29,693 results match your criteria vitro describe

The heteromeric complex formed by dopamine receptor D5 and CCR9 leads the gut-homing of CD4 T-cells upon inflammation.

Cell Mol Gastroenterol Hepatol 2021 Apr 14. Epub 2021 Apr 14.

Laboratorio de Neuroinmunología, Fundación Ciencia & Vida, Ñuñoa (7780272), Santiago, Chile; Facultad de Medicina y Ciencia, Universidad San Sebastián, Providencia (7510156), Santiago, Chile. Electronic address:

Background And Aims: CD4 T-cells constitute central players in inflammatory bowel diseases (IBD), driving inflammation in the gut-mucosa. Current evidence indicates that CCR9 and the integrin α4β7 are necessary and sufficient to imprint colonic homing on CD4 T-cells upon inflammation. Interestingly, dopaminergic signalling has been previously involved in leukocyte homing. Read More

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Directed differentiation of human induced pluripotent stem cells to hepatic stellate cells.

Nat Protoc 2021 Apr 16. Epub 2021 Apr 16.

Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), Barcelona, Spain.

Hepatic stellate cells (HSCs) are nonparenchymal liver cells responsible for extracellular matrix homeostasis and are the main cells involved in the development of liver fibrosis following injury. The lack of reliable sources of HSCs has hence limited the development of complex in vitro systems to model liver diseases and toxicity. Here we describe a protocol to differentiate human induced pluripotent stem cells (iPSCs) into hepatic stellate cells (iPSC-HSCs). Read More

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Establishment of caprine airway epithelial cells grown in an air-liquid interface system to study caprine respiratory viruses and bacteria.

Vet Microbiol 2021 Apr 10;257:109067. Epub 2021 Apr 10.

Institute of Virology and Immunology (IVI), Bern, Switzerland; Department of Infectious Diseases and Pathobiology, Vetsuisse Faculty, University of Bern, Bern, Switzerland.

Respiratory diseases negatively impact the global goat industry, but are understudied. There is a shortage of established and biological relevant in vitro or ex vivo assays to study caprine respiratory infections. Here, we describe the establishment of an in vitro system based on well-differentiated caprine airway epithelial cell (AEC) cultures grown under air liquid interface conditions as an experimental platform to study caprine respiratory pathogens. Read More

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Discovery of the First Orally Available, Selective K1.1 Inhibitor: and Activity of an Oxadiazole Series.

ACS Med Chem Lett 2021 Apr 9;12(4):593-602. Epub 2021 Mar 9.

Praxis Precision Medicines, Research Innovation, Cambridge, Massachusetts 02142, United States.

The gene encodes the sodium-activated potassium channel K1.1 (Slack, Slo2.2). Read More

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SOD1 regulates ribosome biogenesis in KRAS mutant non-small cell lung cancer.

Nat Commun 2021 04 15;12(1):2259. Epub 2021 Apr 15.

Rutgers Cancer Institute of New Jersey, Rutgers, The State University of New Jersey, New Brunswick, NJ, USA.

SOD1 is known as the major cytoplasmic superoxide dismutase and an anticancer target. However, the role of SOD1 in cancer is not fully understood. Herein we describe the generation of an inducible Sod1 knockout in KRAS-driven NSCLC mouse model. Read More

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Scaffold-Free Engineering of Human Cartilage Implants.

Cartilage 2021 Apr 15:19476035211007923. Epub 2021 Apr 15.

Department of Immunology, Oslo University Hospital, Oslo, Norway.

Objective: Despite new strategies in tissue engineering, cartilage repair remains a major challenge. Our aim is to treat patients with focal lesions of articular cartilage with autologous hyaline cartilage implants using a scaffold-free approach. In this article, we describe experiments to optimize production of scaffold-free cartilage discs. Read More

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Relating transdermal delivery plasma pharmacokinetics with in vitro permeation test (IVPT) findings using diffusion and compartment-in-series models.

J Control Release 2021 Apr 12. Epub 2021 Apr 12.

Therapeutics Research Group, The University of Queensland Diamantina Institute, The University of Queensland, Woolloongabba, QLD 4102, Australia; Therapeutics Research Centre, University of South Australia Division of Clinical and Health Sciences, Basil Hetzel Institute for Translational Medical Research, The Queen Elizabeth Hospital, Woodville, SA 5011, Australia.

