Synergistic Combination of Dual Clays in Multilayered Nanocomposites for Enhanced Flame Retardant Properties.

In an effort to reduce the flammability of synthetic polymeric materials such as cotton fabrics and polyurethane foam (PUF), hybrid nanocoatings are prepared by layer-by-layer assembly. Multilayered nanocomposites of a cationic polyelectrolyte, poly(diallyldimethylammonium chloride) (PDDA), are paired with two kinds of clay nanoplatelets, montmorillonite (MMT) and vermiculite (VMT). The physical properties such as thickness and mass and thermal behaviors in clay-based nanocoatings with and without incorporation of tris buffer are compared to assess the effectiveness of amine salts on flame retardant (FR) performances. A PDDA-tris/VMT-MMT system, in which tris buffer is introduced into the cationic PDDA aqueous solution, produces a thicker and heavier coating. Three different systems, including PDDA/MMT, PDDA/VMT-MMT, and PDDA-tris/VMT-MMT, result in conformal coating, retaining the weave structure of the fabrics after being exposed to a vertical and horizontal flame test, while the uncoated sample is completely burned out. The synergistic effects of dual clay-based hybrid nanocoatings are greatly improved by adding amine salts. Cone calorimetry reveals that the PDDA-tris/VMT-MMT-coated PUF eliminates a second peak heat release rate and significantly reduces other FR performances, compared to those obtained from the clay-based multilayer films with no amine salts added. Ten bilayers of PDDA-tris/VMT-MMT (≈250 nm thick) maintain the shape of foam after exposure to a butane torch flame for 12 s. As for practical use of these nanocomposites in real fire disasters, spray-assisted PDDA-tris/VMT-MMT multilayers on woods exhibit high resistance over flammability. Improved fire resistance in PDDA-tris/VMT-MMT is believed to be due to the enhanced char yield through the addition of tris buffer that promotes the deposition of more clay particles while retaining a highly ordered deposition of a densely packed nanobrick wall structure. This work demonstrates the ability to impart significant fire resistance to synthetic polymer materials in a fully renewable nanocoating that uses environmentally benign chemistry.

Predisposal of Interferon Regulatory Factor 1 Deficiency to Accumulate DNA Damage and Promote Osteoarthritis Development in Cartilage.

OBJECTIVE: Interferon regulatory factor 1 (IRF1) is a transcriptional regulator conventionally associated with immunomodulation. Recent molecular analyses mapping DNA binding sites of IRF1 have suggested its potential function in DNA repair. However, the physiologic significance of this noncanonical function remains unexplored. Here, we investigated the role of IRF1 in osteoarthritis (OA), a condition marked by senescence and chronic joint inflammation. METHODS: OA progression was examined in wild-type and Irf1-/- mice using histologic assessments and microcomputed tomography analysis of whole-joint OA manifestations and behavioral assessments of joint pain. An integrated analysis of assay for transposase-accessible chromatin with sequencing and whole transcriptome data was conducted for the functional assessment of IRF1 in chondrocytes. The role of IRF1 in DNA repair and senescence was investigated by assaying γ-H2AX foci and senescence-associated beta-galactosidase activity. RESULTS: Our genome-wide investigation of IRF1 footprinting in chondrocytes revealed its primary occupancies in the promoters of DNA repair genes without noticeable footprint patterns in those of interferon-responsive genes. Chondrocytes lacking IRF1 accumulated irreversible DNA damage under oxidative stress, facilitating their entry into cellular senescence. IRF1 was down-regulated in the cartilage of human and mouse OA. Although IRF1 overexpression did not elicit an inflammatory response in joints or affect OA development, genetic deletion of Irf1 caused enhanced chondrocyte senescence and exacerbated post-traumatic OA in mice. CONCLUSION: IRF1 offers DNA damage surveillance in chondrocytes, protecting them from oxidative stress associated with OA risk factors. Our study provides a crucial and cautionary perspective that compromising IRF1 activity renders chondrocytes vulnerable to cellular senescence and promotes OA development.

Hunting for the vulnerability in chondrosarcoma by tracing metabolic and genetic links.

In this Backstory, we narrate our journey in establishing a multidisciplinary team for sarcoma research and uncovering vulnerabilities in chondrosarcoma cells associated with their NAD+ dependencies for survival.1 Our findings hold promise for exploitation, yielding a synergistic cytotoxic effect when combined with systemic therapy.

