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AT 130

AT 130: A Comprehensive Approach to Advanced Research

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Most cited protocols related to «AT 130»

1
Epidemiological Surveillance in Karonga, Malawi
Cited 10 times
Karonga District is bordered to the east by Lake Malawi and to the west by the Central African (Nyika) Plateau (Figure 1). The population consists primarily of subsistence farmers, fishermen and small traders, and has grown from approximately 112,000 to over 250,000 over the past 30 years. The area was selected for its high prevalence of leprosy and its isolated situation, a factor associated with relatively little population movement. Two total population surveys were carried out in 1979–1984 and 1986–1989. 1 (link),2 (link) These surveys developed methods for precise grid mapping (later translated to GIS coordinates) and for accurate identification of individuals seen repeatedly, in different circumstances, over time. These and other methods have been continued ever since, enabling linking of interview data, clinical examinations and biological samples within and between studies. The linked databases now contain data from 800,000 contacts with 300,000 individuals.
KPS staff are stationed at all the major health facilities, where they screen inpatients and outpatients for symptoms of tuberculosis. Additionally, everyone participating in community-based studies is asked about chronic cough. Surveys have confirmed that the district-wide tuberculosis case detection rate is high. The KPS laboratory has processed all biological samples from district tuberculosis suspects since late 1985, undertaking both sputum smear microscopy and culture. Isolates have been sent to the UK for species confirmation, drug sensitivities and molecular typing. The KPS works closely with the Malawi National TB Programme and acts on its behalf in Karonga District in co-operation with the District Tuberculosis Officer, enhancing the care for tuberculosis patients in the district. The numbers of diagnosed tuberculosis cases increased greatly over the history of the KPS. The incidence of confirmed smear-positive pulmonary disease peaked in the mid-late 1990’s at around 130/100,000 adults per annum (this represents approximately 65/100,000 total population), and then declined to its current level of below 80/100,000 adults3 .
HIV data are available on the Karonga population in various contexts: clinical settings; large, ongoing case-control studies; antenatal clinic surveillance and house-to-house surveys. The earliest infections were identified from archived specimens collected in 19824 (link), and by 1988 HIV testing procedures had been established at the site. HIV prevalence was 3.9% of the 15-49 year-old population by 1988-1990, increased to 11% in 1991-1993 and reached a plateau of around 13% in 1998-20015 (link),6 .
BCG vaccination was introduced in Karonga district in mass school campaigns in the mid 1970s. Since 1990 it has been given at first health system contact, as part of the Expanded Programme on Immunization.
Crampin A.C., Glynn J.R, & Fine P.E. (2009). What has “Karonga” taught us? Tuberculosis studied over three decades. The international journal of tuberculosis and lung disease : the official journal of the International Union against Tuberculosis and Lung Disease, 13(2), 153-164.
Publication 2009
Adult AT 130 Biopharmaceuticals Central African People Cough Farmers Hypersensitivity Immunization Programs Infection Inpatient Leprosy Lung Lung Diseases Microscopy Movement Outpatients Pharmaceutical Preparations Physical Examination SERPINA3 protein, human Sputum Tuberculosis Vaccination Vision
2
Quantitative Western Blot Analysis
Cited 9 times
Samples for gel electrophoresis were reduced with 100 mM dithiothreitol in 1x Laemmli buffer (0.0625 mM Tris pH 6.8, 2% (w/v) SDS stock, 10% (v/v) glycerol, 0.002% (w/v) bromophenol blue) and heat denatured on a thermoblock at 70°C for 10 minutes. Samples were run on a 4–12% Tris-Bis NuPage Novex “mini” gel (Invitrogen) at 130 V with 1X 2-(N-morpholino)ethanesulfonic acid (MES) running buffer (Invitrogen). Gels were stained with Lumitein™ (Biotium) according to the manufacturer’s protocol.
For Western blot analysis, gels were transferred onto PVDF (Millipore) membranes, previously equilibrated in 1x NuPage transfer buffer (Invitrogen) containing 12% (v/v) methanol, at 25 V for 45 minutes and at 35 V for another 45 minutes. Membranes were blocked with 1x PBS, 0.1% Tween-20 containing 5% nonfat dry milk for 30 minutes at room temperature and probed with horseradish peroxidase conjugated mouse monoclonal anti-FLAG M2 antibody (SIGMA) (1:1000), goat C20 anti-holo and C-terminus MIS antibody (Santa Cruz) (1:200), or rabbit MGH4 anti-holo and N-terminus MIS antibody (custom) (1:1000). Blots were washed two times, 5 minutes each at room temperature with 1x PBS, Tween-20 0.1%, and incubated with appropriate secondary antibody if necessary, and then washed three times 5 minutes again. Proteins bands were visualized with the ECL kit detection system (Perkin-Elmer) onto Kodak Biomax MR film. ImageJ (NIH, http://imagej.nih.gov/ij/) was used to perform densitometry to quantify the protein bands to compare cleavage of different constructs14 (link) which was averaged over at least three independent western blots.
Pépin D., Hoang M., Nicolaou F., Hendren K., Benedict L.A., Al-Moujahed A., Sosulski A., Marmalidou A., Vavvas D, & Donahoe P.K. (2013). An albumin leader sequence coupled with a cleavage site modification enhances the yield of recombinant C-terminal Mullerian Inhibiting Substance. Technology (Elmsford, N.Y.), 1(1), 63-71.
Publication 2013
AT 130 Bromphenol Blue Buffers Cytokinesis Densitometry Dithiothreitol Electrophoresis ethane sulfonate Gels Glycerin Goat Horseradish Peroxidase Immunoglobulins Laemmli buffer Methanol Mice, House Milk, Cow's Monoclonal Antibodies Morpholinos polyvinylidene fluoride Proteins Rabbits Tissue, Membrane Tromethamine Tween 20 Western Blot
3
Design and Characterization of LNA Antisense Oligonucleotides
Cited 7 times
A series of LNA antisense oligonucleotides of different length (10- to 20-mers) were designed with 100% sequence identity to the cynomolgus monkey and human apoB mRNA sequences. The 10- to 16-mers were designed to also have 100% sequence identity to mouse apoB mRNA. (Genbank accession no.: NM_000384 and NM_009693 for human and mouse apoB mRNA respectively; the cynomolgus mRNA was sequenced in-house). All oligonucleotides were designed as gap-mers containing 8–10 DNA nucleotides flanked by 1–5 LNA nucleotides at both ends with all internucleoside linkages phosphorothioated (Table 1).

