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Article: The Effects of Mechanical Stretch on Integrins and Filopodial-Associated Proteins in Normal and Glaucomatous Trabecular Meshwork Cells.

Yang, Yong-Feng / Sun, Ying Ying / Peters, Donna M / Keller, Kate E

Frontiers in cell and developmental biology

2022 Volume 10, Page(s) 886706

Abstract: The trabecular meshwork (TM) is the tissue responsible for regulating aqueous humor fluid egress from the anterior eye. If drainage is impaired, intraocular pressure (IOP) becomes elevated, which is a primary risk factor for primary open angle glaucoma. ... ...

Abstract	The trabecular meshwork (TM) is the tissue responsible for regulating aqueous humor fluid egress from the anterior eye. If drainage is impaired, intraocular pressure (IOP) becomes elevated, which is a primary risk factor for primary open angle glaucoma. TM cells sense elevated IOP via changes in their biomechanical environment. Filopodia cellular protrusions and integrin transmembrane proteins may play roles in detecting IOP elevation, yet this has not been studied in detail in the TM. Here, we investigate integrins and filopodial proteins, such as myosin-X (Myo10), in response to mechanical stretch, an
Language	English
Publishing date	2022-04-29
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2737824-X
ISSN	2296-634X
ISSN	2296-634X
DOI	10.3389/fcell.2022.886706
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Article ; Online: Digital spatial profiling of segmental outflow regions in trabecular meshwork reveals a role for ADAM15.

Faralli, Jennifer A / Filla, Mark S / Yang, Yong-Feng / Sun, Ying Ying / Johns, Kassidy / Keller, Kate E / Peters, Donna M

PloS one

2024 Volume 19, Issue 2, Page(s) e0298802

Abstract: In this study we used a spatial transcriptomics approach to identify genes specifically associated with either high or low outflow regions in the trabecular meshwork (TM) that could potentially affect aqueous humor outflow in vivo. High and low outflow ... ...

Abstract	In this study we used a spatial transcriptomics approach to identify genes specifically associated with either high or low outflow regions in the trabecular meshwork (TM) that could potentially affect aqueous humor outflow in vivo. High and low outflow regions were identified and isolated from organ cultured human anterior segments perfused with fluorescently-labeled 200 nm FluoSpheres. The NanoString GeoMx Digital Spatial Profiler (DSP) platform was then used to identified genes in the paraffin embedded tissue sections from within those regions. These transcriptome analyses revealed that 16 genes were statistically upregulated in high outflow regions and 57 genes were statistically downregulated in high outflow regions when compared to low outflow regions. Gene ontology enrichment analysis indicated that the top three biological categories of these differentially expressed genes were ECM/cell adhesion, signal transduction, and transcription. The ECM/cell adhesion genes that showed the largest differential expression (Log2FC ±1.5) were ADAM15, BGN, LDB3, and CRKL. ADAM15, which is a metalloproteinase that can bind integrins, was upregulated in high outflow regions, while the proteoglycan BGN and two genes associated with integrin signaling (LDB3, and CRKL) were downregulated. Immunolabeling studies supported the differential expression of ADAM15 and showed that it was specifically upregulated in high outflow regions along the inner wall of Schlemm's canal and in the juxtacanalicular (JCT) region of the TM. In addition to these genes, the studies showed that genes for decorin, a small leucine-rich proteoglycan, and the α8 integrin subunit were enriched in high outflow regions. These studies identify several novel genes that could be involved in segmental outflow, thus demonstrating that digital spatial profiling could be a useful approach for understanding segmental flow through the TM. Furthermore, this study suggests that changes in the expression of genes involved in regulating the activity and/or organization of the ECM and integrins in the TM are likely to be key players in segmental outflow.
MeSH term(s)	Humans ; Trabecular Meshwork/metabolism ; Aqueous Humor/metabolism ; Sclera ; Proteoglycans/metabolism ; Integrins/genetics ; Integrins/metabolism ; Intraocular Pressure ; Membrane Proteins/metabolism ; ADAM Proteins/metabolism
Chemical Substances	Proteoglycans ; Integrins ; ADAM15 protein, human (EC 3.4.24.-) ; Membrane Proteins ; ADAM Proteins (EC 3.4.24.-)
Language	English
Publishing date	2024-02-23
Publishing country	United States
Document type	Journal Article
ZDB-ID	2267670-3
ISSN	1932-6203 ; 1932-6203
ISSN (online)	1932-6203
ISSN	1932-6203
DOI	10.1371/journal.pone.0298802
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Article: Effects of repetitive transcranial magnetic stimulation on motor function and language ability in cerebral palsy: A systematic review and meta-analysis.

