56 results about "Methylumbelliferone" patented technology

A method is provided for determining the activity of an enzyme which releases methylumbelliferone (MU) from an MU-containing substrate wherein the enzyme has a pH optimum below the pKa of MU comprising: contacting a sample suspected of containing the enzyme with the MU-containing substrate at a pH suitable for activity of the enzyme to allow release of MU by the enzyme; contacting the sample with light of a wavelength in the range of about 310 nm to about 350 nm; determining fluorescence produced by the released MU, thereby determining the activity of the enzyme. This real time method provides improved diagnostic methods, for example for diseases associated with an abnormal level of activity of a glycosidase. The real time assay also can be used to screen compounds for their ability to modulate enzyme activity using MU-containing substrates.

The invention provides self-repairing polyurethane, and a preparation method and a self-repairing method thereof. The preparation method comprises the following steps: (1) subjecting diisocyanate andpolyether glycol to a prepolymerization reaction so as to obtain a prepolymer mixed liquid; (2) subjecting 4-methylumbelliferone to cross-linking under the irradiation of ultraviolet light to obtain adimer of 4-methylumbelliferone; (3) adding the dimer obtained in the step (2) to the prepolymer mixed liquid obtained in the step (1), carrying out polymerization, and then performing post-treatmentto remove a solvent and unreacted reactants so as to obtain the self-repairing polyurethane. The polyurethane material obtained by using the preparation method provided by the invention has a molecular weight of 53,000 or more, narrow molecular weight distribution (which is in a range of 1.1 to 1.48), good mechanical properties (with tensile strength being 17 to 28 MPa and elongation at break being 590 to 920%), and high self-repairing efficiency (55% or more).

A method for the synthesis of a compound of formula (1) or a salt thereof, wherein A is a carbohydrate linker which is a lactosyl moiety or which consists of a lactosyl moiety and at least one monosaccharide unit selected from the group consisting of: glucose, galactose, N-acetylglucosamine, fucose and N-acetyl neuraminic acid; and wherein R1 is one of the following anomeric protecting groups: a) -OR2, wherein R2 is a protecting group removable by catalytic hydrogenolysis; b) -SR3, wherein R3 is an optionally substituted alkyl, an optionally substituted aryl or an optionally substituted benzyl; c) -NH- C(R")=C(R')2, wherein each R' independently is one of the following electron withdrawing groups: -CN, -COOH, -COO-alkyl, -CO-alkyl, -CONH2, -CONH- alkyl or -CON(alkyl)2, or wherein the two R'-groups are linked together and form -CO-(CH2)2-4-CO- and thus form, together with the carbon atom to which they are attached, a 5-7 membered cycloalkan-1,3-dione, in which dione any of the methylene groups is optionally substituted with 1 or 2 alkyl groups, and R" is H or alkyl, in which a fucosyl donor of formula (2) wherein X is selected from the group consisting of: a guanosine diphosphatyl moiety, a lactose moiety, azide, fluoride, optionally substituted phenoxy-, optionally substituted pyridinyloxy-, optionally substituted 3-oxo-furanyloxy- of formula (A), optionally substituted 1,3,5-triazinyloxy- of formula (B), 4-methylumbelliferyloxy-group of formula (C), and a group of formula (D) wherein Ra is independently H or alkyl, or two vicinal Ra groups represent a=C(Rb)2 group, wherein Rb is independently H or alkyl, Rc is independently selected from the group consisting of alkoxy, amino, alkylamino and dialkylamino, Rd is selected from the group consisting of H, alkyl and -C(=O)Re, wherein Re is OH, alkoxy, amino, alkylamino, dialkylamino, hydrazino, alkylhydrazino, dialkylhydrazino or trialkylhydrazino, is reacted with an acceptor of formula H-A-R1 or a salt thereof, wherein A and R1 are as defined above, under the catalysis of an enzyme capable of transferring fucose. A compound of formula 1', its use in manufacture of human milk oligosaccharides, a method of manufacture of human milk oligosaccharides, and a fucosyl donor are also provided.

