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  • 1
    Language: English
    In: Neuroscience and Behavioral Physiology, 2015, Vol.45(9), pp.1077-1081
    Description: Studies of the correlation between neuron activity and behavior require objective measurement of animal movement. This is obtained using video recordings followed by analysis of video images. The conditions for recording neuron activity in a Morris basin require the video camera to be positioned at an angle to the water surface, which leads to perspective distortion of the images and, as a result, errors in trajectory analysis. In the present study we present a method for correcting perspective distortions.
    Keywords: rats ; Morris basin ; neuron activity ; video tracking ; perspective distortions
    ISSN: 0097-0549
    E-ISSN: 1573-899X
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  • 2
    In: Circulation, 2012, Vol.126(20), pp.2418-2427
    Description: BACKGROUND—: Carotid intima-media thickening is associated with increased cardiovascular risk in humans. We discovered that intima formation and cell proliferation in response to carotid injury is greater in SJL/J (SJL) in comparison with C3HeB/FeJ (C3H/F) mice. The purpose of this study was to identify candidate genes contributing to intima formation. METHODS AND RESULTS—: We performed microarray and bioinformatic analyses of carotid arteries from C3H/F and SJL mice. Kyoto Encyclopedia of Genes and Genomes analysis showed that the ribosome pathway was significantly up-regulated in C3H/F in comparison with SJL mice. Expression of a ribosomal protein, RpL17, was 〉40-fold higher in C3H/F carotids in comparison with SJL. Aortic vascular smooth muscle cells from C3H/F grew slower in comparison to SJL. To determine the role of RpL17 in vascular smooth muscle cell growth regulation, we analyzed the relationship between RpL17 expression and cell cycle progression. Cultured vascular smooth muscle cells from mice, rats, and humans showed that RpL17 expression inversely correlated with growth as shown by decreased cells in S phase and increased cells in G0/G1. To prove that RpL17 acted as a growth inhibitor in vivo, we used pluronic gel delivery of RpL17 small interfering RNA to C3H/F carotid arteries. This resulted in an 8-fold increase in the number of proliferating cells. Furthermore, following partial carotid ligation in SJL mice, RpL17 expression in the intima and media decreased, but the number of proliferating cells increased. CONCLUSIONS—: RpL17 acts as a vascular smooth muscle cell growth inhibitor (akin to a tumor suppressor) and represents a potential therapeutic target to limit carotid intima-media thickening.
    Keywords: Medicine ; Anatomy & Physiology;
    ISSN: 0009-7322
    E-ISSN: 15244539
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  • 3
    In: Nature, 2013, Vol.497(7451), p.624
    Description: Recent exon-sequencing studies of human tumours have revealed that subunits of BAF (mammalian SWI/SNF) complexes are mutated in more than 20% of all human malignancies, but the mechanisms involved in tumour suppression are unclear. BAF chromatin-remodelling complexes are polymorphic assemblies that use energy provided by ATP hydrolysis to regulate transcription through the control of chromatin structure and the placement of Polycomb repressive complex 2 (PRC2) across the genome. Several proteins dedicated to this multisubunit complex, including BRG1 (also known as SMARCA4) and BAF250a (also known as ARID1A), are mutated at frequencies similar to those of recognized tumour suppressors. In particular, the core ATPase BRG1 is mutated in 5-10% of childhood medulloblastomas and more than 15% of Burkitt's lymphomas. Here we show a previously unknown function of BAF complexes in decatenating newly replicated sister chromatids, a requirement for proper chromosome segregation during mitosis. We find that deletion of Brg1 in mouse cells, as well as the expression of BRG1 point mutants identified in human tumours, leads to anaphase bridge formation (in which sister chromatids are linked by catenated strands of DNA) and a G2/M-phase block characteristic of the decatenation checkpoint. Endogenous BAF complexes interact directly with endogenous topoisomerase IIα (TOP2A) through BAF250a and are required for the binding of TOP2A to approximately 12,000 sites across the genome. Our results demonstrate that TOP2A chromatin binding is dependent on the ATPase activity of BRG1, which is compromised in oncogenic BRG1 mutants. These studies indicate that the ability of TOP2A to prevent DNA entanglement at mitosis requires BAF complexes and suggest that this activity contributes to the role of BAF subunits as tumour suppressors.
