Publications – Neuroscience Initiative

2022 Publications

X. Chen, A. Momin, S. Wanggou, X. Wang, H.-K. Min, W. Dou, Z. Gong, J. Chan, W. Dong, J. J. Fan, Y. Xiong, K. Talipova, H. Zhao, Y. X. Chen, K. Veerasammy, A. Fekete, S. A. Kumar, H. Liu, Q. Yang, J. E. Son, et al. Mechanosensitive Brain Tumor Cells Construct Blood-Tumor Barrier to Mask Chemosensitivity. Neuron, 2022. DOI: https://doi.org/10.1016/j.neuron.2022.10.007.

G. A. Soliman, R. R. Abzalimov, Y. He. MTORC1 and MTORC2 Complexes Regulate the Untargeted Metabolomics and Amino Acid Metabolites Profile through Mitochondrial Bioenergetic Functions in Pancreatic Beta Cells. Nutrients, 2022, 14 (15), 3022. DOI: https://doi.org/10.3390/nu14153022.

Y. X. Chen, K. Veerasammy, J. Yin, T. Choetso, T. Zhong, M. A. Choudhury, C. Weng, E. Xu, M. A. Hein, R. Abzalimov, Y. He. Sample Preparation for Rapid Lipid Analysis in Drosophila Brain Using Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry Imaging. JoVE (Journal of Visualized Experiments), 2022, No. 185, e63930. DOI: https://doi.org/10.3791/63930.

J. Buthmann, D. Huang, P. Casaccia, S. O’Neill, Y. Nomura, J. Liu. Prenatal Exposure to a Climate-Related Disaster Results in Changes of the Placental Transcriptome and Infant Temperament. Frontiers in Genetics, 2022, 13.

H.-J. Park, E. Tsai, D. Huang, M. Weaver, L. Frick, A. Alcantara, J. J. Moran, J. Patzig, C. V. Melendez-Vasquez, G. R. Crabtree, M. L. Feltri, J. Svaren, P. Casaccia. ACTL6a Coordinates Axonal Caliber Recognition and Myelination in the Peripheral Nerve. iScience, 2022, 25 (4), 104132. DOI: https://doi.org/10.1016/j.isci.2022.104132.

D. K. Dansu, J. Liang, I. Selcen, H. Zheng, D. F. Moore, P. Casaccia. PRMT5 Interacting Partners and Substrates in Oligodendrocyte Lineage Cells. Front. Cell. Neurosci., 2022, 16, 820226. DOI: https://doi.org/10.3389/fncel.2022.820226.

F. Cathomas, L. M. Holt, E. M. Parise, J. Liu, J. W. Murrough, P. Casaccia, E. J. Nestler, S. J. Russo. Beyond the Neuron: Role of Non-Neuronal Cells in Stress Disorders. Neuron, 2022, S0896-6273(22)00102-7. DOI: https://doi.org/10.1016/j.neuron.2022.01.033.

H. W. Korsmo, B. Dave, S. Trasino, A. Saxena, J. Liu, J. M. Caviglia, K. Edwards, M. Dembitzer, S. Sheeraz, S. Khaldi, X. Jiang. Maternal Choline Supplementation and High-Fat Feeding Interact to Influence DNA Methylation in Offspring in a Time-Specific Manner. Frontiers in Nutrition, 2022, 9.

