stopped by adding 50 µl of 2N H2SO4, and the optical density at 450 nm determined for each well on a Bio-tek Synergy 2 ELISA plate reader running the Gen5 2.0 Data Analysis Software package (Bio-tek US, Winooski, VT). NT-3 serum concentrations were extrapolated from the NT-3 standard curve using a best fit algorithm.
Motor function testing. TrJ mice were tested for baseline motor function within 1 week prior to receiving i.m. injection of ssAAV1.CMV.NT-3 or PBS. Motor function tests included bilateral simultaneous hindlimb grip power and that of the left hind paw using a grip strength meter (Chatillon Digital Meter; Model DFIS-2; Columbus Instruments, Columbus, OH) as we have used in our previous studies.21 (link) Bilateral or unilateral grip strength was assessed by allowing the animals to grasp a platform followed by pulling the animal until it releases the platform; the force measurements were recorded in four separate trials. Measurements were performed on the same day and time of each week. Endpoint bilateral and ipsilateral grip strength measurements were done in two sessions (morning and afternoon), three trials in each per day for 3 consecutive days prior to obtaining the nerve conduction studies. The mean of these measurements were used to correlate with conduction studies.
Rotarod testing. Mouse motor function and balance was tested weekly by using the accelerating rotarod (Columbus Instruments). Mice were trained on the rotarod apparatus for 2 weeks to acclimate to testing protocol prior to data collection. A fixed rotation protocol at 5 rpm constant rotation was used, and the average of the three trials per session was recorded.
Nerve conduction studies. The nerve conduction studies were performed under isofluorane anesthesia. Temperatures were recorded with an infrared thermometer (Fisher Scientific, Pittsburgh, PA), and body temperature was maintained between 32 and 36 °C using a heating pad. Following body temperature equilibration, left sciatic nerve conduction studies were obtained using a XLTEK NeuroMaX 1002 electromyograph (Ontario, Canada) and Rhythmlink disposable subdermal needle recording electrodes (for both stimulation and recording) as we described previously.21 (link) The stimulating electrodes were placed at the proximal and distal stimulation sites (i.e., the left sciatic notch and just above the ankle, respectively), and a third pair of recording electrodes was positioned in the foot pad between the second and third digits of the left foot. The latency, duration, negative area under curve, and conduction velocity values of the recorded sciatic motor responses were determined. A caliper was utilized to measure the interelectrode distances, and these distances were used in calculations of intersegmental velocity. In addition, onset latency, duration, and amplitude were also calculated.
Processing of sciatic nerve for histopathological analysis. For the nerve regeneration study, mice were killed quickly by an overdosage of xylazine/ketamine anesthesia at 20 weeks postcrush. The sciatic nerves from crushed and intact sites were removed under a dissecting microscope, fixed in glutaraldehyde; tissue blocks were marked for proximodistal orientation and processed for plastic embedding for light and electron microscopy using standard methods established in our laboratory.48 (link) In all other experiments, left sciatic nerves were removed and processed in the same manner.
MF density determinations. Quantitative analysis at the light microscopic level was performed on 1 µm thick cross sections from regenerating and intact uncrushed sciatic nerves using a microscope-mounted video camera at ×1,600 magnification and an image analysis software (Bioquant TCW98 image analysis software; R&M Biometrics, Nashville, TN) as previously described.29 (link) Data assessing regeneration response were obtained from the second segment, at a level ~4 mm distal to the crush. The mid sciatic nerve segments were analyzed from uncrushed intact nerves in all cases. Four randomly selected areas were analyzed in each mice. MF densities (mean number ± SE/mm2) and composites of MF axon size distribution histograms were generated in rAAV1.NT-3 and PBS-injected groups.
g ratio of the MF. The g ratio refers to the ratio of axonal diameter/fiber diameter, and lower g ratios represent axons with thicker myelin.49 (link) For g ratio determinations, three representative areas of cross sectional images of mid sciatic nerves from three ssAAV1.CMV. NT-3- and PBS-injected TrJ mice and wild type were captured at ×100 magnification, and the shortest axial lengths as axon diameters and fiber diameters were recorded with a calibrated micrometer, using the AxioVision, 4.2 software (Zeiss) as we described previously.21 (link) The g ratios and axon diameters are displayed in a scattergram.