The invention relates to a detection medium and a detection method for Cronobacter in food. The detection method of the present invention utilizes the activity of alpha-glucosidase produced by Cronobacter to detect Cronobacter in liquid culture medium, and can identify Cronobacter while carrying out selective enrichment. The Cronobacter broth medium of the present invention may comprise 18-20 parts by weight of mixed peptone; 3-5 parts by weight of lactose; 2-3 parts by weight of K2HPO4; 2-3 parts by weight of KH2PO4; 20-34 parts by weight of sodium chloride 0.08-0.15 parts by weight of sodium lauryl sulfate; 0.009-0.011 parts by weight of vancomycin; 0.07-0.10 parts by weight of 4-methylumbelliferone-α-D-glucopyranoside; pH6.8 ± 0.2, the best Culture temperature, 36°C. The invention also discloses a reagent kit for the above detection method. The invention has the advantages of shortening the detection time and avoiding subsequent tedious identification operation steps, thereby improving the detection efficiency of Cronobacter.

The invention provides compounds of formula (I) in which R1, R2, R3, R4 and R5 are independently selected from hydrogen and chromogenic substituents, and X is selected from the group consisting of hydroxyl; OR6 wherein R6 is selected from the group consisting of C1-C4 alkyl; and O-Me+ wherein Me+ is a cation derived from an organic or inorganic base. A preferred species of formula (I) is 4-Methylumbelliferyl myo-inositol-1-phosphate and salts thereof with an organic or inorganic base. The invention also provides fluorogenic methods for detecting various pathogenic bacteria, e.g., Listeria, Staphylococcus and Clostridium species, using substrates containing at least one formula (I) compound. A kit for detecting a phosphatidyl-inositol-specific phospholipase C enzyme as an indication of bacterial activity is disclosed.

This invention relates to a culture medium for the rapid detection of coliform and coliform groups, which is mixed from distilled water solution containing peptone, yeast extract, lactose, 4-methylumbelliferone-beta-D-galactoside, sodium chloride, dipotassium hydrogen phosphate, potassium dihydrogen phosphate, sodium lauryl sulfate and agar (or agar-free), the solution of indophenol-beta-D-glucoside and solution of cefsulodin. Peptone, yeast extract, lactose, 4-methylumbelliferone-beta-D-galactoside, sodium chloride, dipotassium hydrogen phosphate, potassium dihydrogen phosphate, sodium lauryl sulfate and agar (or agar-free) are weighed according to a certain criterion and added to distilled water for heat dissolution. The consequent solution is adjusted to a certain pH value, put into a high pressure sterilizer for sterilization and then added with filtrated and sterilized indophenol-beta-D-glucoside solution and cefsulodin solution after the temperature lowered to 50 deg.Cí‚5 deg.C. This is useful in either qualitative or quantitative detection of coliform and coliform groups in drinking water, environmental water and foods, high efficiency and accurate detection result.

The invention discloses a coli group and colibacillus testing medium which is prepared by dissolving 20 grams of peptone, 5 grams of lactose, 2 grams of bile salt, 1.5 grams of monopotassium phosphate, 4 grams of monopotassium phosphate, 5 grams of sodium chloride, 0.1 gram of 5-bromo-4-chloro-3-indole galactoside, 0.1gram of 4- methylumbelliferone glucuronide, 1 gram of aspartic acid, 0.5 gram of casamino acid, 1 gram of yeast extracts and 20 grams of agar in 1000ml of deionized water, and the coli group and colibacillus testing medium is not only used as a plate medium, can also be used as an analoids form, and is low in cost and convenient to popularize and use.

