КОМПОЗИЦИИ И СПОСОБЫ ДЛЯ ЛЕЧЕНИЯ БОЛЕЗНИ ГОШЕ

Группа изобретений относится к определению продукции нейтрализующих антител у субъектов, проходящих лечение болезни Гоше. Раскрыт способ детекции нейтрализующего антитела против глюкоцереброзидазы в образце от субъекта, включающий иммобилизацию глюкоцереброзидазы на поверхность; приведение образца в контакт с иммобилизованной глюкоцереброзидазой; стадию промывки; добавление меченой глюкоцереброзидазы; стадию промывки для удаления меченой глюкоцереброзидазы, которая не связалась с антителом против глюкоцереброзидазы; обнаружение и количественную оценку метки; оценку присутствия специфического изотипа антитела против глюкоцереброзидазы, где изотип выбран из группы, состоящей из IgG, IgM, IgA и IgE, и определение, нейтрализует ли антитело против глюкоцереброзидазы активность глюкоцереброзидазы, с использованием клеток, которые экспрессируют человеческие рецепторы макрофагов против маннозы (MMR). Также раскрыт способ идентификации субъекта, для которого будет эффективно лечение болезни Гоше ...

БИОМАРКЕРЫ, АССОЦИИРОВАННЫЕ С ПРЕДИАБЕТОМ, ДИАБЕТОМ И СВЯЗАННЫМИ С ДИАБЕТОМ СОСТОЯНИЯМИ

Изобретение относится к области медицины и касается способа, тест-системы диагностики диабетической нефропатии. Сущность способа заключается в том, что в образце от субъекта определяют биомаркер, который представляет собой подобный антигену CD5. Использование способа позволяет диагностировать диабетическую нефропатию. 5 н. и 28 з.п. ф-лы, 13 табл., 5 ил., 1 пр.

СПОСОБ ДЕТЕКЦИИ ВАРИАНТА ПОСЛЕДОВАТЕЛЬНОСТИ НУКЛЕИНОВОЙ КИСЛОТЫ С ПОМОЩЬЮ АНАЛИЗА ТЕРМИНАЦИИ СО СДВИГОМ

Изобретение относится к молекулярной биологии и может быть использовано для выявления любой мутации в заданном сайте, известной последовательности нуклеиновой кислоты. В основе способа лежит принцип достройки затравки, комплементарной последовательности анализируемого образца, непосредственно примыкающей к нуклеотиду - мишени. При этом в реакционную смесь для достройки затравки вводят терминаторный нуклеотид того типа, который комплементарен нуклеотиду мишени в составе матрицы (указанный тип нуклеотида также может отсутствовать в реакции), и нуклеотиды трех других типов, которые не являются терминаторными и несут выявляемую метку. Прекращение реакции вследствие включения терминаторного нуклеотида свидетельствует об отсутствии мутации в исследуемом сайте; продолжение наращивания цепи за счет меченых нетерминаторных нуклеотидов - о ее наличии. Способ обеспечивает возможность простого, быстрого и экономичного определения мутаций любого типа. 35 з.п. ф-лы, 4 ил., 6 табл.

ВЫЯВЛЕНИЕ ПРОДУЦИРУЮЩИХ КАРБАПЕНЕМАЗУ ВИДОВ ENTEROBACTERIALES, PSEUDOMONAS И ACINETOBACTER С ПРИМЕНЕНИЕМ ХРОМОГЕНА

Изобретение относится к выявлению присутствия продуцирующих карбапенемазу Enterobacteriales, Pseudomonas и Acinetobacter в биологическом образце. Предложен способ выявления присутствия продуцирующих карбапенемазу Enterobacteriales, Pseudomonas и Acinetobacter в биологическом образце, включающий обеспечение образца, в котором предполагается присутствие продуцирующих карбапенемазу Enterobacteriales, Pseudomonas или Acinetobacter, реагирование указанного образца с раствором (7R)-7-[(z)-2-(2-аминотиазол-4-ил)-(Z)-2(метоксиимино)ацетамид]-3-(2,4-динитростирил)-3-цефем-4-карбоновой кислоты) или ее соли. При этом реакцию осуществляют в присутствии ингибитора β-лактамаз расширенного спектра (ESBL), ингибитора AmpC или их смеси. Выявляют изменение цвета с желтого на оранжевый/красный в реакционной среде, когда продуцирующие карбапенемазу Enterobacteriales, Pseudomonas или Acinetobacter присутствуют в исследуемом образце. При этом изменение цвета происходит в течение 1 ч реагирования образца. Предложены ...

АЦЕТИЛ-КОФЕРМЕНТ А-КАРБОКСИЛАЗА 2 КАК МИШЕНЬ ДЛЯ РЕГУЛЯЦИИ СЖИГАНИЯ ЖИРА, НАКОПЛЕНИЯ ЖИРА, ЭНЕРГЕТИЧЕСЕОГО ГОМЕОСТАЗА И ДЕЙСТВИЯ ИНСУЛИНА

... 1. Способ стимуляции окисления жирных кислот и потери в весе у индивидуума, включающий стадию ингибирования активности ацетил-КоА-карбоксилазы 2 у указанного индивидуума. 2. Способ по п.1, где указанная активность ингибируется путем введения указанному индивидууму ингибитора ацетил-КоА-карбоксилазы 2 (АСС2). 3. Способ по п.1, где указанный индивидуум страдает патофизиологическим состоянием. 4. Способ по п.3, где указанное патофизиологическое состояние выбрано из группы, состоящей из ожирения и диабета. 5. Способ снижения уровней сахара в крови индивидуума, включающий стадию введения указанному индивидууму ингибитора ацетил-КоА-карбоксилазы 2 (АСС2). 6. Способ по п.5, где указанный индивидуум страдает диабетом. 7. Трансгенная мышь, геном которой содержит гомозиготный разрыв в эндогенном гене АСС2 кодирующем изоформу ацетил-КоА-карбоксилазу 2 ацетил-КоА-карбоксилазы, где указанный разрыв приводит к инактивации указанного гена, и где указанная мышь не продуцирует какой-либо функциональной ...

СУБСТРАТ ДЛЯ ОПРЕДЕЛЕНИЯ АКТИВНОСТИ ИНТЕГРАЗЫ ВИРУСА ИММУНОДЕФИЦИТА ЧЕЛОВЕКА

... 1. Субстрат для определения активности интегразы вируса иммунодефицита человека, содержащий концевые последовательности провирусной ДНК и меченный радиоактивной меткой, отличающийся тем, что в качестве субстрата используют фрагмент провирусной ДНК ВИЧ-1, состоящий из двух параллельно ориентированных интактных LTR-последовательностей длиной 1200-1300 п. н. 2. Субстрат по п.1, отличающийся тем, что длинные концевые повторы (размер 600-650 п.н.) расположены на 5' и 3' концах провирусной ДНК. 3. Субстрат по п.1, отличающийся тем, что в качестве ДНК для встраивания используют избыток ДНК плазмиды.

КОМПОЗИЦИИ СТАБИЛИЗИРОВАННОГО ТЕТРАЗОЛИЙ-ФЕНАЗИНОВОГО РЕАГЕНТА И СПОСОБЫ ИХ ПРИМЕНЕНИЯ

... 1. Композиция реагентов, содержащая тетразолиевый краситель, феназиновый агент переноса электронов, и эффективное количество соединения группы IIIA и/или флавинового стабилизирующего агента. 2. Композиция по п.1, в которой указанный флавиновый стабилизирующий агент представляет собой FAD. 3. Композиция по п.1 или 2, в которой указанный стабилизирующий агент группы IIIA представляет собой борат или борную кислоту. 4. Композиция по пп.1, 2 или 3, в которой указанная композиция реагентов содержит аналитическую систему, продуцирующую сигнал окисления. 5. Композиция по любому из пп.1-4, где указанная композиция представляет собой жидкую композицию. 6. Композиция по любому из пп.1-4, где указанная композиция представляет собой сухую композицию. 7. Реакционноспособная индикаторная полоска, содержащая субстрат, и композицию реагентов по любому из пп.1-6. 8. Аналитическая система обнаружения или измерения, включающая (А) реакционноспособную индикаторную полоску по п.7, и (В) автоматизированный прибор ...

ЛАБИЛЬНЫЕ ЛИНКЕРЫ ДЛЯ ВЫЯВЛЕНИЯ БИОМАРКЕРА

СПОСОБЫ И УСТРОЙСТВО ДЛЯ ОБНАРУЖЕНИЯ ГЛЮТЕНОВОЙ ЧУВСТВИТЕЛЬНОСТИ И ЕЕ ДИФФЕРЕНЦИРОВАНИЯ С ЦЕЛИАКИЕЙ

СПОСОБЫ И НАБОРЫ ДЛЯ ОБНАРУЖЕНИЯ БИОТОКСИЧНЫХ И АНТИБИОТИЧЕСКИХ ОСТАТКОВ

... 1. Набор для обнаружения антибиотических остатков, содержащий: а) индуцируемые бактериальные споры, которые продуцируют индуцируемый фермент после прорастания и индукции; b) способствующее прорастанию вещество (герминант), которое запускает быстрое прорастание указанных спор; с) соединение, которое действует в качестве индуктора, который запускает продуцирование конкретного фермента, представляющего интерес, и в то же время в качестве субстрата конкретного фермента, представляющего интерес; и d) детектор активности указанного фермента на указанном субстрате. 2. Набор по п.1, где указанным ферментом является бета-лактамаза и бета-лактам действует одновременно как в качестве индуктора, так и в качестве субстрата указанного фермента, и указанный субстрат и указанный фермент первоначально отделены друг от друга. 3. Набор по п.1, где указанным детектором является сине-черный раствор комплекса крахмал-иодид-иод. 4. Набор по п.1, где указанным детектором является полоска абсорбента, пропитанная ...

НОВЫЕ ХРОМОФОРЫ/ФЛУОРОФОРЫ И СПОСОБЫ ИХ ПРИМЕНЕНИЯ

... 1. Нуклеиновая кислота, кодирующая хромофорный и/или флуоресцентный белок, характеризующийся аминокислотной последовательностью SEQ ID NO: 02, 04, 06, 08, 10, 12, 14, 16, 18, 20, 22, 24, 26 или 28 или их мутантной последовательностью; или комплементарная нуклеиновая кислота или ее миметик, гибридизующийся в жестких условиях с указанной нуклеиновой кислотой. 2. Нуклеиновая кислота по п.1, характеризующаяся последовательностью остатков, которая по существу совпадает или идентична нуклеотидной последовательности, составляющей в длину, по меньшей мере, 10 остатков из SEQ ID NO: 01, 03, 05, 07, 09, 11, 13, 15, 17, 19, 21, 23, 25 или 27. 3. Нуклеиновая кислота по п.2, обладающая сходством последовательности, равным, по меньшей мере, приблизительно 60%, с последовательностью, составляющей в длину, по меньшей мере, 10 остатков из SEQ ID NO: 01, 03, 05, 07, 09, 11, 13, 15, 17, 19, 21, 23, 25 или 27. 4. Нуклеиновая кислота по п.1, где указанная мутантная последовательность содержит, по меньшей мере ...

СПЕЦИФИЧЕСКИЙ КРАСИТЕЛЬ ДЛЯ ОКРАШИВАНИЯ КАПСУЛЫ ЛИЧИНКИ TRICHINELLA SPIRALIS ПРИ ЕЕ ИДЕНТИФИКАЦИИ ВМЫШЕЧНЫХ ТКАНЯХ ЖИВОТНОГО

Специфический краситель для окрашивания капсулы личинки Trichinella spiralis при ее идентификации в мышечных тканях животного характеризуется тем, что является раствором кристаллического йода, йодистого калия в этиловом спирте концентрацией 90-96% при следующем соотношении компонентов, вес. ч.: кристаллический йод - 1,0 йодистый калий - 1,8-2,2 этиловый спирт концентрацией 90-96% - 6-10.

Viral drug susceptibility testing

QUALITATIVE PREDICTIVE METHOD FOR DIFFERENTIAL DIAGNOSIS OF PNEUMOCOCCIC, MENINGOCOCCIC AND VIRAL MENIGITIS, DIFFERENTIAL MENINGITIS DIAGNOSTIC METHOD AND KIT

Viral drug susceptibility testing.

Flow - Controlled magnetic particle manipulation

Inventive methods and apparatus (1, 2) are useful for collecting magnetic materials in one or more magnetic fields and resuspending the particles into a dispersion medium, and optionally repeating collection/resuspension one or more times in the same or different medium, by controlling the direction and rate of fluid flow through a. fluid flow path (10). The methods provide for contacting derivatized particles with test samples and reagents, removal of excess reagent, washing of magnetic material, and resuspension for analysis, among other uses. The methods are applicable to a wide variety of chemical and biological materials that are susceptible to magnetic labeling, including, for example, cells, viruses, oligonucleotides, proteins, hormones, receptor-ligand complexes, environmental contaminants and the like.

Flow-controlled magnetic particle manipulation.

Viral drug susceptibility testing.

Viral drug susceptibility testing.

QUALITATIVE PREDICTIVE METHOD FOR DIFFERENTIAL DIAGNOSIS OF PNEUMOCOCCIC, MENINGOCOCCIC AND VIRAL MENIGITIS, DIFFERENTIAL MENINGITIS DIAGNOSTIC METHOD AND KIT

QUALITATIVE PREDICTIVE METHOD FOR DIFFERENTIAL DIAGNOSIS OF PNEUMOCOCCIC, MENINGOCOCCIC AND VIRAL MENIGITIS, DIFFERENTIAL MENINGITIS DIAGNOSTIC METHOD AND KIT

Viral drug susceptibility testing.

Flow-controlled magnetic particle manipulation.

Flow-controlled magnetic particle manipulation.

WITH PHÄNOTYPI MEDICINE RESISTANCE TRANSKRIPTASE CORRELATED MUTATION PROFILES IN HIV-1-REVERSE

PROCEDURE AND PREPARATIONS AROUND RESISTANCE AGAINST BIOLOGICAL OR CHEMICAL THERAPIES FOR OVERCOMING

Assay system for the simplified determination of NAD(P)H

An assay system for the simplified determination of NAD(P)H or of substrates or enzymes which react with formation or consumption of NAD(P)H is described and is characterized in that it contains simultaneously, immobilized in a polymeric thin layer 2, a plurality of substances which act independently of one another as electron acceptor from NAD(P)H and have different electrochemical potentials. ...

PROCEDURE FOR THE PRODUCTION OF COMPLEX OF MULTI-ENZYMATIC STORABLE ONES OF REACTION MIXTURES AND THEIR USE

PROCEDURE FOR THE PRODUCTION OF SELFINDUCING SYNTHASE

POSH NUCLEIC ACID, POLYPEPTIDE AND TO IT RELATIONSHIP PROCEDURE

METHOD TO THE DNS SEQUENZIERUNG

SCREENING AFTER CONNECTIONS, THE TGF BETA DEPENDENT ONES OF CELL SIGNALS MODIFYING ABILITY

LINKED D-PROTEIN AND PROCEDURE FOR THE IDENTIFICATION OF RECEPTOR ACTIVITY MODULATING CONNECTIONS

USE FROM BIO MARKERS TO THE PROOF OF OVARY CANCER

PHARMACEUTICALS AND PROCEDURES FOR THE HYPOXIA TREATMENT AND SCREENING PROCEDURE FOR IT

PROCEDURE AND MEANS FOR THE ENZYMATIC REGULATION OF ACETATE

PROCEDURE FOR THE IDENTIFICATION FROM CONNECTIONS TO THE THERAPY OF AGING PROCESSES THE HEART CYCLE OF THE SYSTEM

TIME TEMPERATURE INDICATOR ON ENZYME BASIS

DIAGNOSIS AND TREATMENT OF ATHEROSKLEROSE

PRODUCTION OF POLYHYDROXYALKANOATEN MIDDLE ONE CHAIN LENGTH FROM FATTY ACID BIO-SYNTHESIS WAYS

PROCEDURE FOR APOPTOSENACHWEIS

PROCEDURE FOR THE IMPROVEMENT OF THE PROTEINKONFORMATION BY REDUCING AGENTS

CREATININSENSOR CALIBRATION

PROOF OF IONS IN LIQUIDS

A SYSTEM OF THE DNA AMPLIFICATION FOR THE DETECTION OF GENETIC ILLNESSES BY A THERMALSTABLE LIGASE

AMPLIFIZIERUNG OF CYP24 AND APPLICATION FOR IT

PROCEDURE FOR THE MEASUREMENT OF HOMOCYSTEIN

PROCEDURE FOR THE QUANTITATIVE ENZYMATIC REGULATION OF ADP

TEST FOR THE STATEMENT OF THE SUSCEPTIBILITY FOR DNS ASSOCIATING DISEASES.

