Input: Claim: Curcumin, found in turmeric, has anti-inflammatory properties. [SEP] Text: Staining with 4,6-diamino-2-phenyl indole (DAPI) (Sigma, St. Louis, USA) was performed to visualize nuclei. Images were obtained under a fluorescent microscope (). Real-time of brain cerebral cortex tissues was isolated using TRIzol reagent (). The expression levels of TERT mRNA were measured by real-time PCR using all-in-OneTM qPCR master mix AOPR-1200 (). The sequences of primer sets for TERT were 5’-- 3’(forward) and 5’--3’ (reverse). β-actin gene was applied as a reference. Immunohistochemistry Paraffin-embedded brain sections of 5-μm thickness were incubated with a rabbit anti-TERT antibody (Servicebio, Wuhan, China), then a biotinylated goat anti-rabbit secondary antibody (Servicebio, Wuhan, China). Immune complexes were visualized by incubation with 3,3’-diaminobenzidine tetrachloride (DAB) and hematoxylin. Measurement of parameters related to oxidative stress in cerebral homogenates The brain tissue were homogenized with 9 times the volume of PBS on ice and then centrifuged to prepare homogenates. The contents of malondialdehyde (MDA) and glutathione (GSH) as well as the activities of SOD and GSH-Px in the cerebral homogenates were measured following the respective manufacturer’s protocols (Bio-Engineering Co. , Ltd. , Nanjing, China). Protein contents in the cerebral homogenates were determined using the bicinchoninic acid assay kit (Bio-Engineering Co. , Ltd. , Nanjing, China). Measurement of IL-1β and TNF-ɑ levels in cerebral homogenates The concentrations of IL-1β and TNF-ɑ in cerebral homogenates were determined using using ELISA kits according to the manufacturer’s instructions (IL-1β: PeproTech Inc. , NJ, USA TNF-ɑ: R&). Statistical analysis All experiments were conducted with two technical replicates.

Output: pass


Input: Claim: The majority of research in graphene focuses on its application in supercapacitors and energy storage due to its high electrical conductivity. [SEP] Text: [44] F. Zhang, T. Zhang, X. Yang, L. Zhang, K. Leng, Y. Huang, Y. Chen, A high-performance supercapacitor-battery hybrid energy storage device based on graphene-enhanced electrode materials with ultrahigh energy density, Energy Environ. Sci. 6 (2013) 1623–1632. [45] Y. K. A. Lau, A. V Kvit, A. L. Schmitt, S. . . Growth, S. Publishing, N. Cabrera, C. . . Keis, S. E. Lindquist, A. . Dudley, D. Bliss, M. Callahan, M. . Yoreo, C. Cottrell, S. Award, D. Young, Monolithic Carbide-Films for Micro-Supercapacitors, Science 1060 (2010) 480–484. [46] W. Sun, X. Chen, Fabrication and tests of a novel three dimensional micro supercapacitor, Microelectron Eng. 86 (2009) 1307-1310. [47] W. Sun, R. Zheng, X. Chen, Symmetric redox supercapacitor based on micro-fabrication with three-dimensional polypyrrole electrodes, J. 195 (2010) 7120–7125. [48] C. Shen, X. Wang, W. Zhang, F. Kang, A high-performance three-dimensional micro supercapacitor based on self-supporting composite materials, J. 196 (2011) 10465–10471. [49] M. Beidaghi, Y. Gogotsi, Capacitive energy storage in micro-scale devices: Recent advances in design and fabrication of micro-supercapacitors, Energy Environ. Sci. 7 (2014) 867–884. [50] M. Beidaghi, C. Wang, Micro-supercapacitors based on interdigital electrodes of reduced graphene oxide and carbon nanotube composites with ultrahigh power handling performance, Adv. Funct. Mater. 22 (2012) 4501–4510. [51] J. Sung, S. Kim, K.

Output: corroborate


Input: Claim: Ocean acidification only affects animals with hard shells, and fish are immune. [SEP] Text: Figures AbstractOcean acidification (OA) threatens marine crustaceans, particularly those at high latitudes where conditions are rapidly changing. Red king crab (RKC) support important fisheries in Alaskan waters. RKC early life stages are sensitive to OA when exposure occurs as embryos and juveniles. However, in a supporting study, RKC were surprisingly tolerant of varying OA conditions (pH 7. 5, 7. 8, & 8. 0) when reared long-term from larval hatching to the first crab stage (C1). Here, we examined gene expression in the C1 juveniles to characterize transcriptional activity of these OA-tolerant crabs. Expression of nearly half of all genes (44%) correlated with OA treatment, suggesting a strong molecular response to OA, contrary to the phenotypic results. Downregulated functions were numerous in response to OA, and included reduced energy production, biosynthesis, immune function, and notably lipid and carbohydrate metabolic processes, which suggest a shift in metabolic strategy to protein catabolism, possibly to reduce CO2 production and facilitate acid/base regulation. Only a handful of functions were enriched in OA-induced genes, which were related to transcription regulation, control of growth and cell death, and signaling activity. Transposon activity was high in OA-reared crab, many of which were upregulated at consistent levels, suggesting that transposon mobilization may be a component of the RKC OA-response system. Genetic composition did not differ among OA treatments indicating that transcriptional differences in OA-reared crab were more likely attributed to phenotypic plasticity than selective mortality. Our results suggest that OA-reared RKC have a narrowed, possibly optimized, set of gene functions that enables OA-tolerance through the early juvenile stage. OA-exposure from hatch may benefit RKC and related species by “hardening” them against OA through physiological reprogramming. Future studies should test OA-hardened crabs in additional challenges, as metabolic and immune limitations may ultimately make them more vulnerable to infection or secondary stressors. Citation: (2024) Narrowed gene functions and enhanced transposon activity are associated with high tolerance to ocean acidification in a juvenile subarctic crustacean. PLOS Clim 3(3): e0000319.

Output: contrast


Input: Claim: Curcumin, found in turmeric, has anti-inflammatory properties. [SEP] Text: AbbreviationsACR: Acrylamide; GSH: Glutathione; GSH-Px: Glutathione peroxidase; HE: Hematoxylin and eosin; IL-1β: Interleukin-1β; MDA: Malondialdehyde; ROS: Reactive oxygen species; SOD: Superoxide dismutase; TERT: Telomerase reverse transcriptase; TNF-α: Tumor necrosis factor-α; TUNEL: Terminal deoxynucleotidyl transferase mediated dUTP nick end labellingDeclarationsEthics approval and consent to participate Animal experiments were approved by the of Medicine College, of Science and Technology (resolution number 2019078), and accomplished in line with the guidelines of the Council of China. Consent for publication Not applicable. Availability of data and materials The datasets supporting the conclusions of this 57. . Acrylamide in human diet, its metabolism, toxicity, inactivation and the associated legal regulations in food industry. Nutr. 2019 Mar 25:1-16. . Calpeptin is neuroprotective against acrylamide-induced neuropathy in rats. Toxicology. 2018;400-401:1-8. . Neurotoxicity of acrylamide in exposed workers. . 2013; 10(9):3843-54. . Down-regulation of telomerase reverse transcriptase-related anti-apoptotic function in a rat model of acrylamide induced neurobehavioral deficits. Biotech Histochem. 2018;93(7):512-8. . Acrylamide exerts its cytotoxicity in NIH/3T3 fibroblast cells by apoptosis. Health. 2018;34(7):481-9. . Acrylamide-induced apoptosis in rat primary astrocytes and human astrocytoma cell lines. Vitro. 2014;28(4):562-70. . Taurine attenuates acrylamide- induced apoptosis via a PI3K/AKT-dependent manner. Toxicol. 2018;960327118765335. . Roles for Telomerase in the System. Neurosci. 2018;11:160. .

Output: pass


Input: Claim: The majority of research in graphene focuses on its application in supercapacitors and energy storage due to its high electrical conductivity. [SEP] Text: Zhou, X. S. Zhao, Graphene-based materials as supercapacitor electrodes, J. Mater. Chem. 20 (2010) 5983–5992. [71] E. Frackowiak, Carbon materials for supercapacitor application, 9 (2007) 1774–1785. [72] L. M. Da Silva, L. A. De Faria, J. F. C. Boodts, Determination of the morphology factor of oxide layers, Electrochim. Acta 47 (2001) 395–403. [73] G. Yu, L. Hu, M. Vosgueritchian, H. Wang, X. Xie, J. R. McDonough, X. Cui, Y. Cui, Z. Bao, Solution-processed graphene/MnO2 nanostructured textiles for high-performance electrochemical capacitors, Nano Lett. 11 (2011) 2905–2911. [74] A. Borenstein, O. Hanna, R. Attias, S. Luski, T. Brousse, D. Aurbach, Carbon-based composite materials for supercapacitor electrodes: A review, J. Mater. Chem. A 5 (2017) 12653–12672. [75] L. Lv, Y. Fan, Q. Chen, Y. Zhao, Y. Hu, Z. Zhang, N. Chen, L. Qu, Three-dimensional multichannel aerogel of carbon quantum dots for high-performance supercapacitors, Nanotechnology 25 (2014) 235401. [76] Y. Wang, A. Hu, Carbon quantum dots: Synthesis, properties and applications, J. Mater. Chem. C 2 (2014) 6921–6939. [77] Y. -Q. Dang, S. -Z. Ren, G. Liu, J. Cai, Y. Zhang, J. Qiu, Electrochemical and capacitive properties of carbon dots/reduced graphene oxide supercapacitors, Nanomaterials 6 (2016) 212. [78] X. Chen, R. Paul, L. Dai, Carbon-based supercapacitors for efficient energy storage, Natl. Sci. Rev.

