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Article ; Online: PC-PLC/sphingomyelin synthase activity plays a central role in the development of myogenic tone in murine resistance arteries.

Mauban, Joseph R H / Zacharia, Joseph / Fairfax, Seth / Wier, Withrow Gil

American journal of physiology. Heart and circulatory physiology

2015 Volume 308, Issue 12, Page(s) H1517–24

Abstract: Myogenic tone is an intrinsic property of the vasculature that contributes to blood pressure control and tissue perfusion. Earlier investigations assigned a key role in myogenic tone to phospholipase C (PLC) and its products, inositol 1,4,5-trisphosphate ...

Abstract	Myogenic tone is an intrinsic property of the vasculature that contributes to blood pressure control and tissue perfusion. Earlier investigations assigned a key role in myogenic tone to phospholipase C (PLC) and its products, inositol 1,4,5-trisphosphate (IP3) and diacylglycerol (DAG). Here, we used the PLC inhibitor, U-73122, and two other, specific inhibitors of PLC subtypes (PI-PLC and PC-PLC) to delineate the role of PLC in myogenic tone of pressurized murine mesenteric arteries. U-73122 inhibited depolarization-induced contractions (high external K(+) concentration), thus confirming reports of nonspecific actions of U-73122 and its limited utility for studies of myogenic tone. Edelfosine, a specific inhibitor of PI-PLC, did not affect depolarization-induced contractions but modulated myogenic tone. Because PI-PLC produces IP3, we investigated the effect of blocking IP3 receptor-mediated Ca(2+) release on myogenic tone. Incubation of arteries with xestospongin C did not affect tone, consistent with the virtual absence of Ca(2+) waves in arteries with myogenic tone. D-609, an inhibitor of PC-PLC and sphingomyelin synthase, strongly inhibited myogenic tone and had no effect on depolarization-induced contraction. D-609 appeared to act by lowering cytoplasmic Ca(2+) concentration to levels below those that activate contraction. Importantly, incubation of pressurized arteries with a membrane-permeable analog of DAG induced vasoconstriction. The results therefore mandate a reexamination of the signaling pathways activated by the Bayliss mechanism. Our results suggest that PI-PLC and IP3 are not required in maintaining myogenic tone, but DAG, produced by PC-PLC and/or SM synthase, is likely through multiple mechanisms to increase Ca(2+) entry and promote vasoconstriction.
MeSH term(s)	Animals ; Arterial Pressure ; Calcium Signaling/drug effects ; Diglycerides/metabolism ; Enzyme Inhibitors/pharmacology ; Mesenteric Arteries/enzymology ; Mice ; Muscle, Smooth, Vascular/drug effects ; Muscle, Smooth, Vascular/enzymology ; Phosphatidylcholines/metabolism ; Signal Transduction/drug effects ; Time Factors ; Transferases (Other Substituted Phosphate Groups)/antagonists & inhibitors ; Transferases (Other Substituted Phosphate Groups)/metabolism ; Type C Phospholipases/antagonists & inhibitors ; Type C Phospholipases/metabolism ; Vascular Resistance/drug effects ; Vasoconstriction/drug effects
Chemical Substances	Diglycerides ; Enzyme Inhibitors ; Phosphatidylcholines ; Transferases (Other Substituted Phosphate Groups) (EC 2.7.8.-) ; phosphatidylcholine-ceramide phosphocholine transferase (EC 2.7.8.-) ; Type C Phospholipases (EC 3.1.4.-) ; phosphatidylcholine-specific phospholipase C (EC 3.1.4.3)
Language	English
Publishing date	2015-04-17
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	603838-4
ISSN	1522-1539 ; 0363-6135
ISSN (online)	1522-1539
ISSN	0363-6135
DOI	10.1152/ajpheart.00594.2014
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Article ; Online: Vascular tone and Ca(2+) signaling in murine cremaster muscle arterioles in vivo.