PRODUCTION OF THERMALSTABLE ENZYMES.

REGULATION OF POTASSIUM IONS IN LIQUIDS.

ACETYL COA CARBOXYLASE ONE OF CANDIDA ALBICANS

BIO SENSOR ALSO ON AN SIGNAL-ACTIVE SURFACE KOVALENT FIXED BIOLOGICAL MATERIAL

COMPLEX-BOUND RESTRICTORS OF ACTIVATE-CASH OF METABOLIC ENZYMES AS MOLECULAR MARKERS FOR THE DIAGNOSTICS AND THERAPY MONITORING

A CELLFREE SYSTEM FOR THE START OF THE DNA REPLIKATION

LIQUID DELIVERY DEVICE REGULARLY BY A ANALYTEN AND A MONITORING OF THE ANALYTEN

PROCEDURE FOR THE PROOF OF INTERFERON ACTIVITY

MODIFIED GREEN FLUORSZIERENDE PROTEINS

IMMUNSUPPRESSIVE PROTEIN GOALS

PROCEDURE FOR THE IDENTIFICATION OF CONNECTIONS, THE ANAPHASE PROMOTING COMPLEX RESTRAINING

PROCEDURE FOR THE SCREENEN OF SUBSTANCES THE ONE EFFECT ON INTRAZELLULÄRE TRANSLOKATION CREDIT

ENZYMATIC DETERMINATION OF THEOPHYLLINE

ISOLATING THERMOSTABLE ENZYMES FROM ENGINEERED MESOPHILES

METHODS FOR GENERATING STABILIZED LYOPHILIZED MATERIALS

Lyophilized biological reagents, such as enzymes (e.g., PCR reagents) and antibodies, are provided that include a wax component. Thus, in some aspects, a method is provided for storing a biological reagent comprising formulating the reagent into a lyophilized composition 5 including a wax component. Methods for using such lyophilized reagents are likewise provided.

Determination of ions in fluids

High throughput in vitro screening assay for modulators of nucleic acid helicases

Autoinducer synthase modulating compounds and uses therefor

Rapid quantitative analysis of proteins or protein function in complex mixtures

Method for judging early flocculation

NUCLEIC ACID SEQUENCES ENCODING LYSK FROM DIFFERENT BORRELIA STRAINS AND USES THEREOF

SUBSTRATES, DEVICES, AND METHODS FOR CELLULAR ASSAYS

Compositions and methods for normalizing assays

Methods for analyzing protein binding events

Means of examining nephropathy

METHODS FOR IDENTIFYING AND VALIDATING POTENTIAL DRUG TARGETS___

Pharmaceuticals and methods for treating hypoxia and screening methods therefor

Gads as modifiers of the p53 pathway and methods of use

METHODS AND COMPOSITIONS FOR FUNCTIONAL UBIQUITIN ASSAYS

TYPE I INTERFERON-INDUCIBLE PROTEINS TO DETECT VIRAL INFECTION

Isoliquiritigenin Suppresses human T lymphocyte activation via targeting on cysteine 46 of IKBA Kinase

The present invention provides a compound, Isoliquiritigenin, which could inhibit immune suppressive effect resulted from direct interaction with IKK-P C46A in vivo and in vitro. Isoliquiritigenin effectively inhibits the immune-suppressive effects through regulation of T lymphocytes by inhibiting T cells proliferation and division, suppressing expression of CD69 and CD 25 as well as up-regulating CD71 in a dose-dependent manner. The present invention also provides a method of screening therapeutic agents for treating autoimmune diseases.

Metalloprotein compositions

The present invention relates to compositions comprising: a polypeptide, wherein at least a portion of the polypeptide has a coiled coil structure; and a chelate comprising a chelating agent and a metal ion; and wherein the chelate is bound to at least one amino acid of the polypeptide. In a preferred embodiment the polypeptide is a silk fibroin, wherein at least a portion of said silk fibroin has a coiled coil structure.

Temperature insensitive in vivo analyte devices, methods and systems

Membrane structures are for use in analyte sensors, where the membrane structures exhibit low temperature sensitivity. In vivo analyte monitoring devices, systems and methods that are temperature insensitive to analyte permeability at least at temperatures for which the insensitive in vivo analyte monitoring devices, systems and methods are or could be exposed (SMART devices, systems and methods), such as in vivo use temperatures like room temperatures, mammalian body temperatures, and the like. The SMART membranes regulate the permeability of analyte (e.g., glucose) through the membrane at different temperatures to maintain a constant permeability over a range of temperatures, and minimize or in some instances eliminate changes in sensitivity values of the in vivo analyte sensor with which the SMART membranes are used.

Biomarkers associated with pre-diabetes, diabetes and diabetes related conditions

Abstract The invention provides biomarkers for pre-Diabetes, Diabetes and/or a Diabetes related conditions, and methods of their use, including the biomarkers in Tables 1 and 2 such as peroxiredoxin-2, complement Clq subcomponent subunit B, sulfhydryl oxidase 1 and apolipoprotein A-IV.

Renal tissue damage detection method by detecting endogenous expression of OP-1

Assay procedure and application in identification of herbicides

Histone deacetylase as target for antiprotozoal agents

Catalyzed reporter deposition

Diagnosis and treatment of viral hepatitis

Methods of measuring the concentration of an analyte in a subject

Barley with modified SSIII

Barley grain, plants and parts thereof producing same and methods of producing and using same are described, the grain comprising (a) starch (b) a first genetic variation which is (i) an induced mutation in an endogenous gene encoding a starch synthase HI (SSIII), or (ii) a transgene which encodes an RNA molecule which reduces expression of an endogenous gene encoding an SSIII, and (c) a reduced level or activity of SSIII protein relative to the level or activity in barley grain lacking the first genetic variation. The grain may further comprise a second genetic variation in a gene encoding a protein involved in starch synthesis, starch catabolism, starch phosphorylation or non-starch carbohydrate synthesis. The second genetic variation is illustratively (i) a mutation in a gene encoding a starch synthase II (SSII), in some embodiments a starch synthase Ila (SSIIa), or (ii) a transgene which encodes an RNA molecule which inhibits expression of a gene encoding an SSII, preferably an SSIIa ...

Polynucleotide encoding an activated human t-lymphocyte-derived protein related to ubiquitin conjugating enzyme

Ubiquitin conjugating enzymes

DETERMINATION OF IONS IN FLUIDS

D-enzyme compositions and methods of their use

Plant adenylosuccinate synthetase and dna coding therefor

Gyrase inhibitors and uses thereof

Synthetic receptors, libraries and uses thereof

Synthetic gene for expressing active retroviral protein in eukaryotes

Human cystatin E

Novel diagnostic marker for type 1 diabetes mellitus

The present invention provides, as a novel diagnosis marker for type 1 diabetes mellitus, a type 1 diabetes mellitus diagnostic composition comprising alanyl-tRNA synthetase, glycyl-tRNA synthetase, asparaginyl-tRNA synthetase, or tryptophanyl-tRNA synthetase, a diagnostic kit comprising the same, and a diagnostic method using the same. The composition, the kit, and the method, according to the present invention, may be used for early diagnosis and confirmed diagnosis of type 1 diabetes mellitus because type 1 diabetes mellitus can be easily diagnosed from a patient sample.

Inhibition of the activity of kinase and synthetase enzymes

The invention provides a method of inhibiting the activity of a kinase or a synthetase, the method including binding an active site of the kinase or synthetase with a deuterated imidazole moiety, thereby inhibiting the activity of the kinase or the synthetase.

METHODS OF DETECTING SLEEPINESS

The present application is directed to methods of detecting sleepiness. 1. A method for detecting sleepiness in a subject , the method comprising:(a) providing a test sample comprising a sample of bodily fluid taken from the subject;(b) determining the concentration of one or more metabolites in the test sample from (a), wherein the sample metabolites are selected from the group consisting of the metabolites in Table 1 or a combination thereof;(c) comparing the concentration of the metabolite in (b) to a threshold concentration of the metabolite;wherein a concentration of the metabolite that is significantly different in the test sample compared to the threshold concentration of the metabolite indicates sleepiness.2. The method of claim 1 , wherein the test sample is selected from the group consisting of a blood sample claim 1 , an interstitial fluid sample claim 1 , a saliva sample claim 1 , a urine sample claim 1 , and a pancreatic sample.3. The method of claim 1 , wherein the test sample is a saliva sample.4. The method of claim 1 , wherein an elevated level of the metabolite compared to a threshold concentration of the metabolite indicates sleepiness.5. The method of claim 1 , wherein a reduced level of the metabolite compared to a threshold concentration of the metabolite indicates sleepiness.6. The method of claim 1 , wherein the metabolite is oxalate.7. The method of claim 1 , wherein the metabolite is pyrophosphate.8. The method of claim 1 , wherein sleepiness is a condition selected from the group consisting of sleep deprivation claim 1 , coma claim 1 , dyssomnias claim 1 , parasomnias claim 1 , and sleep disorders.9. A method for detecting sleepiness in a subject claim 1 , the method comprising:(a) providing a test sample comprising a sample of bodily fluid taken from the subject;(b) determining the concentration of pyrophosphate in the test sample from (a);(c) comparing the concentration of pyrophosphate in (b) to a threshold concentration of ...

COMPOSITION FOR TREATING SKIN PIGMENTATION

The invention relates to a topical composition containing a bilirubin-producing plant extract. In particular, the bilirubin-producing plant extract is obtained from the genus When topically applied to skin, the composition is effective in accelerating the degradation of heme by-products such as bilirubin present in the skin. 3. The composition of wherein the extract contains components which cause accelerated degradation of bilirubin.4. The composition of wherein the extract is derived from the whole bilirubin-producing plant claim 1 , part of the plant claim 1 , the seed of the plant claim 1 , or a cell culture of the plant.5StrelitziaPhenakospermum.. The composition of wherein the bilirubin-producing plant is from the genus or6Strelitzia.. The composition of wherein the bilirubin-producing plant is from the genus7Strelitzia nicolai.. The composition of wherein the bilirubin-producing plant extract is from the species8Strelitzia nicolai.. The composition of wherein the bilirubin-producing plant extract is taken from the seeds of the bilirubin-producing plant species9Strelitzia nicolai.. The composition of wherein the bilirubin-producing plant extract is taken from the arils of the seeds of the bilirubin-producing plant10. A method for testing the bilirubin degrading ability of a bilirubin-producing plant extract comprising:a) dispersing or dissolving bilirubin in water;b) adding measured amounts of the extract of the bilirubin-producing plant extract; andc) measuring bilirubin degradation verses time using an analytical method sensitive to detecting bilirubin and its degradation by-products.11. The method of in which the amount of the bilirubin-producing plant extract is present from about 0.001% to about 20%.12. The method of wherein the bilirubin degradation is measured using either UV claim 10 , IR claim 10 , or HPLC.13. The method of in which the analytical technique is HPLC.14. A method for accelerating the degradation of bilirubin accumulated in the skin ...

STERILIZATION INDICATORS INCLUDING A NEUTRALIZER AND METHODS

Sterilization indicators that include a neutralizer, such indicators useful for testing the effectiveness of a sterilization procedure by measuring the activity of an active enzyme whose activity is correlated with the survival of microorganisms. 120-. (canceled)22. The sterilization indicator of claim 21 , wherein the neutralizer is a sulfur-containing compound.23. The sterilization indicator of claim 22 , wherein the sulfur-containing compound is selected from the group consisting of methionine claim 22 , L-cysteine claim 22 , D-ethionine claim 22 , S-methyl-L-cysteine claim 22 , S-benzyl-L-cysteine claim 22 , sodium thiosulfate claim 22 , glutathionine claim 22 , L-cystathionine claim 22 , N-acetyl-L-cysteine claim 22 , carboxymethylcysteine claim 22 , D claim 22 ,L-homocysteine claim 22 , D claim 22 ,L-homocysteine-thiolactone claim 22 , thiodipropionic acid claim 22 , and combinations thereof.24. The sterilization indicator of claim 21 , wherein the neutralizer is a non-sulfur-containing compound.25. The sterilization indicator of claim 21 , wherein the neutralizer is isolated from the sterilant during sterilization.26. The sterilization indicator of claim 21 , wherein the carrier comprises a material in a sheet form.27. The sterilization indicator of claim 21 , wherein the carrier comprises a porous carrier and the biological material is distributed within the carrier.28. The sterilization indicator of claim 27 , wherein the porous carrier occupies at least 5% of the volume of the container in which it is located.29. The sterilization indicator of claim 27 , wherein the porous carrier occupies no more than 50% of the volume of the container.30. The sterilization indicator of claim 28 , wherein the porous carrier has a solidity of greater than 5%.31. The sterilization indicator of claim 28 , wherein the porous carrier has an effective fiber diameter of greater than 10 microns.32. The sterilization indicator of claim 21 , wherein the carrier retains residual ...

RATIONAL ENZYME MINING

A method for rational mining for induced enzymes in microbial communities is described. The method is characterized in that a community of microorganisms is provided and that the microbial populations of the community are cultivated in a container under conditions of choice, where the microorganisms are given a defined culturing medium, to eliminate matter deriving from the natural habitat and to allow the microbial community to reach a metabolic steady-state. The method is further characterized in that at least one fraction of microorganisms is taken from the container and transferred into at least two separate containers, where at least one of the fractions of microorganisms is provided with a defined medium which includes an inducing substance and/or a substance against which enzyme/enzymes is/are desired, to induce regulation of expression of the desired enzyme/enzymes, and at least one fraction is provided with a defined medium without said inducing substance and without said substance against which enzyme/enzymes is/are desired, for the purpose of comparison, and the fractions are maintained and/or cultivated. Samples of the two fractions are then withdrawn and analyzed for identification of the induced enzyme/enzymes. 1. A method for rational mining for induced enzyme/enzymes in microbial communities , comprising the steps of:(a) providing at least one community of microorganisms,(f) cultivating the microbial populations of said at least one community in at least one container under conditions of choice, where the microorganisms are given a defined culturing medium to eliminate matter deriving from the natural habitat and to allow the microbial community to reach a metabolic steady-state,(g) removal from at least one container of step (b) of at least one fraction of microorganisms, and transfer of said at least one fraction into at least two separate containers,(h) providing at least one of the fractions of microorganisms from step (c) with a defined medium ...

Ubiquitin ligase assays and related reagents

The disclosure provides, inter alia, methods and reagents for use in measuring the attachment of ubiquitin and ubiquitin-like proteins to a target protein, particularly an E3 protein.

Methods for identifying modulators of murine double minute 2

The disclosure is directed to methods of identifying compounds that modulate murine double-minute 2. More particularly, the disclosure is directed to methods of identifying compounds that modulate murine double-minute 2 neddylation activity.

Proximity Assay for In Situ Detection of Targets

A proximity detection method is described that utilizes enzymatic biotinylation to detect targets in a sample, particularly formalin fixed paraffin embedded samples using automated staining platforms. One disclosed embodiment comprises contacting the sample with a first conjugate comprising a biotin ligase and a first specific binding moiety that binds proximally to the first target; contacting the sample with a second conjugate comprising a biotin ligase substrate and a second specific binding moiety that binds proximally to the second target; subjecting the sample to conditions that allow biotinylation of the biotin ligase substrate by the biotin ligase when the first target and the second target have a proximal arrangement; and detecting biotinylation of the biotin ligase substrate. The conditions that allow biotinylation of the substrate include addition of biotin and ATP. The method also may comprise contacting the sample with a streptavidin-enzyme conjugate. Signal amplification also can be used. 1. A method for analyzing a sample to determine whether a first target is proximal to a second target , the method comprising:contacting a sample with a first conjugate comprising a biotin ligase and a first specific binding moiety for associating with a first target;contacting the sample with a second conjugate comprising a biotin ligase peptide substrate and a second specific binding moiety for associating with a second target under conditions that allow biotinylation of the peptide substrate; anddetecting the biotin if the first target is proximal to the second target.2. The method according to claim 1 , wherein labeling the first target includes contacting the sample with a first primary antibody specific to the first target and labeling the second target includes contacting the sample with a second primary antibody specific to the second target.3. The method according to claim 2 , wherein the first primary antibody is labeled with a first hapten and labeling the ...