Effectiveness of mid thoracic spine mobilization on postural balance and gait ability in subacute stroke patients: A randomized clinical trial.

BACKGROUND: Although mulligan sustained natural apophyseal glides (SNAG) and maitland mobilization (MM) are common interventions for musculoskeletal disease, no study has directly compared the effectiveness of mid-thoracic spine mobilization in subacute stroke patients. OBJECTIVE: To investigate the effects of mid-thoracic spine mobilization (SNAG vs. MM) on postural balance and gait ability in subacute stroke patients. METHODS: Fifty subacute stroke patients were randomly allocated to the SNAG (n= 17), MM (n= 16), and control (n= 17) groups, each receiving a neuro-developmental therapy program for four successive weeks. The SNAG and MM groups additionally received mid-thoracic spine mobilization (T4∼8). The primary outcome measure was postural sway, and secondary outcome measures included the five times sit-to-stand test (FTSST), functional reach test (FRT), 10-m walk test (10MWT), 6-minute walk test (6MWT) and global rating of change (GRC). RESULTS: Participants reported no adverse events, and there was no loss to follow-up. The SNAG and MM group patients demonstrated significant improvements (p< 0.05) in postural sway, FTSST, FRT, 10MWT, and 6MWT compared with those in the control group, with no between-group differences. CONCLUSIONS: Mid-thoracic spine mobilization allows significant improvements in postural balance and gait ability in subacute stroke patients, with no differences between the SNAG and MM techniques.

Decoupling NAD+ metabolic dependency in chondrosarcoma by targeting the SIRT1-HIF-2α axis.

Chondrosarcomas represent the second most common primary bone malignancy. Despite the vulnerability of chondrosarcoma cells to nicotinamide adenine dinucleotide (NAD+) depletion, targeting the NAD+ synthesis pathway remains challenging due to broad implications in biological processes. Here, we establish SIRT1 as a central mediator reinforcing the dependency of chondrosarcoma cells on NAD+ metabolism via HIF-2α-mediated transcriptional reprogramming. SIRT1 knockdown abolishes aggressive phenotypes of chondrosarcomas in orthotopically transplanted tumors in mice. Chondrosarcoma cells thrive under glucose starvation by accumulating NAD+ and subsequently activating the SIRT1-HIF-2α axis. Decoupling this link via SIRT1 inhibition unleashes apoptosis and suppresses tumor progression in conjunction with chemotherapy. Unsupervised clustering analysis identifies a high-risk chondrosarcoma patient subgroup characterized by the upregulation of NAD+ biosynthesis genes. Finally, SIRT1 inhibition abolishes HIF-2α transcriptional activity and sensitizes chondrosarcoma cells to doxorubicin-induced cytotoxicity, irrespective of underlying pathways to accumulate intracellular NAD+. We provide system-level guidelines to develop therapeutic strategies for chondrosarcomas.

Sarcoma Immunotherapy: Confronting Present Hurdles and Unveiling Upcoming Opportunities.

Sarcomas are rare and heterogeneous mesenchymal neoplasms originating from the bone or soft tissues, which pose significant treatment challenges. The current standard treatment for sarcomas consists of surgical resection, often combined with chemo- and radiotherapy; however, local recurrence and metastasis remain significant concerns. Although immunotherapy has demonstrated promise in improving long-term survival rates for certain cancers, sarcomas are generally considered to be relatively less immunogenic than other tumors, presenting substantial challenges for effective immunotherapy. In this review, we examine the possible opportunities for sarcoma immunotherapy, noting cancer testis antigens expressed in sarcomas. We then cover the current status of immunotherapies in sarcomas, including progress in cancer vaccines, immune checkpoint inhibitors, and adoptive cellular therapy and their potential in combating these tumors. Furthermore, we discuss the limitations of immunotherapies in sarcomas, including a low tumor mutation burden and immunosuppressive tumor microenvironment, and explore potential strategies to tackle the immunosuppressive barriers in therapeutic interventions, shedding light on the development of effective and personalized treatments for sarcomas. Overall, this review provides a comprehensive overview of the current status and potential of immunotherapies in sarcoma treatment, highlighting the challenges and opportunities for developing effective therapies to improve the outcomes of patients with these rare malignancies.

Construction of a Compact Array of Microplasma Jet Devices and Its Application for Random Mutagenesis of Rhodosporidium toruloides.