Sun, Ying-Ying / Wang, Lei / Peng, Jin-Lin / Huang, Yi-Jie / Qiao, Fu-Qiang / Wang, Pu

Frontiers in pediatrics

2023 Volume 11, Page(s) 835472

Abstract: Objective: This review was conducted to assess the quality of the evidence of effectiveness of repetitive transcranial magnetic stimulation (rTMS) in treating motor and language ability of cerebral palsy (CP).: Method: Medline, Cochrane library, Web ... ...

Abstract	Objective: This review was conducted to assess the quality of the evidence of effectiveness of repetitive transcranial magnetic stimulation (rTMS) in treating motor and language ability of cerebral palsy (CP). Method: Medline, Cochrane library, Web of Science, Embase, PubMed, and CNKI databases were searched up to July 2021 by two independent reviewers. Randomized controlled trials (RCTs) that were published in English and Chinese and met the following criteria were included. The population comprised patients who met the diagnostic criteria for CP. Intervention included the following: comparison about rTMS and sham rTMS or comparison about rTMS combine with other physical therapy and other physical therapy. Outcomes included motor function, as follows: gross motor function measure (GMFM), Gesell Development Diagnosis Scale, fine motor function measure (FMFM), Peabody developmental motor scale, and Modified Ashworth scale. For language ability, sign-significant relation (S-S) was included. Methodological quality was assessed using the Physiotherapy Evidence Database (PEDro) scale. Results: Finally, 29 studies were included in the meta-analysis. Results of evaluation using the Cochrane Collaborative Network Bias Risk Assessment Scale showed that 19 studies specifically explained randomization, among which two studies described allocation concealment, four studies blinded participants and persons and had low risk of bias, and six studies explained that the assessment of outcome measures was blinded. Significant improvements in motor function were observed. The GMFM of total score was determined by using the random-effect model [ Conclusion: The rTMS could improve the motor function and language ability of patients with CP. However, rTMS prescriptions varied, and the studies had low sample sizes. Studies using rigorous and standard research designs about prescriptions and large samples are needed to collect sufficient evidence about the effectiveness of using rTMS to treat patients with CP.
Language	English
Publishing date	2023-02-16
Publishing country	Switzerland
Document type	Journal Article ; Review
ZDB-ID	2711999-3
ISSN	2296-2360
ISSN	2296-2360
DOI	10.3389/fped.2023.835472
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Article: [Analysis of Wumei Pills in treating chronic digestive system diseases with concept of "treating different diseases with same method" based on network pharmacology and molecular docking].

Ding, Jin / Zheng, Pan / Sun, Ying-Ying / Wang, Xiao-Ran / Feng, Yan-Chen

Zhongguo Zhong yao za zhi = Zhongguo zhongyao zazhi = China journal of Chinese materia medica

2022 Volume 47, Issue 15, Page(s) 4164–4176

Abstract: The present study explored the material basis and underlying mechanism of Wumei Pills in the treatment of ulcerative colitis(UC), diabetic enteropathy(DE), and irritable bowel syndrome(IBS) based on network pharmacology and molecular docking.The active ... ...

