Amnis Publications

At Sigma-Aldrich, we aim to bring innovative solutions closer to you. The publications below show how our products play a part in research.

 October 2017

 

Adenomatous Polyposis Coli Defines Treg Differentiation and Anti-inflammatory Function through Microtubule-Mediated NFAT Localization
Original Research Article
Cell Reports, Volume 21, Issue 1, 3 October 2017, Pages 181-194

Sonia Agüera-González, Oliver T. Burton, Elena Vázquez-Chávez, Céline Cuche, Floriane Herit, Jérôme Bouchet, Rémi Lasserre, Iratxe del Río-Iñiguez, Vincenzo Di Bartolo, Andrés Alcover

Graphical Abstract

GalNAc bio-functionalization of nanoparticles assembled by electrostatic interactions improves siRNA targeting to the liver
Original Research Article
Journal of Controlled Release, Available online 4 October 2017, Pages

Efrat Korin, Tzlil Bejerano, Smadar Cohen

Graphical abstract

 

 September 2017

 

Are Amniotic Fluid Neutrophils in Women with Intra-Amniotic Infection and/or Inflammation of Fetal or Maternal Origin?
Original Research Article
American Journal of Obstetrics and Gynecology, Available online 28 September 2017, Pages
Nardhy Gomez-Lopez, Roberto Romero, Yi Xu, Yaozhu Leng, Valeria Garcia-Flores, Derek Miller, Suzanne M. Jacques, Sonia S. Hassan, Jonathan Faro, Adham Alsamsam, Ali Alhousseini, Hunter Gomez-Roberts, Bogdan Panaitescu, Lami Yeo, Eli Maymon
Pro-inflammatory chitosan/poly(γ-glutamic acid) nanoparticles modulate human antigen-presenting cells phenotype and revert their pro-invasive capacity
Original Research Article
Acta Biomaterialia, Available online 14 September 2017, Pages

Flávia Castro, Marta L. Pinto, Andreia M. Silva, Catarina L. Pereira, Graciosa Q. Teixeira, Maria Gomez-Lazaro, Susana G. Santos, Mário A. Barbosa, Raquel M. Gonçalves, Maria J. Oliveira

Graphical abstract

 

Myeloid leukemia factor functions in anti-WSSV immune reaction of kuruma shrimp, Marsupenaeus japonicus
Original Research Article
Fish & Shellfish Immunology, Available online 12 September 2017, Pages

Xiao-Wu Feng, Li-Jie Huo, Jie-Jie Sun, Ji-Dong Xu, Guo-Juan Niu, Jin-Xing Wang, Xiu-Zhen Shi
Indoleamine 2,3-dioxygenase 1, Increased in Human Gastric Pre-Neoplasia, Promotes Inflammation and Metaplasia in Mice and is Associated with Type II Hypersensitivity/Autoimmunity
Original Research Article
Gastroenterology, Available online 11 September 2017, Pages

Mohamad El-Zaatari, Adam J. Bass, Reanne Bowlby, Min Zhang, Li-Jyun Syu, Yitian Yang, Helmut Grasberger, Andrew Shreiner, Bei Tan, Shrinivas Bishu, Wai K. Leung, Andrea Todisco, Nobuhiko Kamada, Marilia Cascalho, Andrzej A. Dlugosz, John Y. Kao
Chromatin organization as an indicator of glucocorticoid induced natural killer cell dysfunction
Original Research Article
Brain, Behavior, and Immunity, Available online 12 September 2017, Pages

Michael S. Misale, Linda Witek Janusek, Dina Tell, Herbert L. Mathews
Multivalent interactions between streptavidin-based pretargeting fusion proteins and cell receptors impede efficient internalization of biotinylated nanoparticles
Original Research Article
Acta Biomaterialia, Available online 9 September 2017, Pages

Christina L. Parker, Qi Yang, Bing Yang, Justin D. McCallen, Steven I. Park, Samuel K. Lai

Graphical abstract

 

Distinct roles for the deacetylase domain of HDAC3 in the hippocampus and medial prefrontal cortex in the formation and extinction of memory
Original Research Article
Neurobiology of Learning and Memory, Available online 8 September 2017, Pages

Yasaman Alaghband, Janine L. Kwapis, Alberto J. López, André O. White, Osasumwen V. Aimiuwu, Amni Al-Kachak, Kasuni K. Bodinayake, Nicole C. Oparaugo, Richard Dang, Mariam Astarabadi, Dina P. Matheos, Marcelo A. Wood
Therapy with eculizumab for patients with primary p.Cys89Tyr mutation in the CD59 gene avoids recurrent arterial thrombosis
Molecular Immunology, Volume 89, September 2017, Pages 186
Dror Mevorach, Adi Tabib, Netanel Karbian
The effect of complement inhibition on erythrocyte destruction in AIHA
Molecular Immunology, Volume 89, September 2017, Pages 203
Inge Baas, Edimara S. Reis, Daniel Ricklin, John D. Lambris, Masja de Haas, Diana Wouters, Sacha S. Zeerleder
Development of a 3D model to test the effects of the endocrine disrupting chemical bisphenol A (BPA) in human placental cells
Placenta, Volume 57, September 2017, Pages 299
Sophie-Christine Jahn, Elisabete Silva, Emmanouil Karteris
Characterising the effect of antimalarial drugs on the maturation and clearance of murine blood-stage Plasmodium parasites in vivo
Original Research Article
International Journal for Parasitology, Available online 31 August 2017, Pages

David S. Khoury, Deborah Cromer, Trish Elliott, Megan S.F. Soon, Bryce S. Thomas, Kylie R. James, Shannon E. Best, Rosemary A. Aogo, Jessica A. Engel, Kate H. Gartlan, Jasmin Akter, Ismail Sebina, Ashraful Haque, Miles P. Davenport

Graphical abstract

 

Salt and cadmium stress tolerance caused by overexpression of the Glycine Max Na+/H+ Antiporter (GmNHX1) gene in duckweed (Lemna turionifera 5511)
Original Research Article
Aquatic Toxicology, Available online 26 August 2017, Pages