A small and efficient DNA mutation-inducing machine was constructed with an array of microplasma jet devices (7 × 1) that can be operated at atmospheric pressure for microbial mutagenesis. Using this machine, we report disruption of a plasmid DNA and generation of mutants of an oleaginous yeast Rhodosporidium toruloides. Specifically, a compact-sized microplasma channel (25 × 20 × 2 mm3) capable of generating an electron density of greater than 1013 cm-3 was constructed to produce reactive species (N2*, N2+, O, OH, and Hα) under helium atmospheric conditions to induce DNA mutagenesis. The length of microplasma channels in the device played a critical role in augmenting both the volume of plasma and the concentration of reactive species. First, we confirmed that microplasma treatment can linearize a plasmid by creating nicks in vitro. Second, we treated R. toruloides cells with a jet device containing 7 microchannels for 5 min; 94.8% of the treated cells were killed, and 0.44% of surviving cells showed different colony colors as compared to their parental colony. Microplasma-based DNA mutation is energy-efficient and can be a safe alternative for inducing mutations compared to conventional methods using toxic mutagens. This compact and scalable device is amenable for industrial strain improvement involving large-scale mutagenesis.

Increased expression of sodium-glucose cotransporter 2 and O-GlcNAcylation in hepatocytes drives non-alcoholic steatohepatitis.

AIMS: Steatosis reducing effects of sodium-glucose cotransporter 2 (SGLT2) inhibitors in non-alcoholic steatohepatitis (NASH) has been consistently reported in humans, but their mechanism remains uncertain. In this study, we examined the expression of SGLT2 in human livers and investigated the crosstalk between SGLT2 inhibition and hepatic glucose uptake, intracellular O-GlcNAcylation, and autophagic regulation in NASH. MATERIALS AND METHODS: Human liver samples obtained from subjects with/without NASH were analyzed. For in vitro studies, human normal hepatocytes and hepatoma cells were treated with SGLT2 inhibitor under high-glucose and high-lipid conditions. NASH in vivo was induced by a high-fat, -fructose, and -cholesterol Amylin liver NASH (AMLN) diet for 10 weeks followed by an additional 10 weeks with/without SGLT2 inhibitor (empagliflozin 10 mg/kg/day). RESULTS: Liver samples from subjects with NASH were associated with increased SGLT2 and O-GlcNAcylation expression compared with controls. Under NASH condition (in vitro condition with high glucose and lipid), intracellular O-GlcNAcylation and inflammatory markers were increased in hepatocytes and SGLT2 expression was upregulated; SGLT2 inhibitor treatment blocked these changes by directly reducing hepatocellular glucose uptake. In addition, decreased intracellular O-GlcNAcylation by SGLT2 inhibitor promoted autophagic flux through AMPK-TFEB activation. In the AMLN diet-induced NASH mice model, SGLT2 inhibitor alleviated lipid accumulation, inflammation, and fibrosis through autophagy activation related to decreased SGLT2 expression and O-GlcNAcylation in the liver. CONCLUSIONS: This study firstly demonstrates increased SGLT2 expression in NASH and secondly reveals the novel effect of SGLT2 inhibition on NASH through autophagy activation mediated by inhibition of hepatocellular glucose uptake and consequently decreased intracellular O-GlcNAcylation.

Ultrasound harmonic generation and atomic layer deposition of multilayer, deep-UV mirrors and filters with microcavity plasma arrays.