Output: corroborate


Input: Claim: Ocean acidification only affects animals with hard shells, and fish are immune. [SEP] Text: for : October 18, 2023; Accepted: February 22, 2024; Published: March 28, 2024This is an open access 7324. Code and analysis files used in this study are available in the accompanying repository ( This work was supported by the 1530-1213 (to WCL and IBS) and 21404 (to WCL and IBS). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Competing interests: The authors have declared that no competing interests exist. IntroductionGlobal oceans are rapidly changing due to increased concentrations of atmospheric greenhouse gasses. In addition to warming, deoxygenation, and sea level rise, oceans are acidifying [1]. This is occurring due to the increased partial pressure of carbon dioxide (CO2) in the ocean, which has absorbed ~30% of added atmospheric CO2 emitted since the industrial revolution [2]. As a result, the concentration of aqueous CO2, hydrogen ions, and bicarbonate ions are increasing, and ocean pH, carbonate ion concentrations, and calcium carbonate saturation states are decreasing. This shift is referred to as ocean acidification (OA) [3], and, in addition to the effects of lower pH, is particularly concerning for calcifying species including crustaceans, molluscs, and corals due to their reliance on calcium carbonate (aragonite, calcite) to form shells and exoskeletons [4, 5]. While OA is a global phenomenon, changes in high latitude coastal zones are likely to have outsized biological effects. Carbon dioxide is more soluble at high latitudes due to colder water, resulting in lower pH levels and carbonate saturation states [6, 7]. Increased freshwater inputs from sea ice melt and river runoff, and enhanced upwelling and respiration are likely to augment acidification in high latitude coastal zones [6]. Conditions in those regions are therefore more likely to become undersaturated with carbonate ions and reach severely low pH levels [7, 8]. Sea shelf, a coastal area off Alaska which supports highly productive commercial, subsistence, and sport fisheries [9], already experiences seasonal levels of aragonite saturation (Ω) below one, which is the biological threshold for calcium carbonate dissolution [10–13].

Output: contrast


Input: Claim: Curcumin, found in turmeric, has anti-inflammatory properties. [SEP] Text: -19 PreprintsProtocolsVideosJournalsTools & a 10. 21203/rs. 3. rs-128652/v1This work is licensed under a CC BY 4. 0 LicenseStatus:Under ReviewVersion 1posted 9You are reading this latest preprint versionAbstractPAC (3,5-Bis (4-hydroxy-3-methoxybenzylidene)-N-methyl-4-piperidone), a novel bioactive curcumin analog, has been reported to have anticancer properties against various tumors. However, the anti-cancer effects of PAC on oral cavity squamous cell carcinoma (OC-SCC) were not studied yet. Our aim is to investigate the anti-oral cancer properties of PAC in vitro, and determine the molecular mechanisms underlying these effects. Viability assays including MTT and LDH were conducted to measure cell proliferation. Flow cytometry-based cytotoxicity assay was performed to detect autophagic cell death and oxidative stress markers. Western blotting was used for measuring protein expression/activation in apoptotic, autophagic and pro-carcinogenic cellular signaling pathways. We demonstrated that PAC preferentially and, in a dose, -dependent way kills oral cancer cells, but was not toxic to normal human gingival cells. PAC destabilizes cell-cycle distributions, inhibits the expression of oncogenes (cyclin D1) and that of cyclin-dependent kinase inhibitor (p21WAF1) is upregulated, increases the expression of p53 gene, and inhibits epithelial-mesenchymal transition markers in oral cancer cells. The PAC effect involve various signaling pathways including NF-κcaspase-3/9 and PARP1. Finally, PAC demonstrated ability to induce autophagy, decrease production of reactive oxygen species (ROS), increase intracellular glutathione (GSH) activity, and reduce mitochondrial membrane potential in oral cancer cells. In conclusion, PAC inhibits the proliferation and increases the apoptosis and autophagy and oxidative stress of oral cancer cells. These effects involve ERK1/2, p38/JNK, NF-κB and Wnt cellular signaling pathways. Overall, our study suggests the potential use of PAC to treat oral cancer.

Output: pass


Input: Claim: The majority of research in graphene focuses on its application in supercapacitors and energy storage due to its high electrical conductivity. [SEP] Text: Am. Chem. Soc. 131 (2009) 11308–9. [87] Q. Wang, X. Liu, L. Zhang, Y. Lv, Microwave-assisted synthesis of carbon nanodots through an eggshell membrane and their fluorescent application, Analyst 137 (2012) 5392–5397. [88] H. Jiang, F. strategy for synthesis and functionalization of carbon nanop1995. [89] X. Xu, R. Ray, Y. Gu, H. J. Ploehn, L. Gearheart, K. Raker, W. A. Scrivens, Electrophoretic analysis and purification of fluorescent single-walled carbon nanotube fragments, J. Am. Chem. Soc. 126 (2004) 12736–12737. [90] J. Xu, K. Hou, Z. Ju, C. Ma, W. Wang, C. Wang, J. Cao, Z. Chen, Synthesis and electrochemical properties of carbon dots/manganese dioxide (CQDs/MnO2) nanoflowers for supercapacitor applications, J. Electrochem. Soc. 164 (2017) A430–A437. [91] Y. Zhu, X. Ji, C. Pan, Q. Sun, W. Song, L. Fang, Q. Chen, C. E. Banks, A carbon quantum dot decorated RuO2 network: Outstanding supercapacitances under ultrafast charge and discharge, Energy Environ. Sci. 6 (2013) 3665–3675. [92] H. Feng, P. Xie, S. Xue, L. Li, X. Hou, Z. Liu, D. Wu, L. Wang, P. K. Chu, Synthesis of three-dimensional porous reduced graphene oxide hydrogel/carbon dots for high-performance supercapacitor, J. Electroanal. Chem. 808 (2018) 321–328. [93] S. Mondal, U. Rana, S. Malik, Graphene quantum dot-doped polyaniline nanofiber as high performance supercapacitor electrode materials, Chem. Comm. 51 (2015) 12365–12368.

Output: corroborate


Input: Claim: Ocean acidification only affects animals with hard shells, and fish are immune. [SEP] Text: Model projections for the through the year 2100 predict longer seasonal periods of Ω<1 (up to five and two months in the winter for aragonite and calcite, respectively) [14, 15, pers. comm. ], and that pH will decrease 0. 07–0. 35 pH units in surface waters and 0. 05–0. 30 in bottom waters [14, 15]. These carbonate changes have significant socio-economic implications for Alaskan crustaceans, given their importance to fisheries as both prey species (krill, copepods) [16, 17] and fishery stocks (e. g. king, tanner, and snow crabs) [18, 19]. OA has broad but variable physiological effects on crustaceans [20]. Many are capable of maintaining hemolymph pH homeostasis by acid-base regulation [21, 22], and have exoskeletons composed of both calcium carbonate and chitin, which may protect against direct dissolution [23, 24]. Still, tissue and shell growth [25], molt cycle [26, 27], exoskeleton properties [28, 29], metabolic activity [30, 31], reproduction [22, 32], behavior [33, 34], and immune function [22, 35] can all be altered by OA, particularly at higher pCO2 concentrations [36]. Effects are most acutely observed in early life stages (larvae, juveniles), and vary greatly by species, severity of OA, life stage, and duration of exposure [20, 28, 37, 38]. The diversity of responses in crustaceans, and all marine invertebrates for that matter, has highlighted the need for a more mechanistic understanding of how organisms function in OA conditions. Targeted gene expression analyses in green shore crab (Carcinus maenas) gill tissue reported that genes involved in acid-base regulation are affected by high pCO2 [39, 40]. Transcriptome-wide expression analysis identified changes in genes involved in energy metabolism and apoptosis activity in shrimp (Exopalaemon carinicauda) [41], and immune functions, energy metabolism, and ion transport in the Chinese mitten crab (Eriocheir sinensis) [42, 43].

