Нитроны стероидов для лечения и предотвращения инсульта или ишемии головного мога, болезни Альцгеймера, болезни Паркинсона и бокового амиотрофического склероза

Группа изобретений относится к фармакологии и медицине. Предложены: применение нитрона стероида, выбранного из соединений (E)-N-((8S,9S,10R,13R,14S,17R)-10,13-диметил-17-((R)-6-метилгептан-2-ил)-7,8,9,11,12,13,14,15,16,17-декагидро-1Н-циклопента[а]фенантрен-3(2Н,6Н,10Н)-илиден)метанамин оксида (F2) и (Z)-N-((8S,9S,10R,13R,14S,17R)-10,13-диметил-17-((R)-6-метилгептан-2-ил)-7,8,9,11,12,13,14,15,16,17-декагидро-1Н-циклопента[а]фенантрен-3(2Н,6Н,10Н)-илиден)метанамин оксида (F3), или фармацевтически приемлемых солей и гидратов указанных соединений для производства фармацевтической композиции или лекарственного препарата для предотвращения и/или лечения инсульта или ишемии головного мозга, болезни Альцгеймера, болезни Паркинсона и бокового амиотрофического склероза; композиция на их основе нейропротекторного действия, применение указанной композиции в сочетании с тромболитическим агентом для лечения инсульта или ишемии головного мозга, болезни Альцгеймера, болезни Паркинсона и бокового амиотрофического ...

ОЛИГОНУКЛЕОТИДЫ ДЛЯ ИНДУЦИРОВАНИЯ ОТЦОВСКОЙ ЭКСПРЕССИИ UBE3A

Изобретение относится к области биотехнологии. Описана группа изобретений, включающая антисмысловой олигонуклеотид, индуцирующий экспрессию UBE3A в клетке-мишени, где подавляется экспрессия отцовского UBE3A, фармацевтическую композицию, содержащую вышеуказанный антисмысловой олигонуклеотид, способ индуцирования экспрессии UBE3A в клетке-мишени, где подавляется экспрессия отцовского UBE3A, применение антисмыслового олигонуклеотида или фармацевтической композиции в лечении или предупреждении синдрома Ангельмана и применение антисмыслового олигонуклеотида или фармацевтической композиции для приготовления лекарственного средства для применения в лечении или предупреждении синдрома Ангельмана. Антисмысловой олигонуклеотид согласно настоящему изобретению представляет собой олигоунклеотидное соединение TTAcActtaattatactTCC, где прописные буквы представляют нуклеозиды бета-D-окси LNA, строчные буквы представляют нуклеозиды ДНК, все С LNA представляют собой 5-метилцитозин и все межнуклеозидные связи ...

ВИРИОНЫ АДЕНОАССОЦИИРОВАННОГО ВИРУСА С ВАРИАНТНЫМ КАПСИДОМ И СПОСОБЫ ИХ ИСПОЛЬЗОВАНИЯ

Изобретение относится к биотехнологии. Описан вирион рекомбинантного аденоассоциированного вируса rAAV, содержащий: a) капсидный белок вариантного AAV, который содержит пептидную вставку, по сравнению с соответствующим капсидным белком родительского AAV, где пептидная вставка имеет длину от 7 аминокислот до 10 аминокислот, где участок вставки расположен между двумя соседствующими аминокислотами в положении между аминокислотами, соответствующем аминокислотам 570 и 611 VP1 из AAV2 или соответствующем положении в капсидном белке другого серотипа AAV; и b) гетерологичную нуклеиновую кислоту, содержащую нуклеотидную последовательность, кодирующую генный продукт. Представлена фармацевтическая композиция, содержащая данный вирион. Представлен способ лечения заболевания сетчатки, который включает введение индивидууму, нуждающемуся в этом, эффективного количества описанного вириона. При осуществлении изобретения достигается большая инфекционность ретинальной клетки, поскольку вирионы AAV вводятся ...

СПОСОБ ПОЛУЧЕНИЯ ИНДУЦИРОВАННЫХ НЕРВНЫХ СТВОЛОВЫХ КЛЕТОК, ПЕРЕПРОГРАММИРОВАННЫХ ИЗ КЛЕТОК, НЕ ЯВЛЯЮЩИХСЯ НЕРВНЫМИ, С ИСПОЛЬЗОВАНИЕМ HMGA2

СРЕДСТВА И СПОСОБЫ ДЛЯ ОПРЕДЕЛЕНИЯ БИОЛОГИЧЕСКОЙ АКТИВНОСТИ ПОЛИПЕПТИДОВ НЕЙРОТОКСИНА В КЛЕТКАХ

СПОСОБ И УСТРОЙСТВО ДЛЯ ИЗГОТОВЛЕНИЯ ФОРМОВАННЫХ ПОРЦИЙ МЯСА ИЗ ЦЕЛЬНЫХ КУСКОВ НАТУРАЛЬНОГО МЯСА

... 1. Способ изготовления формованных порций мяса из цельных кусков натурального мяса, в частности из мяса крупного рогатого скота, свиней, овец, коз, дичи, птицы или рыб всех видов, посредством следующих технологических операций: подача цельных кусков натурального мяса на технологическую линию переработки; отбор тех кусков мяса, вес которых меньше, чем заданный вес порций мяса; обрезка тех кусков мяса, вес которых превышает заданный вес порций мяса; отбор тех кусков мяса, вес которых соответствует заданному весу; замораживание отобранных кусков мяса с получением формованных порций мяса; формирование формованных порций мяса из замороженных порций мяса путем закладки в формовочный лоток и спрессовывания или же путем спрессовывания и формования штамповкой в формовочном лотке; причем отобранные куски мяса или наборы из мелких кусков мяса помещают перед замораживанием в форму, размеры основания которой равны или же меньше, чем размеры основания формовочного лотка; и регулируемым образом, посредством ...

СПОСОБЫ ИДЕНТИФИКАЦИИ, ИЗОЛЯЦИИ И ПРИМЕНЕНИЯ РЕЦЕПТОРОВ ПАХУЧИХ ВЕЩЕСТВ И АРОМАТОВ

МИКРООРГАНИЗМЫ И ИХ ФРАКЦИИ, ИНДУЦИРУЮЩИЕ СПЕЦИФИЧНЫЙ К УГЛЕВОДУ КЛЕТОЧНЫЙ ИММУНИТЕТ

... 1. Способ выделения положительного по углеводу микроорганизма, содержащего интересующий углеводный эпитоп, из смеси микроорганизмов, включающий ! (a) создание контакта между связывающей углевод молекулой, специфичной к интересующему углеводному эпитопу, и смесью микроорганизмов, и ! (b) выделение как минимум одного микроорганизма, связавшегося с указанной связывающей углевод молекулой, из указанной смеси, ! (c) необязательную проверку того, что выделенный микроорганизм является положительным по углеводу микроорганизмом, путем тестирования выделенного микроорганизма на специфическое связывание с указанной связывающей углевод молекулой. ! 2. Способ по п.1, дополнительно включающий (d) тестирование индукции эффективного специфичного к углеводу клеточного иммунного ответа и/или гуморального иммунного ответа против указанного углеводного эпитопа указанным микроорганизмом и/или его фракцией и/или лизатом in vivo или in vitro. ! 3. Способ по п.2, где этап (d) включает тестирование индукции эффективного ...

АНТИТЕЛО ПРОТИВ ПРОТЕИНТИРОЗИНФОСФАТАЗЫ σ РЕЦЕПТОРНОГО ТИПА ЧЕЛОВЕКА

СИСТЕМЫ

Нитроны стероидов для лечения и предотвращения инсульта или ишемии головного мога, болезни Альцгеймера, болезни Паркинсона и бокового амиотрофического склероза

VERFAHREN UND ZUSAMMENSETZUNGEN ZUR BEOBACHTUNG DER ZELLULÄREN VERARBEITUNG VON BETA-AMYLOID-VORLÄUFERPROTEINEN

Testsystem für die Findung von Wirkstoffen gegen prioneninduzierte Erkrankungen und Wirkstoffe zur Verhütung und zur Behandlung dieser Erkrankung

Die Erfindung betrifft ein Testsystem zur Auffindung von Wirksubstanzen, welche die Hemmung des Axonwachstums durch infektiöse Prionen aufheben, mit den folgenden Komponenten: a) einer in Kultur gehaltenen Nervenzelle, wobei diese Nervenzelle in der Zellkultur Axone ausbildet, b) eine Zubereitung, enthaltend abnormales Prion-Protein, und c) ein Nachweissystem für die Ausbildung von Axone, ein Verfahren zur Auffindung von Wirksubstanzen, welche die Hemmung des Axonwachstums durch infektiöse Prionen aufheben, mittels eines solchen Testsystems, sowie damit ermittelbare Wirksubstanzen.

METHODE UND REAGENZIEN ZUM IDENTIFIZIEREN VON MODULATOREN NEURONALER APOPTOSE

Method of inducing memory B cell development and terminal differentiation

Neurodegenerative disorders

Human N-methyl-D-aspartate receptor subunits,nucleic acids encoding same and uses therefor

Device for forming a cellular network

CNS CHLORIDE MODULATING AND USE THE SAME

USE OF A BIOLOGICAL SENSOR TO THE PROOF OF ANTIBIOTICS

HUMAN METABOTROPI RECEPTORS, THIS CODING NUCLEIC ACID AND YOUR USE

NOGO RECEPTOR-OBTAINED BLOCKADE THE AXONALEN OF GROWTH

MARKER FOR THERAPY WITH ANTIDEPRESSIVES AND THUS IN CONNECTION STANDING PROCEDURE

COMPOSITIONS FOR THE STIMULATION OF THE REGENERATION AND THE RE-ESTABLISHMENT OF THE NERVOUS SYSTEM BY ADJUSTMENT THE ARGINASE 1 - AND POLYAMINE SYNTHESIS

RECOGNITION OF TRANSMEMBRANPOTENTIALEN BY OPTICAL PROCEDURES

SYSTEM FOR THE MEASUREMENT OF THE ELECTRICAL POTENTIAL BY A DIAPHRAGM

MOLECULES, ITSELF IN SELECTED ORGANS OR FABRICS THE INVIVO APPEARING AND PROCEEDING FOR YOUR IDENTIFICATION

SCREENINGVERFAHREN FROM MORPHOGENI PROTEINS

Marker for neural stem cells

The present application concerns methods for detecting and isolating a population of neural stem cells (NSC) or neural progenitor cells (NPC) based on expression of the marker integrin alpha10beta1; as well as use of said population of NSC or NPC for therapy, diagnosis and prognosis of disease and damage of the CNS.

Oligonucleotides for inducing paternal UBE3A expression

The present invention relates to oligonucleotides that are capable of inducing expression of ubiquitin-protein ligase E3A (UBE3A) from the paternal allele in animal or human neurons. The oligonucleotides target the suppressor of the UBE3A paternal allele by hybridization to SNHG14 long non-coding RNA downstream of SNORD109B. The present invention further relates to pharmaceutical compositions and methods for treatment of Angelman syndrome.

NEURODEGENERATIVE DISORDERS

... - 55 Abstract A cyclic polypeptide, derivative or analogue thereof, comprising an amino acid sequence derived from the C-terminus of acetylcholinesterase (AChE), or a truncation thereof. [Figure: 11] Figure 11 A DAEFR HDSGY EVHHQ KLVFF AEDVG SNKGA IIGLM VGGVV IA T30 KAEFH RWSSY MVHWK NQFDK YSKQD RCSDL T14 AEFH RWSSY MVHWK TI NQFDH YSKQD RCSDL Figure 12 Orithosteric site Alosteric site MLAACh, Chol 0 PNU12 IVM, GAL, T30 Figure 13 Calcium influx .'z 100- - - - - - - - - -3 1 0 Log [NBP-14] ...

Inhibition of tau-tau-association

METHODS OF TREATING NEUROLOGICAL CONDITIONS WITH HEMATOPOEITIC GROWTH FACTORS

Identification of pain-regulated gene in the dorsal root ganglion (drg) after cfa-induced arthritis

Benzimidazole-carboxamide compounds as 5-HT4 receptor agonists

Microtubule synthesis as a biomarker

Methods of treating neurological conditions with hematopoeitic growth factors

Methods of suppressing microglial activation

Method for Screening Transglutaminase 2 Inhibitor or Activator

Compositions comprising ADDL receptor syngap

Methods and materials relating to enhanced production of dopamine neurons

Multi-parameter high throughput screening assays (mphts)

Use of non-immunosuppressive compounds for promoting nerve regeneration

Methods and compositions to treat pain and painful disorders using 577, 20739 or 57145

A non-selective cation channel in neural cells and methods for treating brain swelling

A CELLULAR ASSAY SYSTEM FOR NEUROFIBRILLARY TANGLE FORMATION

Metabolism of SOD1 in CSF

The disclosure relates to methods for the diagnosis and treatment of neurological and neurodegenerative diseases, disorders, and associated processes. Specifically, the disclosure relates to a method for measuring the metabolism of central nervous system derived biomolecules in a subject in vivo. Further disclosed are methods for measuring the in vivo metabolism (e.g. the rate of synthesis, the rate of clearance) of neurally derived biomolecules, such as superoxide dismutase 1 (SOD1 ).

Anti-human receptor-type protein tyrosine phosphatase sigma antibody

H:\fm\Intrwovn\NRPortbl\DCC\FMT\I2080439 L.docx-11/16/2016 A monoclonal antibody that binds to an extracellular domain of human receptor-type protein tyrosine phosphatase Y (human PTPRS), or a fragment including an antigen-binding region thereof.

Peripheral zone tumor cells, methods for their preparation and use

Compositions and methods for modulating neuronal excitability and motor behavior

The invention is directed to compositions and methods for treating or reducing the likelihood of the development of epilepsy in an individual. The method comprises administering to the central nervous system of an individual in need of such treatment a therapeutically effective amount of an agent capable of increasing the expression and/or activity of miR-128.

Methods for the identification, assessment, prevention, and treatment of neurological disorders and diseases using Fndc5

The invention provides methods for identifying, assessing, preventing, and treating neurological disorders and diseases using Fndc5 and modulators of Fndc5 expression or activity.

A non-selective cation channel in neural cells and methods for treating brain

A composition comprising a novel Ca2-activated, [ATP]-sensitive nonspecific cation (NCca ATP) channel is described. The channel is found in mammalian neural cells and exhibits a different sensitivity to block by various adenine nucleotides, and is activated by submicromolar [Ca]i. The NCCa-ATP channel is activated under conditions of ATP depletion, which causes severe cell depolarization, followed by cell swelling- The NCCa-ATP channel is regulated by a sulfonylurea receptor and is inhibited by sulfonylurea compounds glibenclamide and tolbutamide. Methods employing compositions comprising the NCCa-ATP channel to screen for compounds that block the channel and the use of such antagonists as therapeutics in preventing brain swelling and damage are described. In addition, methods employing compositions comprising the Kir2.3 channel to screen for compounds that open the channel and the use of such antagonists as therapeutics in preventing brain swelling and damage are described.

Production method for retinal tissue

The present invention provides a production method for retinal cells or retinal tissue, the method including steps (1)-(3): (1) a first step for culturing human pluripotent stem cells in the absence of feeder cells in a culture medium that includes a non-differentiation maintenance factor; (2) a second step for culturing the pluripotent stem cells obtained in the first step in suspension in the presence of a sonic hedgehog signal transduction pathway activator so as to form an aggregate of cells; and (3) a third step for culturing the aggregate obtained in the second step in suspension in the presence of a BMP signal transduction pathway activator so as to obtain an aggregate that includes retinal cells or retinal tissue.

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

GENERATION OF NEURAL STEM CELLS FROM HUMAN TROPHOBLAST STEM CELLS

... [00384] Provided herein are isolated neural stem cells. Also provided are methods for treatment of neurodegenerative diseases using suitable preparations comprising the isolated neural stem cells.

Method to diagnose and monitor cellular immune deficiencies

Pgt and apoptosis

METHODS AND REAGENTS FOR IDENTIFYING SYNTHETIC GENETIC ELEMENTS

The present invention relates to a selection method that allows fast recovery and identification of functional gene fragments which selectively inhibit growth, e.g., are cytostatic or cytotoxic, of particular cell-types, such as transformed cells. The strategy relies, in part, on the ability of small gene fragments to encode dominant-acting synthetic genetic elements (SGEs), e.g., molecules which interfere with the function of genes from which they are derived. SGEs which can be identified by the subject method include, but are not limited to, inhibitory antisense RNA molecules, ribozymes, nucleic acid decoys, and small peptides.

LGI, LINGO AND P75NTR FAMILY MEMBERS: NOVEL MODULATORS OF NEURONAL GROWTH

The present invention relates to a novel method to promote regeneration or repair of the central or peripheral nervous system following injury The present invention concerns the use of a leucine-rich, glioma- inactivated protein (LGIn), or an analog or derivative thereof, to promote the regeneration or remyelination of neurons after injury to the central nervous system LGIns are endogenous proteins secreted by central neurons that promote regeneration of neurons after injury to the central nervous system The present invention includes an assay to measure the interaction of LGIn with LINGOn and p75NTRn as well as to identify factors that enhance or disrupt these interactions The invention further includes cell lines capable of expressing LGIn, LINGOn and p75NTRn molecules, as well as the proteins purified from these cells.

METHODS FOR SCREENING INHIBITORS OF APOPTOSIS

The present invention addresses a need in the art for methods of identifying apoptotic proteins and methods for screnning compounds which inhibit an poptotic protein. More particularly, in certain embodiments, the invention relates to increased expression levels of a NALP1 gene and/or a NALP5 gene following neuron injury. In other embodiments, the present invention demonstrates that the recombinant expression of NALP1 and/or NALP polynucleotides stimulates apoptosis in cultured neurons, Hela cells and NIH- 3T3 cells. In yet other embodiments, the invention relates to mutations in the nucleotide binding sequence (NBS) of a NALP or a NALP5 polypeptide, wherein these NBS mutations inhibit purine nucleotide binding and reduce caspase activation.

METHODS OF SUPPRESSING MICROGLIAL ACTIVATION AND SYSTEMIC INFLAMMATORY RESPONSES

Methods of suppressing the activation of microglial cells in the Central Nervous System (CNS), methods of ameliorating or treating the neurological effects of cerebral ischemia or cerebral inflammation, and methods of combating specific diseases that affect the CNS by administering a compound that binds to microglial receptors and prevents or reduces microglial activation are described. ApoE receptor binding peptides that may be used in the methods of the invention are also described, as are methods of using such peptides to treat peripheral inflammatory conditions such as sepsis. Also described are methods of screening compounds for the ability to suppress or reduce microglial activation.

COMPOUND ASSAY USING NEMATODES

The invention provides methods of screening compounds for potential pharmacological activity using nematode worms, principally but not exclusively the nematode C. elegans. Specifically, the invention relates to the use of nematodes modified to have certain characteristics which provide advantages for compound screening, such as constitutive pharyngeal pumping, increased gut permeability or altered gut molecular transport. Methods for selecting suitably modified nematodes from a population of nematodes are also provided.

MODULATORS OF ODORANT RECEPTORS

... ²²²The present invention relates to polypeptides capable of promoting odorant ²receptor cell surface localization and odorant receptor functional expression. ²The present invention further provides assays for the detection of ligands ²specific for various odorant receptors. Additionally, the present invention ²provides methods of screening for odorant receptor accessory protein ²polymorphisms and mutations associated with disease states, as well as methods ²of screening for therapeutic agents, ligands, and modulators of such proteins.² ...

GABA NEURON PROGENITOR CELL MARKER 65B13

In the spinal dorsal horn and the cerebellum, a selective marker gene 65B 13 for a GABA neuron precursor cell can be identified, and a GABA neuron pre cursor cell can be isolated by using an antibody capable of binding to a pro tein encoded by the gene. It is found that 65B13 is useful as a marker for i solating a GABA-producing neuron precursor cell in the spinal dorsal horn or the cerebellum. By employing the identified marker as an indicator, a GABA neuron precursor cell can be identified or isolated efficiently.

METHODS OF SUPPRESSING MICROGLIAL ACTIVATION AND SYSTEMIC INFLAMMATORY RESPONSES

... ²²²Methods of suppressing the activation of microglial cells in the Central ²Nervous System (CNS), methods of ameliorating or treating the neurological ²effects of cerebral ischemia or cerebral inflammation, and methods of ²combating specific diseases that affect the CNS by administering a compound ²that binds to microglial receptors and prevents or reduces microglial ²activation are described. ApoE receptor binding peptides that may be used in ²the methods of the invention are also described, as are methods of using such ²peptides to treat peripheral inflammatory conditions such as sepsis. Also ²described are methods of screening compounds for the ability to suppress or ²reduce microglial activation.² ...

COMPOSITIONS AND METHODS FOR TREATING NEUROLOGICAL DISORDERS OR DAMAGE

The invention relates to a clonogenic neurosphere assay to carry out high throughput screens (HTS) to identify potent and/or selective modulators of proliferation, differentiation and/or renewal of neural precursor cells, neural progenitor cells and/or self-renewing and multipotent neural stem cells (NSCs). The invention also relates to compositions comprising the identified modulators and methods of using the modulators and compositions, in particular to treat neurological disorders (e.g. brain or CNS cancer) or damage.

GREEN TEA POLYPHENOL ALPHA SECRETASE ENHANCERS AND METHODS OF USE

The subject invention concerns materials and methods for treating or preventing a neurodegenerative condition or disease associated with .beta.- amyloid peptide deposition in neural tissue in a person or animal by administering a therapeutically effective amount of a polyphenol, or an analog, isomer, metabolite, or prodrug thereof, that increases expression or activity of a protein that exhibits .alpha.-secretase activity. The subject invention also provides methods to increase .alpha.-secretase expression and/or activity in cells by administering polyphenol flavonoids like (-)- epigallocatechin-3~gallate (EGCG) and epicatechin (EC), two polyphenols derived from green tea and other plants and that can be produced synthetically. Furthermore, there are provided methods to decrease or inhibit the production of A.beta.1-40 or A.beta.1-42 by administering the EGCG and EC compounds, their analogs, metabolites, and prodrugs.

COMPOUNDS AND COMPOSITIONS FOR TREATING TISSUE ISCHEMIA

There is disclosed a genus of compounds and pharmaceutical compositions that are protective for mitigating damage associated with tissue ischemia, particularly stroke (CNS ischemia), and ischemia of the myocardium. The present invention further provides a method for treating tissue damage caused by ischemia. Lastly, the present invention provides a method for treating tissue damage caused by providing a compound that inhibits the cytotoxic activity of 3-aminopropanal.

PERSONALIZED 3D NEURAL CULTURE SYSTEM FOR GENERATING HUMAN OLIGODENDROCYTES AND STUDYING MYELINATION IN VITRO

Human pluripotent stem cells are differentiated in vitro into oligodendro-spheroids comprising oligodendrocytes for use in analysis, screening programs, and the like.

COMPOUNDS FOR TREATMENT OF DISEASES AND METHODS OF SCREENING THEREFOR

The present invention related to a compound for use in prevention or treatment of a neurodegenerative disease associated with the formation of stress granules. The compound is selected from lipoic acid, lipoamide, dihydrolipoic acid, and dihydrolipoamide. The invention further relates to a method of identifying a compound that modulates a characteristic associated with one or more condensates comprising a condensate-associated molecule, comprising: (a) contacting the compound with a cellular composition comprising one or more condensates or a cellular composition capable of forming one or more condensates, and (b) determining the characteristic associated with the one or more condensates. A modulation in the characteristic, as compared to a reference, indicates that the compound modulates the characteristic associated with the one or more condensates.