In vitro properties of oligonucleotides

Oligonucleotides
OligonucleotideaLengthTm versus RNA (°C)IC50 in Huh-7 cells (nM)
5′-TTCAGcattggtattCAGTG-3′20-mer775.0 ± 1.2
5′-CAGcattggtatTCAg-3′16-mer632.7 ± 1.3
5′-CAGcattggtatTCA-3′15-mer600.5 ± 1.3
5′-AGCattggtatTCA-3′14-mer610.2 ± 1.2
5′-GCattggtatTCA-3′13-mer570.2 ± 1.3
5′-GCattggtatTC-3′12-mer530.4 ± 1.4
5′-CattggtatT-3′10-mer44ND
5′-gcattggtattc-3′12-mer PS34ND

aGap-mer oligonucleotides with LNA (capital) and DNA (plain font). All internucleoside linkages are phosphorothioated. Melting temperatures (Tm) of LNA oligonucleotides were measured against complementary RNA. ApoB mRNA (normalized to GAPDH) IC50 values were determined from three independent experiments ( ± SD). Non-detectable IC50 values, due to low potency, marked ND.

All oligonucleotides were synthesized using standard phosphoramidite protocols on an ÄKTA Oligopilot (GE Healthcare) at 130 µmol to 8 mmol scales employing custom made polystyrene primer supports. The DNA monomers were obtained from Proligo (Sigma-Aldrich) and the LNA monomers and solid support were produced by Santaris Pharma (commercially available from Exiqon, Denmark). After synthesis, the oligonucleotides were cleaved from the support using aqueous ammonia at 65°C overnight. The oligonucleotides were purified by ion exchange and desalted using a Millipore-membrane and were finally characterized by LC-MS (Reverse phase and ESI-MS).
Straarup E.M., Fisker N., Hedtjärn M., Lindholm M.W., Rosenbohm C., Aarup V., Hansen H.F., Ørum H., Hansen J.B, & Koch T. (2010). Short locked nucleic acid antisense oligonucleotides potently reduce apolipoprotein B mRNA and serum cholesterol in mice and non-human primates. Nucleic Acids Research, 38(20), 7100-7111.
Publication 2010
Ammonia Anabolism Antisense Oligonucleotides APOB protein, human Apolipoprotein B-100 AT 130 Cells Complementary RNA GAPDH protein, human Homo sapiens Ion Exchange Macaca fascicularis Mice, House Nucleotides Oligonucleotide Primers Oligonucleotides phosphoramidite Polystyrenes RNA, Messenger Tissue, Membrane
4
Dispersing Single-Walled Carbon Nanotubes
Cited 5 times
SWCNT were dispersed by using Survanta® (Abbott Laboratories, Columbus, OH) or by the acetone-sonication method as described previously [5 (link)]). Survanta®-dispersed SWCNT (SD-SWCNT) were prepared by dispersing SWCNT (0.1 mg/ml) in PBS containing Survanta® (150 μg/ml) with light sonication (Sonic Vibra Cell Sonicator, Sonic & Material Inc, Newtown, CT, USA) at a power of 130 W, frequency of 20 kHz, and amplitude settings of 60% for 5-10 seconds. Non-dispersed SWCNT (ND-SWCNT) were prepared similarly but in the absence of Survanta®. Acetone/sonication dispersed SWCNT (AD-SWCNT) were prepared according to the method previously described [5 (link)]. Briefly, SWCNT were treated with acetone and placed in an ultrasonic bath for 24 h. The dispersed CNT were then filtered from the solution using a 20-μm nylon mesh screen followed by a 0.2-μm polytetrafluoroethylene filter. After filter collection, the dispersed CNT were washed thoroughly with distilled water and suspended in PBS with 2-3 minute sonication (Sonic Vibra Cell Sonicator, Sonic & Material Inc, Newtown, CT, USA).