Lin Yang, Yujie Han, Di Wu, Wang Yong, Miaomiao Liu, Sutong Wang, Wenxin Liu, Meiyi Lu, Ying Wei, Jinsheng Sun

 

 August 2017

 

Mitochondria are implicated in the regulation of terminal erythropoiesis   
Experimental Hematology, Volume 53, Supplement, September 2017, Pages S83
Raymond Liang, Saghi Ghaffari
Regulatory Innate Lymphoid Cells Control Innate Intestinal Inflammation   
Original Research Article
Cell, Available online 24 August 2017, Pages
Shuo Wang, Pengyan Xia, Yi Chen, Yuan Qu, Zhen Xiong, Buqing Ye, Ying Du, Yong Tian, Zhinan Yin, Zhiheng Xu, Zusen Fan

Graphical Abstract

Bioactive carbon dots lights up microtubules and destabilises cell cytoskeletal framework – A robust imaging agent with therapeutic activity   
Original Research Article
Colloids and Surfaces B: Biointerfaces, Available online 19 August 2017, Pages
S. Uday Kumar, Bharat Bhushan, P. Gopinath

Graphical abstract

In vitro polyphenol effects on apoptosis: An update of literature data   
Review Article
Seminars in Cancer Biology, Available online 19 August 2017, Pages
Valeria Curti, Arianna Di Lorenzo, Marco Dacrema, Jianbo Xiao, Sayed Mohammad Nabavi, Maria Daglia
Trisomy 12 assessment by conventional fluorescence in-situ hybridization (FISH), FISH in suspension (FISH-is) and laser scanning cytometry (LSC) in chronic lymphocytic leukemia.   
Original Research Article
Cancer Genetics, Available online 4 August 2017, Pages
Cuc H. Do, Karen M. Lower, Cindy Macardle, Bryone J. Kuss
Formulation optimization, characterization, and evaluation of in vitro cytotoxic potential of Curcumin loaded solid lipid nanoparticles for improved anticancer activity   
Original Research Article
Chemistry and Physics of Lipids, Available online 23 August 2017, Pages
Sri Vishnu Kiran, Bhatt Himanshu, Aashma Shah, Neeraja Komanduri, Dhanya Vijayasarathy, Balaram Ghosh, Swati Biswas

Graphical abstract


 

 July 2017

 

Mass Cytometric Analysis of HIV Entry, Replication, and Remodeling in Tissue CD4+ T Cells   
Original Research Article
Cell Reports, Volume 20, Issue 4, 25 July 2017, Pages 984-998
Marielle Cavrois, Trambak Banerjee, Gourab Mukherjee, Nandhini Raman, Rajaa Hussien, Brandon Aguilar Rodriguez, Joshua Vasquez, Matthew H. Spitzer, Nicole H. Lazarus, Jennifer J. Jones, Christina Ochsenbauer, Joseph M. McCune, Eugene C. Butcher, Ann M. Arvin, Nandini Sen, Warner C. Greene, Nadia R. Roan

Graphical Abstract

An Efficient Platform for Astrocyte Differentiation from Human Induced Pluripotent Stem Cells   
Original Research Article
Stem Cell Reports, Available online 27 July 2017, Pages
Julia TCW, Minghui Wang, Anna A. Pimenova, Kathryn R. Bowles, Brigham J. Hartley, Emre Lacin, Saima I. Machlovi, Rawan Abdelaal, Celeste M. Karch, Hemali Phatnani, Paul A. Slesinger, Bin Zhang, Alison M. Goate, Kristen J. Brennand
The Tim-3-galectin-9 Secretory Pathway is Involved in the Immune Escape of Human Acute Myeloid Leukemia Cells   
Original Research Article
EBioMedicine, Available online 19 July 2017, Pages
Isabel Gonçalves Silva, Inna M. Yasinska, Svetlana S. Sakhnevych, Walter Fiedler, Jasmin Wellbrock, Marco Bardelli, Luca Varani, Rohanah Hussain, Giuliano Siligardi, Giacomo Ceccone, Steffen M. Berger, Yuri A. Ushkaryov, Bernhard F. Gibbs, Elizaveta Fasler-Kan, Vadim V. Sumbayev
Peroxisome-Mediated Metabolism Is Required for Immune Response to Microbial Infection   
Original Research Article
Immunity, Volume 47, Issue 1, 18 July 2017, Pages 93-106.e7
Francesca Di Cara, Avinash Sheshachalam, Nancy E. Braverman, Richard A. Rachubinski, Andrew J. Simmonds

Graphical Abstract

Discrete β-adrenergic mechanisms regulate early and late erythropoiesis in erythropoietin-resistant anemia   
Original Research Article
Surgery, Available online 14 July 2017, Pages
Shirin Hasan, Michael J. Mosier, Andrea Szilagyi, Richard L. Gamelli, Kuzhali Muthumalaiappan
Myeloid-derived suppressor cells modulate B-cell responses   
Original Research Article
Immunology Letters, Available online 4 July 2017, Pages
Felipe J.N. Lelis, Jennifer Jaufmann, Anurag Singh, Katja Fromm, Annkathrin Chiara Teschner, Simone Pöschel, Iris Schäfer, Sandra Beer-Hammer, Nikolaus Rieber, Dominik Hartl

 

 May 2017

 