Method and system for determining a biological response of a target to a soluble candidate substance

A method for determining a biological response of a target () to a soluble candidate substance comprises the steps: 113.-. (canceled)14. A method for determining a biological response of a target to a soluble candidate substance , the method comprising the following steps:introducing a soluble candidate substance into a laminar flow of a buffer liquid flowing through a dispersion channel to form a candidate substance solute in the buffer liquid having an initial concentration profile;by the laminar flow of the buffer liquid through the dispersion channel dispersing the initial concentration profile of the candidate substance solute in the buffer liquid to form a dispersed concentration profile of the candidate substance solute in the buffer liquid;directing the laminar flow of the buffer liquid containing the candidate substance solute having the dispersed concentration profile into a detection channel to form a final symmetrical concentration profile of the candidate substance solute in the buffer liquid in the detection channel;introducing a target into the detection channel in a manner so as to obtain a combined concentration profile in the buffer liquid, the combined concentration profile comprising a constant target concentration profile overlying the final symmetrical concentration profile of the candidate substance solute;holding in the detection channel at least one half of the combined concentration profile contained in the buffer liquid; andoptically scanning the at least one half of the combined concentration profile contained in the buffer liquid held in the detection channel to detect at the various concentrations of the candidate substance solute of the combined concentration profile an optical signal which is representative of the biological response of the target to the soluble candidate substance.15. A method according to claim 14 , wherein the step of holding in the detection channel the at least one half of the combined concentration profile ...

Method and apparatus utilizing enzyme substrates producing slow diffusing fluorescent product appearances and chromogens, and use in combination with fast diffusing product appearances

A new method of rapid detection of cells, microorganisms, or other items is described using various combinations of indicator enzyme substrates which can be used to yield fluorophoric and chromophoric appearances due to enzymatic activity. One aspect of the invention is the use of a family of compounds that can be used to produce slow diffusing fluorophoric appearances. Another aspect is a family of compounds identified as dual enzyme substrates that can be used to produce both fluorophoric and chromogenic appearances. 1. A method of detecting an enzyme reaction , using an enzyme substrate diagnostic material that can be used to provide for at least two different detectable qualities , wherein at least one of these qualities is slow fluorescent if it is the only quality being monitored for detection.2. A method of detecting an enzyme reaction according to claim 1 , comprising the steps of:Obtaining a sample,Contacting said sample with an enzyme substrate diagnostic material that can be used to provide for at least a fluorogenic quality and a chromogenic quality that each can be used to identify said enzyme reaction,Monitoring results of said contacting to detect at least presence or absence of fluorogenic quality, said presence t representing said enzyme reaction, andMonitoring the result of said contacting to detect at least presence or absence of chromogenic quality said presence representing said enzyme reaction.3. The method according to wherein the enzyme substrate is an indolyl-based enzyme substrate.4. The method according to wherein the enzyme substrate is chosen from fluorescein and resorufin based enzyme substrates.5. A method of detecting an enzyme reaction according to comprising the steps of:a. Obtaining a sample,b. Contacting said sample with an enzyme substrate diagnostic material that can be used to provide for at least a fluorogenic quality and a chromogenic quality that each can be used to identify said enzyme reaction, and a different enzyme ...

Glutamine Synthetase Reaction and Method for Quantifying Ammonia Utilizing the Same

A reagent for glutamine synthetase reaction comprising a chelating agent and glutamine synthetase, and a reagent for quantification of ammonia comprising a chelating agent, ATP, glutamic acid, glutamine synthetase, glucose, an oxidized NAD compound, ADP-dependent hexokinase, and glucose-6-phosphate dehydrogenase, are provided. 1. A method for carrying out glutamine synthetase reaction , comprising carrying out glutamine synthetase reaction in a reaction system containing ammonia , adenosine triphosphate (ATP) , and L-glutamate , wherein said reaction system further contains a chelating agent.2. The method according to claim 1 , wherein the chelating agent is a compound containing not less than four carboxyl groups in its molecular structure.3. The method according to claim 2 , wherein the chelating agent is selected from the group consisting of ethylenediaminetetraacetic acid (EDTA) claim 2 , diethylene triamine-N claim 2 ,N claim 2 ,N′ claim 2 ,N″ claim 2 ,N″-pentaacetic acid (DTPA) claim 2 , and trans-1 claim 2 ,2-diaminocyclohexane-N claim 2 ,N claim 2 ,N′ claim 2 ,N′-tetraacetic acid (CyDTA).4. A method for quantifying ammonia claim 1 , comprising providing an ammonia-containing sample; carrying out glutamine synthetase reaction using the ammonia-containing sample by the method according to to generate adenosine diphosphate (ADP); and quantifying ammonia based on the amount of ADP produced or the amount of reaction product produced from the ADP.5. The method according to claim 4 , said method further comprises:allowing glucose and ADP-dependent hexokinase to act on the ADP to produce glucose-6-phosphate,allowing glucose-6-phosphate dehydrogenase to act on the glucose-6-phosphate and an oxidized nicotinamide adenine dinucleotide (NAD) compound to produce a reduced NAD compound, and thenquantifying the reduced NAD compound or the amount of reaction product produced from the reduced NAD compound.6. The method of quantifying ammonia according to claim 5 , said ...

BIOLOGICAL PRODUCTION OF ß-LACTONES

Methods of using β-lactone biosynthetic enzyme genes and host cells expressing one or more of those genes, e.g., heterologous expression, are provided. 1. A method to prepare β-lactones in vitro , comprising:combining one or more acyl CoA substrates, one or more activated acyl substrates, one or more carboxylic acid substrates, or one or more fatty acid substrates with OleA or a homolog thereof, OleC or a homolog thereof, and OleD or a homolog thereof but not OleB, under conditions that yield one or more oxetan-2-ones.2. The method of wherein one or more 3-hydroxy acid substrates are combined with GleC or a homolog thereof claim 1 , but not OleD or a homolog thereof or OleB or a homolog thereof that is enzymatically active in the decarboxylation of oxetan-2-ones.3. The method of wherein one or more acyl CoA substrates claim 1 , one or more carboxylic acid substrates claim 1 , or one or more fatty acid substrates are combined with OleA or a homolog thereof claim 1 , OleC or a homolog thereof and OleD or a homolog thereof but not OleB or a homolog thereof that is enzymatically active in the decarboxylation of oxetan-2-ones.4. The method of wherein the one or more acyl CoA substrates are prepared by combining one or more carboxylic acids claim 1 , CoA and a ligase.5. The method of wherein the OleA or homolog thereof claim 1 , the OleD or homolog claim 1 , thereof or the OleC or the homolog thereof or any combination thereof claim 1 , are expressed in a heterologous cell.6. The method of wherein the heterologous cell is a bacterial cell claim 5 , a fungal cell claim 5 , or a yeast cell.7. The method of wherein the OleC or homolog thereof is isolated OleC or the homolog thereof claim 1 , the OleA or homolog thereof is isolated OleA or the homolog thereof claim 1 , or the OleD or the homolog thereof is isolated OleD or the homolog thereof.8. The method of wherein the combining yields a plurality of distinct oxetan-2-ones claim 1 , an oxetan-2-one or a plurality of ...

METHOD OF MAKING A MICROFLUIDIC DEVICE

A microfluidic device allowing for multiple discrete reactions sites and allowing for sequential reactions and sample analysis along with methods for device fabrication and use is provided. The microfluidic device provides a micro-total analysis system on a single substrate and has multiple reaction sites allowing for cascade reactions and analysis on a single chip using micro-quantities of sample and reagents. The microfluidic device provides discrete sites for immobilization of cognitive agents including enzymes, binding proteins, nucleic acids and the like as well as methods for quantitative analysis. The invention also provides methods for the fabrication of the device.

PROBES AND ASSAYS FOR MEASURING E3 LIGASE ACTIVITY

Provided herein is technology relating to the biological process of protein ubiquitination and particularly, but not exclusively, to compositions and methods for studying protein ubiquitination and developing therapeutics to modulate protein ubiquitination. 1. A composition comprising a ubiquitin C-terminal thioester fluorophore.2. The composition of wherein the ubiquitin C-terminal thioester fluorophore comprises a ubiquitin covalently attached to a fluorophore by a thioester.3. The composition of wherein the ubiquitin C-terminal thioester fluorophore comprises a ubiquitin covalently attached to a linker and a linker covalently attached to the fluorophore.4. The composition of wherein the fluorophore is Fluorescein claim 1 , Rhodamine claim 1 , BODIPY claim 1 , Alexa Fluor 488 claim 1 , Oregon Green 488 claim 1 , or Alexa Fluor 594.5. The composition of wherein the linker is an alkyl claim 3 , cycloalkyl claim 3 , aryl claim 3 , heteroaryl claim 3 , heteroalkyl polymer claim 3 , carbon nanotube claim 3 , quantum dot claim 3 , or nanoparticle.6. The composition of wherein the ubiquitin comprises amino acids 1-76 of the ubiquitin polypeptide (SEQ ID NO: 1).7. The composition of further comprising an E3 ligase.8. The composition of wherein the E3 ligase is NEDD4 claim 7 , NEDD4L claim 7 , ITCH claim 7 , WWP1 claim 7 , WWP2 claim 7 , SMURF1 claim 7 , SMURF2 claim 7 , NEDL1 claim 7 , NEDL2 claim 7 , E6AP claim 7 , HECTD2 claim 7 , KIAA0614 claim 7 , TRIP12 claim 7 , G2E3 claim 7 , EDD claim 7 , HACE1 claim 7 , HECTD1 claim 7 , UBE3B claim 7 , UBE3C claim 7 , KIAA0317 claim 7 , HUWE1 claim 7 , HECTD3 claim 7 , HERC1 claim 7 , HERC2 claim 7 , HERC3 claim 7 , HERC4 claim 7 , HERC5 claim 7 , HERC6 claim 7 , SopA claim 7 , NleL claim 7 , ARIH1 claim 7 , ARIH2 claim 7 , CUL9 claim 7 , ANKIB1 claim 7 , PARK2 claim 7 , RNF144A claim 7 , RNF144B claim 7 , RBCK1 claim 7 , RNF19A claim 7 , RNF19B claim 7 , RNF31 claim 7 , RNF216 claim 7 , RNF14 claim 7 , RNF217 claim 7 , or a NEL ...

E3 UBIQUITIN LIGASE (UBE3A) PROTEIN TARGETS

The invention relates to UBE3A protein targets and their usage as target engagement biomarkers for compounds that modulate ube3a expression. 1. A method for measuring UBE3A protein expression modulation in a tissue sample comprising the steps:a) providing a tissue sample of an animal or cell culture which has been treated with a UBE3A modulator,b) measuring a protein expression level in the sample of step a) of at least one protein selected from the group consisting of: CCDC88A, DST, FAM127A, FAM127B, FAM127C, PEG10 and TCAF1, PPID.c) comparing the protein expression level of the at least one protein measured in step b) to the protein expression level of the at least one protein in a control, wherein a modulated protein expression level of the at least one protein measured in step b) compared to the protein expression level of the at least one protein in the control is indicative for UBE3A protein expression modulation.2. The method of claim 1 , wherein the protein expression level of the protein measured in step b) inversely correlates to the UBE3A protein expression level.3. The method of for measuring UBE3A protein expression induction in a tissue sample comprising the steps:a) providing a tissue sample of an animal or cell culture which has been treated with a UBE3A inducer,b) measuring a protein expression level in the sample of step a) of at least one protein selected from the group consisting of: CCDC88A, DST, FAM127A, FAM127B, FAM127C, PEG10, TCAF1 and PPID.c) comparing the protein expression level of the at least one protein measured in step b) to the protein expression level of the at least one protein in a control, wherein a decreased protein expression level of the at least one protein measured in step b) compared to the protein expression level of the at least one protein in the control is indicative for UBE3A protein expression induction.4. A method for determining UBE3A target engagement of an UBE3A modulator comprising the steps:a) providing a tissue ...

REAGENTS AND METHODS FOR BIOORTHOGONAL LABELING OF BIOMOLECULES IN LIVING CELLS

Tetrazine non-canonical amino acids, methods for genetic encoding proteins and polypeptides using the tetrazine amino acids, proteins and polypeptides comprising the tetrazine amino acids, and compositions comprising the proteins and polypeptides having at least one post-translational modification thereof comprising in vivo reaction with the incorporated tetrazine amino acid and a second molecule. 6. The compound of claim 1 , wherein Ris selected from methyl claim 1 , ethyl claim 1 , n-propyl claim 1 , i-propyl claim 1 , n-butyl claim 1 , s-butyl claim 1 , t-butyl claim 1 , n-pentyl claim 1 , and n-hexyl.7. The compound of claim 1 , wherein Ris selected from trifluoromethyl claim 1 , 2-fluoroethyl claim 1 , and methoxy.8. The compound of claim 1 , wherein phenylene is 1 claim 1 ,4-phenylene.9. The compound of claim 1 , wherein phenylene is 1 claim 1 ,3-phenylene.1012-. (canceled)1416-. (canceled)17. The compound of selected from the group consisting of 4-(6-methyl-s-tetrazin-3-yl)phenylalanine (Tet-v2.0-methyl) claim 1 , 4-(6-ethyl-s-tetrazin-3-yl)phenylalanine (Tet-v2.0-ethyl) claim 1 , 4-(6-isopropyl-s-tetrazin-3-yl)phenylalanine (Tet-v2.0-isopropyl) claim 1 , and 4-(6-butyl-s-tetrazin-3-yl)phenylalanine (Tet-v2.0-n-butyl).18. The compound of selected from the group consisting of 3-(6-methyl-s-tetrazin-3-yl)phenylalanine (Tet-v3.0-methyl) claim 5 , 3-(6-ethyl-s-tetrazin-3-yl)phenylalanine (Tet-v3.0-ethyl) claim 5 , 3-(6-isopropyl-s-tetrazin-3-yl)phenylalanine (Tet-v3.0-isopropyl) claim 5 , 3-(6-t-butyl-s-tetrazin-3-yl)phenylalanine (Tet-v3.0-t-butyl) claim 5 , and 3-(6-n-butyl-s-tetrazin-3-yl)phenylalanine (Tet-v3.0-n-butyl).19. A method for making a protein or a polypeptide of interest claim 5 , comprising:{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, 'incorporating a compound of into a protein or polypeptide.'}20. A method for genetically encoding a protein or a polypeptide of interest claim 5 , comprising:{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, ' ...

HIGH THROUGHPUT ASSAY FOR MONITORING AMP PRODUCTION AND AMINOACYL-tRNA SYNTHETASE ACTIVITY

A method of conducting an enzymatic reaction assay involving adenosine 5′-monophosphate (AMP) comprising the steps of reacting one or more compounds and producing AMP, deaminating the AMP to produce IMP, and oxidating the IMP by NAD+ to produce XMP and NADH. 1. A method of conducting an enzymatic reaction assay involving adenosine 5′-monophosphate (AMP) comprising the steps of:reacting one or more compounds and producing AMP;deaminating the AMP to produce IMP; andoxidizing the IMP by NAD+ to produce XMP and NADH.2. The method of further comprising the steps of catalyzing the deamination of the AMP with AMP deaminase.3. The method of further comprising the steps of catalyzing the oxidation of the IMP with IMP dehydrogenase4. The method of further comprising the step of monitoring an absorbance of the assay at 340 nm.5. The method of further comprising the step of coupling a production of the NADH to a bacterial luciferase catalyzed oxidation of a low molecular weight aldehyde.6. The method of further comprising the step of monitoring an activity of the one or more enzymes coupled to the production of the AMP.7. The method of wherein the reaction involves one or more of aminoacyl-tRNA synthetases claim 1 , DNA ligases claim 1 , ubiqutin and ubiquitin-like ligases claim 1 , cAMP phosphodiesterases claim 1 , polyA deadenylases claim 1 , and ribonucleases.8. The method of further comprising the step of aminoacylation of a tRNA by an amino acid.9. The method of further comprising the step of substantially continuously recycling the tRNA.10. The method of further comprising the steps of recycling the tRNA using one of an editing domain claim 9 , an editing protein claim 9 , an editing protein containing an editing domain claim 9 , cyclodipeptide synthases claim 9 , and aminoacyl-tRNA using enzymes claim 9 , such as Penicillin Resistance Factor MurM and FEM ligases.11. The method of further comprising the step of one of removing and converting the amino acid from the assay ...

SUMOylation Assay and Related Reagents

Provided herein and methods and kits for identifying inhibitors of SUMOylation. The provided methods include contacting a candidate agent with (i) a SUMO-conjugating peptide comprising an N-terminal linker and a detectable tag, and (ii) a SUMO comprising a metal complex, under conditions that allow binding between the SUMO-conjugating peptide and the SUMO, and detecting the detectable tag thereby determining the level of binding between the SUMO-conjugating peptide and SUMO. A reduced level of binding as compared to the level of binding in the absence of the candidate agent indicating the agent inhibits SUMOylation. 1. A method of identifying an inhibitor of SUMOylation comprising:(a) contacting a candidate agent with (i) a SUMO-conjugating peptide comprising an N-terminal linker and a detectable tag, and (ii) a SUMO comprising a metal complex, under conditions that allow binding between the SUMO-conjugating peptide and the SUMO; and(b) detecting the detectable tag thereby determining the level of binding between the SUMO-conjugating peptide and SUMO;a reduced level of binding as compared to the level of binding in the absence of the candidate agent indicating the agent inhibits SUMOylation.2. The method of claim 1 , wherein the detectable tag is a fluorescent molecule.3. The method of claim 1 , wherein the detectable tag is biotin.4. The method of claim 3 , wherein the detecting comprises adding streptavidin labeled with a fluorescent molecule.5. The method of claim 1 , wherein the contacting further comprises a SUMO-conjugating enzyme.6. The method of claim 1 , wherein the SUMO-conjugating enzyme is selected from the group consisting of an E1 enzyme claim 1 , an E2 enzyme claim 1 , an E3 enzyme or a combination thereof.7. The method of claim 1 , wherein the metal complex is a metal chelate or a metal cryptate.8. The method of claim 7 , wherein the metal is a lanthanide metal.9. The method of claim 8 , wherein the lanthanide metal is europium or terbium.10. The ...

