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Article ; Online: Stereoselective Synthesis of the Spirocyclic γ-Lactam Core of the Ansalactams.

Liang, Zhanhao / Lin, You-Chen / Pierce, Joshua G

Organic letters

2021 Volume 23, Issue 24, Page(s) 9559–9562

Abstract: Ansalactam A is an ansa macrolide natural product that contains a densely functionalized spiro-γ-lactam core containing three contiguous stereocenters. This unusual motif distinguishes it from other members of this family and represents a significant ... ...

Abstract	Ansalactam A is an ansa macrolide natural product that contains a densely functionalized spiro-γ-lactam core containing three contiguous stereocenters. This unusual motif distinguishes it from other members of this family and represents a significant synthetic challenge. Herein, we report the development of a stereoselective formal [3+2] cycloaddition reaction for the construction of this key spiro-γ-lactam motif for the first time, thereby enabling access to the northern domain of ansalactam A.
MeSH term(s)	Spiro Compounds
Chemical Substances	Spiro Compounds
Language	English
Publishing date	2021-11-30
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ISSN	1523-7052
ISSN (online)	1523-7052
DOI	10.1021/acs.orglett.1c03782
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Differential Expression Analysis Identifies Candidate Synaptogenic Molecules for Wiring Direction-Selective Circuits in the Retina.

Tworig, Joshua M / Morrie, Ryan D / Bistrong, Karina / Somaiya, Rachana D / Hsu, Shaw / Liang, Jocelyn / Cornejo, Karen G / Feller, Marla B

The Journal of neuroscience : the official journal of the Society for Neuroscience

2024 Volume 44, Issue 18

Abstract: An organizational feature of neural circuits is the specificity of synaptic connections. A striking example is the direction-selective (DS) circuit of the retina. There are multiple subtypes of DS retinal ganglion cells (DSGCs) that prefer motion along ... ...

Abstract	An organizational feature of neural circuits is the specificity of synaptic connections. A striking example is the direction-selective (DS) circuit of the retina. There are multiple subtypes of DS retinal ganglion cells (DSGCs) that prefer motion along one of four preferred directions. This computation is mediated by selective wiring of a single inhibitory interneuron, the starburst amacrine cell (SAC), with each DSGC subtype preferentially receiving input from a subset of SAC processes. We hypothesize that the molecular basis of this wiring is mediated in part by unique expression profiles of DSGC subtypes. To test this, we first performed paired recordings from isolated mouse retinas of both sexes to determine that postnatal day 10 (P10) represents the age at which asymmetric synapses form. Second, we performed RNA sequencing and differential expression analysis on isolated P10 ON-OFF DSGCs tuned for either nasal or ventral motion and identified candidates which may promote direction-specific wiring. We then used a conditional knock-out strategy to test the role of one candidate, the secreted synaptic organizer cerebellin-4 (Cbln4), in the development of DS tuning. Using two-photon calcium imaging, we observed a small deficit in directional tuning among ventral-preferring DSGCs lacking Cbln4, though whole-cell voltage-clamp recordings did not identify a significant change in inhibitory inputs. This suggests that Cbln4 does not function primarily via a cell-autonomous mechanism to instruct wiring of DS circuits. Nevertheless, our transcriptomic analysis identified unique candidate factors for gaining insights into the molecular mechanisms that instruct wiring specificity in the DS circuit.
MeSH term(s)	Animals ; Mice ; Retina/metabolism ; Retina/physiology ; Male ; Synapses/physiology ; Synapses/metabolism ; Female ; Retinal Ganglion Cells/metabolism ; Retinal Ganglion Cells/physiology ; Mice, Inbred C57BL ; Amacrine Cells/physiology ; Amacrine Cells/metabolism ; Motion Perception/physiology ; Nerve Net/physiology ; Nerve Net/metabolism ; Visual Pathways/physiology ; Visual Pathways/metabolism
Language	English
Publishing date	2024-05-01
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	604637-x
ISSN	1529-2401 ; 0270-6474
ISSN (online)	1529-2401
ISSN	0270-6474
DOI	10.1523/JNEUROSCI.1461-23.2024
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Zs.A 1668: Show issues

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Article: Stereoselective Synthesis of the Spirocyclic γ-Lactam Core of the Ansalactams

Liang, Zhanhao / Lin, You-Chen / Pierce, Joshua G.

Organic letters. 2021 Nov. 30, v. 23, no. 24

2021

Abstract	Ansalactam A is an ansa macrolide natural product that contains a densely functionalized spiro-γ-lactam core containing three contiguous stereocenters. This unusual motif distinguishes it from other members of this family and represents a significant synthetic challenge. Herein, we report the development of a stereoselective formal [3+2] cycloaddition reaction for the construction of this key spiro-γ-lactam motif for the first time, thereby enabling access to the northern domain of ansalactam A.
Keywords	cycloaddition reactions ; macrolides ; stereoselective synthesis ; stereoselectivity
Language	English
Dates of publication	2021-1130
Size	p. 9559-9562.
Publishing place	American Chemical Society
Document type	Article
ISSN	1523-7052
DOI	10.1021/acs.orglett.1c03782
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Article ; Online: Perinuclear positioning of endosomes can affect PS-ASO activities.