Publications Archives

2021

A. Ntranos, H. J. Park, M. Wentling, V. Tolstikov, M. Amatruda, B. Inbar, S. Kim-Schulze, C. Frazier, J. Button, M. A. Kiebish, F. Lublin, K. Edwards, P. Casaccia. Bacterial Neurotoxic Metabolites in Multiple Sclerosis Cerebrospinal Fluid and Plasma. Brain, 2021, awab320. DOI: https://doi.org/10.1093/brain/awab320. Video Abstract
T. Roostaei, H.-U. Klein, Y. Ma, D. Felsky, P. Kivisäkk, S. M. Connor, A. Kroshilina, C. Yung, B. J. Kaskow, X. Shao, B. Rhead, J. M. Ordovás, D. M. Absher, D. K. Arnett, J. Liu, N. Patsopoulos, L. F. Barcellos, H. L. Weiner, P. L. De Jager. Proximal and Distal Effects of Genetic Susceptibility to Multiple Sclerosis on the T Cell Epigenome. Nat Commun, 2021, 12 (1), 7078. DOI: https://doi.org/10.1038/s41467-021-27427-w.
D. Marechal, D. K. Dansu, K. Castro, J. Patzig, L. Magri, B. Inbar, M. Gacias, S. Moyon, P. Casaccia. N-Myc Downstream Regulated Family Member 1 (NDRG1) Is Enriched in Myelinating Oligodendrocytes and Impacts Myelin Degradation in Response to Demyelination. Glia. 2021 Oct 23. DOI: https://doi.org/10.1002/glia.24108.
R.H. Huang, N. Nayeem, Y. He, J. Morales, D. Graham, R. Klajn, M. Contel, S. O’Brien, and R.V. Ulijn. Self‐Complementary Zwitterionic Peptides Direct Nanoparticle Assembly and Enable Enzymatic Selection of Endocytic Pathways, Adv. Mater., 2021, 2104962, DOI: https://doi.org/10.1002/adma.202104962.
F. Li, T. Y. Lo, L. Miles, Q. Wang, H. N. Noristani, D. Li, J. Niu, S. Trombley, J. I. Goldshteyn, C. Wang, S. Wang, J. Qiu, K. Pogoda, K. Mandal, M. Brewster, P. Rompolas, Y. He, P. A. Janmey, G. M. Thomas, S. Li, Y. Song. The Atr-Chek1 Pathway Inhibits Axon Regeneration in Response to Piezo-Dependent Mechanosensation. Nat Commun, 2021, 12 (1), 3845. DOI: https://doi.org/10.1038/s41467-021-24131-7.
S. Moyon, R. Frawley, D. Marechal, D. Huang, K. L. H. Marshall-Phelps, L. Kegel, S. M. K. Bøstrand, B. Sadowski, Y.-H. Jiang, D. A. Lyons, W. Möbius, P. Casaccia. TET1-Mediated DNA Hydroxymethylation Regulates Adult Remyelination in Mice. Nat Commun, 2021, 12 (1), 3359. DOI: https://doi.org/10.1038/s41467-021-23735-3.
D. Chang, Q. Brown, G. Tsui, Y. He, J. Liu, L. Shi, A. Rodríguez-Contreras. Distinct Cellular Profiles of Hif1a and Vegf MRNA Localization in Microglia, Astrocytes and Neurons during a Period of Vascular Maturation in the Auditory Brainstem of Neonate Rats. Brain Sciences, 2021, 11 (7), 944. DOI: https://doi.org/10.3390/brainsci11070944.
Daniel Eskenazi, Lauren Malave, Susana Mingote, Leora Yetnikoff, Samira Ztaou, Vlad Velicu, Stephen Rayport, Nao Chuhma, Dopamine neurons that cotransmit glutamate, from synapses to circuits to behavior, Frontiers in Neural Circuits vol. 15, 2021. doi: 10.3389/fncir.2021.665386
J. Yin, E. Spillman, E. S. Cheng, J. Short, Y. Chen, J. Lei, M. Gibbs, J. S. Rosenthal, C. Sheng, Y. X. Chen, K. Veerasammy, T. Choetso, R. Abzalimov, B. Wang, C. Han, Y. He, Q. Yuan. Brain-Specific Lipoprotein Receptors Interact with Astrocyte Derived Apolipoprotein and Mediate Neuron-Glia Lipid Shuttling. Nature Communications, 2021, 12 (1), 2408. DOI: https://doi.org/10.1038/s41467-021-22751-7.
A. Lampel, S.A. McPhee, S. Kassem, D. Sementa, T. Massarano, J. M. Aramini, Y. He, and R. V. Ulijn, Melanin-Inspired Chromophoric Microparticles Composed of Polymeric Peptide Pigments, Angew Chem Int Ed Engl, 2021, 60, 7564-7569.