Mauban, Joseph R H / Zacharia, Joseph / Zhang, Jin / Wier, Withrow Gil

Microcirculation (New York, N.Y. : 1994)

2012 Volume 20, Issue 3, Page(s) 269–277

Abstract: ... which is not due to α1 -AR, AT1 R, or SNA. PLC activity, L-type Ca(2+) channels, 2-APB- and SKF96365 ... sensitive channels are required. Propagating Ca(2+) waves are not present. A key role for PLC and InsP3 R ...

Abstract	Objectives: We sought to determine some of the molecular requirements for basal state "tone" of skeletal muscle arterioles in vivo, and whether asynchronous Ca(2+) waves are involved or not. Methods: Cremaster muscles of anesthetized exMLCK and smGCaMP2 biosensor mice were exteriorized, and the fluorescent arterioles were visualized with wide-field, confocal or multiphoton microscopy to observe Ca(2+) signaling and arteriolar diameter. Results: Basal state tone of the arterioles was ~50%. Local block of Ang-II receptors (AT1 ) or α1 -adrenoceptors (α1 -AR) had no effect on diameter, nor did complete block of sympathetic nerve activity (SNA). Inhibition of phospholipase C caused dilation nearly to the Ca(2+) -free (passive) diameter, as did exposure to nifedipine or 2-APB. Arterioles were also dilated when treated with SKF96365. High-resolution imaging of exMLCK fluorescence (ratio) or GCaMP2 fluorescence in smooth muscle cells failed to reveal Ca(2+) waves (although Ca(2+) waves/transients were readily detected by both biosensors in small arteries, ex vivo). Conclusions: Arterioles of cremaster muscle have vascular tone of ~ 50%, which is not due to α1 -AR, AT1 R, or SNA. PLC activity, L-type Ca(2+) channels, 2-APB- and SKF96365-sensitive channels are required. Propagating Ca(2+) waves are not present. A key role for PLC and InsP3 R in vascular tone in vivo, other than producing Ca(2+) waves, is suggested.
MeSH term(s)	Animals ; Arterioles/cytology ; Arterioles/metabolism ; Calcium/metabolism ; Calcium Channels/metabolism ; Calcium Signaling/physiology ; Mice ; Mice, Transgenic ; Microscopy, Fluorescence, Multiphoton ; Muscle, Skeletal/blood supply ; Muscle, Skeletal/cytology
Chemical Substances	Calcium Channels ; Calcium (SY7Q814VUP)
Language	English
Publishing date	2012-11-12
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	1217758-1
ISSN	1549-8719 ; 1073-9688
ISSN (online)	1549-8719
ISSN	1073-9688
DOI	10.1111/micc.12025
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Article ; Online: A method for noninvasive longitudinal measurements of [Ca2+] in arterioles of hypertensive optical biosensor mice.

Mauban, Joseph R H / Fairfax, Seth T / Rizzo, Mark A / Zhang, Jin / Wier, Withrow Gil

American journal of physiology. Heart and circulatory physiology

2014 Volume 307, Issue 2, Page(s) H173–81

Abstract: We used two-photon (2-p) Förster resonance energy transfer (FRET) microscopy to provide serial, noninvasive measurements of [Ca(2+)] in arterioles of living "biosensor" mice. These express a genetically encoded Ca(2+) indicator (GECI), either FRET-based ... ...

