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The effect of poly I:c or LPS priming on the therapeutic efficacy of mesenchymal stem cells in an adjuvant-induced arthritis rat model

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PHARMAcOLOGIcAL REPORTS 2022年第4期74卷 654-668页

作者： Zolfaghari, Samira Milan, Peiman Brouki Dehpour, Ahmad Reza Fomeshi, Motahareh Rajabi Eskandari, Fatemeh Ebrahimi, Loghman Hashemi, Seyed Mahmoud Joghataei, Mohammad Taghi Iran Univ Med Sci Cellular & Mol Res Ctr Tehran *** Iran Iran Univ Med Sci Fac Adv Technol Med Dept Tissue Engn & Regenerat Med Tehran Iran Univ Tehran Med Sci Sch Med Dept Pharmacol Tehran Iran Shahid Beheshti Univ Med Sci Sch Med Dept Immunol Tehran *** Iran

Background The immunomodulatory properties of mesenchymal stem cells (MScs) have made them a prospective treatment option for inflammatory and autoimmune disorders. Recent studies have found an association between the immunomodulatory function of MScs and Toll-like receptors (TLRs). Here, we investigated the effect of priming with lipopolysaccharide (LPS) as TLR4 ligand or polyinosinic:polycytidylic acid (poly I:c) as TLR3 ligand on the immunomodulatory function of adipose-derived MScs (ADMScs) in vitro and for the first time in an adjuvant-induced arthritis model (AIA). Methods ADMScs were treated with LPS or poly I:c for 1 h. Splenocyte proliferation in the presence of primed ADMScs was assessed in vitro using an MTT assay. Next, we investigated the therapeutic effect of primed ADMScs in vivo. Male Wistar rats were infused with complete Freund's adjuvant (cFA) to develop arthritis and then intraperitoneally treated with not-primed, poly I:c- or LPS-primed ADMScs. clinical signs, histopathological alteration, and serum and spleen cytokine levels were analyzed. Results Poly I:c-primed ADMScs significantly reduced splenocytes proliferation, while ADMScs primed with LPS increased splenocytes proliferation. Furthermore, poly I:c-primed ADMScs significantly alleviated the clinical and histopathological severity and the secretion of inflammatory cytokines associated with Th17/Th1 such as IL-17 and IFN-gamma. Poly I:c-primed ADMScs also increased cytokines IL-10 and TGF-beta. TNF-alpha and IL-6 Levels were also markedly diminished in the serum of AIA animals treated with poly I:c-primed ADMScs. In contrast, priming ADMScs with LPS significantly reduced the therapeutic effect of ADMScs in AIA animals. conclusion As a result of these findings, poly I:c priming may be a new technique for improving the therapeutic effects of MScs in arthritic disorders.

关键词： Mesenchymal stem cells Toll-like receptor Poly I c LPS Adjuvant-induced arthritis Immunomodulation

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An assessment of the spontaneous locomotor activity of BALB/c mice

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JOURNAL OF PHARMAcOLOGIcAL ScIENcES 2022年第2期149卷 46-52页

作者： Miyazaki, Yusuke Kobayashi, Koji Matsushita, Seiji Shimizu, Naoyuki Murata, Takahisa Univ Tokyo Grad Sch Agr & Life Sci Dept Anim Radiol Tokyo Japan

The surrounding environment affects the behavior of an animal. Therefore, long-term observation of an animal's behavior in its natural breeding environment is an effective approach to predict and understand animal behavior in greater detail. Spontaneous locomotor activity (SLA), the movement of animals in their breeding environment, is one of the most important behavioral indices for experimental animals. We here established an SLA measurement system using image analyses to obtain basic data from BALB/c mice. To record the movement of the mice, we used an infrared video camera. SLA of mice were calculated by detecting their geometric center in each frame. This system could detect the mouse correctly more than 99.999% in all frames. Further, we investigated the effects of habituation, age, and sex on the SLA of BALB/c mice. Three days of habituation were required to decrease the SLA of mice placed in novel cages. The 16- and 32-week-old mice were less active than 4-week-old mice. No significant differences were detected between males and females. We also found that BALB/c and c57BL/6 mice differed in their active and resting rhythms. In conclusion, we developed an SLA measurement system and obtained basic SLA data from BALB/c mice. (c) 2022 The Authors. Production and hosting by Elsevier B.V. on behalf of Japanese Pharmacological Society.