Wang L., Castranova V., Mishra A., Chen B., Mercer R.R., Schwegler-Berry D, & Rojanasakul Y. (2010). Dispersion of single-walled carbon nanotubes by a natural lung surfactant for pulmonary in vitro and in vivo toxicity studies. Particle and Fibre Toxicology, 7, 31.
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Publication 2010
Acetone AT 130 Bath Cells Light Neoplasm Metastasis Nylons Polytetrafluoroethylene Survanta Ultrasonics
5
Synthesis of PEG-g-Chitosan Copolymer
Cited 5 times
The copolymer of m-PEG macromer grafted onto Cs was prepared by a modified method to that reported by Yoksan et al.32 and Yoksan and Chirachanchai.33 (link) The synthesis sequence was as follows.
Firstly, the free NH2 groups of Cs were protected via N-phthaloylation. In brief, 10 g Cs was reacted with phthalic anhydride (44.8 g, 5 mol equivalent to pyranose rings) in DMF (200 mL) at 130°C under nitrogen atmosphere for 8 h. The product of N-phthaloyl Cs (NPHCs) was then collected by filtration after precipitation in cold water, washed with methanol, and then dried under vacuum.
m-PEG was converted into m-PEG-COOH by reacting with succinic anhydride. Briefly, m-PEG (100 g, 20 mmol), succinic anhydride (2.4 g, 24 mmol), DMAP (2.44 g, 20 mmol), and triethylamine (2.02 g, 20 mmol) were dissolved in dry dioxane (350 mL). The reaction mixture was stirred at room temperature for 48 h under nitrogen atmosphere. Dioxane was evaporated under vacuum and the residue was taken up in CCl4, filtered, and precipitated by diethyl ether.
Grafting of m-PEG-COOH onto NPHCs was then performed by stirring m-PEG-COOH (37.9 g) with NPHCs (5.0 g, 0.4 mol equivalent to m-PEG-COOH) in 75 mL of DMF. HOBt (3.4 g, 3 mol equivalent to m-PEG-COOH) was then added and the stirring was continued at room temperature until obtaining a clear solution. Then, EDC·HCl (4.25 g, 3 mol equivalent to m-PEG-COOH) was added and the reaction was stirred overnight at room temperature. The reaction mixture was then dialyzed against distilled water and washed with ethanol to remove unreacted macromer.
Finally, the PEG-g-Cs copolymer was synthesized by deprotection of the NH2 groups of the PEG-g-NPHCs produced in the previous step using hydrazine monohydrate. Typically, PEG-g-NPHCs (4.0 g) was heated to 100°C with stirring under nitrogen in 15 mL of DMF. Then hydrazine monohydrate (20 mL) was added and the reaction was continued for 2 h. The mixture was then dialyzed against distilled water and ethanol and then dried under vacuum.
EL-SHERBINY I.M., MCGILL S, & SMYTH H.D. (2010). Swellable Microparticles as Carriers for Sustained Pulmonary Drug Delivery. Journal of pharmaceutical sciences, 99(5), 10.1002/jps.22003.
Publication 2010
1-hydroxybenzotriazole Anabolism AT 130 Atmosphere CCL4 protein, human Cold Temperature dioxane Ethanol Ethyl Ether Filtration hydrazine hydrate Methanol Nitrogen phthalic anhydride succinic anhydride triethylamine Vacuum