Effects of cyanobacteria Oscillatoria sp. lipopolysaccharide on B cell activation and Toll-like receptor 4 signaling   
Original Research Article
Toxicology Letters, Volume 275, 5 June 2017, Pages 101-107
Michelle Swanson-Mungerson, Ryan Incrocci, Vijay Subramaniam, Philip Williams, Mary L. Hall, Alejandro M.S. Mayer
Nuclei Size Distribution as a Predictor for Radiosensitivity with 192Ir Brachytherapy   
Brachytherapy, Volume 16, Issue 3, Supplement, May–June 2017, Pages S83
Nguyen Truong, Michael Evans, Norma Ybarra, Shirin Enger
The Impact of Autophagy on the Progression of the Myelodysplastic Syndromes   
Leukemia Research, Volume 55, Supplement 1, April 2017, Pages S127
F.S. Vilcassim, A. Banerjee, G. Grigoriadis
Structure-activity relationship study of the tumour-targeting peptide A20FMDV2 via modification of Lys16, Leu13, and N- and/or C-terminal functionality   
Original Research Article
European Journal of Medicinal Chemistry, Available online 3 May 2017, Pages
Kuo-yuan Hung, Paul W.R. Harris, Ami Desai, John F. Marshall, Margaret A. Brimble
Characterization of Protein Particles in Therapeutic Formulations using Imaging Flow Cytometry   
Original Research Article
Journal of Pharmaceutical Sciences, Available online 27 April 2017, Pages
Christine Probst, Yuanchun Zeng, Rong-Rong Zhu
Contribution of leukocytes to the induction and resolution of the acute inflammatory response in chickens   
Original Research Article
Developmental & Comparative Immunology, Available online 26 April 2017, Pages
Juan A. More Bayona, Anbu Kumar Karuppannan, Daniel R. Barreda
Current approaches for avoiding the limitations of CTC detection methods; implications for diagnosis and treatment of patients with solid tumors.   
Review Article
Translational Research, Available online 26 April 2017, Pages
Artur Kowalik, Magdalena Kowalewska, Stanisław Góźdź
Amniotic membrane transplants in the pediatric population   
Original Research Article
Journal of American Association for Pediatric Ophthalmology and Strabismus, Available online 24 April 2017, Pages
Mariam S. Ahmad, Garett S. Frank, Eric M. Hink, Alan G. Palestine, Darren G. Gregory, Emily A. McCourt

 

 April 2017

 

iPSC-Derived Human Microglia-like Cells to Study Neurological Diseases   
Original Research Article
Neuron, Volume 94, Issue 2, 19 April 2017, Pages 278-293.e9
Edsel M. Abud, Ricardo N. Ramirez, Eric S. Martinez, Luke M. Healy, Cecilia H.H. Nguyen, Sean A. Newman, Andriy V. Yeromin, Vanessa M. Scarfone, Samuel E. Marsh, Cristhian Fimbres, Chad A. Caraway, Gianna M. Fote, Abdullah M. Madany, Anshu Agrawal, Rakez Kayed, Karen H. Gylys, Michael D. Cahalan, Brian J. Cummings, Jack P. Antel, Ali Mortazavi, Monica J. Carson, Wayne W. Poon, Mathew Blurton-Jones
Examining DNA Double-Strand Break Repair in a Cell Cycle-Dependent Manner   
Methods in Enzymology, Available online 17 April 2017, Pages
Janapriya Saha, Shih-Ya Wang, Anthony J. Davis
Evaluating Marinichlorella kaistiae KAS603 cell size variation, growth and TAG accumulation resulting from rapid adaptation to highly diverse trophic and salinity cultivation regimes   
Original Research Article
Algal Research, Volume 25, July 2017, Pages 12-24
Eva L. Sánchez-Alvarez, Grisele González-Ledezma, José A. Bolaños Prats, José L. Stephano-Hornedo, Mark Hildebrand
Virgin Beta Cells Persist throughout Life at a Neogenic Niche within Pancreatic Islets   
Original Research Article
Cell Metabolism, Volume 25, Issue 4, 4 April 2017, Pages 911-926.e6
Talitha van der Meulen, Alex M. Mawla, Michael R. DiGruccio, Michael W. Adams, Vera Nies, Sophie Dólleman, Siming Liu, Amanda M. Ackermann, Elena Cáceres, Anna E. Hunter, Klaus H. Kaestner, Cynthia J. Donaldson, Mark O. Huising

Graphical Abstract

Evaluating Marinichlorella kaistiae KAS603 cell size variation, growth and TAG accumulation resulting from rapid adaptation to highly diverse trophic and salinity cultivation regimes
Original Research Article
Algal Research, Volume 25, July 2017, Pages 12-24
Eva L. Sánchez-Alvarez, Grisele González-Ledezma, José A. Bolaños Prats, José L. Stephano-Hornedo, Mark Hildebrand
Virgin Beta Cells Persist throughout Life at a Neogenic Niche within Pancreatic Islets
Original Research Article
Cell Metabolism, Volume 25, Issue 4, 4 April 2017, Pages 911-926.e6
Talitha van der Meulen, Alex M. Mawla, Michael R. DiGruccio, Michael W. Adams, Vera Nies, Sophie Dólleman, Siming Liu, Amanda M. Ackermann, Elena Cáceres, Anna E. Hunter, Klaus H. Kaestner, Cynthia J. Donaldson, Mark O. Huising

Graphical Abstract

 

 

 March 2017

 

Anti-HER2 immunoliposomes for co-delivery of paclitaxel and rapamycin for breast cancer therapy   
Original Research Article
European Journal of Pharmaceutics and Biopharmaceutics, Available online 28 February 2017, Pages
Josimar O. Eloy, Raquel Petrilli, Deise L. Chesca, Fabiano P. Saggioro, Robert J. Lee, Juliana Maldonado Marchetti
Graphical abstract

 


 

Toxic effects of perfluorinated compounds at human cellular level and on a model vertebrate   
Original Research Article
Food and Chemical Toxicology, Available online 9 March 2017, Pages
Sandra Rainieri, Nadia Conlledo, Tomaž Langerholc, Eneko Madorran, Martin Sala, Alejandro Barranco
A novel method for detection of IFN-lambda 3 binding to cells for quantifying IFN-lambda receptor expression   
Original Research Article
Journal of Immunological Methods, Available online 6 March 2017, Pages
Deanna M. Santer, Gillian E.S. Minty, Adil Mohamed, Lesley Baldwin, Rakesh Bhat, Michael Joyce, Adrian Egli, D. Lorne J. Tyrrell, Michael Houghton
Interactions between tafenoquine and artemisinin-combination therapy partner drug in asexual and sexual stage Plasmodium falciparum   
Original Research Article
International Journal for Parasitology: Drugs and Drug Resistance, Volume 7, Issue 2, August 2017, Pages 131-137
Karen Kemirembe, Mynthia Cabrera, Liwang Cui

