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Intravital microscopy

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https://www.readbyqxmd.com/read/28336538/effects-of-dobutamine-on-intestinal-microvascular-blood-flow-heterogeneity-and-oxygen-extraction-during-septic-shock
#1
Gustavo Adolfo Ospina-Tascón, Alberto Federico García Marín, Gabriel J Echeverri, William Fernando Bermúdez, Humberto José Madriñán Navia, Juan David Valencia, Edgardo Quiñones, Fernando Rodríguez, Angela Marulanda, César Augusto Arango Davila, Alejandro Bruhn, Glenn Hernández, Daniel De Backer
Derangements of microvascular blood flow distribution might contribute to disturbing oxygen extraction by peripheral tissues. We evaluated the dynamic relationships between the mesenteric oxygen extraction ratio (mes-ERO2) and the heterogeneity of microvascular blood flow at the gut and sublingual mucosa, during the development and resuscitation of septic shock in a swine model of fecal peritonitis. Jejunal-villi and sublingual microcirculation were evaluated using a portable intravital-microscopy technique...
March 23, 2017: Journal of Applied Physiology
https://www.readbyqxmd.com/read/28330614/a-predictive-3d-multi-scale-model-of-biliary-fluid-dynamics-in-the-liver-lobule
#2
Kirstin Meyer, Oleksandr Ostrenko, Georgios Bourantas, Hernan Morales-Navarrete, Natalie Porat-Shliom, Fabian Segovia-Miranda, Hidenori Nonaka, Ali Ghaemi, Jean-Marc Verbavatz, Lutz Brusch, Ivo Sbalzarini, Yannis Kalaidzidis, Roberto Weigert, Marino Zerial
Bile, the central metabolic product of the liver, is transported by the bile canaliculi network. The impairment of bile flow in cholestatic liver diseases has urged a demand for insights into its regulation. Here, we developed a predictive 3D multi-scale model that simulates fluid dynamic properties successively from the subcellular to the tissue level. The model integrates the structure of the bile canalicular network in the mouse liver lobule, as determined by high-resolution confocal and serial block-face scanning electron microscopy, with measurements of bile transport by intravital microscopy...
March 15, 2017: Cell Systems
https://www.readbyqxmd.com/read/28329012/tracking-the-stochastic-fate-of-cells-of-the-renin-lineage-after-podocyte-depletion-using-multicolor-reporters-and-intravital-imaging
#3
Natalya V Kaverina, Hiroyuki Kadoya, Diana G Eng, Michael E Rusiniak, Maria Luisa S Sequeira-Lopez, R Ariel Gomez, Jeffrey W Pippin, Kenneth W Gross, Janos Peti-Peterdi, Stuart J Shankland
Podocyte depletion plays a major role in focal segmental glomerular sclerosis (FSGS). Because cells of the renin lineage (CoRL) serve as adult podocyte and parietal epithelial cell (PEC) progenitor candidates, we generated Ren1cCre/R26R-ConfettiTG/WT and Ren1dCre/R26R-ConfettiTG/WT mice to determine CoRL clonality during podocyte replacement. Four CoRL reporters (GFP, YFP, RFP, CFP) were restricted to cells in the juxtaglomerular compartment (JGC) at baseline. Following abrupt podocyte depletion in experimental FSGS, all four CoRL reporters were detected in a subset of glomeruli at day 28, where they co-expressed de novo four podocyte proteins (podocin, nephrin, WT-1 and p57) and two glomerular parietal epithelial cell (PEC) proteins (claudin-1, PAX8)...
2017: PloS One
https://www.readbyqxmd.com/read/28324601/two-photon-intravital-microscopy-animal-preparation-protocol-to-study-cellular-dynamics-in-pathogenesis
#4
Erinke van Grinsven, Chloé Prunier, Nienke Vrisekoop, Laila Ritsma
Two-photon intravital microscopy (2P-IVM) is an advanced imaging platform that allows the visualization of dynamic processes at subcellular resolution in vivo. Dynamic processes like cell migration, cell proliferation, cell-cell interactions, and cell signaling have an interactive character and occur in complex environments. Hence, it is of pivotal importance to study these processes in living animals, using for example 2P-IVM. 2P-IVM can be performed on a variety of tissues, from the skin of the animal to internal organs, and a variety of methods can be utilized to perform 2P-IVM on these tissues...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28302706/acclimatization-of-the-systemic-microcirculation-to-alveolar-hypoxia-is-mediated-by-inos-dependent-increased-nitric-oxide-availability
#5
Alfred J Casillan, Jie Chao, John G Wood, Norberto C Gonzalez
Rats breathing 10% O2 show a rapid and widespread systemic microvascular inflammation that results from nitric oxide (NO) depletion secondary to increased reactive O2 species (ROS) generation. The inflammation eventually resolves and the microcirculation becomes resistant to more severe hypoxia. These experiments were directed to determine the mechanisms underlying this microvascular acclimatization process. Intravital microscopy of the mesentery showed that after three weeks of hypoxia (PB ~380 Torr, inspired PO2 ~ 68-70 Torr) rats showed no evidence of inflammation; however, treatment with the inducible NO synthase (iNOS) inhibitor L-N(6)-(1-Iminoethyl) lysine dihydrochloride (L-NIL) led to ROS generation, leukocyte-endothelial adherence and emigration, and increased vascular permeability...