Abstract	The present study explored the material basis and underlying mechanism of Wumei Pills in the treatment of ulcerative colitis(UC), diabetic enteropathy(DE), and irritable bowel syndrome(IBS) based on network pharmacology and molecular docking.The active components and targets of Wumei Pills were obtained and screened out from TCMSP, and the target names were standardized by UniProt.The related targets of UC, DE, and IBS were searched from GeneCards, DisGeNET, DrugBank, and OMIM.The Venn dia-gram was constructed using the Venny 2.1 online analysis tool to obtain the common targets of the drug and diseases.The "drug-active ingredient-target" network was constructed by Cytoscape 3.7.2.Gene Ontology(GO) function enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analyses of common targets were carried out by DAVID.The main active components and targets were docked by AutoDock.The therapeutic mechanism of Wumei Pills was presumedly related to the regulation of the cancer pathway, TNF signaling pathway, HIF-1 signaling pathway, PI3 K-Akt signaling pathway, NF-κB signaling pathway, Toll-like receptor signaling pathway, JAK-STAT signaling pathway, etc.The results of molecular docking showed that the main active components could bind to the core targets, possessing stable conformation.The therapeutic effects of Wumei Pills against three diseases involved a variety of compounds such as flavonoids, sterols, and alkaloids in the prescriptions, which acted on key targets through multiple organs and participated in multiple signaling pathways such as apoptosis and immune inflammation, thereby exerting the therapeutic action on different diseases with the same method.This study explained the underlying mechanism of Wumei Pills in "treating different diseases with same method", and is expected to provide a theoretical basis for further understanding the mechanism of Wumei Pills and exploring the new clinical application.
MeSH term(s)	Drugs, Chinese Herbal/pharmacology ; Drugs, Chinese Herbal/therapeutic use ; Humans ; Irritable Bowel Syndrome/drug therapy ; Medicine, Chinese Traditional ; Molecular Docking Simulation ; Network Pharmacology
Chemical Substances	Drugs, Chinese Herbal ; wumei
Language	Chinese
Publishing date	2022-08-21
Publishing country	China
Document type	Journal Article
ZDB-ID	1004649-5
ISSN	1001-5302 ; 0254-0029
ISSN	1001-5302 ; 0254-0029
DOI	10.19540/j.cnki.cjcmm.20220412.401
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Article ; Online: Remote ischemic conditioning attenuates oxidative stress and inflammation via the Nrf2/HO-1 pathway in MCAO mice.

Sun, Ying-Ying / Zhu, Hong-Jing / Zhao, Ruo-Yu / Zhou, Sheng-Yu / Wang, Mei-Qi / Yang, Yi / Guo, Zhen-Ni

Redox biology

2023 Volume 66, Page(s) 102852

Abstract: The protective effects of remote ischemic conditioning (RIC) on acute ischemic stroke have been reported. However, the protective mechanisms of RIC have not been fully elucidated. This study aimed to investigate whether RIC could reduce oxidative stress ... ...