OLIGONUCLEOTIDES FOR INDUCING PATERNAL UBE3A EXPRESSION

The present invention relates to oligonucleotides that are capable of inducing expression of ubiquitin-protein ligase E3A (UBE3A) from the paternal allele in animal or human neurons. The oligonucleotides target the suppressor of the UBE3A paternal allele by hybridization to SNHG14 long non-coding RNA downstream of SNORD109B. The present invention further relates to pharmaceutical compositions and methods for treatment of Angelman syndrome.

CELL-BASED CLOSTRIDAL NEUROTOXIN ASSAYS

The present invention is directed to a method for identifying a gene that regulates clostridial neurotoxin activity, the method comprising: a. providing a sample of human neuronal cells expressing a polypeptide that comprises a C-terminal detectable label, wherein the polypeptide is cleavable by a clostridial neurotoxin; b. altering expression of a target gene of the cells; c. contacting the cells with the clostridial neurotoxin; d. measuring an amount of C-terminal detectable label, thereby quantifying clostridial neurotoxin activity; and e. identifying the target gene as a regulator of clostridial neurotoxin activity when the quantified clostridial neurotoxin activity is different to the quantified clostridial neurotoxin activity when expression of the target gene is unaltered; or f. identifying that the target gene is not a regulator of clostridial neurotoxin activity when the quantified clostridial neurotoxin activity is equivalent to the quantified clostridial neurotoxin activity when ...

ANTI-FAM19A5 ANTIBODIES AND USES THEREOF

The present disclosure provides antibodies that specifically bind to human FAM19A5 and compositions comprising such antibodies. Also provided herein are methods for treating fibrosis or cancer using the anti-FAM19A5 antibodies.

MARKER FOR ANTIDEPRESSANT THERAPY AND METHODS RELATED THERETO

The present invention relates generally to methods for determining the effectiveness of ongoing antidepressant therapy via analysis of the associated of Gs.alpha. with components of the plasma membrane or cytoskeleton of cells from peripheral tissues of the depressed individual as well as to methods involved in screening for effective antidepressant agents via their ability to cause a difference in the association of Gs.alpha. with components of the plasma membrane or cytoskeleton of cells.

METHODS FOR BUILDING GENOMIC NETWORKS AND USES THEREOF

Disclosed are methods, systems, cells and compositions directed to modeling a physiologic or pathologic process in an animal using a set of yeast genes analogous to a set of animal genes and augmenting the physiologic or pathologic process in the animal with predicted gene interactions based on the interactions between the set of yeast genes. Also disclosed are methods of screening for and using therapeutics for neurodegenerative proteinopathies.

METHOD FOR STUDY, DETERMINATION OR EVALUATION BY GENE EXPRESSION ANALYSIS

Disclosed is a method for the study, determination or evaluation of a dis ease condition in a SART-stressed animal or a pharmacological activity of a substance of interest (e.g., an analgetic activity, an autonomic imbalance-a meliorating activity, an anti-stress activity) on the animal at the gene lev el. A disease condition in a SART-stressed animal or a pharmacological activ ity of a substance of interest (particularly, an analgetic activity, an auto nomic imbalance-ameliorating activity, an anti-stress activity) on the anima l can be studied, determined or evaluated at the gene level by administering the substance to the SART-stressed animal and comprehensively analyzing the change in gene expression in a neural tissue of the animal. It becomes poss ible to search for a substance effective for a painful disease, a stress-ind uced disease or autonomic imbalance, to determine/evaluate the effect of the substance, or to analyze a target gene for the substance.

CANCER STEM CELLS AND USES THEREOF

Disclosed are enriched preparations of neuroblastoma tumor initiating cells (NB TICs). The NB TICs are capable of self-renewal, initiating neuroblastoma tumor growth in vivo and are capable of being passaged in high frequency. These NB TICs have chromosomal abnormalities and are capable of giving rise to secondary tumor spheres. Methods are also disclosed for preparing the enriched preparations of NB TICs, such as from neuroblastoma tumor tissue and metastasized bone marrow. Also disclosed are methods of screening candidate substances to identify therapeutic agents for the treatment of neuroblastoma. Methods are also provided for screening a sample for neuroblastoma, as well as for screening a sample to identify the stage of neuroblastoma present. Kits are also provided for selecting appropriate anti-neuroblastoma compounds for a patient, and utilize isolated compositions of the patients' neuroblastoma tumor initiating cells. In this manner, a customized medicinal profile for the patient ...

HIGH THROUGHPUT SCREENING OF AGENTS ON DOPAMINERGIC NEURONS

A method of determining whether an agent is a neuroeffector is disclosed. The method comprises: (a) labeling dopaminergic neurons which are comprised in a mixed population of cells with a fluorescent dopamine analog; (b) measuring a level of fluorescence in the mixed population of cells; (c) exposing the mixed population of cells to the agent; (d) remeasuring a level of fluorescence in the mixed population of cells, wherein a change in the level of fluorescence is indicative of the substance being a neuroeffector.

ANALYSIS OF COMPOUNDS FOR PAIN AND SENSORY DISORDERS

The invention generally relates to optical methods for characterizing the effects of compounds on pain and other sensory phenomena. The effect of compounds on pain and other sensory phenomena may be characterized using dorsal root ganglion (DRG) neurons or sensory neurons expressing optogenetic proteins that allow neural activity to be stimulated and detected optically. The invention provides cell-based optical assays for studying the molecular and cellular bases of pain and sensory phenomena and as platforms to screen and validate drugs, e.g., for pre-clinical trials.

CORTICAL INTERNEURONS AND OTHER NEURONAL CELLS PRODUCED BY THE DIRECTED DIFFERENTIATION OF PLURIPOTENT AND MULTIPOTENT CELLS

Provided are cortical interneurons and other neuronal cells and in vitro methods for producing such cortical interneurons and other neuronal cells by the directed differentiation of stem cells and neuronal progenitor cells. The present disclosure relates to novel methods of in vitro differentiation of stem cells and neural progenitor cells to produce several type neuronal cells and their precursor cells, including cortical interneurons, hypothalamic neurons and pre¬ optic cholinergic neurons. The present disclosue describes the derivation of these cells via inhibiting SMAD and Wnt signaling pathways and activating SHH signaling pathway. The present disclosure relates to the novel discovery that the timing and duration of SHH activation can be harnessed to direct controlled differentiation of neural progenitor cells into either cortical interneurons, hypothalamic neurons or pre-optic cholinergic neurons. The present disclosure also relates to compositions of cortical interneurons, hypothalamic ...

AGENTS THAT REDUCE NEURONAL OVEREXCITATION

The present invention provides methods of identifying candidate agents for treating excitotoxicity-related disorders. The present invention further provides methods for treating excitotoxicity-related disorders.

TREATING MYELIN DISEASES WITH OPTIMIZED CELL PREPARATIONS

The disclosure relates to oligodendrocyte-biased glial progenitor cells and methods of making, isolating, and using such cells.

METHOD OF TREATMENT

Molecular signatures indicate pharmacologic activity.

HIGH MAST2-AFFINITY POLYPEPTIDES AND USES THEREOF

The invention relates to polypeptides containing a cytoplasmic domain ending with a MAST-2 binding domain, from 11 to 13 residues, the first two residues of which are S and W, and the last four residues of which are Q, T, R and L, said polypeptides presenting a high affinity for the PDZ domain of the human MAST2 protein. The invention also relates to polynucleotides, vectors, lentiviral particles, cells as well as compositions comprising the same. The invention is also directed to the use of said polypeptides, polynucleotides, vectors, lentiviral particles, cells and compositions in the treatment and/or prevention of a disease, disorder or condition, which alters the Central Nervous System (CNS) and/or the Peripheral Nervous System (PNS). The invention also concerns molecular signatures of cellular genes to determine the neurosurvival and/or neuroprotection activity of a molecule.

CELLULAR AND ANIMAL MODELS FOR SCREENING THERAPEUTIC AGENTS FOR THE TREATMENT OF ALZHEIMER'S DISEASE

Constructs, models, and methods to assay candidate therapeutic agents to treat Alzheimer's disease are described herein. In an embodiment, a construct includes a polynucleotide encoding a fusion protein of (a) a chaperone protein or monosaccharide transferase (MST) and (b) a fluorescent protein. In an embodiment, the fusion occurs where at least one exon of a polynucleotide encoding a chaperone protein or MST is followed by a polynucleotide encoding a fluorescent protein.

METHODS AND COMPOSITIONS FOR GENE, TUMOR, AND VIRAL INFECTION THERAPY, AND PREVENTION OF PROGRAMMED CELL DEATH (APOPTOSIS)

METHODS AND COMPOSITIONS FOR GENE, TUMOR, AND VIRAL INFECTION THERAPY, AND PREVENTION OF PROGRAMMED CELL DEATH (APOPTOSIS) The present invention relates to methods of treatment of programmed cell death (apoptosis) through the use of the HSV-1 gene .gamma.134.5 or the product of its expression, ICP34.5. The gene and its expression have been demonstrated to be required for HSV-1 neurovirulence, and in particular, to act as an inhibitor of neuronal programmed cell death which allows for viral replication. Use of the gene therapy, or the protein itself, can be expected to result in inhibition of programmed cell death in various neurodegenerative diseases. This invention also relates to novel vectors for gene therapy, including modified herpes virus. Methods are presented for conducting assays for substances capable of mimicing, potentiating or inhibiting the expression of .gamma.134.5 or the activity of ICP34.5. Also, methods are disclosed for the treatment of tumorogenic diseases, including ...

METHODS AND COMPOSITIONS FOR MONITORING CELLULAR PROCESSING OF (.BETA.) -AMYLOID PRECURSOR PROTEIN

... 2118243 9321526 PCTABS00027 Processing of .beta.-amyloid precursor protein (.beta.APP) is monitored by detecting the secretion of a soluble bAPP fragment resulting from cleavage of .beta.APP at the amino-terminus of .beta.-amyloid peptide. in vivo monitoring of secretion of the .beta.APP fragment may be monitored for diagnosis and prognosis of Alzheimer's disease and other .beta.-amyloid-related diseases, while in vitro monitoring of such secretion from cultured cells may be monitored to identify inhibitors of .beta.-amyloid production. The .beta.APP fragment may be detected using antibodies and other specific binding substances which recognize a carboxy-terminal residue on the fragment.

MORPHOGENIC PROTEIN SCREENING METHOD

... 2116560 9305172 PCTABS00020 Disclosed is a method of screening candidate compounds for the ability to modulate the level of morphogenic protein in mammalian system. The method includes determining a parameter indicative of the level of production of a morphogenic in a cell culture known to produce the morphogen, incubating a candidate compound with the culture for a time sufficient to allow the compound to affect the production of the morphogenic protein, and then assaying the culture again to detect a change in the level of morphogenic protein production.

IN VITRO MODELS OF CNS FUNCTION AND DYSFUNCTION

Proliferating or proliferated multipotent neural stem cells and their progeny are used to produce a CNS model system for the study of neural development and function and for determining the CNS effects of novel therapeutic and other biological agents. The neural stem cells are obtained from small amounts of either normal or diseased CNS tissue from pre- and post natal individuals. The invention allows for large amounts of tissue, which may be clonally derived to limit variability, to be generated from a relatively small amount of CNS tissue. The invention describes a CNS model system whereby the differentiated progeny of the neural stem cells include multiple types of CNS cells, including neurons, astrocytes and oligodendrocytes. Screening for the effects of neurological or other biological agents and the analysis of gene expression in the multipotent neural stem cells and in the stem-cell derived progeny of a normal or a diseased donor may be undertaken using this model system.

CNS NEURITE OUTGROWTH MODULATORS, AND COMPOSITIONS, CELLS AND METHODS EMBODYING AND USING SAME

The invention features a method for promoting neural growth in vivo in the mammalian central nervous system by administering a neural cell adhesion molecule which can overcome inhibitory molecular cues found on glial cells and myelin to promote neural growth. Also featured active fragments, cognates, congeners, mimics, analogs, secreting cells and soluble molecules thereof, as well as antibodies thereto, and DNA molecules, vectors and transformed cells capable of expressing them. The invention also includes transgenic mouse lines expressing a neural adhesion molecule in differentiated astrocytes, and cells and tissues derived therefrom. The expression of the neural adhesion molecule enhances neurite outgrowth on central nervous system tissue derived from these transgenic mice. The invention also features methods for enhancing neuronal outgrowth of CNS neurons, for enhancing memory and for increasing synaptic efficacy. Also featured are methods of testing drugs which modulate the effects ...

IDENTIFICATION OF AGENTS THAT PROTECT AGAINST INFLAMMATORY INJURY TO NEURONS

The present invention is directed to methods for identifying agents that inhibit the toxic effects of neurotoxins on neurons from plaque component activated mononuclear phagocytes. In addition, the present invention is directed to methods of identifying agents that inhibit mononuclear phagocyteplaque component complex formation, plaque component activation of mononuclear phagocytes, and plaque component induced neurotoxicity of mononuclear phagocytes. The present invention is also directed to agents and pharmaceutical compositions obtained by the identification methods described herein. Additionally, the present invention describes methods for using tyramine compounds to inhibit the toxic effects of neurotoxins and methods to treat and diagnose neurodegenerative diseases and disorders.

METHOD TO DIAGNOSE AND MONITOR CELLULAR IMMUNE DEFICIENCIES

A method and kit for determining candidates for immunotherapy, for monitoring the effect of immunotherapy and analysis of cell mediated immunity functionality in a patient who needs immunotherapy is provided. The method includes performing two intracutaneous skin tests and reading the skin test after twenty-four hours. One skin test is the administration of a mitogen such as phytohemagglutinin (PHA), concanavalin A (ConA), pokeweed antigen (PWA) and other mitogens as known in the art. The PHA skin test responses reflect the ability of the T-lymphocytes which are present to react to PHA and to release cytokines like IL-2 and induce a monocyte/macrophage infiltration leading to the DTH dermal reaction which is observed in the skin test characteristic of the afferent limb response of the immune system. The NCM (mitogen-stimulated natural cytokine mixture) skin test reflects the ability of preformed T-cell cytokines to induce the monocyte/macrophage accumulation characteristic of the efferent ...

СОЕДИНЕНИЯ БЕНЗИМИДАЗОЛКАРБОКСАМИДА В КАЧЕСТВЕ АГОНИСТОВ РЕЦЕПТОРОВ 5-НТ4

Настоящее изобретение относится к бензимидазолкарбоксамидным агонистам рецептора 5-HT4, являющимся соединением формулы (I) где R1 и X являются такими, как определено в описании, или к их фармацевтически приемлемой соли, или сольвату, или стереоизомеру. Настоящее изобретение также относится к фармацевтическим композициям, содержащим такие соединения, к способам применения таких соединений для лечения заболеваний, связанных с активностью рецептора 5-HT4, и к способам и промежуточным продуктам, полезным для получения таких соединений. Настоящее изобретение дополнительно относится к кристаллическим формам соединений формулы (I).

СОЕДИНЕНИЯ БЕНЗИМИДАЗОЛКАРБОКСАМИДА В КАЧЕСТВЕ АГОНИСТОВ РЕЦЕПТОРОВ 5-НТ4

Настоящее изобретение относится к бензимидазолкарбоксамидным агонистам рецептора 5-НТ4, являющимся соединениями формулы (I) где R1 и Х являются такими, как определено в описании, или к их фармацевтически приемлемой соли, или сольвату, или стереоизомеру. Настоящее изобретение также относится к фармацевтическим композициям, содержащим такие соединения, к способам применения таких соединений для лечения заболеваний, связанных с активностью рецептора 5-НТ4, и к способам и промежуточным продуктам, полезным для получения таких соединений. Настоящее изобретение дополнительно относится к кристаллическим формам соединений формулы (I).

МОДУЛЯТОРЫ ОТРАЩИВАНИЯ НЕЙРИТОВ ЦНС, А ТАКЖЕ СОСТАВЫ, КЛЕТКИ И СПОСОБЫ ИХ ВОПЛОЩЕНИЯ И ИСПОЛЬЗОВАНИЯ

МИКРООРГАНИЗМЫ ИЛИ ИХ ФРАКЦИИ, ОБЕСПЕЧИВАЮЩИЕ АКТИВАЦИЮ КЛЕТОЧНОГО ИММУНИТЕТА ПРОТИВ УГЛЕВОДОВ

Настоящее изобретение относится к профилактике и лечению желудочно-кишечных расстройств и рака. В частности, изобретение относится к профилактике и лечению карцином, которые являются Core-1-положительными. Это изобретение касается кориотиков и способа их получения, а также способа предотвращения и лечения Core-1-положительных расстройств, используя эти кориотики.

ВЫДЕЛЕННАЯ НУКЛЕИНОВАЯ КИСЛОТА, КОДИРУЮЩАЯ ПОЛИПЕПТИД Sp35, ПОЛИПЕПТИД Sp35 И СПОСОБЫ ПРИМЕНЕНИЯ НУКЛЕИНОВОЙ КИСЛОТЫ И ПОЛИПЕПТИДА

Изобретение относится к полипептидам Sp35 и их белкам слияния, антителам против Sp35 и их антигенсвязывающим фрагментам, а также к кодирующим указанные белки нуклеиновым кислотам. Изобретение также относится к композициям, содержащим такие антитела против Sp35, их антигенсвязывающие фрагменты, полипептиды Sp35 и их белки слияния, и к способам получения и применения указанных белков.

ВЫДЕЛЕННЫЙ ПОЛИНУКЛЕОТИД (ВАРИАНТЫ), СОДЕРЖАЩИЙ ЕГО ВЕКТОР И КЛЕТКА-ХОЗЯИН, КОДИРУЕМЫЙ ИМ ПОЛИПЕПТИД РЕЦЕПТОРА NOGO, УМЕНЬШАЮЩИЙ ОПОСРЕДОВАННОЕ ЭТИМ РЕЦЕПТОРОМ ИНГИБИРОВАНИЕ РОСТА АКСОНОВ (ВАРИАНТЫ), ВЫДЕЛЕННОЕ АНТИТЕЛО И ФАРМАЦЕВТИЧЕСКАЯ КОМПОЗИЦИЯ НА ИХ ОСНОВЕ

Описаны белки рецептора NOGO и пептиды, которые блокируют опосредованное NOGO ингибирование удлинение аксонов, а также выделенные полинуклеотиды, их кодирующие. Композиции по изобретению могут применяться при лечении черепной или церебральной травмы, повреждения спинного мозга, инсульта или демиелинизирующего заболевания.

Methods of treating neurodegenerative disorders and diseases

This invention is directed to a novel method of treating neurodegenerative disorders and diseases. Another, related aspect of this invention is directed to a screening method of identifying compounds that can be used to treat neurodegenerative disorders and diseases. The foregoing aspects of the invention particularly relate to neurodegenerative disorders and diseases have degeneration of neuronal axons as part of their pathologies. The method of treatment involves administering a pharmaceutical formulation that comprises a compound or mixture of compounds that inhibits one or more intracellular signaling mechanism that regulate axon degeneration or growth cone collapse. The screening method aspect of the invention identifies test compounds that can be used for the treatment or prevention of neurodegenerative disorders based on the test compound's ability to inhibit axon degeneration or growth cone collapse.

Multipotent neural stem cells

An isolated multipotent neural stem cells has a phenotype identified by expression of the protein β-tubulin IV and Olig2 and the absence of the proteins NG2, PLP, and GFAP.

Novel Target for Regulating Multiple Sclerosis

Methods are provided for decreasing demyelinating inflammatory disease in a subject by inhibiting the activity of chemokine-like receptor 1 (CMKLR1). Methods are also provided for screening for agents that find use in treating demyelinating inflammatory disease in a subject.

Pharmaceutical composition containing nuclear factor involved in proliferation and differentiation of central neuronal cells

With an aim to provide a novel factor inducing proliferation of neural stem cells and differentiation of these cells into nerve cells, a pharmaceutical composition comprising 1) CRBN, 2) a nucleic acid encoding CRBN, or 3) a stem cell or a neural progenitor cell in which CRBN is expressed, a method including administering the pharmaceutical composition to a non-human animal and inducing proliferation of neural stem cells or neural progenitor cells of the non-human animal and differentiation of these cells into nerve cells, and a method for screening for a therapeutic drug for a disease of cerebral cortex or a surgical injury of cerebral cortex, using CRBN, are provided.

Method and Pharmaceutical Composition for Use in the Treatment of Neurodegenerative Disorders

The invention relates to compounds which activate the BASIGIN signaling pathway, preferably agonists of BASIGIN, for the treatment of neurodegenerative disorders.

Modulation of Synaptic Maintenance

C1q is shown to be expressed in neurons, where it acts as a signal for synapse elimination. Methods are provided for protecting or treating an individual suffering from adverse effects of synapse loss. These findings have broad implications for a variety of clinical conditions, including Alzheimer's disease.

Technologies, Methods, and Products of Small Molecule Directed Tissue and Organ Regeneration from Human Pluripotent Stem Cells

Pluripotent human embryonic stem cells (hESCs) hold great potential for restoring tissue and organ function, which has been hindered by inefficiency and instability of generating desired cell types through multi-lineage differentiation. This instant invention is based on the discovery that pluripotent hESCs maintained under defined culture conditions can be uniformly converted into a specific lineage by small molecule induction. Retinoic acid induces specification of neuroectoderm direct from the pluripotent state of hESCs and triggers progression to neuronal progenitors and neurons efficiently. Similarly, nicotinamide induces specification of cardiomesoderm direct from the pluripotent state of hESCs and triggers progression to cardiac precursors and cardiomyocytes efficiently. This technology provides a large supply of clinically-suitable human neuronal or cardiac therapeutic products for CNS or myocardium repair. This invention enables well-controlled efficient induction of pluripotent hESCs exclusively to a specific clinically-relevant lineage for tissue and organ engineering and regeneration, cell-based therapy, and drug discovery.

Methods of controlling axon or dendrite development of neuronal cells

One aspect of the present invention relates to a method of controlling axon or dendrite development in a neuronal cell population. This method involves providing a neuronal cell population and contacting the neuronal cell population with a modulator of R-type Ca 2+ channel expression or activity in controlling the neuronal cell population to induce either axon or dendrite development. Another aspect of the present invention relates to a method of treating neuronal injury in a subject. This method involves selecting a subject with neuronal injury mediated by R-type Ca 2+ channel expression or activity and administering to the selected subject an inhibitor of R-type Ca 2+ channel expression or activity to induce neuronal axon development under conditions effective to treat the neuronal injury in the subject. Yet another aspect of the present invention relates to a method of screening for agents that modulate R-type Ca 2+ channel expression or activity.

Therapeutic Applications Targeting SARM1

The present disclosure provides methods for reducing axonal and/or synaptic degradation in neurons by modulating sterile α/Armadillo/Toll-Interleukin receptor homology domain protein (SARM) activity and/or expression.

Methods and compositions for treating neurological disorders

Provided herein are methods and compositions useful in treating neurological disorders.