METHOD FOR CONTROLLING CANCER METASTASIS OR CANCER CELL MIGRATION BY MODULATING THE CELLULAR LEVEL OF LYSYL TRNA SYNTHETASE

The present invention relates to a novel function of lysyl tRNA synthetase, that is, lysyl tRNA synthetase interacts with 67LR through translocation of KRS into plasma membrane, and so enhances tumor (or cancer) cell migration, thereby having an effect on cancer metastasis. More specifically, it relates to method for controlling cancer metastasis or cancer cell migration by modulating an cellular level of lysyl tRNA synthetase, an use of an expression vector comprising a construct inhibiting KRS expression for preventing or treating cancer, an use of an agent inhibiting KRS activity for preventing or treating cancer, a method for screening an agent which modulates cancer metastasis or cancer cell migration, a method for screening an agent inhibiting an interaction between KRS and 67LR. Accordingly, cancer metastasis and cancer cell migration may be controlled using the inventive KRS, further the cellular metabolism related to laminin receptor (67LR) of plasma membrane may be controlled. The relationship between KRS and laminin receptor disclosed in the present invention may be very useful for treatment, prevention and/or diagnosis of various disease related to thereof. 1(a) contacting a testing agent with KRS and laminin receptor (67LR) in the presence of the testing agent; and(b) testing whether the selected agent regulates an interaction between KRS and laminin receptor.. A method for screening an agent inhibiting an interaction between KRS and 67LR comprising: This application is a divisional application of application Ser. No. 13/059,006 filed on Feb. 14, 2011, which is a national phase of International Application No. PCT/KR2008/004785 filed on Aug. 18, 2008. The applications are incorporated herein by reference.The present invention relates to a novel function of lysyl tRNA synthease. More specifically, it relates to a method for controlling cancer metastasis or cancer cell migration by modulating an cellular level of lysyl tRNA synthetase, an use of an ...

Quantification Method for Ammonia, Quantification Reagent Kit, Test Piece, and Ammonia Quantification Device

A method of quantifying ammonia, which method includes: performing a first reaction in which a test liquid containing ammonia is reacted with ATP and L-glutamic acid in the presence of glutamine synthetase to produce ADP; performing a second reaction in which the produced ADP is reacted with glucose in the presence of ADP-dependent hexokinase to produce glucose-6-phosphate; performing a third reaction in which the produced glucose-6-phosphate is reacted with an oxidized NAD compound in the presence of glucose-6-phosphate dehydrogenase to produce a reduced NAD compound; and quantifying the reduced NAD compound to quantify ammonia. 1. A method of quantifying ammonia , the method comprising:performing a first reaction in which a test liquid containing ammonia is reacted with ATP and L-glutamic acid in the presence of glutamine synthetase to produce ADP;performing a second reaction in which the produced ADP is reacted with glucose in the presence of ADP-dependent hexokinase to produce glucose-6-phosphate;performing a third reaction in which the produced glucose-6-phosphate is reacted with an oxidized NAD compound in the presence of glucose-6-phosphate dehydrogenase to produce a reduced NAD compound; andquantifying the reduced NAD compound to quantify ammonia.2. The method of quantifying ammonia according to claim 1 , wherein quantifying the reduced NAD compound to quantify ammonia is carried out by quantifying a pigment obtained by reacting the produced reduced NAD compound with a coloring agent.3. The method of quantifying ammonia according to claim 1 , wherein the first reaction further includes at least one of a magnesium ion (Mg) or a manganese ion (Mn) claim 1 , as a catalyst to produce the ADP.4. The method of quantifying ammonia according to claim 2 , wherein the first reaction further includes at least one of a magnesium ion (Mg) or a manganese ion (Mn) claim 2 , as a catalyst to produce the ADP.5. A quantification reagent kit claim 2 , comprising:a first ...

Apparatus And System For Biofluid Sample Dispensing And/Or Assay

An apparatus for assaying biofluid includes a receptacle for removably housing a set of cuvettes of a biofluid sample dispensing apparatus, each cuvette of the set of cuvettes containing an assay sample having a reagent and a sample of the biofluid; a receptacle sealing element disposed around the receptacle for providing sealing engagement between the receptacle and the biofluid sample dispensing apparatus for sealing the set of cuvettes within the receptacle; a vacuum source for evacuating the receptacle; and an automated assay system connected to the receptacle for performing an assay process on the assay samples in the set of cuvettes, while maintaining the set of cuvettes within the receptacle. 1. An apparatus for assaying biofluid , the apparatus comprising:a receptacle for removably housing a set of cuvettes of a biofluid sample dispensing apparatus, each cuvette of the set of cuvettes containing an assay sample comprising a reagent and a sample of the biofluid;a receptacle sealing element disposed around the receptacle for providing sealing engagement between the receptacle and the biofluid sample dispensing apparatus for sealing the set of cuvettes within the receptacle;a vacuum source for evacuating the receptacle; andan automated assay system connected to the receptacle for performing an assay process on the assay samples in the set of cuvettes, while maintaining the set of cuvettes within the receptacle.2. The apparatus according to claim 1 , wherein the receptacle comprises a set of receptacle sockets claim 1 , each receptacle socket of the set of receptacle sockets for receiving a respective cuvette of the set of cuvettes.3. The apparatus according to claim 2 , wherein the set of receptacle sockets are arranged such that the biofluid sample dispensing apparatus is receivable by the receptacle in only one orientation.4. The apparatus according to claim 2 , wherein the set of receptacle sockets are arranged such that the assay process is performable on ...

Biosensors that detect nad+

A polypeptide biosensor that detects free NAD + is disclosed. The polypeptide comprises a first fragment from an NAD + dependent DNA ligase acetylation domain, a second fragment from the NAD + dependent DNA ligase acetylation domain, and a fluorescent protein, wherein the fluorescent protein is positioned between the two DNA ligase acetylation domain fragments. Also disclosed are expression vectors comprising the biosensor as well as methods of using the biosensor to detect NAD + .

METHODS FOR THE IDENTIFICATION OF BIFUNCTIONAL COMPOUNDS

The present disclosure provides bifunctional compounds including a polypeptide targeting moiety conjugated to a small molecule in a site-specific manner both of which bind to the same target protein resulting in potent and specific binding characteristics and methods of identifying such compounds. 1. A synthetic bifunctional compound , or a pharmaceutically acceptable salt thereof , that modulates the activity of an extracellular target protein , the compound comprising a polypeptide targeting moiety that binds to the extracellular target protein covalently conjugated to a small molecule moiety that binds to the extracellular target protein ,wherein the bifunctional compound binds to the target protein with at least 5-fold greater affinity and/or 5-fold greater selectivity than the affinity of each of the polypeptide targeting moiety and the small molecule moiety alone.2. The synthetic bifunctional compound of claim 1 , wherein the polypeptide targeting moiety is an antibody or an antigen binding fragment thereof or a fibronectin type III domain.3. The synthetic bifunctional compound of claim 2 , wherein the antibody or an antigen binding fragment thereof is an antibody or a fibronectin type III domain.419-. (canceled)20. The synthetic bifunctional compound of claim 1 , wherein the extracellular target protein is a carbonic anhydrase.21. (canceled)23. The synthetic bifunctional compound of claim 1 , wherein the extracellular target protein is a metalloprotease.24. (canceled)26. The synthetic bifunctional compound of claim 1 , wherein the extracellular target protein is PSMA.27. (canceled)29. (canceled)30. The bifunctional compound of claim 1 , wherein the extracellular target protein is CXCR4.31. (canceled)33. (canceled)34. A method of identifying a compound that modulates the activity of a target protein claim 1 , the method comprising:(a) providing two or more synthetic bifunctional compounds comprising a polypeptide targeting moiety covalently conjugated to a ...

STRUCTURE-ACTIVITY RELATIONSHIPS

The present disclosure relates to compositions and methods for screening a plurality of polypeptide variants. 2. The kit of claim 1 , wherein said at least one ligand is immobilized in the wells of said microtiter plate.3. The kit of claim 1 , wherein each of the wells of said microtiter plate contain different ligands.4. A kit for identifying at least one polypeptide having a desired activity for a ligand of interest claim 1 , containing:(i) at least one plurality of active addressable polypeptide variants, wherein each member of each plurality of the active polypeptide variants reacts with at least one ligand to produce a detectable signal, and wherein at least two members of each plurality of the active polypeptide variants are related to a parent polypeptide and react with different ligands, wherein the different ligand does not react with the parent polypeptide wherein each member of said plurality of active polypeptide variants is placed within a well of a microtiter plate;(ii) at least one ligand; and(iii) instructions for use.5. The kit of claim 4 , wherein each member of said plurality of polypeptide variants is immobilized in the wells of said microtiter plate.6. The kit of claim 4 , wherein at least two members of each plurality of active addressable polypeptide variants are related to a parent polypeptide and react with differing levels of activity upon said at least one ligand.7. The kit of claim 4 , further comprising at least one means of generating a signal.8. The kit of claim 8 , further comprising at least one means of detecting said signal.9. The kit of claim 1 , wherein the plurality of active addressable polypeptide variants are operable under conditions in which the parent polypeptide is inactive or rendered inactive.10. The kit of claim 4 , wherein the plurality of active addressable polypeptide variants are operable under conditions in which the parent polypeptide is inactive or rendered inactive.11. The kit of claim 1 , wherein the plurality ...

AGENTS AND METHODS TO ELICIT ANTI-TUMOR IMMUNE RESPONSE

The invention provides an isolated, purified population of human cells comprising CD8 T cells with reduced Cbl-b activity. The invention provides uses of such cells in methods for inducing or enhancing an anti-tumor immune response in a subject. These methods comprise: (a) providing a cell population, from a subject or from another source, which comprises CD8 T cells, (b) reducing Cbl-b activity in the CD8 T-cells, (c) administering the cells of step (b) to the subject. The invention provides methods for making CD8 T cells that do not require stimulation through a co-receptor in order for the cell to become activated or proliferated in response to contact via its T cell receptor. Such methods are based upon reducing function of Cbl-b. The invention also provides methods for identifying agents which affect Cbl-b expression or activity. 118-. (canceled)19. A method for making CD8+ T cells which do not require co-stimulation in order to become activated and proliferate , the method comprising:(a) providing, from peripheral blood of a subject, a cell population comprising CD8+ T cells,(b) reducing Cbl-b activity in the CD8+ T-cells by introducing an siRNA which targets Cbl-b, thereby creating CD8+ T cells which do not require co-stimulation to become activated and proliferate.20. The method of claim 19 , comprising a step of stimulating the CD8+ T cells to proliferate claim 19 , wherein the step is performed after step (b).21. The method of claim 19 , wherein the subject has one or more solid tumors.22. The method of claim 21 , wherein at least one of the one or more solid tumors expresses MHC-I with an antigen that can be recognized by cytotoxic T lymphocytes (CTLs).23. The method of claim 21 , wherein one or more of the solid tumors is a metastatic tumor.24. The method of claim 19 , wherein the siRNA comprises SEQ ID NO: 1 (5′-CAGGAGTATGAGACAGAAG-3′).25. A method for inducing an anti-tumor immune response in a subject in need thereof claim 19 , the method comprising:( ...

ADDING NUCLEOTIDES DURING SEQUENCE DETECTION

Polynucleotide sequencing methods include incubating unlabeled nucleotides with a cluster of template polynucleotide strands having the same sequence when the identity of the previously added labeled nucleotide is being detected. The detection step provides time for the addition of the unlabeled nucleotides to be incorporated into the copy strands in which the previously added labeled nucleotide did not get incorporated. Thus, at the end of the detection step, all or most of the copy strands will be in phase and ready to incorporate the appropriate labeled nucleotide in the subsequence incorporate step. 1. A polynucleotide sequencing method comprising:(a) introducing a first chain extending enzyme and a mixture of blocked, labeled nucleotides to a flow cell comprising a site at which multiple template polynucleotide strands having the same nucleotide sequence are bound to a surface of the flow cell, wherein the first chain extending enzyme is configured to incorporate an appropriate one of the blocked, labeled nucleotides into copy polynucleotide strands, based on the sequence of the template strands to which the copy polynucleotide strands correspond;(b) washing unincorporated blocked, labeled nucleotides away from the flow cell;(c) introducing a composition comprising a mixture of blocked, unlabeled nucleotides to the flow cell during or after the washing away of the unincorporated blocked, labeled nucleotides from the flow cell, wherein a blocked, unlabeled nucleotide of the mixture of the blocked, unlabeled nucleotides is available for incorporation into the copy polynucleotide strands, based on a sequence of the template strands to which the copy polynucleotide strands correspond, provided that the previously incorporated nucleotide in the copy strand, if any, is not blocked; and(d) detecting the identity of the one blocked, labeled nucleotide incorporated into the copy strands, if any, while the mixture of blocked, unlabeled nucleotides is incubated with the ...

ANIMAL MODEL OF AUTISM

Some aspects of this invention provide a non-human animal model of autism. Some aspects of this invention provide a non-human animal model for diseases or disorders associated with an overexpression or a copy number variance of a Ube3a gene. Transgenic mammals and transgenic mammalian cells comprising an exogenous copy or exogenous copies of a ube3a protein-encoding nucleic acid sequence are also provided. Some aspects of this invention further provide methods for using the animal models, cells, and transgenic animals for identifying agents or interventions that can alleviate a pathogenic characteristic observed in the animal model, cell, or transgenic animal. 1. An isolated transgenic mammalian cell comprisingone or more isolated nucleic acid sequence(s) encoding a ubiquitin ligase 3a (ube3a) protein; orone or more exogenous nucleic acid sequence(s) encoding a ube3a protein;one or more recombinant nucleic acid sequence(s) encoding a ube3a protein; orone or more nucleic acid sequence(s) encoding a ube3a protein in addition to any endogenous copies of nucleic acid sequences encoding a ube3a protein.2. The transgenic mammalian cell of claim 1 , wherein the nucleic acid sequence(s) encoding a ube3a protein are stably integrated into the genome of the cell.36-. (canceled)7. The transgenic mammalian cell of claim 1 , wherein the genome of the transgenic mammal comprises three endogenous nucleic acid sequences encoding a ube3a protein claim 1 , optionally wherein the genome of the transgenic mammal comprises an idic15 mutation.8. The transgenic mammalian cell of claim 1 , wherein the cell is a human cell claim 1 , a non-human mammalian cell claim 1 , or a mouse cell.9. The transgenic mammalian cell of claim 7 , wherein the mouse cell is derived from a mouse of FVB claim 7 , dup15 or idic15 genetic background.1011-. (canceled)12. The transgenic mammalian cell of claim 1 , wherein the one or more isolated nucleic acid sequence(s) encoding a ube3a protein comprise a ube3a ...

METHOD FOR SIMULTANEOUSLY MEASURING ACTIVITY OF VARIOUS ENZYMES BY USING MULTI-WAVELENGTH ABSORPTION SINGLE CHANNEL

A method for simultaneously measuring the activities of multiple kinds of enzymes in single channel through concimitantly monitoring multiple wavelength absorbances is claimed; based on linear additivity of absorbances and the isoabsorbance wavelengths of chromogenic substrates and their chromogenic products, both the principles for the combination of chromogenic substrates required for simultaneously measuring the activities of multiple kinds of enzymes in single channel through concimitantly monitoring multiple wavelength absorbances and an approach for data processing to eliminate the interference of the overlapped absorbances are developed, kinetic analysis of reaction curves via numerical integration eliminates the interference of all substrates and products and estimates the maximal reaction rates of the enzymes to be measured, giving the linear ranges and the limits of quantification for simultaneously measuring the activities of multiple kinds of enzymes in single channel comparable to those by separate assays; the present invention is applicable for simultaneously measuring the activities of multiple kinds of enzymes in biological samples, simultaneously measuring multiple components by enzyme-labeled immunoassays, screening simultaneously of inhibitors against multiple targets, for the practice in clinical biochemical analyses, clinical immunoassays, health laboratory analyses, the discovery of inhibitors, and the basical researches which need the present invention. 1. A method for simultaneously measuring the activities of multiple kinds of enzymes in single channel through concomitantly monitoring multiple wavelength absorbances the method comprising:a. determining the combination of chromogenic substrates required for simultaneously measuring the activities of multiple kinds of enzymes:a1. selecting the combination of chromogenic substrates that meets the following criteria according to the specificity of the enzymes to be measured: the selected ...