SC density. One micrometer thick, plastic embedded cross-sections were used for MF and SC nuclei counts. Three randomly selected areas in five AAV1.CMV.NT-3- and PBS-injected TrJ mice were photographed at ×100, and the number of MF and SC nuclei not in contact with the MFs was determined. Morphologic criteria used for identification of SC nuclei included homogenous, rounded, ovoid, or bean-shaped appearance with irregular contour. Nuclei with irregular contour and dense peripheral zones belonging to fibroblasts were excluded. The SC densities were estimated as number per mm2 of the endoneurial area, by adding the number of SC nuclei belonging to unmyelinated fibers or at a promyelination stage with 1:1 axon-SC relationship to the number of MFs as we reported previously.14 (link) SC nuclei belonging to the MFs were excluded.
NF packing density determinations. Ultrastructural morphometric studies were performed using cross sectional images of sciatic nerves at ×52,000 final magnification. NF density histograms were generated by determining the number of NFs per unit hexagonal area in randomly selected myelinated axons from treated and untreated TrJ mice and wild-type mice as previously described.18 (link) Ten randomly selected MFs with axon diameters between 3.6 and 5.0 µm at 20 weeks posttreatment were analyzed in each group.
Histological analysis of muscle. Gastrocnemius and tibialis anterior muscles from ssAAV1.CMV.NT-3 and PBS-injected TrJ mice (n = 3 in each group) were removed and 12 µm thick cross cryostat sections were stained for succinic dehydrogenase for generation of muscle fiber size distribution histograms as previously described.21 (link) Over 2,000 fibers were analyzed in each group.
NF cytoskeleton and phosphorylation. Sciatic and spinal nerves and roots from ssAAV1.CMV.NT-3 and PBS-injected TrJ mice were used for quantitative western blot analysis of NF proteins with NF-H-specific antibodies. Briefly, the tissues were harvested and immediately frozen over dry ice. Tissues were homogenized in radio immunoprecipitation assay buffer (50 mmol/l Tris-HCl pH 8.0, 1% NP-40, 150 mmol/l NaCl, 0.5%sodium deoxycholate, 1% sodium dodecyl sulfate, 1 mmol/l ethylene glycol tetraacetic acid, 1 mmol/l Na3VO4, 1 mmol/l NaF, phenylmethylsulfonyl fluoride (1:250), Complete protease inhibitor (1:25), and 25.5 mmol/l sodium pyrophosphate) using blue tip and Kontes pestle. Protein concentrations were determined using RC/DC method (BioRad Laboratories, Hercules, CA). For sodium dodecyl sulfate polyacrylamide gel electrophoresis, 5 µg of protein was run on 3–8% Tris-acetate NuPage gels (Invitrogen, Grand Island, NY) and transferred to PVDF membrane (Amersham Biosciences, Pittsburgh, PA). After blocking for 1 hour in 5% nonfat dry milk in TBST (100 mmol/l Tris-HCl, pH 8.0, 167 mmol/l NaCl, 0.1% Tween), the western blots were incubated with diluted primary antibodies against total NF-H (AB1989, COOH-terminal antibody from Chemicon; diluted 1:500), hyperphosphorylated NF-H (SMI-31 from Sternberger; diluted 1:20,000) and hypophosphorylated NF-H (SMI-35 from Sternberger; diluted 1:10,000). Blots were washed and incubated in appropriate horseradish peroxidase–conjugated secondary antibodies at a dilution of 1:2,000. GAPDH was used as loading control (Millipore, Billerica, MA; diluted 1:500). Immunoreactive bands were visualized with the use of ECL Plus Western blotting detection system (GE Healthcare, Pittsburgh, PA) and Hyperfilm ECL (Amersham Biosciences). Signal intensities were measured with ImageQuant software (GE Healthcare).