2020

K. Veerasammy, Y. X. Chen, S. Sauma, M. Pruvost, D. K. Dansu, T. Choetso, T. Zhong, D. Marechal, P. Casaccia, R. Abzalimov, Y. He. Sample Preparation for Metabolic Profiling Using MALDI Mass Spectrometry Imaging. JoVE, 2020, No. 166, 62008. DOI: https://doi.org/10.3791/62008.
E. S. Duncan, A. D. Shereen, T. Gentimis, S. L. Small. Identifiable Patterns of Trait, State, and Experience in Chronic Stroke Recovery. Neurorehabil Neural Repair, 2021, 35 (2), 158–168. DOI: https://doi.org/10.1177/1545968320981953.
J. Liu, E. Likhtik, A. D. Shereen, T. A. Dennis-Tiwary, P. Casaccia. White Matter Plasticity in Anxiety: Disruption of Neural Network Synchronization During Threat-Safety Discrimination. Front Cell Neurosci, 2020, 14, 587053. DOI: https://doi.org/10.3389/fncel.2020.587053.
P. Ghobadi-Azbari, A. Jamil, F. Yavari, Z. Esmaeilpour, N. Malmir, R. Mahdavifar-Khayati, G. Soleimani, Y.-H. Cha, A. D. Shereen, M. A. Nitsche, M. Bikson, H. Ekhtiari. FMRI and Transcranial Electrical Stimulation (TES): A Systematic Review of Parameter Space and Outcomes. Progress in Neuro-Psychopharmacology and Biological Psychiatry, 2020, 110149. DOI: https://doi.org/10.1016/j.pnpbp.2020.110149.
D. K. Dansu, S. Sauma, P. Casaccia. Oligodendrocyte Progenitors as Environmental Biosensors. Semin Cell Dev Biol, 2020. DOI: https://doi.org/10.1016/j.semcdb.2020.09.012.
A. Badimon, H. J. Strasburger, P. Ayata, X. Chen, A. Nair, A. Ikegami, P. Hwang, A. T. Chan, S. M. Graves, J. O. Uweru, C. Ledderose, M. G. Kutlu, M. A. Wheeler, A. Kahan, M. Ishikawa, Y.-C. Wang, Y.-H. E. Loh, J. X. Jiang, D. J. Surmeier, S. C. Robson, et al. Negative Feedback Control of Neuronal Activity by Microglia. Nature, 2020, 586 (7829), 417–423. DOI: https://doi.org/10.1038/s41586-020-2777-8.
M. Amatruda, M. Petracca, M. Wentling, B. Inbar, K. Castro, E. Y. Chen, M. A. Kiebish, K. Edwards, M. Inglese, P. Casaccia. Retrospective Unbiased Plasma Lipidomic of Progressive Multiple Sclerosis Patients-Identifies Lipids Discriminating Those with Faster Clinical Deterioration. Scientific Reports, 2020, 10 (1), 15644. DOI: https://doi.org/10.1038/s41598-020-72654-8.
F. Li, A. Sami, H. N. Noristani, K. Slattery, J. Qiu, T. Groves, S. Wang, K. Veerasammy, Y. X. Chen, J. Morales, P. Haynes, A. Sehgal, Y. He, S. Li, Y. Song. Glial Metabolic Rewiring Promotes Axon Regeneration and Functional Recovery in the Central Nervous System. Cell Metabolism, 2020, 0 (0). DOI: https://doi.org/10.1016/j.cmet.2020.08.015.
Qiu J, Singh P, Pan G, de Paolis A, Champagne FA, Liu J, Cardoso L, Rodríguez-Contreras A. Robust range of auditory periphery development, eye opening, and brain gene expression in Wistar rat pups that experience variation in maternal backgrounds. PLoS One, 2020; 15(8):e0237933 doi: 10.1371/journal.pone.0237933
