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https://www.readbyqxmd.com/read/28087633/the-abl-pathway-bifurcates-to-balance-enabled-and-rac-signaling-in-axon-patterning-in-drosophila
#1
Ramakrishnan Kannan, Jeong-Kuen Song, Tatiana Karpova, Akanni Clarke, Madhuri Shivalkar, Benjamin Wang, Lyudmila Kotlyanskaya, Irina Kuzina, Qun Gu, Edward Giniger
The Abl tyrosine kinase signaling network controls cell migration, epithelial organization, axon patterning and other aspects of development. While individual components are known, the relationships among them remain mysterious. We now use FRET measurements of pathway activity, analysis of protein localization and genetic epistasis to dissect the structure of this network in Drosophila We find that the adaptor protein Disabled stimulates Abl kinase activity. Abl suppresses the actin regulatory factor Enabled, and we find that Abl also acts through the GEF Trio to stimulate the signaling activity of Rac GTPase: Abl gates the activity of the spectrin repeats of Trio, allowing them to relieve intramolecular repression of Trio GEF activity by the Trio N-terminal domain...
January 13, 2017: Development
https://www.readbyqxmd.com/read/28081295/connected-component-masking-accurately-identifies-the-ratio-of-phagocytosed-and-cell-bound-particles-in-individual-cells-by-imaging-flow-cytometry
#2
Chenjie Fei, Dustin M E Lillico, Brian Hall, Aja M Rieger, James L Stafford
Innate immune cell-mediated recognition, capture, and engulfment of large particulate targets such as bacteria is known as phagocytosis. This highly dynamic cellular process involves a series of steps including receptor-mediated target binding, phagocytic cup formation, pseudopod extension, and phagosome closure, which depend on distinct actin polymerization events. Using flow cytometry, precise determination of target locations relative to cell membranes (i.e., surface-bound vs. fully engulfed/internalized) during the phagocytic process is difficult to quantify...
January 12, 2017: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/28081281/short-communication-validation-of-reference-genes-for-real-time-quantitative-pcr-in-tambaqui-colossoma-macropomum
#3
A R Nascimento, G F Silva, G F Gualberto, F L Almeida
Tambaqui, Colossoma macropomum, is the main native freshwater fish in Brazilian aquaculture. Therefore, intensive research pressure has been applied to the species to support new technologies for tambaqui farming. Molecular biology represents a tool that can be used to investigate every field of applied biology, from fish physiology to the effects of climate change. Based on the importance of reference genes for the relative or absolute quantification of gene transcripts, we cloned and sequenced three candidate reference genes in tambaqui (18S ribossomal RNA - 18s, glyceraldehyde-3-phosphate dehydrogenase - gapdh, and actin beta - β-actin), and validated a set of primers for each gene for use in real-time quantitative PCR...
December 23, 2016: Genetics and Molecular Research: GMR
https://www.readbyqxmd.com/read/28081156/generation-of-a-stable-transgenic-swine-model-expressing-a-porcine-histone-2b-egfp-fusion-protein-for-cell-tracking-and-chromosome-dynamics-studies
#4
Renan B Sper, Sehwon Koh, Xia Zhang, Sean Simpson, Bruce Collins, Jeff Sommer, Robert M Petters, Ignacio Caballero, Jeff L Platt, Jorge A Piedrahita
Transgenic pigs have become an attractive research model in the field of translational research, regenerative medicine, and stem cell therapy due to their anatomic, genetic and physiological similarities with humans. The development of fluorescent proteins as molecular tags has allowed investigators to track cell migration and engraftment levels after transplantation. Here we describe the development of two transgenic pig models via SCNT expressing a fusion protein composed of eGFP and porcine Histone 2B (pH2B)...
2017: PloS One
https://www.readbyqxmd.com/read/28081137/anillin-phosphorylation-controls-timely-membrane-association-and-successful-cytokinesis
#5
Hyunjung Kim, James M Johnson, Robert F Lera, Sarang Brahma, Mark E Burkard
During cytokinesis, a contractile ring generates the constricting force to divide a cell into two daughters. This ring is composed of filamentous actin and the motor protein myosin, along with additional structural and regulatory proteins, including anillin. Anillin is a required scaffold protein that links the actomyosin ring to membrane and its organizer, RhoA. However, the molecular basis for timely action of anillin at cytokinesis remains obscure. Here, we find that phosphorylation regulates efficient recruitment of human anillin to the equatorial membrane...