Statistical analysis. For comparisons between ssAAV1.CMV.NT-3 gene transfer and PBS-treated TrJ groups, statistical analysis were performed in Graph pad Prism 4 software, using one-way analysis of variance followed by Bonferroni multiple post hoc comparisons. Unpaired or paired Student's t-test was performed when applicable. Differences between the means were considered significant at two-tailed test. Significance level was set at P < 0.05. Summary statistics were reported as mean ± SEM.
For the studies comparing the efficacy of scAAV1.NT-3 under control of the CMV promoter versus the muscle-specific tMCK promoter both given at three doses, the following analyses were used: (i) Spearman correlation to study the relationship between outcomes, (ii) Kruskal–Wallis test to compare outcomes among all groups (PBS, CMV low dose/intermediate dose/high dose and tMCK low dose/intermediate dose/high dose), and (iii) Mann–Whitney U-test to compare outcomes between each group and PBS (control) group, and Bonferroni correction to adjust for multiple comparisons. Two-way analysis of variance is used to study the effects of gene vectors and doses on outcomes. All tests are conducted by SAS 9.2 (by SAS Institute, Cary, NC).
SUPPLEMENTARY MATERIALFigure S1. Serum levels of NT-3 in TrJ mice at 23 weeks postinjection (shown as individual mice) compared to PBS-treated TrJ controls (numbers 567, 570, 573, and 591) are shown in individual mice.
Figure S2. One micrometer thick, toluidine blue-stained representative cross sections of intact/uncrushed (a,b) and regenerating (c,d) sciatic nerves from TrJ mice injected with PBS (a,c) and AAV1.NT-3 (b,d) at 20 weeks. Thinly myelinated and naked axons are indicated with arrows in PBS-treated intact and regenerating nerves (a,c). AAV1.NT-3 gene therapy results in an increase of axons with thicker myelin (arrows) in intact nerves (b) and an apparent increase in the small myelinated fibers (arrows) in regenerating nerves (d).
Figure S3. Composite histograms showing myelinated fiber distribution in the regenerating (a) and contralateral intact (b) sciatic nerves from TrJ mice at 20 weeks post AAV1.NT-3 gene therapy showing an increase in the subpopulation of axons <4 µm in diameter in AAV1NT3 group compared to PBS-control.
Figure S4. Neurogenic changes in the gastrocnemius muscle from a PBS-treated TrJ (a) showing atrophic angular fibers of either histochemical fiber types (arrows) or fiber type atrophy (asterisk). Reinnervation induced changes (asterisks mark fiber type groupings) at 40 weeks post AAV1.NT-3 gene therapy (b).
Figure S5. Muscle fiber size histograms from tibialis anterior (a) and gastrocnemius (b) muscles at 40 weeks post AAV1.NT-3 gene therapy. Both muscles showed an increase in fiber diameter (c) as histologic evidence of nerve regeneration into the muscle compared to PBS-injected control group.
Figure S6. Representative tracings of sciatic motor nerve conduction from a wild-type and TrJ mouse at baseline and endpoint at 40 weeks postvector injection.
Table S1. Sciatic nerve electrophysiology in TrJ mice following AAV1. NT-3 gene transfer at 24 weeks.
Sahenk Z., Galloway G., Clark K.R., Malik V., Rodino-Klapac L.R., Kaspar B.K., Chen L., Braganza C., Montgomery C, & Mendell J.R. (2013). AAV1.NT-3 Gene Therapy for Charcot–Marie–Tooth Neuropathy. Molecular Therapy, 22(3), 511-521.
Publication 2013
4
Sequential Fluorescence In Situ Hybridization Protocol
Cited 9 times

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Publication 2018
5
Lectin-Based Fluorescence Immunoassay for Fucosylated Proteins
Cited 8 times
In our previous work, we analyzed the levels of fucosylated GP73 and fucosylated hemopexin in patients with HCC via immunoblotting of lectin-enriched fractions (19 (link)). This method involved the depletion of immunoglobulin from serum samples followed by lectin extraction of all fucosylated proteins. Subsequently, proteins were resolved through polyacrylamide gels and proteins of interest were detected via immunoblotting. As this technique was not suitable for the analysis of larger sample numbers, a lectin fluorophore-linked immunosorbent assay (FLISA) was developed.