 

Graphical abstract

 

Emerging Role of Chemoprotective Agents in the Dynamic Shaping of Plasma Membrane Organization   
Review Article
Biochimica et Biophysica Acta (BBA) - Biomembranes, Available online 22 March 2017, Pages
Natividad R. Fuentes, Michael L. Salinas, Eunjoo Kim, Robert S. Chapkin
Effects of macromolecular crowding on alkaline phosphatase unfolding, conformation and stability   
Original Research Article
International Journal of Biological Macromolecules, Available online 23 March 2017, Pages
Jiajia Jia, Xin Peng, Wei Qi, Rongxin Su, Zhimin He

 

Graphical abstract

 

 

 February 2017

 

Effects of chrysolaminarin synthase knockdown in the diatom Thalassiosira pseudonana: Implications of reduced carbohydrate storage relative to green algae
Original Research Article
Algal Research, Volume 23, April 2017, Pages 66-77

Mark Hildebrand, Kalpana Manandhar-Shrestha, Raffaela Abbriano
Neutrophils exert protection in early Aeromonas veronii infections through the clearance of both bacteria and dying macrophages
Original Research Article
Fish & Shellfish Immunology, Available online 3 February 2017, Pages

Jeffrey J. Havixbeck, Aja M. Rieger, Lucas J. Churchill, Daniel R. Barreda
Identification of high independent prognostic value of nanotechnology based circulating tumor cell enumeration in first-line chemotherapy for metastatic breast cancer patients
Original Research Article
The Breast, Volume 32, April 2017, Pages 119-125

Xiao-ran Liu, Bin Shao, Jia-xi Peng, Hui-ping Li, Yan-lian Yang, Wei-yao Kong, Guo-hong Song, Han-fang Jiang, Xu Liang, Ying Yan
Gene delivery of apoptin-derived peptide using an adeno-associated virus vector inhibits glioma and prolongs animal survival
Biochemical and Biophysical Research Communications, Volume 482, Issue 3, 15 January 2017, Pages 506-513
Xiuli Zhong, Hengyu Zhao, Songhe Liang, DanYang Zhou, Wenjia Zhang, Lijie Yuan

 

 January 2017

 

What you see matters: Enhanced detection using image-based flow cytometry

Brian K. McFarlin
 
Flow cytometry what you see matters: Enhanced clinical detection using image-based flow cytometry

Pages 1-8

Brian K. McFarlin, Melody A. Gary
• This review summarizes work published previously by all authors in this special issue.
• This review is designed to highlight the novelty of the field of image-based flow cytometry.
• This review should serve as the foundation for expanded knowledge base in the future.
Applications of imaging flow cytometry in the diagnostic assessment of acute leukaemia

Pages 39-45


Lizz F. Grimwade, Kathryn A. Fuller, Wendy N. Erber
• Imaging flow cytometry can be applied to the diagnosis of acute leukaemia.
• This can be used to detect disrupted PML bodies in acute promyelocytic leukaemia.
• Nucleophosmin localisation can be determined in acute myeloid leukaemia.
• Fluorescence in situ hybridisation can be performed on intact phenotyped cells.
• Potential to integrate all leukaemia diagnostic tests by imaging flow cytometry.
Imaging flow cytometry in the assessment of leukocyte-platelet aggregates

Pages 46-54


Henry Hui, Kathy A. Fuller, Wendy N. Erber, Matthew D. Linden



Delineating stages of erythropoiesis using imaging flow cytometry

Pages 68-74


K.E. McGrath, S.C. Catherman, J. Palis



Quantifying autophagy: Measuring LC3 puncta and autolysosome formation in cells using multispectral imaging flow cytometry

Pages 147-156


Haley R. Pugsley (Scientist from Amnis – Seattle)
• A review of the predominant methods to measure autophagy via MIFC.
• Bright Detail Intensity and Spot Count work for measuring LC3 puncta accumulation.
• Bright Detail Similarity (BDS) can measure the formation of the autolysosome.
• Spot Count vs BDS provides the most comprehensive information on autophagy.
• There is not a single correct method for measuring autophagy using MIFC.
Using image-based flow cytometry to monitor satellite cells proliferation and differentiation in vitro

Pages 175-181


Hui-Ying Luk, Brian K. McFarlin, Jakob L. Vingren

 

Consumption of a high-fat, high-calorie meal is associated with an increase in intracellular co-localization of PPAR-γ mRNA and protein in monocytes

Pages 182-187


Andrea L. Henning, Brian K. McFarlin



Imaging flow cytometry for the screening of compounds that disrupt the Plasmodium falciparum digestive vacuole

Pages 211-220


Wan Ni Chia, Yan Quan Lee, Kevin Shyong-Wei Tan
• Plasmodium parasites undergo cell death upon treatment with DV-disrupting drugs.
• Disrupted parasite DVs can be observed using a fluorescent Ca2+-binding reporter.
• This paper describes a phenotypic screen using an imaging flow cytometer (IFC).
• Autosampler fitted IFC allows high-throughput screening of big compound libraries.