Abstract	We used two-photon (2-p) Förster resonance energy transfer (FRET) microscopy to provide serial, noninvasive measurements of [Ca(2+)] in arterioles of living "biosensor" mice. These express a genetically encoded Ca(2+) indicator (GECI), either FRET-based exMLCK or intensity-based GCaMP2. The FRET ratios, Rmin and Rmax, required for in vivo Ca(2+) calibration of exMLCK were obtained in isolated arteries. For in vivo experiments, mice were anesthetized (1.5% isoflurane), and arterioles within a depilated ear were visualized through the intact skin (i.e., noninvasively), by 2-p excitation of exMLCK (at 820 nm) or GCaMP2 (at 920 nm). Spontaneous or agonist-evoked [Ca(2+)] transients in arteriolar smooth muscle cells were imaged (at 2 Hz) with both exMLCK and GCaMP2. To examine changes in arteriolar [Ca(2+)] that might accompany hypertension, five exMLCK mice were implanted with telemetric blood pressure transducers and osmotic minipumps containing ANG II (350 ng·kg(-1)·min(-1)) and fed a high (6%)-salt diet for 9 days. [Ca(2+)] was measured every other day in five smooth muscle cells of two to three arterioles in each animal. Prior to ANG II/salt, [Ca(2+)] was 246 ± 42 nM. [Ca(2+)] increased transiently to 599 nM on day 2 after beginning ANG II/salt, then remained elevated at 331 ± 42 nM for 4 more days, before returning to 265 ± 47 nM 6 days after removal of ANG II/salt. In summary, two-photon excitation of exMLCK and GCaMP2 provides a method for noninvasive, longitudinal quantification of [Ca(2+)] dynamics and vascular structure in individual arterioles of a particular animal over an extended period of time, a capability that should enhance future studies of hypertension and vascular function.
MeSH term(s)	Angiotensin II ; Animals ; Arterial Pressure ; Arterioles/metabolism ; Arterioles/physiopathology ; Biosensing Techniques ; Calcium/metabolism ; Calcium Signaling ; Disease Models, Animal ; Fluorescence Resonance Energy Transfer ; Hypertension/chemically induced ; Hypertension/metabolism ; Hypertension/physiopathology ; Mice ; Mice, Transgenic ; Microscopy, Fluorescence, Multiphoton ; Muscle, Smooth, Vascular/metabolism ; Muscle, Smooth, Vascular/physiopathology ; Myosin-Light-Chain Kinase/genetics ; Myosin-Light-Chain Kinase/metabolism ; Skin/blood supply ; Sodium Chloride, Dietary ; Time Factors
Chemical Substances	Sodium Chloride, Dietary ; Angiotensin II (11128-99-7) ; Myosin-Light-Chain Kinase (EC 2.7.11.18) ; Calcium (SY7Q814VUP)
Language	English
Publishing date	2014-05-23
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	603838-4
ISSN	1522-1539 ; 0363-6135
ISSN (online)	1522-1539
ISSN	0363-6135
DOI	10.1152/ajpheart.00182.2014
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Article ; Online: High vascular tone of mouse femoral arteries in vivo is determined by sympathetic nerve activity via α1A- and α1D-adrenoceptor subtypes.

Zacharia, Joseph / Mauban, Joseph R H / Raina, Hema / Fisher, Steven A / Wier, Withrow G

PloS one

2013 Volume 8, Issue 6, Page(s) e65969

Abstract: Background and purpose: Determining the role of vascular receptors in vivo is difficult and not readily accomplished by systemic application of antagonists or genetic manipulations. Here we used intravital microscopy to measure the contributions of ... ...