Increasing emphasis is being placed on using in vitro permeation test (IVPT) results for topical products as a surrogate for their in vivo behaviour. This study sought to relate in vivo plasma concentration - time pharmacokinetic (PK) profiles after topical application of drug products to IVPT findings with mechanistic diffusion and compartment models that are now widely used to describe permeation of solutes across the main skin transport barrier, the stratum corneum. Novel in vivo forms of the diffusion and compartment-in-series models were developed by combining their IVPT model forms with appropriate in vivo disposition functions. Read More

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"Photo-Rimonabant": Synthesis and Biological Evaluation of Novel Photoswitchable Molecules Derived from Rimonabant Lead to a Highly Selective and Nanomolar "-On" CBR Antagonist.

ACS Chem Neurosci 2021 Apr 15. Epub 2021 Apr 15.

Pharmazeutische und Medizinische Chemie, Institut für Pharmazie und Lebensmittelchemie, Julius-Maximilians-Universität Würzburg, Am Hubland, D-97074 Würzburg, Germany.

Human cannabinoid receptor type 1 (CBR) plays important roles in the regulation of appetite and development of addictive behaviors. Herein, we describe the design, synthesis, photocharacterization, molecular docking, and characterization of "photo-rimonabant", i.e. Read More

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Intestinal Co-culture System to Study TGR5 Agonism and Gut Restriction.

Bio Protoc 2021 Mar 20;11(6). Epub 2021 Mar 20.

Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, USA.

The activation of the Takeda G-protein receptor 5 (TGR5, also known as the G protein-coupled bile acid receptor 1, GPBAR1) in enteroendocrine L-cells results in secretion of the anti-diabetic hormone Glucagon-Like Peptide 1 (GLP-1) into systemic circulation. Consequently, recent research has focused on identification and development of TGR5 agonists as type 2 diabetes therapeutics. However, the clinical application of TGR5 agonists has been hampered by side effects of these compounds that primarily result from their absorption into circulation. Read More

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Isolation of Extracellular Vesicles Derived from Mesenchymal Stromal Cells by Ultracentrifugation.

Bio Protoc 2020 Dec 20;10(24):e3860. Epub 2020 Dec 20.

Laboratori Experimental de Nefrologia i Trasplantament (LENIT), Institut d'Investigacions Biomèdiques August Pi iSunyer (IDIBAPS), Barcelona, Spain.

Extracellular vesicles (EVs) are a heterogeneous group of membranous vesicles that differ on their biogenesis and release pathways, such as exosomes, microvesicles and apoptotic bodies. They are involved in cell-to-cell communication delivering signal molecules (proteins, nucleic acids, lipids, ) that can regulate different physiological processes, as well as the development and progression of several diseases. There are different methods and commercial kits to isolate EVs and depending on the methodology one could obtain EVs with different degrees of efficiency, purity and it can be more or less time-consuming. Read More

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December 2020

Susceptibility to chlorhexidine and mupirocin among methicillin-resistant Staphylococcus aureus clinical isolates from a teaching hospital.

Rev Inst Med Trop Sao Paulo 2021 12;63:e27. Epub 2021 Apr 12.

Universidade de São Paulo, Faculdade de Medicina, Hospital das Clínicas, São Paulo, São Paulo, Brazil.

Despite the widespread use of chlorhexidine (CHX) to prevent infection, data regarding the in vitro action of CHX against methicillin-resistant Staphylococcus aureus (MRSA) are limited. Clinical isolates from Hospital das Clinicas, Sao Paulo, Brazil, identified during 2002/2003 and 2012/2013 were studied to describe the susceptibility to CHX and mupirocin, molecular characteristics, and virulence profile of MRSA. Susceptibility test to Mupirocin was performed by the disk diffusion method and to CHX by the agar dilution technique. Read More

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Cryopreservation of human cerebral microvascular endothelial cells and astrocytes in suspension and monolayers.

PLoS One 2021 14;16(4):e0249814. Epub 2021 Apr 14.

Department of Chemical and Materials Engineering, University of Alberta, Edmonton, AB, Canada.

The blood-brain barrier (BBB) keeps pathogens and toxins out of the brain but also impedes the entry of pharmaceuticals. Human cerebral microvascular endothelial cells (hCMECs) and astrocytes are the main functional cell components of the BBB. Although available commercially as cryopreserved cells in suspension, improvements in their cryopreservation and distribution as cryopreserved monolayers could enhance BBB in vitro studies. Read More

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Structural basis for broad sarbecovirus neutralization by a human monoclonal antibody.

bioRxiv 2021 Apr 8. Epub 2021 Apr 8.