关键词： Spontaneous locomotor activity Behavior BALB c Experimental animal Video analysis

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Functional analysis of a novel de novo variant in PPP5c associated with microcephaly, seizures, and developmental delay

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MOLEcULAR GENETIcS AND METABOLISM 2022年第1期136卷 65-73页

作者： Fielder, Sara M. Rosenfeld, Jill A. Burrage, Lindsay c. Emrick, Lisa Lalani, Seema Attali, Ruben Bembenek, Joshua N. Hieu Hoang Baldridge, Dustin Silverman, Gary A. Schedl, Tim Pak, Stephen c. Washington Univ Dept Pediat Sch Med St Louis MO 63110 USA Baylor Coll Med Dept Mol & Human Genet Houston TX 77030 USA Texas Childrens Hosp Houston TX 77030 USA Emedgene Technol Genom Res Dept IL-6744332 Tel Aviv Israel Wayne State Univ CS Mott Ctr Human Growth & Dev Dept Obstet & Gynecol Sch Med Detroit MI 48201 USA Washington Univ Dept Genet Sch Med St Louis MO 63110 USA

We describe a proband evaluated through the Undiagnosed Diseases Network (UDN) who presented with microcephaly, developmental delay, and refractory epilepsy with a de novo ***47Thr missense variant in the protein phosphatase gene, PPP5c. This gene has not previously been associated with a Mendelian disease, and based on the population database, gnomAD, the gene has a low tolerance for loss-of-function variants (pLI = 1, o/e = 0.07). We functionally evaluated the PPP5c variant in c. elegans by knocking the variant into the orthologous gene, pph-5, at the corresponding residue, Ala48Thr. We employed assays in three different biological processes where pph-5 was known to function through opposing the activity of genes, mec-15 and sep-1. We demonstrated that, in contrast to control animals, the pph-5 Ala48Thr variant suppresses the neurite growth phenotype and the GABA signaling defects of mec-15 mutants, and the embryonic lethality of sep-1 mutants. The Ala48Thr variant did not display dominance and behaved similarly to the reference pph-5 null, indicating that the variant is likely a strong hypomorph or complete loss-of-function. We conclude that pph-5 Ala48Thr is damaging in c. elegans. By extension in the proband, PPP5c ***47Thr is likely damaging, the de novo dominant presentation is consistent with haplo-insufficiency, and the PPP5c variant is likely responsible for one or more of the proband's phenotypes.(c) 2022 The Authors. Published by Elsevier Inc. This is an open access article under the cc BY-Nc-ND license (http:// ***/licenses/by-nc-nd/4.0/).

关键词： PPP5c PPH-5 Microcephaly Developmental delay c elegans

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M05B5.4 (lysosomal phospholipase A2) promotes disintegration of autophagic vesicles to maintain c. elegans development

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AUTOPHAGY 2022年第3期18卷 595-607页

作者： Li, Yuan Wang, Xin Li, Meijiao Yang, chonglin Wang, Xiaochen Chinese Acad Sci Inst Biophys CAS Ctr Excellence Biomacromol Natl Lab Biomacromol Beijing 100101 Peoples R China Yunnan Univ Sch Life Sci State Key Lab Conservat & Utilizat Bioresources Y Kunming Yunnan Peoples R China Yunnan Univ Sch Life Sci Ctr Life Sci Kunming Yunnan Peoples R China Univ Chinese Acad Sci Coll Life Sci Beijing Peoples R China