Abstract	The protective effects of remote ischemic conditioning (RIC) on acute ischemic stroke have been reported. However, the protective mechanisms of RIC have not been fully elucidated. This study aimed to investigate whether RIC could reduce oxidative stress and inflammatory responses in middle cerebral artery occlusion (MCAO)-reperfusion mice via the nuclear factor-E2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1) pathway. C57BL/6 mice were subjected to MCAO and underwent RIC twice daily at 1, 3, and 7 days after MCAO. ML385 was used to specifically inhibit Nrf2 in MCAO mice. Neurological deficit scores, infarct volume, and hematoxylin-eosin (HE) staining were assessed. Oxidative stress levels were assessed based on total antioxidant capacity (TAC), malonaldehyde (MDA), superoxide dismutase (SOD), and glutathione/glutathione disulfide (GSH/GSSG). mRNA levels were detected using real-time polymerase chain reaction (PCR), and protein levels were detected using western blotting and enzyme-linked immunosorbent assay (ELISA). Protein localization was investigated using immunofluorescence staining. RIC significantly reduced infarct volume and improved neurological function and histological changes after MCAO. RIC significantly increased TAC, SOD, and GSH/GSSG levels and decreased MDA levels. RIC significantly increased Nrf2 and HO-1 mRNA levels and decreased Keap1, NLRP3, and Cleaved Caspase-1 mRNA levels. RIC significantly increased Nrf2, HO-1, and NQO1 protein expression and decreased Keap1, NLRP3, Cleaved Caspase-1, Cleaved IL-1β, IL-6, and TNF-α protein expression. RIC promoted the activation and translocation of Nrf2 into the nucleus. The protective effects of RIC were abolished by ML385 treatment. In conclusion, our findings suggest that RIC alleviates oxidative stress and inflammatory responses via the Nrf2/HO-1 pathway, which in turn improves neurobehavioral function. RIC may provide novel therapeutic options for acute ischemic stroke.
MeSH term(s)	Animals ; Mice ; Mice, Inbred C57BL ; Kelch-Like ECH-Associated Protein 1 ; NF-E2-Related Factor 2/genetics ; Infarction, Middle Cerebral Artery ; Ischemic Stroke ; Heme Oxygenase-1/genetics ; Glutathione Disulfide ; NLR Family, Pyrin Domain-Containing 3 Protein ; Oxidative Stress ; Antioxidants ; Inflammation ; Caspase 1
Chemical Substances	Kelch-Like ECH-Associated Protein 1 ; NF-E2-Related Factor 2 ; Heme Oxygenase-1 (EC 1.14.14.18) ; Glutathione Disulfide (ULW86O013H) ; NLR Family, Pyrin Domain-Containing 3 Protein ; Antioxidants ; Caspase 1 (EC 3.4.22.36)
Language	English
Publishing date	2023-08-16
Publishing country	Netherlands
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2701011-9
ISSN	2213-2317 ; 2213-2317
ISSN (online)	2213-2317
ISSN	2213-2317
DOI	10.1016/j.redox.2023.102852
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Article: Screening and isolation of glyceroglycolipids with antialgal activity from several marine macroalgae

Sun, Ying-Ying / Dong, Sha-Sha / Zhang, Nai-Sheng / Zhou, Jing / Long, Zheng-Kang

Journal of applied phycology. 2021 Aug., v. 33, no. 4

2021

Abstract: Glyceroglycolipids and antialgal activity were used as the main indexes to identify the glyceroglycolipid extracts of marine macroalgae with significant antialgal activity from 6 species of marine macroalgae—Codium fragile, Neoporphyra haitanensis, ... ...

Abstract	Glyceroglycolipids and antialgal activity were used as the main indexes to identify the glyceroglycolipid extracts of marine macroalgae with significant antialgal activity from 6 species of marine macroalgae—Codium fragile, Neoporphyra haitanensis, Sargassum fusiforme, Saccharina japonica, Silvetia siliquosa and Undaria pinnatifida. The extracts of all tested algae contained glyceroglycolipids and exhibited antialgal activity against Karenia mikimotoi and Phaeocystis globosa. Further, by antialgal activity bioassay-guided fractionation method in combination with extraction and partitioning as well as multi-chromatography, two glyceroglycolipids with antialgal activity, 1-O-palmitoyl-3-O-β-D-galactopyranosyl glycerol and 1-O-palmitoyl-2-O-oleoyl-3-O-β-D-galactopyranosyl glycerol, were isolated for the first time from N. haitanensis and S. siliquosa, respectively. There were no glyceroglycolipids with antialgal activity in the other four macroalgae.
Keywords	Karenia mikimotoi ; Phaeocystis globosa ; Saccharina japonica ; Sargassum fusiforme ; Silvetia ; Undaria pinnatifida ; algology ; fractionation ; glycerol ; macroalgae
Language	English
Dates of publication	2021-08
Size	p. 2609-2616.
Publishing place	Springer Netherlands
Document type	Article
ZDB-ID	1002324-0
ISSN	1573-5176 ; 0921-8971
ISSN (online)	1573-5176
ISSN	0921-8971
DOI	10.1007/s10811-021-02466-4
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Gentulizumab, a novel anti-CD47 antibody with potent antitumor activity and demonstrates a favorable safety profile.