Abstract	Background and purpose: Determining the role of vascular receptors in vivo is difficult and not readily accomplished by systemic application of antagonists or genetic manipulations. Here we used intravital microscopy to measure the contributions of sympathetic receptors, particularly α1-adrenoceptor subtypes, to contractile activation of femoral artery in vivo. Experimental approach: Diameter and intracellular calcium ([Ca(2+)]i) in femoral arteries were determined by intravital fluorescence microscopy in mice expressing a Myosin Light Chain Kinase (MLCK) based calcium-calmodulin biosensor. Pharmacological agents were applied locally to the femoral artery to determine the contributions of vascular receptors to tonic contraction and [Ca(2+)]i,. Key results: In the anesthetized animal, femoral arteries were constricted to a diameter equal to 54% of their passive diameter (i.e. tone = 46%). Of this total basal tone, 16% was blocked by RS79948 (0.1 µM) and thus attributable to α2-adrenoceptors. A further 46% was blocked by prazosin (0.1 µM) and thus attributable to α1-adrenoceptors. Blockade of P2X and NPY1 receptors with suramin (0.5 mM) and BIBP3226 (1.0 µM) respectively, reduced tone by a further 22%, leaving 16% of basal tone unaffected at these concentrations of antagonists. Application of RS100329 (α1A-selective antagonist) and BMY7378 (α1D-selective) decreased tone by 29% and 26%, respectively, and reduced [Ca(2+)]i. Chloroethylclonidine (1 µM preferential for α1B-) had no effect. Abolition of sympathetic nerve activity (hexamethonium, i.p.) reduced basal tone by 90%. Conclusion and implications: Tone of mouse femoral arteries in vivo is almost entirely sympathetic in origin. Activation of α1A- and α1D-adrenoceptors elevates [Ca(2+)]i and accounts for at least 55% of the tone.
MeSH term(s)	Adrenergic Fibers/physiology ; Adrenergic alpha-1 Receptor Antagonists/pharmacology ; Animals ; Arginine/analogs & derivatives ; Calcium/metabolism ; Femoral Artery/innervation ; Femoral Artery/physiology ; Isoquinolines ; Mice ; Microscopy, Fluorescence ; Muscle Tonus/drug effects ; Muscle Tonus/physiology ; Muscle, Smooth, Vascular/physiology ; Myography ; Myosin-Light-Chain Kinase ; Naphthyridines ; Piperazines ; Prazosin ; Receptors, Adrenergic, alpha-1/metabolism ; Suramin ; Thymine
Chemical Substances	Adrenergic alpha-1 Receptor Antagonists ; BIBP 3226 ; Isoquinolines ; Naphthyridines ; Piperazines ; RS 100329 ; RS 79948-197 ; Receptors, Adrenergic, alpha-1 ; Suramin (6032D45BEM) ; Arginine (94ZLA3W45F) ; Myosin-Light-Chain Kinase (EC 2.7.11.18) ; BMY 7378 (KC07KV8T5O) ; Thymine (QR26YLT7LT) ; Calcium (SY7Q814VUP) ; Prazosin (XM03YJ541D)
Language	English
Publishing date	2013-06-12
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ISSN	1932-6203
ISSN (online)	1932-6203
DOI	10.1371/journal.pone.0065969
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Article ; Online: Non-specific binding and steric hindrance thresholds for penetration of particulate drug carriers within tumor tissue.

Dancy, Jimena G / Wadajkar, Aniket S / Schneider, Craig S / Mauban, Joseph R H / Goloubeva, Olga G / Woodworth, Graeme F / Winkles, Jeffrey A / Kim, Anthony J

Journal of controlled release : official journal of the Controlled Release Society

2016 Volume 238, Page(s) 139–148

Abstract: Therapeutic nanoparticles (NPs) approved for clinical use in solid tumor therapy provide only modest improvements in patient survival, in part due to physiological barriers that limit delivery of the particles throughout the entire tumor. Here, we ... ...