Model and Methods for Identifying Points of Action in Electrically Active Cells

The invention provides a model for generating predicted action potentials of an electrically active cell. The disclosed model includes three operatively coupled submodels. A first submodel contains Hodgkin-Huxley elements generating action potentials based on electrical equivalent circuits. A second submodel is based on reaction kinetics of cell metabolism and is operatively coupled with the first submodel. A third submodel is based on Boolean dynamics representing signaling and associated cellular processes and is operatively coupled with the first and second submodels. The invention includes storing a library of calculated action potentials and associated cellular parameters generated by the model, applying a stimulus to the electrically active cell in vitro so that the cell generates an action potential; and comparing the cell-generated action potential with those stored in the library, wherein a match is predictive of the cellular point of action of the applied stimulus according to the parameters stored.

Fluorogenic ph sensitive dyes and their method of use

A new class of pH sensitive fluorescent dyes and assays relating thereto are described. The dyes and assays are particularly suited for biological applications including phagocytosis and monitoring intracellular processes. The pH sensitive fluorescent dyes of the present invention include compounds of Formula I: wherein the variables are described throughout the application.

Formation of neuromuscular junctions in a defined system

A method for forming neuromuscular junctions includes forming functional neuromuscular junctions between motoneurons and muscle cells by co-culturing one or more human motoneurons and one or more human muscle cells in a substantially serum-free medium. A synthetic mammalian neuromuscular junction includes a human motoneuron functionally linked to a human muscle cell in a substantially serum-free medium. An artificial substrate may be used to support the one or more neuromuscular junctions.

Wound Healing Metakaryotic Stem Cells and Methods of Use Thereof

The invention provides methods of identifying wound healing metakaryotic stem cells, identifying molecules to modulate proliferation and/or migration of metakaryotic stem cells and molecules to treat wound healing disorders, such as blood vessel wound healing disorders, including restenosis. The invention also provides methods of diagnosing and treating wound healing disorders, such as blood vessel wound healing disorders and also of treating wounds by the use of metakaryotic stem cell transplant(s).

Microfluidic device for generating neural cells to simulate post-stroke conditions

This application provides devices for modeling ischemic stroke conditions. The devices can be used to culture neurons and to subject a first population of the neurons to low-oxygen conditions and a second population of neurons to normoxic conditions. The neurons are cultured on a porous barrier, and on the other side of the barrier run one or more fluid-filled channels. By flowing fluid with different oxygen levels through the channels, one can deliver desired oxygen concentrations to the cells nearest those channels.

GREEN TEA POLYPHENOL ALPHA SECRETASE ENHANCERS AND METHODS OF USE

The subject invention concerns materials and methods for treating or preventing a neurodegenerative condition or disease associated with β-amyloid peptide deposition in neural tissue in a person or animal by administering a therapeutically effective amount of a polyphenol, or an analog, isomer, metabolite, or prodrug thereof, that increases expression or activity of a protein that exhibits α-secretase activity. The subject invention also provides methods to increase α-secretase expression and/or activity in cells by administering polyphenol flavonoids like (−)-epigallocatechin--gallate (EGCG) and epicatechin (EC), two polyphenols derived from green tea and other plants and that can be produced synthetically. Furthermore, there are provided methods to decrease or inhibit the production of Aβor Aβby administering the EGCG and EC compounds, their analogs, metabolites, and prodrugs. 1. A method for:i) treating or preventing a neurodegenerative disease or condition associated with β-amyloid peptide deposition in neural tissue, said method comprising administering to a person or animal in need thereof an effective amount of a polyphenol, or an analog, isomer, metabolite, or prodrug thereof, that increases expression or activity of a protein having α-secretase enzymatic activity; orii) decreasing or inhibiting deposition of β-amyloid peptide in neural tissue, said method comprising contacting said neural tissue with an effective amount of a polyphenol, or an analog, isomer, metabolite, or prodrug thereof, that increases expression or activity of a protein having α-secretase enzymatic activity; oriii) decreasing levels of a β-amyloid peptide produced by a cell, said method comprising contacting a cell with an effective amount of a polyphenol, or an analog, isomer, metabolite, or prodrug thereof, that increases expression or activity of a protein having α-secretase enzymatic activity.2. The method according to claim 1 , wherein said polyphenol is epigallocatechin-3-gallate ( ...

METHOD FOR DETERMINING ACTIVATORS OF EXCITATORY SYNAPSE FORMATION

The invention provides methods of screening a compound that can increase spine/excitatory synapse formation and/or numbers. The compound is identified by contacting Ephexin5 with a test compound and selecting the compounds that inhibit Rho GEF activity of Ephexin5. Additionally, the invention also provides methods for increasing spine/excitatory synapse formation and/or numbers by contacting a neuron with an Ephexin5 inhibitor. 1. A method for determining activators of spine/excitatory synapse formation , comprising contacting Ephexin5 with a test compound and selecting the compound that inhibits Ephexin5 RhoA GEF activity.2. The method of claim 1 , wherein Ephexin5 RhoA GEF activity is determined by assaying RhoA activation and selecting the test compound that decreases RhoA activation relative to a control.3. The method of claim 2 , wherein RhoA activation is determined by binding of GTP to RhoA or by assaying the GTPase activity of RhoA or by assaying binding of a protein to RhoA claim 2 , which protein preferentially binds to activated RhoA.4. (canceled)5. (canceled)6. (canceled)7. The method of claim 3 , wherein the protein comprises a Rhotekin-Binding Domain (RBD).8. (canceled)9. (canceled)10. The method of claim 1 , wherein the Ephexin5 comprises an amino acid sequence or a conservative variant thereof encoded by a polynucleotide sequence selected from the group consisting of NCBI Sequence Reference Nos. and/or identifier Rho guanine nucleotide exchange factor (GEF) 15 claim 1 , ARGEF15 claim 1 , KIAA0915 claim 1 , F1113791 claim 1 , Vsm-RhoGEF claim 1 , Rho guanine exchange factor (GEF) 15 claim 1 , MGC44868 claim 1 , rho guanine nucleotide exchange factor 15 claim 1 , 155901 claim 1 , 228992 claim 1 , ENSG000001988447 claim 1 , O949893 claim 1 , GC17P008591 claim 1 , GC17P009307 claim 1 , GC17P008156 claim 1 , GC17P008414 claim 1 , MGC102247 claim 1 , MGC144141 claim 1 , MGC144142 claim 1 , D130071N09 claim 1 , D530030K12Rik claim 1 , RP23-396M19.6 claim 1 ...

BENZIMIDAZOLE-CARBOXAMIDE COMPOUNDS AS 5-HT4 RECEPTOR AGONISTS

The invention relates to benzimidazole-carboxamide 5-HTreceptor agonist compounds of formula (I) 122-. (canceled)24. The method of wherein in the compound of formula (I) claim 23 ,{'sup': '1', 'Ris isopropyl or tert-butyl; and'} [{'sup': 2', '2, '(a) —C(O)ORwherein Ris methyl or phenyl;'}, {'sup': 3', '3, 'sub': '3', '(b) —C(O)Rwherein Ris phenyl, optionally substituted with 1 or 2 substituents selected from methyl, chloro, fluoro, and —CF; furan-2-yl; or thiophen-2-yl; and'}, {'sup': 4', '5', '4', '5, '(c) —C(O)NRRwherein Ris hydrogen and Ris phenyl optionally substituted with 1 fluoro or chloro.'}], 'X is selected from This application claims the benefit of U.S. Provisional Applications Nos. 60/684,466 and 60/684,478, filed on May 25, 2005, and 60/748,415, filed on Dec. 8, 2005, the disclosures of which are incorporated herein by reference in their entirety.1. Field of the InventionThe invention is directed to benzimidazole-carboxamide compounds which are useful as 5-HTreceptor agonists. The invention is also directed to pharmaceutical compositions comprising such compounds, methods of using such compounds for treating or preventing medical conditions mediated by 5-HTreceptor activity, and processes and intermediates useful for preparing such compounds.2. State of the ArtSerotonin (5-hydroxytryptamine, 5-HT) is a neurotransmitter that is widely distributed throughout the body, both in the central nervous system and in peripheral systems. At least seven subtypes of serotonin receptors have been identified and the interaction of serotonin with these different receptors is linked to a wide variety of physiological functions. There has been, therefore, substantial interest in developing therapeutic agents that target specific 5-HT receptor subtypes.In particular, characterization of 5-HTreceptors and identification of pharmaceutical agents that interact with them has been the focus of significant recent activity. (See, for example, the review by Langlois and ...

NOGO Receptor Binding Protein

The invention provides Sp35 polypeptides and fusion proteins thereof, Sp35 antibodies and antigen-binding fragments thereof and nucleic acids encoding the same. The invention also provides compositions comprising, and methods for making and using, such Sp35 antibodies, antigen-binding fragments thereof, Sp35 polypeptides and fusion proteins thereof. 157-. (canceled)58. A method of identifying an Sp35 antagonist capable of decreasing inhibition of axonal growth of a central nervous system (CNS) neuron comprising contacting a CNS neuron with a compound and measuring axonal growth of the CNS neuron , wherein increased axonal growth compared to a control CNS neuron indicates the compound is capable of decreasing inhibition of axonal growth of a CNS neuron.59. The method of claim 58 , wherein the Sp35 antagonist binds to Sp35.60. The method of claim 58 , wherein the CNS neuron is in vitro.61. The method of claim 58 , wherein the CNS neuron is in vivo.62. The method of claim 58 , wherein the CNS neuron is a cerebellar granule neuron.63. The method of claim 58 , wherein axonal growth is measured using βIII tubulin staining.64. The method of claim 58 , wherein the Sp35 antagonist inactivates RhoA in the neuron.65. The method of claim 58 , wherein the Sp35 antagonist decreases CA flux in the neuron.66. The method of claim 58 , wherein the Sp35 antagonist causes a decrease in inhibition of axonal growth of a CNS neuron comparable to the decrease that occurs in the presence of a soluble Sp35 polypeptide comprising amino acids 34-532 of SEQ ID NO:2.67. A method of identifying an Sp35 antagonist capable of promoting neuronal cell survival comprising contacting a neuron with a compound and measuring neuronal cell survival claim 58 , wherein increased neuronal survival compared to a control neuron indicates the compound is an Sp35 antagonist capable of promoting neuronal cell survival.68. The method of claim 67 , wherein the Sp35 antagonist binds to Sp35.69. The method of claim 67 ...

Cell-Based Models of Neurodegenerative Disease

The present invention relates to a cell-based model useful for identifying molecules that modify intracellular pathways of α-synuclein and tau aggregation and degradation. 1. A cell culture model for alpha synuclein inclusion formation , said model comprising a cell population and exogenous alpha synuclein fibrils.2. The cell culture model of claim 1 , wherein said cell population comprises a neuronal cell.3. The cell culture model of claim 1 , wherein said cell population comprises a non-genetically modified cell.4. The cell culture model of claim 1 , wherein said cell population comprises a cell engineered to express a nucleic acid encoding alpha synuclein.5. The cell culture model of claim 4 , wherein said cell further comprises a nucleic acid encoding a detectable protein.6. The cell culture model of claim 1 , comprising a culture medium comprising exogenous alpha synuclein fibrils.7. The cell culture model of claim 1 , further comprising wheat germ agglutinin (WGA).8. The cell culture model of claim 1 , wherein said exogenous alpha synuclein fibrils are derived from a mammalian alpha synuclein.9. The cell culture model of claim 8 , wherein said mammalian alpha synuclein is selected from the group consisting of mouse claim 8 , rat claim 8 , primate claim 8 , and human.10. The cell culture model of claim 1 , wherein said exogenous alpha synuclein fibrils is derived from a human alpha synuclein or a fragment thereof selected from the group consisting of full-length alpha synuclein (α-syn) claim 1 , α-syn-1-120 claim 1 , α-syn-1-89 claim 1 , α-syn-58-140 claim 1 , α-syn-61-95 claim 1 , and any combination thereof.11. A cell culture model for tau inclusion formation claim 1 , said model comprising a cell engineered to express a nucleic acid encoding tau.12. The cell culture model of claim 11 , wherein said tau has a P301L mutation.13. The cell culture model of claim 11 , wherein said cell is a neuronal cell.14. The cell culture model of claim 11 , wherein said cell ...

Alzheimer's Disease Cellular Model for Diagnostic and Therapeutic Development

Stem-cell derived human neuronal models that mimic human Alzheimer's disease, including hereditary and sporadic Alzheimer's disease, comprising neural stem cells derived from human induced pluripotent stem cells. Also provided are purified human neurons developed from the neural stem cells that carry genomes from the Alzheimer's disease patients. The human neuronal models are neuronal models for hereditary and sporadic Alzheimer's disease, and are suitable for measurement of key behaviors of the Alzheimer's disease, providing further diagnostic tools for the development of sporadic Alzheimer's disease, and assisting in drug testing for the therapeutic treatment of Alzheimer's disease. 1. A human neuronal model comprising human neural cells derived from human induced pluripotent stem cells (iPSCs) from a patient with Alzheimer's disease.2. The human neuronal model of claim 1 , wherein said Alzheimer's disease is sporadic Alzheimer's disease.3. The human neuronal model of claim 1 , wherein said Alzheimer's disease is hereditary Alzheimer's disease.4. The human neuronal model of claim 1 , further comprising human neurons developed from said human neural cells.5. The human neuronal model of claim 4 , wherein said human neurons present key behaviors of Alzheimer's disease.6. The human neuronal model of claim 5 , wherein said key behaviors are measurements of proteolytic processing of one or more amyloid precursor protein claim 5 , phosphorylation of tau protein claim 5 , activation of GSk3 kinase claim 5 , measurement of synaptic phenotype claim 5 , autophagy claim 5 , or other disease behaviors.7. A method of making a human neuronal model of claim 1 , comprising:a) isolating fibroblasts from a patient with Alzheimer's disease;b) reprogramming said fibroblasts to induced pluripotent stem cells (iPSCs);c) further differentiating said iPSCs into cultures containing neural rosettes;d) purifying neural precursor cells (NPCs);e) further differentiating purified NPCs into ...

METHODS AND COMPOSITIONS FOR TREATING NEUROPATHIES

Methods of treating or preventing axonal degradation in neuropathic diseases and neurological disorders in mammals are disclosed. The methods can comprise administering to the mammal an effective amount of an agent that acts at least in part by increasing sirtuin AMPK activity, LKB activity and/or CaMKKβ activity in diseased and/or injured neurons. The methods can also comprise administering to the mammal an effective amount of an agent that acts by increasing NAD activity in diseased and/or injured neurons, alone or in combination with agents that act by other mechanisms. Also disclosed are methods of screening agents for treating a neuropathies and recombinant vectors for treating or preventing such neuropathies. 1. A method of promoting axonal growth in a mammal in need thereof , the method comprising administering to the mammal an agent in an amount effective for promoting axonal growth by increasing at least one of AMP activated kinase (AMPK) activity , LKB1 activity and CaMKKβ activity in at least one of diseased neurons , injured neurons and supporting cells.2. A method according to claim 1 , wherein the agent is a stilbene claim 1 , a chalcone claim 1 , a flavone claim 1 , an isoflavanone claim 1 , a flavanone or a catechin.3. A method according to claim 2 , wherein the stilbene is selected from the group consisting of resveratrol claim 2 , piceatannol claim 2 , deoxyrhaponfin claim 2 , trans-stilbene and rhapontin.4. A method according to claim 2 , wherein the chalcone is selected from the group consisting of burtein claim 2 , isoliquiritigenin and 3 claim 2 ,4 claim 2 ,2′ claim 2 ,4′ claim 2 ,6′-pentahydroxychalcone.5. A method according to claim 2 , wherein the flavone is selected from the group consisting of fisetin claim 2 , 5 claim 2 ,7 claim 2 ,3′ claim 2 ,4′ claim 2 ,5′-pentahydroxyflavone claim 2 , luteolin claim 2 , 3 claim 2 ,6 claim 2 ,3′ claim 2 ,4′-tetrahydroxyflavone claim 2 , quercetin claim 2 , 7 claim 2 ,3′ claim 2 ,4′ claim 2 ,5′- ...

AMYLOIDOSIS TARGET USEFUL IN METHODS OF TREATMENT AND FOR SCREENING OF COMPOUNDS

A method involves screening a candidate compound for activity in the treatment of a condition associated with formation of amyloid protein fibrils in a mammal, such as Alzheimer's disease. It is determined whether the trimer/monomer ratio of a chaperone protein is decreased in the presence of the candidate compound. The chaperone protein is or has a high identity to the Brichos domains of Bri2, Bri3 or proSP-C from human. Monomers of the chaperone proteins and/or compounds that promote formation of these monomers are useful for medical treatment of the condition. 1. A method of screening one or more candidate compound(s) for activity in the treatment of a condition associated with formation of amyloid protein fibrils in a mammal , comprising determining whether the trimer/monomer ratio of a chaperone protein is decreased in the presence of said one or more candidate compound(s) , wherein the chaperone protein is comprising more than or equal to 80 amino acid residues and is comprising an amino acid sequence having at least 70% identity to an amino acid sequence selected from the group consisting of residues 90-243 of Bri2 from human (SEQ ID NO: 2) , the Brichos domain of Bri2 from human (SEQ ID NO: 3) , residues 97-242 of Bri3 from human (SEQ ID NO: 5) , the Brichos domain of Bri3 from human (SEQ ID NO: 6) , CTproSP-C from human (SEQ ID NO: 8) , and the Brichos domain of CTproSP-C from human (SEQ ID NO: 9).2. A method according to claim 1 , wherein the chaperone protein comprises less than or equal to 200 amino acid residues.3. A method according to claim 2 , wherein the chaperone protein comprises less than or equal to 150 amino acid residues.4. A method according to claim 1 , wherein the chaperone protein is selected from the group consisting of residues 90-243 of Bri2 from human (SEQ ID NO: 2) claim 1 , residues 97-242 of Bri3 from human (SEQ ID NO: 5) claim 1 , and CTproSP-C from human (SEQ ID NO: 8).5. A method according to claim 1 , wherein the chaperone ...

MICROFLUIDIC BIOLOGICAL BARRIER MODEL AND ASSOCIATED METHOD

A biological barrier model is disclosed. In some embodiments the barrier may be configured to model the blood brain barrier. The model may include a membrane having one or more cell cultures disposed thereon. The cells cultures may be grown in the presence of shear stress induced by flow through the device in some embodiments. The size of the barrier, as well as the distance to electrodes and other sensors, may be in the microscale range. Further, in some embodiments the model may comprise an array of parallel channels and membranes. 1. A biological barrier model comprising:a first microfluidic channel;a second microfluidic channel; anda semipermeable membrane disposed at an interface between the first microfluidic channel and the second microfluidic channel.2. The biological barrier model of claim 1 , wherein both the first microfluidic channel and the second microfluidic channel are about 200 μm high.3. The biological barrier model of claim 2 , wherein the first microfluidic channel is about 2 mm wide at the interface and the second microfluidic channel is about 5 mm wide at the interface.4. The biological barrier model of claim 2 , wherein the first microfluidic channel is about 2 mm wide at the interface and the second microfluidic channel is about 2 mm wide at the interface.5. The biological barrier model of claim 1 , wherein the first microfluidic channel is configured to promote uniform shear stress on a first layer of cells cultured on a first surface of the semipermeable membrane claim 1 , and wherein the first microfluidic channel has an aspect ratio of at least 10:1.6. The biological barrier model of claim 1 , further comprising:a first culture of cells on a first surface of the semipermeable membrane, wherein the first surface is in fluid communication with the first microfluidic channel; anda second culture of cells on a second surface of the semipermeable membrane, wherein the second surface in fluid communication with the second microfluidic channel.7 ...

Therapeutic Applications Targeting SARM1

The present disclosure provides methods for reducing axonal and/or synaptic degradation in neurons by modulating sterile α/Armadillo/Toll-Interleukin receptor homology domain protein (SARM) activity and/or expression. 1. A method reducing axonal and/or synaptic degradation in a neuron , the method comprising:selecting a neuron with or at risk for axonal and/or synaptic degradation; andcontacting the neuron with an effective amount of a composition that inhibits sterile α/Armadillo/Toll-Interleukin receptor homology domain protein (SARM) activity and/or expression for a time sufficient to inhibit SARM activity and/or expression, thereby reducing axonal and/or synaptic degradation in the neuron.2. A method for reducing axonal and/or synaptic degradation in a subject with or at risk for developing axonal and/or synaptic degradation , the method comprising:selecting a subject with or at risk for developing axonal and/or synaptic degradation; andtreating the subject with an effective amount of a composition that inhibits SARM activity and/or expression for a time sufficient to inhibit SARM activity and/or expression, thereby reducing axonal and/or synaptic degradation in the subject.3. The method of claim 2 , wherein the subject has or is at risk of neurodegenerative disease.4. The method of or claim 2 , wherein the axonal and/or synaptic degradation is in the central nervous system (CNS) and/or the peripheral nervous system (PNS).5. The method of claim 4 , wherein the axonal and/or synaptic degradation is in the PNS.6. The method of claim 5 , wherein the subject has diabetes.7. The method of claim 6 , wherein the subject has diabetic neuropathy.8. The method of claim 5 , wherein the subject is scheduled to receive chemotherapy.9. The method of claim 5 , wherein the subject is receiving chemotherapy or has received chemotherapy.10. A method for identifying a compound that inhibits SARM activity and/or expression claim 5 , the method comprising:providing SARM;contacting ...

identification of compounds that disperse tdp-43 inclusions

Herein, methods of modulating inclusion formation and stress granules in cells are described. The methods comprise contacting a cell with an inclusion inhibitor. Methods for screening for modulators of TDP-43 aggregation are also described.

Control and Characterization of Psychotic States

Provided herein are methods of inducing psychosis in animals using light-responsive opsins and methods of identifying or screening compounds that may be useful in treating psychosis. 1. A non-human animal comprising a light-responsive opsin expressed on the cell membrane of a subset of layer V pyramidal neurons in the prefrontal cortex , wherein light activation of the opsin induces depolarization of the membrane and induces psychosis of the animal.2. The animal of claim 1 , wherein the subset of layer V pyramidal neurons have a single large apical dendrite.3. The animal of claim 1 , wherein the opsin is selected from the group consisting of ChR2 claim 1 , VChR1 claim 1 , and DChR.4. (canceled)5. A prefrontal cortex tissue slice comprising a subset of layer V pyramidal neurons claim 1 , wherein a light-responsive opsin is expressed on the cell membrane of the apical dendrites in layer V pyramidal neurons claim 1 , and wherein light activation of the opsin induces depolarization of the membrane.6. The prefrontal cortex tissue slice of claim 5 , wherein the subset of layer V pyramidal neurons have a single large apical dendrite.7. The prefrontal cortex tissue slice of claim 5 , wherein the opsin is selected from the group consisting of ChR2 claim 5 , VChR1 claim 5 , and DChR.8. The prefrontal cortex tissue of claim 5 , wherein the opsin is selected from the group consisting of SFO claim 5 , SSFO claim 5 , C1V1 claim 5 , C1V1-E122T claim 5 , C1V1-E162T claim 5 , and C1V1-E122T/E162T.9. A method of inducing psychosis in a non-human animal claim 5 , comprising activating a light-responsive opsin by light claim 5 , wherein the light-responsive opsin is expressed on the cell membrane of a subset of layer V pyramidal neurons in the prefrontal cortex in the animal claim 5 , and wherein the light activation of the opsin induces depolarization of the cell membrane.10. The method of claim 9 , wherein the subset of layer V pyramidal neurons have a single large apical dendrite.11 ...