 

 November 2016

 

Imaging flow cytometry and GST pulldown assays provide new insights into channel catfish leukocyte immune-type receptor-mediated phagocytic pathways

Developmental & Comparative Immunology

Myron A. Zwozdesky, Chenjie Fei, Dustin M.E. Lillico, James L. Stafford

Channel catfish (Ictalurus punctatus) leukocyte immune-type receptors (IpLITRs) control various innate immune cell effector responses including the phagocytic process. This large immunoregulatory receptor family also consists of multiple receptor-types with variable signaling abilities that is dependent on their inherent or acquired tyrosine-containing cytoplasmic tail (CYT) regions. For example, IpLITR 2.6b associates with the immunoreceptor tyrosine-based activation motif (ITAM)-containing adaptor molecule IpFcRγ-L, and when expressed in mammalian cells it activates phagocytosis using a similar profile of intracellular signaling mediators that also regulate the prototypical mammalian Fc receptor (FcR) phagocytic pathway. Alternatively, IpLITR 1.1b contains a long tyrosine-containing CYT with multifunctional capabilities including both inhibitory and stimulatory actions. Recently, we demonstrated that IpLITR 1.1b activates a unique phagocytic pathway involving the generation of multiple plasma membrane extensions that rapidly capture extracellular targets and secure them on the cell surface in phagocytic cup-like structures. Occasionally, these captured targets are completely engulfed albeit at a significantly lower rate than what was observed for IpLITR 2.6b. While this novel IpLITR 1.1b phagocytic activity is insensitive to classical blockers of phagocytosis, its distinct target capture and engulfment actions depend on the engagement of the actin polymerization machinery. However, it is not known how this protein translates target recognition into intracellular signaling events during this atypical mode of phagocytosis. Using imaging flow cytometry and GST pulldown assays, the aims of this study were to specifically examine what regions of the IpLITR 1.1b CYT trigger phagocytosis and to establish what profile of intracellular signaling molecules likely participate in its actions. Our results show that in stably transfected AD293 cells, the membrane proximal and distal CYT segments of IpLITR 1.1b independently regulate its phagocytic activities. These CYT regions were also shown to differentially recruit various SH2 domain-containing intracellular mediators, which provides new information about the dynamic immunoregulatory abilities of IpLITR 1.1b. Overall, this work further advances our understanding of how certain immunoregulatory receptor-types link extracellular target binding events to the actin polymerization machinery during a non-classical mode of phagocytosis.

Pyruvate dehydrogenase has a major role in mast cell function and its activity is regulated by mitochondrial MITF

Journal of Allergy and Clinical Immunology

Israa Sharkia, Tal Hadad Erlich, Nadine Landolina, Miri Assayag, Alex Motzik, Inbal Rachmin, Gillian Kay, Ziv Porat, Sagi Tshori, Neville Berkman, Francesca Levi-Schaffer, Ehud Razin

We have recently observed that OXPHOS mediated ATP production is essential for mast cell function. Pyruvate dehydrogenase (PDH) is the main regulator of the Krebs cycle and is located upstream to the electron transport chain. However, the role of PDH in mast cell function has not been described. Microphthalmia transcription factor (MITF) regulates the development, number and function of mast cells. The localization of MITF to the mitochondria and its interaction with mitochondrial proteins has not been explored.

To explore the role played by PDH in mast cell exocytosis and to determine whether MITF is localized in the mitochondria and is involved in the regulation of PDH activity.

Experiments were performed in vitro using human and mouse mast cells as well as RBL cells, and in vivo in mice. The effect of PDH inhibition on mast cell function was examined. PDH interaction with MITF was measured before and after immunological activation. Furthermore, mitochondrial localization of MITF and its effect on PDH activity were determined.

PDH is essential for immunologically mediated degranulation of mast cells. Following activation PDH is serine dephosphorylated. In addition, we show for the first time that MITF is partially located in the mitochondria and interacts with PDH. This interaction is dependent on the phosphorylation state of PDH. Furthermore, mitochondrial MITF regulates PDH activity.

The association of mitochondrial MITF with PDH emerges as an important regulator of mast cell function. Our findings indicate that PDH could arise as a new target for the manipulation of allergic diseases.

 

 October 2016

 

Title Abstract
Mechanisms of cellular uptake and endosomal escape of calcium-siRNA nanocomplexes

International Journal of Pharmaceutics, Volume 515, Issues 1–2, 30 December 2016, Pages 46-56

Matan Goldshtein (a), Efrat Forti (a), Emil Ruvinov (a), Smadar Cohen (a) (b) (c)
a Avram and Stella Goldstein-Goren Department of Biotechnology Engineering, Ben-Gurion University of the Negev, Beer-Sheva, Israel
b Regenerative Medicine and Stem Cell (RMSC) Research Center, Ben-Gurion University of the Negev, Beer-Sheva, Israel
c The Ilse Katz Institute for Nanoscale Science and Technology, Ben-Gurion University of the Negev, Beer-Sheva, Israel
Ca2+-siRNA nanocomplexes represent a simple yet an effective platform for siRNA delivery into the cell cytoplasm, with subsequent successful siRNA-induced target gene silencing. Herein, we aimed to elucidate the roles played by calcium ions in siRNA nanocomplex formation, cell uptake, and endosomal escape. We investigated whether the replacement of Ca2+in the nanocomplex by other bivalent cations would affect their cell entry and subsequent gene silencing. Our results indicate that Mg2+ and Ba2+ lead to the formation of nanocomplexes of similar physical features (size = 100 nm, surface charge ζ = −8 mV) as the Ca2+-siRNA nanocomplexes. Yet, these nanocomplexes were not uptaken by the cells to the same extent as those prepared with Ca2+, and siRNA-induced target gene silencing was not obtained. Cell internalization of Ca2+−-siRNA nanocomplexes, examined by employing chemical inhibitors to clathrin-, caveolin- and dynamin-mediated endocytosis pathways, indicated the involvement of all mechanisms in the process. Inhibition of endosome acidification by bafilomycin completely abolished the siRNA-mediated silencing by Ca2+-siRNA nanocomplexes. Collectively, our results indicate that Ca2+ promotes cell internalization and rapid endosomal escape, thus leading to the efficient siRNA-induced target gene silencing elicited by the Ca2+-siRNA nanocomplexes.
Vibration and buckling analysis of partially cracked thin orthotropic rectangular plates in thermal environment