PROBES AND ASSAYS FOR MEASURING E3 LIGASE ACTIVITY

Provided herein is technology relating to the biological process of protein ubiquitination and particularly, but not exclusively, to compositions and methods for studying protein ubiquitination and developing therapeutics to modulate protein ubiquitination. 120-. (canceled)21. A method of screening for modulators of E3 ligase activity , the method comprising:a) providing a test composition comprising an E3 ligase, a ubiquitin C-terminal thioester fluorophore, and a molecule to test for the ability to modulate the activity of the E3 ligase; andb) monitoring the fluorescence of the fluorophore in the test composition.22. The method of further comprising comparing the fluorescence of the fluorophore in the test composition to the fluorescence of a control composition comprising the E3 ligase and the ubiquitin C-terminal thioester fluorophore claim 21 , but not comprising the molecule to test.23. The method of further comprising identifying the molecule to test as a molecule that modulates the activity of E3 ligase when the fluorescence of the fluorophore in the test composition is different than the fluorescence of the fluorophore in the control composition.24. The method of wherein the E3 ligase is NEDD4 claim 21 , NEDD4L claim 21 , ITCH claim 21 , WWP1 claim 21 , WWP2 claim 21 , SMURF1 claim 21 , SMURF2 claim 21 , NEDL1 claim 21 , NEDL2 claim 21 , E6AP claim 21 , HECTD2 claim 21 , KIAA0614 claim 21 , TRIP12 claim 21 , G2E3 claim 21 , EDD claim 21 , HACE1 claim 21 , HECTD1 claim 21 , UBE3B claim 21 , UBE3C claim 21 , KIAA0317 claim 21 , HUWE 1 claim 21 , HECTD3 claim 21 , HERC1 claim 21 , HERC2 claim 21 , HERC3 claim 21 , HERC4 claim 21 , HERC5 claim 21 , HERC6 claim 21 , SopA claim 21 , NIeL claim 21 , ARIH1 claim 21 , ARIH2 claim 21 , CUL9 claim 21 , ANKIB1 claim 21 , PARK2 claim 21 , RNF144A claim 21 , RNF144B claim 21 , RBCK1 claim 21 , RNF19A claim 21 , RNF19B claim 21 , RNF31 claim 21 , RNF216 claim 21 , RNF14 claim 21 , RNF217 claim 21 , or a NEL E3.25. The ...

METHOD AND ARRAY FOR IDENTIFYING HISTONE-CODE-RELATED ANALYTES

Disclosed embodiments concern an array for use in identifying or identifying and quantifying analytes in a sample using a macrocyclic sensor comprising a macrocyclic compound and a detectable moiety. The disclosed array may be used to discriminate among various analytes based on different features, such as post-translational modifications, isomeric post-translational modifications, and the peptide sequence around post-translational modifications. Also disclosed is a method for identifying analytes comprising a post-translational modification, as well as an enzymatic assay using the disclosed macrocyclic sensor. 2. The macrocyclic compound of claim 1 , wherein:{'sup': '1', 'sub': 1', '8', '1', '8, 'Ris selected from a branched or straight-chain C-C-alkyl or C-Calkenyl; and'}{'sup': 3', 'a', 'a', 'b', 'a', 'b', 'c', '+', 'a', 'b', 'c, 'sub': 2', '2', 'm', '2', '1', '8', '1', '8', '2', '1', '10', '1', '10', '1', '10, 'Ris selected from F, Cl, Br, I, NH, NHR, NRR, or (NRRR), where R, Rand Rindependently are selected from (CH)-alkyl, aryl, or —SOaryl, where m=0, 1, or 2 and alkyl is a branched, straight-chain or cyclic C-Calkyl or C-Calkenyl, and the —SOaryl group is optionally substituted with one or more functional groups selected from cyano, amide, carboxyl, carboxylic acid, amine, alkyl amine, C-Calkyl, C-Calkoxy, halo, aldehyde, hydroxyl, C-Calkylhydroxyl, or combinations thereof.'}3. The macrocyclic compound of claim 1 , wherein Ris heteroaryl claim 1 , aryl or heteroaliphatic claim 1 , and is optionally substituted with one or more functional groups selected from cyano claim 1 , amide claim 1 , carboxyl claim 1 , carboxylic acid claim 1 , amine claim 1 , alkyl amine claim 1 , C-Calkyl claim 1 , C-Calkoxy claim 1 , halo claim 1 , aldehyde claim 1 , hydroxyl claim 1 , C-Calkylhydroxyl claim 1 , or combinations thereof.4. An array claim 1 , comprising at least two macrocyclic sensors capable of identifying claim 1 , or identifying and quantifying claim 1 , an analyte ...

Method for Identifying Polyubiquitinated Substrate

An object of the present invention is to provide a method for efficiently identifying a polyubiquitinated substrate which is generally not easily identified. The method for identifying a polyubiquitinated substrate includes (1) a step of expressing a trypsin-resistant polyubiquitin chain-binding protein and a ubiquitin ligase in a cell, (2) a step of isolating a complex that contains the trypsin-resistant polyubiquitin chain-binding protein from the cell having undergone the step (1), (3) a step of subjecting the complex isolated by the step (2) to trypsin digestion, and (4) a step of identifying a peptide that has a ubiquitination site from a digested material obtained by the step (3). 1. A method for identifying a polyubiquitinated substrate , comprising:(1) a step of expressing a trypsin-resistant polyubiquitin chain-binding protein and a ubiquitin ligase in a cell or a cell lysate;(2) a step of isolating a complex that contains the trypsin-resistant polyubiquitin chain-binding protein from the cell or the cell lysate having undergone the step (1);(3) a step of subjecting the complex isolated by the step (2) to trypsin digestion; and(4) a step of identifying a peptide that has a ubiquitination site from a digested material obtained by the step (3).2. The method for identifying a polyubiquitinated substrate according to claim 1 , further comprising:(1′) a step of expressing the trypsin-resistant polyubiquitin chain-binding protein and a dominant-negative mutant of the ubiquitin ligase in another cell or another cell lysate of the same kind as the aforementioned cell;(2′) a step of isolating a complex that contains the trypsin-resistant polyubiquitin chain-binding protein from the cell or the cell lysate having undergone the step (1′);(3′) a step of subjecting the complex isolated by the step (2′) to trypsin digestion;(4′) a step of identifying a peptide that has a ubiquitination site from the digested material obtained by the step (3′); and(5) a step of ...

COMPOSITIONS AND METHODS RELATING TO ARGININOSUCCINATE SYNTHETASE

Processes and compositions for the therapeutic treatment of pathogenic Gram-negative bacterial infection are provided whereby argininosuccinate synthetase or PEGylated argininosuccinate synthetase is administered to a subject to inactivate endotoxin thereby reducing the likelihood of bacterial sepsis and improving patient outcome. 1. A process of detecting exposure to bacterial endotoxin by a subject , comprising:obtaining a sample from said subject exposed to a bacterial endotoxin absorbed from an infected wound and circulating in blood of said subject prior to said obtaining;determining that levels and activity of argininosuccinate synthetase in said sample from said subject are elevated over a normal level and normal activity of arginosuccinate synthetase absent exposure to bacterial endotoxin.2. The process of wherein said sample comprises serum.3. The process of wherein said subject is infected with a pathogenic Gram-negative bacteria as a source of said bacterial endotoxin.4. The process of wherein the argininosuccinate synthetase has an amino acid sequence of SEQ ID NO: 1.5. The process of wherein said argininosuccinate synthetase protein has an amino acid sequence of a truncation of SEQ ID NO: 1 retaining a binding site for said bacterial endotoxin.6. The process of wherein said subject is human. This application is a divisional application of U.S. Utility application Ser. No. 14,116,817 filed Nov. 11, 2013 that in turn is a US National Phase Application of PCT Application PCT/US2011/022561 filed Jan. 26, 2011 that in turn claims priority to U.S. Provisional Application No. 61/298,309 filed Jan. 26, 2010, the entire contents of which are incorporated herein by reference.The invention relates generally to compositions and methods for detection and treatment of microbial infection in a subject. In specific embodiments, the present invention relates to compositions and methods for detection and treatment of exposure to bacterial endotoxin in a subject.Infection ...

METHODS FOR SCREENING UBIQUITIN LIGASE AGONISTS

Disclosed herein are methods for identifying a ubiquitin ligase agonist, and the methods include (a) contacting a ubiquitin ligase with a candidate agonist and a neo-substrate; and (b) determining whether the candidate agonist is effective to result in binding the ubiquitin ligase to the neo-substrate, wherein binding of the ubiquitin substrate to the neo-substrate identifies the candidate agonist as a ubiquitin ligase agonist. 1. A method for identifying a ubiquitin ligase agonist , comprising:(a) contacting a ubiquitin ligase with a candidate agonist and a neo-substrate; and(b) determining whether the candidate agonist is effective to result in binding the ubiquitin ligase to the neo-substrate, wherein binding of the ubiquitin substrate to the neo-substrate identifies the candidate agonist as a ubiquitin ligase agonist.2. The method of claim 1 , wherein binding of the ubiquitin ligase to the neo-substrate provides a complex comprising the ubiquitin ligase claim 1 , the agonist claim 1 , and the neo-substrate.3. The method of claim 1 , wherein determining whether the candidate agonist is effective to result in binding the ubiquitin ligase to the neo-substrate comprises observing a signal generated by the binding.4. The method of claim 1 , wherein the ubiquitin ligase further comprises a first reporting agent and the substrate further comprises a second reporting agent claim 1 , wherein upon binding of the ubiquitin ligase to the neo-substrate claim 1 , the first reporting agent acts with the second reporting agent to generate a signal.5. The method of claim 4 , wherein the ubiquitin ligase comprising the first reporting agent is a donor bead configured to generate a reactive oxygen species when in the excited state claim 4 , and the neo-substrate comprising the second reporting agent is an acceptor bead configured to generate light in the presence of a reactive oxygen species upon binding of the ubiquitin ligase to the neo-substrate.6. The method of claim 5 , ...

Methods For Detecting Or Quantifying CTP And CTP Synthase Activity

The present invention relates to a method for detecting or quantifying CTP in a cell sample comprising at least two nucleotide triphosphates by cationic ion pairing chromatography coupled to mass spectrometry, to a method for detecting or quantifying CTP synthase activity based on the method for detecting or quantifying CTP, and to their use in methods for screening potential immunosuppressive or anti-cancer compounds and in methods for determining the appropriate dose of an immunosuppressive or anti-cancer compound inhibiting CTP synthase activity for a treated subject. 1. A method for detecting or quantifying CTP in a sample comprising at least two distinct nucleotide triphosphate , said method comprising:a) separating the distinct nucleotide triphosphate by ion-pairing chromatography using a cationic ion pair reagent contained in a mobile phase having a pH comprised between 6 and 7, wherein the sample injected for the chromatography has a pH comprised between 1 and 7 andb) detecting or quantifying CTP by mass spectrometry.2. The method according to any one of claim 1 , wherein step a) is performed by using a reverse phase column claim 1 , preferably comprising beads of a diameter inferior or equal to 10 μm grafted with a hydrophobic molecule.3. The method according to claim 2 , wherein the column is High Strength Silica (HSS) column.4. The method according to any one of to claim 2 , wherein the cationic ion pair reagent used in step a) is dibutylamine acetate or tributylamine acetate.5. The method of any one of to claim 2 , wherein the concentration of the cationic ion pair reagent in step a) is comprised between 5 and 10 mM claim 2 , preferably between 5 and 8 mM and more preferably claim 2 , between 5 and 6 mM.6. The method of any one of to claim 2 , wherein a stable isotope CTP standard is added to the sample before step a).7. The method according to any one of to claim 2 , wherein the separation in step a) is obtained by forming a binary gradient of an ...

HIGH-THROUGHPUT ABSORBANCE MEASUREMENTS OF SAMPLES IN MICROCAPILLARY ARRAYS

The present invention provides a method for measuring the amount of absorbance of a sample in a microcapillary based on measuring the absorbance in the sample. 1. A high-throughput method for determining the absorbance for multiple samples in a microcavity array , the method comprising:i) transmitting light of a definable wavelength through samples contained in said microcavity array, wherein one sample is loaded into each microcavity within the array;ii) measuring the light transmitted through said samples with a detector, wherein the light transmitted is measured for each individual sample within the array in order to obtain a light transmittance intensity for each individual sample within the array;iii) comparing the light transmittance intensity obtained for each individual sample in step ii) to the light transmittance intensity for a control sample; andiv) calculating the absorbance of each individual sample in the array based on the comparison in step iii) in order to determine spectrometric differences between said samples.6. The method of claim 2 , wherein the absorbance is calculated by the following formula:{'br': None, 'i': A', 'T., 'sub': '10', '=−log'}7. The method of claim 3 , wherein the absorbance is calculated by the following formula:{'br': None, 'i': A=', 'T, 'sub': '10', '2−log%.'}8. The method of claim 1 , wherein said method further comprises using said absorbance to determine one or more spectrometric characteristics.9. The method of claim 8 , wherein said spectrometric characteristics are selected from the group consisting of concentration claim 8 , enzyme activity claim 8 , enzyme-substrate interaction claim 8 , receptor-ligand binding claim 8 , affinity binding claim 8 , stability claim 8 , and cell growth.10. The method of claim 1 , wherein said enzyme activity results are based on a colorimetric assay.11. The method of claim 1 , wherein said concentration or cell growth results are based on a densitometric assay.12. The method of claim 10 ...

Tobacco Smoke Detector

A disposable, self-contained tobacco smoke detector, comprising a sensor incorporated with a reagent capable of undergoing one or more chemical reactions by interacting with one or more compounds unique to tobacco smoke. The smoking detector further comprises an indicator coupled to the sensor for visually indicating the occurrence of at least one chemical reaction by producing a detectable change in color, indicating exposure to tobacco smoke, thereby providing visual evidence of smoking. 1. A disposable , self-contained , tobacco smoke detector , comprising:a sensor incorporated with a reagent capable of undergoing at least one chemical reaction by interacting with one or more compounds unique to tobacco smoke; andan indicator coupled to the sensor for visually indicating the occurrence of the at least one chemical reaction by producing a detectable change in color so as to form a detectable color change; andwherein when the reagent interacts with the one or more compounds unique to the tobacco smoke, the at least one chemical reaction formed thereby has the detectable color change made thereto, with the detectable color change being visible via the indicator so as to provide a visual indication of the tobacco smoke.2. The tobacco smoke detector of claim 1 , further comprises a housing adapted to contain the sensor and the indicator.3. The tobacco smoke detector of claim 1 , wherein the reagent comprises at least an enzyme or a protein or a chemical substance.4. The tobacco smoke detector of claim 1 , wherein the sensor is integrated with the indicator.5. The tobacco smoke detector of claim 1 , is a self-contained unit allowing for easy transport claim 1 , installation claim 1 , and replacement.6. The tobacco smoke detector of claim 1 , is tamper resistant.7. The tobacco smoke detector of claim 1 , wherein the biological or chemical component is capable of initiating a series of chemical reactions in the presence of tobacco smoke or compounds unique to tobacco ...

Adenylation Enzyme Inhibitors

The present invention concerns compounds that are capable of covalently entrapping adenylating enzymes. The present invention is essentially based on the discovery that analogues of adenylating enzyme (AE) substrates, wherein a methylene group has been incorporated at the carbon atom in the α-position relative to the carboxylate group involved in the adenylation, are capable of undergoing the adenylation reaction, thereby creating an activated methylene group in situ. The resulting ‘armed’ acyladenylate can interact with the enzyme resulting in covalent entrapment. Interestingly, the acyladenylate can alternatively be transferred to the next step in the enzyme cascade, following which the activated methylene group can interact with the next (active site cysteine containing) enzyme in the enzymatic cascade. The AE substrate analogues, based on their capability of ‘entrapping’ the respective enzymes, will have utility as activity based probes in biological research and also as diagnostic and/or therapeutic agents. 3. The adenylating enzyme (AE) substrate analogue according to claim 1 , wherein the capability of the compound binding to an adenylating enzyme (AE) is established using a competition binding assay claim 1 , wherein an ICvalue of less than 10 μM is indicative of said capability of binding to an adenylating enzyme (AE).5. The adenylating enzyme (AE) substrate analogue according to claim 4 , wherein the adenylating enzyme is an E1 enzyme involved in the ubiquitin conjugation pathway claim 4 , an E1 enzyme involved in the SUMO conjugation pathway claim 4 , an E1 enzyme involved in the NEDD8 conjugation pathway claim 4 , or an enzyme involved in the ISG15 pathway.6. The adenylating enzyme (AE) substrate analogue according to claim 4 , wherein X represents —NH— and R represents a peptide selected from the group consisting of Ub(1-75) claim 4 , NEDD8(1-75) claim 4 , ISG15(1-156) claim 4 , SUMO1(1-96) claim 4 , SUMO2(1-92) claim 4 , SUMO3(1-91) claim 4 , UFM1(1-82 ...