Most recents protocols related to «AT 130»

1
Synthesis and Characterization of PTMC Polymer
The PTMC polymer (molecular weight Mw ≈ 32,000 Da) was synthesized by ring-opening polymerization (Yang et al., 2015 (link)). Briefly, TMC (20.4 g, 0.2 mol) and Sn(Oct)2 (0.2 M; 1/20,000 eq, 25 μL) were added to a glass ampoule. The ampoule was then heat sealed under a high vacuum (5 mmHg) and immersed in an oil bath at 130°C ± 2°C for 24 h. The oil bath was removed, and the ampoule was cooled to room temperature. The polymer was obtained by crushing the ampoule. Subsequently, the crude polymer product was dissolved in chloroform, further purified in ice-methanol, and dried under vacuum to a constant weight. Mn and PDI were calculated using polystyrene as a standard.
Zhang J., Zhang X., Li W., Guo J., Yang L, & Yan G. (2023). Poly (trimethylene carbonate)/doxycycline hydrochloride films in the treatment of Achilles tendon defect in rats. Frontiers in Bioengineering and Biotechnology, 11, 1135248.
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Publication 2023
AT 130 Bath Chloroform Methanol poly(tetramethylene carbonate) Polymerization Polymers Polystyrenes POU2F2 protein, human Vacuum
2
Fabrication of High-Performance DNTT Transistors
To begin with, the SiO2/Si wafer was cleaned via sonication for 5 min in acetone, isopropyl alcohol, and distilled water in sequence. Subsequently, a 70 nm oxide trench was etched onto a 300 nm SiO2/Si wafer via photolithography and reactive ion etching with Ar and CF4 plasmas to form buried gate electrode patterns. While maintaining the PR for the lift-off process, a 10/60 nm Cr/Au metal layer was immediately deposited using an e-beam evaporator to fill the trench in a high vacuum chamber (~ 106 Torr). Following the deposition, the buried gate electrode was formed by lift-off of the PR using acetone, which was followed by chemical–mechanical polishing of metal residues near the edge region of the gate pattern (Fig. 1a). As a gate dielectric, 10 nm of Al2O3 layer was deposited via atomic layer deposition (ALD) at 100 °C using trimethylaluminum (TMA) and H2O precursors. To improve the quality of the Al2O3 layer, an annealing process was performed at 300 °C in high vacuum (~ 10−6 Torr) (Fig. 1b). To reduce the hysteresis of the fabricated DNTT devices, 0–20 nm thick poly (methyl methacrylate) (PMMA, Sigma-Aldrich, Mw = 350,000) buffer layers were applied as a primer layer before the deposition of DNTT. The PMMA layer prevents the charging effect between the inorganic dielectric and DNTT [31 (link)]. To prepare the PMMA solution, PMMA powder was dissolved in toluene solvent, and the solution was stirred at 70 °C overnight to completely dissolve the PMMA powder. Thereafter, the PMMA solution was coated on top of the Al2O3 layer by spin-coating at 3000 rpm for 60 s. Subsequently, the solution was baked at 70 °C for 10 min to remove the remaining solvent (Fig. 1c). The thickness of the PMMA layer was modulated by varying the concentration of the PMMA solution. The thickness of the PMMA layer was measured using an ellipsometer and via atomic force microscopy (AFM). Following the PMMA coating, a 40 nm thick DNTT layer (Sigma-Aldrich) was thermally deposited in high vacuum, followed by baking at 130 °C for 20 min in ambient air (Fig. 1d). The PMMA layer became more resistant to PR removal during the high-temperature baking process (Additional file 1: Fig. S1).

a–h Schematic of the fabrication process of the DNTT TFT based on the photolithography process. Photographs of the DNTT TFT fabricated via (i) photolithography and (j) shadow mask method, where W/L = 12 μm /6 μm and 200 μm /100 μm, respectively. Scale bars are 20 μm (i) and 200 μm (f). (k) ID–VG curves of DNTT TFTs developed using different fabrication methods for VD =  − 2 V. (l) Comparison of currents, field-effect mobility, and threshold voltage of the fabricated devices

