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Intravital microscopy

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https://www.readbyqxmd.com/read/27917173/live-imaging-of-immune-responses-in-experimental-models-of-multiple-sclerosis
#1
REVIEW
Barbara Rossi, Gabriela Constantin
Experimental autoimmune encephalomyelitis (EAE) is the most common animal model of multiple sclerosis (MS), a chronic inflammatory autoimmune disease of the central nervous system (CNS) characterized by multifocal perivascular infiltrates that predominantly comprise lymphocytes and macrophages. During EAE, autoreactive T cells first become active in the secondary lymphoid organs upon contact with antigen-presenting cells (APCs), and then gain access to CNS parenchyma, through a compromised blood-brain barrier, subsequently inducing inflammation and demyelination...
2016: Frontiers in Immunology
https://www.readbyqxmd.com/read/27909339/cancer-cell-motility-lessons-from-migration-in-confined-spaces
#2
Colin D Paul, Panagiotis Mistriotis, Konstantinos Konstantopoulos
Time-lapse, deep-tissue imaging made possible by advances in intravital microscopy has demonstrated the importance of tumour cell migration through confining tracks in vivo. These tracks may either be endogenous features of tissues or be created by tumour or tumour-associated cells. Importantly, migration mechanisms through confining microenvironments are not predicted by 2D migration assays. Engineered in vitro models have been used to delineate the mechanisms of cell motility through confining spaces encountered in vivo...
December 2, 2016: Nature Reviews. Cancer
https://www.readbyqxmd.com/read/27907215/cd8-t-cells-induce-fatal-brainstem-pathology-during-cerebral-malaria-via-luminal-antigen-specific-engagement-of-brain-vasculature
#3
Phillip A Swanson, Geoffrey T Hart, Matthew V Russo, Debasis Nayak, Takele Yazew, Mirna Peña, Shahid M Khan, Chris J Janse, Susan K Pierce, Dorian B McGavern
Cerebral malaria (CM) is a severe complication of Plasmodium falciparum infection that results in thousands of deaths each year, mostly in African children. The in vivo mechanisms underlying this fatal condition are not entirely understood. Using the animal model of experimental cerebral malaria (ECM), we sought mechanistic insights into the pathogenesis of CM. Fatal disease was associated with alterations in tight junction proteins, vascular breakdown in the meninges / parenchyma, edema, and ultimately neuronal cell death in the brainstem, which is consistent with cerebral herniation as a cause of death...
December 2016: PLoS Pathogens
https://www.readbyqxmd.com/read/27904902/a-novel-hirudin-derivative-inhibiting-thrombin-without-bleeding-for-subcutaneous-injection
#4
Bing Zhao, Yanling Zhang, Yinong Huang, Jinchao Yu, Yaran Li, Qi Wang, Yixin Ma, Hou-Yan Song, Min Yu, Wei Mo
Currently, anticoagulants would be used to prevent thrombosis. Thrombin is an effector enzyme for haemostasis and thrombosis. We designed a direct thrombin inhibitor peptide (DTIP) using molecular simulation and homology modelling and demonstrated that the C-terminus of DTIP interacts with exosite I, and N-terminus with the activity site of thrombin, respectively. DTIP interfered with thrombin-mediated coagulation in human, rat and mouse plasma (n=10 per group) and blocked clotting in human whole blood in vitro...
December 1, 2016: Thrombosis and Haemostasis
https://www.readbyqxmd.com/read/27903531/dream-plays-an-important-role-in-platelet-activation-and-thrombogenesis
#5
Kyungho Kim, Alan Tseng, Andrew Barazia, Joseph E Italiano, Jaehyung Cho
Downstream regulatory element antagonist modulator (DREAM), a transcriptional repressor, is known to modulate pain responses. However, it is unknown whether DREAM is expressed in anucleate platelets and plays a role in thrombogenesis. By using intravital microscopy with DREAM-null mice and their bone marrow chimeras, we demonstrated that both hematopoietic and non-hematopoietic cell DREAM are required for platelet thrombus formation following laser-induced arteriolar injury. In a FeCl3-induced thrombosis model, we found that compared to WT control and non-hematopoietic DREAM knockout (KO) mice, DREAM KO control and hematopoietic DREAM KO mice showed a significant delay in time to occlusion...