The recent emergence of SARS-CoV-2 variants of concern (VOC) and the recurrent spillovers of coronaviruses in the human population highlight the need for broadly neutralizing antibodies that are not affected by the ongoing antigenic drift and that can prevent or treat future zoonotic infections. Here, we describe a human monoclonal antibody (mAb), designated S2×259, recognizing a highly conserved cryptic receptor-binding domain (RBD) epitope and cross-reacting with spikes from all sarbecovirus clades. S2×259 broadly neutralizes spike-mediated entry of SARS-CoV-2 including the B. Read More

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Identification of 2,2-Dimethylbutanoic Acid (HST5040), a Clinical Development Candidate for the Treatment of Propionic Acidemia and Methylmalonic Acidemia.

J Med Chem 2021 Apr 13. Epub 2021 Apr 13.

HemoShear Therapeutics Inc., 501 Locust Avenue, Charlottesville, Virginia 22902, United States.

Propionic acidemia (PA) and methylmalonic acidemia (MMA) are rare autosomal recessive disorders of propionyl-CoA (P-CoA) catabolism, caused by a deficiency in the enzymes P-CoA carboxylase and methylmalonyl-CoA (M-CoA) mutase, respectively. PA and MMA are classified as intoxication-type inborn errors of metabolism because the intramitochondrial accumulation of P-CoA, M-CoA, and other metabolites results in secondary inhibition of multiple pathways of intermediary metabolism, leading to organ dysfunction and failure. Herein, we describe the structure-activity relationships of a series of short-chain carboxylic acids which reduce disease-related metabolites in PA and MMA primary hepatocyte disease models. Read More

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Measuring the Complex Effects of the Single-Stranded DNA-Binding Protein gp2.5 on Primer Synthesis and Extension by the Bacteriophage T7 Replisome.

Methods Mol Biol 2021 ;2281:323-332

Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA, USA.

The single-stranded DNA-binding protein gp2.5 of bacteriophage T7 plays myriad functions in the replication of phage genomes. In addition to interacting with ssDNA, gp2. Read More

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January 2021

Stimulation of Variant Forms of the Mitochondrial DNA Helicase Twinkle by the Mitochondrial Single-Stranded DNA-Binding Protein.

Methods Mol Biol 2021 ;2281:313-322

Departamento de Tecnologia, Faculdade de Ciências Agrárias e Veterinárias, Universidade Estadual Paulista "Júlio de Mesquita Filho", Jaboticabal, SP, Brazil.

Defects in mitochondrial DNA (mtDNA) maintenance may lead to disturbances in mitochondrial homeostasis and energy production in eukaryotic cells, causing diseases. During mtDNA replication, the mitochondrial single-stranded DNA-binding protein (mtSSB) stabilizes and protects the exposed single-stranded mtDNA from nucleolysis; perhaps more importantly, it appears to coordinate the actions of both the replicative mtDNA helicase Twinkle and DNA polymerase gamma at the replication fork. Here, we describe a helicase stimulation protocol to test in vitro the functional interaction between mtSSB and variant forms of Twinkle. Read More

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January 2021

Amelioration of Graft-versus-Host Disease by Exopolysaccharide from a Commensal Bacterium.

J Immunol 2021 Apr 12. Epub 2021 Apr 12.

Department of Microbiology and Immunology, Loyola University Chicago, Maywood, IL

Acute graft-versus-host disease (aGvHD) is a severe, often lethal, complication of hematopoietic stem cell transplantation, and although prophylactic regimens are given as standard pretransplantation therapy, up to 60% of these patients develop aGvHD, and require additional immunosuppressive intervention. We treated mice with a purified probiotic molecule, exopolysaccharide (EPS) from shortly before and after induction of aGvHD and found that, whereas only 10% of control mice survived to day 80, 70% of EPS-treated mice survived to 80 d. EPS treatment of donor-only mice resulted in ∼60% survival. Read More

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High-throughput label-free detection of DNA-to-RNA transcription inhibition using brightfield microscopy and deep neural networks.

Comput Biol Med 2021 Apr 4;133:104371. Epub 2021 Apr 4.

Centre de Recherche des Cordeliers, Equipe labellisée par la Ligue contre le Cancer, Université de Paris, Sorbonne Université, Inserm U1138, Institut Universitaire de France, Paris, France; Metabolomics and Cell Biology Platforms, Gustave Roussy Cancer Center, Villejuif, France; Suzhou Institute for Systems Medicine, Chinese Academy of Medical Sciences, Suzhou, China; Po^le de Biologie, Ho^pital Européen Georges Pompidou, AP-HP, Paris, France; Karolinska Institutet, Department of Women's and Children's Health, Karolinska University Hospital, Stockholm, Sweden.