For the DNTT/separation layer (SL) device and DNTT ternary logic device, a 1.5 nm Al2O3 layer was deposited onto the DNTT layer via ALD, followed by 0–20 nm of PMMA coating. Subsequently, the second 40 nm DNTT channel layer was additionally deposited for the DNTT ternary logic device.
For all the devices, the organic stack structures (DNTT, DNTT/SL, and DNTT ternary logic device) were simultaneously patterned. For channel patterning, a 30 nm Au hardmask was deposited via a thermal evaporation process. Contact photolithography was performed with a minimum critical dimension of ~ 2 µm (Fig. 1e). The exposed Au regions were etched using a gold etchant TFA (Transene) at 25 °C for 10 s. Thereafter, the PR mask was completely removed using dimethyl sulfoxide (DMSO) (Fig. 1f). Subsequently, the DNTT channel pattern was etched with an Au hardmask pattern via an oxygen plasma etching process (RF power = 50 W, O2 pressure = 350 mTorr). Here, the Au hardmask blocks UV light during photolithography and plasma processes [32 (link)], preventing degradation caused by the positive threshold voltage (Vth) shift [33 (link)]. Subsequently, 70 nm of Au was blanket-deposited. Finally, another photolithography technique was applied to form source/drain (S/D) electrode patterns (Fig. 1g). Using the PR mask, S/D electrode patterns were formed via the Au wet etching process for 30 s, followed by PR mask removal (Fig. 1h). At the end of each step, the results of the process were examined using an optical microscope to ensure the robustness of the pattern (Additional file 1: Fig. S2).
As a reference, another DNTT TFT device with shadow masks that did not go through any patterning or etching processes of the DNTT channel and S/D electrodes was fabricated. The buried gate electrode, Al2O3 dielectric, and PMMA buffer layer were formed on SiO2/Si substrate, which is exactly the same as the process described above. Then the DNTT channel layer was deposited using a shadow mask and baked at 130 °C for 20 min. Lastly, 100 nm of Au was deposited using another shadow mask to form S/D electrodes. The W and L of the device fabricated via the lithography process were 12 and 6 μm, respectively (Fig. 1i), whereas those of the devices fabricated using shadow masks were 200 and 100 μm, respectively (Fig. 1j).
The final devices were electrically characterized using a semiconductor parameter analyzer (Keithley 4200) at room temperature under ambient air conditions.
Lee Y., Kwon H., Kim S.M., Lee H.I., Kim K., Lee H.W., Kim S.Y., Hwang H.J, & Lee B.H. (2023). Demonstration of p-type stack-channel ternary logic device using scalable DNTT patterning process. Nano Convergence, 10, 12.
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Publication 2023
Acetone Air Conditioning AT 130 Buffers Electricity Fever Gold Isopropyl Alcohol Light Microscopy Medical Devices Metals Microscopy, Atomic Force Oligonucleotide Primers Oxides Plasma Polymethyl Methacrylate Powder Pressure Range of Motion, Articular Solvents Sulfoxide, Dimethyl Toluene Ultraviolet Rays Vacuum
3
Generation and Characterization of Labeled DNA Probes
For generation of the labeled probe, 5ʹ-IRDye® 700 labeled oligonucleotides were purchased from IDT with the following sequences: ABE; 5ʹ-CGG TGT TGC ACG CGG *CGG GAC GCT CGC GGT AGT TTT* TTC CCA TGA TCA CG-3ʹ and 5ʹ-CGT GAT CAT GGG AAA *AAA CTA CCG CGA GCG TCC CGC CGC* GTG CAA CAC CG-3ʹ and scrambled control probes; 5ʹGTT TAC TAG GTC GAG GTA CTT CGA CGC GCG CCG TCT GCT AGC GCG GTC TG-3ʹ and 5ʹ-CA GAC CGC GCT AGC AGA CGG CGC GCG TCG AAG TAC CTC GAC CTA GTA AAC3ʹ. The AlpA binding element is indicated by asterisks. The oligonucleotides were annealed by mixing them in equimolar amounts in duplexing buffer (100 mM Potassium Acetate; 30 mM HEPES, pH 7.5) and heating to 100 °C for 5 min in a PCR cycler. The cycler was then turned off and the samples were allowed to cool to room temperature while still inside the block. The annealed product was then diluted with water to 6.25 nM for EMSA experiments.