January 2017: PLoS Genetics
https://www.readbyqxmd.com/read/28079575/cystic-biliary-atresia-and-choledochal-cysts-are-distinct-histopathologic-entities
#6
Inna N Lobeck, Rachel Sheridan, Mark Lovell, Phylicia Dupree, Greg M Tiao, Kevin E Bove
Cystic biliary atresia (CBA), a rare cystic expansion of atretic extrahepatic bile ducts in young infants, overlaps in age at presentation and imaging features with early choledochal cysts (CC). Treatment and prognosis differ; histologic differences are unsettled. We compared 10 patients with CBA, 1975 to 2015, to an age-similar cohort of 13 infants, and to older patients who had surgery for CC. Operative details, imaging, and clinical courses were correlated to pathologic specimens. Immunostains for smooth muscle actin and myosin heavy chain were used to evaluate cyst walls and atretic segments...
January 10, 2017: American Journal of Surgical Pathology
https://www.readbyqxmd.com/read/28078820/improvement-of-the-in-vivo-cellular-repopulation-of-decellularized-cardiovascular-tissues-by-a-detergent-free-non-proteolytic-actin-disassembling-regimen
#7
Alexander Assmann, Marc Struß, Franziska Schiffer, Friederike Heidelberg, Hiroshi Munakata, Elena V Timchenko, Pavel E Timchenko, Tim Kaufmann, Khon Huynh, Yukiharu Sugimura, Quentin Leidl, Antonio Pinto, Volker R Stoldt, Artur Lichtenberg, Payam Akhyari
Low immunogenicity and high repopulation capacity are crucial determinants for the functional and structural performance of acellular cardiovascular implants. The present study evaluates a detergent-free, non-proteolytic, actin-disassembling regimen (BIO) for decellularization of heart valve and vessel grafts, particularly focusing on their bio-functionality. Rat aortic conduits (rAoC; n = 89) and porcine aortic valve samples (n = 106) are decellularized using detergents (group DET) or the BIO regimen...
January 11, 2017: Journal of Tissue Engineering and Regenerative Medicine
https://www.readbyqxmd.com/read/28078746/a-gly65val-substitution-in-an-actin-ghact_li1-disrupts-cell-polarity-and-f-actin-organization-resulting-in-dwarf-lintless-cotton-plants
#8
Gregory N Thyssen, David D Fang, Rickie B Turley, Christopher B Florane, Ping Li, Christopher P Mattison, Marina Naoumkina
Actin polymerizes to form part of the cytoskeleton and organize polar growth in all eukaryotic cells. Species with numerous actin genes are especially useful for the dissection of actin molecular function due to redundancy and neofunctionalization. Here, we investigated the role of a cotton (Gossypium hirsutum) actin gene in the organization of actin filaments in lobed cotyledon pavement cells and the highly-elongated single-celled trichomes that comprise cotton lint fibers. Using mapping-by-sequencing, virus-induced gene silencing, and molecular modeling, we identified the causative mutation of the dominant dwarf Ligon-lintless Li1 short fiber mutant as a single Gly65Val amino acid substitution in a polymerization domain of an actin gene, GhACT_LI1 (Gh_D04G0865)...
January 12, 2017: Plant Journal: for Cell and Molecular Biology
https://www.readbyqxmd.com/read/28078713/the-novel-mtor-complex-1-2-inhibitor-p529-inhibits-human-lung-myofibroblast-differentiation
#9
Keith T Ferguson, Elizabeth E Torr, Ksenija Bernau, Jonathan Leet, Davis Sherris, Nathan Sandbo
Idiopathic pulmonary fibrosis is a progressive and deadly disorder with very few therapeutic options. Palomid 529 (8-(1-hydroxyethyl)-2-methoxy-3-(4-methoxybenzyloxy)-benzo[c]chromen-6-one; P529) is a novel dual inhibitor of mechanistic target of rapamycin complex 1/2 (mTORC1/2). In these studies, we investigated the effect of P529 on TGF-β-dependent signaling and myofibroblast differentiation. TGF-β-induced phosphorylation of the mTORC1 targets, p70 S6 kinase 1 (S6K1) and eukaryotic translation initiation factor 4E binding protein 1 (4E-BP1), were both dose dependently inhibited by P529 in human lung fibroblasts with maximal inhibition occurring between 10-20 µM...
January 11, 2017: Journal of Cellular Biochemistry
https://www.readbyqxmd.com/read/28078678/a-seamless-phase-i-ii-dose-finding-trial-assessing-ingenol-disoxate-leo-43204-for-field-treatment-of-actinic-keratosis-on-the-scalp
#10
J Weiss, M Ulrich, M Bukhalo, M L Østerdal, B Bang, C W Hanke
BACKGROUND: Actinic keratosis (AK) is a common sun-related skin condition, which can progress to squamous cell carcinoma and occur in cancerised fields. OBJECTIVES: This Phase I/II trial investigated the safety and efficacy of ingenol disoxate as topical field therapy for patients with AK on the balding scalp. METHODS: Part 1 was a Phase I, open-label, dose-escalation trial investigating up to 6 doses of ingenol disoxate to determine the maximum tolerated dose (MTD)...