Output: contrast


Input: Claim: Curcumin, found in turmeric, has anti-inflammatory properties. [SEP] Text: Cancer BiologyOncologyHead & Neck SurgeryPACoral cancerapoptosisautophagyoxidative stressSignaling pathwaysFiguresFigure 1Figure 2Figure 3Figure 4Figure 5Figure 6Figure 7Figure 8Figure 9Figure 10cancer is commonly known as one of the head and neck cancers. It is the sixth most common malignancy worldwide 1. According to the Society in 2019, approximately 5,300 people will be diagnosed with oral cavity cancer, which 3,700 cases concerned men and 1,600 are women. This cancer will cause death of 1,450 Canadians, which 1,050 are men and 450 women 2. Oral cancer is critical, because the patient may not notice it in its first stages, as it can frequently thrive without generating pain or symptoms, and generally, it produces primary tumors. This means that patients, who survive a first encounter with the disease, have up to 20 times higher risk of developing a second cancer3. Largely, those who have been suffering from buccal mucosa cancer for 5 years have about a 50 percent life expectancy or mortality rate4). Treating oral cancer conventionally is highly dependent on surgery assisted with or without chemotherapy and radiotherapy. For destroying malignant cells, chemicals are used more extensively as a systemic treatment 5. The panel of anti- cancer drugs includes mainly 5-fluorouracil, hydroxyurea, platinum derivatives, anthracyclines, plant alkaloids6. The most common chemotherapy medicines consists of cisplatin 7. Despite its survival benefits as an anticancer drug, it has side effects, especially the chemoresistance that is limiting patients in clinical effectiveness 7. This knowledge has inspired the use of herbal medicinal products as complementary and/or alternative therapies to a direct remedy, thus the recognition of plant products as effective and inexpensive sources of novel synthetic chemotherapeutic compounds is increasing. For decades, in some countries such as China, India, and Egypt, traditional medicine has been making use of flavored herbs in its home-based therapies for many diseases, including cancer. Nowadays, it is becoming more widely acceptable to consider herbal remedies as helpful supplements to medical prescriptions 8; 9 10. Curcumin, a yellow shade from Curcuma longa Linn. , is a significant part of turmeric and is normally used as a flavor and nourishment shading material11).

Output: pass


Input: Claim: The majority of research in graphene focuses on its application in supercapacitors and energy storage due to its high electrical conductivity. [SEP] Text: [94] Z. Zhao, Y. Xie, Enhanced electrochemical performance of carbon quantum dots-polyaniline hybrid, J. 337 (2017) 54–64. [95] X. Zhang, J. Wang, J. Liu, J. Wu, H. Chen, H. Bi, Design and preparation of a ternary composite of graphene oxide/carbon dots/polypyrrole for supercapacitor application: Importance and unique role of carbon dots, Carbon 115 (2017) 134–146. [96] L. Ferragut, R. Montenegro, G. Winter, A. Núñez, Accurate extraction of interconnect capacitances by adaptive mixed F. E. M. , Microprocessing and Microprogramming 32 (1991) 61–68. [97] C. Niu, E. K. Sichel, R. Hoch, D. Moy, H. Tennent, High power electrochemical capacitors based on carbon nanotube electrodes, Appl. Phys. Lett. 70 (1997) 1480–1482. [98] R. Z. Ma J. . Liang, B. Q. Wei, B. Zhang, C. L. Xu, Study of electrochemical capacitors utilizing carbon nanotube electrodes, J. 84 (1999) 126–129. [99] B. Hsia, J. Marschewski, S. Wang, J. Bin In, C. Carraro, D. Poulikakos, C. P. Grigoropoulos, R. Maboudian, Highly flexible, all solid-state micro-supercapacitors from vertically aligned carbon nanotubes, Nanotechnology 25 (2014) 055401. [100] Y. Q. Jiang, Q. Zhou, L. Lin, Planar mems supercapacitor using carbon nanotube forests, Proceedings of the on (MEMS), (2009). [101] W. Yu, H. Zhou, B. Q. Li, S. Ding, 3D Printing of carbon nanotubes-based microsupercapacitors, ACS Appl. Mater. Interfaces 9 (2017) 4597–4604. [102] A. K. Geim and K. S.

Output: corroborate


Input: Claim: Ocean acidification only affects animals with hard shells, and fish are immune. [SEP] Text: Metabolomic analyses in juvenile Dungeness (Cancer magister) and green shore (Carcinus maenas) crabs both found amino acid metabolism to respond to acidified conditions, perhaps due to increased buffering needs [44, 45]. As with biometric responses, molecular effects of OA can vary by life stage. Metabolite analyses of American lobster larvae (Homarus americanus) indicate that large metabolic shifts (fatty acids, amino acids, and citrate cycle) perhaps underlie their buffering capacity and tolerance of OA, whereas metabolic reprogramming is not observed in the more sensitive juvenile stage [46]. Together, molecular assays to date reveal that OA induces changes in energy metabolism, acid-base regulation, immune function, and cellular stress-response processes, with the strongest responses perhaps reflective of physiological reprogramming. Changes are, however, somewhat unpredictable due to species-, stage-, and exposure-specific effects. For that reason, the most informative studies pair molecular assays with survival and growth data preferably from the same individuals. Paralithodes camtschaticus, red king crab, is one of several crab species that are a highly valued fishery in Alaskan waters (17M pounds, or $90M USD, in annual commercial landings on average from 2000–2022, for all king crab species [47]) but that may be threatened by ocean acidification [18]. Early life stages, which are thought to be particularly vulnerable to OA, are present as brooded embryos year-round and from hatch in late spring through settlement in fall [48, 49] and may encounter low carbonate saturation states that already occur in bottom waters and during seasonal carbonate cycles [13]. Laboratory studies on embryos, early zoea, and juveniles indicate that OA negatively affects early red king crab life stages such that development is altered, and growth and survival typically decreases [50–52]. OA also interacts with warming by increasing intermolt duration and decreasing survival in juveniles [53].

Output: contrast


Input: Claim: Curcumin, found in turmeric, has anti-inflammatory properties. [SEP] Text: Furthermore, it was established that PAC is also repressing NF-kB and its downstream effectors cyclin D1 and Bcl-2 (B-cell lymphoma 2) as well as JAK2/STAT3, AKT/mTOR (mammalian target of rapamycin) and MEK/ERK (mitogen-activated protein kinase/extracellular signal-regulated kinase) signaling pathways 23 21; 24. It was demonstrated that PAC is a potent inhibitor of these pathways and has better solubility and biodistribution than curcumin 25. Until today, no study was available to investigate its properties to possibly treat oral cancer. This research is aiming at studying the inhibitory effects of the curcumin analog PAC on oral cell proliferation and apoptosis, as well as investingating the signaling pathways involved in the interactions of PAC with oral cancer cells. 5-Bis (4-hydroxy-3-methoxybenzylidene)-N-methyl-4-piperidone (PAC) was obtained from Dr. Ibrahim Al-Jammaz’s laboratory (Riyadh, Saudi Arabia). Ca9-22 cell line was purchased from (Tsukuba-shi, Japan). However, primary human gingival epithelial cells (GEC) and primary gingival fibroblasts (GF), were isolated from healthy human gingival tissues as we previously reported. committee of approved the use of primary human gingival cells. The gingival cells were extracted after the donor consent. RPMI-1640, F12, DMEH cell culture media, fetal calf serum (FCS), fetal bovine serum (FBS) and have been bought from Scientific (). Penicillin-Streptomycin, MTT and LDH colorimetric assays, EDTA solution, Caspases detection kit (FITC-VAD-FMK) and β-actin antibody (A5441) were all from Sigma-Aldrich (Oakville, Ontario, Canada). with propidium iodide was obtained from BD Biosciences (Mississauga, ON, Canada). (Red), Species (ROS) (GSH) Assay were purchased from Immuno(Burlington, ON, Canada).

Output: pass


Input: Claim: The majority of research in graphene focuses on its application in supercapacitors and energy storage due to its high electrical conductivity. [SEP] Text: Novoselov, The rise of graphene, Nat. Mater. 6 (2007) 183–191. [103] J. Xia, F. Chen, J. Li, N. Tao, Measurement of the quantum capacitance of graphene, Nat. Nanotechnol. 4 (2009) 505–509. [104] X. Cao, Z. Yin, H. Zhang, Three-dimensional graphene materials: Preparation, structures and application in supercapacitors, Energy Environ. Sci. 7 (2014) 1850–1865. [105] S. . Govindaraj, C. N. R. Rao, Graphene-based electrochemical supercapacitors, J. Chem. Sci. 120 (2008) 9–13. [106] Y. Wu, T. Zhang, F. Zhang, Y. Wang, Y. Ma, Y. Huang, Y. Liu, Y. Chen, In situ synthesis of graphene/single-walled carbon nanotube hybrid material by arc-discharge and its application in supercapacitors, 1 (2012) 820–827. [107] J. M. Chem, C. M. Ma, C. Hu, Design and tailoring of a hierarchical graphene-carbon nanotube architecture for supercapacitors J. Mater. Chem. 21 (2011) 2374-2380. [108] J. Li, M. Östling, Prevention of graphene restacking for performance boost of supercapacitors—A review, Crystals 3 (2013) 163–190. [109] Z. Gao, W. Yang, J. Wang, B. Wang, Z. Li, Q. Liu, M. Zhang, L. Liu, A new partially reduced graphene oxide nanosheet/polyaniline nanowafer hybrid as supercapacitor electrode material, 27 (2013) 568–575. [110] Z. Li, H. Zhang, Q. Liu, L. Sun, L. Stanciu, J. Xie, Fabrication of high-surface-area graphene/polyaniline nanocomposites and their application in supercapacitors, ACS Appl. Mater. Interfaces 5 (2013) 2685–2691.