For EMSAs fluorophore labeled DNA probes at a concentration of 0.3125 nM were incubated for 30 min at 20 °C in 20 µl reaction mix (Licor Odysee EMSA Kit) containing 33.4 mM Tris, 70.2 mM NaCl, 12.5 mM NaOAc, 3.75 mM HEPES, 50 mM KCl, 3.5 mM DTT, 0.25% Tween20 and 0.5 µg sheared salmon sperm DNA (ThermoFisher) with proteins. For resolving the reactions, 4% polyacrylamide gels containing 30% triethylene glycol were cast (For two gels: 2 ml ROTIPHORESE®Gel 30 37.5:1 (Roth), 4.5 ml triethylene glycol (Sigma-Aldrich), 1.5 ml 5x TBE-buffer, 7 ml ddH2O, 15 µl TEMED, 75 µl 10% APS). The gels were preequilibrated for 30 min at 130 V in 0.5x TBE-buffer. Samples with added 10x orange dye were then loaded onto the gel at 4 °C and the voltage set to 300 V until the samples entered the gel completely. The voltage was then turned down to 130 V and the gel was run until the migration front reached the end of the gel. The gels were imaged using the Licor Odyssey imaging system using the 700 nm channel. For generation of the figures, the scanned image was converted to greyscale and brightness and contrast adjusted. The unprocessed scan is available as Supplementary Fig. 15.
Eggers O., Renschler F.A., Michalek L.A., Wackler N., Walter E., Smollich F., Klein K., Sonnabend M.S., Egle V., Angelov A., Engesser C., Borisova M., Mayer C., Schütz M, & Bohn E. (2023). YgfB increases β-lactam resistance in Pseudomonas aeruginosa by counteracting AlpA-mediated ampDh3 expression. Communications Biology, 6, 254.
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Publication 2023
5'-chloroacetamido-5'-deoxythymidine AT 130 Buffers CD3EAP protein, human Electrophoretic Mobility Shift Assay Gels HEPES Oligonucleotides polyacrylamide gels Potassium Acetate Proteins Radionuclide Imaging Salmo salar Sodium Chloride Sperm triethylene glycol Tris-borate-EDTA buffer Tromethamine Tween 20
4
Monitoring Deltamethrin Dispersal in Aquatic Ecosystems
The delousing at site 1 took place in the pens
during 3 days in early spring in 2020, and a total of 27 L of Alpha
Max was used, which corresponds to 270 g of deltamethrin for the full
treatment. The site had not been treated with deltamethrin since 2017.35 PAS made of AlteSil silicone rubber sheets
(thickness: 0.5 mm, purchased from Altec, UK) were spiked with performance
reference compounds (PRCs; details in the SI) prior to deployment. These non-naturally occurring compounds are
used to estimate in situ contaminant exchange kinetics
between water and silicone rubber. The samplers (A–E) were
deployed 3 days before delousing, 15–120 m from the deloused
pens at 3–5 m and 10–14 m depth at five sampling sites
(n = 10). PAS were collected 3 days after the delousing
ended (Table S2). Sampler A, situated inside
a deloused pen, was lost, but samplers B–E were recovered and
stored frozen in tin containers until analyses. Surface sediment samples
(n = 10) were collected 5 weeks after the last delousing
event at distances 0–500 m from the pens at 70–130 m
depth with a Van Veen grab, and the 0–1 cm top layer was transferred
to preburned (450 °C) glasses and kept frozen until analyses
(Table S2). Only grabs with an undisturbed
surface were approved for sampling.
Arnberg M., Refseth G.H., Allan I.J., Benedetti M., Regoli F., Tassara L., Sagerup K., Drivdal M., Nøst O.A., Evenset A, & Carlsson P. (2023). Acute and Sublethal Effects of Deltamethrin Discharges from the Aquaculture Industry on Northern Shrimp (Pandalus borealis Krøyer, 1838): Dispersal Modeling and Field Investigations. Environmental Science & Technology, 57(9), 3602-3611.
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Publication 2023
AT 130 decamethrin Eyeglasses Freezing Silicone Elastomers
5
Cryo-EM Structural Analysis of SEQ-9-bound Mtb 70S Ribosomes
SEQ-9-bound A2296-methylated Mtb 70S was prepared by incubating 800 nM methylated 70S with 16 μM SEQ-9 on ice for 30 min. 3 μl of the sample was applied to a glow-discharged Quantifoil R2/2 holey carbon grid (300 mesh), and vitrified using a Vitrobot Mark III (FEI company, The Netherlands) at 22°C with 100% relative humidity. Cryo-EM data were collected on a Titan Krios electron microscope (Thermo Fisher, USA) operating at 300 kV, at a nominal magnification of 130,000, which yields a pixel size of 1.06 Å/pixel. Image stacks were recorded on a Gatan K2 Summit (Gatan, Pleasanton CA, USA) direct detection camera in the electron counting mode. A total exposure time of 8.0 s with 0.2 s intervals and dose rate of ∼6.0 electrons/ Å2/s were used, resulting in 40 frames per image stack and accumulated total dose of ∼48 electrons per Å2.