The invention relates to the technical field of pseudomonas aeruginosa detection and discloses a pseudomonas aeruginosa chromogenic culture medium and a method for quick detection by the same. The pseudomonas aeruginosa chromogenic culture medium comprises a basic medium, a chromogenic substrate and an antibacterial agent. The chromogenic substrate comprises 2,3,5-phenyl tetrazolium chloride, 4-methyl umbelliferone-D-glucuronide, and the antibacterial agent comprises nalidixic acid, cycloserine and bile salt. 2,3,5-phenyl tetrazolium chloride acts on a pseudomonas aeruginosa metabolite to showa red color; 4-methyl umbelliferone-D-glucuronide acts on pseudomonas aeruginosa to show fluorescence under a 366nm ultraviolet lamp; by joint action of 2,3,5-phenyl tetrazolium chloride and the antibacterial agent, growth of common disturbance bacteria can be inhibited, and false positive results are avoided. By adoption of the pseudomonas aeruginosa quick detection method, detection and count of pseudomonas aeruginosa can be finished in 24h, and time saving, labor saving, stability, reliability and high sensitivity are realized.

A method is provided for determining the activity of an enzyme which releases methylumbelliferone (MU) from an MU-containing substrate wherein the enzyme has a pH optimum below the pKa of MU comprising: contacting a sample suspected of containing the enzyme with the MU-containing substrate at a pH suitable for activity of the enzyme to allow release of MU by the enzyme; contacting the sample with light of a wavelength in the range of about 310 nm to about 350 nm; determining fluorescence produced by the released MU, thereby determining the activity of the enzyme. This real time method provides improved diagnostic methods, for example for diseases associated with an abnormal level of activity of a glycosidase. The real time assay also can be used to screen compounds for their ability to modulate enzyme activity using MU-containing substrates.

The invention discloses a method for synthesizing various glucosides on a basis of 4-methylumbelliferone. According to the invention, a glycosyl donor peracetyl saccharide and a glycosyl acceptor 4-methylumbelliferone are subjected to a glycosylation reaction under room temperature or under heating with dichloromethane or 1,2-dichloroethane as a solvent and with the combined effect of Lewis acid boron trifluoride ethyl ether and organic alkali triethylamine or pyridine; and protecting groups are removed, such that various glucosides based on 4-methylumbelliferone can be obtained. The glucosides include 4-methylumbelliferone-beta-D-glucopyranosiduronide, 4-methylumbelliferone-beta-D-glucopyranoside, 4-methylumbelliferone-beta-D-xylopyranoside, 4-methylumbelliferone-beta-D-ribofuranoside, 4-methylumbelliferone-alpha-D-galactopyranoside, and 4-methylumbelliferone-alpha-D-mannopyranoside. The method is simple, and can produce a beta or alpha single-configuration target. A glycosylation reaction yield can reach 17-93%.

The invention relates to a growth medium allying a fluorogenic substrate and a pH-sensitive fluorophore, in particular the combination of 4- methylumbelliferone (4-MU) and derivatives of fluorescein. This growth medium is used for the detection by fluorescence of microorganisms by coupling the fluorescence measurements relating to the pH-sensitive fluorophor and the fluorescence measurements relating to the activation of the fluorogenic substrate(s) by the microorganisms.

The invention discloses a preparation method of 4-methyl umbrella-shaped keto-beta-D-pyran galactoside. The method comprises the following steps: step 1, preparing 4-methyl umbrella-shaped ketone; step 2, preparing tetra acetyl-Alpha-D-pyran galactoside; step 3, preparing 4-methyl umbrella-shaped keto-2,3,4,6-tetra acetyl-beta-D-pyran galactoside; and step 4, preparing the 4-methyl umbrella-shaped keto-beta-D-pyran galactoside. The method is moderate in reaction condition, namely, the reactions are carried out at a room temperature; heavy metallic salts are not required, so that the environmental friendliness is realized; the yield is high, namely, the one-step yield of glycosylations can reach 73.1% and the total yield of the glycosylations can rach 32.9%. The microbiology experiment proves that the effects of the 4-methyl umbrella-shaped keto-beta-D-pyran galactoside are equal to the effects of an imported substrate, so that the 4-methyl umbrella-shaped keto-beta-D-pyran galactoside can be used for replacing an imported product.