Thin-Walled Structures, Volume 109, December 2016, Pages 143-158

P.V. Joshi (a), N.K. Jain (b), G.D. Ramtekkar (c), Gurveer Singh Virdi (a)
a Department of Mechanical Engineering, SSTC,SSGI, Bhilai, C.G., India
b Department of Mechanical Engineering, National Institute of Technology, Raipur, C.G., India
c Department of Civil Engineering, National Institute of Technology, Raipur, C.G., India
An analytical model is presented for vibration of a thin orthotropic plate containing two perpendicular continuous line surface cracks located at the centre of the plate in the presence of thermal environment. Also new configuration of two perpendicular cracks as internal cracks, located along the thickness of the plate is studied by considering appropriate crack compliance coefficients based on line-spring model. Equilibrium principle based on Classical Plate Theory is used to derive the equations of motion for cracked plate, wherein the crack terms are formulated using the line spring model. The moments due to thermal environment are neglected in the results and only uniform heating of the cracked plate is considered. The solution for natural frequencies of cracked plate is obtained by Galerkin’s method. A relation for thermal buckling phenomenon for the cracked plate is also formulated. The influence of the lengths of the two cracks and their location along the thickness of the plate on critical buckling temperature and first mode natural frequency is demonstrated. The geometrically linear frequency response relation for cracked plate is formulated using the method of multiple scales. It is thus concluded that the presence of the two cracks affect the critical buckling temperature and natural frequencies.
Imaging flow cytometry for the characterization of extracellular vesicles

Methods, Available online 6 October 2016

Joanne Lannigan (a), Uta Erdbruegger (b)
a University of Virginia, School of Medicine, Flow Cytometry Core, 1300 Jefferson Park Avenue, Charlottesville, VA 22908-0734, USA
b University of Virginia, Department of Medicine/Nephrology Division, 1300 Jefferson Park Avenue, Charlottesville, VA 22908-0133, USA
Extracellular Vesicles (EVs) are potent bio-activators and inter-cellular communicators that play an important role in both health and disease. It is for this reason there is a strong interest in understanding their composition and origin, with the hope of using them as important biomarkers or therapeutics. Due to their very small size, heterogeneity, and large numbers there has been a need for better tools to measure them in an accurate and high throughput manner. While traditional flow cytometry has been widely used for this purpose, there are inherent problems with this approach, as these instruments have traditionally been developed to measure whole cells, which are orders of magnitude larger and express many more molecules of identifying epitopes.

Imaging flow cytometry, as performed with the ImagestreamX MKII, with its combination of increased fluorescence sensitivity, low background, image confirmation ability and powerful data analysis tools, provides a great tool to accurately evaluate EVs. We present here a comprehensive approach in applying this technology to the study of EVs.
Imaging flow cytometry in the assessment of leukocyte-platelet aggregates

Methods, Available online 5 October 2016

Henry Hui, Kathy A. Fuller, Wendy N. Erber, Matthew D. Linden,
The University of Western Australia, Australia
Platelets are subcellular blood elements with a well-established role in haemostasis. Upon activation platelets undergo granule exocytosis, resulting in α-granule P-Selectin being expressed on the cell membrane. This allows binding of activated platelets to P-Selectin glycoprotein ligand 1 (PSGL-1) expressing leukocytes, forming leukocyte-platelet aggregates (LPAs). Whole blood flow cytometry (FCM) has demonstrated that elevated circulating LPAs (especially monocyte LPAs) are linked to atherothrombosis in high risk patients, and that activated platelet binding influences monocytes towards a pro-adhesive and pro-atherogenic phenotype. However, a limitation of conventional FCM is the potential for coincident events to resemble LPAs despite no tethering. Imaging cytometry can be used to characterize LPA formation and distinguish circulating MPAs from coincidental events. Platelets and leukocyte subsets are identified by expression of surface markers (e.g. the lipopolysachharide receptor CD14 on monocytes, glycoprotein Ib CD42b on platelets). In conventional FCM, all events with both leukocyte and platelet characteristics are designated as LPAs. However, by using an ‘internal’ mask based on the brightfield image and the fluorescent platelet identifier, imaging flow cytometry is able to distinguish leukocytes with tethered platelets (genuine LPAs) from leukocyte with coincidental, untethered platelets nearby. Mechanisms (e.g. adhesion molecules) or consequences (e.g. signal transduction) can then be separately analysed in platelet tethered and untethered leukocytes. Imaging flow cytometry therefore provides a more accurate approach for both enumeration and analysis of LPAs than conventional FCM.
Insights into the interaction of CD4 with anti-CD4 antibodies

Immunobiology, Available online 17 October 2016, Pages

Van-Chien Bui (a) (b), Thi-Huong Nguyen (b)
a Center for Innovation Competence, Humoral Immune Reactions in Cardiovascular Diseases, University Medicine Greifswald, 17489 Greifswald, Germany
b Institute for Immunology and Transfusion Medicine, University Medicine Greifswald, 17475 Greifswald, Germany
Knowledge about the mechanism by which some antibodies can block HIV-1 entry is critical to our understanding of their function and may offer new avenues for controlling the adhesion of HIV-1 to the host cells. While much progress has been made, this mechanism remains unclear. Here, atomic force microscopy, isothermal titration calorimetry (ITC), and circular dichroism spectroscopy were used to measure some biophysical characteristics of the interaction of four-domains (D1–D4) membrane protein CD4 with anti-D3 antibody OKT4 and with HIV-1 entry blocking anti-D1 antibody Leu3a. The results showed that at 37 °C they bind with similar binding strength, thermodynamics, and kinetics but with different assembly states. Further analyzing the interactions at different temperatures by ITC showed that binding of CD4 with Leu3a is characteristic for specific hydrophobic binding as well as for protein folding while with OKT4 comes from an extensive additional hydration upon binding and charge-related interactions within the binding site. Comparing these characteristics with those of HIV-1 gp120-CD4 interaction revealed that Leu3a binds to CD4 faster than HIV-1 followed by changing local structure of D1 to which HIV-1 binds leading to a prevention of viral entry.
Torification of diagonalizable group actions on toroidal schemes

Journal of Algebra, Available online 21 October 2016


Dan Abramovich (a), Michael Temkin (b)
a Department of Mathematics, Box 1917, Brown University, Providence, RI 02912, USA
b Einstein Institute of Mathematics, The Hebrew University of Jerusalem, Giv'at Ram, Jerusalem, 91904, Israel
We study actions of diagonalizable groups on toroidal schemes (i.e. logarithmically regular logarithmic schemes). In particular, we show that for so-called toroidal actions the quotient is again a toroidal scheme. Our main result constructs for an arbitrary action a canonical torification by an equivariant blowings up. This extends earlier results of Abramovich-de Jong, Abramovich-Karu-Matsuki-Włodarczyk, and Gabber in various aspects.