Drift correction of collected image stacks was done by MotionCor2 with dose weighting.44 (link) Aligned and summed image stacks were subjected to CTF estimation using gCTF.45 (link) Images showing ice contamination and low resolution according to the estimation from gCTF were discarded, resulting in a total of 6,481 selected micrographs. Particle picking and reference-free 2D class average were done by Gautomatch (http://www.mrc-lmb.cam.ac.uk/kzhang/) and RELION-3.0,49 (link) respectively. A total of 1,065,544 clean particles were selected and refined into one consensus map. Then 3D classification was used with the option ‘--skip_align’ to classify different states of ribosomes, and each state was processed according to the pipeline of RELION-3.0. The overall resolution was estimated according to the gold-standard Fourier shell correlation.50 (link)
SEQ-9-bound unmethylated Mtb 70S was prepared by incubating 700 nM unmethylated 70S with 16 μM SEQ-9 on ice for 30 min. 3 μl of the sample was applied to a glow-discharged Quantifoil R2/1 holey carbon grid (300 mesh) with 2nm carbon film, and vitrified using a Vitrobot Mark III (FEI company, The Netherlands) at 22°C with 100% relative humidity. Cryo-EM data were collected under a Titan Krios electron microscope (Thermo Fisher, USA) operating at 300 kV, at a nominal magnification of 165,000, which yields a pixel size of 0.83 Å/pixel. Image stacks were recorded on a Gatan K2 Summit (Gatan, Pleasanton CA, USA) direct detection camera in the electron counting mode. A total exposure time of 7.0 s with 0.2 s intervals and dose rate of ∼6.0 electrons/ Å2/s was used, resulting in 35 frames per image stack and accumulated total dose of ∼42 electrons per Å2.
Drift correction of collected image stacks was done by MotionCor2 with dose weighting.44 (link) Aligned and summed image stacks were subjected to CTF estimation using Cryosparc Patch CTF estimation function.39 (link) Images showing ice contamination and low resolution according to the estimation from Patch CTF were discarded, resulting in a total of 11,621 selected micrographs. Particle picking and reference-free 2D class average were done by Cryosparc.39 (link) A total of 318,826 Mtb 70S particles were selected and refined into one consensus unmethylated Mtb 70S ribosome map at 2.8 Å (Figure S3A). To improve the resolution of the drug-binding site, we combined another 400,424 Mtb 50S particles in the same dataset from the same purification, yielding a total number of 719,250 particles of 70S and 50S. The drug-binding site was far away from the ribosomal subunit interface and was conserved in both the 70S and 50S, justifying the strategy to combine the 70S and 50S data to improve the resolution of the drug-binding site (Figure S3B). By applying a mask around the 50S subunit of the data mixing 70S and 50S, we obtained an unmethylated Mtb 50S ribosome map at 2.6 Å. The overall resolutions were estimated according to the gold-standard Fourier shell correlation (Relion 0.143 criterion).50 (link) Final Fourier Shell Correlation plots for the density maps were generated using MATLAB R2021a.
Zhang J., Lair C., Roubert C., Amaning K., Barrio M.B., Benedetti Y., Cui Z., Xing Z., Li X., Franzblau S.G., Baurin N., Bordon-Pallier F., Cantalloube C., Sans S., Silve S., Blanc I., Fraisse L., Rak A., Jenner L.B., Yusupova G., Yusupov M., Zhang J., Kaneko T., Yang T.J., Fotouhi N., Nuermberger E., Tyagi S., Betoudji F., Upton A., Sacchettini J.C, & Lagrange S. (2023). Discovery of natural-product-derived sequanamycins as potent oral anti-tuberculosis agents. Cell, 186(5), 1013-1025.e24.
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Publication 2023
AT 130 Binding Sites Carbon Electron Microscopy Electrons Gold Gold-50 Humidity MAP2 protein, human Microtubule-Associated Proteins Pharmaceutical Preparations Protein Subunits Reading Frames Ribosomes Ribosome Subunits