TDP-43-STORING CELL MODEL

Disclosed is a transformed cell (a cell model) which can form a cytoplasmic inclusion body derived from TAR DNA-binding protein of 43 kDa (TDP-43) that is found in the brain of a patient suffering from a neurodegenerative disease such as FTLD and ALS. The transformed cell is characterized by having, introduced therein, a promoter capable of functioning in a host cell and a mutant TDP-43 gene. 1. A transformed cell having a mutant TDP-43 gene operably linked to a promoter introduced therein , wherein the mutant TDP-43 gene encodes any one of the following proteins (a) to (d):(a) a protein having an amino acid sequence consisting of amino acids 162-414 of the amino acid sequence of wild-type TDP-43;(b) a protein having an amino acid sequence consisting of amino acids 218-414 of the amino acid sequence of wild-type TDP-43;(c) a protein having an amino acid sequence consisting of amino acids 1-161 of the amino acid sequence of wild-type TDP-43; and(d) a protein that has an amino acid sequence having one to ten amino acids deleted from, substituted in or added to the amino acid sequence (a), (b) or (c) and that has an activity of forming an intracellular inclusion.2. The cell according to claim 1 , wherein the mutant TDP-43 has no CFTR exon 9 skipping activity.3. The cell according to claim 1 , which is a transformed mammal cell.4. The cell according to claim 3 , wherein the mammal cell is a central nervous system cell claim 3 , a peripheral nervous system cell or a neuroblast.5. A method for screening a therapeutic drug for a neurodegenerative disease claim 1 , comprising the steps of: causing the cell according to to make contact with a candidate substance to measure a cellular activity of the cell; and using the obtained measurement result as an indicator.6. The method according to claim 5 , wherein the cellular activity is at least one selected from the group consisting of proliferation capacity claim 5 , viability claim 5 , and the rate claim 5 , number and size of ...

Cell lines expressing inserted secretable reporter genes at multiple stages of differentiation

A composition of matter comprises one or more cell lines configured to inducibly differentiate to at least a first stage of differentiation and a second, subsequent stage of differentiation. Each of the one or more cell lines are genetically edited to express one or more first stage inserted secretable reporter genes placed under control of promoters for genes canonically expressed during the first stage of differentiation. The cell lines are further genetically edited to express one or more second stage inserted secretable reporter genes placed under control of promoters for genes canonically expressed during the second stage of differentiation, but not during the first stage of differentiation, wherein the one or more second stage inserted secretable reporter genes are different than the one or more first stage inserted secretable reporter genes.

LIGHT-ACTIVATED CHIMERIC OPSINS AND METHODS OF USING THE SAME

Provided herein are compositions comprising light-activated chimeric proteins expressed on plasma membranes and methods of using the same to selectively depolarize excitatory or inhibitory neurons. 150.-. (canceled)51. A mammalian cell comprising a polynucleotide comprising a nucleotide sequence encoding a light-responsive chimeric polypeptide comprising an amino acid sequence having at least 95% amino acid sequence identity to the amino acid sequence set forth in SEQ ID NO:1 , wherein the light-responsive chimeric polypeptide is present in the cell membrane.52. The mammalian cell of claim 51 , wherein the light-responsive chimeric polypeptide comprises a C-terminal trafficking signal.53. The mammalian cell of claim 52 , wherein the trafficking signal comprises the amino acid sequence KSRITSEGEYIPLDQIDINV (SEQ ID NO:15).54. The mammalian cell of claim 51 , wherein the nucleotide sequence is operably linked to a promoter.55. The mammalian cell of claim 51 , wherein the light-responsive chimeric polypeptide comprises a Glu to Thr amino acid substitution at position 122 relative to the amino acid sequence set forth in SEQ ID NO:l.56. The mammalian cell of claim 51 , wherein the light-responsive chimeric polypeptide comprises a Glu to Thr amino acid substitution at position 162 relative to the amino acid sequence set forth in SEQ ID NO:l.57. The mammalian cell of claim 51 , wherein the light-responsive chimeric polypeptide comprises a Glu to Thr amino acid substitution at position 122 and a Glu to Thr amino acid substitution at position 162 relative to the amino acid sequence set forth in SEQ ID NO:l.58. The mammalian cell of claim 51 , wherein the cell is a neuronal cell claim 51 , a muscle cell claim 51 , or a stem cell.59. The mammalian cell of claim 51 , wherein the cell is an excitatory neuronal cell or an inhibitory neuronal cell.60. The mammalian cell of claim 51 , wherein the cell further comprises a second light-activated polypeptide present in the cell ...

CONVERSION OF NON-NEURONAL CELLS INTO NEURONS

The subject invention concerns materials and methods for conversion of non-neuronal cells into neurons. The subject invention also concerns methods for screening drugs and other compounds for activity in treating Alzheimer's disease using neuronal cells produced using the subject invention. The subject invention also concerns neuronal cells that have been produced using the methods of the invention. The subject invention also concerns methods for evaluating therapeutic treatment for efficacy in a person or animal having Alzheimer's disease or other neurodegenerative diseases. The subject invention also concerns methods of treating Alzheimer's disease or other neurodegenerative diseases or conditions in a person or animal. 1. A neuronal cell produced by conversion of a non-neuronal cell to a neuronal cell by:i) incorporating into said non-neuronal cell a polynucleotide of polypyrimidine-tract-binding (PTB) protein short hairpin RNA (shRNA) and a polynucleotide encoding a selectable marker;ii) selecting for a cell expressing said selectable marker;iii) culturing said selected cell in a neural induction medium comprising a ROCK inhibitor for a sufficient period of time to induce neuronal development, whereby said selected cell is converted into a neuronal cell.2. The neuronal cell according to claim 1 , wherein said polynucleotide is incorporated into said non-neuronal cell using a virus or viral vector.3. The neuronal cell according to claim 2 , wherein said virus or vector is adenovirus claim 2 , AAV claim 2 , retrovirus claim 2 , or lentivirus.4. The neuronal cell according to claim 1 , wherein said cell is a human cell.5. The neuronal cell according to claim 1 , wherein said non-neuronal cell is a fibroblast cell.6. The neuronal cell according to claim 1 , wherein said neuronal cell expresses one or more of tubulin claim 1 , MAP2 claim 1 , synapsin claim 1 , or NeuN.7. A method for screening a drug or compound for activity in treating or inhibiting Alzheimer's ...

SCREENING METHOD FOR PAIN SUPPRESSOR AND PHARMACEUTICAL COMPOSITION FOR PREVENTION OR TREATMENT OF PAIN

The present invention provides a screening method for a pain suppressor, which method comprises using FLRT3 to select a substance capable of inhibiting FLRT3 expression or a substance capable of inhibiting FLRT3 transport to the spinal cord. In addition, the present invention provides a pharmaceutical composition for prevention or treatment of pain, which pharmaceutical composition comprises, as an active ingredient, a substance capable of inhibiting FLRT3 expression or a substance capable of inhibiting FLRT3 transport to the spinal cord. 1. A screening method for a pain suppressor substance , the method comprising:(a) contacting cells that express FLRT3 with a test substance;(b) measuring the level of FLRT3 expression in the cells that express FLRT3 after (a);(c) comparing the level of FLRT3 expression measured in (b) to a control population of FLRT3-expressing cells, which were not contacted with the test substance; and(d) selecting a test substance as a pain suppressor substance when said test substance reduces the level of FLRT3 expression in said cells that express FLRT3 after contact with said test substance, as compared to said control population of FLRT3-expressing cells, which were not contacted with the test substance.213-. (canceled)14. The method according to claim 1 , wherein the pain suppressor substance is a substance that suppresses peripheral neuropathy-associated pain.15. The method according to claim 1 , further comprising selecting a test substance as a pain repressor substance that inhibits FLRT3 transport to the spinal cord.16. The method according to claim 1 , further comprising selecting a test substance as a pain repressor substance that inhibits FLRT3 binding to Unc5B.17. The method according to claim 1 , wherein:the test substance is contacted with dorsal root ganglion cells, which express FLRT3, of the dorsal root ganglion tissue or spinal cord;the level of FLRT3 expression in the dorsal root ganglion tissue or spinal cord is measured; ...

In vitro assays for inhibition of microglial activation

The present invention provides cell-based assays, including high throughput cell-based assays, for identification of candidate therapeutic agent with no known agents with the ability to inhibit microglial activation in vivo in response to different ligands.

METHOD AND COMPOSITION FOR GENERATING BASAL FOREBRAIN CHOLINERGIC NEURONS (BFCNS)

The invention relates to methods and compositions for developing basal forebrain cholinergic neurons (BFCNs) from stem cells, and in particular, BFCNs having repaired electrophysiological defects relating to one or more mutations in PSEN2, and to the use of such BFCNs in cell-based therapies to treat Alzheimer's disease. 1. A method of generating basal forebrain cholinergic neurons (BFCNs) comprising:culturing pluripotent stem cells (PSCs) in a basal media comprising an inhibitor of transforming growth factor beta (TGF-β) signaling and an activator of sonic hedgehog (Shh) signaling to induce neuroectodermal differentiation, wherein the basal media lacks basic fibroblast growth factor (bFGF), TGF-β, lithium chloride (Li—Cl), GABA and pipecolic acid, thereby generating BFCNs.2. The method of claim 1 , wherein the inhibitor is a SMAD inhibitor.3. The method of claim 2 , wherein the inhibitor is SB431542 claim 2 , LDN193189 or combination thereof.4. The method of claim 1 , wherein the activator is an agonist of smoothened protein.5. The method of claim 4 , wherein the activator is smoothened agonist (SAG) claim 4 , purmorphamine or combination thereof.6. The method of claim 1 , wherein culturing is performed for a duration of about 4 to 9 days.7. The method of claim 2 , wherein the inhibitor is present in the culture media from about day 2 to day 8.8. The method of claim 1 , further comprising selecting for CD271+ cells.9. The method of claim 8 , wherein selecting is performed after about 9 to 12 days of culturing.10. The method of claim 9 , further comprising culturing the CD271+ cells in a neuronal basal medium to generate neuronal embryoid bodies (NEBs).11. The method of claim 10 , wherein the CD271+ cells are cultured for about 7 days thereby generating NEBs.12. The method of claim 11 , further comprising:a) harvesting the NEBs; andb) dissociating cells of the NEBs and re-plating the dissociated cells as a monolayer.13. The method of claim 12 , wherein the re-plated ...

METHODS FOR IDENTIFYING ARTHROPOD REPELLENTS AND ATTRACTANTS, AND COMPOUNDS AND COMPOSITIONS IDENTIFIED BY SUCH METHODS

Provided herein are screening methods for identifying compounds for use as an arthropod repellent based on the masking or inhibition of the detection of the skin odor by a cpA neuron. Provided herein are also screening methods for identifying compounds for use as an arthropod attractant based on activation of the cpA neuron. Further provided are one or more compounds identified using the screening methods described herein, and compositions containing such compounds. 1a) providing a candidate compound and a skin odor;b) providing a sample comprising a neuron, wherein the neuron expresses at least one of Gr1, Gr2, and Gr3 or any orthologs thereof;c) contacting the candidate compound with the sample;d) measuring the detection of the skin odor by the neuron;e) comparing the detection of the skin odor by the neuron after contact with the candidate compound to the detection of the skin odor by the neuron in the absence of the candidate compound; andf) identifying a compound that is a repellent for at least one arthropod species by determining whether or not the candidate compound masks or inhibits the detection of the skin odor by the neuron.. A method for identifying a compound that is a repellent for at least one arthropod species, comprising: This application is a continuation application of International Application PCT/US14/029201, with an international filing date of Mar. 14, 2014, which claims priority to and the benefit of U.S. Provisional Patent Application No. 61/799,734, filed Mar. 15, 2013, the disclosures of which are incorporated herein by reference in their entireties.The present disclosure relates generally to the field of arthropod repellents and attractants, and more specifically to methods of identifying such repellents based on affecting the detection of human skin odors in arthropods.Blood-feeding insects, such as mosquitoes, transmit deadly pathogens like malaria parasites, dengue viruses, and filarial worms to hundreds of millions of people every ...

METHOD OF IDENTIFYING AGENTS THAT AFFECT MATURATION, SURVIVAL AND MYELINATION

The present invention discloses a method of identifying agents that affect maturation and survival of oligodendrocytes or myelination of neuronal cells using ex-vivo differentiated embryonic stem cells. 130.-. (canceled)31. A method of making enriched oligodendrocyte precursor cells (OPCs) , comprising:(a) culturing human embryonic stem cells (hES) in a medium comprising retinoic acid and Epidermal Growth Factor (EGF), thereby generating neurospheres;(b) contacting the neurospheres with an adherent substrate comprising an extracellular matrix;(c) following step (b), dissociating the neurospheres;(d) culturing the dissociated neurospheres in a medium comprising EGF and basic Fibroblast Growth Factor (bFGF);(e) following step (d), culturing the dissociated neurospheres in a medium lacking growth factors, thereby generating ex vivo differentiated neural cells;(f) selecting the ex vivo differentiated neural cells to be O4-positive or CD140-positive;(g) expanding, freezing, and thawing the selected neural cells in a medium comprising EGF and bFGF;(h) after thawing, culturing the selected neural cells for at least one day in a medium lacking growth factors; thereby making the enriched OPCs.32. The method of claim 31 , wherein the OPCs further express Hes5.33. The method of claim 31 , wherein the hES in step (a) are aggregated.34. The method of claim 31 , wherein the medium of step (a) further comprises noggin.35. The method of claim 31 , wherein the medium of step (e) comprises noggin.36. The method of claim 31 , wherein the selecting of step (f) is performed by magnetic sorting (MACS) or fluorescence activated cell sorting (FACS).37. A method of quantifying an effect of an agent on myelination of OPCs or on maturation or survival of oligodendrocytes claim 31 , or a combination thereof claim 31 , comprising:{'claim-ref': {'@idref': 'CLM-00031', 'claim 31'}, '(a) making enriched OPCs according to ;'}(b) following step (a), contacting the enriched OPCs with the agent; and,( ...

Methods and devices for live cell imaging analysis

Provided herein are methods for analysis of target cells on a population or individual basis, including before and after contact with a stimulus in order to determine the effect of such stimulus on the target cells. Also provided are devices for performing such methods. The analysis methods involve identifying and measuring or tracking morphological changes that occur in target cells over a period of time. Tracking is accomplished using imaging systems capable of imaging target cells individually over a period of time either continuously or at discrete intervals of time.

Determination of parkinson's disease

Methods and compositions for accurate identification of Parkinson's disease are disclosed. More particularly, the disclosure is directed to the determination of Parkinson's disease in ante-mortem tissue samples.

TREATMENT OF NEURODEGENERATIVE AND NEURODEVELOPMENTAL DISEASES BY INHIBITION OF THE a2-Na/K ATPase/a-ADDUCIN COMPLEX

Described herein are methods for the prevention of neurodegeneration and the treatment of neurodegenerative disease (Including amyotrophic lateral sclerosis) ami neurodevelopmental disorders through the administration of an agent that inhibits die a2-Na/K ATPase/a-Adducin Complex. 1. A method of treating a neurodegenerative disease or a neurodevelopmental disorder comprising administering to a subject an agent that inhibits α2-Na/K ATPase and/or α-Adducin.2. The method of claim 1 , wherein the disease is a neurodegenerative disease selected from the group consisting of amyotrophic lateral sclerosis (ALS) claim 1 , Huntington's disease claim 1 , spinocerebellar ataxias claim 1 , Alzheimer's disease claim 1 , traumatic brain injury and Parkinson's disease or a neurodevelopmental disorder selected from the group consisting of fragile X syndrome claim 1 , Down's syndrome claim 1 , Rett syndrome claim 1 , intellectual disability claim 1 , autism claim 1 , an autism spectrum disorder and Asperger syndrome.3. The method of claim 2 , wherein the neurodegenerative disease is ALS.4. The method of claim 1 , wherein the agent is selected from the group consisting of a small molecule claim 1 , an interfering nucleic acid molecule specific for α2-Na/K ATPase claim 1 , an antibody that binds to α2-Na/K ATPase claim 1 , an isolated soluble polypeptide comprising at least 5 consecutive amino acids of the amino acid sequence encoding α-Adducin claim 1 , an interfering nucleic acid molecule specific for α-Adducin claim 1 , and an isolated soluble polypeptide comprising at least 5 consecutive amino acids of the amino acid sequence encoding α2-Na/K ATPase.5. The method of wherein the small molecule is a cardiac glycoside.6. The method of claim 5 , wherein the small molecule is selected from the group consisting of digoxin claim 5 , ouabain claim 5 , digitoxin claim 5 , proscillaridin A claim 5 , digoxigenin claim 5 , gitoxin claim 5 , gitoxigenin claim 5 , oleandrin claim 5 , butalin ...

THERAPEUTIC CURCUMIN DERIVATIVES

Curcumin analogues and methods are provided for treatment of disease. 114-. (canceled)15. A method for identifying a therapeutic curcumin derivative , comprising:contacting a cell comprising at least one of NF-κB, AP-1 or GSTP1-1 with a curcumin derivative;contacting the cell with an activator of NF-κB, AP-1 or GSTP1-1; anddetermining the effect of the curcumin derivative on cell activation;wherein a curcumin derivative that reduces cell activation is identified as an therapeutic curcumin derivative.16. The method of claim 15 , wherein the activator comprises TNF-α claim 15 , IL-1 claim 15 , PMA or an MAPK kinase.17. The method of claim 15 , wherein the cell is an adipocyte or endothelial cell.18. The method of claim 15 , wherein the cell is a brain cell.19. The method of claim 18 , wherein the brain cell is a glial cell.20. A method for identifying a therapeutic curcumin derivative claim 18 , comprising:contacting a solution comprising an Aβ peptide with a curcumin derivative; anddetermining the effect of the curcumin derivative on aggregation of the Aβ peptide;wherein a curcumin derivative that reduces aggregation of the Aβ peptide is identified as a therapeutic curcumin derivative.21. The method of claim 20 , wherein the effect of the curcumin derivative on aggregation of the Aβ peptide is determined by an immunological assay.22. A method of treating a subject afflicted with cancer or a precancerous condition claim 20 , the method comprising administering to the subject a therapeutically effective amount of a composition comprising a curcumin derivative selected from the group consisting of:{'sup': 1', '2', '1', '2, '(a) Ar-L-Ar(Formula I); wherein L is a divalent linking group comprising an alkylene or an alkenylene comprising 3, 4, 5, 6, or 7 backbone carbon atoms, wherein one or more of the backbone carbon atoms form part of a carbonyl or secondary alcohol; and Arand Arare each independently aryl groups; and'}{'sup': 1', '11', '1', '11, '(b) Ar-L-R(Formula IV ...

METHOD FOR SCREENING THERAPEUTIC AND/OR PROPHYLACTIC AGENTS FOR ALZHEIMER'S DISEASE

The present invention provides a method for screening a therapeutic and/or prophylactic agent for Alzheimer's disease using at least one index selected from the levels of Aβ oligomers, BiP, cleaved caspase 4, PRDX4 and ROS in nerve cells or the like whose differentiation has been induced from iPS cells derived from somatic cells of a patient with Alzheimer's disease. 1. A method for screening a therapeutic agent and/or prophylactic agent for Alzheimer's disease , said method comprising the steps of:(a) bringing a candidate substance into contact with a nerve cell(s) or astrocyte(s) derived from an induced pluripotent stem (iPS) cell(s) prepared from a somatic cell(s) of a patient with Alzheimer's disease, or derived from an iPS cell(s) in which amyloid precursor protein (APP) having deletion mutation of glutamic acid at position 693 has been introduced;(b) measuring the amount of Aβ oligomers in said nerve cell(s) or astrocyte(s); and(c) selecting said candidate substance as a therapeutic and/or prophylactic agent for Alzheimer's disease if the amount of Aβ oligomers is decreased as compared to a case where said candidate substance is not brought into contact.2. The method according to claim 1 , wherein said nerve cell or astrocyte is a cell that accumulates Aβ oligomers.3. The method according to claim 1 , wherein said somatic cell of a patient with Alzheimer's disease is a somatic cell having APP having deletion mutation of glutamic acid at position 693.4. The method according to claim 1 , said method further comprising the steps of:(a) measuring at least one index selected from the group consisting of the ER stress level, caspase 4 activity, transgelin level and oxidative stress level in said nerve cell(s) or astrocyte(s); and(b) selecting said candidate substance as a therapeutic and/or prophylactic agent for Alzheimer's disease if said level(s) and/or activity is/are decreased as compared to a case where said candidate substance is not brought into contact.5. ...

Expansion and Differentiation of Inner Ear Supporting Cells and Methods of Use Thereof

This disclosure relates to methods for expanding inner ear supporting cells (e.g., Lgr5+ inner ear supporting cells) and differentiating inner ear supporting cells (e.g., Lgr5+ inner ear supporting cells) to inner ear hair cells (e.g., atonal homolog 1 (Atoh1)+ inner ear hair cells) and the use of the inner hear supporting cells and hair cells, e.g., for identifying candidate therapeutic compounds for the treatment of hearing loss and balance loss. Additionally, the methods described herein can be used in the treatment of a subject having hearing loss and balance loss that would benefit from increased proliferation and differentiation of inner ear supporting cells (e.g., Lgr5+ inner ear supporting cells). 1. A method of producing an expanded population of inner ear supporting cells , the method comprising contacting a population of inner ear supporting cells with one or more agents selected from the group consisting of:(a) a retinoid receptor signaling activator;(b) a Wnt signaling activator set forth in Table A;(c) a bone morphogenetic protein (BMP) signaling inhibitor set forth in Table B;(d) a cyclin-dependent kinase (CDK) activator set forth in Table C;(e) an E box-dependent transcriptional activator set forth in Table D;(f) a Notch signaling activator set forth in Table E;(g) a histone deacetylase (HDAC) inhibitor set forth in Table F;(h) a protein degradation inhibitor set forth in Table G;(i) a PI3K-Akt signaling inhibitor set forth in Table H; and(j) a cAMP response element binding protein (CREB) activator set forth in Table I,wherein the one or more agents are present in amounts sufficient to produce an expanded population of inner ear supporting cells.2. The method of claim 1 , wherein the Notch signaling activator is a Delta-like protein activator claim 1 , a Jagged protein activator claim 1 , a Notch activator claim 1 , and/or a γ-secretase activator.3. The method of claim 1 , wherein the one or more agents is selected from the group consisting of:(a) a ...