Wang, Tao / Wang, Si-Qin / Du, Yin-Xiao / Sun, Dan-Dan / Liu, Chang / Liu, Shuang / Sun, Ying-Ying / Wang, Hai-Long / Zhang, Chun-Sheng / Liu, Hai-Long / Jin, Lei / Chen, Xiao-Ping

Journal of translational medicine

2024 Volume 22, Issue 1, Page(s) 220

Abstract: Background: Targeting CD47/SIRPα axis has emerged as a promising strategy in cancer immunotherapy. Despite the encouraging clinical efficacy observed in hematologic malignancies through CD47-SIRPα blockade, there are safety concerns related to the ... ...

Abstract	Background: Targeting CD47/SIRPα axis has emerged as a promising strategy in cancer immunotherapy. Despite the encouraging clinical efficacy observed in hematologic malignancies through CD47-SIRPα blockade, there are safety concerns related to the binding of anti-CD47 antibodies to CD47 on the membrane of peripheral blood cells. Methods: In order to enhance the selectivity and therapeutic efficacy of the antibody, we developed a humanized anti-CD47 monoclonal antibody called Gentulizumab (GenSci059). The binding capacity of GenSci059 to CD47 was evaluated using flow cytometry and surface plasmon resonance (SPR) methods, the inhibitory effect of GenSci059 on the CD47-SIRPα interaction was evaluated through competitive ELISA assays. The anti-tumor activity of GenSci059 was assessed using in vitro macrophage models and in vivo patient-derived xenograft (PDX) models. To evaluate the safety profile of GenSci059, binding assays were conducted using blood cells. Additionally, we investigated the underlying mechanisms contributing to the weaker binding of GenSci059 to erythrocytes. Finally, toxicity studies were performed in non-human primates to assess the potential risks associated with GenSci059. Results: GenSci059 displayed strong binding to CD47 in both human and monkey, and effectively inhibited the CD47-SIRPα interaction. With doses ranging from 5 to 20 mg/kg, GenSci059 demonstrated potent inhibition of the growth of subcutaneous tumor with the inhibition rates ranged from 30.3% to complete regression. Combination of GenSci059 with 2.5 mg/kg Rituximab at a dose of 2.5 mg/kg showed enhanced tumor inhibition compared to monotherapy, exhibiting synergistic effects. GenSci059 exhibited minimal binding to hRBCs compared to Hu5F9-G4. The binding of GenSci059 to CD47 depended on the cyclization of N-terminal pyroglutamic acid and the spatial conformation of CD47, but was not affected by its glycosylation modifications. A maximum tolerated dose (MTD) of 450 mg/kg was observed for GenSci059, and no significant adverse effects were observed in repeated dosages up to 10 + 300 mg/kg, indicating a favorable safety profile. Conclusion: GenSci059 selectively binds to CD47, effectively blocks the CD47/SIRPα axis signaling pathway and enhances the phagocytosis effects of macrophages toward tumor cells. This monoclonal antibody demonstrates potent antitumor activity and exhibits a favorable safety profile, positioning it as a promising and effective therapeutic option for cancer.
MeSH term(s)	Animals ; Humans ; CD47 Antigen ; Neoplasms/pathology ; Phagocytosis ; Macrophages/metabolism ; Antibodies, Monoclonal/pharmacology ; Antibodies, Monoclonal/therapeutic use ; Immunotherapy/methods ; Disease Models, Animal ; Antigens, Differentiation/metabolism ; Antigens, Differentiation/pharmacology ; Antigens, Differentiation/therapeutic use
Chemical Substances	CD47 Antigen ; Antibodies, Monoclonal ; Antigens, Differentiation ; CD47 protein, human
Language	English
Publishing date	2024-03-01
Publishing country	England
Document type	Journal Article
ZDB-ID	2118570-0
ISSN	1479-5876 ; 1479-5876
ISSN (online)	1479-5876
ISSN	1479-5876
DOI	10.1186/s12967-023-04710-6
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Article: Nanodrug delivery systems for regulating microglial polarization in ischemic stroke treatment: A review.