BIOMARKERS FOR RAPID DETECTION OF AN OCCURRENCE OF A STROKE EVENT

A diagnostic assay for detecting an occurrence of a stroke event in a mammalian subject. The assay comprises the steps of: (i) separating a plasma fraction from a blood sample collected from the mammalian subject; (ii) quantifying in the plasma fraction a L-glutamine hydroxylamine glutamyl transferase (L-GHGT) activity; (iii) quantifying in the plasma fraction a gamma glutamyl hydroxamate synthetase (GGHS) activity; (iv) adding together or alternatively calculating the combinatorial probability for the quantified L-GHGT activity and the quantified GGHS activity to produce a value for the net glutamine synthetase activity, and (v) correlating the net glutamine synthetase activity value with net glutamine synthetase activity values from healthy subjects to detect an occurrence of a stroke event in the mammalian subject. Also disclosed are kits comprising reagents and instructions for performing a diagnostic assay to detect and quantify L-GHGT activity and/or GGHS activity. 1. A diagnostic assay for detecting an occurrence of a stroke event in a mammalian subject , comprising:separating a plasma fraction from a blood sample collected from the mammalian subject;quantifying in the plasma fraction a L-glutamine hydroxylamine glutamyl transferase (L-GHGT) activity;quantifying in the plasma fraction a gamma glutamyl hydroxamate synthetase (GGHS) activity;producing a value for glutamine synthetase activity by adding together the quantified L-GHGT activity and the quantified GGHS activity or alternatively, by estimating combinatorial probabilities of the quantified L-GHGT activity and the quantified GGHS activity; andcorrelating said glutamine synthetase activity value with a glutamine synthetase activity value from a healthy subject to detect an occurrence of a stroke event.2. The diagnostic assay of claim 1 , wherein the L-GHGT activity is detected with an assay comprising three sets of steps wherein: [{'sub': 2', '2, 'mixing together (i) a pH 6.5 imidazole buffer, (ii) a ...

Use of a Reverse-Transcriptase Inhibitor in the Prevention and Treatment of Degenerative Diseases

The invention relates to the use of a reverse-transcriptase inhibitor in the prevention or treatment of a degenerative disease. 1. A method of treating or preventing a degenerative disease in a subject in need thereof , the method comprising administering a therapeutically effective amount of a reverse transcriptase inhibitor to the subject.2. The method of claim 1 , wherein the reverse transcriptase inhibitor is a nucleoside inhibitor.3. The method of claim 2 , wherein the degenerative disease is linked to aging or oxidative stress.4. The method of claim 3 , wherein the degenerative disease is a neurodegenerative disease.5. The method of claim 4 , wherein the degenerative disease is selected from the group consisting of Parkinson's disease claim 4 , Alzheimer's disease claim 4 , Huntington's disease claim 4 , amyotrophic lateral sclerosis (ALS) claim 4 , degenerative diseases affecting eyesight and degenerative diseases affecting hearing.6. The method of claim 5 , wherein the degenerative disease is glaucoma. The present invention relates to the use of a reverse transcriptase inhibitor in the prevention and treatment of degenerative diseases.A characteristic of most degenerative diseases, in particular neurodegenerative diseases, is that they manifest late in life. This is true of both their sporadic and genetic forms, as is illustrated by Parkinson's disease, Alzheimer's disease and even the monogenetic Huntington's disease. This suggests that aging makes the cells more sensitive, even if the mutations themselves accelerating this aging cannot be ruled out.Oxidative stress, which mimics accelerated aging, generates reactive oxygen species (ROS) which are toxic, in particular in terms of the genome, where the chromatin proteins and the DNA bases are subjected to oxidation (Vijg, J., and Suh, Y., 2013). Reactive oxygen species may induce single-stranded and double-stranded (DSB) DNA breaks, thereby activating DNA damage response systems (O'Sullivan and Karlseder, ...

ARTERIOSCLEROSIS AND CANCER DETECTION METHOD USING DEOXYHYPUSINE SYNTHASE GENE AS INDICATOR

The present inventors have surprisingly found that a deoxyhypusine synthase (DHPS) gene, which was previously reported to correlate with prostate cancer and cervical cancer, is highly responsive to arteriosclerosis or digestive system cancer, whereby the gene can be used as a desired marker for arteriosclerosis or digestive system cancer. The present invention has been accomplished on the basis of this finding. Specifically, the present invention provides a method for determining arteriosclerosis or digestive system cancer, which method includes detecting expression of a deoxyhypusine synthase gene in a test sample (preferably a blood sample), and determining arteriosclerosis or digestive system cancer of a test subject from which the test sample has been obtained, on the basis of an increase in the gene expression as an index. 1. A method for determining arteriosclerosis , which method comprises detecting expression of a deoxyhypusine synthase gene in a test sample , and determining arteriosclerosis of a test subject from which the test sample has been obtained , on the basis of an increase in the gene expression as an index.2. The arteriosclerosis determination method according to claim 1 , wherein expression of the deoxyhypusine synthase gene claim 1 , which is a target gene claim 1 , is confirmed by detecting the entirety or a part of the deoxyhypusine synthase encoded by the gene claim 1 , as a target polypeptide.3. The arteriosclerosis determination method according to claim 2 , wherein the deoxyhypusine synthase claim 2 , which is a target polypeptide claim 2 , is detected by use of an antibody which specifically binds to the target polypeptide.4. The arteriosclerosis determination method according to claim 3 , wherein the antibody is a monoclonal antibody.5. The arteriosclerosis determination method according to claim 1 , wherein expression of the deoxyhypusine synthase gene claim 1 , which is a target gene claim 1 , is confirmed by detecting an antibody ...

DEVICES HAVING A SAMPLE DELIVERY COMPONENT

Examples herein provide a device. The device includes a sample delivery component, which includes: a reagent chamber to contain at least one reagent; a sample chamber to contain a fluid sample; and a delivery channel extending from the reagent chamber and in fluid communication with the sample chamber and an output port, wherein the delivery channel is conducive to mixing the at least one reagent and the fluid sample to form a mixture before the mixture reaches the output port and be discharged therefrom. The device includes a testing cassette detachable from the delivery component, which includes: an input port in fluid communication with a microfluidic reservoir, the input port to receive the discharged fluid sample from the output port; and a micro-fabricated integrated sensor in a microfluidic channel extending from the microfluidic reservoir. 1. A device , comprising: a reagent chamber to contain at least one reagent;', 'a sample chamber to contain a fluid sample; and', 'a delivery channel extending from the reagent chamber and in fluid communication with the sample chamber and an output port, wherein the delivery channel is conducive to mixing the at least one reagent and the fluid sample to form a mixture before the mixture reaches the output port and be discharged therefrom; and, 'a sample delivery component, comprising an input port in fluid communication with a microfluidic reservoir, the input port to receive the discharged fluid sample from the output port; and', 'a micro-fabricated integrated sensor in a microfluidic channel extending from the microfluidic reservoir., 'a testing cassette detachable from the delivery component, comprising2. The device of claim 1 , wherein the micro-fabricated integrated sensor comprises an impedance-based microchip.3. The device of claim 1 , wherein the device further comprises a transport mechanism to move the fluid sample through the delivery channel by at least one of a capillary pump claim 1 , a thermal inkjet pump ...

Agents and methods for spectrometric analysis

Disclosed herein are agents, methods, and kits for determining the presence or concentration of a target, or multiple targets, in a sample, in a uniplexed or multiplexed fashion. In general, the methods enable the analysis of small molecules produced or consumed in liquid-phase that may be analyzed using gas or vapor phase detection methods.

Аналитическая полоска и способ ее изготовления

1. Аналитическая полоска, которая содержит подложку и канал, отличающаяся тем, что:подложка имеет плоскую поверхность;канал сформирован на плоской поверхности в соответствии с заданной схемой таким образом, что поверхность канала является не ниже, чем плоская поверхность подложки, и канал имеет структуру матрицы, содержащей полости, при этом канал является более гидрофильным, чем плоская поверхность подложки; иреагирующий материал введен в структуру матрицы, содержащей полости.2. Аналитическая полоска по п.1, отличающаяся тем, что поверхность канала выше, чем плоская поверхность подложки.3. Аналитическая полоска по п.2, отличающаяся тем, что канал возвышается в отношении своего сечения над плоской поверхностью.4. Аналитическая полоска по п.1, отличающаяся тем, что канал выполнен из материала, выбранного из группы, состоящей из нитроцеллюлозы и стекловолокна.5. Аналитическая полоска по п.1, отличающаяся тем, что канал сформирован на плоской поверхности в соответствии со способом, выбранным из группы, состоящей из литографской печати, глубокой печати, анастатической печати, трафаретной печати, маркирования линий, струйной печати или нанесения покрытий окунанием.6. Аналитическая полоска по п.1, отличающаяся тем, что канал дополнительно содержит некоторое количество каналов ответвления.7. Аналитическая полоска по п.6, отличающаяся тем, что по меньшей мере один из указанных каналов ответвления дополнительно содержит увеличенную зону, выполненную в форме, выбранной из группы, состоящей из купола, продолговатой формы или формы в виде «острова».8. Аналитическая полоска по п.1, отличающаяся тем, что канал изготовлен из рас� РОССИЙСКАЯ ФЕДЕРАЦИЯ (19) RU (11) (51) МПК G01N 33/53 (13) 2011 141 298 A (2006.01) ФЕДЕРАЛЬНАЯ СЛУЖБА ПО ИНТЕЛЛЕКТУАЛЬНОЙ СОБСТВЕННОСТИ (12) ЗАЯВКА НА ИЗОБРЕТЕНИЕ (21)(22) Заявка: 2011141298/15, 23.03.2009 (71) Заявитель(и): АКТЕРМ ИНК. (CN) Приоритет(ы): (22) Дата подачи заявки: 23.03.2009 (43) Дата публикации заявки: 27.04.2013 Бюл. № 12 R U (85) Дата ...

一种基于共价连接的已知分子与蛋白质相互作用检测系统及其鉴定或验证方法

本发明公开了一种基于共价连接的已知分子与蛋白质相互作用检测系统及其鉴定或验证方法，所述检测系统包括：a)链霉亲和素‑短肽四聚体；b)PafA酶；c)生物素修饰的已知分子。本发明采用生物素标记已知分子，已知分子与蛋白质发生相互作用后，通过链霉亲和素‑短肽四聚体在温和的条件下高效地捕获已知分子的相互作用蛋白质，再在PafA酶的催化下，将短肽与已知分子的相互作蛋白质共价连接，从而将相互作用的已知分子与蛋白质之间的非共价结合转换为链霉亲和素与蛋白质之间的共价连接，从而进行分析，分离和鉴定。本方法可以在保持已知分子天然结构的基础上，实现弱相互作用和瞬时相互作用的捕获，可以用于已知分子与相互作用蛋白质的验证和发现。

Electrochemical biosensor

본 발명은 바이오센서에서 적혈구용적률(hematocrit)의 양에 따른 측정오차를 획기적으로 감소할 수 있는 감응막 조성물 및 이를 포함한 바이오 센서에 관한 것으로서, 구체적으로 효소, 전자전달매개체, 수용성 고분자, 지방산 및 4차 암모늄 염을 포함한 감응막 조성물; 및 상기 조성물로 이루어진 감응막과 혈액시료를 전처리 과정 없이 빠르고 정확하게 도입할 수 있는 시료도입부를 구비한 바이오센서에 관한 것이다. 본 발명의 바이오 센서는 감응막 조성물을 포함하므로서, 바이온센서에서 적혈구용적률의 양에 따른 측정오차를 감소시킬 수 있으며, 새로운 시료도입부를 구비하므로서 수 마이크로 리터 이내의 소량시료를 사용하는 다양한 형태의 전기화학 바이오센서에 쉽게 적용할 수 있다. The present invention relates to a sensitized membrane composition capable of dramatically reducing the measurement error according to the amount of hematocrit in a biosensor, and a biosensor including the same, and specifically, an enzyme, an electron transfer medium, a water-soluble polymer, a fatty acid, and 4 Sensitizing membrane compositions comprising secondary ammonium salts; And it relates to a biosensor having a sample introduction portion that can be introduced quickly and accurately to the sensitized membrane and blood sample consisting of the composition. The biosensor of the present invention includes a sensitizing membrane composition, which can reduce the measurement error according to the amount of red blood cell volume in the biosensor, and has various types of samples using a small sample within several microliters with a new sample introduction unit. Easily applicable to electrochemical biosensors. 바이오센서, 적혈구용적률, 감응막 조성물, 시료도입부, 여분공간부. Biosensor, red blood cell volume, sensitized membrane composition, sample introduction part, extra space part.

For the systems, devices and methods of cell capture and its manufacturing method

本公开内容的实施方案涉及用于评价单个细胞分泌特性谱的系统、方法和设备。在一些实施方案中，所述设备可以被配置成用于分析由生物细胞所表达的物质，并且可以包含第一可压缩基底和被配置成用于与所述第一基底可移除式密封连接的第二基底。在一些实施方案中，在所述第二基底与所述第一基底相连接后，形成组装件，从而使得所述多个腔室的开放侧被所述第二基底覆盖，并且所述多个捕获区域中的每一个的一部分暴露在所述腔室中的每一个之中。

Pluripotent stem cell culture on micro-carriers

본 발명은 마이크로-캐리어 상에서의 만능 줄기 세포의 성장, 증식 및 분화 방법에 관한 것이다. The present invention relates to a method for the growth, proliferation and differentiation of pluripotent stem cells on a micro-carrier.

Method of personalized treatment of chronic renal disease in patients with diabetic nephropathy

FIELD: medicine.SUBSTANCE: present invention relates to a method for personalized treatment of chronic renal disease in patients with diabetic nephropathy, according to which personalization of drug selection, mode and duration of monotherapy or preparations for combined therapy and adding epigallocatechin-3-gallate as an antioxidant depending on the detected urine proteomic profile, intermolecular interactions of the detected peptides and urine proteins.EFFECT: higher clinical effectiveness in chronic renal disease accompanying diabetic nephropathy.1 cl, 2 ex РОССИЙСКАЯ ФЕДЕРАЦИЯ (19) RU (11) (13) 2 687 256 C1 (51) МПК G01N 33/68 (2006.01) C12Q 1/25 (2006.01) A61K 38/17 (2006.01) C12Q 1/68 (2006.01) ФЕДЕРАЛЬНАЯ СЛУЖБА ПО ИНТЕЛЛЕКТУАЛЬНОЙ СОБСТВЕННОСТИ (12) ОПИСАНИЕ ИЗОБРЕТЕНИЯ К ПАТЕНТУ (52) СПК G01N 33/68 (2018.08); C12Q 1/25 (2018.08); A61K 38/17 (2018.08); C12Q 1/68 (2018.08) (21) (22) Заявка: 2018110771, 26.03.2018 (24) Дата начала отсчета срока действия патента: 26.03.2018 08.05.2019 Приоритет(ы): (22) Дата подачи заявки: 26.03.2018 (45) Опубликовано: 08.05.2019 Бюл. № 13 Адрес для переписки: 367000, РД, г. Махачкала, пл. Ленина, 1, Патентный отдел Даггосмедуниверситета (56) Список документов, цитированных в отчете о поиске: Nokiro Yamabe et al: C 1 2 6 8 7 2 5 6 R U (54) Способ персонализированного лечения хронической болезни почек у больных с диабетической нефропатией (57) Реферат: Настоящее изобретение относится к способу эпигаллокатехин-3-галлата в качестве персонализированного лечения хронической антиоксиданта в зависимости от обнаруженного болезни почек у больных с диабетической протеомного профиля мочи, межмолекулярных нефропатией, согласно которому проводят взаимодействий выявленных пептидов и белков персонализацию выбора препарата, режима и мочи. Технический результат – повышение длительности монотерапии или препаратов для эффективности лечения хронической болезни комбинированной терапии и добавление почек при диабетической нефропатии. 2 пр. Стр.: 1 C ...