Abstract	Therapeutic nanoparticles (NPs) approved for clinical use in solid tumor therapy provide only modest improvements in patient survival, in part due to physiological barriers that limit delivery of the particles throughout the entire tumor. Here, we explore the thresholds for NP size and surface poly(ethylene glycol) (PEG) density for penetration within tumor tissue extracellular matrix (ECM). We found that NPs as large as 62nm, but less than 110nm in diameter, diffused rapidly within a tumor ECM preparation (Matrigel) and breast tumor xenograft slices ex vivo. Studies of PEG-density revealed that increasing PEG density enhanced NP diffusion and that PEG density below a critical value led to adhesion of NP to ECM. Non-specific binding of NPs to tumor ECM components was assessed by surface plasmon resonance (SPR), which revealed excellent correlation with the particle diffusion results. Intravital microscopy of NP spread in breast tumor tissue confirmed a significant difference in tumor tissue penetration between the 62 and 110nm PEG-coated NPs, as well as between PEG-coated and uncoated NPs. SPR assays also revealed that Abraxane, an FDA-approved non-PEGylated NP formulation used for cancer therapy, binds to tumor ECM. Our results establish limitations on the size and surface PEG density parameters required to achieve uniform and broad dispersion within tumor tissue and highlight the utility of SPR as a high throughput method to screen NPs for tumor penetration.
MeSH term(s)	Albumin-Bound Paclitaxel/administration & dosage ; Albumin-Bound Paclitaxel/metabolism ; Animals ; Antineoplastic Agents/administration & dosage ; Antineoplastic Agents/metabolism ; Breast/drug effects ; Breast/metabolism ; Breast Neoplasms/drug therapy ; Breast Neoplasms/metabolism ; Cell Line, Tumor ; Collagen/metabolism ; Diffusion ; Doxorubicin/administration & dosage ; Doxorubicin/analogs & derivatives ; Doxorubicin/metabolism ; Drug Carriers/analysis ; Drug Carriers/metabolism ; Drug Combinations ; Female ; Humans ; Lactic Acid/analysis ; Lactic Acid/metabolism ; Laminin/metabolism ; Mice ; Mice, Nude ; Nanoparticles/analysis ; Nanoparticles/metabolism ; Neoplasms/drug therapy ; Neoplasms/metabolism ; Particle Size ; Polyethylene Glycols/administration & dosage ; Polyethylene Glycols/analysis ; Polyethylene Glycols/metabolism ; Polyglycolic Acid/analysis ; Polyglycolic Acid/metabolism ; Polylactic Acid-Polyglycolic Acid Copolymer ; Proteoglycans/metabolism ; Surface Properties
Chemical Substances	Albumin-Bound Paclitaxel ; Antineoplastic Agents ; Drug Carriers ; Drug Combinations ; Laminin ; Proteoglycans ; liposomal doxorubicin ; matrigel (119978-18-6) ; Polylactic Acid-Polyglycolic Acid Copolymer (1SIA8062RS) ; Polyglycolic Acid (26009-03-0) ; Lactic Acid (33X04XA5AT) ; Polyethylene Glycols (3WJQ0SDW1A) ; Doxorubicin (80168379AG) ; Collagen (9007-34-5)
Language	English
Publishing date	2016-07-25
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	632533-6
ISSN	1873-4995 ; 0168-3659
ISSN (online)	1873-4995
ISSN	0168-3659
DOI	10.1016/j.jconrel.2016.07.034
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Zs.A 2055: Show issues

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Article: Ca(2+) signaling in arterioles and small arteries of conscious, restrained, optical biosensor mice.

Fairfax, Seth T / Mauban, Joseph R H / Hao, Scarlett / Rizzo, Mark A / Zhang, Jin / Wier, W Gil

Frontiers in physiology

2014 Volume 5, Page(s) 387

Abstract: Unlabelled: Two-photon fluorescence microscopy and conscious, restrained optical biosensor mice were used to study smooth muscle Ca(2+) signaling in ear arterioles. Conscious mice were used in order to preserve normal mean arterial blood pressure (MAP) ... ...