Abstract: In honor of Professor Kurt Becker's pioneering contributions to microplasma physics and applications, we report the capabilities of arrays of microcavity plasmas in two emerging and disparate applications. The first of these is the generation of ultrasound radiation in the 20-240 kHz spectral range with microplasmas in either a static or jet configuration. When a 10×10 array of microplasma jets is driven by a 20-kHz sinusoidal voltage, for example, harmonics as high as m = 12 are detected and fractional harmonics are produced by controlling the spatial symmetry of the emitter array. The preferential emission of ultrasound in an inverted cone having an angle of ±45∘ with respect to the surface normal of the jet array's exit face is attributed to interference between spatially periodic, outward-propagating waves generated by the arrays. The spatial distribution of ultrasound generated by the arrays is analogous to the radiation patterns produced by Yagi-Uda phased array antennas at RF frequencies for which radiation is emitted broadside to arrays of parallel electric dipoles. Also, the nonperturbative envelope of the ultrasound harmonic spectrum resembles that for high-order harmonic generation at optical frequencies in rare gas plasmas and attests to the strong nonlinearity provided by the pulsed microplasmas in the sub-250-kHz region. Specifically, the relative intensities of the second and third harmonics exceed that for the fundamental, and a "plateau" region is observed extending from the 5th through the 8th harmonics. A strong plasma nonlinearity appears to be responsible for both the appearance of fractional harmonics and the nonperturbative nature of the acoustic harmonic spectrum. Multilayer metal-oxide optical filters designed to have peak transmission near 222 nm in the deep-UV region of the spectrum have been fabricated by microplasma-assisted atomic layer deposition. Alternating layers of ZrO2 and Al2O3, each having a thickness in the 20-50 nm range, were grown on quartz and silicon substrates by successively exposing the substrate to the Zr or Al precursor (tetrakis(dimethylamino) zirconium or trimethylaluminum, respectively) and the products of an oxygen microplasma while maintaining the substrate temperature at 300 K. Bandpass filters comprising 9 cycles of 30-nm-thick ZrO2/50-nm-thick Al2O3 film pairs transmit 80% at 235 nm but < 35% in the 250-280 nm interval. Such multilayer reflectors appear to be of significant value in several applications, including bandpass filters suppressing long wavelength (240-270 nm) radiation emitted by KrCl (222) lamps.

Application of ionizing radiation as an elicitor to enhance the growth and metabolic activities in Chlamydomonas reinhardtii.

Chlamydomonas reinhardtii is a eukaryotic, unicellular photosynthetic organism and a potential algal platform for producing biomass and recombinant proteins for industrial use. Ionizing radiation is a potent genotoxic and mutagenic agent used for algal mutation breeding that induces various DNA damage and repair responses. In this study, however, we explored the counterintuitive bioeffects of ionizing radiation, such as X- and γ-rays, and its potential as an elicitor to facilitate batch or fed-batch cultivation of Chlamydomonas cells. A certain dose range of X- and γ-rays was shown to stimulate the growth and metabolite production of Chlamydomonas cells. X- or γ-irradiation with relatively low doses below 10 Gy substantially increased chlorophyll, protein, starch, and lipid content as well as growth and photosynthetic activity in Chlamydomonas cells without inducing apoptotic cell death. Transcriptome analysis demonstrated the radiation-induced changes in DNA damage response (DDR) and various metabolic pathways with the dose-dependent expression of some DDR genes, such as CrRPA30, CrFEN1, CrKU, CrRAD51, CrOASTL2, CrGST2, and CrRPA70A. However, the overall transcriptomic changes were not causally associated with growth stimulation and/or enhanced metabolic activities. Nevertheless, the radiation-induced growth stimulation was strongly enhanced by repetitive X-irradiation and/or subsequent cultivation with an inorganic carbon source, i.e., NaHCO3, but was significantly inhibited by treatment of ascorbic acid, a scavenger of reactive oxygen species (ROS). The optimal dose range of X-irradiation for growth stimulation differed by genotype and radiation sensitivity. Here, we suggest that ionizing radiation within a certain dose range determined by genotype-dependent radiation sensitivity could induce growth stimulation and enhance metabolic activities, including photosynthesis, chlorophyll, protein, starch, and lipid synthesis in Chlamydomonas cells via ROS signaling. The counterintuitive benefits of a genotoxic and abiotic stress factor, i.e., ionizing radiation, in a unicellular algal organism, i.e., Chlamydomonas, may be explained by epigenetic stress memory or priming effects associated with ROS-mediated metabolic remodeling.

Lysosomal control of senescence and inflammation through cholesterol partitioning.

Whereas cholesterol is vital for cell growth, proliferation, and remodeling, dysregulation of cholesterol metabolism is associated with multiple age-related pathologies. Here we show that senescent cells accumulate cholesterol in lysosomes to maintain the senescence-associated secretory phenotype (SASP). We find that induction of cellular senescence by diverse triggers enhances cellular cholesterol metabolism. Senescence is associated with the upregulation of the cholesterol exporter ABCA1, which is rerouted to the lysosome, where it moonlights as a cholesterol importer. Lysosomal cholesterol accumulation results in the formation of cholesterol-rich microdomains on the lysosomal limiting membrane enriched with the mammalian target of rapamycin complex 1 (mTORC1) scaffolding complex, thereby sustaining mTORC1 activity to support the SASP. We further show that pharmacological modulation of lysosomal cholesterol partitioning alters senescence-associated inflammation and in vivo senescence during osteoarthritis progression in male mice. Our study reveals a potential unifying theme for the role of cholesterol in the aging process through the regulation of senescence-associated inflammation.