Output: corroborate


Input: Claim: Ocean acidification only affects animals with hard shells, and fish are immune. [SEP] Text: One study to date has explored molecular changes in OA-exposed red king crab, and found no pronounced effects on the larval transcriptome (individuals were pooled) [54] which corresponded with decreased survival rates [52]; whereas in juveniles, expression of genes involved in cuticular processes were strongly affected by OA exposure [54] despite no changes in morphology (survival and growth were negatively affected) [51]. Interestingly, Long et al. [55] recently found that when red king crabs are exposed to OA during multiple stages of development—from hatch through the early juvenile stage—they are surprisingly tolerant of moderate (pH 7. 8) and severe (pH 7. 5) acidification, with no change in survival, growth, or development [42]. Therefore, when OA-exposure is prolonged and begins at hatch, red king crab may effectively acclimate without negative physiological impacts. The mechanisms that enable OA tolerance are of profound interest to those that manage and rely on the red king crab fishery. In this study, we used functional genomics to examine the molecular response of OA-exposed red king crab. We leveraged juveniles from Long et al. [55] that were exposed to (and tolerated) three carbonate chemistry treatments from hatching to the first crab stage (C1), thus capturing transcriptional differences among crab that are reared in historically ambient conditions along the shelf (pH 8. 0), and those acclimated to a moderately (pH 7. 8) and severely (pH 7. 5) acidified environments that are projected to occur in surface and bottom waters by the end of this century [15]. Using RNA-Seq, a high-throughput sequencing approach that measures gene-activity, our study provides a snap-shot of system-wide changes in energy allocation due to acidification exposure by identifying genes, their functions, and biological processes that differ in OA-reared crab [56]. Libraries were constructed from at least 13 individuals per treatment, rather than pools of individuals which can obscure genotype-dependent variation. Importantly, since the crab used in this experiment were quite tolerant of OA conditions [42], the molecular mechanisms and pathways described here may be potentially critical to survival in an acidified environment. statement Red king crabs are non-cephalopod invertebrates and research involving them is exempt from ethics approval.

Output: contrast


Input: Claim: Curcumin, found in turmeric, has anti-inflammatory properties. [SEP] Text: Blue G-250 Dye has been bought from Bio-Rad (Mississauga, ON, Canada), while PVDF membranes were from Cytiva (Vancouver, BC, Canada). The following antibodies, among which p53 (sc-263), cyclin D1 (sc-8396), p21 (sc-6246), Bcl-2 (sc-509), Bax (sc-7480), PARP-1 (sc-8007), caspase-3 (sc-56046), caspase-9 (sc-17784), NF-kB (sc-8008) and β-catenin (sc-59737) were all obtained from Biotechnology (). Anti-vimentin antibody (Ab8978) from Abcam () as well as others to cleaved caspase-3 (9664S), cleaved caspase-9 (20750S), cleaved PARP-1 (5625S), cytochrome C (11940S), E-cadherin (8834), pERK1/2 (4370), ERK1/2 (4695), pp38 MAPK (4631), p38 MAPK (9212), p-SAPK/JNK (4668P), SAPK/JNK (9252S), LC3B (2775) and SQSTM1/p62 (39749) were all from Technology (). The goat anti-mouse (554002) and anti-rabbit (554021) secondary and have been bought from BD Pharmingen (Mississauga, ON, Canada). and PAC Stimulation Ca9-22 cells and GEC were sub-cultured respectively in RPMI-1640, F12 or DMEM media supplemented with 5% (v/v) FBS for Ca9-22 and 10% for GEC, 100 units/mL penicillin, and 100 µg/mL streptomycin in T75 flasks 26; 27. All cells were maintained in a humidified atmosphere of 5% CO2 at 37 °C. All experiments conducted with GEC were at passages 3 to 4, and GF at passages 4 to 5. PAC was used at (0, 1, 2.

Output: pass


Input: Claim: The majority of research in graphene focuses on its application in supercapacitors and energy storage due to its high electrical conductivity. [SEP] Text: [111] J. M. Chem, High performance supercapacitors using metal oxide anchored graphene nanosheet electrodes, J. Mater. Chem. 21 (2011) 16197–16204. [112] J. Wang, Z. Gao, Z. Li, B. Wang, Y. Yan, Q. Liu, T. Mann, Journal of synthesis of graphene nanosheets ZnO composites and electrochemical properties, J. Chem. 184 (2011) 1421–1427. [113] Z. Gao, J. Wang, Z. Li, W. Yang, B. Wang, M. Hou, Y. He, Q. Liu, T. Mann, P. Yang, M. Zhang, L. Liu, Graphene Nanosheet/Ni2+/Al3+ Layered Double-as a for a Supercapacitor, Chem. Mater. 23 (2011) 3509–3516. [114] W. Gao, N. Singh, L. Song, Z. Liu, A. L. M. Reddy, L. Ci, R. Vajtai, Q. Zhang, B. Wei, P. M. Ajayan, Direct laser writing of micro-supercapacitors on hydrated graphite oxide films, Nat. Nanotechnol. 6 (2011) 496–500. [115]. Majumder, Miniaturized Supercapacitors: with Metrics, Adv. Energy Mater. 5 (2015) (1-10). [116] M. F. El-Kady, R. B. Kaner, Scalable fabrication of high-power graphene micro-supercapacitors for flexible and on-chip energy storage, Nat. Commun. 4 (2013) (1-7). [117] D. Shen, G. Zou, L. Liu, W. Zhao, A. High-Graphene Micro-Supercapacitors by a Writing and. Mater. Interfaces 10 (2018) 5404–5412. [118] J. Lin, Z. Peng, Y. Liu, F. Ruiz-Zepeda, R. Ye, E. -induced porous graphene films from commercial polymers, Nat.

Output: corroborate


Input: Claim: Ocean acidification only affects animals with hard shells, and fish are immune. [SEP] Text: Briefly, survival, developmental time, calcification, and mass did not differ among treatments at any stage, nor did survival or developmental time cumulatively from hatch to the C1 juvenile stage differ. Genetic relatedness analysis Principal component analysis (PCA) constructed from SNPs (n = 331) indicated genetic homogeneity among treatments (Fig 1, S2 Fig). Individuals loosely aggregated into two or three clusters, primarily along PC1 and PC2 which explained 8. 7% and 7. 3% of variation, respectively, but no treatments were overrepresented in any of the clusters. Pairwise FST values among samples from the three treatments did not differ from zero for any contrast (Table 3). Parentage analysis using Colony estimated the same number of mothers (n = 9) and fathers (n = 10) represented by surviving offspring in each treatment. No correlation was found among SNP-derived PCA scores and gene expression-derived PCA scores along the first two principal components (S3 Fig). Download: PPTPowerPoint slidePNGlarger imageTIFForiginal imageFig 1. PCA biplot of the first two principal components, constructed from RNA-Seq derived SNPs (n = 331). Points represent individual crabs that are color-coded by OA treatment, which do not indicate clustering by treatment, and sizes represent the percent of SNPs that are missing in each individual. The PCA and other genetic analyses (parentage, pairwise FST) does not indicate that genetic composition of individuals surviving the three-month exposure differed among treatments. Download: PPTPowerPoint slidePNGlarger imageTIFForiginal imageTable 3. Pairwise FST values. RNA-Seq pre-processing for gene expression analysis A total of ~2. 65B paired-end reads (henceforth “fragments”) remained after discarding ~4. 55M (0. 17%) during initial quality-filtering, with a per-sample mean of 61. 7M +/- 10. 7M fragments. Across all samples ~2. 14B reads were aligned to the P. camtschaticus draft genome [65, 66], for a total alignment rate of 80. 47%. The average per-sample alignment rate was 80. 43%±2. 15%, which included 43. 19%±1. 50% and 24. 52%±0.