The autophagosome has two lipid bilayer membranes. The outer membrane fuses with the lysosome, while the inner membrane is degraded to release autophagic contents for degradation. It remains unclear how the inner vesicle of the autophagosome (called the autophagic vesicle) is disintegrated after autophagosome-lysosome fusion. Here, we identified c. elegans LPLA-2/M05B5.4 as a key enzyme that degrades membranous material in lysosomes. LPLA-2 is homologous to human PLA2G15, a lysosomal phospholipase A2 family protein that catalyzes cleavage of membrane phospholipids. We found that loss of LPLA-2 causes accumulation of large membrane whor ls in enlarged lysosomes and both phenotypes are suppressed by blocking macroautophagy/autophagy. Moreover, autophagic vesicles persisted in enlarged lysosomes in PLA2G15 knockdown cells and lpla-2(lf) mutants, which suggests that the breakdown of the inner autophagosomal membrane in lysosomes is impaired. lpla-2(lf) mutants exhibit severe defects in both embryonic and larval development. Our data suggest that disintegration of the inner autophagosomal membrane by LPLA-2 promotes the release and subsequent degradation of autophagic contents in lysosomes, which is essential for c. elegans development.

关键词： Autophagic vesicle c elegans development inner autophagosomal membrane lysosomal phospholipase A2 lysosome

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Long-lasting housing environment manipulation and acute loss of environmental enrichment impact BALB/c mice behaviour in multiple functional domains

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EUROPEAN JOURNAL OF NEUROScIENcE 2022年第5期55卷 1118-1140页

作者： Sukegawa, Momoe Yoshihara, Toru Hou, Shengqun Asano, Masahide Hannan, Anthony J. Wang, Dan Ohtan RIKEN Ctr Biosyst Dynam Res BDR Kobe Hyogo Japan Kyoto Univ Grad Sch Biostudies Kyoto Japan Kyoto Univ Inst Integrated Cell Mat Sci iCeMS Kyoto Japan Kyoto Univ Grad Sch Med Inst Lab Anim Kyoto Japan Univ Melbourne Dept Anat & Neurosci Parkville Vic Australia Melbourne Brain Ctr Parkville Vic Australia

Understanding environmental influences on individuals' behaviour is challenging. Here we have investigated the housing impact of 9 weeks of enriched environment (EE) and social isolation (SI) and the impact of abrupt deprivation of EE (enrichment removal: ER) on BALB/c mice. compared with the widely used c57BL/6 strain in research, BALB/c synthesises serotonin less efficiently due to a genetic variation and thus may potentially represent human populations at higher risk of stress-related disorders. We assessed the effects of EE and SI by conducting a behavioural test battery and the effects of acute ER by monitoring homecage activities and social behaviour. We found that EE and SI impact BALB/c's physiological states and behavioural performances from lower to higher cognitive processes: increased body weight, increased rectal temperature, altered performance in motor and sensory tasks, the activity level in a novel environment and altered performance in tests of anxiety-like behaviour, stress-coping strategies and learning and memory. Furthermore, acute ER triggered stress/frustration-like behaviour in BALB/c, with increased aggression, increased social distancing and disrupted daily/nightly activities. Our results demonstrate that long-lasting housing manipulation such as EE and SI, impact behaviour via multilayered processes over a wide range of functional domains, and unforeseen change to a negative environment, ER, is a major stressor that causes behavioural and psychological consequences through environment-gene interactions, a model of direct relevance to human health.

关键词： BALB c behavioural test battery enriched environment enrichment removal social isolation

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Using a GIS-based spatial approach to determine the optimal locations of bikeshare stations: The case of Washington D.c

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TRANSPORT POLIcY 2022年 127卷 48-60页

作者： Ebrahimi, Zhila Dehdari Momenitabar, Mohsen Nasri, Arefeh A. Mattson, Jeremy North Dakota State Univ Coll Business Dept Transportat Logist & Finance Fargo ND 58108 USA Univ Maryland Sch Architecture Planning & Preservat College Pk MD 20742 USA

Today, the number of cities implementing bike-share programs is remarkably increasing. One of the critical elements of implementing a successful bike-share program is integrating it with other transportation modes such as bus and metro and extending its coverage in high-density residential and employment areas to encourage more demand. In this study, we utilize a GIS-based method to visualize the spatial distribution of bike-share stations using the location-allocation problem, using the capital bike-share program in Washington D.c. metropolitan areas as a study area. We chose Washington D.c. as it was one of the first cities in the United States that launched a bike-share program and currently has one of the largest bike-share systems across the country. The location -allocation problem, including Target Market Share (TMS) and Maximize coverage and Minimize Facility (McMF), is considered to analyze the accessibility of promoting transit modes with bike-share systems across the District of columbia. Location-allocation models are investigated to determine the potential bike station loca-tions accessible to the maximum population and within a 300-m buffer around public transit stations (e.g., bus, metro). The results show that the bike-share system in Washington D.c. is more accessible for transit users as an access/egress mode. At the same time, in areas farther away from downtown D.c., docking stations are more distanced apart and offer less coverage, especially in residential-only areas. Finally, our methodology can potentially be utilized for optimal station location allocation in any other city where maximum exposure is required.