November 30, 2016: Blood
https://www.readbyqxmd.com/read/27899358/cytohesin-1-regulates-homing-and-engraftment-of-human-hematopoietic-stem-and-progenitor-cells
#6
Justyna Rak, Katie Foster, Katarzyna Potrzebowska, Mehrnaz Safaee Talkhoncheh, Natsumi Miharada, Karolina Komorowska, Therese Torngren, Anders Kvist, Åke Borg, Lena Svensson, Dominique Bonnet, Jonas Larsson
Adhesion is a key component of hematopoietic stem cell regulation mediating homing and retention to the niche in the bone marrow. Here, using an RNA interference screen, we identify cytohesin 1 (CYTH1) as a critical mediator of adhesive properties in primary human cord-blood derived hematopoietic stem and progenitor cells (HSPCs). Knockdown of CYTH1 disrupted adhesion of HSPC to primary human mesenchymal stroma cells (MSCs). Attachment to fibronectin and ICAM1, two integrin ligands, was severely impaired, and CYTH1 deficient cells showed a reduced integrin β1 activation response, suggesting that CYTH1 mediates integrin-dependent functions...
November 29, 2016: Blood
https://www.readbyqxmd.com/read/27890922/progress-in-imaging-methods-insights-gained-into-plasmodium-biology
#7
Mariana De Niz, Paul-Christian Burda, Gesine Kaiser, Hernando A Del Portillo, Tobias Spielmann, Freddy Frischknecht, Volker T Heussler
Over the past decade, major advances in imaging techniques have enhanced our understanding of Plasmodium spp. parasites and their interplay with mammalian hosts and mosquito vectors. Cryoelectron tomography, cryo-X-ray tomography and super-resolution microscopy have shifted paradigms of sporozoite and gametocyte structure, the process of erythrocyte invasion by merozoites, and the architecture of Maurer's clefts. Intravital time-lapse imaging has been revolutionary for our understanding of pre-erythrocytic stages of rodent Plasmodium parasites...
November 28, 2016: Nature Reviews. Microbiology
https://www.readbyqxmd.com/read/27877245/imaging-nanotherapeutics-in-inflamed-vasculature-by-intravital-microscopy
#8
REVIEW
Zhenjia Wang
Intravital microscopy (IVM) is the application of light microscopy to real time study biology of live animal tissues in intact and physiological conditions with the high spatial and temporal resolution. Advances in imaging systems, genetic animal models and imaging probes, IVM has offered quantitative and dynamic insight into cell biology, immunology, neurobiology and cancer. In this review, we will focus on the targeting of nanotherapeutics to inflamed vasculature. We will introduce the basic concept and principle of IVM and demonstrate that IVM is a powerful tool used to quantitatively determine the molecular mechanisms of interactions between nanotherapeutics and neutrophils or endothelium in living mice...
2016: Theranostics
https://www.readbyqxmd.com/read/27877238/sphingomyelin-liposomes-containing-porphyrin-phospholipid-for-irinotecan-chemophototherapy
#9
Kevin A Carter, Dandan Luo, Aida Razi, Jumin Geng, Shuai Shao, Joaquin Ortega, Jonathan F Lovell
Porphyrin-phospholipid (PoP) liposomes can entrap anti-cancer agents and release them in response to near infrared (NIR) light. Doxorubicin, when remotely loaded via an ammonium sulfate gradient at a high drug-to-lipid ratio, formed elongated crystals that altered liposome morphology and could not be loaded into liposomes with higher PoP content. On the other hand, irinotecan could also be remotely loaded but did not form large crystals and did not induce liposome elongation. The loading, stability, and NIR light-triggered release of irinotecan in PoP liposomes was altered by the types of lipids used and the presence of PEGylation...
2016: Theranostics
https://www.readbyqxmd.com/read/27875293/attenuation-of-blood-brain-barrier-breakdown-and-hyperpermeability-by-calpain-inhibition
#10
Himakarnika Alluri, Marcene Grimsley, Chinchusha Anasooya Shaji, Kevin Paul Varghese, Shenyuan L Zhang, Chander Peddaboina, Bobby Robinson, Madhava R Beeram, Jason H Huang, Binu Tharakan
Blood-brain barrier (BBB) breakdown and the associated microvascular hyperpermeability followed by brain edema is a hallmark feature of several brain pathologies including traumatic brain injuries (TBI). Recent studies indicate that pro-inflammatory cytokine interleukin-1β (IL-1β) that is upregulated following traumatic injuries also promotes BBB dysfunction and hyperpermeability but the underlying mechanisms are not clearly known. The objective of this study was to determine the role of calpains in mediating BBB dysfunction and hyperpermeability and to test the effect of calpain inhibition on the BBB following traumatic insults to the brain...
November 8, 2016: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/27858361/high-resolution-intravital-microscopy-of-tumor-angiogenesis
#11
Ann L B Seynhaeve, Timo L M Ten Hagen
Real-time evaluation of vascular effects in an animal skinfold window model by intravital microscopy (IVM) provides a powerful tool to improve insight into vascular development and vascular therapy. The potential of IVM to examine processes in tissues (e.g., tumors, inflammatory sites), in a noninvasive way, enables determination of the kinetics of processes under study at any given time point. The introduction of sensitive digital cameras, confocal and multiphoton microscopy, and powerful imaging software greatly improved the quality of the images acquired...