Abstract	Unlabelled: Two-photon fluorescence microscopy and conscious, restrained optical biosensor mice were used to study smooth muscle Ca(2+) signaling in ear arterioles. Conscious mice were used in order to preserve normal mean arterial blood pressure (MAP) and sympathetic nerve activity (SNA). ExMLCK mice, which express a genetically-encoded smooth muscle-specific FRET-based Ca(2+) indicator, were equipped with blood pressure telemetry and immobilized for imaging. MAP was 101 ± 4 mmHg in conscious restrained mice, similar to the freely mobile state (107 ± 3 mmHg). Oscillatory vasomotion or irregular contractions were observed in most arterioles (71%), with the greatest oscillatory frequency observed at 0.25 s(-1). In a typical arteriole with an average diameter of ~35 μm, oscillatory vasomotion of a 5-6 μm magnitude was accompanied by nearly uniform [Ca(2+)] oscillations from ~0.1 to 0.5 μM, with maximum [Ca(2+)] occurring immediately before the rapid decrease in diameter. Very rapid, spatially uniform "Ca(2+) flashes" were also observed but not asynchronous propagating Ca(2+) waves. In contrast, vasomotion and dynamic Ca(2+) signals were rarely observed in ear arterioles of anesthetized exMLCK biosensor mice. Hexamethonium (30 μg/g BW, i.p.) caused a fall in MAP to 74 ± 4 mmHg, arteriolar vasodilation, and abolition of vasomotion and synchronous Ca(2+) transients. Summary: MAP and heart rate (HR) were normal during high-resolution Ca(2+) imaging of conscious, restrained mice. SNA induced continuous vasomotion and irregular vasoconstrictions via spatially uniform Ca(2+) signaling within the arterial wall. FRET-based biosensor mice and two-photon imaging provided the first measurements of [Ca(2+)] in vascular smooth muscle cells in arterioles of conscious animals.
Language	English
Publishing date	2014-10-07
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2564217-0
ISSN	1664-042X
ISSN	1664-042X
DOI	10.3389/fphys.2014.00387
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Article: AKAP-scaffolding proteins and regulation of cardiac physiology.

Mauban, J R H / O'Donnell, M / Warrier, S / Manni, S / Bond, M

Physiology (Bethesda, Md.)

2009 Volume 24, Page(s) 78–87

Abstract: A kinase anchoring proteins (AKAPs) compose a growing list of diverse but functionally related proteins defined by their ability to bind to the regulatory subunit of protein kinase A. AKAPs perform an integral role in the spatiotemporal modulation of a ... ...

Abstract	A kinase anchoring proteins (AKAPs) compose a growing list of diverse but functionally related proteins defined by their ability to bind to the regulatory subunit of protein kinase A. AKAPs perform an integral role in the spatiotemporal modulation of a multitude of cellular signaling pathways. This review highlights the extensive role of AKAPs in cardiac excitation/contraction coupling and cardiac physiology. The literature shows that particular AKAPs are involved in cardiac Ca(2+) influx, release, reuptake, and myocyte repolarization. Studies have also suggested roles for AKAPs in cardiac remodeling. Transgenic studies show functional effects of AKAPs, not only in the cardiovascular system but in other organ systems as well.
MeSH term(s)	A Kinase Anchor Proteins/physiology ; Animals ; Dimerization ; Heart/physiology ; Humans ; Mice ; Mice, Knockout ; Myocardium/enzymology
Chemical Substances	A Kinase Anchor Proteins
Language	English
Publishing date	2009-04-13
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	2158667-6
ISSN	1548-9221 ; 1548-9213
ISSN (online)	1548-9221
ISSN	1548-9213
DOI	10.1152/physiol.00041.2008
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Zs.A 2164: Show issues

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Article: Hypoxic pulmonary vasoconstriction: role of ion channels.

Mauban, Joseph R H / Remillard, Carmelle V / Yuan, Jason X-J

Journal of applied physiology (Bethesda, Md. : 1985)

2005 Volume 98, Issue 1, Page(s) 415–420

Abstract: Acute hypoxia induces pulmonary vasoconstriction and chronic hypoxia causes structural changes of the pulmonary vasculature including arterial medial hypertrophy. Electro- and pharmacomechanical mechanisms are involved in regulating pulmonary vasomotor ... ...