March 16, 2017: Journal of Applied Physiology
https://www.readbyqxmd.com/read/28302649/endothelial-protective-monocyte-patrolling-in-large-arteries-intensified-by-western-diet-and-atherosclerosis
#6
Amado Quintar, Sara McArdle, Dennis Wolf, Alex Marki, Erik Ehinger, Melanie Vassallo, Jacqueline F Miller, Zbigniew Mikulski, Klaus Ley, Konrad Buscher
Rationale: Non-classical mouse monocyte (CX3CR1(high), Ly-6C(low)) patrolling along the vessels of the microcirculation is critical for endothelial homeostasis and inflammation. Due to technical challenges it is currently not established how patrolling occurs in large arteries. Objective: This study was undertaken to elucidate the molecular, migratory and functional phenotype of patrolling monocytes in the high shear and pulsatile environment of large arteries in healthy, hyperlipidemic and atherosclerotic conditions...
March 16, 2017: Circulation Research
https://www.readbyqxmd.com/read/28300547/computer-aided-quantification-of-microvascular-networks-application-to-alterations-due-to-pathological-angiogenesis-in-the-hamster
#7
Carlos A Bulant, Pablo J Blanco, Lucas O Müller, Julio Scharfstein, Erik Svensjö
Angiogenesis is both a physiological and a pathological process of great complexity, which is difficult to measure objectively and automatically. The hamster cheek pouch (HCP) prepared for intravital-microscopy (IVM) has been used to characterize microvascular functions in many studies and was chosen to investigate microvascular characteristics observed in normal non-infected hamsters as compared to those HCPs parasitized by Trypanosoma cruzi. Images of HCPs captured at IVM were subjected to computer based measurements of angiogenesis and histamine-induced macromolecular (FITC-dextran) leakage with an image segmentation approach that has the capacity to discriminate between fluorescence emitted by macromolecular tracers inside the vasculature and in the extravascular space...
March 12, 2017: Microvascular Research
https://www.readbyqxmd.com/read/28299776/toll-like-receptors-elicit-different-recruitment-kinetics-of-monocytes-and-neutrophils-in-mouse-acute-inflammation
#8
Beat A Imhof, Stephane Jemelin, Yalin Emre
Leukocyte recruitment is an important process in combatting pathogens. The largest class of circulating leukocytes are neutrophils, which rapidly invade inflamed tissue, followed by inflammatory Ly6C(+) monocytes. Ly6C(low) monocytes patrol the endothelial wall routinely in the steady state. We recently reported early luminal recruitment of Ly6C(low) monocytes, which preceded and orchestrated neutrophil arrival and extravasation in response to TLR7/8-mediated vascular inflammation. Here we dissected the kinetics of recruitment of monocytes and neutrophils and examined the dynamics of Ly6C(low) monocytes in response to several other Toll-like receptor (TLR) agonists, using intravital confocal microscopy...
March 16, 2017: European Journal of Immunology
https://www.readbyqxmd.com/read/28296053/vasoactive-effect-of-fibronectin-derived-epiviosamine-1-and-related-peptides-in-quiescent-and-stress-models
#9
Mary D Frame, Fubao Lin, Anthony M Dewar, Richard Af Clark
OBJECTIVE: Following thermal burn injury, plasma fibronectin degrades within the interstitium; one possible product is epiviosamine-1 (EVA-1, PSHISKYILRWRPK) found within the first type III repeat of plasma fibronectin (FNIII1 ). EVA-1 ameliorates thermal burn injury progression, and binds to and enhances platelet-derived growth factor-BB (PDGF-BB) in promoting cell metabolism, growth and survival; shorter related peptides lose these abilities. Here we study the effect of EVA-1 and shorter peptides for their vasoactivity under quiescent and stress conditions...