    Keywords: Antigens, Neoplasm -- Metabolism ; DNA Helicases -- Metabolism ; DNA Topoisomerases, Type II -- Metabolism ; DNA, Catenated -- Chemistry ; DNA-Binding Proteins -- Metabolism ; Nuclear Proteins -- Metabolism ; Transcription Factors -- Metabolism;
    ISSN: 0028-0836
    E-ISSN: 14764687
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  • 4
    Language: English
    In: Journal of Neuroscience Methods, 30 April 2012, Vol.206(1), pp.15-22
    Description: ► We described the multichannel preamplifier for neuronal recordings in small animals. ► The device may be used in any kinds of tasks including swimming in Morris water maze. ► Reusable headstage can adopt both chronically implanted and movable wire electrodes. ► For the first time multichannel recordings of neuronal activity were made in swimming mice. The design of a miniature multichannel preamplifier for extracellular recordings of single unit activity in freely moving and swimming small animals is presented. The advantages of this design include perfect protection of the critical components and electric contacts from water. Thus, neuronal activity and EEG may be recorded differentially in any kinds of behavioral tasks including swimming in Morris water maze. Recordings are stable even if an animal is diving and swimming under the water surface. The reusable dismountable base can adopt different types of chronically implanted fine wire electrodes and movable arrays. Electrodes may be implanted to any desired depth. The assembly weight is less than 240 mg. Thus, the construction is light enough even for mice. This work is the first successful attempt for multichannel recording of neuronal activity in mice performing spatial task in Morris water maze.
    Keywords: Multichannel Preamplifier ; Reusable Base ; Microdrive ; Fine Wire Electrodes ; Single Unit Activity ; Freely Moving Animals ; Swimming Mice and Rats ; Water Maze ; Medicine ; Anatomy & Physiology
    ISSN: 0165-0270
    E-ISSN: 1872-678X
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  • 5
    Language: English
    In: Clinical science (London, England : 1979), April 2012, Vol.122(8), pp.361-8
    Description: Axl is a receptor tyrosine kinase that was originally cloned from cancer cells. Axl belongs to the TAM (Tyro3, Axl and Mertk) family of receptor tyrosine kinases. Gas6 (growth-arrest-specific protein 6) is a ligand for Axl. Activation of Axl protects cells from apoptosis, and increases migration, aggregation and growth through multiple downstream pathways. Up-regulation of the Gas6/Axl pathway is more evident in pathological conditions compared with normal physiology. Recent advances in Axl receptor biology are summarized in the present review. The emphasis is given to translational aspects of Axl-dependent signalling under pathological conditions. In particular, inhibition of Axl reduces tumorigenesis and prevents metastasis as well. Axl-dependent signals are important for the progression of cardiovascular diseases. In contrast, deficiency of Axl in innate immune cells contributes to the pathogenesis of autoimmune disorders. Current challenges in Axl biology are related to the functional interactions of Axl with other members of the TAM family or other tyrosine kinases, mechanisms of ligand-independent activation, inactivation of the receptor and cell-cell interactions (with respect to immune cells) in chronic diseases.
    Keywords: Signal Transduction ; Proto-Oncogene Proteins -- Metabolism ; Receptor Protein-Tyrosine Kinases -- Metabolism
    ISSN: 01435221
    E-ISSN: 1470-8736
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  • 6
    Language: English
    In: Proceedings of the National Academy of Sciences of the United States of America, 27 March 2018, Vol.115(13), pp.3392-3397
    Description: The main cell of origin of the Sonic hedgehog (SHH) subgroup of medulloblastoma (MB) is granule cell precursors (GCPs), a SHH-dependent transient amplifying population in the developing cerebellum. SHH-MBs can be further subdivided based on molecular and clinical parameters, as well as location because SHH-MBs occur preferentially in the lateral cerebellum (hemispheres). Our analysis of adult patient data suggests that tumors with Smoothened () mutations form more specifically in the hemispheres than those with Patched 1 () mutations. Using sporadic mouse models of SHH-MB with the two mutations commonly seen in adult MB, constitutive activation of () or loss-of-, we found that regardless of timing of induction or type of mutation, tumors developed primarily in the hemispheres, with -mutants indeed showing a stronger specificity. We further uncovered that GCPs in the hemispheres are more susceptible to high-level SHH signaling compared with GCPs in the medial cerebellum (vermis), as more or -mutant hemisphere cells remain undifferentiated and show increased tumorigenicity when transplanted. Finally, we identified location-specific GCP gene-expression profiles, and found that deletion of the genes most highly expressed in the hemispheres () or vermis (Engrailed1) showed opposing effects on GCP differentiation. Our studies thus provide insights into intrinsic differences within GCPs that impact on SHH-MB progression.