Jeff A Beeler, Devry Mourra, Roseanna M Zanca, Abigail Kalmbach, Celia Gellman, Benjamin Y Klein, Rebecca Ravenelle, Peter Serrano, Holly Moore, Stephen Rayport, Susana Mingote, Nesha S Burghardt, Vulnerable and resilient phenotypes in a mouse model of anorexia nervosa. Biological Psychiatry 2020
The iMSMS Consortium. Household Paired Design Reduces Variance and Increases Power in Multi-City Gut Microbiome Study in Multiple Sclerosis. Mult Scler, 2020, 1352458520924594. DOI: https://doi.org/10.1177/1352458520924594.
S. L. Lim, C. J. Rodriguez-Ortiz, H.-W. Hsu, J. Wu, J. Zumkehr, J. Kilian, J. Vidal, P. Ayata, M. Kitazawa. Chronic Copper Exposure Directs Microglia towards Degenerative Expression Signatures in Wild-Type and J20 Mouse Model of Alzheimer’s Disease. Journal of Trace Elements in Medicine and Biology, 2020, 62, 126578. DOI: https://doi.org/10.1016/j.jtemb.2020.126578.
P. Casaccia, E. W. G. M. Boddeke. Foreword. Glia, 2020, 68 (8), 1551–1553. DOI: https://doi.org/10.1002/glia.23841.
M. P. Fernandez, H. L. Pettibone, J. T. Bogart, C. J. Roell, C. E. Davey, A. Pranevicius, K. V. Huynh, S. M. Lennox, B. S. Kostadinov, O. T. Shafer. Sites of Circadian Clock Neuron Plasticity Mediate Sensory Integration and Entrainment. Current Biology, 2020, 0 (0). DOI: https://doi.org/10.1016/j.cub.2020.04.025.
G. M. Dmochowski, A. D. Shereen, D. Berisha, J. P. Dmochowski. Near-Infrared Light Increases Functional Connectivity with a Non-Thermal Mechanism. Cerebral Cortex Communications, 2020, 1 (1), tgaa004. DOI: https://doi.org/10.1093/texcom/tgaa004.
Samudyata, G. Castelo‐Branco, J. Liu. Epigenetic Regulation of Oligodendrocyte Differentiation: From Development to Demyelinating Disorders. Glia, 2020, glia.23820. DOI: https://doi.org/10.1002/glia.23820.
S. Sauma, P. Casaccia. Gut-Brain Communication in Demyelinating Disorders. Current Opinion in Neurobiology, 2020, 62, 92–101. DOI: https://doi.org/10.1016/j.conb.2020.01.005.
C. Yattah, M. Hernandez, D. Huang, H. Park, W. Liao, P. Casaccia. Dynamic Lamin B1-Gene Association During Oligodendrocyte Progenitor Differentiation. Neurochem Res, 2020. DOI: https://doi.org/10.1007/s11064-019-02941-y.
P. Ayata, A. Schaefer. Innate Sensing of Mechanical Properties of Brain Tissue by Microglia. Current Opinion in Immunology, 2020, 62, 123–130. DOI: https://doi.org/10.1016/j.coi.2020.01.003.
P. Shrestha, P. Ayata*, P. Herrero-Vidal, F. Longo, A. Gastone, J. E. LeDoux, N. Heintz, E. Klann. Cell-Type-Specific Drug-Inducible Protein Synthesis Inhibition Demonstrates That Memory Consolidation Requires Rapid Neuronal Translation. Nat Neurosci, 2020, 23 (2), 281–292. DOI: https://doi.org/10.1038/s41593-019-0568-z. (* co-first author)
P. Casaccia. Emerging Concepts in Neuroscience Research: 2019 Highlights. The Lancet Neurology, 2020, 19 (1), 21–22. DOI: https://doi.org/10.1016/S1474-4422(19)30452-1.