Provided herein are methods of treatment, including methods of treating subjects having or at risk of having or having a viral infection, and specifically a SARS-CoV-2 viral infection. The methods provided include the administration of 4-methylumbelliferone (4-MU), palmitoylethanolamide (PEA), reservatrol, fisetin, H2, nebulized hyaluronidase or combinations thereof. Also provided herein are a respiratory assistance device, methods of generating a customized respiratory assistance device, methods of treating a coronavirus infection, and methods of inhibiting a coronavirus infectivity, virulence and / or spread.

The invention discloses a synthesis method for a Beta-glucuronidase precipitation type fluorometric substrate based on 2-(benzothiazole-2'-yl)-4-bromophenol. The synthesis method comprises the following three steps of reaction: (1) glycosylation reaction; (2) aromatic cyclization reaction; (3) protecting group removal reaction. The yield of each step of reaction in the synthesis method can reach the medium level or more and even up to more than 90 percent, the reaction conversion rate of materials with high prices or preparation costs is relatively high, the total yield of the three steps canreach 37 percent, moreover, the reaction condition is mild, and the synthesis method is easy to implement. Furthermore, the step of glycosylation reaction and the step of protecting group removal reaction in the synthesis method can be applied.

The invention provides a method for determining coumarin and metabolite of coumarin in mouse blood on basis of a UPLC (ultra performance liquid chromatography)-orbitrap HRMS (high-resolution mass spectrum). The method is characterized in that with 4-methylumbelliferone as an internal standard, a sample is directly subjected to XDB-C18 chromatographic column separation and mass-spectrographic analysis after protein precipitation performed through acetonitrile, and a Full Ms / ddMS<2> mode including first-level full scanning and second-level data dependent scanning is adopted during mass spectrographic analysis. Coumarin is a specific substrate of a cytochrome P450 2A5 of a mouse, and changes of CYP2A5 enzymatic activity are evaluated by determining generation of 7-hydroxycoumarin in the mouse blood with a liquid chromatography-mass spectrometry technology. The method has good selectivity, high analysis efficiency, is simple to operate, stable, quick, high in sensitivity, good in repetition and suitable for detection and analysis of coumarin and the metabolite of coumarin in the mouse blood.

The invention discloses a reagent and card for detecting organophosphorus and carbamates pesticide residues. Through including 4-methylumbelliferone ester in a molecule with a hydrophobic space, the stability is improved. The 4-methylumbelliferone ester reagent of some examples of the invention can be stably present in an aqueous solution, and can also be hydrolyzed by a reaction with cholinesterase, so that the 4-methylumbelliferone ester reagent can be used as a color developer for the rapid detection of organophosphorus and carbamates pesticide residues.

The invention discloses a detection kit for alpha-galactosidase activity, and belongs to the field of biochemical detection. The detection kit comprises a reaction solution, a precipitant, a stop solution, a linear standard substance and a reference substance, wherein the linear standard substance is a 4-methylumbelliferone solution, and the reaction solution is a 4-methylumbelliferone acyl-alpha-D-galactoside solution. The kit can rapidly and accurately carry out in-vitro detection on the activity of alpha-galactosidase in a human blood sample, and can achieve the purpose of rapidly, conveniently and accurately assisting in diagnosing Fabry disease patients in batches by detecting a collected filter paper dry blood slice sample or a venous blood sample; and the kit is high in detection sensitivity, good in selectivity, strong in anti-interference capability, good in reproducibility, simple and convenient to operate, good in product stability and easy to use clinically.

The present invention discloses a method to determine whether a warning sign will occur in a subject with the Flavivirus infectious illness, and a pharmaceutical composition. The method includes steps of measuring the level of serum hyaluronan of the subject, and determining that the warning sign will occur in his / her illness course when the level is higher than or equal to 70 ng / mL. It is identified in the invention that the level of the serum hyaluronan is an excellent predicator for the severity of the Flavivirus infectious illness. The pharmaceutical composition is used for blocking the serum hyaluronan of the Flavivirus-infected subjects to prevent inflammation, and includes a therapeutically effective amount of a 4-methyl umbelliferone sodium salt, a CD44 siRNA, an anti-CD44 antibody or a hyaluronidase, wherein the therapeutically effective amount is an effective blood concentration of the 4-methyl umbelliferone sodium salt, the CD44 siRNA, the anti-CD44 antibody, and the hyaluronidase of the subject being in a range from 0.05 mM to 5 mM, 1 nM to 100 nM, 5 μg / ml to 500 μg / ml and 0.5 unit / mL to 50 units / mL, respectively.