SCREENING METHOD FOR THERAPEUTIC AGENTS FOR CHARCOT-MARIE-TOOTH DISEASE AND SELF-DIFFERENTIATION MOTOR NEURONS USED THEREFOR

The present invention relates to a method for the screening of a therapeutic agent for Charcot-Marie-Tooth disease (CMT) using induced pluripotent stem cells and motor neurons differentiated therefrom. Particularly, the present inventors prepared induced pluripotent stem cells from the human fibroblasts originated from CMT patient. When the motor neurons differentiated from the said induced pluripotent stem cells are used for the screening of a therapeutic agent for Charcot-Marie-Tooth disease, the pharmaceutical effect of the therapeutic agent candidates can be easily evaluated during the screening. In addition, by the method to prepare the induced pluripotent stem cells, autologous motor neurons which are usable for the screening of a patient-specific therapeutic agent and the patient-specific treatment can be prepared. 1. A method for preparation of motor neurons from somatic cells originated from a Charcot-Marie-Tooth disease (CMT) patient , wherein the method comprises the following steps:1) obtaining human somatic cells from the Charcot-Marie-Tooth disease (CMT) patient;2) transfecting the human somatic cells originated from the CMT patient of step 1) with a vector comprising OCT4, SOX2, KLF4, and c-MYC transgenes, followed by culturing to induce induced pluripotent stem cells (iPSC); and3) culturing the induced pluripotent stem cells prepared in step 2) in the presence of retinoic acid and sonic hedgehog to induce motor neurons.2. A method for preparation of motor neurons from somatic cells originated from a Charcot-Marie-Tooth disease (CMT) patient , wherein the method comprises the following steps:1) obtaining human somatic cells from the Charcot-Marie-Tooth disease (CMT) patient;2) transfecting the human somatic cells originated from the CMT patient of step 1) with a vector comprising OCT4, SOX2, KLF4, and c-MYC transgenes, followed by culturing to induce induced pluripotent stem cells (iPSC);3) culturing the induced pluripotent stem cells prepared in step ...

DERIVATION OF HUMAN MICROGLIA FROM PLURIPOTENT STEM CELLS

The present invention relates to methods for deriving human hematopoietic progenitors, primitive macrophages, and microglial cells from human pluripotent stem cells. In particular, provided herein are highly efficient and reproducible methods of obtaining human primitive macrophages and microglia from human pluripotent stem cells, where the primitive macrophages and microglia can be suitable for clinically relevant therapeutic applications. 1. A method of obtaining human hematopoietic precursor cells , comprising culturing human pluripotent stem cells under normoxic conditions for about 24 hours , wherein the pluripotent stem cells are cultured on a substrate that promotes cell adhesion and in a culture medium consisting essentially of L-ascorbic acid-2-phosphate magnesium , sodium selenium , transferrin , insulin , NaHCO , fibroblast growth factor 2 (FGF2) , transforming growth factor beta 1 (TGFβ1) , and a Rho kinase (ROCK) inhibitor , whereby the cultured pluripotent stem cells differentiate into hematopoietic precursor cells (HPCs).2. The method of claim 1 , wherein the substrate that promotes cell adhesion comprises Tenascin-C.3. The method of claim 2 , wherein the Tenascin-C is recombinant human Tenascin-C.4. The method of claim 1 , wherein the ROCK inhibitor is selected from the group consisting of Y-27632 and Blebbistatin.5. A method of obtaining human myeloid progenitors claim 1 , comprising culturing human HPCs obtained according to the method of for about 3 to about 5 days in a culture medium comprising FGF2 claim 1 , a vascular endothelium growth factor (VEGF) claim 1 , thrombopoietin (TPO) claim 1 , stem cell factor (SCF) claim 1 , interleukin-6 (TL-6) claim 1 , and interleukin-3 (IL-3) claim 1 , wherein the hematopoietic progenitor cells differentiate into myeloid progenitors.6. A method of obtaining human primitive macrophages claim 3 , comprising culturing human myeloid progenitors obtained according to the method of in the presence of a culture ...

METHODS OF USING GRAPHENE AND GRAPHENE-RELATED MATERIALS FOR MANIPULATION OF CELL MEMBRANE POTENTIAL

Methods for use of structures based on graphene-related materials are disclosed. These structures can be utilized for manipulating the cell transmembrane potential in various biomedical applications. 1. A method of eliciting changes in a functional state of a biological target , the method comprising:providing at least one biological target;positioning the at least one biological target and at least one biointerface adapted to generate free charge carriers in response to exposure to electromagnetic radiation, the at least one biointerface comprising one or more materials comprising at least one graphene sheet, graphene oxide, reduced graphene oxide, graphite, graphite oxide, or combinations thereof, the at least one biological target comprising live cells positioned to contact or to be in sufficient proximity to the at least one biointerface such that cell membrane potentials of the live cells are remotely and reversibly manipulated by the free charge carriers generated by exposure of the at least one biointerface to electromagnetic radiation; andexposing the at least one biointerface to electromagnetic radiation.2. The method of claim 1 , wherein the live cells comprise one or more intact cells.3. The method of claim 1 , wherein said exposing the at least one biointerface to electromagnetic radiation comprises exposing the at least one biointerface to visible spectrum electromagnetic radiation claim 1 , radio frequency spectrum electromagnetic radiation claim 1 , infrared electromagnetic radiation claim 1 , terahertz frequencies spectrum electromagnetic radiation claim 1 , microwave spectrum electromagnetic radiation claim 1 , or combinations thereof.4. The method of claim 1 , wherein said exposing the at least one biointerface to electromagnetic radiation comprises exposing the at least one biointerface to visible light.5. The method of claim 1 , further comprising using the at least one biointerface for production of cells for stem cell research and cell ...

METHOD FOR DIFFERENTIATION INTO RETINAL GANGLION CELLS FROM STEM CELLS

Provided are a method of preparing retinal ganglion cells by differentiation of stem into retinal ganglion cells, retinal ganglion cells differentiated by the method, a method of screening for a death inhibitor or a proliferation promoter of retinal ganglion cells using the retinal ganglion cells differentiated by the method, a kit of screening for the death inhibitor or the proliferation promoter of retinal ganglion cells including the retinal ganglion cells differentiated by the method, a pharmaceutical composition for treating glaucoma or optic neuropathy including the retinal ganglion cells, a method of treating glaucoma or optic neuropathy including the step of administering the retinal ganglion cells to a subject suspected of having glaucoma or optic neuropathy, and a method of preparing a mature retinal ganglion cell line. 1. A method of preparing mature retinal ganglion cells by differentiation of stem cells into mature retinal ganglion cells , comprising:(a) culturing retinal progenitor cells in a medium comprising an IGF1R (insulin-like growth factor-1 receptor) activator and a Wnt signaling pathway activator to differentiate them into immature retinal ganglion cells; and(b) culturing the immature retinal ganglion cells in a medium whose composition does not comprise the Wnt signaling pathway activator from that of the medium of step (a).2. The method of claim 1 , wherein the medium of step (a) comprises an IGF1R activator claim 1 , a BMP (bone morphogenetic protein) signaling pathway inhibitor claim 1 , an FGF (fibroblast growth factor) signaling pathway activator claim 1 , and a Wnt signaling pathway activator.3. The method of claim 2 , wherein step (b) comprises culturing the immature retinal ganglion cells in a medium whose composition does not comprise the BMP signaling pathway inhibitor claim 2 , FGF signaling pathway activator claim 2 , and Wnt signaling pathway activator claim 2 , and comprises an Shh (sonic hedgehog) signaling pathway activator ...

Induced pluripotent stem cell model of noonan syndrome and use thereof

The present invention relates to an induced pluripotent stem cell (iPSC) model of Noonan syndrome, a preparation method thereof, and uses to study of the pathogenesis of Noonan syndrome and a therapeutic agent screening method. Particularly, induced pluripotent stem cells from dermal fibroblasts of a Noonan syndrome-patient (NS-iPSCs) were generated, and differentiated into embryoid bodies (EBs), neural rosettes and neural cells. These iPSCs exhibited the normal morphology while showed reduced differentiation potency compare to control cell lines. NS-iPSCs were developed into embryoid bodies and neural rosettes by naturally and chemically directed differentiation. Interestingly, embryoid bodies and neural rosettes induced via chemically directed differentiation exhibited normal morphology and expressed ectoderm, neural rosettes and neural marker genes similar to normal cells. Thus, the cellular model can be useful in analytical research to understand pathogenesis of Noonan syndrome and establish screening method of the therapeutic agent.

SUPPORTED IN VITRO DEVELOPED TISSUE CULTURE AND CULTURING METHODS

An elongated or fiber-supported multicellular aggregation of multipotent cells, wherein multipotent cells are arranged in an oblong or longish arrangement with an aspect ratio of a prolate dimension to a perpendicular dimension of at least 2:1, or supported by a fibrous structure, and wherein the aggregate contains cells at different stages of differentiation, and the aggregate contains polar cells; methods of generating such aggregates; methods of developing the aggregates further into tissue organoids and kits for such methods. 115-. (canceled)16. A method of generating an elongated or fiber-supported multicellular aggregation of neural lineage with neuronal differentiated cells comprising the steps of:a) providing a plurality of pluripotent or non-human totipotent cells (i) that are located in an oblong or longish arrangement adhered to a support, said support has a length of 20 μm to 20 mm and a diameter of 1 μm to 60 μm, wherein said support is a biocompatible polymer that is not a biopolymer or wherein said support is a protein, or (ii) that are arranged on a fibrous structured support, and said support has a length of 20 μm to 20 mm and a diameter of 1 μm to 60 μm, wherein said fibrous structured support is a biocompatible polymer that is not a biopolymer or wherein said fibrous structured support is a protein; andb) letting said cells grow and differentiate in said arrangement, wherein said cells form intercellular bonds and adhere to each other;wherein said cells are stimulated to differentiate by a contacting the cells with a neuronal growth or differentiation factor.17. The method of claim 16 , wherein the arrangement has an aspect ratio of a prolate dimension to a perpendicular dimension of at least 2:1.18. The method of claim 16 , wherein said support is non-porous or has a porosity of less than 5% (v/v) of the supports volume.19. The method of claim 16 , wherein said support is a polymer microfilament and/or is biocompatible but not bioactive.20. The ...

HIGH THROUGHPUT OPTICAL ASSAY OF HUMAN MIXED CELL POPULATION SPHEROIDS

The present disclosure provides a method of performing a functional assay on human spheroids, e.g., three-dimensional human cell spheroids using, in one embodiment, a fluorometric imaging plate reader. 1. An optical method to detect the effect of one or more compounds on spheroids , comprising:contacting one or more spheroids comprising human cells of uniform diameter and one or more test compounds; andoptically detecting the amount or change in oscillations of one or more of the spheroids.2. The method of wherein the one or more spheroids are in wells of a multi-well plate.3. The method of wherein each well has one spheroid.4. The method of wherein the wells are further contacted with a fluorescent molecule useful to detect calcium claim 2 , and the amount or change in fluorescence over time is detected in one or more wells.5. The method of wherein the amount or change in fluorescence detects a quantity of peaks of fluorescence claim 4 , an amplitude of one or more of the peaks claim 4 , peak spacing between one or more of the peaks claim 4 , a width of one or more peaks claim 4 , or any combination thereof.6. The method of wherein the one or more spheroids comprise neurons.7. The method of wherein the one or more spheroids comprise neurons and astrocytes.8. The method of wherein the one or more spheroids comprise heart claim 1 , liver claim 1 , kidney claim 1 , pancreas claim 1 , lung claim 1 , endothelial or epithelial cells.9. The method of wherein the one or more spheroids comprise cancer cells or immortalized cells.10. The method of wherein the one or more spheroids comprise microglial cells or oligodendrocytes.11. The method of wherein the one or more spheroids comprise pericytes and endothelial cells.12. The method of wherein the one or more spheroids comprise endothelial cells claim 1 , microglial cells claim 1 , neurons claim 1 , oligodendrocytic cells claim 1 , or any combination thereof.13. The method of wherein the cells are progenitor cells.14. The ...

Three-Dimensional Human Neural Tissues for CRISPR-Mediated Perturbation of Disease Genes

Described herein are tissue cultures and methods for culturing stem-cell derived neurons in three-dimensional culture. Also provided are methods for screening therapeutic agents, developing cell line models, identifying genes associated with a neurological disease, and identifying genes associated with neuronal cell phenotypes. 1. A tractable neural tissue culture , comprising neuronal and/or glial cells derived from a stem cell and a three dimensional (3D) matrix comprising a biological supporting material configured to decrease the distance in expression space of the neural tissue culture as compared to a target tissue.2. The tractable neural tissue culture of claim 1 , wherein the expression space is over one or more genes claim 1 , 10 or more genes claim 1 , 20 or more genes claim 1 , 30 or more genes claim 1 , 40 or more genes claim 1 , 50 or more genes claim 1 , 100 or more genes claim 1 , 500 or more genes claim 1 , or 1000 or more genes.3. The tractable neural tissue culture of claim 1 , wherein the expression space is over a set of genes.4. The tractable neural tissue culture of claim 3 , wherein the set of genes defines a cell pathway.5. The tractable neural tissue culture of claim 1 , wherein the expression space is over an entire transcriptome.6. The tractable neural tissue culture of claim 1 , wherein the distance is measured by a Euclidean distance claim 1 , pearson coefficient claim 1 , spearman coefficient claim 1 , or combination thereof.7. The tractable neural tissue culture of claim 1 , wherein changes in the distance in expression space are determined from single cell sequencing data from the neural tissue culture claim 1 , target tissue claim 1 , or both.8. A tractable neural tissue culture claim 1 , comprising neuronal and/or glial cells derived from a stem cell and a 3D matrix comprising a biological supporting material configured to generate a statistically significant shift in gene expression distribution of the neural tissue culture as ...

TREATING LYSOSOMAL STORAGE DISEASE

Provided herein are compositions, kits and methods related to the treatment of one or more lysosomal storage diseases (LSDs) in a subject and methods of identifying agents that are useful for the treatment of one or more LSDs. The compositions, kits, and methods are based on the novel discovery that agents effective for treating LSD must have two characteristics: they must inhibit cell death and reduce suppression of cell division caused by toxic substances that accumulate in LSD. Exemplary agents include polypeptides (e.g., IGF-1, VEGF) and small molecules (e.g., chlorotrianisene, clofoctal, colforsin, and tulobuterol). 1. A method of treating a lysosomal storage disease (LSD) in a subject comprising administering to the subject an effective amount of a first agent that inhibits cell death and reduces suppression of cell division of primary central nervous system precursor cells in the presence of one or more toxic substances , wherein the toxic substances are known to accumulate in cells affected by a lysosomal storage disease.2. The method of claim 1 , wherein the first agent is selected from the group consisting of IGF-1 claim 1 , VEGF claim 1 , chlorotrianisene claim 1 , clofoctal claim 1 , and colforsin.3. The method of claim 1 , wherein the first agent is a small molecule.4. The method of claim 3 , wherein the small molecule is selected from the group consisting of chlorotrianisene claim 3 , clofoctal claim 3 , and colforsin.5. The method of claim 1 , wherein the LSD is a sphingolipid storage disorder.6. The method of claim 5 , wherein the sphingolipid storage disorder is selected from the group consisting of gloiboid-cell leukodystrophy (Krabbe Disease) claim 5 , metachromatic leukodystrophy claim 5 , and Gaucher's Disease.7. The method of claim 1 , wherein the one or more toxic substances are selected from the group consisting of glucopsychosine claim 1 , lysosulfatide claim 1 , and psychosine.8. The method of claim 1 , wherein the first agent inhibits cell ...

PRODUCTION OF MIDBRAIN DOPAMINERGIC NEURONS AND METHODS FOR THE USE THEREOF

Methods are provided for efficient production of midbrain dopaminergic (DA) neurons. In some aspects, methods involve differentiation and selection of DA neurons for a transgenic pluripotent cell population (e.g., cells comprising a selectable marker gene). Cell populations produced by the instant methods and methods of their use are likewise provided. 144-. (canceled)45. A method for providing an enriched population of midbrain dopaminergic (DA) neurons comprising:differentiating cells of a population of human induced pluripotent cells to provide a neural lineage cell population;further differentiating cells of the neural lineage cell population to generate a cell population which includes midbrain neurons; andpurifying cells from said cell population using a transgenic screenable or selectable marker under the control of a pan-neural promoter expressed by cells of the cell population, to provide an enriched population of midbrain DA neurons;wherein the differentiation differentiation steps are carried out in a chemically defined medium that is free of feeder cells and feeder cell extracts and at least 80% of cells in the enriched population are positive for both Lmx1 and FoxA2 expression.46. The method of claim 45 , wherein the enriched population of midbrain dopaminergic (DA) neurons comprise mammalian midbrain dopaminergic (DA) neuronal cells claim 45 , at least about 85% the neuronal cells being positive for both LIM homeobox transcription factor 1 (Lmx1) and forkhead box A2 (FoxA2) expression.47. The method of claim 46 , wherein the neuronal cells comprise at least 500 claim 46 ,000 cells.48. The method of claim 47 , wherein the neuronal cells comprise at least 1 million cells.49. The method of claim 46 , wherein at least about 80% of cells in the enriched population are positive for TH expression.50. The method of claim 45 , wherein the pan-neural promoter is a TuJ-1 claim 45 , Map-2 claim 45 , Dcx claim 45 , Synapsin claim 45 , enolase 2 claim 45 , glial ...

USE OF P97 AS AN ENZYME DELIVERY SYSTEM FOR THE DELIVERY OF THERAPEUTIC LYSOSOMAL ENZYMES

The present invention provides for compositions and methods for treating, ameliorating or preventing a lysosomal storage disease by administering to a patient suffering front a lysosomal storage disease a P97 conjugated with art enzyme which is capable of transportation into the lysosomes of cells on either sides of the blood brain barrier. 1. A method for treating a subject having a lysosomal storage disease , said method comprisingadministering a pharmaceutical composition to the subject wherein the composition comprises a p97 molecule covalently linked to a protein whose deficiency causes the disease.213-. (canceled)14. A compound comprising a p97 molecule covalently linked to a protein whose deficiency causes a lysosomal storage disease.1520-. (canceled)21. A method of screening a compound for therapeutic activity in treating a lysosomal storage disease , said method comprising:contacting a cell having a lysosome with the compound, wherein the compound comprises p97 covalently linked to a protein deficient in a lysosomal storage disease; andmonitoring delivery of the compound to the lysosome.2229-. (canceled) This application claims priority benefit of U.S. Provisional Application No. 60/347,758 filed on Jan. 11, 2002 which is incorporated herein by reference in its entirety.The present invention is related to the field of pharmaceutics and specifically to treatment of diseases through the introduction of p97 protein or polypeptide conjugated to a therapeutic or diagnostic agent to a subject. In particular, this invention relates to conjugates of p97 and proteins deficient in a lysosomal storage disease and methods for the treatment, amelioration or prevention of lysosomal storage diseases by administration of the conjugates to subjects having such diseases.Lysosomal storage diseases (LSDs) result from the absence or reduced activity of specific enzymes within the lysosomes of a cell. A large number of these enzymes have been identified and correlated with their ...

REPROGRAMMING-BASED MODELS OF NEURODEVELOPMENTAL DISORDERS AND USES THEREOF

The present invention relates to iPSC produced from fibroblast obtained from a subject affected by a neurodevelopmental disorder entailing intellectual disability (ID) and/or a disorder belonging to the Autism Spectrum Disorder (ASD) and/or Schizophrenia (SZ) and uses thereof. The present invention also relates to a cortical neural progenitor cell or a terminally differentiated cortical glutamatergic or gabaergic neuronal cell or a neural crest stem cell line, a mesenchymal stem cell line produced from the iPSC or iPSC line. The invention also relates to method for identifying a compound for the treatment and/or prevention of a neurodevelopmental disorder entailing intellectual disability (ID) and/or a disorder belonging to the Autism Spectrum Disorder (ASD) and/or Schizophrenia (SZ) and to a LSD1 inhibitor or a HDAC2 inhibitor for use in the treatment of such disorders. 1. A population of induced pluripotent stem cells (iPSC) produced from a fibroblast obtained from a subject , wherein the subject is affected by at least one of: a neurodevelopmental disorder entailing intellectual disability (ID) , a disorder belonging to the Autism Spectrum Disorder (ASD) , and Schizophrenia (SZ); and wherein said iPSC expresses detectable levels of at least one of the following markers: OCT4 , SOX2 , NANOG , NGN2 , and KLF4.2. The iPSC population of claim 1 , wherein the subject is affected by at least one of: a disorder caused by symmetrical copy number variations (CNV) at 7q11.23; Williams Beuren syndrome (WBS claim 1 , OMIM 194050); 7q11.23 microduplication syndrome; Somerville-van der Aa syndrome (OMIM 609757); and autistic spectrum disorder (7dupASD).3. The iPSC population of claim 1 , wherein iPSCs within the iPSC population harbor at least one copy-number variation claim 1 , selected from the group consisting of: chr6:254253-381137 (−) claim 1 , chr20:29989418-30665270 (+) claim 1 , chr6:254175-381137 (−) claim 1 , chr6:254253-381137 (−) claim 1 , chr6:102252826-102423940 ...

INDOLE AND INDOLINE DERIVATIVES AND METHODS OF USE THEREOF

The present application relates to indole and indoline derivatives of formula (I), (II), (III), (IV), (V), or (VI) 127.-. (canceled)28. A compound , or a pharmaceutically acceptable salt thereof , selected from the group consisting of:2,11-dimethyl-5-[2-(6-methylpyridin-3-yl)ethyl]-5,6,7,8,9,10-hexahydro-7,10-epiminocyclohepta[b]indole;5-[6-(4-iodophenyl)pyridazin-3-yl]-2,11-dimethyl-5,6,7,8,9,10-hexahydro-7,10-epiminocyclohepta[b]indole;9-methyl-6-[2-(6-methylpyridin-3-yl)ethyl]-3,4,5,6-tetrahydro-1H-2,5-methanoazepino[4,3-b]indole;2,8-dimethyl-5-[2-(6-methylpyridin-3-yl)ethyl]-2,3,4,5-tetrahydro-1H-1,4-methanopyrido[4,3-b]indole;6,10-dimethyl-9-[2-(6-methylpyridin-3-yl)ethyl]-2,3,4,9-tetrahydro-1H-4,1-(epiminomethano)carbazole;2,6-dimethyl-9-[2-(6-methylpyridin-3-yl)ethyl]-2,3,4,9-tetrahydro-1H-1,4-methano-3-carboline;6,11-dimethyl-9-[2-(6-methylpyridin-3-yl)ethyl]-2,3,4,9-tetrahydro-1H-1,4-(epiminomethano)carbazole;2,11-dimethyl-5-[2-(6-methylpyridin-3-yl)ethyl]-5,6,7,8,9,10-hexahydro-6,9-epiminocyclohepta[b]indole;2,12-dimethyl-5-[2-(6-methylpyridin-3-yl)ethyl]-6,7,8,9,10,11-hexahydro-5H-6,10-epiminocycloocta[b]indole;6,10-dimethyl-9-[2-(6-methylpyridin-3-yl)ethyl]-2,3,4,9-tetrahydro-1H-1,4-epiminocarbazole;2,9-dimethyl-6-[2-(6-methylpyridin-3-yl)ethyl]-1,2,3,4,5,6-hexahydro-1,5-methanoazepino[4,3-b]indole;2,9-dimethyl-6-[2-(6-methylpyridin-3-yl)ethyl]-1,2,3,4,5,6-hexahydro-1,4-methanoazepino[4,3-b]indole;2,11-dimethyl-5-[2-(6-methylpyridin-3-yl)ethyl]-5,6,7,8,9,10-hexahydro-6,10-epiminocyclohepta[b]indole;(5aS*,7S*,10R*,10aR*)-2,11-dimethyl-5-[2-(6-methylpyridin-3-yl)ethyl]-5,5a,6,7,8,9,10,10a-octahydro-7,10-epiminocyclohepta[b]indole;(5aR*,7S*,10R*,10aS*)-2,11-dimethyl-5-[2-(6-methylpyridin-3-yl)ethyl]-5,5a,6,7,8,9,10,10a-octahydro-7,10-epiminocyclohepta[b]indole;(5aS*,7S*,11R*,11aR*)-2,12-dimethyl-5-[2-(6-methylpyridin-3-yl)ethyl]-5a,6,7,8,9,10,11,11a-octahydro-5H-7,11-epiminocycloocta[b]indole;(5aR*,7S*,11R*,11aS*)-2,12-dimethyl-5-[2-(6-methylpyridin-3-yl) ...