A diagram of a typical lectin FLISA is shown in Fig. 1A. Briefly, to remove the fucosylation of the capture antibody (mouse anti-human AAT or rabbit anti-human LMW kininogen; AbD Serotec), the antibody was incubated with 10 mmol/L sodium periodate for 1 h at 4°C. An equal volume of ethylene glycol was added and the oxidized antibody was brought to a concentration of 10 μg/mL with sodium carbonate buffer (pH 9.5). Antibody (5 μg/well) was added to the plate and, following incubation, washed with 0.1% Tween 20/PBS 7.4 and blocked overnight with 3% bovine serum albumin/PBS. For analysis, 5 μL serum was diluted in 95 μL of heterophilic blocking tubes (Scantibodies Laboratory, Inc.) and incubated at room temperature for 1 h. Subsequently, samples were added to the plates for 2 h and washed five times in lectin incubation buffer [10 mmol/L Tris (pH 8.0), 0.15 mol/L NaCl, 0.1% Tween 20] before fucosylated protein was detected with a biotin-conjugated Aleuria aurantia lectin (Vector Laboratories). Bound lectin was detected using IRDye 800–conjugated streptavidin and signal intensity was measured using the Odyssey IR Imaging System (LI-COR Biotechnology). In all cases, signal intensity was compared with signals detected with commercially purchased human serum (Sigma Chemical). It is noted that the lectin FLISA detects the amount of fucosylation present on an equal amount of captured molecules from each patient sample and is done in a manner that is independent of the total amount of protein in any given patient.
Wang M., Long R.E., Comunale M.A., Junaidi O., Marrero J., Di Bisceglie A.M., Block T.M, & Mehta A.S. (2009). Novel Fucosylated Biomarkers for the Early Detection of Hepatocellular Carcinoma. Cancer epidemiology, biomarkers & prevention : a publication of the American Association for Cancer Research, cosponsored by the American Society of Preventive Oncology, 18(6), 1914-1921.
Publication 2009
Biological Assay Biotin Buffers Cloning Vectors Glycol, Ethylene Hemopexin Homo sapiens Immunoglobulins Immunosorbents IRDye800 Lectin lectin, Aleuria aurantia LMWK Mus Patients polyacrylamide gels Proteins Rabbits Serum Serum Albumin, Bovine sodium carbonate Sodium Chloride sodium metaperiodate Streptavidin Tromethamine Tween 20

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3
Electrode Materials Synthesis and Evaluation
The sodium dodecylbenzene sulfonate (SDBS, 98%) and TAPB (99.67%) were purchased from Tianjin Damao Chemical Reagent Factory, while BTMB (99.98%) and TP (97%) were obtained from Jilin Province Yanshen Technology Co, Ltd. BPY (97%) and glacial acetic acid (99.5%) were procured from Shanghai Macklin Biochemical Co, Ltd. Electrolytes (1 M LiPF6) in ethylene carbonate (EC)/dimethyl carbonate (DMC)/ethyl methyl carbonate (EMC) (volume ratio of 1:1:1) and CR2032 coin type cells were purchased from DoDochem.
Xu X., Zhang J., Zhang Z., Lu G., Cao W., Wang N., Xia Y., Feng Q, & Qiao S. (2024). All-Covalent Organic Framework Nanofilms Assembled Lithium-Ion Capacitor to Solve the Imbalanced Charge Storage Kinetics. Nano-Micro Letters, 16, 116.