The invention discloses a biodegradable asynchronous response polymalic acid active deformation material, which relates to the technical field of biomaterials. Polymalic acid, polycaprolactone and 4-methylumbelliferone single-terminated polycaprolactone are used to obtain the polymer through esterification, and then with ultraviolet radiation and stretching fixation process, a kind of The biodegradable polymalic acid active deformation material with asynchronous response has superior performance, simple manufacture and convenient use.

The invention provides an anti-influenza virus compound. The compound is a benzoic acid derivative, compared with the active compound on the market, the active compound of the anti-influenza virus compound has no chiral center, and the synthesis is simpler; the raw material is simple and easy to obtain, the market supply is more, and the price is low. Through an in-vitro neuraminidase-specific fluorogenic substrate 2'-4-methylumbelliferone-a-N-acetylneuraminic acid test, the anti-influenza virus compound has strong anti-H3N2 influenza activity and strong Anti-H1N1 influenza activity, has better therapeutic effect on influenza B, and is expected to develop anti-influenza virus drugs. The preparation method of the invention is simple, and suitable for industrial production.

The invention provides a 4-methylumbelliferone enrichment material. The enrichment material is prepared by the following method: mixing agarose and a NaOH solution and dimethyl sulfoxide at the mass ratio of agarose to the NaOH solution to dimethyl sulfoxide beign 1:0.2-5:1-10, placing the mixture under the thermostatic waterbath condition, stirring while adding halogenated hydrocarbon or halogenated acid at the mass ratio of agarose to halogenated hydrocarbon or halogenated acid being 1: 0.2-5, continuously stirring after addition and carrying out a thermostatic reaction until the end of thereaction, cleaning with acetone and deionized water, centrifuging and drying to prepare the 4-methylumbelliferone enrichment material. By introducing the material to the bottom of a culture tube, simultaneous determination of Escherichia coli and Escherichia coli can be simultaneously determined in the same culture tube when respective chromogenic / fluorogenic substrate mediums of Escherichia coliand Escherichia coli are added into the culture tube for culture.

The invention discloses a biodegradable asynchronous response polymalic acid active deformation material, which relates to the technical field of biological materials. Polymalic acid, polycaprolactone and 4-methylumbelliferone single-terminated polycaprolactone are subjected to an esterification reaction to prepare the polymer, and then the biodegradable asynchronous response polymalic acid active deformation material is obtained through ultraviolet radiation and stretching fixation processes, and the biodegradable asynchronous response polymalic acid active deformation material is excellent in performance, easy to manufacture and convenient to use.

The invention discloses an ITO-based iron ion photoelectric sensor and a preparation method thereof, belonging to the technical field of metal ion detection. The ferric ion photoelectric sensor of the invention comprises two parts, a modified fluorescent material and an ITO substrate. By modifying ITO, 4-methylumbelliferone and chitosan-modified fluorescein isothiocyanate materials are cured on the surface of ITO to construct a "one-body dual-purpose" iron ion with both fluorescent and electrochemical properties sensor. Combining the visualization of fluorescence analysis and the advantages of sensitive and rapid electrochemical analysis, the fluorescence characteristics of the electrode are used to realize the visual determination of the concentration of iron ions in water. The results showed that the sensor for Fe 3+ Selective fluorescent recognition at 10‑ 1 ~10- 7 Within the range, the concentration of iron can be judged visually. Fe ion photoelectric sensor of the present invention is to Fe 3+ It also showed good electrochemical detection performance, and the Fe 3+ The detection limit can be as low as 5nM.