DIAGNOSTIC METHODS FOR NEURAL DISORDERS

The invention generally relates to optical methods for the diagnosis of neuronal condition by converting a cell from a patient into a neuron and optically evaluating action potentials of that cell in vitro. The cell is transformed with an optical reporter and exhibits an optical signature in response to neural stimulation. Using genome-editing, a control cell can be made that is isogenic but-for a known mutation and a control signature obtained from the control cell. Thus, methods of the invention reveal potential neurodegenerative effects of a mutation as manifested in a patient's genetic context. The optical signature of the cell, or the difference between the signature and the control signature, is correlated to a diagnosis of the neurodegenerative disease 120-. (canceled)21. A method for determining an effect of a compound a neurological condition , the method comprising:presenting a compound to a sample comprising a plurality of neurons, wherein at least one of the plurality of neurons expresses an optical reporter of membrane electrical potential;receiving, via a microscopy system, an optical signal generated by the optical reporter in response to optical stimulation of a light gated ion channel in the sample following presentation of said compound; andidentifying the compound as a candidate for treatment of the neurological condition based on said optical signal.22. The method of claim 21 , wherein a plurality of samples are exposed to a plurality of different compounds.23. The method of claim 21 , wherein the light gated ion channel comprises an algal channelrhodopsin being expressed by a second neuron in synaptic communication with the at least one of the plurality of neurons claim 21 , and the optical reporter of membrane potential comprises a microbial rhodopsin with between 1 and 10 amino acid substitutions relative to a wild type form of the microbial rhodopsin.24. The method of claim 23 , wherein the at least one of the plurality of neurons also ...

NON-INVASIVE, IN VITRO FUNCTIONAL TISSUE ASSAY SYSTEMS

Provided are functional cell and tissue assay systems based on substrate-integrated multifunctional microelectrode arrays implementing stem cell technology. The system covers normal and pathogenic characteristics. 133-. (canceled)34. An in vitro method for screening at least one test substance for an effect on an isolated population of cardiomyocyte cells , comprising:providing an electrode array comprising one or more isolated populations of cardiomyocyte cells having been obtained by differentiating mouse or human pluripotent stem cells, wherein the stem cells have been genetically altered to comprise a selectable marker operably linked to a regulatory sequence specific for a first cell type, wherein the one or more isolated population of cells have been differentiated and depleted of undifferentiated cells and/or of undesired cell types by using a selection system that is lethal to the undesired cells and cell types by expressing a selectable marker gene that renders cells of the first cell type resistant to the lethal effect,contacting the populations of cardiomyocyte cells in an electrode array with the at least one test substance; and (i) Na+ channel activity,', '(ii) Ca2+/K+ channel activity,', '(iii) K+ channel activity;', '(iv) amplitude and/or field potential duration,', '(v) chronotropy,', '(vi) arrhythmia,', '(vii) pH-value,', '(viii) oxygen partial pressure,', '(ix) beating arrest,', '(x) contractility,', '(xi) analysis of AV-dissociation contractility,', '(xii) conductivity and/or impedance,', '(xiii) nitrous oxide-effects, or', '(ix) morphological changes; and, 'measuring the electrical activity of the contacted cardiomyocyte cells with the electrode array and analyzing at least one parameter selected from the group consisting ofselecting a test substance that has an effect on at least one parameter in the measuring step as compared to cardiomyocyte cells of the same type which were not contacted with a test substance and wherein a change in said at ...

DEVICE

The present invention provides a nerve cell device in which early observation of nerve activity (spikes, bursts, and the like) is made possible and the measured electric strength is increased by cultivating neurons upon a cell scaffold. By using this nerve cell device, imaging of intracellular signaling is also possible. 1. A nerve cell device comprising a cell scaffold and neurons.2. The nerve cell device according to claim 1 , wherein the neurons are oriented.3. The nerve cell device according to claim 1 , wherein the cell scaffold is a fiber sheet formed of a polymeric material.4. The nerve cell device according to claim 3 , wherein the fiber sheet has an oriented structure claim 3 , a non-oriented structure or a mixed structure of orientation and non-orientation.5. The nerve cell device according to claim 3 , wherein the fiber sheet is coated with the extracellular matrix protein selected from polylysine claim 3 , polyornithine claim 3 , laminin claim 3 , fibronectin claim 3 , MATRIGEL® and GELTREX®.6. The nerve cell device according to claim 1 , wherein the neurons form a three-dimensional structure on the cell scaffold and/or in the cell scaffold.7. The nerve cell device according to claim 1 , wherein the neurons are neural cells derived from primary cultured cells or pluripotent stem cells.8. The nerve cell device according to claim 7 , wherein the neural cells derived from the primary cultured cells or pluripotent stem cells are neural cells derived from mammals.9. The nerve cell device according to claim 1 , wherein the neurons comprise glutamatergic claim 1 , dopaminergic claim 1 , gamma-aminobutyratergic claim 1 , monoaminergic claim 1 , histaminergic or cholinergic neurons.10. The nerve cell device according to claim 1 , wherein: the neurons are seeded at a density of 1×10cells/cmto 4×10cells/cmagainst the cell scaffold.11. The nerve cell device according to claim 1 , further comprising: a frame for holding the periphery of the nerve cell device.12. The ...

GENERATION OF PHOTORECEPTORS FROM HUMAN RETINAL PROGENITOR CELLS USING POLYCAPROLACTONE SUBSTRATES

The present invention relates to biocompatible compositions for transplantation into a sub-retinal space of the human eye. The compositions include a biodegradable polyester film, preferably a polycaprolactone (PCL) film, and a layer of human retinal progenitor cells. The compositions of the invention can be used as scaffolds for the treatment a number of ocular diseases, including retinitis pigmentosa and age-related macular degeneration. 1. A biocompatible composition comprising:a biodegradable and biocompatible polyester carrier film, anda layer of isolated human retinal progenitor cells and/or derivatives thereof adhered to at least a portion of the surface of said polyester film.2. The composition of wherein the polyester is selected from the group consisting of polylactic acid (PLA) claim 1 , polycaprolactone (PCL) claim 1 , polyesteramide (PEA) claim 1 , polyhydroxybutyrate (PHB) claim 1 , and derivatives and mixtures thereof.3. The composition of wherein the polyester is polycaprolactone (PCL).4. The composition of claim 1 , wherein the derivative of human progenitor cells comprise photoreceptor cells.5. The composition of claim 1 , wherein derivatives thereof comprise multipotent retinal cells.6. The composition of claim 1 , further comprising a coating material applied to the polyester film surface and located between the isolated cells and the polyester film surface.7. The composition of claim 6 , wherein the coating is a material selected from the group consisting of poly-D-lysine claim 6 , poly-L-lysine claim 6 , fibronectin claim 6 , laminin claim 6 , collagen I claim 6 , collagen IV claim 6 , vitronectin claim 6 , matrigel claim 6 , and mixtures thereof.8. The composition of claim 1 , wherein the isolated retinal progenitor cells are obtained from post-natal retinal tissue.9. The composition of claim 1 , wherein the isolated retinal progenitor cells are obtained from the fetal neural retina.10. The composition of claim 1 , wherein the polyester film ...

Optically sensitive cell network

A neural network is disclosed. The neural network comprises a plurality of optogenetically modified neural cells being three-dimensionally distributed in a hydrogel medium and being disconnected from any solid support having a shear modulus above 1 GPa.

METHOD OF INHIBITING MICROGLIAL CELL MIGRATION AND TREATING TRAUMATIC BRAIN INJURY

Disclosed herein is a method of inhibiting microglial cell migration in the brain of a mammalian subject in need thereof, including subjects suffering from the symptoms of traumatic brain injury (TBI) and subjects suffering from a hematoma in the brain comprising administering an effective amount streptolysin O (SLO) wherein microglial migration is inhibited providing therapeutic benefits to subjects suffering from diseases characterized by microglial cell migration and activation. 1. A method of inhibiting microglial cell migration in the brain of a mammalian subject in need thereof comprising administering an effective amount streptolysin O (SLO) wherein microglial migration is inhibited.2. The method of which inhibits microglial cell migration in the central nervous system of a mammalian subject suffering from a Tau pathology selected from the group consisting of primary age-related tauopathy claim 1 , chronic traumatic encephalopathy claim 1 , progressive supranuclear palsy claim 1 , corticobasal degeneration claim 1 , frontotemporal dementia claim 1 , parkinsonism linked to chromosome 17 claim 1 , Lytico-Botig disease claim 1 , ganglioglioma claim 1 , gangliocytoma claim 1 , meningioangiomatosis claim 1 , postencelphalitic parkinsonism claim 1 , subacute sclerosing panencephalatis claim 1 , lead encephalopathy claim 1 , tuberous sclerosis claim 1 , Hallervorden-Spatz disease and lipofuscinosis and Beta-amyloid diseases selected from the group consisting of Alzheimer's disease and prion mediated diseases claim 1 , CNS injuries selected from the group consisting of spinal cord injury and traumatic brain injury (TBI) claim 1 , neurodevelopmental diseases selected from the group consisting of schizophrenia claim 1 , stroke claim 1 , Amyotrophic Lateral Sclerosis claim 1 , cerebral ischemia and hematoma in the brain.3. The method of which inhibits microglial cell migration in a subject suffering from traumatic brain injury (TBI).4. The method of which treats one or ...

Composition and method for preventing or treating a tauopathy

The present invention is a composition and method for the prevention and treatment of a tauopathy. The composition of the invention includes N-terminal amino acid residues of the tau protein, which have been identified as being involved in toxic activation of a PP1/GSK3 signaling cascade and inhibition of fast axonal transport in human tauopathies.

Neuroprotective Chemicals and Methods for Identifying and Using Same

Provided herein are methods for identifying a compound having cell-protective (e.g., neuroprotective) activity. Compounds identified therefrom are also provided. These compounds can be used to treat various diseases, disorders, or conditions associated with, for example, unwanted cell death.

A novel nervous system-specific transmembrane proteasome complex that modulates neuronal signaling through extracellular signaling via brain activity peptides

The inventors surprisingly found that neural stimulation caused the synthesis and degradation of proteins into peptides which were then secreted into the cell media within minutes of stimulation by a novel neural-specific and membrane bound proteasome (neuronal membrane proteasome or NMP) that is transmembrane in nature. These secreted, activity-induced, proteasomal peptides (SNAPPs) range in size from about 500 Daltons to about 3000 Daltons. Surprisingly none of the peptides appear to be those previously known to have any neuronal function. Moreover, these SNAPPs have stimulatory activity and are heretofore a new class of signaling molecules. Moreover, the NMP appears to play a highly significant role in aspects of neuronal signaling known to be critical for neuronal function. The inventors have gone on to develop all tools to study this novel mechanisms including protocols and practice for generation and purification of SNAPPs as well as a new and specific inhibitor of the NMP allowing for selective control of this process in the nervous system. The present invention provides methods of making and using these SNAPPs for both laboratory and clinical purposes, the screening for molecules which modulate NMP function in vivo and in vitro, and methods for diagnosis of NMP related diseases.

MANUFACTURE OF STRUCTURE CAPABLE OF FORMING THREE-DIMENSIONAL NEURONAL SPHEROID AND GENERATING NEURITE THROUGH VARIOUS SURFACE PROCESSES

The present invention relates to the manufacture of a platform for forming a neuronal spheroid and, more specifically, to the manufacture of a structure capable of simultaneously forming a test-tube three-dimensional neuronal spheroid and generating a neurite by forming a three-dimensional neuronal spheroid with a micro-platform and forming a neurite between the neuronal spheroid and the micro-platform so as to enable signal transduction, which is an essential function of a nerve cell. 1. A platform for three-dimensional (3D) nerve cells culture , comprising concave microwells surface-treated with 3-aminopropyltriethoxysilane (APTES).2. The platform of claim 1 , wherein the concave microwells are hemispherical with a diameter of 100 μm to 1 claim 1 ,000 μm.3. The platform of claim 1 , wherein the platform for culturing 3D nerve cells promotes the formation of neuronal spheroids and neurites.4. The platform of claim 1 , wherein the substrate consists of a plurality of concave microwells claim 1 , wherein open spaces are present between the adjacent concave microwells.5. The platform of claim 1 , wherein the substrate is further surface-treated with one or more of carbon nanotubes (CNTs) claim 1 , laminin claim 1 , and poly-L-lysine (PLL).6. The platform of claim 1 , wherein the substrate consisting of the concave microwells is made of polydimethylsiloxane (PDMS).7. A method of culturing 3D nerve cells in vitro claim 1 , comprising:{'claim-ref': {'@idref': 'CLM-00001', '#text': 'claim 1'}, '#text': 'culturing nerve cells in the platform for culturing 3D nerve cells defined in .'}8. A method of screening a drug for treatment of damaged nerves or neurological disorders claim 1 , comprising:{'claim-ref': {'@idref': 'CLM-00001', '#text': 'claim 1'}, '#text': 'culturing nerve cells in the platform for culturing 3D nerve cells defined in ;'}treating the cultured nerve cells with a candidate substance;treating the nerve cells treated with the candidate substance with a ...

CELLULAR TEST SYSTEMS FOR THE DETERMINATION OF THE BIOLOGICAL ACTIVITIES OF NEUROTOXIN POLYPEPTIDES

The present invention pertains to a method for the generation of neurotoxin-sensitive, neuronal differentiated cells comprising the steps of: a) cultivating tumor cells which are able to differentiate into neuronal cells in a culture medium under conditions and for a time which primes said tumor cells for neuronal differentiation; and b) cultivating the tumor cells primed for neuronal differentiation of a) in a differentiation medium having an osmolality of 100 to 270 mOsm/kg, and comprising (i) B27 supplement and/or (ii) N2 supplement, for at least 3 days, thereby obtaining neurotoxin-sensitive, neuronal differentiated cells. The invention further relates to neurotoxin-sensitive, neuronal differentiated cells obtainable by the method of the invention. In addition, the invention encompasses a method for determining the activity of a neurotoxin polypeptide comprising the steps of: a) contacting the neurotoxin-sensitive, neuronal differentiated cells obtainable by the method of the invention with a neurotoxin polypeptide; b) cultivating the neurotoxin-sensitive, neuronal differentiated cells of step a) for 3 to 74 hours or 72 hours under conditions which allow for the neurotoxin polypeptide to exert its biological activity; and c) determining the activity of the neurotoxin polypeptide in the said cells after cultivation according to step b). Finally, the invention provides for a medium comprising OptiMEM, FBS, B27 supplement, and N2 supplement. 1. A method for determining the activity of a neurotoxin polypeptide comprising the steps of:a) cultivating SiMa or P19 tumor cells in a culture medium comprising 80 to 98.8% OptiMEM, 1 to 10% Fetal Bovine Serum (FBS), 0.2 to 5% B27 supplement and/or 0.2 to 5% N2 supplement and, optionally, non-essential amino acids and/or an antibiotic under conditions and for a period of time which primes SiMa or P19 tumor cells for neuronal differentiation;b) adjusting the osmolality of a differentiation medium comprising 78 to 98.3% ...

CHANNELRHODOPSINS FOR OPTICAL CONTROL OF CELLS

The invention, in some aspects relates to compositions and methods for altering cell activity and function and the introduction and use of light-activated ion channels. 1. A nucleic acid molecule , wherein the sequence of the nucleic acid molecule encodes a light-activated ion channel polypeptide comprising an amino acid sequence having at least 90% amino acid identity to amino acids 86-320 of SEQ ID NO: 2 and at least 95% identity to the remaining amino acids of SEQ ID NO: 2 , and wherein the nucleic acid molecule is in a vector.2. The nucleic acid molecule of claim 1 , wherein the sequence of the nucleic acid molecule encodes a light-activated ion channel polypeptide comprising an amino acid sequence having at least 95% identity to amino acids 86-320 of SEQ ID NO: 2 and at least 95% identity to the remaining amino acids in the sequence set forth as SEQ ID NO: 2.3. The nucleic acid molecule of claim 1 , wherein the sequence of the nucleic acid molecule encodes a light-activated ion channel polypeptide comprising an amino acid sequence having at least 99% identity to amino acids 86-320 of SEQ ID NO: 2 and 95% identity to the remaining amino acids in the sequence set forth as SEQ ID NO: 2.4. The nucleic acid molecule of claim 1 , wherein the sequence of the nucleic acid molecule encodes a light-activated ion channel polypeptide comprising an amino acid sequence having at least 99% amino acid identity to amino acids 86-320 of SEQ ID NO: 2 and at least 98% identity to the remaining amino acids of SEQ ID NO: 2.5. The nucleic acid molecule of claim 1 , wherein the sequence of the nucleic acid molecule encodes a light-activated ion channel polypeptide comprising an amino acid sequence having at least 99% amino acid identity to amino acids 86-320 of SEQ ID NO: 2 and at least 99% identity to the remaining amino acids of SEQ ID NO: 2.6. The nucleic acid molecule of claim 1 , wherein the sequence of the nucleic molecule encodes the amino acid sequence set forth as SEQ ID NO: ...

METHOD FOR IDENTIFYING EFFECTIVE TREATMENTS AGAINST NEURODEGENERATIVE DISORDERS

The present invention relates to systems and methods for reprogramming cells to generate neurons and identifying effective treatments for neurodegenerative disorders using such neurons, as well as systems and methods for treating and developing treatments for one or more neurodegenerative disorders. 1. A method for identifying a suitable treatment for a patient having a neurodegenerative disorder comprising:(i) generating neurons from (i) induced pluripotent stem cells from somatic cells or (ii) fibroblasts obtained from a patient;(ii) testing two or more prospective treatments against separate groups of the generated neurons; and(iii) assessing the neurons following each treatment.2. The method of claim 1 , wherein the neurons are generated by morphogen directed differentiation claim 1 , linear conversion claim 1 , or any combination of any of the foregoing.3. The method of claim 1 , wherein the fibroblasts are reprogrammed with at least one transcription factor and optionally a TGF-beta inhibitor claim 1 , a Ras mutant claim 1 , a p53 mutant lacking a DNA-binding domain claim 1 , or any combination of any of the foregoing.4. The method of claim 3 , wherein the TGF-beta inhibitor is RepSox claim 3 , the Ras mutant is hRasG12V claim 3 , and the p53 mutant is p53DD.5. The method of claim 3 , wherein the fibroblasts are reprogrammed with one or more of the transcription factors Lhx3 claim 3 , Ascl1 claim 3 , Brn2 claim 3 , Myt11 claim 3 , Isl1 claim 3 , Ngn2 and NeuroD1.6. The method of claim 1 , wherein the prospective treatments in step (ii) are selected from a library of known treatments for the neurodegenerative disorder.7. The method of claim 1 , wherein one or more of the prospective treatments in step (ii) include treatment by inhibition of PIKFYVE.8. The method of claim 1 , wherein one or more of the prospective treatments in step (ii) include administration of a PIKFYVE inhibitor.9. The method of claim 1 , wherein two or more of the prospective treatments in ...

IDENTIFICATION OF OLFACTORY RECEPTORS SENSITIVE TO DIFFERENT ODORANTS

This disclosure provides methods for identification of olfactory sensosry neurons (OSN) that recognize specific odorants. The method comprises introducing into the OSN, a permanent activity marker which exhibits a detectable and permanent change upon activation of the neuron in response to an odorant and exposure to an exteranal stimulus, such as light. The OSN can be isolated and its receptor characterized. 1. A method for identification of olfactory sensosry neurons (OSN) that recognize an odorant comprising the steps of:a) providing a population of OSNs, each OSN expressing an olfactory receptor (OR) and a permanent activity marker (PAM), wherein the PAM has has been introduced into the OSN and exhibits a permanent and detectable change upon activation of the OSN in response to an odorant and exposure to an external stimulus, wherein the external stimulus is light;b) exposing the population of OSNs to a test odorant and the external stimulus that will cause the PAM to exhibit the detectable change if the OSN containing the PAM is activated in response to the odorant;c) isolating the OSNs exhibiting the detectable change in PAM from the OSNs that do not exhibit the detectable change in PAM;d) identifying the OR or ORs expressed in the isolated OSNs.2. The method of claim 1 , wherein the PAM is a fluorescent protein which fluoresces or changes its fluorescence when exposed to light that comprises a wavelength of about 340 nm to about 420 nm.3. The method of claim 2 , wherein OSNs in step c) are isolated by fluorescence activated cell sorting.4. The method of claim 1 , wherein the PAM comprises:an EosFP polypeptide, which includes a C-terminus portion comprising the sequence of SEQ ID NO. 2 and an N-terminus portion comprising the sequence of SEQ ID NO. 3, or a variant and/or fragment thereof;a calmodulin (CaM) polypeptide, comprising the sequence of SEQ ID NO. 4, or a variant and/or fragment thereof; anda M13 polypeptide, comprising the sequence of SEQ ID NO. 5, or ...

Leptin Compositions and Methods for Treating Progressive Cognitive Function Disorders Resulting from Accumulation of Neurofibrillary Tangles and Amyloid Beta

The present disclosure provides compositions containing a leptin product and methods of clinical therapy and diagnostic methods for progressive cognitive disorders. According to one aspect, the described invention provides a method for treating a progressive cognitive disorder. According to another aspect, the described invention provides a method for improving resilience of cognitive function in a subject in need thereof. According to another aspect, the described invention provides a method for identifying an effective therapeutic agent for treating a progressive cognitive dysfunction disease or disorder that results from at least one of accumulation of Aβ, hyperphosphorylation of tau, or accumulation of neurofibrillary tangles.

CELLULAR DISCOVERY PLATFORM FOR NEURODEGENERATIVE DISEASES

In some aspects, a cross-species platform useful for drug discovery in neurodegenerative diseases is described. 1. A method of identifying a candidate therapeutic agent for treatment of a neurodegenerative disease comprising: (a) contacting a human neuron or glial cell that has a genotype associated with the neurodegenerative disease with an agent that reduces toxicity of a neurodegeneration associated protein associated with the disease in a yeast cell that expresses the neurodegeneration associated protein or that reduces the level of a phenotype associated with loss of function of a yeast homolog of a human loss-of-function neurodegenerative disease gene; (b) measuring the level of a phenotype associated with the disease in the neuron or glial cell; and (c) identifying the agent as a candidate therapeutic agent for treatment of the neurodegenerative disease if the level of the phenotype is reduced as compared with the level of the phenotype in the absence of the agent.2. The method of claim 1 , wherein the neurodegeneration associated protein comprises an alpha-synuclein protein claim 1 , TDP-43 protein claim 1 , FUS protein claim 1 , A-beta protein claim 1 , or polyglutamine-expanded protein.3. The method of claim 1 , wherein the neurodegenerative disease is a synucleinopathy claim 1 , TDP-43 proteinopathy claim 1 , FUS-opathy claim 1 , Alzheimer's disease claim 1 , or a polyglutamine expansion disease.4. The method of claim 1 , wherein the neuron or glial cell is not engineered to have an extra copy of a gene encoding the neurodegeneration associated protein and has not been derived or cultured in the presence of an agent that enhances the phenotype.5. The method of claim 1 , wherein the phenotype is detectable in the yeast cell in the absence of the agent.6. The method of claim 1 , wherein the phenotype is detectable in a neuron or glial cell derived from an iPS cell derived from a human subject who has the disease claim 1 , wherein the neuron or glial cell is ...