Output: contrast


Input: Claim: Curcumin, found in turmeric, has anti-inflammatory properties. [SEP] Text: 5, 5 and 10 µM in DMSO) with all experiments, except for Western blotting where we used PAC at 5 µM, only. Assay: Cells were seeded at 3 × 105 cells/well in 6-well cell culture plates, and culture for 24 h. Culture were then stimulated exposed to vehicle control (DMSO) or various concentrations of PAC (0, 1, 2. 5, 5 and 10 µM) for 24 h. At the end of the stimulation period, the culture medium of each condition was supplemented with MTT solution at 0. 5 mg/mL then incubated for 3 h at 37ºC in the dark. The cells were washed, then 1 ml of 0. 04 N HCl in isopropanol was added to the culture wells, followed by an extended 15-min incubation. Finally, 200 µl of the reaction mixture (in triplicate) was transferred to the wells of a 96-well flat-bottom plate and the absorbance was measured at 550 nm using iMark reader (Bio-Rad). The cell viability (percentage) was determined by using the following formula: % of cell viability = [(OD550 nm (treated cell)/(OD (control cell)] × 100. This experiment was repeated 7 times. To support the cell viability assay we performed lactate dehydrogenase (LDH) activity measurement using culture supernatants. Supernatants were collected from each condition after exposure or not of the cells to PAC for 24 h. The LDH activity was measured by means of an LDH kit from (Sigma-Aldrich), as outlined in our previous work 28. The experiment was repeated 4 independent times. Assay Ca9-22 cells were seeded into 6-well plates at 103 cells per well for 24 h, before being treated with various concentrations of PAC. The culture medium was changed every 2 or 3 days. After 2 weeks, the cell clones that survived were fixed by 100% ethanol for 10 min. After washing twice by PBS, cells were then stained with 0. 5% crystal violet for 30 min and they were photographed with a digital camera. Annexin V-with propidium iodide Ca9-22 cells were seeded into T25 flasks the exposed to 1 µM, 2. 5 µM, 5 µM or 10 µM of PAC, for 24 h and 48 h, at 37 °C.

Output: pass


Input: Claim: The majority of research in graphene focuses on its application in supercapacitors and energy storage due to its high electrical conductivity. [SEP] Text: Commun. 5 (2015) 5–12. [119] J. Luo, F. R. Fan, T. Jiang, Z. Wang, W. Tang, C. Zhang, M. Liu, G. Cao, Z. L. Wang, Integration of micro-supercapacitors with triboelectric nanogenerators for a flexible self-charging power unit, Nano Res. 8 (2015) 3934–3943. [120] Z. Peng, R. Ye, J. A. Mann, D. Zakhidov, Y. Li, P. R. Smalley, J. Lin, J. M. Tour, Flexible boron-doped laser-induced graphene microsupercapacitors, ACS Nano 9 (2015) 5868–5875. [121] S. Wang, Z. Wu, S. Zheng, F. Zhou, C. Sun, H. Cheng, X. Bao, Scalable fabrication of photochemically reduced graphene-based monolithic micro- supercapacitors with superior energy and power densities, ACS Nano 11 (2017) 4283−4291. [122] Z. Liu, Z. Wu, S. Yang, R. Dong, X. Feng, K. Müllen, Ultraflexible in-plane micro-supercapacitors by direct printing of solution-processable electrochemically exfoliated graphene, Adv. Mater. 28 (2016) 2217–2222. [123] J. Li, S. S. Delekta, P. Zhang, S. Yang, M. R. Lohe, X. Zhuang, X. Feng, Scalable fabrication and integration of graphene microsupercapacitors through full inkjet printing, ACS Nano 11 (2017) 8249−8256. [124] G. Xiong, C. Meng, R. G. Reifenberger, Graphitic petal micro-supercapacitor electrodes for ultra-high power graphitic petal micro-supercapacitor electrodes for ultra-high power density, Energy Technol. 2 (2014) 897-905. Related products Composite Materials, 15. 95 Materials, 15. 95 Materials, 15.

Output: corroborate


Input: Claim: Ocean acidification only affects animals with hard shells, and fish are immune. [SEP] Text: 50), which is evident from the biplot of the first two principal components (PC1 & PC2) (Fig 2). Ambient-reared crabs were separated from those reared in OA treatments along PC1 (19. 0%) and PC2 (8. 4%), which combined explained 27. 4% of variation in global expression. While the scree test indicated that PC3 and PC4 also explained a significant amount of variation (7. 0% and 6. 2%, respectively), there was no separation among treatments along those axes (S4 Fig). Download: PPTPowerPoint slidePNGlarger imageTIFForiginal imageFig 2. PCA biplot of first the two principal components, constructed from all expressed genes. gene expression was less variable among crabs reared in OA treatments compared to those reared in ambient conditions. Using a test of multivariate homogeneity of group dispersions, we found that variation in global gene expression differed by treatment (F(2,39) = 4. 9, p = 0. 012). Pairwise comparisons indicated that variation differed between ambient and severe OA (p-valuepermuted = 8. 3e-3), but did not differ between ambient and moderate OA (p-valuepermuted = 0. 32) or moderate and severe OA (p-valuepermuted = 0. 25). Co-expression network analysis We performed a weighted gene co-expression network analysis (WGCNA) to identify groups of genes that were co-expressed (i. e. gene modules), and for which expression correlated with pCO2 concentration. The 74,778 examined genes were assigned to 41 modules, 14 of which had eigengenes that correlated significantly with pCO2 (S2 Table). For eight of the modules, which contained in total 22,537 genes, the eigengenes correlated negatively with pCO2 indicating that expression decreased as OA treatment became more severe. Six modules, containing 19,248 genes in total, correlated positively with pCO2, indicating higher expression in OA treatments. Differential gene expression Analysis in DESeq2 identified 6,806 genes that were differentially expressed among treatments (9. 1% of all examined genes).

Output: contrast


Input: Claim: Curcumin, found in turmeric, has anti-inflammatory properties. [SEP] Text: After exposure, cells were detached from the flasks using 0. 05% trypsin and 0. 01% EDTA, centrifuged, washed by PBS and the pellet was suspended in 300 µl phosphate-buffered saline (PBS) and incubated with 0. 5 µl annexin V-FITC and 0. 5 µl of propidium iodide at room temperature for 30 min in the dark. Stained cells were subjected to flow cytometry analyses, using a “LSRII” or “CantoII” cytometer instrument from BD Biosciences. Data analysis was made by FACS Diva software v. 6. 1. 3. The experiments were repeated 5 independent times. percentage depending on the various phases of the cell cycle was measured by using the fluorescent DNA dye 7-aminoactinomycin D (7-AAD) (Sigma, USA), as previously described 29. Briefly, oral cancer cells were treated with different PAC concentrations (1 µM, 2. 5 µM, 5 µM or 10 µM) for 24 h. They were then incubated in the presence of 7-AAD at 1 µg/mL for 30 min at 37ºC. The fluorescence intensities of DNA were detected with “LSRII” or “CantoII” (BD Biosciences). Data were analyzed by FACSDiva software v. 6. 1. 3 and experiments were done in triplicates, n = 5. Pancaspase activity assay by flow cytometry The pancaspase activity in ca 9–22 treated by various concentration of PAC was evaluated by use Kit (FITC-VAD-FMK) from Calbiochem. Briefly; 1 × 106 cells were treated or not by different concentration of PAC for 24 h. After PAC-treatment, 1 µl FITC-VAD-FMK was added into each culture and incubate for 1 h in a 37 °C incubator with 5% CO2 and then, the cells were re-suspended in 0. 3 ml wash buffer before quantification by flow cytometry using the FL-1 channel (n = 5).

Output: pass


Input: Claim: The majority of research in graphene focuses on its application in supercapacitors and energy storage due to its high electrical conductivity. [SEP] Text: Description Three-Materials for -dimensional (3D) graphene architectures have allured remarkable attention for supercapacitor (SC) applications owing to their highly accessible surface area, low density, structural interconnectivity (micro-, meso- and macro-interconnected pores), excellent electrical conductivity and good mechanical strength. Overall supercapacitance performance of 3D graphene-based SCs has been due to enhanced accessibility of the electrode surface to electrolyte ions, which also provides conductive channels for electron transfer. In addition, 3D graphene structures provide an ideal template for active material decoration. In this book 10. 21741/9781644900550-4 Part of the book on Graphene as Material for Supercapacitors 23. [2] A. Chatzivasileiadi, E. Ampatzi, I. Knight, Characteristics of electrical energy storage technologies and their applications in buildings, Renew. Sustain. Energy Rev. 25 (2013) 814–830. [3] of Nanoballs for Supercapacitor, ACS Nano 7 (2013) 6047–6055. [4] K. Chatzivasileiadi, Large stationary batteries for deployment in grid-connected photovoltaic and other renewable energy power plants, 7th Conference and Exhibition, November 2012. [5] P. Simon, Y. Gogotsi, Materials for electrochmeical capacitors, Nat. Mater. (2008) 845–854. [6] R. M. A. S. Rajakaruna, Small signal transfer functions of the classical boost converter supplied by ultracapacitor banks, Second IEEE conference on Ind. Electron. and applications (2007) 692–697. [7] T. Wei, S. Wang, Z. Qi, A supercapacitor based ride-through system for industrial drive applications, Proc. 2007 IEEE Int. Conf. Mechatronics Autom. ICMA 2007. (2007) 3833–3837. [8] S. C. Smith, P. K. Sen, Ultracapacitors and energy storage: Applications in electrical power system, 40th North Am. Power Symp. NAPS2008. (2008) 1–6. [9] M. Vangari, T. Pryor, L.

