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In this work, we used a titanium-based metal-organic framework (MOF, Ti-MIL125-NH2) as a novel enrichment platform to detect protein kinase A (PKA) activity and to screen relevant kinase inhibitors. This method took advantage of the highly specific recognition of phosphate groups by the Ti-MIL125-NH2 nanoparticle. In the presence of PKA and adenosine 5'-triphosphate (ATP), the fluorophore-labeled peptide substrate was phosphorylated, and the generated phosphopeptide could then specifically bind to the titanium sites of Ti-MIL125-NH2. This resulted in fluorescence enrichment, which could be efficiently detected by the system. Under optimal conditions, the method presented a linear relationship in the experimental range of 0.00005-0.01 U μL-1, and the limit of detection was 0.00003 U μL-1 (3σ, n = 11). Furthermore, protein kinase Akt1 was tested to verify the universality of this method. The method was also successfully applied in cell lysates for kinase activity analysis and inhibitor screening. Thus, the new, highly sensitive fluorescence method based on MOF for detecting PKA activity is an excellent tool that has potential applications in kinase-related disease and basic research.Herein, a kind of novel hemin-based metal organic frameworks (Fe-hemin-MOFs) with unique peroxidase-like bioactivity was developed for the first time. Orforglipron purchase The synthesized Fe-hemin-MOFs exhibited satisfactory catalytic activity toward hydrogen peroxide (H2O2). When it was further supported on Chitosan-reduced graphene oxide (CS-rGO), amplified electrochemical signal could be obtained. The Fe-hemin-MOFs/CS-rGO composite was used to construct a novel H2O2 electrochemical sensor. The electrocatalytic reduction of H2O2 displayed two segments linearity range from 1 to 61 μM and 61-1311 μM, as well as a low detection limit of 0.57 μM. Furthermore, the proposed sensor was successfully used for real-time monitoring of H2O2 released from living cells, which extended the practical application of MOFs-based sensors in monitoring the pathological process in living cells.Herein, we report a rational design in preparation of natural indicator incorporated media used as a rapid, selective, sensitive and economic colorimetric biosensor for detection of Methicillin-resistant Staphylococcus aureus (MRSA). Anthocyanins obtained from red cabbage (Brassica oleracea) extract (RCE) are major components and acted as pH responsive indicators in the RCE media. The RCE media was tested for eight gram-positive, four gram-negative bacterial and one model fungal pathogens. We experimentally revealed how salt concentration and antibiotic (cefoxitin) in the RCE media influence the growth of pathogens and especially MRSA. Benefiting pH dependent color change function of anthocyanins is indispensable point of the RCE media in sensing of MRSA growth. The potential MRSA colorimetric sensing mechanism of anthocyanins relies on both protonation and decrease in electron density on structures of anthocyanins by acidic organic volatile compounds produced during growth of MRSA. The protonated anthocyanins with low electron density changed original purple color of the RCE media to pink color. We demonstrated that detection of MRSA was achieved by spectrophotometrically (one single cell) and visually with naked eyes (100 cells) in 2nd hrs and 6th hrs of incubation, respectively. We optimized concentrations of cefoxitin (from 1 μg/mL to 20 μg/mL) against MRSA and Methicillin-sensitive Staphylococcus aureus (MSSA) bacterial cell (3 McFarland) suspensions, then growth of MRSA was visually and clearly detected in the presence of 4 μg/mL cefoxitin between 90 min and 3 h. We persistently offer that the proposed RCE media can be a well alternative to currently used phenotypic methods in clinics owing to its much rapid, sensitive, selective and economic properties.Single atom nanozymes are the artificial enzymes with enzyme-like activity, which have attracted a great deal attention in recent years due to their unique merits such as remarkable stability, excellent atom utilization and low cost. Herein, a convenient and sensitive colorimetric strategy was developed for the sensing of galactose based on Fe-N-C single-atom nanozyme (Fe-SAzyme). The Fe-SAzyme was prepared through "isolation-pyrolysis" method that exhibited intrinsic peroxidase mimicking activity, which can quickly catalyze the oxidation of 3,3',5,5'-tetramethylbenzidine (TMB) to produce blue-colored oxTMB in the presence of hydrogen peroxide (H2O2). Galactose can be oxidized by galactose oxidase (Gal Ox) to generate H2O2, and Fe-SAzyme can be utilized for quantitative colorimetric detection of galactose. A good linearity between absorbance and the galactose concentration in the range of 50-500 μM was obtained with a detection limit of (LOD) 10 μM. The Fe-SAzyme based colorimetric strategy offered a rapid, convenient and economic way for galactose quantification detection, which could be used as an alternative method for galactosemia diagnosis.Carboxylic acid metabolome plays vital roles in the study of pathological mechanisms about cancer. This study aimed to find potential biomarkers for colorectal cancer (CRC) using carboxylic acids profiling. However, the identification of much more carboxylic acids was limited due to poor ionization efficiency and lack of characteristic fragment ions. Derivatization-liquid chromatography-mass spectrometry, which contains characteristic MS/MS fragments ions, were performed for carboxylic acid metabolomics analysis in CRC serum samples. 1054 carboxylic acids were quickly and selectively identified after extraction using three characteristic fragment ions and elucidation using the most suitable CE at 30 eV. Among them, 605 carboxylic acids exhibit discriminating levels between healthy and CRC patients in training cohort. Furthermore, the differential metabolites were found to be mainly enriched in amino acid metabolism, fatty acid biosynthesis and TCA cycle by MetaboAnalyst and iPath analysis. Finally, serine, glycine, and methionine were determined as the potential biomarkers after further confirmation using validation cohort and in vitro metabolic flux analysis. The above results collectively demonstrated that a new set of carboxylic acids can be quickly and selectively discovered using characteristic fragment ions.
Read More: https://www.selleckchem.com/products/orforglipron-ly3502970.html
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