Drug discovery is in constant evolution and major advances have led to the development of in vitro high-throughput technologies, facilitating the rapid assessment of cellular phenotypes. One such phenotype is immunogenic cell death, which occurs partly as a consequence of inhibited RNA synthesis. Automated cell-imaging offers the possibility of combining high-throughput with high-content data acquisition through the simultaneous computation of a multitude of cellular features. Read More

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Culture Methods to Study Apical-Specific Interactions using Intestinal Organoid Models.

J Vis Exp 2021 Mar 23(169). Epub 2021 Mar 23.

STEMCELL Technologies Inc.;

The lining of the gut epithelium is made up of a simple layer of specialized epithelial cells that expose their apical side to the lumen and respond to external cues. Recent optimization of in vitro culture conditions allows for the re-creation of the intestinal stem cell niche and the development of advanced 3-dimensional (3D) culture systems that recapitulate the cell composition and the organization of the epithelium. Intestinal organoids embedded in an extracellular matrix (ECM) can be maintained for long-term and self-organize to generate a well-defined, polarized epithelium that encompasses an internal lumen and an external exposed basal side. Read More

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Biologically derived, 3-dimensional, embryonic scaffolds for long-term cardiomyocyte culture.

Stem Cells Dev 2021 Apr 11. Epub 2021 Apr 11.

University of Minnesota, Internal Medicine/Cardiology, Minneapolis, Minnesota, United States;

The ability to maintain viable cultures of mature, primary cardiomyocytes is challenging. The lack of viable cardiomyocyte cultures severely limits in vitro biochemical assays, toxicology assays, drug screening assays and other analyses. Here, we describe a novel 3-dimensional (3D) embryonic scaffold, which supports the culture of postnatal day 7 (P7) murine cardiomyocytes within the embryonic heart for, at least, 28 days. Read More

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ROS-Induced Hepatotoxicity under Cypermethrin: Involvement of the Crosstalk between Nrf2/Keap1 and NF-κB/iκB-α Pathways Regulated by Proteasome.

Environ Sci Technol 2021 Apr 12. Epub 2021 Apr 12.

College of Wildlife and Protected Area, Northeast Forestry University, Harbin, Heilongjiang 150040, PR China.

Cypermethrin (CMN) is a man-made insecticide, and its abuse has led to potential adverse effects, particularly in sensitive populations such as aquatic organisms. The present study was focused on the toxic phenotype and detoxification mechanism in grass carp () after treatment with waterborne CMN (0.651 μg/L) for 6 weeks in vivo or 6. Read More

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Preclinical development of a molecular clamp-stabilised subunit vaccine for severe acute respiratory syndrome coronavirus 2.

Clin Transl Immunology 2021 5;10(4):e1269. Epub 2021 Apr 5.

School of Chemistry and Molecular Biosciences The University of Queensland St Lucia QLD Australia.

Objectives: Efforts to develop and deploy effective vaccines against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continue at pace. Here, we describe rational antigen design through to manufacturability and vaccine efficacy of a prefusion-stabilised spike (S) protein, Sclamp, in combination with the licensed adjuvant MF59 'MF59C.1' (Seqirus, Parkville, Australia). Read More

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Isolation and culture of adult murine cardiac atrial and ventricular fibroblasts and myofibroblasts.

Methods 2021 Apr 7. Epub 2021 Apr 7.

Montreal Heart Institute, 5000 Belanger, Montréal, Québec, H1T 1C8, Canada; Departments of Biochemistry and Molecular Medicine, Université de Montréal, Montréal, Québec, H3C 3J7, Canada; Pharmacology and Physiology, Université de Montréal, Montréal, Québec, H3C 3J7, Canada; Université de Montréal, Montréal, Québec, H3C 3J7, Canada. Electronic address:

Cardiac fibroblasts play a critical role in extracellular matrix homeostasis, wound healing, and cardiac interstitial fibrosis: the latter being a pathophysiological response to a chronic increase in afterload. Using a standard protocol to isolate cardiac fibroblasts and maintain them in their quiescent phenotype in vitro will enable a better understanding of cardiac fibroblast biology and their role in the response to profibrotic stimuli. Here, we describe an enzymatic method for isolating cardiac fibroblasts. Read More

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In vitro model for predicting the access and distribution of drugs in the brain using hCMEC/D3 cells.

Eur J Pharm Biopharm 2021 Apr 7. Epub 2021 Apr 7.

INEB - Instituto de Engenharia Biomédica, Universidade do Porto, Rua Alfredo Allen 208, 4200-393 Porto, Portugal; i3S - Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Rua Alfredo Allen 208, 4200-393 Porto, Portugal; CESPU - Instituto de Investigação e Formação Avançada em Ciências e Tecnologias da Saúde & Instituto Universitário de Ciências da Saúde, Rua Central de Gandra, 1317, 4585-116 Gandra, Portugal.