 

 July 2016

 

Title Abstract
Applications of imaging flow cytometry in the diagnostic assessment of acute leukaemia  

Methods, Available online 6 July 2016, Pages


Authors:
Lizz F. Grimwade a, Kathryn A. Fuller b, Wendy N. Erber b a Haemato-Oncology Diagnostic Service, Cambridge University Hospitals Foundation Trust, Cambridge CB2 0QQ, United Kingdom b Pathology and Laboratory Medicine, The University of Western Australia, Crawley, W.A. 6009, Australia
Automated imaging flow cytometry integrates flow cytometry with digital microscopy to produce high-resolution digital imaging with quantitative analysis. This enables cell identification based on morphology (cell size, shape), antigen expression, quantification of fluorescence signal intensity and localisation of detected signals (i.e. surface, cytoplasm, nuclear). We describe applications of imaging flow cytometry for the diagnostic assessment of acute leukaemia. These bone marrow malignancies are traditionally diagnosed and classified by cell morphology, phenotype and cytogenetic abnormalities. Traditionally morphology is assessed by light microscopy, phenotyping by conventional flow cytometry and genetics by karyotype and fluorescence in situ hybridisation (FISH) on interphase nuclei/metaphase spreads of cells on slides. Imaging flow cytometry adds a new dimension to the diagnostic assessment of these neoplasms. We describe three specific applications: 1) Assessment of PML bodies in acute promyelocytic leukaemia, 2) The nuclear and cytoplasmic localisation of NPM antigen in acute myeloid leukaemia, and, 3) The ability to detect cytogenetic abnormalities (i.e. aneuploidy) by automated FISH on intact whole cells in suspension. From this we conclude that imaging flow cytometry offers benefits over conventional diagnostic methods. Specifically the ability to visualise the cells of interest, the pattern and localisation of expressed antigens and assess cytogenetic abnormalities in one integrated automated high-throughput test. Imaging flow cytometry presents a new paradigm for the diagnostic assessment of leukaemia.
Imaging flow cytometry for the screening of compounds that disrupt the Plasmodium falciparum digestive vacuole   

Methods, Available online 5 July 2016, Pages

Authors:
Wan Ni Chiaa, Yan Quan Leea,b, Kevin Shyong-Wei Tana,b a Department of Microbiology and Immunology, National University of Singapore, Singapore b NUS Graduate School for Integrative Sciences and Engineering, National University of Singapore, Singapore
Malaria, despite being one of the world’s oldest infectious diseases, remains difficult to eradicate because the parasite is rapidly developing resistance to frontline chemotherapies. Previous studies have shown that the parasite exhibits features resembling programmed cell death upon treatment with drugs that disrupt its digestive vacuole (DV), providing a phenotypic readout that can be detected using the imaging flow cytometer. Large compound collections can thus be screened to identify drugs that are able to disrupt the DV of the malaria parasite using this high-content high-throughput screening platform. As a proof-of-concept, 4440 compounds were screened using this platform in 4 months and 254 hits (5.7% hit rate) were obtained. Additionally, 25 compounds (0.6% top hit rate) were observed to retain potent DV disruption activity that was comparable to the canonical DV disruptive drug chloroquine when tested at a ten-fold lower concentration from the original screen. This pilot study demonstrates the robustness and high-throughput capability of the imaging flow cytometer and we report herein the methodology of this screening assay.
Multi-parametric imaging of cell heterogeneity in apoptosis analysis  

Review Article
Methods, Available online 5 July 2016, Pages

Authors:
Ivan Vorobjeva,b,c, Natasha S Bartenevad,e a A.N. Belozersky Institute of Physico-Chemical Biology, M.V. Lomonosov Moscow State University, Russian Federation b Department of Cell Biology and Histology, M.V. Lomonosov Moscow State University, Russian Federation c School of Science and Technology, Nazarbayev University, Kazakhstan d Program in Cellular and Molecular Medicine, Boston Childrens Hospital, Harvard Medical School, MA, USA e Department of Pediatrics, Harvard Medical School, MA, USA
Apoptosis is a multistep process of programmed cell death where different morphological and molecular events occur simultaneously and/or consequently. Recent progress in programmed cell death analysis uncovered large heterogeneity in response of individual cells to the apoptotic stimuli. Analysis of the complex and dynamic process of apoptosis requires a capacity to quantitate multiparametric data obtained from multicolor labeling and/or fluorescent reporters of live cells in conjunction with morphological analysis. Modern methods of multiparametric apoptosis study include but are not limited to fluorescent microscopy, flow cytometry and imaging flow cytometry. In the current review we discuss the image-based evaluation of apoptosis on the single-cell and population level by imaging flow cytometry in parallel with other techniques. The advantage of imaging flow cytometry is its ability to interrogate multiparametric morphometric and fluorescence quantitative data in statistically robust manner. Here we describe the current status and future perspectives of this emerging field, as well as some challenges and limitations. We also highlight a number of assays and multicolor labeling probes, utilizing both microscopy and different variants of imaging cytometry, including commonly based assays and novel developments in the field.
Consumption of a high-fat, high-calorie meal is associated with an increase in intracellular co-localization of PPAR-γ mRNA and protein in monocytes   

