Utilization of capillary electrophoresis for screening identification of Escherichia coli in water (faecal contamination) and pharmaceuticals is described. The method uses an electrolyte 4.5 mM in Tris, 4.5 mM in H3BO3 and 0.1 mM in EDTA with 0.0125 % poly(ethylene oxide). A brief survey of the published methods for analysis of microorganisms by capillary electrophoresis is given.
Cytokinin hormones are important regulators of development and environmental responses of plants that execute their action via the molecular machinery of signal perception and transduction. The limiting step of the whole process is the availability of the hormone in suitable concentrations in the right place and at the right time to interact with the specific receptor. Hence, the hormone concentrations in individual tissues, cells, and organelles must be properly maintained by biosynthetic and metabolic enzymes. Although there are merely two active cytokinins, isopentenyladenine and its hydroxylated derivative zeatin, a variety of conjugates they may form and the number of enzymes/isozymes with varying substrate specificity involved in their biosynthesis and conversion gives the plant a variety of tools for fine tuning of the hormone level. Recent genome-wide studies revealed the existence of the respective coding genes and gene families in plants and in some bacteria. This review summarizes present knowledge on the enzymes that synthesize cytokinins, form cytokinin conjugates, and carry out irreversible elimination of the hormones, including their phylogenetic analysis and possible variations in different organisms.
Abstract Background Neurodegenerative diseases are a heterogeneous group of illnesses. Differences across patients exist in the underlying biological drivers of disease. Furthermore, cross‐diagnostic disease mechanisms exist, and different pathologies often co‐occur in the brain. Clinical symptoms fail to capture this heterogeneity. Molecular biomarker‐driven approaches are needed to improve patient identification & stratification with the aim of moving towards more targeted patient treatment strategies. Method The UK Biobank Pharma Proteomics Project has generated a proteomics dataset of unprecedented size. It consists of plasma proteomic profiles measured using the Olink 3k protein panel from over 54,000 individuals, complemented with broad phenotypic & genetic information. It is a unique resource to explore the biological correlation between neurodegenerative diseases as well as with other indications. We applied a multi‐task deep learning (MTL) approach to generate models that predict disease status based on plasma proteomics profiles for a broad range of indications, including neurodegenerative diseases such as Alzheimer’s and Parkinson’s disease. The MTL approach first learns fundamental biological patterns by knowledge sharing between diseases in the initial segment of the network. Subsequently, understanding of a specific indication is refined with dedicated training. This improves model generalizability and statistical power. The neural network also produces low‐dimensional embeddings of proteomic profiles that can be used for sample clustering and to derive insights about disease‐associated processes. Result The average precision‐recall area‐under‐the‐curve (PR‐AUC) of the MTL models across all diseases is 0.72 vs. 0.67 for the baseline single task logistic regression models. For Alzheimer’s disease, the MTL classifier has a PR‐AUC of 0.76. Feature importance scores were calculated using the SHAP method. Top features for Alzheimer’s disease included several known biomarkers (e.g., GFAP, NPTXR). A UMAP projection of all diseases using the feature importance scores clusters diseases by disease category. Sample clustering revealed biologically interpretable patient subgroups, such as a Parkinson’s cluster linked to lysosomal biology. Conclusion High performance of the MTL approach signifies good characterization of cross‐disease biology. This is corroborated by the model’s capability to produce meaningful low‐dimensional representations of plasma proteomics profiles that can be used for identification of cross‐diagnostic protein signatures and subtypes of neurodegenerative diseases. UKB application number 65851.
Electrospray ionization mass spectrometry (ESI-MS) analysis is frequently associated with noncovalent adduct formation, both in positive and negative modes. Anion binding and sensing by mass spectrometry, notably more challenging compared to cation binding, will have major research potential with the development of appropriate sensors. Here, we demonstrated identification of stable bisquaternary dication adducts with trifluoroacetate (TFA
Cytokinin dehydrogenase (CKX; EC 1.5.99.12) degrades cytokinin hormones in plants. There are several differently targeted isoforms of CKX in plant cells. While most CKX enzymes appear to be localized in the apoplast or vacuoles, there is generally only one CKX per plant genome that lacks a translocation signal and presumably functions in the cytosol. The only extensively characterized maize CKX is the apoplastic ZmCKX1; a maize gene encoding a non-secreted CKX has not previously been cloned or characterized. Thus, the aim of this work was to characterize the maize non-secreted CKX gene (ZmCKX10), elucidate the subcellular localization of ZmCKX10, and compare its biochemical properties with those of ZmCKX1. Expression profiling of ZmCKX1 and ZmCKX10 was performed in maize tissues to determine their transcript abundance and organ-specific expression. For determination of the subcellular localization, the CKX genes were fused with green fluorescent protein (GFP) and overexpressed in tomato hairy roots. Using confocal microscopy, the ZmCKX1–GFP signal was confirmed to be present in the apoplast, whereas ZmCKX10–GFP was detected in the cytosol. No interactions of ZmCKX1 with the plasma membrane were observed. While roots overexpressing ZmCKX1–GFP formed significantly more mass in comparison with the control, non-secreted CKX overexpression resulted in a small reduction in root mass accumulation. Biochemical characterization of ZmCKX10 was performed using recombinant protein produced in Pichia pastoris. In contrast to the preference for 2,6-dichlorophenolindophenol (DCPIP) as an electron acceptor and trans-zeatin, N6-(Δ2-isopentenyl)adenine (iP) and N6-(Δ2-isopentenyl)adenosine (iPR) as substrates for ZmCKX1, the non-secreted ZmCKX10 had a range of suitable electron acceptors, and the enzyme had a higher preference for cis-zeatin and cytokinin N-glucosides as substrates.
1. Utilisation of microfluidisation to enhance enzymatic and metabolic potential of lactococcal strains as adjuncts in Gouda type cheese A.B. Yarlagadda, M.G. Wilkinson, M.G. O'Sullivan, K.N. Kilcawley International Dairy Journal CrossRef
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