Abstract	Acute hypoxia induces pulmonary vasoconstriction and chronic hypoxia causes structural changes of the pulmonary vasculature including arterial medial hypertrophy. Electro- and pharmacomechanical mechanisms are involved in regulating pulmonary vasomotor tone, whereas intracellular Ca(2+) serves as an important signal in regulating contraction and proliferation of pulmonary artery smooth muscle cells. Herein, we provide a basic overview of the cellular mechanisms involved in the development of hypoxic pulmonary vasoconstriction. Our discussion focuses on the roles of ion channels permeable to K(+) and Ca(2+), membrane potential, and cytoplasmic Ca(2+) in the development of acute hypoxic pulmonary vasoconstriction and chronic hypoxia-mediated pulmonary vascular remodeling.
MeSH term(s)	Animals ; Calcium/metabolism ; Hemostasis ; Humans ; Hypertension, Pulmonary/etiology ; Hypertension, Pulmonary/physiopathology ; Hypoxia/complications ; Hypoxia/physiopathology ; Ion Channel Gating ; Ion Channels/metabolism ; Lung/blood supply ; Lung/physiopathology ; Models, Biological ; Oxygen/metabolism ; Pulmonary Artery/physiopathology ; Vasoconstriction
Chemical Substances	Ion Channels ; Oxygen (S88TT14065) ; Calcium (SY7Q814VUP)
Language	English
Publishing date	2005-01
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, P.H.S. ; Review
ZDB-ID	219139-8
ISSN	1522-1601 ; 8750-7587 ; 0021-8987 ; 0161-7567
ISSN (online)	1522-1601
ISSN	8750-7587 ; 0021-8987 ; 0161-7567
DOI	10.1152/japplphysiol.00732.2004
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Article: Histamine-mediated increases in cytosolic [Ca2+] involve different mechanisms in human pulmonary artery smooth muscle and endothelial cells.

Mauban, Joseph R H / Wilkinson, Katherine / Schach, Christian / Yuan, Jason X-J

American journal of physiology. Cell physiology

2005 Volume 290, Issue 2, Page(s) C325–36

Abstract: Agonist stimulation of human pulmonary artery smooth muscle cells (PASMC) and endothelial cells (PAEC) with histamine showed similar spatiotemporal patterns of Ca(2+) release. Both sustained elevation and oscillatory patterns of changes in cytosolic Ca(2+ ...

Abstract	Agonist stimulation of human pulmonary artery smooth muscle cells (PASMC) and endothelial cells (PAEC) with histamine showed similar spatiotemporal patterns of Ca(2+) release. Both sustained elevation and oscillatory patterns of changes in cytosolic Ca(2+) concentration ([Ca(2+)](cyt)) were observed in the absence of extracellular Ca(2+). Capacitative Ca(2+) entry (CCE) was induced in PASMC and PAEC by passive depletion of intracellular Ca(2+) stores with 10 microM cyclopiazonic acid (CPA; 15-30 min). The pyrazole derivative BTP2 inhibited CPA-activated Ca(2+) influx, suggesting that depletion of CPA-sensitive internal stores is sufficient to induce CCE in both PASMC and PAEC. The recourse of histamine-mediated Ca(2+) release was examined after exposure of cells to CPA, thapsigargin, caffeine, ryanodine, FCCP, or bafilomycin. In PASMC bathed in Ca(2+)-free solution, treatment with CPA almost abolished histamine-induced rises in [Ca(2+)](cyt). In PAEC bathed in Ca(2+)-free solution, however, treatment with CPA eliminated histamine-induced sustained and oscillatory rises in [Ca(2+)](cyt) but did not affect initial transient increase in [Ca(2+)](cyt). Furthermore, treatment of PAEC with a combination of CPA (or thapsigargin) and caffeine (and ryanodine), FCCP, or bafilomycin did not abolish histamine-induced transient [Ca(2+)](cyt) increases. These observations indicate that 1) depletion of CPA-sensitive stores is sufficient to cause CCE in both PASMC and PAEC; 2) induction of CCE in PAEC does not require depletion of all internal Ca(2+) stores; 3) the histamine-releasable internal stores in PASMC are mainly CPA-sensitive stores; 4) PAEC, in addition to a CPA-sensitive functional pool, contain other stores insensitive to CPA, thapsigargin, caffeine, ryanodine, FCCP, and bafilomycin; and 5) although the CPA-insensitive stores in PAEC may not contribute to CCE, they contribute to histamine-mediated Ca(2+) release.
MeSH term(s)	Adolescent ; Adult ; Animals ; Caffeine/pharmacology ; Calcium/metabolism ; Calcium Signaling/physiology ; Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone/pharmacology ; Cells, Cultured ; Endothelial Cells/cytology ; Endothelial Cells/drug effects ; Endothelial Cells/metabolism ; Endothelium, Vascular/cytology ; Endothelium, Vascular/metabolism ; Enzyme Inhibitors/pharmacology ; Female ; Histamine/metabolism ; Humans ; Indoles/pharmacology ; Ionophores/pharmacology ; Macrolides/pharmacology ; Male ; Myocytes, Smooth Muscle/cytology ; Myocytes, Smooth Muscle/drug effects ; Myocytes, Smooth Muscle/metabolism ; Pulmonary Artery/cytology ; Pulmonary Artery/metabolism ; Thapsigargin/pharmacology
Chemical Substances	Enzyme Inhibitors ; Indoles ; Ionophores ; Macrolides ; bafilomycin A (116764-51-3) ; Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone (370-86-5) ; Caffeine (3G6A5W338E) ; Thapsigargin (67526-95-8) ; Histamine (820484N8I3) ; Calcium (SY7Q814VUP) ; cyclopiazonic acid (X9TLY4580Z)
Language	English
Publishing date	2005-09-14
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	392098-7
ISSN	1522-1563 ; 0363-6143
ISSN (online)	1522-1563
ISSN	0363-6143
DOI	10.1152/ajpcell.00236.2005
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article: Non-specific binding and steric hindrance thresholds for penetration of particulate drug carriers within tumor tissue