January 12, 2017: British Journal of Dermatology
https://www.readbyqxmd.com/read/28078583/generation-and-application-of-bioluminescent-cd95-ligand-fusion-proteins
#11
Isabell Lang, Juliane Kums, Harald Wajant
The quantitative evaluation of the interaction of soluble CD95L with CD95 is not only important for a detailed understanding of CD95 biology but is also of special relevance for the characterization and development of inhibitors of this interaction. The assembly of a CD95L-CD95 complex capable to recruit intracellular factors not only involves pre-assembly of CD95 molecules in the absence of CD95L but is also modulated by cellular factors such as interaction with the actin cytoskeleton and plasma membrane compartmentation of CD95...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28078480/muscarinic-acetylcholine-receptor-mediated-tight-junction-opening-is-involved-in-epiphora-in-late-phase-of-submandibular-gland-transplantation
#12
Ning-Yan Yang, Chong Ding, Jing Li, Yan Zhang, Ruo-Lan Xiang, Li-Ling Wu, Guang-Yan Yu, Xin Cong
Submandibular gland (SMG) autotransplantation is an effective therapy for treating severe dry eye syndrome. However, epiphora occurs in more than 40% of patients 6 months after operation. We previously found that muscarinic acetylcholine receptor (mAChR) plays a crucial role in regulating SMG secretion partially through the modulation on tight junction (TJ)-based paracellular pathway. Therefore, the present study aimed to investigate the possible involvement of mAChR and TJ in a rabbit long-term model of SMG transplantation...
January 11, 2017: Journal of Molecular Histology
https://www.readbyqxmd.com/read/28078426/calreticulin-is-required-for-calcium-homeostasis-and-proper-pollen-tube-tip-growth-in-petunia
#13
Anna Suwińska, Piotr Wasąg, Przemysław Zakrzewski, Marta Lenartowska, Robert Lenartowski
Calreticulin is involved in stabilization of the tip-focused Ca (2+) gradient and the actin cytoskeleton arrangement and function that is required for several key processes driving Petunia pollen tube tip growth. Although the precise mechanism is unclear, stabilization of a tip-focused calcium (Ca(2+)) gradient seems to be critical for pollen germination and pollen tube growth. We hypothesize that calreticulin (CRT), a Ca(2+)-binding/buffering chaperone typically residing in the lumen of the endoplasmic reticulum (ER) of eukaryotic cells, is an excellent candidate to fulfill this role...
January 11, 2017: Planta
https://www.readbyqxmd.com/read/28077619/relative-importance-of-%C3%AE-cyto-and-%C3%AE-cyto-actin-in-primary-mouse-embryonic-fibroblasts
#14
Xiaobai Patrinostro, Allison R O'Rourke, Christopher M Chamberlain, Branden S Moriarity, Benjamin J Perrin, James M Ervasti
The highly homologous beta (βcyto) and gamma (γcyto) cytoplasmic actins are hypothesized to carry out both redundant and unique essential functions, but studies using targeted gene knockout and siRNA mediated transcript knockdown to examine βcyto- and γcyto-isoform specific functions in various cell types have yielded conflicting data. Here, we quantitatively characterized actin transcript and protein levels as well as cellular phenotypes in both gene- and transcript-targeted primary mouse embryonic fibroblasts...
January 11, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/28077579/neuron-enriched-rna-binding-proteins-regulate-pancreatic-beta-cell-function-and-survival
#15
Jon Agraves Juan-Mateu, Tatiana H Rech, Olatz Villate, Esther Lizarraga-Mollinedo, Anna Wendt, Jean-Valery Turatsinze, Leticia A Brondani, Tarlliza R Nardelli, Tatiane C Nogueira, Jonatan L S Esguerra, Maria In Ecircs Alvelos, Piero Marchetti, Lena Eliasson, D Eacutecio L Eizirik
Pancreatic beta cell failure is the central event leading to diabetes. Beta cells share many phenotypic traits with neurons, and proper beta cell function relies on the activation of several neuronal-like transcription programs. Regulation of gene expression by alternative splicing plays a pivotal role in brain, where it affects neuronal development, function and disease. The role of alternative splicing in beta cells remains unclear, but recent data indicate that splicing alterations modulated by both inflammation and susceptibility genes for diabetes contribute to beta cell dysfunction and death...