Liang, Xue-Hai / Nichols, Joshua G / Tejera, Dario / Crooke, Stanley T

Nucleic acids research

2022 Volume 49, Issue 22, Page(s) 12970–12985

Abstract: Phosphorothioate (PS) modified antisense oligonucleotide (ASO) drugs that act on cellular RNAs must enter cells and be released from endocytic organelles to elicit antisense activity. It has been shown that PS-ASOs are mainly released by late endosomes. ... ...

Abstract	Phosphorothioate (PS) modified antisense oligonucleotide (ASO) drugs that act on cellular RNAs must enter cells and be released from endocytic organelles to elicit antisense activity. It has been shown that PS-ASOs are mainly released by late endosomes. However, it is unclear how endosome movement in cells contributes to PS-ASO activity. Here, we show that PS-ASOs in early endosomes display Brownian type motion and migrate only short distances, whereas PS-ASOs in late endosomes (LEs) move linearly along microtubules with substantial distances. In cells with normal microtubules and LE movement, PS-ASO-loaded LEs tend to congregate perinuclearly. Disruption of perinuclear positioning of LEs by reduction of dynein 1 decreased PS-ASO activity, without affecting PS-ASO cellular uptake. Similarly, disruption of perinuclear positioning of PS-ASO-LE foci by reduction of ER tethering proteins RNF26, SQSTM1 and UBE2J1, or by overexpression of P50 all decreased PS-ASO activity. However, enhancing perinuclear positioning through reduction of USP15 or over-expression of RNF26 modestly increased PS-ASO activity, indicating that LE perinuclear positioning is required for ensuring efficient PS-ASO release. Together, these observations suggest that LE movement along microtubules and perinuclear positioning affect PS-ASO productive release.
MeSH term(s)	Animals ; Biological Transport ; Cell Line, Tumor ; Cell Nucleus/metabolism ; Cells, Cultured ; Dyneins/metabolism ; Endoplasmic Reticulum/metabolism ; Endosomes/metabolism ; HeLa Cells ; Humans ; Mice ; Microscopy, Confocal ; Microtubules/metabolism ; Motion ; Neoplasm Proteins/metabolism ; Oligonucleotides, Antisense/genetics ; Oligonucleotides, Antisense/metabolism ; Thionucleotides/genetics ; Thionucleotides/metabolism
Chemical Substances	Neoplasm Proteins ; Oligonucleotides, Antisense ; RNF26 protein, human ; Thionucleotides ; Dyneins (EC 3.6.4.2)
Language	English
Publishing date	2022-01-10
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	186809-3
ISSN	1362-4962 ; 1362-4954 ; 0301-5610 ; 0305-1048
ISSN (online)	1362-4962 ; 1362-4954
ISSN	0301-5610 ; 0305-1048
DOI	10.1093/nar/gkab1198
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Zs.A 1067: Show issues

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Article ; Online: Hsc70 Facilitates Mannose-6-Phosphate Receptor-Mediated Intracellular Trafficking and Enhances Endosomal Release of Phosphorothioate-Modified Antisense Oligonucleotides.

Liang, Xue-Hai / Nichols, Joshua G / Hsu, Chih-Wei / Crooke, Stanley T

Nucleic acid therapeutics

2021 Volume 31, Issue 4, Page(s) 284–297

Abstract: Phosphorothioate-modified antisense oligonucleotide (PS-ASO) drugs are commonly used to modulate gene expression through RNase H1-mediated cleavage of target RNAs. Upon internalization through endocytic pathways into cells, PS-ASOs must be released from ... ...

Abstract	Phosphorothioate-modified antisense oligonucleotide (PS-ASO) drugs are commonly used to modulate gene expression through RNase H1-mediated cleavage of target RNAs. Upon internalization through endocytic pathways into cells, PS-ASOs must be released from membraned endosomal organelles to act on target RNAs, a limiting step of PS-ASO activity. Here we report that Hsc70 protein mediates productive release of PS-ASOs from endosomes. Hsc70 protein was enriched in endosome fractions shortly after PS-ASO incubation with cells. Reduction of Hsc70 significantly decreased the activities of PS-ASOs in reducing target RNAs. PS-ASO uptake and transport from early endosomes to late endosomes (LEs) were not affected upon Hsc70 reduction; however, endosomal release of PS-ASOs was impaired. Reduction of Hsc70 led to more scattered mannose-6-phosphate receptor (M6PR) localization at LEs in the cytoplasm, in contrast to the perinuclear localization at
MeSH term(s)	Endocytosis ; Endosomes ; Oligonucleotides, Antisense/genetics ; Phosphorothioate Oligonucleotides ; Receptor, IGF Type 2/genetics
Chemical Substances	Oligonucleotides, Antisense ; Phosphorothioate Oligonucleotides ; Receptor, IGF Type 2
Language	English
Publishing date	2021-02-09
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2639888-6
ISSN	2159-3345 ; 2159-3337
ISSN (online)	2159-3345
ISSN	2159-3337
DOI	10.1089/nat.2020.0920
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Article ; Online: The mitochondrial ATP synthase is a negative regulator of the mitochondrial permeability transition pore.