Publication 2024
4
Lithium-ion Coin Cell Performance Characterization
The lithium storage performance of the as-prepared electrode materials was surveyed using CR2032 coin cells, and they were assembled in an argon-filled glove box. The working electrode was prepared using active materials comprising conductive carbon black and polyvinylidene fluoride with a mass ratio of 8:1:1, which were mixed with N-Methyl-2-pyrrolidone (NMP) and uniformly pasted on a Cu foil current collector, followed by drying in a vacuum at 120 °C for 12 h. Then, a 1 mol L−1 LiPF6 solution in a 1:1:1 mixture volume ratio comprising ethylene carbonate (EC), ethylene carbonate (DEC), and ethyl methyl carbonate (EMC) was used as the electrolyte, and lithium foil was used as the counter. The cells were charged and discharged on a Neware testing system at 0.01–3 V. CV, and EIS tests were conducted on a Parstat 4000+ electrochemical workstation (Princeton Applied Research, Princeton, NJ, USA).
Guo D., Xu Y., Xu J., Guo K., Wu N., Cao A., Liu G, & Liu X. (2024). Synergistic Engineering of CoO/MnO Heterostructures Integrated with Nitrogen-Doped Carbon Nanofibers for Lithium-Ion Batteries. Molecules, 29(10), 2228.
Publication 2024
5
Synthesis of Dibasic Esters Using Valeric Acid
A 10% by weight aqueous solution of sodium carbonate was used to wash the organic layer in the separating funnel, followed by multiple washing with distilled water. To remove xylene, under vacuum, a rotary evaporator was utilized. Then the products were dryed by using sodium sulfate anhydrous overnight12 . The codes and composition of prepared esters are given in Table 1.

The designation of prepared esters.

DesignationPrepared dibasic ester composition
A2 mol. Valeric acid + 1 mol. ethylene glycol
B2 mol. Valeric acid + 1 mol. propylene glycol
C2 mol. Valeric acid + 1 mol. butylene glycol
D2 mol. Valeric acid + 1 mol. poly ethylene glycol 400
E2 mol. Propanoic acid + 1 mol. ethylene glycol
F2 mol. Heptanoic acid + 1 mol. ethylene glycol
G2 mol. Octanoic acid + 1 mol. ethylene glycol
Shazly R.I., El-Sheshtawy H.S., Ahmed N.S, & Nassar A.M. (2024). Synthesis and biodegradation testing of some synthetic oils based on ester. Scientific Reports, 14, 3416.
Publication 2024

Top products related to «Ethylene carbonate»

1
Ethylene carbonate by Merck Group
Sourced in United States, Germany
Ethylene carbonate is a clear, colorless, and odorless organic compound. It is a cyclic carbonate ester used as a solvent and an intermediate in the production of other chemicals.
2
Sodium carbonate by Merck Group
Sourced in United States, Germany, Italy, India, France, Spain, United Kingdom, Australia, Switzerland, Poland, Portugal, China, Canada, Sao Tome and Principe, Brazil, Ireland, Mexico, Sweden, Hungary, Singapore, Malaysia, Pakistan, Thailand, Cameroon, Japan, Chile
Sodium carbonate is a water-soluble inorganic compound with the chemical formula Na2CO3. It is a white, crystalline solid that is commonly used as a pH regulator, water softener, and cleaning agent in various industrial and laboratory applications.
3
Ethylene glycol by Merck Group
Sourced in United States, Germany, United Kingdom, India, Spain, China, Sao Tome and Principe, Canada, Italy, Switzerland, France, Singapore, Poland, Sweden, Belgium, Ireland, Japan
Ethylene glycol is a colorless, odorless, and viscous liquid that is commonly used in various industrial applications. It serves as an important component in the manufacture of antifreeze, coolant, and de-icing solutions. Ethylene glycol is also utilized as a solvent and as a raw material in the production of polyester fibers and resins.
4
Sodium hydroxide (naoh) by Merck Group
Sourced in Germany, United States, India, United Kingdom, Italy, China, Spain, France, Australia, Canada, Poland, Switzerland, Singapore, Belgium, Sao Tome and Principe, Ireland, Sweden, Brazil, Israel, Mexico, Macao, Chile, Japan, Hungary, Malaysia, Denmark, Portugal, Indonesia, Netherlands, Czechia, Finland, Austria, Romania, Pakistan, Cameroon, Egypt, Greece, Bulgaria, Norway, Colombia, New Zealand, Lithuania
Sodium hydroxide is a chemical compound with the formula NaOH. It is a white, odorless, crystalline solid that is highly soluble in water and is a strong base. It is commonly used in various laboratory applications as a reagent.