Distal Femoral Tracker Pin Placement Prevents Delayed Pin Tract-Induced Fracture in Robotic-Assisted Total Knee Arthroplasty: Results of Minimum 1-Year Follow-Up.

The purpose of this study was to determine the outcomes of the placement of a pin tracker in the distal femur in robotic-assisted total knee arthroplasty (TKA) over a minimum follow-up period of 1 year. A consecutive series of 81 TKAs were performed in 59 patients using a robotic-assisted system (Mako) at our hospital. The mean follow-up period was 1.1 years (range, 1-1.2 years). No periprosthetic fracture at a pin tracker site was reported at a minimum of 1 year of follow-up. No patients reported minor or major complications associated with the distal femoral pins. In addition, unicortical pin-site drilling marks in all patients disappeared on one-postoperative-year follow-up radiographs. Our technique, which used unicortical pin placement in the distal femur in robotic-assisted TKA, demonstrated that it was a safe and reliable method at a minimum follow-up period of 1 year. The level of evidence is IV.

Distal Femoral Pin Tracker Placement Prevents Pin Tract-induced Fracture in Robotic-Assisted Total Knee Arthroplasty.

The purpose of this study was to determine the effectiveness of the placement of pin trackers in the medial sagittal plane of the distal femur in robotic-assisted total knee arthroplasty (TKA) over a minimum follow-up period of 3 months. From August 2020 to October 2020, a consecutive series of 81 TKAs were performed in 59 patients using the Triathlon posterior-stabilized total knee prosthesis with a robotic-assisted system (Mako) at our hospital. Patient charts were reviewed for complications associated with the pin sites, including fracture, infection, thigh pain, and the need for reoperation. No patients had any minor or major complications associated with distal femoral pins. This technique, which used pin trackers in the medial sagittal plane of the distal femur, could be a useful option for surgeons performing robotic-assisted TKA. This is a Level IV study.

Tolerogenic nanoparticles induce type II collagen-specific regulatory T cells and ameliorate osteoarthritis.

Local inflammation in the joint is considered to contribute to osteoarthritis (OA) progression. Here, we describe an immunomodulating nanoparticle for OA treatment. Intradermal injection of lipid nanoparticles (LNPs) loaded with type II collagen (Col II) and rapamycin (LNP-Col II-R) into OA mice effectively induced Col II-specific anti-inflammatory regulatory T cells, substantially increased anti-inflammatory cytokine expression, and reduced inflammatory immune cells and proinflammatory cytokine expression in the joints. Consequently, LNP-Col II-R injection inhibited chondrocyte apoptosis and cartilage matrix degradation and relieved pain, while injection of LNPs loaded with a control peptide and rapamycin did not induce these events. Adoptive transfer of CD4+CD25+ T cells isolated from LNP-Col II-R-injected mice suggested that Tregs induced by LNP-Col II-R injection were likely responsible for the therapeutic effects. Collectively, this study suggests nanoparticle-mediated immunomodulation in the joint as a simple and effective treatment for OA.

SEPHS1: Its evolution, function and roles in development and diseases.

Selenophosphate synthetase (SEPHS) was originally discovered in prokaryotes as an enzyme that catalyzes selenophosphate synthesis using inorganic selenium and ATP as substrates. However, in contrast to prokaryotes, two paralogs, SEPHS1 and SEPHS2, occur in many eukaryotes. Prokaryotic SEPHS, also known as SelD, contains either cysteine (Cys) or selenocysteine (Sec) in the catalytic domain. In eukaryotes, only SEPHS2 carries out selenophosphate synthesis and contains Sec at the active site. However, SEPHS1 contains amino acids other than Sec or Cys at the catalytic position. Phylogenetic analysis of SEPHSs reveals that the ancestral SEPHS contains both selenophosphate synthesis and another unknown activity, and that SEPHS1 lost the selenophosphate synthesis activity. The three-dimensional structure of SEPHS1 suggests that its homodimer is unable to form selenophosphate, but retains ATPase activity to produce ADP and inorganic phosphate. The most prominent function of SEPHS1 is that it is implicated in the regulation of cellular redox homeostasis. Deficiency of SEPHS1 leads to the disturbance in the expression of genes involved in redox homeostasis. Different types of reactive oxygen species (ROS) are accumulated in response to SEPHS deficiency depending on cell or tissue types. The accumulation of ROS causes pleiotropic effects such as growth retardation, apoptosis, DNA damage, and embryonic lethality. SEPHS1 deficiency in mouse embryos affects retinoic signaling and other related signaling pathways depending on the embryonal stage until the embryo dies at E11.5. Dysregulated SEPHS1 is associated with the pathogenesis of various diseases including cancer, Crohn's disease, and osteoarthritis.