Fluid test chip and method to make it

一种流体检测试片及其制造方法。该流体检测试片主要包括基板及流道结构，基板具有平面状的第一表面，且流道结构以预设图案形成于基板的第一表面上，使得流道结构的表面高度不低于基板的第一表面的高度。流道结构具有中空网状构造，且流道结构的亲水性高于基板第一表面的亲水性。

Culture of pluripotent stem cells on microcarriers

Изобретение относится к биотехнологии. Раскрыт способ выращивания эмбриональных стволовых клеток человека. Способ предусматривает прикрепление популяции клеток к микроносителю в среде, содержащей ингибитор Rho-киназы. Далее проводят культивирование клеток, отделение клеток от микроносителя и прикрепление полученных клеток ко второму микроносителю в среде, содержащей ингибитор Rho-киназы. Изобретение может быть использовано в медицине. 6 з.п. ф-лы, 48 ил., 3 табл., 11 пр. РОССИЙСКАЯ ФЕДЕРАЦИЯ (19) RU (11) (13) 2 555 538 C2 (51) МПК C12N 5/0735 (2010.01) ФЕДЕРАЛЬНАЯ СЛУЖБА ПО ИНТЕЛЛЕКТУАЛЬНОЙ СОБСТВЕННОСТИ (12) ОПИСАНИЕ (21)(22) Заявка: ИЗОБРЕТЕНИЯ К ПАТЕНТУ 2011124900/10, 19.11.2009 (24) Дата начала отсчета срока действия патента: 19.11.2009 (72) Автор(ы): НЕЛЬСОН, Шелли (US) (73) Патентообладатель(и): СЕНТОКОР ОРТО БАЙОТЕК ИНК. (US) Приоритет(ы): (30) Конвенционный приоритет: (43) Дата публикации заявки: 27.12.2012 Бюл. № 36 R U 20.11.2008 US 61/116,447 (45) Опубликовано: 10.07.2015 Бюл. № 19 2008102118 A1, 28.08.2008. RU 2323252 C1, 27.04.2008 (85) Дата начала рассмотрения заявки PCT на национальной фазе: 20.06.2011 (86) Заявка PCT: 2 5 5 5 5 3 8 (56) Список документов, цитированных в отчете о поиске: WO 2008004990 A2, 10.01.2008. WO 2 5 5 5 5 3 8 R U (87) Публикация заявки PCT: WO 2010/059775 (27.05.2010) Адрес для переписки: 129090, Москва, ул. Б. Спасская, 25, строение 3, ООО "Юридическая фирма Городисский и Партнеры" (54) КУЛЬТУРА ПЛЮРИПОТЕНТНЫХ СТВОЛОВЫХ КЛЕТОК НА МИКРОНОСИТЕЛЯХ (57) Реферат: Изобретение относится к биотехнологии. клеток, отделение клеток от микроносителя и Раскрыт способ выращивания эмбриональных прикрепление полученных клеток ко второму стволовых клеток человека. Способ микроносителю в среде, содержащей ингибитор предусматривает прикрепление популяции клеток Rho-киназы. Изобретение может быть к микроносителю в среде, содержащей ингибитор использовано в медицине. 6 з.п. ф-лы, 48 ил., 3 Rho-киназы. Далее проводят культивирование табл., 11 пр. ...

Methods and compositions related to the modulation of riboswitches

Disclosed herein are methods and compositions related to the detection of conformational changes and interactions with trigger molecules in riboswitches.

Yeast cell surface display technology-based explosive visualization biosensor and preparation method and application thereof

本发明提供了一种基于酵母细胞表面展示技术的爆炸物可视化生物传感器及其制备方法和应用。所述爆炸物可视化生物传感器中含有锚定蛋白的编码基因和爆炸物可视化的目标基因 dntA 或 dntB 基因。本发明利用锚定蛋白将目标基因表达产物锚定到宿主菌表面，最终获得爆炸物可视化生物传感器。该生物传感器能够特异性的将2,4‑二硝基甲苯（2,4‑DNT）降解成一种肉眼可见的红色物质2‑羟基‑5‑甲基苯醌，进而判断待测样品中是否存在爆炸物分子，并且其在420nm处具有最大吸光值，可根据标准曲线计算2,4‑DNT浓度，其稳定性很好，且检测范围宽、灵敏度高、方法简便、成本低、安全性强，因此具有广泛的应用前景。

Reagents for the detection of protein phosphorylation in T-cell receptor signaling pathways

The invention discloses 95 novel phosphorylation sites identified in signal transduction proteins and pathways downstream of the T-cell receptor, and provides phosphorylation-site specific antibodies and heavy-isotope labeled peptides (AQUA peptides) for the selective detection and quantification of these phosphorylated sites/proteins, as well as methods of using the reagents for such purpose. Among the phosphorylation sites identified are sites occurring in the following protein types: Actin Binding proteins, Adaptor/Scaffold proteins, Adhesion proteins, Calcium-binding proteins, Cell Cycle Regulation or Channel proteins, Chaperones, Cofactor proteins, Cytoskeletal proteins, DNA Binding proteins, G protein or GTPase Activating proteins, Ligases, Lipid Kinases and Binding proteins, Oxidoreductases, Protein Kinases, Protein Phosphatases, Receptor proteins, RNA Binding proteins, Transcription Factor/Initiation Complex proteins, Transcription Coactivator/Corepressor proteins, Translation Initiation Complex proteins, Ubitquitin Conjugating System proteins, and Vesicle proteins.

High-throughput assay

A homogeneous high throughput assay is described which screens compounds for enzyme inhibition, or receptor or other target binding. Inhibition (or binding) by the library compounds causes a change in the amount of an optically detectable label that is bound to suspendable cells or solid supports. The amounts of label bound to individual cells or solid supports are microscopically determined, and compared with the amount of label that is not bound to individual cells or solid supports. The degree of inhibition or binding is determined using this data. Confocal microscopy, and subsequent data analysis, allow the assay to be carried out without any separation step, and provide for high throughput screening of very small assay volumes using very small amounts of test compound.

Methods to overcome genetic resistance, improve therapeutic efficacy and minimize safety risks associated with kinase inhibitor therapy

【課題】キナーゼ阻害剤の薬物結合相互作用に基づいて、癌及び他の疾患である患者の治療計画を開発する方法、ならびにキナーゼの変異状態及びキナーゼ占有率を特定する方法を提供する。【解決手段】試験化合物の標的キナーゼ占有率を特定する方法であって、試験試料中の、活性状態にあるが試験化合物と結合していない標的キナーゼの残存レベルを、参照化合物と結合した代表的なシグネチャーペプチドのレベルに基づいて得ることと、なお、試験試料中の活性状態にある標的キナーゼのレベルは、結合型標的キナーゼのレベルと残存レベルとを合算したレベルであり、試験試料中の標的キナーゼの総レベルは、活性状態にある標的キナーゼのレベルと遊離型標的キナーゼのレベルとを合算したレベルである、標的キナーゼ占有率を得ることと、標的キナーゼ占有率は、標的キナーゼの総レベルに対する結合型標的キナーゼのレベルである、方法である。【選択図】なし

Method for the coupled enzyme immunochemical assay of analytes by means of endogenous calibrators

本发明涉及通过分析元件对生物样品和其它液体样品中的多种待分析物(例如代谢物和抗原)进行测定的方法，所述分析元件特别是侧向流动测试带、流通膜体系(流通测试)、微量滴定板或试管的孔/洞，本发明的方法以偶联的酶和亲和反应为基础，并借助于内源校准物(即内源生产的物质)完成，借助于所述内源校准物(例如肌酸酐、葡萄糖、葡萄糖-6-磷酸盐、乳酸盐、谷氨酸盐、天冬氨酸盐、胆固醇、丙酮酸盐、尿素和甘油三酯)能够校正样品基质的稀释度。本发明的应用领域主要是医学诊断学、制药工业和环境保护。优选地，本发明涉及对抗原和代谢物的同时或先后测定。在本发明的含义中，它们主要是高分子抗原(如蛋白质)或低分子半抗原，例如杀虫剂、新蝶呤、污染物或激素，以及，在代谢物的情况下例如为葡萄糖或肌酸酐。结果可借助于计算图、比色器(参考带)、读数器的帮助从测定法直接确定，或通过用肉眼比较来确定。

Differentiation of human pluripotent stem cells

The present invention provides a method for increasing the expression of MAFA in cells expressing markers characteristic of the pancreatic endocrine lineage comprising the steps of culturing the cells expressing markers characteristic of the pancreatic endocrine lineage in medium comprising a sufficient amount of a cyclin-dependent kinase inhibitor to cause an increase in expression of MAFA.

Pluripotent stem cell culture on micro-carriers

The present invention is directed to methods for the growth, expansion and differentiation of pluripotent stem cells on micro-carriers.

A method for the detection of apoptosis

Methods for the detection of apoptosis by measuring apoptotic bodies shed by apoptotic cells are provided, as are kits to carry out such methods.

A method for measuring the protease activity of c3 and c5 convertase of the alternative complement pathway

Differentiation of human pluripotent stem cells

The present invention provides a method for increasing the expression of MAFA in cells expressing markers characteristic of the pancreatic endocrine lineage comprising the steps of culturing the cells expressing markers characteristic of the pancreatic endocrine lineage in medium comprising a sufficient amount of a cyclin-dependant kinase inhibitor to cause an increase in expression of MAFA.

Sample preparation method for DNA methylation analysis

Improved methods of detecting microbes

Methods of Detecting Viability-Associated Molecules

A method of detecting a molecule associated with viability of one or more cells or organisms in a sample comprises the initial step of contacting the sample with an enzyme, which enzyme is capable of adding or removing a chemical moiety to or from a nucleic acid molecule in the presence of the molecule associated with viability of the of the one or more cells or organisms. This thereby generates a novel detectable nucleic acid molecule. The next step involves detecting the presence of the molecule associated with viability of the one or more cells or organisms by detecting the novel nucleic acid molecule generated only in the presence of the molecule associated with viability of the one or more cells or organisms. A most preferred molecule associated with viability is ATP, although NAD may also be detected. A preferred enzyme for use in the methods is ligase. The method has numerous applications, in particular in monitoring viability of cells, toxicology testing and determining whether there is contamination in a sample or on a surface. Kits are also provided for carrying out the methods.

Kits and Devices for Performing Methods of Detecting Viability-Associated Molecules

Kits and devices for performing a method of detecting a molecule associated with viability of one or more cells or organisms in a sample are provided. The method comprises the initial step of contacting the sample with an enzyme, which enzyme is capable of adding or removing a chemical moiety to or from a nucleic acid molecule in the presence of the molecule associated with viability of the one or more cells or organisms. This thereby generates a novel detectable nucleic acid molecule. The next step involves detecting the presence of the molecule associated with viability of the one or more cells or organisms by detecting the novel nucleic acid molecule generated only in the presence of the molecule associated with viability of the one or more cells or organisms. A most preferred molecule associated with viability is ATP, although NAD may also be detected. A preferred enzyme for use in the methods is ligase. The kits and devices for performing the method have numerous applications, in particular in monitoring viability of cells, toxicology testing and determining whether there is contamination in a sample or on a surface.

Methods of detecting viability-associated molecules

A method of detecting a molecule associated with viability of one or more cells or organisms in a sample comprises the initial step of contacting the sample with an enzyme, which enzyme is capable of adding or removing a chemical moiety to or from a nucleic acid molecule in the presence of the molecule associated with viability of the of the one or more cells or organisms. This thereby generates a novel detectable nucleic acid molecule. The next step involves detecting the presence of the molecule associated with viability of the one or more cells or organisms by detecting the novel nucleic acid molecule generated only in the presence of the molecule associated with viability of the one or more cells or organisms. A most preferred molecule associated with viability is ATP, although NAD may also be detected. A preferred enzyme for use in the methods is ligase. The method has numerous applications, in particular in monitoring viability of cells, toxicology testing and determining whether there is contamination in a sample or on a surface. Kits are also provided for carrying out the methods.

Pluripotent stem cell culture on micro-carriers

본 발명은 마이크로-캐리어 상에서의 만능 줄기 세포의 성장, 증식 및 분화 방법에 관한 것이다. The present invention relates to a method for the growth, proliferation and differentiation of pluripotent stem cells on a micro-carrier.

Method of producing a population of mafa-expressing cells (versions)

FIELD: biotechnology.SUBSTANCE: invention relates to biotechnology, particularly to producing a population of MAFA expressing cells. Method involves differentiation of human cells expressing markers characteristic for line of formed endoderm, obtained from pluripotent stem cells, which are human pluripotent stem cells obtained without destruction of human embryo, human pluripotent stem cells of non-embryonic origin, cells from the human embryonic stem cell line H1, cells from a human embryonic stem cell line H7, cells from a human embryonic stem cell line H9, or cells from a human embryonic stem cell line SA002, into human cells expressing markers characteristic of the pancreatic endoderm line. That is followed by differentiating human cells expressing markers characteristic of a pancreatic endoderm line into human cells expressing markers characteristic of a line of pancreatic endocrine cells expressing at least one of NGN3, NEUROD, ISL1, PDX1, NKX6.1, PAX4 and PTF1-alpha; and treating human cells expressing markers specific to the pancreatic endocrine cell line, with a kinase inhibitor selected from the group consisting of 2-cyanoethyl alster-paullonom; SU9516; alster-pumpallon; Cdk1/2-inhibitor III; casein kinase I inhibitor; Akt-inhibitor IV; EGFR-inhibitor; MEK1/2-inhibitor; staurosporine N-benzoyl-, PKR-inhibitor and PDGF receptor IV tyrosine kinase inhibitor.EFFECT: invention widens the range of agents.21 cl, 16 dwg, 7 tbl, 9 ex РОССИЙСКАЯ ФЕДЕРАЦИЯ (19) RU (11) (13) 2 687 378 C1 (51) МПК C12N 5/071 (2010.01) ФЕДЕРАЛЬНАЯ СЛУЖБА ПО ИНТЕЛЛЕКТУАЛЬНОЙ СОБСТВЕННОСТИ (12) ОПИСАНИЕ ИЗОБРЕТЕНИЯ К ПАТЕНТУ (52) СПК C12N 5/0676 (2019.02); C12N 2501/117 (2019.02) (21) (22) Заявка: 2018111225, 29.03.2018 (24) Дата начала отсчета срока действия патента: 23.10.2009 13.05.2019 31.10.2008 US 61/110,287 Номер и дата приоритета первоначальной заявки, из которой данная заявка выделена: 2014114039 31.10.2008 (45) Опубликовано: 13.05.2019 Бюл. № 14 C 1 2 6 8 7 3 7 8 (54) СПОСОБ ...

DNA ligase assay

A quantitative and functional DNA ligase assay is disclosed. The assay involves the disruption, using restriction enzymes, of a plasmid containing a reporter gene, followed by a ligation step performed in the presence of the biological sample being assayed for DNA ligase activity. The ligation reaction products are then subjected to a coupled transcription-translation reaction, and the extent of DNA ligation is then quantified.

Differentiation of Human Embryonic Stem Cells

The present invention provides a method for increasing the expression of MAFA in cells expressing markers characteristic of the pancreatic endocrine lineage comprising the steps of culturing the cells expressing markers characteristic of the pancreatic endocrine lineage in medium comprising a sufficient amount of a cyclin-dependant kinase inhibitor to cause an increase in expression of MAFA.

Methods for making pancreatic endocrine cells

The present invention provides a method for increasing the expression of MAFA in cells expressing markers characteristic of the pancreatic endocrine lineage comprising the steps of culturing the cells expressing markers characteristic of the pancreatic endocrine lineage in medium comprising a sufficient amount of a cyclin-dependent kinase inhibitor to cause an increase in expression of MAFA.

Ubiquitin ligases, and uses related thereto

The present invention relates to the discovery in eukaryotic cells of ubiquitin ligases. These proteins are referred to herein collectively as "pub" proteins for Protein UBiquitin ligase, and individually as hpub1, h-pub2, h-pub3 and s-publ for the human pub1, pub2 and pub3 and Schizosaccharomyces pombe pub1 clones, repectively. Pub1 proteins apparently play a role in the ubiquitination of the mitotic activating tyrosine phosphatase cdc25, and thus they may regulate the progression of proliferation in eukaryotic cells by activating the cyclin dependent kinase complexes. In S. pombe, disruption of s-pub1 elevates the level of cdc25 protein in vivo increasing the activity of the tyrosine kinases, weel and mik1, required to arrest the cell-cycle. Loss of weel function in an S. pombe cell carrying a disruption in the s-pub1 gene results in a lethal premature entry into mitosis; such lethal phenotype can be rescued by the loss of cdc25 function. A ubiquitin thioester adduct of s-pub1 can be isolated from S. pombe and disruption of s-pub1 dramatically reduces ubiquitination of cdc25.