The invention discloses a high-efficiency method for screening salmonella and shigella in enterobacteriaceae, and the method comprises steps of: spreading a specimen on a Mac Conkey agar plate, carrying out incubation at a temperature of 37 DEG C, and selecting a colorless bacterial colony, wherein a portion of the bacterial colony is used for detecting an oxidase activity; inoculating the other portion of the bacterial colony on a filter paper strip soaked with tryptophan, pyroglutamyl-4-methoxy naphthylamines and 4-methylumbelliferone ioxynil octoate, and moistening the inoculated filter paper strip with a drop of water or a drop of buffer; putting the moistened and inoculated filter paper strip in a moisture-maintaining vessel to subject to incubation, and putting the filter paper strip under a Wood lamp to detect fluorescence, wherein fluorescence-positive indicates that a C-8 esterase activity exists; and dropping a drop of a solution containing a diazo dye and iron chloride on the filter paper strip, so that if the filter paper strip shows fluorescence and invariant color, the bacterial colony on the filter paper strip can be identified as salmonella; and if the filter paper strip shows no fluorescence but invariant color, the bacterial colony on the filter paper strip can be identified as shigella. The method of the invention can detect activities of three kinds of enzymes at the same time by employing a method of mixing substrates, the experiment method is simple, a dosage of substrates is lesser, cost is low, and salmonella and shigella can be screened out from nonpathogenic bacteria in enterobacteriaceae efficiently.

The invention discloses a reagent and a card for detecting residues of organophosphorus and carbamate pesticides. The stability of the 4-methylumbelliferyl ketone ester is clathrated in molecules with hydrophobic spaces. The 4-methylumbelliferyl ester reagent of some examples of the present invention can exist stably in an aqueous solution, and can also be hydrolyzed with cholinesterase, which makes it applicable to organophosphorus and amino groups as a color developing agent. Rapid detection of formate pesticide residues.

The invention discloses an application of 4-methylumbelliferone in preparation of antiallergic drugs, the chemical structural formula of the 4-methylumbelliferone is shown in the specification, in an in-vitro experiment, through an in-vitro research method for comparing RBL-2H3 and BMMCs degranulation and an in-vivo experiment research method for a passive sensitization model PCA and an active systemic sensitization model ASA, the application of 4-methylumbelliferone in preparation of antiallergic drugs is realized, and the application of 4-methylumbelliferone in preparation of antiallergic drugs is realized. It is determined that the 4-methylumbelliferone can relieve the degranulation effect of IgE-mediated mast cells and effectively relieve local and systemic anaphylactic reaction symptoms of an organism.

The invention provides an anti-influenza virus compound. The compound is a benzoic acid derivative, compared with the active compound on the market, the active compound of the anti-influenza virus compound has no chiral center, and the synthesis is simpler; the raw material is simple and easy to obtain, the market supply is more, and the price is low. Through an in-vitro neuraminidase-specific fluorogenic substrate 2'-4-methylumbelliferone-a-N-acetylneuraminic acid test, the anti-influenza virus compound has strong anti-H3N2 influenza activity and strong Anti-H1N1 influenza activity, has better therapeutic effect on influenza B, and is expected to develop anti-influenza virus drugs. The preparation method of the invention is simple, and suitable for industrial production.

The invention discloses a method for synthesizing various glucosides on a basis of 4-methylumbelliferone. According to the invention, a glycosyl donor peracetyl saccharide and a glycosyl acceptor 4-methylumbelliferone are subjected to a glycosylation reaction under room temperature or under heating with dichloromethane or 1,2-dichloroethane as a solvent and with the combined effect of Lewis acid boron trifluoride ethyl ether and organic alkali triethylamine or pyridine; and protecting groups are removed, such that various glucosides based on 4-methylumbelliferone can be obtained. The glucosides include 4-methylumbelliferone-beta-D-glucopyranosiduronide, 4-methylumbelliferone-beta-D-glucopyranoside, 4-methylumbelliferone-beta-D-xylopyranoside, 4-methylumbelliferone-beta-D-ribofuranoside, 4-methylumbelliferone-alpha-D-galactopyranoside, and 4-methylumbelliferone-alpha-D-mannopyranoside. The method is simple, and can produce a beta or alpha single-configuration target. A glycosylation reaction yield can reach 17-93%.