January 11, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28077511/altered-connectivity-and-synapse-maturation-of-the-hippocampal-mossy-fiber-pathway-in-a-mouse-model-of-the-fragile-x-syndrome
#16
F Scharkowski, Michael Frotscher, David Lutz, Martin Korte, Kristin Michaelsen-Preusse
The Fragile X syndrome (FXS) as the most common monogenetic cause of cognitive impairment and autism indicates how tightly the dysregulation of synapse development is linked to cognitive deficits. Symptoms of FXS include excessive adherence to patterns that point to compromised hippocampal network formation. Surprisingly, one of the most complex hippocampal synapses connecting the dentate gyrus (DG) to CA3 pyramidal neurons has not been analyzed in FXS yet. Intriguingly, we found altered synaptic function between DG and CA3 in a mouse model of FXS (fmr1 knockout [KO]) demonstrated by increased mossy fiber-dependent miniature excitatory postsynaptic current (mEPSC) frequency at CA3 pyramidal neurons together with increased connectivity between granule cells and CA3 neurons...
January 10, 2017: Cerebral Cortex
https://www.readbyqxmd.com/read/28077319/role-of-insulin-like-growth-factor-axis-in-the-bleomycin-induced-lung-injury-in-rats
#17
Lakshmi Kanth Kotarkonda, Ritu Kulshrestha, Krishnan Ravi
BACKGROUND: Alveolar epithelial cell injury has been proposed as a causative factor for the onset and progression ofpulmonary fibrosis. However, the role of type II alveolar epithelial cells (AECs) in the epithelial mesenchymal transition (EMT) is controversial. AIMS: The present study performed in rats instilled with bleomycin investigated a) the expressions of the insulin growth factor (IGF-1) and insulin growth factor binding protein 5 (IGFBP-5) and transforming growth factor (TGF-β1) in the type II AECs, b) the role oftype II AECs in EMT andextracellular matrix (ECM) formation and, c) the effect of pioglitazone on all the above parameters...
January 7, 2017: Experimental and Molecular Pathology
https://www.readbyqxmd.com/read/28077285/a-fluorophore-fusion-construct-of-human-profilin-i-with-non-compromized-poly-l-proline-binding-capacity-suitable-for-imaging
#18
Michaela Nejedla, Zhilun Li, Anna E Masser, Matteo Biancospino, Matthias Spiess, Sebastian D Mackowiak, Marc R Friedländer, Roger Karlsson
Profilin is vital for actin organization in eukaryotic cells. It controls actin filament formation by binding monomeric actin and numerous proteins involved in polarized actin assembly. Important for the latter is the interaction surface formed by the N- and C-terminal helices, which pack close to each other on one side of the molecule at distance from the actin site and mediate binding to poly-proline sequences present in many of the targeted proteins. Via these interactions profilin contributes to the spatiotemporal control of actin filament growth...
January 7, 2017: Journal of Molecular Biology
https://www.readbyqxmd.com/read/28077174/mutant-hspb1-causes-loss-of-translational-repression-by-binding-to-pcbp1-an-rna-binding-protein-with-a-possible-role-in-neurodegenerative-disease
#19
Thomas Geuens, Vicky De Winter, Nicholas Rajan, Tilmann Achsel, Ligia Mateiu, Leonardo Almeida-Souza, Bob Asselbergh, Delphine Bouhy, Michaela Auer-Grumbach, Claudia Bagni, Vincent Timmerman
The small heat shock protein HSPB1 (Hsp27) is an ubiquitously expressed molecular chaperone able to regulate various cellular functions like actin dynamics, oxidative stress regulation and anti-apoptosis. So far disease causing mutations in HSPB1 have been associated with neurodegenerative diseases such as distal hereditary motor neuropathy, Charcot-Marie-Tooth disease and amyotrophic lateral sclerosis. Most mutations in HSPB1 target its highly conserved α-crystallin domain, while other mutations affect the C- or N-terminal regions or its promotor...
January 11, 2017: Acta Neuropathologica Communications
https://www.readbyqxmd.com/read/28076931/varying-effects-of-egf-hgf-and-tgf%C3%AE-on-formation-of-invadopodia-and-invasiveness-of-melanoma-cell-lines-of-different-origin
#20
A Makowiecka, A Simiczyjew, D Nowak, A J Mazur
The understanding of melanoma malignancy mechanisms is essential for patient survival, because melanoma is responsible for ca. 75% of deaths related to skin cancers. Enhanced formation of invadopodia and extracellular matrix (ECM) degradation are two important drivers of cell invasion, and actin dynamics facilitate protrusive activity by providing a driving force to push through the ECM. We focused on the influence of epidermal growth factor (EGF), hepatocyte growth factor (HGF) and transforming growth factor β (TGFβ) on melanoma cell invasiveness, since they are observed in the melanoma microenvironment...
December 9, 2016: European Journal of Histochemistry: EJH
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