2016: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27855336/targeting-distinct-myeloid-cell-populations-in%C3%A2-vivo-using-polymers-liposomes-and-microbubbles
#12
Can Ergen, Felix Heymann, Wa'el Al Rawashdeh, Felix Gremse, Matthias Bartneck, Ulf Panzer, Robert Pola, Michal Pechar, Gert Storm, Nicole Mohr, Matthias Barz, Rudolf Zentel, Fabian Kiessling, Christian Trautwein, Twan Lammers, Frank Tacke
Identifying intended or accidental cellular targets for drug delivery systems is highly relevant for evaluating therapeutic and toxic effects. However, limited knowledge exists on the distribution of nano- and micrometer-sized carrier systems at the cellular level in different organs. We hypothesized that clinically relevant carrier materials, differing in composition and size, are able to target distinct myeloid cell subsets that control inflammatory processes, such as macrophages, neutrophils, monocytes and dendritic cells...
November 9, 2016: Biomaterials
https://www.readbyqxmd.com/read/27833475/noninvasive-transdermal-vaccination-using-hyaluronan-nanocarriers-and-laser-adjuvant
#13
Ki Su Kim, Hyemin Kim, Yunji Park, Won Ho Kong, Seung Woo Lee, Sheldon J J Kwok, Sei Kwang Hahn, Seok Hyun Yun
Vaccines are commonly administered by injection using needles. Although transdermal microneedles are less-invasive promising alternatives, needle-free topical vaccination without involving physical damage to the natural skin barrier is still sought after as it can further reduce needle-induced anxiety and simply administration. However, this long-standing goal has been elusive since the intact skin is impermeable to most macromolecules. Here, we show an efficient, non-invasive transdermal vaccination in mice by employing two key innovations: first, the use of hyaluronan (HA) as vaccine carriers and, second, non-ablative laser adjuvants...
April 19, 2016: Advanced Functional Materials
https://www.readbyqxmd.com/read/27831589/the-anti-inflammatory-vasostatin-2-attenuates-atherosclerosis-in-apoe-mice-and-inhibits-monocyte-macrophage-recruitment
#14
Weixin Xiong, Xiaoqun Wang, Daopeng Dai, Bao Zhang, Lin Lu, Rong Tao
We showed previously that reduced level of vasostatin-2 (VS-2) correlates to the presence and severity of coronary artery disease. In this study, we aimed to figure out the role of chromogranin A (CGA) derived VS-2 in the development of atherosclerosis and monocyte/macrophage recruitment. Apolipoprotein E-deficient (ApoE(-/-)) mice fed a high-fat diet exhibited attenuated lesion size by 65 % and 41 % in En face and aortic root Oil red O staining, MOMA-2 positive area by 64 %, respectively, in VS-2 treatment group compared with PBS group...
November 10, 2016: Thrombosis and Haemostasis
https://www.readbyqxmd.com/read/27826792/monocyte-behaviour-and-tissue-transglutaminase-expression-during-experimental-autoimmune-encephalomyelitis-in-transgenic-cx3cr1-gfp-gfp-mice
#15
Navina L Chrobok, Alexandre Jaouen, Keith K Fenrich, John G J M Bol, Micha M M Wilhelmus, Benjamin Drukarch, Franck Debarbieux, Anne-Marie van Dam
Leukocyte infiltration into the central nervous system (CNS) is a key pathological feature in multiple sclerosis (MS) and the MS animal model experimental autoimmune encephalomyelitis (EAE). Recently, preventing leukocyte influx into the CNS of MS patients is the main target of MS therapies and insight into cell behaviour in the circulation is needed for further elucidation of such therapies. In this study, we aimed at in vivo visualization of monocytes in a time-dependent manner during EAE. Using intravital two-photon microscopy (IVM), we imaged CX3CR1(gfp/gfp) mice during EAE, visualizing CX3CR1-GFP(+) monocytes and their dynamics in the spinal cord vasculature...
November 9, 2016: Amino Acids
https://www.readbyqxmd.com/read/27823622/main-pathways-of-action-of-brazilian-red-propolis-on-the-modulation-of-neutrophils-migration-in-the-inflammatory-process
#16
Bruno Bueno-Silva, Marcelo Franchin, Claudiney de Freitas Alves, Carina Denny, David Fernando Colón, Thiago Mattar Cunha, Severino Matias Alencar, Marcelo Henrique Napimoga, Pedro Luiz Rosalen
BACKGROUND: Brazilian propolis is popularly used as treatment for different diseases including the ones with inflammatory origin. Brazilian red propolis chemical profile and its anti-inflammatory properties were recently described however, its mechanism of action has not been investigated yet. AIM: Elucidate Brazilian red propolis major pathways of action on the modulation of neutrophil migration during the inflammatory process. METHODS: The ethanolic extract of propolis (EEP) activity was investigated for neutrophil migration into the peritoneal cavity, intravital microscopy (rolling and adhesion of leukocytes), quantification of cytokines TNF-α, IL-1β and chemokines CXCL1/KC, CXCL2/MIP-2, neutrophil chemotaxis induced by CXCL2/MIP-2, calcium influx and CXCR2 expression on neutrophils...