5
Polyvinylidene fluoride (pvdf) by Merck Group
Sourced in United States, Germany, China, United Kingdom, India, Ireland, Morocco, Italy, Japan, Macao, France, Canada
PVDF is a type of laboratory equipment used for various applications. It is a fluoropolymer material with a unique set of properties, including chemical resistance, thermal stability, and mechanical strength. PVDF is commonly used in the manufacturing of laboratory equipment, such as filter membranes, tubing, and other components that require these specific characteristics.
6
Lipf6 by Merck Group
Sourced in United States, Germany
LiPF6 is a lithium-based compound used as a component in the electrolyte solution of lithium-ion batteries. It provides ionic conductivity and stability within the battery system.
7
Propylene carbonate by Merck Group
Sourced in Germany, United States
Propylene carbonate is a clear, colorless, and odorless liquid chemical compound. It is commonly used as a solvent in various industrial and laboratory applications.
8
Bovine serum albumin (bsa) by Merck Group
Sourced in United States, Germany, United Kingdom, China, Italy, Japan, France, Sao Tome and Principe, Canada, Macao, Spain, Switzerland, Australia, India, Israel, Belgium, Poland, Sweden, Denmark, Ireland, Hungary, Netherlands, Czechia, Brazil, Austria, Singapore, Portugal, Panama, Chile, Senegal, Morocco, Slovenia, New Zealand, Finland, Thailand, Uruguay, Argentina, Saudi Arabia, Romania, Greece, Mexico
Bovine serum albumin (BSA) is a common laboratory reagent derived from bovine blood plasma. It is a protein that serves as a stabilizer and blocking agent in various biochemical and immunological applications. BSA is widely used to maintain the activity and solubility of enzymes, proteins, and other biomolecules in experimental settings.
9
Wbcs3000 by Wonatech
The WBCS3000 is a laboratory instrument designed for the automated analysis of white blood cell (WBC) counts. It performs quantitative measurements of different types of white blood cells, providing valuable information for clinical diagnostic and research purposes.
10
N methyl 2 pyrrolidone by Merck Group
Sourced in United States, Germany, India, United Kingdom, Malaysia, Cameroon, China, Poland, Japan, Italy
N-methyl-2-pyrrolidone is a colorless, water-soluble liquid commonly used as a solvent in various industrial applications. It has a high boiling point and low volatility, making it suitable for use in a range of chemical processes.

More about "Ethylene carbonate"

Ethylene carbonate (EC) is a versatile cyclic organic compound with the chemical formula C3H4O3.
It is a clear, colorless liquid that is widely used in a variety of industrial and commercial applications, such as solvents, electrolytes, and chemical intermediates.
Ethylene carbonate is known for its high dielectric constant, low toxicity, and biodegradability, making it a popular choice for environmentally-friendly applications.
Researchers often study ethylene carbonate to optimize its use in areas like battery technology, polymer synthesis, and organic synthesis.
EC is sometimes used in combination with other compounds like sodium carbonate, ethylene glycol, sodium hydroxide, PVDF, LiPF6, propylene carbonate, and bovine serum albumin to enhance its performance in specific applications.
For example, ethylene carbonate is a key component in lithium-ion battery electrolytes, where it is often used alongside propylene carbonate and LiPF6 to improve ionic conductivity and stability.
In polymer synthesis, EC can be used as a reactive monomer or solvent to produce PVDF and other materials.
The versatility of ethylene carbonate has led to its widespread use in a variety of industries, including energy storage, electronics, and chemical manufacturing.
Researchers continue to explore new ways to utilize this unique compound, leveraging its advantageous physical and chemical properties to develop innovative solutions.