关键词： Bike-share program Public transit Location-allocation model GIS Washington D c

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PolyI:c attenuates transforming growth factor-beta signaling to induce cytostasis of surrounding cells by secreted factors in triple-negative breast cancer

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cANcER ScIENcE 2022年第3期113卷 940-949页

作者： Tamura, Yusuke Tsutsumi, Shuichi Miyazono, Kohei Koinuma, Daizo Univ Tokyo Grad Sch Med Dept Mol Pathol 7-3-1 Bunkyo Ku Tokyo 1130033 Japan Univ Tokyo Res Ctr Adv Sci & Technol Genome Sci Div Tokyo Japan

The activation of RIG-I-like receptor (RLR) signaling in cancer cells is widely recognized as a critical cancer therapy method. The expected mechanism of RLR ligand-mediated cancer therapy involves the promotion of cancer cell death and strong induction of interferon (IFN)-beta that affects the tumor microenvironment. We have recently shown that activation of RLR signaling in triple-negative breast cancer cells (TNBc) attenuates transforming growth factor-beta (TGF-beta) signaling, which partly contributes to the promotion of cancer cell pyroptosis. However, the consequences of suppression of TGF-beta signaling by RLR ligands with respect to IFN-beta-mediated tumor suppression are not well characterized. This study showed that transfection of a typical RLR ligand polyI:c in cancer cells produces significant levels of IFN-beta, which inhibits the growth of the surrounding cancer cells. In addition, IFN-beta-induced cell cycle arrest in surrounding cancer cells was inhibited by the expression of constitutively active Smad3. constitutively active Smad3 suppresses IFN-beta expression through the alleviation of IFN regulatory factor 3 binding to the canonical target genes, as suggested by chIP sequencing analysis. Based on these findings, a new facet of the protumorigenic function of TGF-beta that suppresses IFN-beta expression is suggested when RLR-mediated cancer treatment is used in TNBc.

关键词： IFN-beta IRF3 polyI c TGF-beta TNBc

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Pre- and post-conditioning with poly I:c exerts neuroprotective effect against cerebral ischemia injury in animal models: A systematic review and meta-analysis

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cNS NEUROScIENcE & THERAPEUTIcS 2022年第8期28卷 1168-1182页

作者： Khan, Zeeshan Ahmad Sumsuzzman, Dewan Md choi, Jeonghyun Kamenos, George Hong, Yonggeun Coll Healthcare Med Sci & Engn Dept Phys Therapy 197 Inje Ro Gimhae 50834 Gyeongsangnam D South Korea Inje Univ Biohlth Prod Res Ctr BPRC Gimhae South Korea Inje Univ Res Ctr Aged Life Redesign RCAR Gimhae South Korea Inje Univ Dept Rehabil Sci Grad Sch Gimhae South Korea

Background Toll-like receptor (TLR) agonist polyinosinic-polycytidylic acid (poly I:c) exerts neuroprotective effects against cerebral ischemia (cI), but concrete evidence supporting its exact mechanism of action is unclear. Methods We evaluated the neuroprotective role of poly I:c by assessing cI indicators such as brain infarct volume (BIV), neurological deficit score (N.S.), and signaling pathway proteins. Moreover, we performed a narrative review to illustrate the mechanism of action of TLRs and their role in cI. Our search identified 164 articles and 10 met the inclusion criterion. Results Poly I:c reduces BIV and N.S. (p = 0.00 and p = 0.03). Interestingly, both pre- and post-conditioning decrease BIV (prec p = 0.04 and postc p = 0.00) and N.S. (prec p = 0.03 and postc p = 0.00). Furthermore, poly I:c upregulates TLR3 [SMD = 0.64;cIs (0.56, 0.72);p = 0.00], downregulates nuclear factor-kappa B (NF-kappa B) [SMD = -1.78;cIs (-2.67, -0.88);p = 0.0)], and tumor necrosis factor alpha (TNF-alpha) [SMD = -16.83;cIs (-22.63, -11.02);p = 0.00]. conclusion We showed that poly I:c is neuroprotective and acts via the TLR3/NF-kappa B/TNF-alpha pathway. Our review indicated that suppressing TLR 2/4 may illicit neuroprotection against cI. Further research on simultaneous activation of TLR3 with poly I:c and suppression of TLR 2/4 might open new vistas for the development of therapeutics against cI.