Pekson, Ryan / Liang, Felix G / Axelrod, Joshua L / Lee, Jaehoon / Qin, Dongze / Wittig, Andre J H / Paulino, Victor M / Zheng, Min / Peixoto, Pablo M / Kitsis, Richard N

Proceedings of the National Academy of Sciences of the United States of America

2023 Volume 120, Issue 51, Page(s) e2303713120

Abstract: The mitochondrial permeability transition pore (mPTP) is a channel in the inner mitochondrial membrane whose sustained opening in response to elevated mitochondrial matrix ... ...

Abstract	The mitochondrial permeability transition pore (mPTP) is a channel in the inner mitochondrial membrane whose sustained opening in response to elevated mitochondrial matrix Ca
MeSH term(s)	Mice ; Animals ; Mitochondrial Proton-Translocating ATPases/genetics ; Mitochondrial Proton-Translocating ATPases/metabolism ; Mitochondrial Permeability Transition Pore/metabolism ; Mitochondrial Membrane Transport Proteins/metabolism ; Cyclophilins/genetics ; Cyclophilins/metabolism ; Peptidyl-Prolyl Isomerase F ; Mitochondria, Heart/genetics ; Mitochondria, Heart/metabolism ; Calcium/metabolism
Chemical Substances	Mitochondrial Proton-Translocating ATPases (EC 3.6.3.-) ; Mitochondrial Permeability Transition Pore ; Mitochondrial Membrane Transport Proteins ; Cyclophilins (EC 5.2.1.-) ; Peptidyl-Prolyl Isomerase F ; Calcium (SY7Q814VUP)
Language	English
Publishing date	2023-12-13
Publishing country	United States
Document type	Journal Article
ZDB-ID	209104-5
ISSN	1091-6490 ; 0027-8424
ISSN (online)	1091-6490
ISSN	0027-8424
DOI	10.1073/pnas.2303713120
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Zs.A 1148: Show issues

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Article ; Online: Author Correction: Circulating and urinary tumour DNA in urothelial carcinoma - upper tract, lower tract and metastatic disease.

Rose, Kyle M / Huelster, Heather L / Meeks, Joshua J / Faltas, Bishoy M / Sonpavde, Guru P / Lerner, Seth P / Ross, Jeffrey S / Spiess, Philippe E / Grass, G Daniel / Jain, Rohit K / Kamat, Ashish M / Vosoughi, Aram / Wang, Liang / Wang, Xuefeng / Li, Roger

Nature reviews. Urology

2023 Volume 20, Issue 7, Page(s) 452

Language	English
Publishing date	2023-05-23
Publishing country	England
Document type	Published Erratum
ZDB-ID	2493737-X
ISSN	1759-4820 ; 1759-4812
ISSN (online)	1759-4820
ISSN	1759-4812
DOI	10.1038/s41585-023-00783-6
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Zs.A 6030: Show issues

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Article ; Online: Ageing causes an aortic contractile dysfunction phenotype by targeting the expression of members of the extracellular signal-regulated kinase pathway.

Nicholson, Christopher J / Xing, Yi / Lee, Sophie / Liang, Stephanie / Mohan, Shivani / O'Rourke, Caitlin / Kang, Joshua / Morgan, Kathleen G

Journal of cellular and molecular medicine

2022 Volume 26, Issue 5, Page(s) 1456–1465

Abstract: The extracellular signal-regulated kinase (ERK) pathway is a well-known regulator of vascular smooth muscle cell proliferation, but it also serves as a regulator of caldesmon, which negatively regulates vascular contractility. This study examined whether ...