March 15, 2017: Microcirculation: the Official Journal of the Microcirculatory Society, Inc
https://www.readbyqxmd.com/read/28287124/endogenous-tnf%C3%AE-orchestrates-the-trafficking-of-neutrophils-into-and-within-lymphatic-vessels-during-acute-inflammation
#10
Samantha Arokiasamy, Christian Zakian, Jessica Dilliway, Wen Wang, Sussan Nourshargh, Mathieu-Benoit Voisin
Neutrophils are recognised to play a pivotal role at the interface between innate and acquired immunities following their recruitment to inflamed tissues and lymphoid organs. While neutrophil trafficking through blood vessels has been extensively studied, the molecular mechanisms regulating their migration into the lymphatic system are still poorly understood. Here, we have analysed neutrophil-lymphatic vessel interactions in real time and in vivo using intravital confocal microscopy applied to inflamed cremaster muscles...
March 13, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28276477/in-vivo-longitudinal-visualization-of-bone-marrow-engraftment-process-in-mouse-calvaria-using-two-photon-microscopy
#11
Viet-Hoan Le, Seunghun Lee, Seungwon Lee, Taejun Wang, Won Hyuk Jang, Yeoreum Yoon, Soonjae Kwon, Hyekang Kim, Seung-Woo Lee, Ki Hean Kim
Intravital microscopy of mouse calvarial bone marrow (BM) is a powerful method for studying hematopoietic stem cells (HSCs) and the BM microenvironment at the cellular level. However, the current method used to access the mouse calvaria allows for only a few imaging times in the same mouse because of scar formation and inflammation induced by multiple surgeries. Longitudinal imaging of the BM may help better understand its microenvironment. In this study, a mouse calvarial window model was developed for longitudinal imaging that involves attaching a cover glass window onto the mouse calvaria and sealing the surrounding exposed area with cyanoacrylate glue and dental cement...
March 9, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28255726/in-vivo-imaging-of-t-cell-immunological-synapses-and-kinapses-in-lymph-nodes
#12
Hélène D Moreau, Philippe Bousso
T cells can become activated in lymph nodes following a diverse set of interactions with antigen-presenting cells. These cellular contacts range from short and dynamic to stable and long-lasting interactions, termed kinapses and synapses, respectively. Here, we describe a methodology to generate naïve T cells expressing a fluorescent probe of interest through the generation of bone marrow chimeras and to image T cell dynamics using intravital two-photon microscopy. In these settings, the formation of kinapses and synapses can be triggered by the administration of low and high affinity peptides, respectively...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28253383/motion-characterization-scheme-to-minimize-motion-artifacts-in-intravital-microscopy
#13
Sungon Lee, Gabriel Courties, Matthias Nahrendorf, Ralph Weissleder, Claudio Vinegoni
No abstract text is available yet for this article.
March 1, 2017: Journal of Biomedical Optics
https://www.readbyqxmd.com/read/28250053/two-photon-intravital-fluorescence-lifetime-imaging-of-the-kidney-reveals-cell-type-specific-metabolic-signatures
#14
Takashi Hato, Seth Winfree, Richard Day, Ruben M Sandoval, Bruce A Molitoris, Mervin C Yoder, Roger C Wiggins, Yi Zheng, Kenneth W Dunn, Pierre C Dagher
In the live animal, tissue autofluorescence arises from a number of biologically important metabolites, such as the reduced form of nicotinamide adenine dinucleotide. Because autofluorescence changes with metabolic state, it can be harnessed as a label-free imaging tool with which to study metabolism in vivo Here, we used the combination of intravital two-photon microscopy and frequency-domain fluorescence lifetime imaging microscopy (FLIM) to map cell-specific metabolic signatures in the kidneys of live animals...
March 1, 2017: Journal of the American Society of Nephrology: JASN
https://www.readbyqxmd.com/read/28235862/reactive-oxygen-species-producing-myeloid-cells-act-as-a-bone-marrow-niche-for-sterile-inflammation-induced-reactive-granulopoiesis
#15
Haiyan Zhu, Hyun-Jeong Kwak, Peng Liu, Besnik Bajrami, Yuanfu Xu, Shin-Young Park, Cesar Nombela-Arrieta, Subhanjan Mondal, Hiroto Kambara, Hongbo Yu, Li Chai, Leslie E Silberstein, Tao Cheng, Hongbo R Luo
Both microbial infection and sterile inflammation augment bone marrow (BM) neutrophil production, but whether the induced accelerated granulopoiesis is mediated by a common pathway and the nature of such a pathway are poorly defined. We recently established that BM myeloid cell-derived reactive oxygen species (ROS) externally regulate myeloid progenitor proliferation and differentiation in bacteria-elicited emergency granulopoiesis. In this article, we show that BM ROS levels are also elevated during sterile inflammation...