关键词： cerebral ischemia meta-analysis poly I c systematic review toll-like receptors

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A practical approach for the porting of the Ravenscar profile from ADA to c: method, rules adaptation and supporting tools

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INTERNATIONAL JOURNAL OF cOMPUTER APPLIcATIONS IN TEcHNOLOGY 2022年第2期70卷 85-93页

作者： Rinaldi, claudia Romano, Fabio Serri, Paolo Tiberi, Luca Univ Aquila Ctr Excellence DEWS Laquila Italy Thales Alenia Space Italia TASI Laquila Italy

While conceiving with hard real time systems, determinism is the main requirement that must be satisfied in order to properly predict their behaviour as required by their definition. For achieving this purpose an available solution is restricting Ada language tasking features to the Ada Ravenscar profile subset. This paper presents a solution to apply the Ravenscar profile concepts in systems where the tasking management is based on RTEMS real-time operating system, and c and Ada languages are used together. Moreover, a SW tool to automatically check the compliance with Ravenscar is proposed and the outcomes of some experimental activities proving the effectiveness of the SW tool are discussed.

关键词： Ravenscar profile Ada c RTEMS

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GLH-1/Vasa represses neuropeptide expression and drives spermiogenesis in the c. elegans germline

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DEVELOPMENTAL BIOLOGY 2022年第0期492卷 200-211页

作者： Rochester, Jesse D. Min, Hyemin Gajjar, Gita A. Sharp, catherine S. Maki, Nathaniel J. Rollins, Jarod A. Keiper, Brett D. Graber, Joel H. Updike, Dustin L. Kathryn W Davis Ctr Regenerat Biol & Aging Mt Desert Isl Biol Lab Bar Harbor ME 04609 USA Univ Maine Grad Sch Biomed Sci & Engn Orono ME USA East Carolina Univ Brody Sch Med Dept Biochem & Mol Biol Greenville NC USA

Germ granules harbor processes that maintain germline integrity and germline stem cell capacity. Depleting core germ granule components in c. elegans leads to the reprogramming of germ cells, causing them to express markers of somatic differentiation in day-two adults. Somatic reprogramming is associated with complete sterility at this stage. The resulting germ cell atrophy and other pleiotropic defects complicate our understanding of the initiation of reprogramming and how processes within germ granules safeguard the totipotency and immortal potential of germline stem cells. To better understand the initial events of somatic reprogramming, we examined total mRNA (transcriptome) and polysome-associated mRNA (translatome) changes in a precision full-length deletion of glh-1, which encodes a homolog of the germline-specific Vasa/DDX4 DEAD-box RNA helicase. Fertile animals at a permissive temperature were analyzed as young adults, a stage that precedes by 24 h the previously determined onset of somatic reporter-gene expression in the germline. Two significant changes are observed at this early stage. First, the majority of neuropeptide-encoding transcripts increase in both the total and polysomal mRNA fractions, suggesting that GLH-1 or its effectors suppress this expression. Second, there is a significant decrease in Major Sperm Protein (MSP)-domain mRNAs when glh-1 is deleted. We find that the presence of GLH-1 helps repress spermatogenic expression during oogenesis, but boosts MSP expression to drive spermiogenesis and sperm motility. These insights define an early role for GLH-1 in repressing somatic reprogramming to maintain germline integrity.

关键词： GLH-1 Vasa DDX4 Germ granules P granules c elegans Germline Sperm Spermiogenesis Major sperm protein MSP Neuropeptides Polysomes

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