Genomic Analysis of Carbapenemase-Producing Escherichia coli Isolates Analyzed by Whole Genome Sequencing.

OBJECTIVE: The increasing trend of Escherichia coli that produce extended-spectrum beta-lactamases and carbapenemases is a global public health concern. Sequence type (ST)131, a high-risk clone, is most likely responsible for the global distribution of E. coli that carry the carbapenemase gene such as bla KPC. In this study, carbapenemase-producing E. coli (CPEC) isolates were recovered from the stool of patients in a tertiary care hospital in 2019 and 2020. METHODS: We randomly selected nine isolates among CPEC isolates carrying KPC, NDM and OXA-type carbapenemase genes for whole genome sequencing (WGS). WGS was performed using the Illumina system (Illumina, Inc., San Diego, CA, USA) at Macrogen (Seoul, Korea). De novo assembly and assembly validation were performed at Macrogen. The whole genome of isolates was analyzed using the Center for Genomic Epidemiology website (http://www.genomicepidemiology.org/). RESULTS: Seven sequence types were detected: ST131, ST410, ST68, ST216, ST405, ST224 and ST409 variant. Six isolates harbored CTX-M-type genes such as bla CTX-M-15, bla CTX-M-14 and bla CTX-M-27. Plasmid replicons such as IncFIA, IncFIB, IncFII and IncX3 types were detected. CONCLUSIONS: The present study showed the diversity of the molecular characteristics of CPEC by analysis of the whole genome of bacteria. WGS is a useful tool that provides much information in the clinical microbiology.

Robot-assisted total knee arthroplasty improves mechanical alignment and accuracy of component positioning compared to the conventional technique.

PURPOSE: The purpose of this study was to compare the mechanical axis, accuracy of component positioning, and polyethylene liner thickness between robot-assisted total knee arthroplasty (TKA) and conventional TKA. METHODS: From July 2020 to December 2020, 154 TKAs were performed in 110 patients with Kellgren-Lawrence grade IV varus knees using a robot-assisted system (MAKO group). Additionally, 110 propensity score-matched patients who had undergone primary conventional TKA were chosen in a one-to-one ratio for the conventional group. Post-operative radiographs were used to evaluate mechanical axis and component coronal and sagittal positioning. The polyethylene liner thickness was investigated. The respective mean error values and outliers were obtained for the two study groups and were compared to determine the mechanical axis and the accuracy of the postoperative component positioning. RESULTS: Patients in the MAKO group achieved better accuracy than those in the conventional group in terms of postoperative mean mechanical axis (1.9˚ vs. 2.8˚, p < 0.05), femur coronal inclination (91.2˚ vs. 91.8˚, p < 0.05), tibia coronal inclination (90.8˚ vs. 91.1˚, p < 0.05), and tibia sagittal inclination (90.7˚ vs. 91.7˚, p < 0.05). However, there was no difference between the two groups in polyethylene liner thickness. CONCLUSIONS: Robot-assisted TKA showed improved mechanical axis and higher accuracy of component positioning compared to the conventional TKA technique, with no significant difference in polyethylene liner thickness between the two groups. Long-term follow-up studies are needed to compare the clinical outcomes of robot-assisted TKA. LEVEL OF EVIDENCE: IV.

Trans-Arctic vicariance in Strongylocentrotus sea urchins.

