Notes

Recombinant γY278H Fibrinogen Revealed Typical Release coming from CHO Tissues, but a Related Heterozygous Individual Demonstrated Hypofibrinogenemia.
Effective process, including a cartridge packing polypropylene fiber sorbent modified by following on-line polydopamine coating, for on-line solid phase extraction in 2D UHPLC system has been developed. Hydrophobic surface of mechanically stable polypropylene fibers was hydrophilized using an automated and reproducible in situ coating process to enable good wettability and effective extraction of polar compounds. Polymerization mixture consisting dopamine and TRIS buffer was circulated through the cartridge containing polypropylene fibers using a peristaltic pump to achieve polymerization. This process was optimized in terms of dopamine amount in the polymerization mixture, its flow rate, and polymerization time. Best results were obtained with 25 mL polymerization mixture containing 20 mg dopamine circulated through the cartridge at a flow rate of 2.07 mL min-1 for 60 min. Prepared cartridges were evaluated via measurement of the recovery and reproducibility using chlorogenic acid as a model compound. Overall reproducibility of our multistep process including eight cartridges in 2D UHPLC system, each measured in triplicate, was 3.61% (n = 24).We developed a biochemical gas sensor (bio-sniffer) using the enzymatic reaction of alcohol dehydrogenase (ADH) to target ethanol in skin gas. By introducing a gas concentrator using liquid nitrogen, we constructed a highly sensitive system for skin gas measurements. Eganelisib The ethanol bio-sniffer was built from an optical-fiber probe employing an ADH enzyme membrane, an UV-LED light source for excitation, and a photomultiplier tube. Ethanol was measured by detecting the autofluorescence of the coenzyme NADH due to the enzymatic reaction of ADH. We established a system for measuring concentrated gases by connecting the sensor with a gas concentrator and introducing concentrated skin gas to the sensing surface. This suppressed diffusion of the concentrated gases to achieve maximum fluorescence intensity by optimizing the measurement system. The calibration curve from obtained peak values showed ethanol gas can be measured over 1-3100 ppb, which included skin gas concentrations during alcohol consumption. Finally, when applied to measurements of ethanol in skin gas following alcohol consumption, the output was found to be dependent on concentration, similarly to using standard gases. Consecutive measurements were possible using periodic sampling with 6-min intervals for 180 min of monitoring. Skin ethanol concentrations rose from 20 min after consuming the alcohol, exhibited a peak value of 25 ppb skin gas ethanol at around 60 min, and gradually declined. Thus, the system can be used for non-invasive percutaneous evaluation of human volatile organic chemicals in blood.Comprehensive two-dimensional gas chromatography coupled with time-of-flight mass spectrometry (GC × GC/TOFMS) is used to characterize complex bio-oil samples because of the high peak capacity associated with the high acquisition rate and mass spectra deconvolution capability of TOFMS. A recent application of fast GC × GC for this type of analysis improved sample throughput while achieving the same peak capacity without the use of cryogenic liquids. This work evaluates the effect of the TOFMS data acquisition rate on the quality of the analytical information obtained by GC × GC/TOFMS. In the analysis of coconut fiber bio-oil under fast GC × GC/TOFMS conditions, use of high data acquisition rates (200-300 Hz) increases the number of identifiable peaks by more than 50% compared with that achieved at the conventional rate of 100 Hz. The acquisition rate can affect the peak capacity by a factor of 3 or more. This is the first study to demonstrate the importance of optimizing the data acquisition rate, a parameter that has previously been neglected in the literature, in GC × GC/TOFMS development.An original, selective and automated method, for the microextraction by packed sorbent (MEPS) of Δ9-tetrahydrocannabinol (THC), 11-hydroxy-Δ9-tetrahydrocannabinol (THC-OH), and 11-nor-Δ9-tetrahydrocannabinol-9-carboxylic acid (THC-COOH) from human urine, was developed by using (i) a catechin-molded molecularly imprinted polymer (MIP), (ii) a new lab-made MEPS device easily repackable with any commercial or lab-made sorbent, and (iii) a lab-made multi syringe autosampler. Analyses were performed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and the developed method proved to be precise, accurate and showed good linearity. Determination coefficients ranged from 0.96 to 0.99, in the range of 5-250 ng mL-1. Limits of detection and quantification ranged between 1.0 and 5.0 ng mL-1 and 5.0 and 20.0 ng mL-1. The method was successfully applied in the analysis of real urine samples. The same packed syringe was effectively used over 90 consecutive extractions without carry-over effects.Matrix-assisted laser desorption ionization (MALDI) imaging mass spectrometry (IMS) is increasingly recognized for its potential in the discovery of novel biomarkers directly from tissue sections. However, there are no MALDI IMS studies as yet on the adipose tissue, a lipid-enriched tissue that plays a pivotal role in the development of obesity-associated disorders. Herein, we aimed at developing an optimized method for analyzing adipose tissue lipid composition under both physiological and pathological conditions by MALDI IMS. Our studies showed an exacerbated lipid delocalization from adipose tissue sections when conventional strategies were applied. However, our optimized method using conductive-tape sampling and 2,5-dihydroxybenzoic acid (DHB) as a matrix, preserved the anatomical organization and minimized lipid diffusion from sample sections. This method enabled the identification of a total of 625 down-regulated and 328 up-regulated m/z values in the adipose tissue from a rat model of extreme obesity as compared to lean animals. Combination of MALDI IMS and liquid chromatography (LC)-MS/MS data identified 44 differentially expressed lipid species between lean and obese animals, including phospholipids and sphingomyelins. Among the lipids identified, SM(d180_182), PE(P-160_200), and PC(O-160_161) showed a differential spatial distribution in the adipose tissue of lean vs. obese animals. In sum, our method provides a valuable new tool for research on adipose tissue that may pave the way for the identification of novel biomarkers of obesity and metabolic disease.
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