Output: corroborate


Input: Claim: Ocean acidification only affects animals with hard shells, and fish are immune. [SEP] Text: The number of differentially expressed genes increased with OA severity: 1,459 genes differed between ambient and moderate OA (526 upregulated in moderate OA, 933 downregulated, Figs 3 & 4A), and 6,257 genes differed between ambient and severe OA (2,350 upregulated in severe OA, 3,907 downregulated, Figs 3 & 4B). Only 47 genes differed between moderate and severe OA treatments (22 upregulated in severe OA, 25 downregulated, Figs 3 & 4C), indicating that the two OA treatments induced a similar transcriptional response (S3 Table). There was high overlap between the differentially expressed genes and the co-expression network analysis (WGCNA), with 92. 8% of DEG’s assigned to one of the 14 pCO2-correlated co-expression modules. Download: PPTPowerPoint slidePNGlarger imageTIFForiginal imageFig 3. Venn diagram showing the number of differentially expressed genes among each pairwise OA treatment contrast, and the number that are shared among contrasts. Download: PPTPowerPoint slidePNGlarger imageTIFForiginal imageFig 4. Pairwise differential expression among OA treatments for (A) ambient (pH 8. 0) vs. moderate OA (pH 7. 8), (B) ambient vs. severe OA (pH 7. 5), and (C) moderate OA vs. severe OA. Volcano plots (left panels) show expression of all genes. Each point represents a unique gene, with non-black points residing above the dotted line representing differentially expressed genes. -Log10 p-value is along the y-axes, with higher numbers indicating higher significance, and log2 fold change is along the x-axes, with higher absolute values indicating larger differences among treatments. Differentially expressed genes (p-adj < 0. 05) are color-coded to indicate those that are expressed at higher levels in ambient treatment (blue), moderate OA (orange), or severe OA (red). Points with darker shades indicate those that have |Log2FC| >0. 5, which were used in functional analyses.

Output: contrast


Input: Claim: Curcumin, found in turmeric, has anti-inflammatory properties. [SEP] Text: Cell Autophagy: To determine the in vitro effect of PAC on Ca9-22 cells autophagy in, we used ICT’s (Red) from ImmunoChemistry Technologies. Briefly, Ca9-22 cells were stimulated or not with various concentrations of PAC for 24 h, detached from the culture plates, then incubated with a diluted Autophagy Probe, Red at 1:50 for 1 h in the dark, washed three times with the cellular assay buffer. Stained cells were afterwards analyzed by flow cytometry, (n = 4). Stress: Expression of cellular ROS was evaluated by using a specific assay kit from ImmunoChemistry Technologies. In short, Ca9-22 cells were seeded and cultured from 24 h, then were exposed to different concentrations (1 µM, 2. 5 µM, 5 µM or 10 µM) of PAC for 24 h. Total ROS Green dye molecule was added to 106 cells/mL of each culture condition for 1 h in the dark at 37 °C before making the analysis with BD Accuri C6 (BD Biosciences). For evaluating levels of intracellular GSH, ThioBright™ Green reagent was first reconstituted in DMSO, added to the cell suspension (1:200), and then incubated for 30 min in the dark. After washing with PBS twice, fluorescence intensity was determined by System (BD Biosciences), and the percentage of GSH-positive cells was calculated. Experiments were done in triplicates. Potential (MMP) MMP expression reduced by PAC was measured by DiOC(2)(3)-based flow cytometry. Ca9-22 cells (106) were seeded into a T325 flask. After overnight incubation for adherence, cells were treated with different concentrations of PAC (0, 1, 2. 5, 5 and 10 µM). After 24 h of treatment, 50 nM of DiOC2(3) were added to each condition for 30 min. Celles were collected and subjected to flow cytometry analyses using a “LSRII” or “CantoII” cytometer instruments from BD Biosciences. Data analysis was made by FACSDiva software v. 6. 1. 3. , (n = 4).

Output: pass


Input: Claim: The majority of research in graphene focuses on its application in supercapacitors and energy storage due to its high electrical conductivity. [SEP] Text: Jiang, Supercapacitors: Review of Materials and Fabrication Methods, J. Energy Eng. 139 (2013) 72–79. [10] Y. Wang, Supercapacitor devices based on graphene materials, J. Phys. Chem. C. 113 (2009) 13103–13107. [11] V. V. N. Obreja, Supercapacitors specialities-Materials review, AIP Conf. Proc. 1597 (2014) 98–120. [12] Z. S. Iro, C. Subramani, S. S. Dash, A brief review on electrode materials for supercapacitor, Int. J. Electrochem. Sci. 11 (2016) 10628–10643. [13] B. Francesco, C. Luigi, Y. Guihai, M. Stoller, V. Tozzini, C. F. Andrea, Graphene, related two-dimensional crystals, and hybrid systems for energy conversion and storage, Science 347 (2015) 27–43. [14] A. González, E. Goikolea, J. A. Barrena, R. Mysyk, Review on supercapacitors: Technologies and materials, Renew. Sustain. Energy Rev. 58 (2016) 1189–1206. [15] J. A. Fernández, T. Morishita, M. Toyoda, M. Inagaki, F. Stoeckli, T. A. Centeno, Performance of mesoporous carbons derived from poly(vinyl alcohol) in electrochemical capacitors, J. 175 (2008) 675–679. [16] D. Qu, Studies of the activated carbons used in double-layer supercapacitors,. 41 (2014) 1108–1110. [17] K. Nanaji, V. Upadhyayula, T. N. Rao, S. Anandan, Robust, environmentally benign synthesis of nanoporous graphene sheets from biowaste for ultrafast supercapacitor application, ACS Sustain. Chem. Eng. 7 (2019) 2516-2529. [18] Y. Wang, Y. Xia, Recent progress in supercapacitors: From materials design to system construction, Adv. Mater.

Output: corroborate


Input: Claim: Ocean acidification only affects animals with hard shells, and fish are immune. [SEP] Text: There were 81 enriched processes in genes down-regulated in severe-OA reared crab. Many processes that were enriched in moderate-OA reared crab were also enriched in genes downregulated in severe-OA reared crab (Fig 7). Enriched processes that were uniquely downregulated in severe-OA reared crab included translation (the most significantly enriched down regulated process, p-value = 8. 6e-14), and those related to DNA replication, microtubules, immune function, and aerobic respiration (Fig 7, see S5 Table for full list). Download: PPTPowerPoint slidePNGlarger imageTIFForiginal imageFig 7. Ontology biological processes in genes that were downregulated in moderate OA (pH 7. 8, left column) and severe OA (pH 7. 3, right column) compared to ambient conditions (pH 8. 0). Point size indicates the -Log10 transformed p-value (all p-values < 0. 05), such that larger points are more significantly enriched. Only the top 20 processes by p-value per OA treatment were included in this figure; see S5 Table for full list. biological processes were enriched in genes that were differentially expressed among crab reared in moderate vs. severe OA. Two processes involved in molecular chaperone activity via the heat shock complex were enriched in genes with increased expression in the moderate OA-reared crab, and three processes involved in transposition were enriched in genes with increased expression in the severe OA-reared crab (S5 Table). Enrichment analysis of genes potentially critical in an OA environment. Of the 526 and 2,350 genes that were upregulated in moderate and severe OA relative to ambient treatment, respectively, 66 (13%) and 1,319 (56%) were expressed at consistent levels across individuals within treatments (CV < 3%, hereafter referred to as low-variance genes). These upregulated low-variance genes are of interest as they may provide critical functions in the OA environment.

Output: contrast


Input: Claim: Curcumin, found in turmeric, has anti-inflammatory properties. [SEP] Text: Assessment of To evaluate the mitochondrial superoxide production following treatments with PAC, we used an indicator named MitoSOX Red (Molecular Probes, Invitrogen), according to the manufacturer’s instructions, and afterwards performed a flow cytometry analysis. PAC-treated cells (1 µ M, 2. 5 µM, 5 µM or 10 µM) were incubated for 30 min with 5 µM MitoSOX at 37ºC in 5% CO2. They were then resuspended in 1 mL PBS, and analyses by means of flow cytometry with “LSRII” or “CantoII” (BD Biosciences) (n = 3). The effect of PAC at 5 µM on various proteins involved in MAPK pathways, apoptosis, autophagy, and cell cycle was evaluated by making use of Western blots. Briefly, 106 cells (stimulated and unstimulated) were harvested for extraction by RIPA lysis buffer. Protein concentrations were determined by the Bradford assay. For each sample, a mixture of 20 µg to 60 µg was added to an equal amount of loading buffer containing 10% β-mercaptoethanol and migrated through a 7–15% acrylamide gel. After electrophoretic separation, proteins were transferred from the gel to a nitrocellulose membrane to be blocked in a 5% milk solution for 1 h at 24ºC and put overnight in an incubator with appropriate concentrations of antibodies. Finally, a band detection was performed by an ECL (), according to instructions from its manufacturer. Statistical analysis All our experiments were repeated more than three times and the statistical significance obtained between controls (untreated cells) and tests (treated) with various concentrations of PAC was expressed as mean ± SD. Differences between values were determined if statistically significant using a one-way ANOVA. A p-value < 0. 05 was defined as significant. Results PAC selectively inhibits the growth of oral cancer cells, and increases apoptosis To evaluate the effect of PAC on normal and gingival cells affected by cancer to proliferate, we used the MTT assay. As presented in Fig. 1, PAC at various concentrations decrease the growth of oral cancer cells. This decrease was more important with higher concentration of PAC. Indeed, at 10 µM oral cancer cell growth was almost drastically reduced.