    Keywords: En1 ; Mri ; Nr2f2 ; Cerebellar Hemispheres ; Granule Cell Precursors ; Cerebellar Neoplasms -- Pathology ; Cerebellum -- Pathology ; Hedgehog Proteins -- Metabolism ; Medulloblastoma -- Pathology ; Patched-1 Receptor -- Metabolism ; Smoothened Receptor -- Metabolism
    ISSN: 00278424
    E-ISSN: 1091-6490
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  • 7
    Language: English
    In: The Journal of clinical investigation, April 2011, Vol.121(4), pp.1344-8
    Description: Pilocytic astrocytoma (PA) is the most common type of primary brain tumor in children and the second most frequent cancer in childhood. Children with incompletely resected PA represent a clinically challenging patient cohort for whom conventional adjuvant therapies are only moderately effective. This has produced high clinical demand for testing of new molecularly targeted treatments. However, the development of new therapeutics for PA has been hampered by the lack of an adequate in vivo tumor model. Recent studies have identified activation of MAPK signaling, mainly by oncogenic BRAF activation, as a hallmark genetic event in the pathogenesis of human PA. Using in vivo retroviral somatic gene transfer into mouse neural progenitor cells, we have shown here that ectopic expression of the activated BRAF kinase domain is sufficient to induce PA in mice. Further in vitro analyses demonstrated that overexpression of activated BRAF led to increased proliferation of primary mouse astrocytes that could be inhibited by treatment with the kinase inhibitor sorafenib. Our in vivo model for PA shows that the activated BRAF kinase domain is sufficient to induce PA and highlights its role as a potential therapeutic target.
    Keywords: Astrocytoma -- Etiology ; Brain Neoplasms -- Etiology ; Proto-Oncogene Proteins B-Raf -- Genetics
    ISSN: 00219738
    E-ISSN: 1558-8238
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  • 8
    Language: English
    In: Bulletin of Experimental Biology and Medicine, 2018, Vol.165(5), pp.695-697
    Description: Antitumor effects of glycolysis inhibitors monoiodoacetate and 2-deoxyglucose were studied on Lewis lung carcinoma model. Monoiodoacetate exhibited antitumor and antimetastatic activities, being not inferior of methotrexate (reference drug); however, the preparation also demonstrated high systemic toxicity. 2-Deoxyglucose exhibited only antitumor effect, while its antimetastatic activity did not differ from the result in the group without treatment.
    Keywords: Warburg effect ; glycolysis inhibitors ; tumor
    ISSN: 0007-4888
    E-ISSN: 1573-8221
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  • 9
    Language: English
    In: Vascular Pharmacology, September 2010, Vol.53(3-4), pp.185-192
    Description: We previously demonstrated that reactive oxygen species (ROS) activate Axl, a receptor tyrosine kinase, resulting in increased survival of rat aortic smooth muscle cells (RASMs). Our experiments in Axl knockout mice showed significant reduction in vascular pathologies. We hypothesize that selective pharmacological inhibitors of Axl could prove beneficial in treating vascular diseases associated with oxidative stress. We investigated a role for two novel compounds specific for Axl (R428 and R572) on ligand independent activation of Axl mediated cell survival and migration. Stimulation of RASMs with H O for 5 min significantly increased Akt phosphorylation (p-Akt). Inhibition at 50% (IC ) of p-Akt was calculated at lower concentrations in R428 (100 nM) and R572 (10 nM) compared to Fc-Axl (2 μg/mL). Flow cytometry staining with Annexin V showed a 2-fold increase in apoptosis with R428 and R572 compared to Fc-Axl after H O , which was validated by concomitant increases in cleaved caspase-3. Pretreatment with R428 and R572 decreased cell migration by ~ 50% in response to 20% serum (similar to that after Fc-Axl). R428 and R572 decreased intracellular production of ROS in comparison to Fc-Axl. In conclusion, R428 and R572 are more potent inhibitors of ligand independent mediated Axl signaling compared to Fc-Axl in RASMs under oxidative stress.
    Keywords: Receptor Tyrosine Kinase ; Axl ; Reactive Oxygen Species ; Apoptosis ; Migration ; Pharmacy, Therapeutics, & Pharmacology
    ISSN: 1537-1891
    E-ISSN: 1879-3649
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  • 10
    Language: English
    In: Blood, 23 May 2013, Vol.121(21), pp.4417-27
    Description: Activated platelets release many inflammatory molecules with important roles in accelerating vascular inflammation. Much is known about platelet and platelet-derived mediator interactions with endothelial cells and leukocytes, but few studies have examined the effects of platelets on components of the vascular wall. Vascular smooth muscle cells (VSMCs) undergo phenotypic changes in response to injury including the production of inflammatory molecules, cell proliferation, cell migration, and a decline in the expression of differentiation markers. In this study, we demonstrate that the platelet-derived chemokine platelet factor 4 (PF4/CXCL4) stimulates VSMC injury responses both in vitro and in vivo in a mouse carotid ligation model. PF4 drives a VSMC inflammatory phenotype including a decline in differentiation markers, increased cytokine production, and cell proliferation. We also demonstrate that PF4 effects are mediated, in part, through increased expression of the transcription factor Krüppel-like factor 4. Our data indicate an important mechanistic role for platelets and PF4 in VSMC injury responses both in vitro and in vivo.
    Keywords: Blood Platelets -- Immunology ; Carotid Artery Injuries -- Immunology ; Muscle, Smooth, Vascular -- Immunology ; Platelet Factor 4 -- Metabolism ; Vasculitis -- Immunology
    ISSN: 00064971
    E-ISSN: 1528-0020
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