2019

Z. Esmaeilpour, A. D. Shereen, P. Ghobadi‐Azbari, A. Datta, A. J. Woods, M. Ironside, J. O’Shea, U. Kirk, M. Bikson, H. Ekhtiari. Methodology for TDCS Integration with FMRI. Hum Brain Mapp, 2020, 41 (7), 1950–1967. DOI: https://doi.org/10.1002/hbm.24908.

B. J. Piggott, C. J. Peters, Y. He, X. Huang, S. Younger, L. Y. Jan, Y. N. Jan. Paralytic, the Drosophila Voltage-Gated Sodium Channel, Regulates Proliferation of Neural Progenitors. Genes Dev., 2019, 33 (23–24), 1739–1750. DOI: https://doi.org/10.1101/gad.330597.119.

S. Sauma, P. Casaccia. Does the Gut Microbiota Contribute to the Oligodendrocyte Progenitor Niche? Neuroscience Letters, 2019, 134574. DOI: https://doi.org/10.1016/j.neulet.2019.134574.

Mingote S*, Amsellem A, Kempf A, Rayport S, Chuhma N. Dopamine-Glutamate Neuron Projections to the Nucleus Accumbens Medial Shell and Behavioral Switching. In special issue: The physiology and pathophysiology of basal ganglia. Neurochemistry International, 2019; 129: 104482. Review. doi: 10.1016/j.neuint.2019.104482

Lauren Kosten, Golam MI Chowdhury, Susana Mingote, Steven Staelens, Douglas L Rothman, Kevin L Behar, Stephen Rayport, Glutaminase activity in GLS1 Het mouse brain compared to putative pharmacological inhibition by ebselen using ex vivo MRS. Neurochemistry international vol. 129, p. 104508

M. Wentling, C. Lopez-Gomez, H.J. Park, M. Amatruda, A. Ntranos, J. Aramini, M. Petracca, T. Rusielewicz, E. Chen, V. Tolstikov, M. Kiebish, V. Fossati, M. Inglese, C.M. Quinzii, I. Katz Sand, P. Casaccia. A metabolic perspective on CSF-mediated neurodegeneration in multiple sclerosis. Brain, 2019. DOI: 10.1093/brain/awz201 (press release)

V. Bonnefil, K. Dietz, M. Amatruda, M. Wentling, A.V. Aubry, J.L. Dupree, G. Temple, H.J. Park, N.S. Burghardt, P. Casaccia, J. Liu. Region-specific myelin differences define behavioral consequences of chronic social defeat stress in mice. ELife, 2019, 8. DOI: 10.7554/eLife.40855.001. Podcast: https://elifesciences.org/podcast/episode60#933.

L.L. Boshans, D.C. Factor, V. Singh, J. Liu, C. Zhao, I. Mandoiu, Q.R. Lu, P. Casaccia, P.J. Tesar, A. Nishiyama. The Chromatin Environment Around Interneuron Genes in Oligodendrocyte Precursor Cells and Their Potential for Interneuron Reprograming. Frontiers in Neuroscience, 2019, 13. DOI: 10.3389/fnins.2019.00829

P. Shrestha, P. Ayata*, P. Herrero-Vidal, A. Gastone, JE LeDoux, N. Heintz, E. Klann. Chemogenetic evidence that rapid neuronal de novo protein synthesis is required for consolidation of long-term memory. bioRxiv. 2019 704965 (* co-first author)

K. Castro, A. Ntranos, M. Amatruda, M. Petracca, P. Kosa, E.Y. Chen, J. Morstein, D. Trauner, C.T. Watson, M.A. Kiebish, B. Bielekova, M. Inglese, I. Katz Sand, P. Casaccia. Body Mass Index in Multiple Sclerosis modulates ceramide-induced DNA methylation and disease course. EBioMedicine, 2019. DOI: 10.1016/j.ebiom.2019.03.087 (press release)

A.M. Falcão, M. Meijer, A. Scaglione, P. Rinwa, E. Agirre, J. Liang, S.C. Larsen, A. Heskol, R. Frawley, M. Klingener, M. Varas-Godoy, A.A.S.F. Raposo, P. Ernfors, D.S. Castro, M.L. Nielsen, P. Casaccia, G. Castelo-Branco. PAD2-Mediated Citrullination Contributes to Efficient Oligodendrocyte Differentiation and Myelination. Cell Reports, 2019, 4. DOI: 10.1016/j.celrep.2019.03.108

Tsai, P. Casaccia. Mechano-Modulation of Nuclear Events Regulating Oligodendrocyte Progenitor Gene Expression. Glia, 2019, 67 (7), 1229–1239. DOI: https://doi.org/10.1002/glia.23595.

A. Ntranos, V. Ntranos, V. Bonnefil, J. Liu, S. Kim-Schulze, Y. He, Y. Zhu, R. Brandstadter, C.T. Watson, A.J. Sharp, I. Katz Sand, P. Casaccia. Fumarates target the metabolic-epigenetic interplay of brain-homing T cells in multiple sclerosis. Brain, 2019, 142(3), 647-661. DOI: 10.1093/brain/awy344

2018

J. Liu, K. Dietz, G.E. Hodes, S.J. Russo, P. Casaccia. Widespread transcriptional alternations in oligodendrocytes in the adult mouse brain following chronic stress. Developmental neurobiology, 2018 Feb;78(2):152-162.. DOI: doi.org/10.1002/dneu.22533