The BBB is a protective entity that prevents external substances from reaching the CNS but it also hinders the delivery of drugs into the brain when they are needed. The main objective of this work was to improve a previously proposed in vitro cell-based model by using a more physiological cell line (hCMEC/D3) to predict the main pharmacokinetic parameters that describe the access and distribution of drugs in the CNS: Kp, f, f and V. The hCMEC/D3 permeability of seven drugs was studied in transwell systems under different conditions (standard, modified with albumin and modified with brain homogenate). Read More

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Microglia extracellular traps in Oreochromis niloticus infected with Weissella cibaria.

Fish Shellfish Immunol 2021 Apr 7;113:148-153. Epub 2021 Apr 7.

Department of Technology, School of Agrarian and Veterinary Sciences, Sao Paulo State University (Unesp), 14884-900, Jaboticabal, São Paulo, Brazil.

The mechanism of extracellular traps (ETs) is important in the cellular response against bacteria. Thus, in the present study, we describe for the first time the capacity of the Nile tilapia (Oreochromis niloticus) microglia in the formation of ETs in Weissella cibaria in vitro infection. Thus, we evaluated the ultrastructure of the microglia culture and observed the formation of ETs 6 h after stimulation with lipopolysaccharide (LPS) and during the course of infection. Read More

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Genetic manipulation and immortalized culture of ex vivo primary human germinal center B cells.

Nat Protoc 2021 Apr 9. Epub 2021 Apr 9.

Wellcome-MRC Cambridge Stem Cell Institute, University of Cambridge, Cambridge, UK.

Next-generation sequencing has transformed our knowledge of the genetics of lymphoid malignancies. However, limited experimental systems are available to model the functional effects of these genetic changes and their implications for therapy. The majority of mature B-cell malignancies arise from the germinal center (GC) stage of B-cell differentiation. Read More

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A cell-based screening method using an intracellular antibody for discovering small molecules targeting the translocation protein LMO2.

Sci Adv 2021 Apr 9;7(15). Epub 2021 Apr 9.

Weatherall Institute of Molecular Medicine MRC Molecular Haematology Unit, University of Oxford John Radcliffe Hospital, Oxford OX3 9DS, UK.

Intracellular antibodies are tools that can be used directly for target validation by interfering with properties like protein-protein interactions. An alternative use of intracellular antibodies in drug discovery is developing small-molecule surrogates using antibody-derived (Abd) technology. We previously used this strategy with an in vitro competitive surface plasmon resonance method that relied on high-affinity antibody fragments to obtain RAS-binding compounds. Read More

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Bioengineered3D model of myotonic dystrophy type 1 human skeletal muscle.

Biofabrication 2021 Apr 9. Epub 2021 Apr 9.

Biosensors for Bioengineering, IBEC, c/ Baldiri Reixac 10-12, Parc Científic de Barcelona (PCB), Edifici Clúster, Barcelona, Barcelona, E08028, SPAIN.

Myotonic dystrophy type 1 (DM1) is the most common hereditary myopathy in the adult population. The disease is characterized by progressive skeletal muscle degeneration that produces severe disability. At present, there is still no effective treatment for DM1 patients, but the breakthroughs in understanding the molecular pathogenic mechanisms in DM1 have allowed the testing of new therapeutic strategies. Read More

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Modeling the mature CNS: A predictive screening platform for neurodegenerative disease drug discovery.

J Neurosci Methods 2021 Apr 6:109180. Epub 2021 Apr 6.

Cognition Therapeutics Inc., Pittsburgh, PA, 15203, United States. Electronic address:

Background: Mature primary neuronal cultures are an important model of the nervous system, but limited scalability has been a major challenge in their use for drug discovery of neurodegenerative diseases. This work describes a method for improving scalability through the use of larger format microtiter plates while preserving culture quality.

New Method: Here we describe a method and quality control procedures for growing embryonic day 18 rat hippocampal/cortical neuronal cultures in 384-well microtiter plates for three weeks in vitro. Read More

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Profiling of RNA Structure at Single-Nucleotide Resolution Using nextPARS.

Methods Mol Biol 2021 ;2284:51-61

Barcelona Supercomputing Centre (BSC-CNS), Barcelona, Spain.

RNA molecules play important roles in almost every cellular process, and their functions are mediated by their sequence and structure. Determining the secondary structure of RNAs is central to understanding RNA function and evolution. RNA structure probing techniques coupled to high-throughput sequencing allow determining structural features of RNA molecules at transcriptome-wide scales. Read More

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January 2021