December 1, 2016: Phytomedicine: International Journal of Phytotherapy and Phytopharmacology
https://www.readbyqxmd.com/read/27819526/the-effects-of-hydrogen-sulfide-on-platelet-leukocyte-aggregation-and-microvascular-thrombolysis
#17
Eberhard Grambow, Christian Leppin, Katja Leppin, Günther Kundt, Ernst Klar, Marcus Frank, Brigitte Vollmar
The volatile transmitter hydrogen sulfide (H2S) is known for its various functions in vascular biology. This study evaluates the effect of the H2S-donor GYY4137 (GYY) on thrombus stability and microvascular thrombolysis. Human whole blood served for all in vitro studies and was analyzed in a resting state, after stimulation with thrombin-receptor activating peptide (TRAP) and after incubation with 10 or 30 mM GYY or its vehicle DMSO following TRAP-activation, respectively. As a marker for thrombus stability, platelet-leukocyte aggregation was assessed using flow cytometry after staining of human whole blood against CD62P and CD45, respectively...
November 7, 2016: Platelets
https://www.readbyqxmd.com/read/27798269/transient-global-cerebral-ischemia-induces-rapid-and-sustained-reorganization-of-synaptic-structures
#18
Lirui Zhu, Lei Wang, Furong Ju, Yanli Ran, Cong Wang, Shengxiang Zhang
Ischemia can cause rapid neuronal damage. Previous studies have suggested that synaptic structures and cortical functions can be rescued if therapeutic interventions are applied in time, but the structural basis for this resilience remains incompletely understood. Here, we investigated the restoration of synaptic structures and postischemic plasticity of dendritic spines in the somatosensory cortices of mice by taking advantage of a reversible global cerebral ischemia model. Intravital two-photon imaging revealed that although dendritic structures were rapidly distorted after global ischemia, only a small percentage of spines were actually lost after transient ischemia...
October 19, 2016: Journal of Cerebral Blood Flow and Metabolism
https://www.readbyqxmd.com/read/27795303/antibody-based-immunotherapy-to-treat-and-prevent-infection-with-hypervirulent-klebsiella-pneumoniae
#19
Elizabeth Diago-Navarro, Isabel Calatayud-Baselga, Donglei Sun, Camille Khairallah, Inderjit Mann, Amaia Ulacia-Hernando, Brian Sheridan, Meiqing Shi, Bettina C Fries
Hypervirulent Klebsiella pneumoniae (hvKp) strains are predicted to become a major threat in Asia if antibiotic resistance continues to spread. Anticapsular antibodies (Abs) were developed because disseminated infections caused by hvKp are associated with significant morbidity and mortality even with antibiotic sensitive strains. K1-serotype polysaccharide capsules (K1-CPS) are expressed by the majority of hvKp strains. In this study K1-CPS specific IgG Abs were generated by conjugation of K1-CPS to immunogenic Anthrax Protective Antigen (PA) protein...
October 26, 2016: Clinical and Vaccine Immunology: CVI
https://www.readbyqxmd.com/read/27794222/neovasculature-and-circulating-tumor-cells-dual-targeting-nanoparticles-for-the-treatment-of-the-highly-invasive-breast-cancer
#20
Jianhui Yao, Jingxian Feng, Xiaoling Gao, Dan Wei, Ting Kang, Qianqian Zhu, Tianze Jiang, Xunbin Wei, Jun Chen
Antiangiogenesis therapy has been served as a potent cancer treatment strategy for decades, yet disrupting neovasculature would provoke tumor cells into invasive growth and result in distal metastasis. The basic cause of cancer metastasis can be traced down to the presence of circulating tumor cells (CTCs) which detach from primary tumor site and act as 'seeds'. Epithelial cell adhesion molecule (EpCAM) is a potential biomarker for selective capture of epithelium-derived CTCs. Here, we integrated tumor neovessles-targetable ligands K237 peptide with Ep23 aptamer against EpCAM into a single drug-loaded nanoplatform using paclitaxel (PTX) as the model drug, aiming at damaging the primary tumor and neutralizing CTCs simultaneously to achieve a synergistic anti-tumor therapeutic effect...
October 22, 2016: Biomaterials
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