DEVICES AND METHODS COMPRISING NEUROMUSCULAR JUNCTIONS

Disclosed herein are devices comprising one or more cantilevers comprising one or more neuromuscular junctions formed by a co-culture of myotubes and motoneurons. Disclosed herein are methods of using the disclosed devices comprising one or more cantilevers. This abstract is intended as a scanning tool for purposes of searching in the particular art and is not intended to be limiting of the present invention. 1. A device , comprising:at least one cantilever, wherein the at least one cantilever comprises one or more neuromuscular junctions formed by a co-culture of myotubes and motoneurons; andan automated detection system configured to detect a deflection of the at least one cantilever.2. (canceled)3. (canceled)4. The device of claim 1 , wherein the automated detection system comprises a laser and a photo-detector.5. The device of claim 1 , wherein muscle cells or myoblasts fuse together to form one or more of the myotubes.6. The device of claim 1 , wherein the at least one cantilever comprises one or more piezoelectric materials.7. The device of claim 6 , wherein the automated detection system comprises a transducer for detecting a change in electrical conductivity of the at least one cantilever.8. The device of claim 6 , wherein piezoelectric materials comprise at least one of quartz claim 6 , bone claim 6 , sodium tungstate claim 6 , zinc oxide claim 6 , lead zirconate titanate claim 6 , or a combination thereof.9. The device of claim 1 , wherein the at least one cantilever comprises at least two cantilevers.10. The device of claim 4 , wherein the automated detection system further comprises a plurality of linear actuators attached to XY translational stages that control the position of the laser and the photo-detector.11. The device of claim 4 , further comprising a digitizer and a computer claim 4 , wherein the photo-detector is in communication with the digitizer and wherein the digitizer is in communication with the computer.12. The device of claim 1 , ...

SLEEP MODULATION AGENT

The present invention relates to one or more ligands of a potassium channel β subunit for use in therapy, and in particular for use in treating or preventing a sleep disorder in a subject. The invention also provides a method of screening a test compound to determine if it is a substrate of a potassium channel β subunit. 1. A ligand of a potassium channel β subunit for use in therapy.2. A ligand of a potassium channel β subunit for use in treating or preventing a sleep disorder in a subject.3. The ligand for use according to or , wherein the ligand is a ligand of an aldo-keto-reductase domain of the β subunit of a potassium channel.4. The ligand for use according to any preceding claim , wherein the ligand is a ligand of a (3 subunit of a potassium channel comprising a Kv1 , Kv2 , Kv3 , Kv4 , Kv5 , Kv6 , Kv7 , Kv8 , Kv9 , Kv10 , Kv11 or Kv12 α subunit.5. The ligand for use according to any preceding claim , wherein the ligand is a ligand of a β subunit of a potassium channel comprising a Kv1.1 , Kv1.2 , Kv1.3 , Kv1.4 , Kv1.5 , Kv1.6 , Kv1.7 or Kv1.8 α subunit.6. The ligand for use according to any preceding claim , wherein the ligand is a ligand of a Kvβ1 , Kvβ2 or Kvβ3 subunit.7. The ligand for use according to any preceding claim , wherein the ligand does not bind to the aldo-keto-reductase domain.8. The ligand for use according to any preceding claim , wherein the ligand binds to the active site of the aldo-keto-reductase domain.9. The ligand for use according to any preceding claim , wherein the ligand is an allosteric ligand.10. The ligand for use according to claim 3 , wherein the ligand is a substrate of the aldo-keto-reductase.11. The ligand for use according to claim 10 , wherein the substrate is an electron acceptor.12. The ligand for use according to or claim 10 , wherein the substrate contains a carbonyl functional group (e.g. an aldehyde or a ketone).13. The ligand for use according to any one of to claim 10 , wherein the substrate is 4-oxo-2-nonenal (4 ...

TRANSGENIC ANIMAL MODEL OF MOOD DISORDERS

A non-human transgenic animal having a polynucleotide encoding a PTN polypeptide, which polynucleotide is operably linked to a promoter, wherein said transgenic animal has greater than wild-type expression of the PTN polypeptide in at least one brain region, as well as related vectors, methods of producing transgenic animals, in vitro and in vivo screening methods for potential therapeutic agents, and methods for treating and diagnosing neuropsychiatric illnesses, particularly anxiety and depression, are disclosed. 1. A non-human transgenic animal having a polynucleotide encoding a PTN polypeptide , which polynucleotide is operably linked to a neuron-specific promoter , wherein said transgenic animal has greater than wild-type expression of the PTN polypeptide in at least one brain region , and wherein said neuron-specific promoter is selected from the group consisting of: a Thy1 gene promoter , Neuron specific enolase (NSE) gene promoter; Rhombotin I gene promoter; PGK gene promoter; Neurofilament Low (NF-L) gene promoter; dopamine beta-hydroxylase (DBH) gene promoter; and Synapsin-1 gene promoter.2. A transgenic animal according to claim 1 , wherein said polynucleotide encodes:(i) a PTN polypeptide having an amino acid sequence having at least 80% amino acid sequence identity to the sequence of SEQ ID NO: 2;(ii) a PTN polypeptide having the amino acid sequence of SEQ ID NO: 2(iii) a PTN polypeptide having an amino acid sequence having at least 80% amino acid sequence identity to the sequence of SEQ ID NO: 4;(iv) a PTN polypeptide having the amino acid sequence of SEQ ID NO: 4; or(v) an active fragment of any one of (i)-(iv) having at least 100 amino acids, and wherein said PTN polypeptide of any one of (i)-(iv) or said active fragment of (v) is capable of binding to specific receptors RPTPβ/ζ, ALK or Syndecan 3 polypeptide.3. A transgenic animal according to or claim 1 , wherein the neuron-specific promoter is a Thy1 gene promoter.4. A transgenic animal according ...

NOCICEPTOR-LIKE CELLS DIFFERENTIATED FROM HUMAN NEURAL PROGENITORS AND USES THEREOF

Disclosed herein are methods of differentiating human neural progenitor cells to nociceptor-like cells. Also disclosed are methods of making an innervated skin-like construct using nociceptor-like cells differentiated from human neural progenitor cells. Also disclosed are engineered constructs for screening potentially therapeutic compounds that include a skin-like construct and nociceptor-like cells differentiated from human neural progenitor cells. Also disclosed is a method of screening potential therapies. 1. A method of differentiating human neural progenitor cells to nociceptor-like cells , comprising:a) exposing human neural progenitor cells to a first serum-free initiation medium for a first time period; andb) exposing the human neural progenitor cells to a second serum-free initiation medium for a second time period; andc) exposing the human neural progenitor cells to a serum-free differentiation medium during a third time period to cause at least a portion of the human neural progenitor cells to differentiate into nociceptor-like cells;wherein the nociceptor-like cells possess at least one nociceptor-like property.2. The method of claim 1 , wherein the first serum-free initiation medium comprises KSR base medium claim 1 , SB43152 claim 1 , and LDN-193189.3. The method of claim 2 , wherein the concentration of SB43152 in the first serum-free initiation medium is from 1-100 μM claim 2 , and wherein the concentration of LDN-193189 in the first-serum free initiation medium is from 10-1000 nM.4. The method of claim 3 , wherein the SB43152 concentration in the first-serum free initiation medium is about 10 μM claim 3 , and wherein the concentration of LDN-193189 in the first-serum free initiation medium is about 100 nM.5. The method of claim 1 , wherein the first time period is from 1-5 days.6. The method of claim 5 , wherein the first time period is about 2 days.7. The method of claim 1 , wherein the second serum-free initiation medium comprises SB43152 claim 1 ...

Method and System for Imaging and Analysis of a Biological Specimen

The present disclosure provides methods of preparing a biological specimen for imaging analysis, comprising fixing and clearing the biological specimen and subsequently analyzing the cleared biological specimen using microscopy. Also included are methods of quantifying cells, for example, active populations of cells in response to a stimulant. The present disclosure also provides devices for practicing the described methods. A flow-assisted clearing device provides rapid clearing of hydrogel-embedded biological specimens without the need of specialized equipment such as electrophoresis or perfusion devices.

METHODS FOR THE IDENTIFICATION, ASSESSMENT, PREVENTION, AND TREATMENT OF NEUROLOGICAL DISORDERS AND DISEASES USING FNDC5

The invention provides methods for identifying, assessing, preventing, and treating neurological disorders and diseases using Fndc5 and modulators of Fndc5 expression or activity. 111-. (canceled)12. A method for assessing whether a subject is afflicted with a neurological disease or disorder or has a risk of developing a neurological disease or disorder comprising the steps of detecting the expression of the Fndc5 gene or the expression or activity of Fndc5 polypeptide in a sample of a subject , wherein a decrease in the expression of the Fndc5 gene or a decrease in the expression or activity of the Fndc5 polypeptide compared to a control indicates the presence of a neurological disease or disorder or the risk of developing a neurological disease or disorder in the subject , optionally wherein the sample is selected from the group consisting of whole blood , serum , plasma , saliva , cerebrospinal fluid , spinal fluid , and neural tissue.13. (canceled)14. The method of claim 12 , wherein the expression of the Fndc5 polypeptide is detecteda) using a reagent which specifically binds with the protein, optionally wherein the reagent is selected from the group consisting of an antibody, an antibody derivative, and an antibody fragment; orb) by detecting the presence in the sample of a transcribed polynucleotide or portion thereof, optionally i) wherein the transcribed polynucleotide is an mRNA or a cDNA, ii) wherein the step of detecting further comprises amplifying the transcribed polynucleotide, or iii) wherein the level of expression of the marker in the sample is assessed by detecting the presence in the sample of a transcribed polynucleotide which anneals with Fndc5 or anneals with a portion of an Fndc5 polynucleotide under stringent hybridization conditions.15. The method of claim 12 , wherein the Fndc5 polypeptide is selected from the group of polypeptides consisting of:a) a polypeptide encoded by a nucleic acid molecule comprising a nucleotide sequence encoding ...

PROCESS AND DEVICE FOR THE DETERMINATION OF ALTERATIONS IN NEURONAL CONNECTIVITY AND/OR MORPHOLOGY

The bioassay process for the determination of alterations in neuronal connectivity and/or morphology comprises the operations of:—identifying the critical distance corresponding to which a pair of spaced single neuronal cells, subjected to the assay, is still able to form reciprocally interconnecting neuritic extensions, after a predetermined incubation period under conditions which favour the formation of such neuritic extensions, wherein each of said single neuronal cells is adhered to a respective spot of a substance promoting the adhesion of said neuronal cells, said spots being deposited at a distance from one another onto a substrate inhibiting the adhesion of neuronal cells, and—comparing the critical distance identified with the critical distance determined, under the same conditions, for reference cells, wherein a decrease in said critical distance for the neuronal cells subjected to the assay, relative to the critical distance for the reference cells, is indicative of an alteration in the neuronal connectivity and/or morphology. 2. A process according to claim 1 , characterized in that it comprises the operations of:a) producing on a substrate inhibiting the adhesion of neuronal cells a plurality of pairs of spots of a substance promoting the adhesion of said neuronal cells, wherein each of said spots is separate and distinct from the other spots and wherein at least two of said pairs exhibit a different distance between the spots;b) plating and allowing to adhere correspondingly to each of said spots a single neuronal cell subjected to the assay;c) detecting, after said predetermined incubation period, the possible formation of neuritic extensions interconnecting adjacent neuronal cells, adherent to the spots of said pairs; andd) determining the maximum distance between the spots of said pairs corresponding to which the formation of the interconnecting neuritic extensions occurs, by comparison of the neuritic extension behaviour between pairs of spots at ...

INVOLUNTARY RESPONSE STIMULATOR & THERAPY

An apparatus for simulating hand tremors caused by Essential Tremor and Parkinson's disease which may also be used as part of a therapeutic program in association with surgically implanted stem cells introduced into a patient's substantia nigra. 1. A method of providing physical therapy session to a patient who has had undifferentiated or differentiated stem cells surgically introduced into said patient's substantia nigra , said method comprising:providing a therapeutic apparatus, said therapeutic apparatus comprising: a waveform generator operable to output a waveform; an electric driver connected to said waveform generator, wherein said electric driver is operable to receive as an input said waveform, and wherein said electric driver outputs a drive signal, wherein said drive signal varies in response to changes in said waveform; an actuator comprising a connection point, wherein said actuator is connected to said electric driver, wherein said actuator is operable to receive as an input said drive signal, and wherein said actuator is operable to displace said connection point along an axis in response to said drive signal; a cuff configured to encircle a portion of said patient's hand or arm, wherein operation of said actuator displaces said cuff along said axis; and a connecting mechanism attached to said connection point and to said cuff; a means for monitoring a dopamine level of said patient when said patient's hand or arm is placed inside of said cuff and said apparatus in in operation, wherein said monitoring means generates a monitoring means output; and a control system connected to said monitoring means, wherein said monitoring means output is an input to said control system, wherein said control system is connected to said waveform generator, and wherein said control system is operable to modify the amplitude and frequency of said waveform to maximize said dopamine level of said individual during use of said apparatus;securing a wrist or arm of said ...

Method and Medium for Amplifying Neural Precursor Cells

The present invention relates to a method for amplifying a population of neural precursors comprising the step of culturing neural precursors in the presence of a PKA inhibitor. 1. Method for amplifying a population of neural precursors comprising the step of culturing neural precursors in the presence of a PKA inhibitor.2. The method according to wherein said PKA inhibitor is a selective PKA inhibitor.3. The method according to wherein said PKA inhibitor is a non-selective PKA inhibitor.4. The method according to wherein said PKA inhibitor is selected from the group consisting of H7 claim 1 , H8 claim 1 , H9 claim 1 , H89 claim 1 , HA 1077 and HA 1004.5. The method according to wherein said PKA inhibitor is HA 1004 and is used at a concentration comprised between 20 μM and 40 μM.6. The method according to claim 1 , wherein the step of culturing neural precursors is carried out in a culture medium comprising:a source of carbon as energy substrate, such as glucose, galactose or sodium pyruvate;essential amino-acids;at least one vitamin;at least a purine and a pyrimidine as nucleic acid precursors;inorganic salts;at least one molecule known to limit natural ageing;at least one antioxidant and/or at least one enzyme involved in Reactive Oxygen Species detoxification;at least one phospholipid precursor;at least one unique fatty acid;at least one carrier protein; andoptionally, at least one additional protein or peptide.7. The method according to claim 1 , wherein said culture medium is N2B27 medium comprising DMEM/F12 and Neurobasal media in a 1/1 ratio claim 1 , N2 supplement (1/100) claim 1 , B27 supplement (1/50) and beta-mercaptoethanol (1/1000).8. The method according to claim 1 , wherein the neural precursors are human neural precursors.9. The method according to claim 1 , wherein the neural precursors are derived from embryonic stem cells (ES cells) or from induced pluripotent stem cells (iPS cells).10. A method for obtaining neural precursors comprising the ...

DIRECTED DIFFERENTIATION OF ASTROCYTES FROM HUMAN PLURIPOTENT STEM CELLS FOR USE IN DRUG SCREENING AND THE TREATMENT OF AMYOTROPHIC LATERAL SCLEROSIS (ALS)

The present invention discloses a method of identifying agents that affect human astrocytes functionality using ex-vivo differentiated pluripotent stem cells (PSC). In addition, the use of human progenitor astrocytes or human astrocytes for the treatment of Amyotrophic Lateral Sclerosis (ALS) in a human subject is also disclosed. 1. A method of screening an agent for preventing or treating Amyotrophic Lateral Sclerosis (ALS) the method comprising:(a) contacting a population of astrocytes, the astrocytes having been ex-vivo differentiated from pluripotent stem cells (PSC), with the agent;(b) co-culturing the population of astrocytes of step (a) or a conditioned medium thereof with a population of neurons; and(c) quantifying an effect of said agent to enhance survival or neural function of the population of neurons.2. The method of claim 1 , wherein said population of neurons is hypoxic claim 1 , under oxidative stress claim 1 , under glutamate toxicity or under AMPA/kainate toxicity.3. The method of claim 1 , wherein in step (b) the ratio of the population of astrocytes to neurons is greater than 1:1 claim 1 , 10:1 claim 1 , 100:1 claim 1 , 1000:1 claim 1 , or 10 claim 1 ,000:1.4. The method of claim 1 , wherein said astrocytes express each of GFAP claim 1 , GLAST claim 1 , AQP4 claim 1 , or a combination thereof.5. The method of claim 1 , wherein said astrocytes display secretion of neurotrophic factors selected from the group consisting of BDNF claim 1 , GDNF and VEGF.6. The method of claim 1 , wherein the oxidative stress is selected from the group consisting of reactive oxygen species (ROS) claim 1 , HO claim 1 , and any derivative thereof.7. The method of claim 1 , wherein said quantifying is conducted by counting the number of neurons which are under apoptosis.8. The method of claim 7 , wherein said apoptosis is detected by Caspase-3a labeling claim 7 , Annexin V claim 7 , Tubulin-B3 claim 7 , HB9 or DAPI.9. The method of claim 1 , wherein said agent is a small ...

LIGHT-ACTIVATED CHIMERIC OPSINS AND METHODS OF USING THE SAME

Provided herein are compositions comprising light-activated chimeric proteins expressed on plasma membranes and methods of using the same to selectively depolarize excitatory or inhibitory neurons. 151.-. (canceled)52. A mammalian cell comprising a polynucleotide comprising a nucleotide sequence encoding a light-responsive chimeric polypeptide comprising , in order from N-terminus to C-terminus:a) an amino acid sequence having at least 90% amino acid sequence identity to amino acids 1-163 of the ChR1 amino acid sequence set forth in SEQ ID NO:16; andb) an amino acid sequence having at least 90% amino acid sequence identity to amino acids 120-300 of SEQ ID NO:17.53. The mammalian cell of claim 52 , wherein the light-responsive chimeric polypeptide comprises a C-terminal trafficking signal.54. The mammalian cell of claim 53 , wherein the trafficking signal comprises the amino acid sequence KSRITSEGEYIPLDQIDINV (SEQ ID NO:15).55. The mammalian cell of claim 52 , wherein the nucleotide sequence is operably linked to a promoter.56. The mammalian cell of claim 52 , wherein the light-responsive chimeric polypeptide comprises a Glu to Thr amino acid substitution at position 122 relative to the amino acid sequence set forth in SEQ ID NO:16.57. The mammalian cell of claim 52 , wherein the light-responsive chimeric polypeptide comprises a Glu to Thr amino acid substitution at position 162 relative to the amino acid sequence set forth in SEQ ID NO:16.58. The mammalian cell of claim 52 , wherein the light-responsive chimeric polypeptide comprises a Glu to Thr amino acid substitution at position 122 and a Glu to Thr amino acid substitution at position 162 relative to the amino acid sequence set forth in SEQ ID NO:16.59. The mammalian cell of claim 52 , wherein the cell is a neuronal cell.60. The mammalian cell of claim 52 , wherein the cell is a muscle cell.61. The mammalian cell of claim 52 , wherein the cell is a stem cell.62. The mammalian cell of claim 52 , wherein the cell ...

Control and Characterization of Psychotic States

Provided herein are methods of inducing psychosis in animals using light-responsive opsins and methods of identifying or screening compounds that may be useful in treating psychosis. 112-. (canceled)13. A method of identifying a candidate compound for treating psychosis , the method comprising:a) exposing layer V pyramidal neurons in the prefrontal cortex of a transgenic rat or mouse to light, wherein the genome of the transgenic rat or mouse comprises a nucleic acid sequence encoding a light-responsive depolarizing opsin operably linked to a thymus cell antigen 1 (Thy1) promoter, wherein a subset of layer V pyramidal neurons express on their cell membrane the light-responsive depolarizing opsin, wherein the light-responsive depolarizing opsin comprises an amino acid sequence having at least 90% amino acid sequence identity with the amino acid sequence set forth in one of SEQ ID NOs:1-7, and wherein exposure of the subset of layer V pyramidal neurons to light induces depolarization of the membrane and induces a psychotic state in the transgenic rat or mouse;b) administering the compound to the transgenic rat or mouse having the induced psychotic state; andc) measuring a psychotic state of the transgenic rat or mouse before and after administering the compound, wherein a decrease in the psychotic state indicates that the compound is a candidate for treating psychosis.14. The method of claim 13 , wherein the psychotic state measurement is a behavioral measurement.15. The method of claim 13 , wherein the psychotic state measurement is a cellular measurement.16. The method of claim 13 , further comprising a step of administering a D2 agonist to the animal before administration of the compound.1749-. (canceled)50. The method of claim 14 , wherein the behavioral measurement is social exploration.51. The method of claim 13 , wherein the light-responsive opsin comprises an amino acid sequence having at least 90% identity to the amino acid sequence set forth in SEQ ID NO:1. ...

DIRECT CONVERSION OF CELLS TO CELLS OF OTHER LINEAGES

Methods, compositions and kits for producing functional neurons, astroctyes, oligodendrocytes and progenitor cells thereof are provided. These methods, compositions and kits find use in producing neurons, astrocytes, oligodendrocytes, and progenitor cells thereof for transplantation, for experimental evaluation, as a source of lineage- and cell-specific products, and the like, for example for use in treating human disorders of the CNS. Also provided are methods, compositions and kits for screening candidate agents for activity in converting cells into neuronal cells, astrocytes, oligodendrocytes, and progenitor cells thereof. 1. A method of converting pluripotent cells into induced neuronal cells , the method comprising:contacting a population of pluripotent cells with an effective dose of a neuron reprogramming (NR) system comprising one or more neuron reprogramming (NR) factors, wherein the NR factors are selected from the group consisting of: an Ascl agent, a Ngn agent, and a NeuroD agent for a period of time sufficient to reprogram said pluripotent cells, wherein a population of induced neuronal cells is produced.2. The method of claim 1 , wherein the pluripotent cells are human cells.3. The method of claim 1 , wherein the efficiency of reprogramming said pluripotent cells to become induced neuronal cells is at least about 0.1%.4. The method of claim 1 , wherein the pluripotent cells are reprogrammed into neuronal cells with functional properties as early as 10 days after contacting the cells with the reprogramming factors.5. The method of claim 1 , wherein the pluripotent cells are induced pluripotent (iPS) cells.6. The method of claim 1 , wherein the pluripotent cells are embryonic stem cells.7. The method of claim 1 , wherein the neuron reprogramming factor is Ngn2.8. The method of claim 1 , wherein the induced neuronal cells are excitatory neurons.9. A cell culture system comprising a population of pluripotent cells with an effective dose of a neuron ...