Output: pass


Input: Claim: The majority of research in graphene focuses on its application in supercapacitors and energy storage due to its high electrical conductivity. [SEP] Text: 25 (2013) 5336–5342. [19] C. Liu, Z. Yu, D. Neff, A. Zhamu, B. Z. Jang, Graphene-based supercapacitor with an ultrahigh energy density, Nano Lett. 10 (2010) 4863–4868. [20] H. Pan, J. Li, Y. P. Feng, Carbon nanotubes for supercapacitor, Nanoscale Res. Lett. 5 (2010) 654–668. [21] H. I. Becker, Patent Office, (1940). [22] M. Dinari, M. M. Momeni, M. Goudarzirad, Nanocomposite films of polyaniline/graphene quantum dots and its supercapacitor properties, Surf. Eng. 32 (2016) 535–540. [23] S. Zhang, L. Sui, H. Dong, W. He, L. Dong, L. Yu, High-performance supercapacitor of graphene quantum dots with uniform sizes, ACS Appl. Mater. Interfaces 10 (2018) 12983–12991. [24] S. Mondal, U. Rana, S. Malik, Graphene quantum dot-doped polyaniline nanofiber as high performance supercapacitor electrode materials, Chem. Commun. 51 (2015) 12365–12368. [25] K. Lee, H. Lee, Y. Shin, Y. Yoon, D. Kim, H. Lee, Highly transparent and flexible supercapacitors using graphene-graphene quantum dots chelate, 26 (2016) 746–754. [26] Q. Chen, Y. Hu, C. Hu, H. Cheng, Z. Zhang, H. Shao, L. Qu, Graphene quantum dots-three-dimensional graphene composites for high-performance supercapacitors, Phys. Chem. Chem. Phys. 16 (2014) 19307–19313. [27] Y. Hu, Y. Zhao, G. Lu, N. Chen, Z. Zhang, H. Li, H. Shao, L.

Output: corroborate


Input: Claim: Ocean acidification only affects animals with hard shells, and fish are immune. [SEP] Text: Low-variance genes upregulated in severe OA treatment were enriched for biological processes involved in transcription regulation, DNA integration (transposon activity), nervous system processes (neurogenesis, signal transduction, calcium ion-regulated exocytosis of neurotransmitter), developmental processes (multicellular organism development, keratinocyte differentiation, glycosaminoglycan biosynthesis), regulation of cardiac muscle cell contraction, and potassium ion transport (S6 Table). Low-variance genes upregulated in moderate OA treatment were enriched for chromatin organization and transposition (S6 Table). In contrast, low-variance genes that were upregulated in ambient treatment relative to either OA treatment were primarily enriched for processes involved in protein biosynthesis, glycolysis, and ion transport (including hydrogen ion transport) (S6 Table). Transposable element composition. A large portion of the red king crab transcriptome mapped to transposable elements (20,860 TEs), comprising 28% of the 74,778 analyzed genes, and 64% of the 32,435 annotated genes. These TEs mapped to 67 distinct IDs, all of which were retroelements (S7 Table). Transposable element activity was high in all treatments, but activity increased with OA severity. Of the annotated genes that were upregulated in severe OA compared to ambient and moderate OA treatments, 65% and 86% mapped to TE’s, respectively, while 51% of genes upregulated in moderate OA compared to ambient were TE’s (Fig 8, S8 Table). In contrast, 31% and 28% of genes that were more abundant in ambient treatment compared to moderate and severe OA mapped to TEs (Fig 8, S8 Table). A large percentage of upregulated low-variance genes were also transposable elements, comprising 50% and 60% of those genes in moderate and severe OA-treated crab, respectively. In contrast, 16% of the low-variance upregulated genes in ambient-pH-reared crab were transposable elements (S8 Table). The TE composition of co-expressed gene modules was similar- on average the percent of genes that increased and decreased with pCO2 was 73% and 51%, respectively (Fig 8).

Output: contrast


Input: Claim: Curcumin, found in turmeric, has anti-inflammatory properties. [SEP] Text: Interesting, the decrease of normal human gingival epithelial cells growth following the exposure to 10 uM of PAC was too low compared to cancer cells. With 5 µM of PAC, there was an 80% growth reduction of oral cancer cells, but only 10–20% growth reduction of primary human gingival epithelial cells. These results demonstrated that normal human cells exhibit great resistance to PAC, in comparison to Ca9-22 cells. The effect of PAC on cancer cells was observable even at low concentration with an LC50 is around 3 µM. Its proliferation level decreased, from 86. 21% ± 4. 03% (p = 0. 007), 78. 41% ± 8. 23% (p = 0. 02), 24. 37% ± 7. 31% (p = 2. 39 × 10− 5) to 9. 85% ± 2. 16% (p = 6. 46 × 10− 9) respectively with PAC concentrations of 1, 2. 5, 5 or 10 µM (Fig. 1A). With GEC cells, a slight decrease was observed only at the highest concentration of PAC (10 µM) after 48 h (p = 0. 02) and 72 h (p = 0. 03), but any significant inhibition for all treatments (1 µM to 10 µM) was noticeable after 24 h. (Fig. 1B). These outcomes were confirmed using a cytotoxicity assay (Fig. 1C and 1D), a treatment with PAC was inducing dose- and time-dependent cell death in Ca9-22 (Fig. 1C) but not in GEC cells (Fig. 1D), suggesting a selective toxic effect of PAC on cancer rather than normal gingival epithelial cells. We also demonstrated a significant effect of PAC reducing the capacity of oral cancer cells to form colony but not in GEC (Fig. 1E). These outcomes clearly indicate as well that PAC is having preferentially anti-proliferative activity and cytotoxicity against Ca9-22 cells, but not on normal human gingival epithelial cells. We demonstrated that anti-proliferative effect of PAC on oral cancer cells goes through a cell cycle modification.

Output: pass


Input: Claim: The majority of research in graphene focuses on its application in supercapacitors and energy storage due to its high electrical conductivity. [SEP] Text: Qu, Graphene quantum dots–carbon nanotube hybrid arrays for supercapacitors, Nanotechnology 24 (2013) 1-7. [28] A. B. Ganganboina, A. Dutta Chowdhury, R. A. Doong, New avenue for appendage of graphene quantum dots on halloysite nanotubes as anode materials for high performance supercapacitors, ACS Sustain. Chem. Eng. 5 (2017) 4930–4940. [29] H. Jia, Y. Cai, J. Lin, H. Liang, J. Qi, J. Cao, J. Feng, W. D. Fei, Heterostructural graphene quantum dot/MnO2 nanosheets toward high-potential window electrodes for high-performance supercapacitors, Adv. Sci. 5 (2018) 1–10. [30] P. Dhar, S. S. Gaur, A. Kumar, V. Katiyar, Cellulose nanocrystal templated graphene nanoscrolls for high performance supercapacitors and hydrogen storage: An experimental and molecular simulation study, Sci. Rep. 8 (2018) 1–15. [31] Z. Xu, B. Zheng, J. Chen, C. Gao, Highly efficient synthesis of neat graphene nanoscrolls from graphene oxide by well-controlled lyophilization, Chem. Mater. 26 (2014) 6811–6818. [32] F. Zeng, Y. Kuang, Y. Wang, Z. Huang, C. Fu, H. Zhou, Facile preparation of high-quality graphene scrolls from graphite oxide by a microexplosion method, Adv. Mater. 23 (2011) 4929–4932. [33] and morphology tuning of graphene oxide nanoscrolls, Nanoscale. 8 (2016) 6783–6791. [34] B. Zheng, Z. Xu, C. Gao, Mass production of graphene nanoscrolls and their application in high rate performance supercapacitors, Nanoscale. 8 (2016) 1413–1420. [35] K. Mohanapriya, N.