R.W. Motl, E.M. Mowry, D.M. Ehde, N.G. LaRocca, K.E. Smith, K. Costello, L. Shinto, A.V. Ng, A.B. Sullivan, B. Giesser, K.K. McCully, B. Fernhall, M. Bishop, M. Plow, P. Casaccia, N.D. Chiaravalloti. Wellness and multiple sclerosis: The National MS Society establishes a Wellness Research Working Group and research priorities. Multiple sclerosis (Houndmills, Basingstoke, England), 2018 Mar;24(3):262-267.. DOI: doi.org/10.1177/1352458516687404

A. Pathak, E.M. Stanley, F.E. Hickman, N. Wallace, B. Brewer, D. Li, S. Gluska, E. Perlson, S. Fuhrmann, K. Akassoglou, F. Bronfman, P. Casaccia, D.T. Burnette, B.D. Carter. Retrograde Degenerative Signaling Mediated by the p75 Neurotrophin Receptor Requires p150Glued Deacetylation by Axonal HDAC1. Developmental cell, 2018 Aug 6;46(3):376-387.e7.. DOI: doi.org/10.1016/j.devcel.2018.07.001

I. K. Sand, Y. Zhu, A. Ntranos, J.C. Clemente, E. Cekanaviciute, R. Brandstadter, E. Crabtree-Hartman, S. Singh, Y. Bencosme, J. Debelius, R. Knight, B.A.C. Cree, S.E. Baranzini, P. Casaccia. Disease-modifying therapies alter gut microbial composition in MS. Neurology – Neuroimmunology Neuroinflammation, 2018, 6, 1. DOI: 10.1212/nxi.0000000000000517

A. Scaglione, J. Patzig, J. Liang, R. Frawley, J. Bok, A. Mela, C. Yattah, J. Zhang, S.X. Teo, T. Zhou, S. Chen, E. Bernstein, P. Canoll, E. Guccione, P. Casaccia. PRMT5-mediated regulation of developmental myelination. Nat Commun, 2018. DOI: 10.1038/s41467-018-04863-9.

A.T. McKenzie, M. Wang, M.E. Hauberg, J.F. Fullard, A. Kozlenkov, A. Keenan, Y.L. Hurd, S. Dracheva, P. Casaccia, P. Roussos, B. Zhang. Brain Cell Type Specific Gene Expression and Co-expression Network Architectures. Sci Rep, 2018. DOI: 10.1038/s41598-018-27293-5.

A. Schlüter, J. Sandoval, S. Fourcade, A. Díaz-Lagares, M. Ruiz, P. Casaccia, M. Esteller, A. Pujol. Epigenomic signature of adrenoleukodystrophy predicts compromised oligodendrocyte differentiation. Brain Pathol, 2018. DOI: 10.1111/bpa.12595.

P. Casaccia, G. Corfas. Introduction to the special issue on myelin plasticity in the central nervous system. Dev Neurobiol, 2018. DOI: 10.1002/dneu.22575.

K. Castro, P. Casaccia. Epigenetic modifications in brain and immune cells of multiple sclerosis patients. Mult Scler, 2018. DOI: 10.1177/1352458517737389.

A. Ntranos, P. Casaccia. The Microbiome-Gut-Behavior Axis: Crosstalk Between the Gut Microbiome and Oligodendrocytes Modulates Behavioral Responses. Neurotherapeutics. 2018 Jan, 15 ,1, 31-35. DOI: 10.1007/s13311-017-0597-9

E. Cekanaviciute, A. Pröbstel, A. Thomann, T.F. Runia, P. Casaccia, I.K. Sand, E. Crabtree, S. Singh, J. Morrissey, P. Barba, R. Gomez, R. Knight, S. Mazmanian, J. Graves, B.A.C. Cree, S.S. Zamvil, S.E. Baranzini. Multiple Sclerosis-Associated Changes in the Composition and Immune Functions of Spore-Forming Bacteria. mSystems, 2018 Nov 6;3(6):e00083-18.. DOI: doi.org/10.1128/msystems.00083-18

V. Aubry, H. Khandaker, R. Ravenelle, I. S. Grunfeld, V. Bonnefil, K. L. Chan, F. Cathomas, J. Liu, G. E. Schafe, N. S. Burghardt. A Diet Enriched with Curcumin Promotes Resilience to Chronic Social Defeat Stress. Neuropsychopharmacol, 2019, 44 (4), 733–742. DOI: https://doi.org/10.1038/s41386-018-0295-2.