GANGLIOSIDES FOR STANDARDIZING AND INCREASING THE SENSITIVITY OF CELLS TO BOTULINUM NEUROTOXINS IN IN VITRO TEST SYSTEMS

The present invention pertains to a method for standardizing the sensitivity of induced pluripotent stem cell (iPS)-derived neurons to a neurotoxin polypeptide, comprising the steps of: a) cultivating different batches of induced pluripotent stem cell-derived neurons in a cell culture medium comprising GT1b for at least 3 hours; b) contacting the different batches of induced pluripotent stem cell-derived neurons of step a) with a neurotoxin polypeptide; c) cultivating the different batches of induced pluripotent stem cell-derived neurons of step b) for at least 24 hours in the presence of GT1b under conditions which allow for the neurotoxin polypeptide to exert its biological activity, thereby standardizing the sensitivity of the induced pluripotent stem cell-derived neurons to a neurotoxin polypeptide. The invention further relates to a method for the generation of induced pluripotent stem cell-derived neurons having a standardized sensitivity to a neurotoxin polypeptide, comprising the steps of: a) providing different batches of induced pluripotent stem cell-derived neurons; b) cultivating the different batches of induced pluripotent stem cell-derived neurons of step a) in a cell culture medium comprising GT1b for at least 3 hours, thereby standardizing the sensitivity of the induced pluripotent stem cell-derived neurons to a neurotoxin polypeptide. In addition, encompassed by the present invention is a method for determining the biological activity of a neurotoxin polypeptide, comprising the steps of: a) cultivating induced pluripotent stem cell-derived neurons in a cell culture medium comprising GT1b for at least 3 hours; b) contacting the induced pluripotent stem cell-derived neurons of step a) with a neurotoxin polypeptide; c) cultivating the induced pluripotent stem cell-derived neurons of step b) for at least 24 hours in the presence of GT1b under conditions which allow for the neurotoxin polypeptide to exert its biological activity; and d) determining the ...

MODULATION OF SYNAPTIC MAINTENANCE

C1q is shown to be expressed in neurons, where it acts as a signal for synapse elimination. Methods are provided for protecting or treating an individual suffering from adverse effects of synapse loss. These findings have broad implications for a variety of clinical conditions, including Alzheimer's disease. 1. A method of inhibiting synapse loss comprising the step of:administering a therapeutic amount of a complement inhibitor to a patient suffering from adverse synapse loss.2. The method according to claim 1 , wherein said patient has suffered synapse loss as a result of a neurodegenerative disorder.3. The method of claim 2 , wherein said degenerative disorder is a central nervous system disorder.4. The method according to claim 2 , wherein said degenerative disorder is a peripheral nervous system disorder.5. The method of claim 3 , wherein said neurodegenerative disorder is Alzheimer's disease.6. The method according to claim 1 , further comprising administration of neural progenitors claim 1 , or a neurogenesis enhancer.7. The method according to claim 1 , wherein synapse loss is inhibited in said patient.8. The method according to claim 1 , wherein said agent upregulates expression of a complement inhibitor in neural cells.9. The method of claim 1 , wherein said agent inhibits expression of C1q in neural cells.10. The method of claim 1 , wherein said agent inhibits complement activation.11. The method of claim 1 , wherein said agent binds to C1q.12. A method of screening a candidate agent for activity in inhibiting synapse loss claim 1 , the method comprising:contacting a neural cell culture with a candidate agent, wherein said agent is an antagonist or agonist of complement;quantitating the formation of synapses in culture.13. The method of claim 12 , wherein said neural cell culture is a retinal ganglion cell culture.14. The method of claim 12 , wherein said further comprises an astrocyte feeder layer. This invention was made with Government support under ...

Method for screening therapeutic and/or prophylactic agents for alzheimer's disease

The present invention provides a method for screening a therapeutic and/or prophylactic agent for Alzheimer's disease using at least one index selected from the group consisting of the levels of Aβ oligomers, BiP, cleaved caspase 4, PRDX4 and ROS in nerve cells or the like whose differentiation has been induced from iPS cells derived from somatic cells of a patient with Alzheimer's disease.

OLIGONUCLEOTIDES FOR INDUCING PATERNAL UBE3A EXPRESSION

The present invention relates to oligonucleotides that are capable of inducing expression of ubiquitin-protein ligase E3A (UBE3A) from the paternal allele in animal or human neurons. The oligonucleotides target the suppressor of the UBE3A paternal allele by hybridization to SNHG14 long non-coding RNA downstream of SNORD109B. The present invention further relates to pharmaceutical compositions and methods for treatment of Angelman syndrome. 1. An in vivo or in vitro method for inducing UBE3A expression in a target cell where expression of paternal UBE3A is suppressed, said method comprising administering an oligonucleotide consisting of TTAcActtaattatactTCC (SEQ ID NO: 626), wherein capital letters represent beta-D-oxy LNA nucleosides, lowercase letters represent DNA nucleosides, all LNA C are 5-methyl cytosine, and all internucleoside linkages are phosphorothioate internucleoside linkages, in an effective amount to said cell. This application is a continuation and claims priority to application Ser. No. 16/388,714, filed Apr. 18, 2019, which is a continuation and claims priority to application Ser. No. 15/351,113, filed Nov. 14, 2016, which is a continuation and claims priority to PCT/EP2016/077383, filed Nov. 11, 2016, which claims priority to EP15194367.7, filed Nov. 12, 2015 and EP161895024, filed Sep. 19, 2016. The contents of which are hereby incorporated by reference.The present invention relates to oligonucleotides (oligomers) that are complementary to and hybridize to SNHG14 downstream of SNORD109B, leading to induction of paternal expression of Ubiquitin-protein ligase E3A (UBE3A) in an animal or human. The present invention further relates to pharmaceutical compositions and methods for treatment of Angelman syndrome.Angelman syndrome is neuro-genetic disorder caused by deletion or inactivation of the UBE3A genes on the maternally inherited chromosome 15q11.2. The paternal copy of the UBE3A gene is subject to genomic imprinting and silencing in neurons by ...

CELL-BASED METHOD FOR DETERMINING AN ACTIVITY OF BOTULINUM TOXIN

A new cell line and an antibody for determining the activity of botulinum toxin are disclosed. Also disclosed is a method of determining the activity of botulinum toxin using the cell line and/or the antibody. 1. A method for determining activity of botulinum toxin in a sample , comprising the steps of:(a) contacting a cell with the sample, wherein the cell is from a cell line clonally derived from parental neuro-2a cell (accession number KCTC AC28106), wherein the clonally derived cell line has homogenous cell population, and wherein the clonal cell line comprises cells susceptible to intoxication by botulinum toxin type A (BoNT/A) by about 25 pM or less of BoNT/A, and the cell shows higher sensitivity to BoNT/A, BoNT/B, BoNT/C, and BoNT/F compared to the parental neuro-2a cell under same condition, equal sensitivity to BoNT/D of 5 pM or 200 pM concentration to the parental neuro-2a cell under same condition, and no sensitivity to BoNT/E of 10-400 pM concentration;(b) obtaining cell lysate of the cell of (a), said cell lysate comprising proteins of the cell of (a) or isolating proteins from the cell of (a);{'sub': FL', '197, '(c) contacting the cell lysate or the isolated proteins with an agent which specifically binds synaptosomal nerve-associated protein 25 (SNAP25) or botulinum toxin-cleaved SNAP25 fragment (SNAP25);'}{'sub': FL', '197, '(d) detecting the presence of a complex between the agent and the SNAP25and/or SNAP25, and'}{'sub': FL', '197, '(e) determining the activity of botulinum toxin in the sample, wherein the higher the amount of the agent-antigen SNAP25and/or SNAP25complex detected the higher the level of botulinum toxin activity in the sample.'}2. The method of claim 1 , wherein the botulinum toxin is botulinum toxin type A (BoNT/A).3. The method of claim 1 , which further comprises claim 1 , prior to step (a) claim 1 , culturing the cell in a culture medium supplemented with ganglioside GT1b trisodium salt (GT1b).4. The method of claim 3 , wherein ...

NEUROFIBROMATOSES THERAPEUTIC AGENTS AND SCREENING FOR SAME

Disclosed herein are methods of treating a patient at risk of developing or having a neurofibromatosis or a sporadic schwannoma. In exemplary embodiments, the method involves administering to a subject in need an effective amount of a modulator of a target related to neurofibromatosis. Also disclosed are screening assays involving the implementation of Merlin-null Schwann cells, and to compounds identified using same. 1. A method for treating a neurofibromatosis in a subject , the method comprising administering to the subject an effective amount of a NF-related agent.2. The method of claim 1 , wherein the neurofibromatosis is type 1 Neurofibromatosis (NF1).3. The method of claim 1 , wherein the neurofibromatosis is type 2 Neurofibromatosis (NF2).4. The method of claim 1 , wherein the neurofibromatosis is Schwannomatosis.5. The method of claim 1 , wherein the neurofibromatosis includes sporadic Schwannoma in an non-NF patient6. The method of claim 1 , wherein the NF-related agent is selected from the group consisting of a ribozyme claim 1 , an interfering molecule claim 1 , a peptide claim 1 , a small molecule claim 1 , an antibody targeted to LIMK claim 1 , and combinations thereof.7. The method of claim 1 , wherein the NF-related agent comprises a small molecule.8. The method of claim 7 , wherein the NF-related agent comprises a compound from listed in Table 5.9. The method of claim 1 , wherein the NF-related agent comprises an interfering molecule claim 1 , and wherein the interfering molecule comprises a member selected from the group consisting of a phosphothioate morpholino oligomer (PMO) claim 1 , miRNA claim 1 , siRNA claim 1 , methylated siRNA claim 1 , treated siRNAs claim 1 , shRNA claim 1 , antisense RNA claim 1 , a dicer-substrate 27-mer duplex claim 1 , and combinations thereof.10. A method for treating a condition in a subject in need claim 1 , the method comprising administering to the subject an effective amount of a NF-related agent claim 1 , ...

OPTOGENETIC THERAPEUTIC AND METHOD OF TREATING RETINAL DEGENERATIVE AND NEURODEGENERATIVE DISEASES

A method of treating a subject afflicted with a retinal degenerative or neurodegenerative disease including administering an optogenetic therapeutic to the subject, where the optogenetic therapeutic includes an optogenetic protein fused or linked to a marker protein. An optogenetic therapeutic for use in treating a subject afflicted with a retinal degenerative or neurodegenerative disease, including an optogenetic protein fused or linked to a marker protein. 120- (canceled).21. A method of treating a subject afflicted with a retinal degenerative or neurodegenerative disease comprising administering an optogenetic therapeutic to the subject , wherein the optogenetic therapeutic comprises Chronos (ChR90) fused to a marker protein.22. The method of claim 21 , wherein the optogenetic therapeutic is administered to the subject in a therapeutically effective amount capable of treating the retinal degenerative or neurodegenerative disease.23. The method of claim 21 , wherein the marker protein is green fluorescent protein (GFP) claim 21 , enhanced green fluorescent protein (EGFP) claim 21 , yellow fluorescent protein (YFP) claim 21 , or a green fluorescent protein (GFP) derivative.24. The method of claim 21 , wherein the retinal degenerative or neurodegenerative disease is retinitis pigmentosa or macular degeneration.25. The method of claim 21 , wherein the optogenetic therapeutic is administered to retinal output cells of the subject.26. The method of claim 25 , wherein the optogenetic therapeutic is administered via injection.27. The method of claim 21 , further comprising administering a device to the subject claim 21 , wherein the device is configured to take in images via a camera claim 21 , compress and encode the images claim 21 , and transmit coded signals to retinal output cells of the subject.28. The method of claim 27 , wherein the device is embedded in or on a pair of eyeglasses that is worn by the subject. The present application claims priority to U.S. ...

IN VITRO ASSAYS FOR INHIBITION OF MICROGLIAL ACTIVATION

The present invention provides cell-based assays, including high throughput cell-based assays, for identification of candidate therapeutic agents with the ability to inhibit microglial activation in vivo in response to different ligands. 123-. (canceled)24. A cell-based assay for determining toxicity of test agents that inhibit microglia activation in vivo , comprising:providing a composition comprising fibrin and in vitro activated microglial cells;contacting said composition with one or more test agents that exhibit inhibition of in vitro microglia activation; anddetermining toxicity of the identified test agents in vivo by determination of cell death upon exposure of the test agents to the microglial cells.25. The assay of claim 24 , further comprising the step of activating microglial cells in vitro using an agent associated with microglial activation.26. The assay of claim 25 , wherein the microglial cells are activated following introduction of the microglial cells to a fibrin solution.27. The assay of claim 25 , wherein the microglial cells are activated by in vitro exposure to LPS.28. The assay of claim 24 , wherein the test agents exhibit at least a 50% inhibition of the in vitro microglia activation.29. The assay of claim 24 , wherein the cell death is identified through shrinkage of the cell surface area of the microglia.30. The assay of claim 29 , wherein the cell death is identified through shrinkage of the cell surface area of the microglia to 150 μmor less.31. The assay of claim 24 , wherein the test agents comprise agents with demonstrated affinity to targets involved in microglia activation.32. The assay of claim 24 , wherein the test agents comprise agents with no known activity or identified binding target.33. The assay of claim 24 , wherein the test agents comprise known drugs with clinical evaluation data.34. The assay of claim 24 , wherein the test agents comprise known anti-inflammatory and microglia-macrophage response modulators.35. The ...

Methods for labeling and manipulating a cellular circuit

The invention relates to methods and kits for labeling and manipulating a cellular circuit. The methods includes transfecting a first-order cell in the cellular circuit with a nucleic acid molecule encoding a tethered ligand, and a second-order cell in the cellular circuit with a nucleic acid molecule encoding a receptor and an effector fusion polypeptide, a nucleic acid molecule encoding a receptor interactor and protease fusion polypeptide, and a nucleic acid molecule encoding a reporter/modifier gene.

BLOOD BRAIN BARRIER MODELS AND METHODS TO GENERATE AND USE THE SAME

The present disclosure generally relates to a process to prepare a cell culture system that mimics the structure of blood brain barrier (BBB) and are useful to study the functions thereof. In particular, the present invention relates to a direct-contact coculture and triculture systems prepared by plating BMECs on a pre-formed lawn of coculture of astrocytes and pericytes on the apical surface of a culture-chamber to achieve a truly direct contact triculture model for BBB. The cell culture systems disclosed herein are also useful for studying the functions of the blood brain barrier and predicting the efficacy and potential toxicity of a drug candidate. 1. A method for preparing a cell culture system comprising the steps of:g) preparing a cell culture plate with a permeable membrane support;h) seeding a first cell line on said membrane support and proliferating said first cell line for about 2 days in the presence of a cell culture medium;i) removing said cell culture medium and washing proliferated cells of said first cell line;j) seeding a second cell line over proliferated cells of said first cell line;k) proliferating said second cell line and first cell line in the presence of cell culture medium; andl) replacing cell culture medium every other day until proliferated cells reach confluency as determined by stabilized normalized Transendothelial Electrical Resistance (TEER) or by other established methods of assessing cell proliferation or differentiation.2. The method of further comprising a step of: seeding a third cell line over the proliferated cells of said first cell line and proliferating said third cell line for about 2 days in the presence of a culture medium before step d).3. The method of claim 2 , wherein said third cell line is pericytes.4. The method of claim 2 , wherein said first cell line is astrocytes or other glial cells claim 2 , said second cell line is brain microvessel endothelial cells (BMECs) of human or animal origin claim 2 , primary ...

A METHOD FOR PRODUCING AN ADULT-TYPE OLIGODENDROCYTE

The present invention provides a method for producing adult oligodendrocyte progenitor cells from proliferative oligodendrocyte progenitor cells, and a pharmaceutical composition having for an active ingredient thereof adult OPC produced according to that method. The method for producing adult OPC of the present invention is characterized by inducing proliferating OPC to differentiate into adult OPC by culturing in the presence of a ligand of a thyroid hormone receptor or retinoic acid receptor in a low oxygen environment. The present invention further provides adult OPC produced according to the production method of the present invention, and a pharmaceutical composition having these adult OPC as an active ingredient thereof. 2. The method for producing adult OPC according to claim 1 , wherein the ligand is thyroid hormone claim 1 , retinoic acid or vitamin A.3. The method for producing adult OPC according to claim 1 , wherein an oxygen concentration of the low oxygen environment is 0.5% by volume to 1.5% by volume.4. The method for producing adult OPC according to claim 3 , wherein the oxygen concentration is 0.5% by volume to 1.0% by volume.5. The method for producing adult OPC according to claim 1 , wherein the proliferating OPC are cells obtained by primary culturing or sub-culturing OPC harvested from a living body.6. Adult OPC produced by the method for producing adult OPC according to .7. A pharmaceutical composition comprising claim 6 , as an active ingredient thereof claim 6 , the adult OPC according to .8. The pharmaceutical composition according to claim 7 , which is for treatment of hypomyelination claim 7 , demyelination or myelin sheath damage.9. A method for screening for substances having an ability to induce proliferation or induce differentiation in adult OPC claim 7 , comprising:{'claim-ref': {'@idref': 'CLM-00006', 'claim 6'}, 'culturing the adult OPC according to in a medium containing a candidate substance for the ability to induce ...

Compositions to effect neuronal growth

Compositions containing neurogenic agents for inhibition of neuron death and inducing proliferation of neural cells are disclosed.

VITAMIN D RECEPTOR/SMAD GENOMIC CIRCUIT GATES FIBROTIC RESPONSE

The present disclosure provides compositions that include a nanoparticle and a compound that increases the biological activity of the vitamin D receptor (VDR) (e.g., a VDR agonist), and methods of using such compounds to increase retention or storage of vitamin A, vitamin D, and/or lipids by a cell, such as an epithelial or stellate cell. Such methods can be used to treat or prevent fibrosis. 1. A composition , comprising:a nanoparticle comprising on its surface one or more of albumin, retinol binding protein, mannose-6-phosphate modified albumin, a fatty acid ester, a retinyl ester, or a linear-dendritic hybrid polymer; anda compound that increases the biological activity of a vitamin D receptor (VDR) which is in or attached to the nanoparticle.2. The composition of claim 1 , wherein the nanoparticle comprises a lipid nanoparticle or polymeric nanoparticle.3. The composition of claim 1 , wherein the biological activity of the VDR comprises one or more of storage of vitamin A claim 1 , vitamin D and/or lipids by a cell.4. The composition of claim 1 , wherein the compound:increases the biological activity of the VDR by at least 25% as compared to the biological activity in the absence of the compound;increases by at least 25% storage of vitamin A, vitamin D and/or lipids by a cell as compared to the storage in the absence of the compound;increases the biological activity of the VDR in a stellate cell, an epithelial cell, or both;increases the biological activity of the VDR in a pancreatic stellate cell, heart stellate cell, lung stellate cell, kidney stellate cell or hepatic stellate cell;increases by at least 25% storage of vitamin A, vitamin D and/or lipids by a stellate cell as compared to the storage in the absence of the compound; orcombinations thereof.5. The composition of claim 1 , wherein the composition further comprises a chemotherapeutic claim 1 , a biologic claim 1 , or combinations thereof.6. The composition of claim 5 , wherein the chemotherapeutic ...

Method and pharmaceutical composition for treating depression

A method for treating a subject with depression characterized by having an increased burst firing in neurons of a lateral habenula in the subject is provided. The method includes a step of administering to the subject a pharmaceutical composition capable of inhibiting the burst firing in the lateral habenula of the subject. The pharmaceutical composition includes one or more active pharmaceutical agents, which can suppress the burst firing in the lateral habenula of the subject and can include at least one of an N-methyl-D-aspartate receptor (NMDAR) inhibitor or a T-type calcium channel inhibitor. The pharmaceutical composition can be in a formulation allowing for local administration to the lateral habenula of the subject, or can be in a formulation configured for systemic administration to the subject. A method for testing a test substance for an antidepressive effect is also provided.

IN VITRO HUMAN BLOOD BRAIN BARRIER

The present disclosure provides, in some embodiments, in vitro blood brain barrier (iBBB) having functional properties of in vivo BBB as well as methods of identifying compounds capable of traversing the iBBB. Compounds capable of crossing the iBBB and therapeutic uses of such compounds are also described. 1. An in vitro blood brain barrier (iBBB) comprising a 3 dimensional (3D) matrix comprisinga human brain endothelial cell (BEC) vessel comprised of a large interconnected network of human pluripotent-derived positive endothelial cells encapsulated in a 3D matrix,human pluripotent-derived pericytes proximal to the BEC vessel on an apical surface, andhuman pluripotent-derived astrocytes dispersed throughout the 3D matrix, wherein a plurality of the astrocytes are proximal to the BEC vessel and have GFAP-positive projections into the perivascular space.2. The iBBB of claim 1 , wherein the astrocytes express AQP4.3. The iBBB of any one of - claim 1 , wherein the 3D matrix comprises LAMA4.4. The iBBB of any one of - claim 1 , wherein the BEC express at least any one of JAMA claim 1 , PgP claim 1 , LRP1 claim 1 , and RAGE.5. The iBBB of any one of - claim 1 , wherein PgP and ABCG2 are expressed on the apical surface.6. The iBBB of claim 5 , wherein levels of PgP and ABCG2 expressed on the apical surface are 2-3 times greater than levels of PgP and ABCG2 expressed on BEC cultured alone or co-cultured with astrocytes.7. The iBBB of any one of - claim 5 , wherein the iBBB has a TEER that exceeds 5 claim 5 ,500 Ohm×cm2 claim 5 , exhibits reduced molecular permeability and polarization of efflux pumps relative to BEC cultured alone or co-cultured with astrocytes.8. The iBBB of any one of - claim 5 , wherein the iBBB is not cultured with retinoic acid.9. The iBBB of any one of - claim 5 , wherein the human pluripotent are iPSC-derived CD144 cells.10. The iBBB of any one of - claim 5 , wherein the iBBB is generated using 5 parts endothelial cells to 1 part astrocytes to 1 part ...

INSECT-BASED EX VIVO MODEL FOR TESTING BLOOD-BRAIN BARRIER PENETRATION AND METHOD FOR EXPOSING INSECT BRAIN TO NANOPARTICLES

There is provided an ex-vivo insect screening model to accurately determine blood-brain barrier penetration of different nanoparticles in order to improve the compound screening procedures/processes in the early drug discovery process. This object offers many advantages relative to prior technologies since insect models are more reliable tools for the decision-making process than the existing in vitro models, and will speed up the drug screening process and reduce the late phase attrition rate. Moreover, it will reduce the number of mammals sacrificed during the drug discovery phase. 1. A method of conducting blood-brain barrier penetration studies of nanoparticles in an insect , said method comprising the steps:optionally anesthetizing the insect;fixing the head of the insect;dissecting out the dorsal part of the insect head so as to expose the brain, eyes, antennas, and nerve associations;optionally removing the neural lamella;treating the brain with the nanoparticles;washing and homogenising or ultra sound disintegrating the brain; anddetermining the concentration of the nanoparticles in the homogenised brain material.2. The method of claim 1 , wherein the penetration of the nanoparticles through the blood-brain barrier is calculated.3. The method of claim 1 , wherein hemolymph and/or albumin is added to the nanoparticles.4. The method of claim 1 , wherein the concentration of the nanoparticle is determined by LC/MS.5. The method of claim 1 , wherein the brain is treated with the nanoparticles for a period of 1 min to 2 days.6. The method of claim 1 , wherein the neural lamella of the brain are removed before the brain is homogenised or ultra sound disintegrated. The present invention is directed to insect models that are aimed to reflect vertebrate blood-brain barrier (BBB) penetration of nanoparticles. On the other hand BBB penetration of nanoparticles may also cause side effects. Specifically, the present invention relates to the use of insects in screening ...