Lei, Shuang-Yin / Yang, Yu-Qian / Liu, Jia-Cheng / Zhang, Dian-Hui / Qu, Yang / Sun, Ying-Ying / Zhu, Hong-Jing / Zhou, Sheng-Yu / Yang, Yi / Guo, Zhen-Ni

Journal of tissue engineering

2024 Volume 15, Page(s) 20417314241237052

Abstract: The incidence of ischemic stroke (IS) is rising in tandem with the global aging population. There is an urgent need to delve deeper into the pathological mechanisms and develop new neuroprotective strategies. In the present review, we discuss the latest ... ...

Abstract	The incidence of ischemic stroke (IS) is rising in tandem with the global aging population. There is an urgent need to delve deeper into the pathological mechanisms and develop new neuroprotective strategies. In the present review, we discuss the latest advancements and research on various nanodrug delivery systems (NDDSs) for targeting microglial polarization in IS treatment. Furthermore, we critically discuss the different strategies. NDDSs have demonstrated exceptional qualities to effectively permeate the blood-brain barrier, aggregate at the site of ischemic injury, and target specific cell types within the brain when appropriately modified. Consequently, NDDSs have considerable potential for reshaping the polarization phenotype of microglia and could be a prospective therapeutic strategy for IS. The treatment of IS remains a challenge. However, this review provides a new perspective on neuro-nanomedicine for IS therapies centered on microglial polarization, thereby inspiring new research ideas and directions.
Language	English
Publishing date	2024-03-13
Publishing country	England
Document type	Journal Article ; Review
ZDB-ID	2573915-3
ISSN	2041-7314
ISSN	2041-7314
DOI	10.1177/20417314241237052
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Article ; Online: Phenotypic and Functional Alterations in Tunneling Nanotubes Formed by Glaucomatous Trabecular Meshwork Cells.

Sun, Ying Ying / Bradley, John M / Keller, Kate E

Investigative ophthalmology & visual science

2019 Volume 60, Issue 14, Page(s) 4583–4595

Abstract: Purpose: Trabecular meshwork (TM) cells detect and coordinate responses to intraocular pressure (IOP) in the eye. TM cells become dysfunctional in glaucoma where IOP is often elevated. Recently, we showed that normal TM (NTM) cells communicate by ... ...

Abstract	Purpose: Trabecular meshwork (TM) cells detect and coordinate responses to intraocular pressure (IOP) in the eye. TM cells become dysfunctional in glaucoma where IOP is often elevated. Recently, we showed that normal TM (NTM) cells communicate by forming tubular connections called tunneling nanotubes (TNTs). Here, we investigated TNTs in glaucomatous TM (GTM) cells. Methods: Primary GTM and NTM cells were established from cadaver eyes. Transfer of Vybrant DiO and DiD-labeled vesicles via TNT connections was measured. Imaris software measured the number and length of cell protrusions from immunofluorescent confocal images. Live-cell imaging of the actin cytoskeleton was performed. The distribution of myosin-X, a regulator of TNTs/filopodia, was investigated in TM cells and tissue. Results: GTM cells contained significantly more transferred fluorescent vesicles than NTM cells (49.6% vs. 35%). Although NTM cells had more protrusions at the cell surface than GTM cells (7.61 vs. 4.65 protrusions/cell), GTM protrusions were significantly longer (12.1 μm vs. 9.76 μm). Live-cell imaging demonstrated that the GTM actin cytoskeleton was less dynamic, and vesicle transfer between cells was significantly slower than NTM cells. Furthermore, rearrangement of the actin cortex adjacent to the TNT may influence TNT formation. Myosin-X immunostaining was punctate and disorganized in GTM cells and tissue compared to age-matched NTM controls. Conclusions: Together, our data demonstrate that GTM cells have phenotypic and functional differences in their TNTs. Significantly slower vesicle transfer via TNTs in GTM cells may delay the timely propagation of cellular signals when pressures become elevated in glaucoma.
MeSH term(s)	Actin Cytoskeleton/metabolism ; Blotting, Western ; Cell Size ; Cells, Cultured ; Cellular Senescence/physiology ; Densitometry ; Glaucoma, Open-Angle/metabolism ; Glaucoma, Open-Angle/pathology ; Humans ; Microscopy, Confocal ; Myosins/metabolism ; Nanotubes ; Phagocytosis/physiology ; Phenotype ; Pseudopodia/metabolism ; Signal Transduction/physiology ; Trabecular Meshwork/metabolism ; Trabecular Meshwork/pathology
Chemical Substances	MYO10 protein, human ; Myosins (EC 3.6.4.1)
Language	English
Publishing date	2019-11-17
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	391794-0
ISSN	1552-5783 ; 0146-0404
ISSN (online)	1552-5783
ISSN	0146-0404
DOI	10.1167/iovs.19-28084
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Article ; Online: Myosin-X Silencing in the Trabecular Meshwork Suggests a Role for Tunneling Nanotubes in Outflow Regulation.

