The present disclosure relates to enzymatic cascade reactions for achieving better sensitivity in detecting a target nucleic acid sequence. Several aspects of the disclosure relate to a reaction master mix comprising four major components: a CRISPR enzyme, a guide RNA (gRNA), an aptamer (e.g., an inhibiting aptamer or an activating aptamer) and a signaling (i.e., reporter) enzyme. The aptamer interacts with the signaling enzyme and forms a complex, resulting in, e.g., inhibited signaling enzyme activity. When a target is present in the reaction mix, the Cas/guideRNA system becomes activated and preferentially collaterally cleave(s) all nucleic acids in the solution, including the aptamer. Once aptamer gets cleaved, the signaling enzyme is free to produce a signal. In the presence of a substrate, such signaling enzyme activity produces a robust signal that significantly improves the limit-of-detection of other techniques in the art.

The present invention relates to the diagnostic of inflammatory diseases. The inventors described methods using NET biomarkers as diagnostic biomarkers for inflammatory diseases. COVID-19, Lupus or mCRC are used here as illustrative models for investigating an inflammatory disease. Examples in highlighting variation of the respective correlation of NET biomarkers in this invention rely on the determination of the NET main constituents: (i), DNA as determined by examining the amount of circulating DNA (cirDNA) that corresponds to the amount of NET as being degradation by-products that are released into the circulation; (ii) NE; and (iii), MPO; as well as the detection of a blood compound being indirectly associated to NET formation like the anti-cardiolipin auto-antibody. The invention provides threshold values of NE, MPO, cir-nDNA, and cir-mtDNA blood concentrations and of MNR that can be combined to diagnose/screen individuals. Thus the invention relates to a method for diagnosing a subject for an inflammatory disease comprising the steps of i) determining in a sample obtained from the subject the level of at least one marker selected in the group consisting in NET protein markers, cir-nDNA, cir-mtDNA and/or a cir-DNA fragmentation index.

Methods and analyte sensors including at least a first working electrode having a first active area thereon, and performing a dip coating operation to deposit a bilayer membrane upon the first working electrode and the first active area. The bilayer may include an inner layer having a first membrane polymer and an outer layer having a second membrane polymer, the first membrane polymer and the second membrane polymer differing from one another. The dip coating operation may comprise one or more first dips in a first membrane formulation to form the inner layer of the bilayer membrane and one or more second dips in a second membrane formulation to form the outer layer of the bilayer membrane upon the inner layer.

The purpose of the present invention is to provide a method for evaluating the clogging of a filtration membrane with a protein-containing solution, including the steps of: a) passing a protein-containing solution through a filtration membrane, b) after the step a), obtaining a filtration membrane cross-section from the filtration membrane, c) treating the protein-containing solution before the step a), the filtration membrane before the step b), or the filtration membrane cross-section after the step b) with at least one stain specific to a protein aggregate, and d) confirming the presence of the protein aggregate in the filtration membrane cross-section.

The present disclosure provides systems and processes to screen for SARS-CoV-2. This disclosure teaches specific (and different) workable ranges for starting materials in a screening process for different SARS-CoV-2 variants (e.g., the Washington isolate, Alpha variant, Gamma P.1 variant, Beta variant, Iota variant, Delta variant, Omicron BA.1 variants, etc.). As shown herein, each variant has a different combination of starting materials and incubation periods, which further demonstrates the unpredictability of success that is associated with the disclosed systems and the disclosed processes. To be clear, the general ELISA process is well known by those having skill in the art. However, what is neither well known nor intuitive are the specific parameters associated with different process steps within ELISA. Those specific parameters are the subject of this disclosure.

A method of assaying a heat treated sample containing probiotic bacteria to enumerate viable culturable probiotic bacteria in the heat treated sample, comprises: providing a suspension comprising the heat treated sample and a nutrient broth; optionally, incubating the suspension; adding an aliquot of the incubated suspension and agar to a culture plate by a pour plate or spread plate method to provide a culture medium; incubating the culture medium for a period of time to allow probiotic bacterial colonies to grow in and/or on the culture medium; and enumerating the probiotic bacterial colonies, wherein the culture medium comprises added catalase and a pyruvate salt.

A method and apparatus for labeling a tissue section is provided. In certain aspects, the methods comprise labeling a tissue sample via a plurality of immunohistochemistry (IHC) assays for detection of markers for characterization of a cancer origin in an individual having a cancer of unknown primary (CUP). The disclosed IHC assays employ chromogen-based detection methods for improved sample efficiency and visualization of biomarkers. Further disclosed is an apparatus for carrying out the disclosed methods.

A reagent kit, staining method, and application for combining dual immunohistochemistry with elastin fiber staining. The reagent kit includes a combination of primary antibodies, consisting of anti-CK7 and anti-CD34 monoclonal antibodies from different species, such as rabbit anti-human CK7 monoclonal antibody and mouse anti-human CD34 monoclonal antibody. By selecting CK7 and CD34 as the primary antibody combination, the growth pattern of tumor epithelium can be accurately indicated, and the proliferation and thickening of alveolar septal fibrous tissue caused by tumors can be clearly revealed, as opposed to structural changes of alveoli induced by other factors. Combined with independent Victoria blue staining, the elastin fibers of the walls of arteries and veins, as well as the pleura of the lungs, can be clearly visualized, aiding in the assessment of the integrity of elastin fibers on the lung surface and in determining the extent of tumor infiltration.

The present disclosure provides modulators of proteasome dynamics and/or function in a mammalian cell, compositions and uses thereof. The disclosed modulating compounds are characterized by affecting at least one of: mammalian target of rapamycin (mTOR) activation and/or lysosomal association, proteasome cellular localization, the activity and/or level/s and/or the post translational modification/s (PTM/s), and/or subcellular localization of at least one signaling molecule participating directly or indirectly in at least one pathway mediating said proteasome dynamics/function.

A hybridoma cell line secreting an ActA monoclonal antibody and a use thereof. The hybridoma cell line was deposited on Jul. 8, 2021, at the China Center for Type Culture Collection (CCTCC) with the depository accession number of CCTCC NO: C2021174. The present invention also discloses the ActA monoclonal antibody secreted by this hybridoma cell line or its progeny cell line, a detection kit containing this ActA monoclonal antibody, immunomagnetic beads, the preparation method and use of immunomagnetic beads, and competitive ELISA and indirect ELISA detection methods. The ActA monoclonal antibody of the present invention has the advantages of high titer, good specificity, and strong affinity with the natural antigen. The Listeria competitive ELISA detection kit and immunomagnetic beads developed based on this antibody have high sensitivity and good stability, effectively monitoring the level of ActA antibodies in clinical serum samples, and can be used for labeling Listeria monocytogenes.