Abstract	The extracellular signal-regulated kinase (ERK) pathway is a well-known regulator of vascular smooth muscle cell proliferation, but it also serves as a regulator of caldesmon, which negatively regulates vascular contractility. This study examined whether aortic contractile function requires ERK activation and if this activation is regulated by ageing. Biomechanical experiments revealed that contractile responses to the alpha1-adrenergic agonist phenylephrine are attenuated specifically in aged mice, which is associated with downregulation of ERK phosphorylation. ERK inhibition attenuates phenylephrine-induced contractility, indicating that the contractile tone is at least partially ERK-dependent. To explore the mechanisms of this age-related downregulation of ERK phosphorylation, we transfected microRNAs, miR-34a and miR-137 we have previously shown to increase with ageing and demonstrated that in A7r5 cells, both miRs downregulate the expression of Src and paxillin, known regulators of ERK signalling, as well as ERK phosphorylation. Further studies in aortic tissues transfected with miRs show that miR-34a but not miR-137 has a negative effect on mRNA levels of Src and paxillin. Furthermore, ERK phosphorylation is decreased in aortic tissue treated with the Src inhibitor PP2. Increases in miR-34a and miR-137 with ageing downregulate the expression of Src and paxillin, leading to impaired ERK signalling and aortic contractile dysfunction.
MeSH term(s)	Aging/genetics ; Animals ; Extracellular Signal-Regulated MAP Kinases/metabolism ; Mice ; MicroRNAs/genetics ; MicroRNAs/metabolism ; Paxillin/genetics ; Paxillin/metabolism ; Phenotype ; Phenylephrine/pharmacology ; Phosphorylation
Chemical Substances	MicroRNAs ; Paxillin ; Phenylephrine (1WS297W6MV) ; Extracellular Signal-Regulated MAP Kinases (EC 2.7.11.24)
Language	English
Publishing date	2022-02-18
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2074559-X
ISSN	1582-4934 ; 1582-4934 ; 1582-1838
ISSN (online)	1582-4934
ISSN	1582-4934 ; 1582-1838
DOI	10.1111/jcmm.17118
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Zs.A 5752: Show issues

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Article ; Online: Pulmonary Vascular Dilatation Detected by Automated Transcranial Doppler in COVID-19 Pneumonia.

Reynolds, Alexandra S / Lee, Alison G / Renz, Joshua / DeSantis, Katherine / Liang, John / Powell, Charles A / Ventetuolo, Corey E / Poor, Hooman D

American journal of respiratory and critical care medicine

2020 Volume 202, Issue 7, Page(s) 1037–1039

Keywords	covid19
Language	English
Publishing date	2020-09-09
Publishing country	United States
Document type	Letter ; Research Support, N.I.H., Extramural
ZDB-ID	1180953-x
ISSN	1535-4970 ; 0003-0805 ; 1073-449X
ISSN (online)	1535-4970
ISSN	0003-0805 ; 1073-449X
DOI	10.1164/rccm.202006-2219LE
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Uh II Zs.80: Show issues

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Article ; Online: Data-Dependent Acquisition with Precursor Coisolation Improves Proteome Coverage and Measurement Throughput for Label-Free Single-Cell Proteomics.

Truong, Thy / Webber, Kei G I / Madisyn Johnston, S / Boekweg, Hannah / Lindgren, Caleb M / Liang, Yiran / Nydegger, Alissia / Xie, Xiaofeng / Tsang, Tsz-Ming / Jayatunge, D A Dasun N / Andersen, Joshua L / Payne, Samuel H / Kelly, Ryan T

Angewandte Chemie (International ed. in English)

2023 Volume 62, Issue 34, Page(s) e202303415

Abstract: We combined efficient sample preparation and ultra-low-flow liquid chromatography with a newly developed data acquisition and analysis scheme termed wide window acquisition (WWA) to quantify >3,000 proteins from single cells in rapid label-free analyses. ...

Abstract	We combined efficient sample preparation and ultra-low-flow liquid chromatography with a newly developed data acquisition and analysis scheme termed wide window acquisition (WWA) to quantify >3,000 proteins from single cells in rapid label-free analyses. WWA employs large isolation windows to intentionally co-isolate and co-fragment adjacent precursors along with the selected precursor. Optimized WWA increased the number of MS2-identified proteins by ≈40 % relative to standard data-dependent acquisition. For a 40-min LC gradient operated at ≈15 nL/min, we identified an average of 3,524 proteins per single-cell-sized aliquot of protein digest. Reducing the active gradient to 20 min resulted in a modest 10 % decrease in proteome coverage. Using this platform, we compared protein expression between single HeLa cells having an essential autophagy gene, atg9a, knocked out, with their isogenic WT parental line. Similar proteome coverage was observed, and 268 proteins were significantly up- or downregulated. Protein upregulation primarily related to innate immunity, vesicle trafficking and protein degradation.
MeSH term(s)	Humans ; Proteome/analysis ; HeLa Cells ; Proteomics/methods ; Chromatography, Liquid/methods
Chemical Substances	Proteome
Language	English
Publishing date	2023-07-13
Publishing country	Germany
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	2011836-3
ISSN	1521-3773 ; 1433-7851
ISSN (online)	1521-3773
ISSN	1433-7851
DOI	10.1002/anie.202303415
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