Original Research Article
Methods, Available online 11 July 2016, Pages

Authors:
Andrea L. Henning, Brian K. McFarlin University of North Texas, Applied Physiology Laboratory, United States University of North Texas, Department of Biological Sciences, United States
Acute and habitual dietary habits contribute to the onset and progression of many forms of cardiovascular disease. Circulating peripheral blood monocytes have been a target of pre-clinical research related to the risk of atherosclerosis. Specifically, when monocytes migrate into the subendothelial space and endocytosize modified LDL (i.e. acLDL or oxLDL) they phenotypically transform into foam cells. The endocytosis of modified LDL is mediated by the scavenger receptor CD36, whose expression is in tern regulated by the transcription factor PPAR-γ. In this report, we describe a novel technique for the simultaneous measurement of intracellular PPAR-γ mRNA and protein in peripheral blood monocytes collected from human subjects in fasted state or 3 and 5-h after consuming a high-calorie (65% of daily calorie needs), high-fat meal. Intracellular detection and co-localization of PPAR-γ was made possible using a combination of image-based flow cytometry (MilliporeSigma FlowSight) and an amplified mRNA FISH staining technique (Affymetrix/eBioscience PrimeFlow). Consumption of a high-calorie, high-fat meal increased the percentage of co-localization at both 3 and 5-h post prandial compared to pre-meal. No obvious difference in co-localization was observed when cells were treated by acLDL in vitro. More research is needed to determine how to best use this method to study pre-clinical risk of atherosclerosis.
Characterization of extracellular vesicles in whole blood: Influence of pre-analytical parameters and visualization of vesicle-cell interactions using imaging flow cytometry   

Biochemical and Biophysical Research Communications, Available online 19 July 2016, Pages

Authors:
Birgit Fendl a, René Weiss a, Michael B. Fischer a, Andreas Spittler b, Viktoria Weber aa Christian Doppler Laboratory for Innovative Therapy Approaches in Sepsis, Department for Health Sciences and Biomedicine, Danube University Krems, Austria b Core Facility Flow Cytometry & Surgical Research Laboratories, Medical University of Vienna, Austria
Extracellular vesicles are central players in intercellular communication and are released from the plasma membrane under tightly regulated conditions, depending on the physiological and pathophysiological state of the producing cell. Their heterogeneity requires a spectrum of methods for isolation and characterization, where pre-analytical parameters have profound impact on vesicle analysis, particularly in blood, since sampling, addition of anticoagulants, as well as post-sampling vesicle generation may influence the outcome. Here, we characterized microvesicles directly in whole blood using a combination of flow cytometry and imaging flow cytometry. We assessed the influence of sample agitation, anticoagulation, and temperature on post-sampling vesicle generation, and show that vesicle counts remained stable over time in samples stored without agitation. Storage with gentle rolling mimicking agitation, in contrast, resulted in strong release of platelet-derived vesicles in blood anticoagulated with citrate or heparin, whereas vesicle counts remained stable upon anticoagulation with EDTA. Using imaging flow cytometry, we could visualize microvesicles adhering to blood cells and revealed an anticoagulant-dependent increase in vesicle-cell aggregates over time. We demonstrate that vesicles adhere preferentially to monocytes and granulocytes in whole blood, while no microvesicles could be visualized on lymphocytes. Our data underscore the relevance of pre-analytical parameters in vesicle analysis and demonstrate that imaging flow cytometry is a suitable tool to study the interaction of extracellular vesicles with their target cells.  
Radotinib inhibits acute myeloid leukemia cell proliferation via induction of mitochondrial-dependent apoptosis and CDK inhibitors   

European Journal of Pharmacology, Available online 28 July 2016, Pages


Authors:
Sook-Kyoung Heoa, Eui-Kyu Nohb, Gi-Dong Gwona, Jeong Yi Kima, Jae-Cheol Job, Yunsuk Choib, SuJin Kohb, Jin Ho Baekb, Young Joo Minb, Hawk Kima,b a Biomedical Research Center, Ulsan University Hospital, University of Ulsan College of Medicine, Ulsan 682-060, Republic of Korea b Department of Hematology and Oncology, Ulsan University Hospital, University of Ulsan College of Medicine, Ulsan 682-714, Republic of Korea
Radotinib is a BCR-ABL1 tyrosine kinase inhibitor approved for the second-line treatment of chronic myeloid leukemia. However, effects of radotinib on acute myeloid leukemia (AML) are unclear. In the present study, we observed that radotinib exerted cytotoxic effects on AML cells. Of the various AML cell lines examined (NB4, HL60, HEL 92.1.7, and THP-1), Kasumi-1 was the most sensitive to radotinib. Results of microarray analysis showed that 417 and 595 genes associated with apoptosis and cell cycle regulation, respectively, were differently expressed (i.e., showed >2-fold difference in expression). Radotinib-induced apoptosis involved the mitochondrial pathway. Moreover, radotinib increased the apoptosis of and induced caspase-3 activity in both Kasumi-1 cells and bone marrow cells (BMCs) obtained from patients with AML. Radotinib also increased cleaved caspase-3, caspase-7, and caspase-9 levels and decreased the number of proliferating Kasumi-1 cells and BMCs from patients with AML. In addition, radotinib induced G0/G1 phase arrest by inducing CDKIs p21 and p27 and by inhibiting CDK2, CDK4, and CDK6. These results indicate that radotinib induces caspase-dependent apoptosis and G0/G1 phase arrest in AML cells by regulating CDKI–CDK–cyclin cascade. Moreover, these results indicate that radotinib inhibits AML cell proliferation by inducing mitochondria-dependent apoptosis and CDKIs p21 and p27. To our knowledge, this is the first study to show that radotinib can be potentially used for the anti-leukemic therapy of patients with AML.
Masks in Imaging Flow Cytometry  

Methods, Available online 25 July 2016, Pages


Authors:
Venina Dominical, Leigh Samsel, J. Philip McCoy National Heart, Lung, and Blood Institute, NIH, Bethesda, MD 20892, United States
Data analysis in imaging flow cytometry incorporates elements of flow cytometry together with other aspects of morphological analysis of images. A crucial early step in this analysis is the creation of a mask to distinguish the portion of the image upon which further examination of specified features can be performed. Default masks are provided by the manufacturer of the imaging flow cytometer but additional custom masks can be created by the individual user for specific applications. Flawed or inaccurate masks can have a substantial negative impact on the overall analysis of a sample, thus great care must be taken to ensure the accuracy of masks. Here we discuss various types of masks and cite examples of their use. Furthermore we provide our insight for how to approach selecting and assessing the optimal mask for a specific analysis.