The sea urchins Strongylocentotus pallidus and S. droebachiensis first invaded the Atlantic Ocean from the Pacific following the opening of the Bering seaway in the late Miocene. While trans-Arctic dispersal during the Pleistocene is thought to have maintained species' integrity, a recent genomic analysis identified a reproductively isolated cryptic species within S. droebachiensis. Based on previous studies, the distribution of one of these lineages (S. droebachiensis W) includes the shallow water habitats of the northwest Atlantic and Pacific, while the other (S. droebachiensis E) is found throughout the shallow habitat in the northeast but is mostly restricted to deep habitats (>65 m) in the northwest Atlantic. However, since genetic variation within S. droebachiensis has been largely unstudied in the north Pacific and Arctic oceans, the biogeography of the cryptic species is not well known, and it is difficult to identify the mechanisms driving population subdivision and speciation. Here we use population genetic analyses to characterize the distribution of each species, and to test hypotheses about the role of vicariance in the evolution of systematic and genomic divergence within the genus. We collected individuals of all three Strongylocentrotus species (n = 365) from 10 previously unsampled locations in the northeast Pacific and north Atlantic (Labrador Sea and Norway), and generated mtDNA sequence data for a 418 bp fragment of cytochrome c oxidase subunit I (COI). To assess the biogeography of all three species, we combined our alignment with five previously published data sets (total n = 789) and used statistical parsimony and maximum likelihood to identify species and characterize their distribution within and among oceans. Patterns of haplotype sharing, pairwise F ST , and hierarchical analyses of molecular variance (AMOVA) identified trans-Arctic dispersal in S. pallidus and S. droebachiensis W, but other than 5 previously reported singletons we failed to detect additional mtDNA haplotypes of S. droebachiensis E in the north Pacific. Within the Atlantic, patterns of habitat segregation suggests that temperature may play a role in limiting the distribution of S. droebachiensis E, particularly throughout the warmer coastal waters along the coast of Nova Scotia. Our results are consistent with the cycles of trans-Arctic dispersal and vicariance in S. pallidus and S. droebachiensis W, but we suggest that the evolution of Atlantic populations of S. droebachiensis E has been driven by persistent trans-Arctic vicariance that may date to the initial invasion in the late Pliocene.

Mitochondrial double-stranded RNAs govern the stress response in chondrocytes to promote osteoarthritis development.

Protein kinase R (PKR) is an immune response protein that becomes activated by double-stranded RNAs (dsRNAs). PKR overactivation is associated with degenerative diseases with inflammation, including osteoarthritis (OA), but the dsRNA activator remains largely unknown. Here, we find that mitochondrial dsRNA (mt-dsRNA) expression and its cytosolic efflux are facilitated in chondrocytes under OA-eliciting conditions, leading to innate immune activation. Moreover, mt-dsRNAs are released to the extracellular space and activate Toll-like receptor 3 at the plasma membrane. Elevated levels of mt-dsRNAs in the synovial fluids and damaged cartilage of OA patients and in the cartilage of surgery-induced OA mice further support our data. Importantly, autophagy prevents PKR activation and protects chondrocytes from mitochondrial stress partly by removing cytosolic mtRNAs. Our study provides a comprehensive understanding of innate immune activation by mt-dsRNAs during stress responses that underlie the development of OA and suggests mt-dsRNAs as a potential target for chondroprotective intervention.

SPS1 deficiency-triggered PGRP-LC and Toll expression controls innate immunity in Drosophila S2 cells.

Selenophosphate synthetase 1 (SPS1) is an essential gene for the cell growth and embryogenesis in Drosophila melanogaster. We have previously reported that SPS1 deficiency stimulates the expression of genes responsible for the innate immune system, including antimicrobial peptides (AMPs), in Drosophila S2 cells. However, the underlying mechanism has not been elucidated. Here, we investigated the immune pathways that control the SPS1-deficiency-induced expression of AMPs in S2 cells. It was found that the activation of AMP expression is regulated by both immune deficiency (IMD) and the Toll pathway. Double knockdown of the upstream genes of each pathway with SPS1 showed that the peptidoglycan recognition protein-LC (PGRP-LC) and Toll genes are targeted by SPS1 for regulating these pathways. We also found that the IMD and Toll pathway regulate AMP expression by cross-talking. The levels of PGRP-LC and Toll mRNAs were upregulated upon Sps1 knockdown (6.4±0.36 and 3.2±0.45-fold, respectively, n=3). Overexpression of each protein also upregulated AMPs. Interestingly, PGRP-LC overexpression upregulated AMP more than Toll overexpression. These data strongly suggest that SPS1 controls the innate immune system of D. melanogaster through regulating PGRP-LC and Toll expression.