April 1, 2017: Journal of Immunology: Official Journal of the American Association of Immunologists
https://www.readbyqxmd.com/read/28223280/endothelial-cd99-supports-arrest-of-mouse-neutrophils-in-venules-and-binds-to-neutrophil-pilrs
#16
Debashree Goswami, Sigrid März, Yu-Tung Li, Annette Artz, Kerstin Schäfer, Ruth Seelige, Mariana Pacheco-Blanco, Ding Jing, Maria Gabriele Bixel, Masatake Araki, Kimi Araki, Kenichi Yamamura, Dietmar Vestweber
CD99 is a crucial regulator of the transmigration (diapedesis) of leukocytes through the blood vessel wall. Here, we report that CD99 acts at two different steps in the extravasation process. In agreement with previous antibody blocking experiments, we found that CD99 gene inactivation caused neutrophil accumulation between venular endothelial cells and the basement membrane in the inflamed cremaster. Unexpectedly, we additionally found that leukocyte attachment to the luminal surface of the venular endothelium was impaired in the absence of CD99...
February 21, 2017: Blood
https://www.readbyqxmd.com/read/28216347/systemic-application-of-carbon-monoxide-releasing-molecule-3-protects-skeletal-muscle-from-ischemia-reperfusion-injury
#17
Aurelia Bihari, Gediminas Cepinskas, Thomas L Forbes, Richard F Potter, Abdel-Rahman Lawendy
OBJECTIVE: Ischemia-reperfusion (IR) is a limb- and life-threatening complication of acute limb ischemia and musculoskeletal trauma. Carbon monoxide-releasing molecules (CORMs) have recently been shown to protect microvascular perfusion and to reduce inflammation and injury in various ischemic animal models. The purpose of this study was to examine the effects of water-soluble CORM-3 on the extent of IR-induced muscle injury. METHODS: Wistar rats were randomized into three groups: sham (no ischemia), IR + CORM-3 (10 mg/kg intraperitoneally), and IR + inactive CORM-3 (iCORM-3; 10 mg/kg intraperitoneally)...
February 16, 2017: Journal of Vascular Surgery
https://www.readbyqxmd.com/read/28198734/new-frontiers-in-intravital-microscopy-of-the-kidney
#18
Andrew M Hall, Claus D Schuh, Dominik Haenni
PURPOSE OF REVIEW: Intravital imaging with multiphoton microscopy enables the detailed study of dynamic cellular processes within functioning organs in living animals, in ways that would not otherwise be possible. It therefore represents a powerful tool in translational kidney research. In this article, we will discuss several new technical developments that have significantly increased the capabilities of kidney imaging. RECENT FINDINGS: Important contemporary advances in biomedical imaging technology include longer wavelength excitation lasers, far-red emitting fluorescent reporters, highly sensitive detectors, fluorescence lifetime measurements, adaptive optics, microendoscopes, high-throughput automated analysis algorithms and tissue clearing techniques...
February 13, 2017: Current Opinion in Nephrology and Hypertension
https://www.readbyqxmd.com/read/28187268/fibroblast-growth-factor-signaling-mediates-pulmonary-endothelial-glycocalyx-reconstitution
#19
Yimu Yang, Sarah M Haeger, Matthew A Suflita, Fuming Zhang, Kyrie L Dailey, James F Colbert, Joshay A Ford, Mario A Picon, Robert S Stearman, Lei Lin, Xinyue Liu, Xiaorui Han, Robert J Linhardt, Eric P Schmidt
RATIONALE: The endothelial glycocalyx is a heparan sulfate-rich endovascular structure critical to endothelial function. Accordingly, endothelial glycocalyx degradation during sepsis contributes to tissue edema and organ injury. OBJECTIVE: We determined the endogenous mechanisms governing pulmonary endothelial glycocalyx reconstitution, and if these reparative mechanisms are impaired during sepsis. METHODS: We performed intravital microscopy of wild-type and transgenic mice to determine the rapidity of pulmonary endothelial glycocalyx reconstitution after non-septic (heparinase-III-mediated) or septic (cecal ligation and puncture-mediated) endothelial glycocalyx degradation...
February 10, 2017: American Journal of Respiratory Cell and Molecular Biology
https://www.readbyqxmd.com/read/28182324/clumping-factor-a-von-willebrand-factor-binding-protein-and-von-willebrand-factor-anchor-staphylococcus-aureus-to-the-vessel-wall
#20
J Claes, L Liesenborghs, M Peetermans, T R Veloso, D Missiakas, O Schneewind, S Mancini, J M Entenza, M F Hoylaerts, R Heying, P Verhamme, T Vanassche
OBJECTIVE: When establishing endovascular infections, Staphylococcus aureus (S. aureus) overcomes shear forces of flowing blood by binding to von Willebrand factor (VWF). Staphylococcal VWF-binding protein (vWbp) interacts with VWF, but it is unknown how this secreted protein binds to the bacterial cell wall. We hypothesized that vWbp interacts with a staphylococcal surface protein, mediating the adhesion of S. aureus to VWF and vascular endothelium under shear stress. METHODS: We studied the binding of S...
February 9, 2017: Journal of Thrombosis and Haemostasis: JTH
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