Top products related to «AT 130»

1
Vitrobot mark 4 by Thermo Fisher Scientific
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Epu software by Thermo Fisher Scientific
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EPU software is a platform developed by Thermo Fisher Scientific for the acquisition and processing of cryo-electron microscopy (cryo-EM) data. The software provides a user-friendly interface to control the microscope and automate the image acquisition process, ensuring consistent and reliable data collection.
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Titan krios microscope by Thermo Fisher Scientific
Sourced in United States, United Kingdom, Germany
The Titan Krios microscope is a high-performance cryo-electron microscope (cryo-EM) designed for advanced structural biology research. It features a stable, high-resolution electron beam and sophisticated imaging capabilities to enable the visualization and analysis of biological samples at the atomic level.
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K2 summit by Ametek
Sourced in United States, France, United Kingdom
The K2 Summit is a high-performance laboratory equipment designed for precise analysis and measurement. It features advanced capabilities for accurate data collection and analysis, enabling researchers and scientists to conduct their work efficiently and effectively.
5
Titan krios electron microscope by Thermo Fisher Scientific
Sourced in United States, United Kingdom
The Titan Krios electron microscope is a high-resolution transmission electron microscope designed for advanced structural biology research. It features a stable and powerful electron beam, advanced optics, and a state-of-the-art detector system, enabling the visualization and analysis of biological samples at the atomic level.
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Talos arctica by Thermo Fisher Scientific
Sourced in United States
The Talos Arctica is a high-performance cryo-transmission electron microscope (cryo-TEM) designed for structural biology research. It features a stable and efficient cryogenic system, advanced optics, and a powerful digital camera for obtaining high-resolution images of biological samples.
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Titan krios cryo electron microscope by Thermo Fisher Scientific
Sourced in United States
The Titan Krios cryo-electron microscope is a high-performance instrument designed for advanced structural biology applications. It is capable of producing high-resolution images of biological samples by using a combination of cryogenic temperatures and advanced electron optics.
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Titan krios by Thermo Fisher Scientific
Sourced in United States, United Kingdom, Netherlands, Germany, China, Sweden
The Titan Krios is a high-performance transmission electron microscope (TEM) designed for cryo-electron microscopy (cryo-EM) applications. It provides high-resolution imaging and data acquisition capabilities for the study of biological macromolecular structures.
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Epu 2 by Thermo Fisher Scientific
Sourced in United States
EPU 2 software is a data collection and processing platform for transmission electron microscopy (TEM) instruments. It provides an intuitive user interface for automated data acquisition and processing workflows. The software's core function is to facilitate the efficient collection and management of high-quality TEM images and data.
10
Bioquantum energy filter by Ametek
Sourced in United States
The BioQuantum energy filter is a laboratory equipment designed to filter and condition energy inputs for a variety of applications. It functions by regulating and stabilizing the energy supply to ensure consistent and reliable performance of connected devices.

More about "AT 130"

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AT 130 is a comprehensive approach to advanced research, covering a wide range of topics and techniques.
This includes the use of specialized equipment like the Vitrobot Mark IV for sample preparation, EPU software for data acquisition, and the Titan Krios microscope with a K2 Summit direct electron detector for high-resolution cryo-electron microscopy (cryo-EM).
The Titan Krios electron microscope, Talos Arctica, and other Titan Krios cryo-electron microscopes are also commonly used in AT 130 studies.
The EPU 2 software and the BioQuantum energy filter are additional tools that can enhance the quality and efficiency of AT 130 research.
By leveraging these advanced technologies and techniques, researchers can gain deeper insights into the structural and functional characteristics of their samples, leading to groundbreaking discoveries and innovations in their respective fields.
Whether you're a seasoned researcher or just starting your AT 130 journey, PubCompare.ai can help you navigate the latest advancements and unlock new possibilities in your studies.
Explore our intelligent comparison tools and discover the optimal protocols to elevate your AT 130 research to new heights.