Sun, Ying Ying / Yang, Yong-Feng / Keller, Kate E

Investigative ophthalmology & visual science

2019 Volume 60, Issue 2, Page(s) 843–851

Abstract: Purpose: The actin cytoskeleton plays a key role in outflow regulation through the trabecular meshwork (TM). Although actin stress fibers are a target of glaucoma therapies, the role of other actin cellular structures is unclear. Myosin-X (Myo10) is an ... ...

Abstract	Purpose: The actin cytoskeleton plays a key role in outflow regulation through the trabecular meshwork (TM). Although actin stress fibers are a target of glaucoma therapies, the role of other actin cellular structures is unclear. Myosin-X (Myo10) is an actin-binding protein that is involved in tunneling nanotube (TNT) and filopodia formation. Here, we inhibited Myo10 pharmacologically or by gene silencing to investigate the role of filopodia/TNTs in the TM. Methods: Short hairpin RNA interference (RNAi) silencing lentivirus targeting myosin-X (shMyo10) was generated. Human anterior segments were perfused with shMyo10 or CK-666, an Arp2/3 inhibitor. Confocal microscopy investigated the colocalization of Myo10 with matrix metalloproteinase (MMPs). Western immunoblotting investigated the protein levels of MMPs and extracellular matrix (ECM) proteins. MMP activity and phagocytosis assays were performed. Results: CK-666 and shMyo10-silencing lentivirus caused a significant reduction in outflow rates in anterior segment perfusion culture, an ex vivo method to study intraocular pressure regulation. In human TM cells, Myo10 colocalized with MMP2, MMP14, and cortactin in podosome-like structures, which function as regions of focal ECM degradation. Furthermore, MMP activity, thrombospondin-1 and SPARC protein levels were significantly reduced in the media of CK-666-treated and shMyo10-silenced TM cells. However, neither Myo10 silencing or CK-666 treatment significantly affected phagocytic uptake. Conclusions: Inhibiting filopodia/TNTs caused opposite effects on outflow compared with inhibiting stress fibers. Moreover, Myo10 may also play a role in focal ECM degradation in TM cells. Our results provide additional insight into the function of actin supramolecular assemblies and actin-binding proteins in outflow regulation.
MeSH term(s)	Aged ; Aged, 80 and over ; Aqueous Humor/physiology ; Blotting, Western ; Extracellular Matrix Proteins/metabolism ; Female ; Fluorescent Antibody Technique, Indirect ; Gene Silencing ; Humans ; Indoles/pharmacology ; Lentivirus/genetics ; Male ; Matrix Metalloproteinases/metabolism ; Microscopy, Confocal ; Microvessels/physiology ; Middle Aged ; Myosins/physiology ; Nanotubes ; Phagocytosis ; Pseudopodia/drug effects ; Pseudopodia/metabolism ; RNA Interference/physiology ; Real-Time Polymerase Chain Reaction ; Trabecular Meshwork/metabolism
Chemical Substances	CK-0944666 ; Extracellular Matrix Proteins ; Indoles ; MYO10 protein, human ; Matrix Metalloproteinases (EC 3.4.24.-) ; Myosins (EC 3.6.4.1)
Language	English
Publishing date	2019-02-26
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	391794-0
ISSN	1552-5783 ; 0146-0404
ISSN (online)	1552-5783
ISSN	0146-0404
DOI	10.1167/iovs.18-26055
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