Dancy, Jimena G / Aniket S. Wadajkar / Anthony J. Kim / Craig S. Schneider / Graeme F. Woodworth / Jeffrey A. Winkles / Joseph R.H. Mauban / Olga G. Goloubeva

Journal of Controlled Release. 2016 Sept. 28, v. 238

2016

Abstract	Therapeutic nanoparticles (NPs) approved for clinical use in solid tumor therapy provide only modest improvements in patient survival, in part due to physiological barriers that limit delivery of the particles throughout the entire tumor. Here, we explore the thresholds for NP size and surface poly(ethylene glycol) (PEG) density for penetration within tumor tissue extracellular matrix (ECM). We found that NPs as large as 62nm, but less than 110nm in diameter, diffused rapidly within a tumor ECM preparation (Matrigel) and breast tumor xenograft slices ex vivo. Studies of PEG-density revealed that increasing PEG density enhanced NP diffusion and that PEG density below a critical value led to adhesion of NP to ECM. Non-specific binding of NPs to tumor ECM components was assessed by surface plasmon resonance (SPR), which revealed excellent correlation with the particle diffusion results. Intravital microscopy of NP spread in breast tumor tissue confirmed a significant difference in tumor tissue penetration between the 62 and 110nm PEG-coated NPs, as well as between PEG-coated and uncoated NPs. SPR assays also revealed that Abraxane, an FDA-approved non-PEGylated NP formulation used for cancer therapy, binds to tumor ECM. Our results establish limitations on the size and surface PEG density parameters required to achieve uniform and broad dispersion within tumor tissue and highlight the utility of SPR as a high throughput method to screen NPs for tumor penetration.
Keywords	adhesion ; breast neoplasms ; drug carriers ; ethylene glycol ; extracellular matrix ; microscopy ; nanoparticles ; patients ; polyethylene glycol ; surface plasmon resonance
Language	English
Dates of publication	2016-0928
Size	p. 139-148.
Publishing place	Elsevier B.V.
Document type	Article
ZDB-ID	632533-6
ISSN	1873-4995 ; 0168-3659
ISSN (online)	1873-4995
ISSN	0168-3659
DOI	10.1016/j.jconrel.2016.07.034
Database	NAL-Catalogue (AGRICOLA)

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