Method and system for determining a biological response of a target to a soluble candidate substance

A method for determining a biological response of a target (41, 42) to a soluble candidate substance comprises the steps: introducing a soluble candidate substance into a laminar flow of a buffer liquid (2) to form a candidate substance solute (3) having an initial concentration profile (31); dispersing the initial concentration profile (31) to form a dispersed concentration profile (32); directing the dispersed concentration profile (32) into a detection channel (12) to form a final symmetrical concentration profile (33) therein; introducing a target into the detection channel (12) to obtain a combined concentration profile comprising a constant target concentration profile overlying the final symmetrical concentration profile (33); holding in the detection channel (12) at least one half of the combined concentration profile; and optically scanning the combined concentration profile to detect an optical signal representative of the biological response of the target to the soluble candidate substance.

Differentiation of human embrionic stem cells in line of pancreatic endocrine cells

FIELD: medicine, pharmaceutics. SUBSTANCE: invention relates to the field of biotechnology. A method includes stages of cultivating cells, expressing markers, characteristic of a line of pancreatic endocrine cells, in a medium, which contains a sufficient amount of a cyclin-dependent kinase inhibitor to induce increase of MAFA expression. As the cyclin-dependent kinase inhibitor used is ethyl-(6-hydroxy-4-phenylbenzo[4,5]furo[2,3-b])pyridine-carboxylate. The invention can be used in medicine. EFFECT: claimed is the method of increasing MAFA expression in cells, expressing the markers characteristic of the line of the pancreatic endocrine cells. 3 cl, 10 dwg, 7 tbl, 9 ex РОССИЙСКАЯ ФЕДЕРАЦИЯ (19) RU (11) (13) 2 522 001 C2 (51) МПК C12N 5/071 (2010.01) ФЕДЕРАЛЬНАЯ СЛУЖБА ПО ИНТЕЛЛЕКТУАЛЬНОЙ СОБСТВЕННОСТИ (12) ОПИСАНИЕ (21)(22) Заявка: ИЗОБРЕТЕНИЯ К ПАТЕНТУ 2011121842/10, 23.10.2009 (24) Дата начала отсчета срока действия патента: 23.10.2009 (72) Автор(ы): РЕЗАНИА Алиреза (US), ФРАЙЕР Бенджамин (US) (73) Патентообладатель(и): СЕНТОКОР ОРТО БАЙОТЕК ИНК. (US) Приоритет(ы): (30) Конвенционный приоритет: (43) Дата публикации заявки: 10.12.2012 Бюл. № 34 R U 31.10.2008 US 61/110,287 (45) Опубликовано: 10.07.2014 Бюл. № 19 (85) Дата начала рассмотрения заявки PCT на национальной фазе: 31.05.2011 C 2 C 2 dependent kinase 5 activity protects pancreatic beta cells from glucotoxicity", J Biol Chem. 2006 Sep 29;281(39):28858-64. Epub 2006 Aug 3. WO 2006138433 A2 (UNIV CALIFORNIA), 28.12.2006. RU 2333243 C2 (БЛАСТИКОН БИОТЕХНОЛОГИШЕФОРШУНГГМБХ), 10.09.2009 (86) Заявка PCT: 2 5 2 2 0 0 1 US 2009/061774 (23.10.2009) R U 2 5 2 2 0 0 1 (56) Список документов, цитированных в отчете о поиске: UBEDA M et al., "Inhibition of cyclin- (87) Публикация заявки PCT: WO 2010/051223 (06.05.2010) Адрес для переписки: 129090, Москва, ул. Б.Спасская, 25, стр.3, ООО "Юридическая фирма Городисский и Партнеры", пат.пов. Е.Е.Назиной (54) ДИФФЕРЕНЦИРОВАНИЕ ЧЕЛОВЕЧЕСКИХ ЭМБРИОНАЛЬНЫХ СТВОЛОВЫХ КЛЕТОК В ...

Novel diagnostic marker for type I diabetes mellitus

본 발명은 제1형 당뇨병의 신규한 진단 마커에 관한 것으로, 알라닌 tRNA 합성효소(alanyl-tRNA synthetase), 글리신 tRNA 합성효소(glycyl-tRNA synthetase), 아스파라긴 tRNA 합성효소(asparaginyl-tRNA synthetase) 또는 트립토판 tRNA 합성효소(tryptophanyl-tRNA synthethase)를 포함하는 제1형 당뇨병 진단용 조성물, 이를 포함하는 진단 키트 및 진단 방법을 제공한다. 본 발명의 조성물, 키트 및 방법은 환자의 시료로부터 손쉽게 제1형 당뇨병 여부를 진단할 수 있으므로 제1형 당뇨병의 조기 진단 및 확정을 위하여 사용할 수 있다. The present invention relates to novel diagnostic markers of type 1 diabetes, including alanine-tRNA synthetase, glycine-tRNA synthetase, asparaginyl-tRNA synthetase or tryptophan Provided is a composition for diagnosing type 1 diabetes comprising a tRNA synthetase (tryptophanyl-tRNA synthethase), a diagnostic kit including the same, and a diagnostic method. The compositions, kits, and methods of the present invention can be easily diagnosed from type 1 diabetes from a patient's sample and can be used for early diagnosis and confirmation of type 1 diabetes. 아미노아실 tRNA 합성효소, 제1형 당뇨병, 진단, 마커 Aminoacyl tRNA synthetase, type 1 diabetes, diagnostic marker

Sample preparation method for dna methylation analysis

The invention provides a method for DNA methylation analysis using bisulfite treatment, wherein the method is characterized in that the yield of DNA sample that can be amplified by PCR after bisulfite treatment is increased by treating the DNA sample after bisulfite treatment with a single-stranded DNA ligase.

Mass spectrometry assays for identifying compounds that activate deacetylases

Provided are methods for determining the activity of proteins that modulate the acetylation state of a protein substrate. The methods may be used for determining both acetyltransferase activity and deacetylase activity. The methods utilize mass spectrometry for determining the acetylation state of a substrate peptide. The methods may also be used to identify compounds that modulate the activity of a protein having acetyltransferase or deacetylase activity. In some embodiments, a compound that modulates a deacetylase is an activator of the deacetylase.

Non-oligomerizing tandem fluorescent proteins

Non-oligomerizing fluorescent proteins, which are formed by operatively linking two or more monomers of a fluorescent protein, or which are derived from a fluorescent protein having at least one mutation that reduces or eliminates the ability of the fluorescent protein to oligomerize, are provided. The non-oligomerizing fluorescent proteins can be derived from a naturally occurring green fluorescent protein, a red fluorescent protein, or other fluorescent protein, or a fluorescent protein related thereto. Also provided is a fusion protein, which includes a non-oligomerizing fluorescent protein linked to at least one polypeptide of interest. In addition, a polynucleotide encoding a non-oligomerizing fluorescent protein is provided, as is a recombinant nucleic acid molecule, which includes polynucleotide encoding a non-oligomerizing fluorescent protein operatively linked to at least a second polynucleotide. Vectors and host cells containing such polynucleotides also are provided, as are kits containing one or more non-oligomerizing fluorescent proteins or encoding polynucleotides or constructs derived therefrom. Further provided are methods of making and using the proteins and polynucleotides.

Method of detecting thrombosis by measuring von willebrand factor-cleaving protease

A method of detecting the level of thrombus formation tendency or thrombosis through determining of the quantity of von Willebrand factor splitting enzyme. There is further disclosed a kit for detecting the level of thrombus formation tendency or thrombosis, which kit includes an antibody capable of specific binding to von Willebrand factor splitting enzyme, or a fragment of the antibody. The provided detection method and detection kit excel in convenience, speed and specificity.

Novel diagnostic marker for type 1 diabetes mellitus

本发明涉及第一型糖尿病的新型诊断标志物，氨酰基tRNA合成酶或是包含对于这抗体的有效成分的第一型糖尿病的用于诊断的组成物，氨酰基tRNA合成酶的第一型糖尿病诊断用制剂的制造或从个体的样品中抗-丙氨酸抗体合成的元素，抗-甘氨酸tRNA合成酶抗体，抗-天冬酰胺tRNA抗体合成的元素或抗-色氨酸tRNA检测到的抗体合成的元素的第1型糖尿病的诊断的方法。本发明是试剂工具组成或方法是从对患者的治疗更容易诊断出第1型糖尿病的早期或确定的用处。

Application of ubiquitin ligase CHAF1B as target site in preparation of lung adenocarcinoma cisplatin sensitization drug

本发明公开了泛素连接酶CHAF1B作为靶位点在制备肺腺癌顺铂增敏药物中的应用。本发明研究表明，肺腺癌顺铂耐药细胞株中泛素连接酶CAHF1B高表达，予以敲除CAHF1B后可增加肺腺癌细胞的顺铂药物敏感性。因此，CAHF1B可在制备肺腺癌顺铂增敏药物中作为靶位点，也是临床上预测肺腺癌患者顺铂药物敏感性的的分子标志物。机制研究表明，高表达的CAHF1B可通过促进NCOR2泛素化降解导致肺腺癌细胞顺铂耐药。因此，NCOR2也在制备肺腺癌顺铂增敏药中作为靶位点。

Methods for detecting or quantifying ctp and ctp synthase activity

The present invention relates to a method for detecting or quantifying CTP in a cell sample comprising at least two nucleotide triphosphates by cationic ion pairing chromatography coupled to mass spectrometry, to a method for detecting or quantifying CTP synthase activity based on the method for detecting or quantifying CTP, and to their use in methods for screening potential immunosuppressive or anti-cancer compounds and in methods for determining the appropriate dose of an immunosuppressive or anti-cancer compound inhibiting CTP synthase activity for a treated subject.

Ubiquitin E3 ligase

The present invention provides a native or reconstituted complex comprising Bmi1 and/or Ring1 and/or Ring2, wherein the complex has ubiquitin E3 ligase activity. Optionally, the complex further comprises HPH2 and/or HPC3. Also disclosed are methods of producing the reconstituted complex, methods of identifying compounds that modulate the ubiquitin E3 ligase activity of the native or reconstituted complex, and methods of identifying candidate compounds for treating cancer. Kits for determining modulation of protein ubiquitination and/or for ubiquitinating a target substrate are further provided.

Non-oligomerizing tandem fluorescent proteins

Method for judging whether anaerobic digestion process enters stable stage

本发明提供了一种厌氧消化过程进入稳定阶段的判定方法，包括：启动厌氧消化过程至少3天后，每间隔预设时间从所述反应体系中取一定量液态反应物并测量所取液态反应物中的甲基辅酶M的含量；当至少连续15天内所取液态反应物中的甲基辅酶M的含量持续增加时，确定厌氧过程消化进入稳定阶段。该方法测量对象为液体，比测量气体稳定性更高，且通过试剂盒即可进行测量，对人员经验要求低、安全可靠。

Improvements in or relating to growth of microorganisms

Pluripotent stem cell culture on micro-carriers

본 발명은 마이크로-캐리어 상에서의 만능 줄기 세포의 성장, 증식 및 분화 방법에 관한 것이다.

Protein phosphorylation in t-cell receptor signaling pathways

The invention discloses 95 novel phosphorylation sites identified in signal transduction proteins and pathways downstream of the T-cell receptor, and provides phosphorylation-site specific antibodies and heavy-isotope labeled peptides (AQUA peptides) for the selective detection and quantification of these phosphorylated sites/proteins, as well as methods of using the reagents for such purpose. Among the phosphorylation sites identified are sites occurring in the following protein types: Actin Binding proteins, Adaptor/Scaffold proteins, Adhesion proteins, Calcium-binding proteins, Cell Cycle Regulation or Channel proteins, Chaperones, Cofactor proteins, Cytoskeletal proteins, DNA Binding proteins, G protein or GTPase Activating proteins, Ligases, Lipid Kinases and Binding proteins, Oxidoreductases, Protein Kinases, Protein Phosphatases, Receptor proteins, RNA Binding proteins, Transcription Factor/Initiation Complex proteins, Transcription Coactivator/Corepressor proteins, Translation Initiation Complex proteins, Ubitquitin Conjugating System proteins, and Vesicle proteins.

Differentiation of human embryonic stem cells to the pancreatic endocrine lineage.

La presente invención provee un método para aumentar la expresión de MAFA en las células que expresan los marcadores característicos del linaje endócrino pancreático que comprende los pasos de cultivar las células que expresan los marcadores característicos del linaje endócrino pancreático en un medio que comprende una cantidad suficiente de un inhibidor de quinasa que depende de la cíclina para causar un aumento en la expresión de MAFA.

Non-oligomerizing tandem fluorescent proteins

Non-oligomerizing fluorescent proteins, which are formed by operatively linking two or more monomers of a fluorescent protein, or which are derived from a fluorescent protein having at least one mutation that reduces or eliminates the ability of the fluorescent protein to oligomerize, are provided. The non-oligomerizing fluorescent proteins can be derived from a naturally occuring green fluorescent protein, a red fluorescent protein, or other fluorescent protein, or a fluorescent protein related thereto. Also provided is a fusion protein, which includes a non-oligomerizing fluorescent protein linked to at least one polypeptide of interest. In addition, a polynucleotide encoding a non-oligomerizing fluorescent protein is provided, as is a recombinant nucleic acid molecule, which includes polynucleotide encoding a non-oligomerizing fluorescent protein operatively linked to at least a second polynucleotide. Vectors and host cells containing such polynucleotides also are provided, as are kits containing one or more non- oligomerizing fluorescent proteins or encoding polynucleotides or constructs derived therefrom. Further provided are methods of making and using the proteins and polynucleotides

Method for detection of the presence or absence of methylthioadenosine phosphorylase (MTAse) in a cell sample by detection of the presence or absence of MTAse encoding nucleic acid in the cell sample

A method for detecting whether methyladenosine phosphatase (MTAse) is present in a cell sample. In one respect, the method comprises adding oligonucleotide probes to the sample, which probes are capable of specifically hybridizing to any MTAse encoding nucleic acid in the sample under conditions favoring that hybridization. Absence of MTAse in a sample is considered to be indicative of malignancy. Polynucleotides encoding MTAse, MTAse peptides and antibodies to MTAse, as well as kits for performing the methods of the invention, are provided.

Method of detecting thrombosis by measuring von willebrand factor-cleaving protease

A method of detecting the level of thrombus formation tendency or thrombosis through determining of the quantity of von Willebrand factor splitting enzyme. There is further disclosed a kit for detecting the level of thrombus formation tendency or thrombosis, which kit includes an antibody capable of specific binding to von Willebrand factor splitting enzyme, or a fragment of the antibody. The provided detection method and detection kit excel in convenience, speed and specificity.

Fungi resistant to soraphen a

P27 ubiquitination assay and methods of use

This invention relates to easy and reliable assays for the ubiquitination of p27. Because the assay can be performed as a plate capture assay, or as a homogenous time-resolved fluorescence resonance energy transfer assay, with defined, easily replicatable reaction conditions, it is particularly useful for high-throughput screening of, for example, a prospective anticancer agent. The invention provides methods to determine the amount of ubiquitination of p27, or of any protein, and use of the method to identify compounds that modulate the ubiquitination of p27, or any protein.

Method of identifying polyubiquitinated substrate

Monomeric and dimeric fluorescent protein variants and methods for making same

The present invention relates generally to variant fluorescent proteins, and more specifically to monomeric and dimeric forms of Anthozoan fluorescent proteins. In one aspect, the present invention provides variants of fluorescent proteins, where the variants have a reduced propensity to tetramerize, and form dimeric or monomeric structures. The invention also relates to methods of making and using such fluorescent protein monomers and dimers.

Novel diagnostic marker for type 1 diabetes mellitus

The present invention provides, as a novel diagnosis marker for type 1 diabetes mellitus, a type 1 diabetes mellitus diagnostic composition comprising alanyl-tRNA synthetase, glycyl-tRNA synthetase, asparaginyl-tRNA synthetase, or tryptophanyl-tRNA synthetase, a diagnostic kit comprising the same, and a diagnostic method using the same. The composition, the kit, and the method, according to the present invention, may be used for early diagnosis and confirmed diagnosis of type 1 diabetes mellitus because type 1 diabetes mellitus can be easily diagnosed from a patient sample.

E3 ubiquitin ligase (ube3a) protein targets

The invention relates to UBE3A protein targets and their usage as target engagement biomarkers for compounds that modulate ube3a expression.

Methods for rapidly transforming monocots

The present invention provides methods for transforming monocot plants via a simple and rapid protocol, to obtain regenerated plants capable of being planted to soil in as little as 4-8 weeks. Associated cell culture media and growth conditions are also provided, as well as plants and plant parts obtained by the method. Further, a method for screening recalcitrant plant genotypes for transformability by the methods of the present invention is also provided. Further, a system for expanding priority development window for producing transgenic plants by the methods of the present invention is also provided.