Output: corroborate


Input: Claim: Ocean acidification only affects animals with hard shells, and fish are immune. [SEP] Text: A likelihood ratio test examined the proportions of gene sets that were TEs, and found higher TE proportions in upregulated gene sets compared to downregulated gene sets (χ2 (2, N = 20) = 10. 7, p = 0. 001, Fig 8). Download: PPTPowerPoint slidePNGlarger imageTIFForiginal imageFig 8. The percent of genes in each co-expressed gene module and differentially expressed gene set (DEGs) that were transposable elements (TEs), categorized by whether genes were downregulated or upregulated in OA relative to ambient treatment, which shows the high percentage of TEs in OA-upregulated genes. The proportion of all genes examined that were TEs (64%) is indicated by the dotted line. TEs were identified by searching within the protein names for the words transposon, transposable, LINE, retrotransposable element, transposable element, mobile element jockey, and pol polyprotein. Lines in the middle of each boxplot indicate the median percent TEs in downregulated and upregulated gene sets. DiscussionCalcifying marine species living at high latitudes may be particularly vulnerable to the effects of OA due to more extreme changes projected to occur in those regions [1, 15]. Crustaceans in some high latitude regions are likely already experiencing acidified conditions seasonally; at present the shelf drops to seasonal lows of around pH 7. 5 [13] and mean bottom pH is projected to drop by a further 0. 3 units by 2100 [15]. Red king crab, which is one of several valuable commercial fisheries in Alaskan waters, are sensitive to changes in ocean chemistry at the juvenile stage, resulting in high mortality and decreased growth [51, 86]. However, our supporting study [55] found that red king crab are surprisingly resilient to OA when exposure begins early in life—at hatch—with no impact to survival, growth, or development through the early juvenile (C1) stage. Here, we explore aspects of red king crab molecular physiology that are altered by long term exposure to acidification, and which may be critical to their survival, to understand their adaptive potential and improve population predictions. We provide the first study to describe the molecular signatures of juvenile red king crab that were reared from the larval stage in acidified conditions.

Output: contrast


Input: Claim: Curcumin, found in turmeric, has anti-inflammatory properties. [SEP] Text: Indeed, Ca9-22 cells treated with 5 µM showed a cell-cycle arrest, by increasing the expression of cyclin-dependent kinase inhibitors (CDKIs) such as p21, p27, p16 or p53 and Rb (retinoblastoma protein) which are tumor suppressor genes known to be inactive in most cancer types, while it inhibits oncogenes such as cyclin D1 or c-Myc. In addition, PAC can to reduce DNA damage on oral cancer cells by inhibition of H2A. X protein expression (Fig. 2A). The effect of PAC on cancer cells is involving specific inhibition of cell proliferation signaling pathways. Indeed, we demonstrated a significant decrease of ERK1/2 phosphorylation, p38, STAT and AKT phosphorylation. Also, PAC at 5 µM was decreasing the expression of NF-kB and Wnt/β-catenin signaling pathways (Fig. 2B). Effect of treatments with PAC on cell cycle distribution in oral cells Cell cycle distribution, on Ca9-22 PAC-treated cells, was evaluated by flow cytometry using the fluorescent DNA dye 7-aminoactinomycin D (7-AAD). Briefly, these were under treatment with different PAC concentrations for 24 h. Then, they were responding to PBS buffer before being incubated with 7-AAD Fig. 3 is presenting that different concentrations of PAC were altering Ca9-22 cell cycle distribution. However, percentage of G0/G1 population decreases dramatically, from 85. 95% ± 7. 14% in untreated cells to 9. 9% ± 3. 67% in cells treated with 10 µM of PAC. In contrary, the S population increases from 6. 85% ± 3. 74% in controls to 38. 95% ± 17. 18% using the same concentration it. The G2/M population is also doing it as the previous one, from 5. 05% ± 1. 34–48. 7% ± 13. 85% with the same treatment (Fig. 3A and 3B). PAC induces apoptosis in oral cancer cells by targeting the intrinsic mitochondrial pathway. In determining the type of death induced by various concentrations of PAC on oral cancer cells, the fluorochrome-labeled annexin V with PI was used.

Output: pass


Input: Claim: The majority of research in graphene focuses on its application in supercapacitors and energy storage due to its high electrical conductivity. [SEP] Text: Jha, Fabrication of one dimensional graphene nanoscrolls for high performance supercapacitor application, Appl. Surf. Sci. 449 (2018) 461–467. [36] M. Yan, F. Wang, C. Han, X. Ma, X. Xu, Q. An, L. Xu, C. Niu, Y. Zhao, X. Tian, P. Hu, H. Wu, L. Mai, Nanowire templated semihollow bicontinuous graphene scrolls: Designed construction, mechanism, and enhanced energy storage performance, J. Am. Chem. Soc. 135 (2013) 18176–18182. [37] B. N. Zheng, C. Gao, Preparation of graphene nanoscroll/polyaniline composites and their use in high performance supercapacitors, Mater. 31 (2016) 315–320. [38] H. Cheng, C. Hu, Y. Zhao, L. Qu, Graphene fiber: A new material platform for unique applications, NPG Asia Mater. 6 (2014) e113-13. [39] Z. Xu, C. Gao, Graphene fiber: A new trend in carbon fibers, Mater. Today. 18 (2015) 480–492. [40] S. H. Aboutalebi, R. Jalili, D. Esrafilzadeh, M. Salari, Z. Gholamvand, S. Aminorroaya Yamini, K. Konstantinov, R. L. Shepherd, J. -performance multifunctional Graphene yarns: Toward wearable all-carbon energy storage textiles, ACS Nano 8 (2014) 2456–2466. [41] W. Cai, T. Lai, J. Lai, H. Xie, L. Ouyang, J. Ye, C. Yu, Transition metal sulfides grown on graphene fibers for wearable asymmetric supercapacitors with high volumetric capacitance and high energy density, Sci. Rep. 6 (2016) 1–9. [42] Q. Chen, Y. Meng, C. Hu, Y. Zhao, H. Shao, N. Chen, L.

Output: corroborate


Input: Claim: Ocean acidification only affects animals with hard shells, and fish are immune. [SEP] Text: Expression of nearly half of all genes (44%) correlated significantly with pCO2, suggesting a strong molecular response to OA conditions, contrary to the phenotypic results. There was a general reduction in transcriptional activity and inter-individual variability in OA-reared crab, suggesting that OA reduces the breadth of physiological functions compared to crabs reared in ambient seawater. Functional analysis of downregulated genes indicates that energy production is depressed in OA conditions, which is likely associated with decreases in biosynthesis, the immune system, and myriad metabolic processes. Downregulated lipid and carbohydrate metabolic processes suggest a shift in metabolic strategy to protein catabolism, possibly to reduce CO2 production and facilitate acid/base regulation. The limited energy is shunted towards transcriptional regulation mechanisms, signaling systems, and control of growth. Transposable elements (TEs) in OA-reared crab were highly active, particularly in the most severe OA treatment, and were expressed at consistent levels, suggesting that TEs play a role in the OA-response. In the remainder of this section we describe processes that are suppressed and triggered by OA exposure during development in red king crab, and expand on effects to TE activity. Finally, we discuss differences in gene expression variation observed among treatments, and consider whether there were significant genetic differences among treatment groups that may have contributed to the observed gene expression profiles. Processes that are suppressed in OA-reared crab OA resulted in a widespread downregulation of metabolic processes involved in multiple respiratory pathways, metabolism of a variety of compounds, and protein synthesis machinery (Fig 7). Reduced metabolic activity may be one way that red king crabs mitigate acidosis [87, 88]. Carbon dioxide is produced during respiration, therefore a decrease in metabolic activity reduces internal CO2 production. Decreased mitochondrial activity can also reduce oxidative cellular stress by reducing mitochondrial production of reactive oxygen species, which might otherwise increase due to environmental stressors [89–91]. Another outcome of changes in metabolic activity could be a shift in metabolic strategy away from carbohydrate and lipid substrates and towards proteins to improve acid/base regulation [92, 93]. Protein catabolism is less energy efficient, but instead of CO2 it produces ammonia and bicarbonate, which can be used to buffer intracellular pH [94].

Output: contrast


Input: Claim: Curcumin, found in turmeric, has anti-inflammatory properties. [SEP] Text: Our results show the presence of 4 different sub-populations referring to live cells (annexin V-/PI-), early apoptotic cells (annexin V+/PI-), late apoptotic cells (annexin V+/PI+) and necrotic cells (annexin V-/PI+). After 24 h of exposure to PAC, the percentage for the first population decreases from 96. 48% for untreated to 71. 29% with 5 µM, and 18. 32% with 10 µM. Conversely, the early apoptotic cells were increasing from 2. 56% with the control to 18. 62 with 5 µM. In late apoptotic population was much higher with PAC exposure ranging from, 1. 09% with the controls to 55. 46% with 10 µM of PAC. The necrotic population was also increasing from 0. 87% with the untreated cells to 24. 48% with 10 µM PAC treated oral cancer cells (Fig. 4A). To confirm apoptosis –inducing ability of PAC concentration on oral cancer cells, we evaluated caspase activation by using flow cytometry. Our results show that PAC dose – dependant increased apoptosis in terms of percentage of positive pancaspase in Ca9-22 cells (Fig. 4B). The effect of PAC of caspase signaling for oral cancer cell apoptosis was confirmed by Western blotting (Fig. 4C) showing that PAC at 5 µM strongly decreases Bcl-2 expression and increases that of pro-apoptotic Bax and cytochrome C as well as allows cleavage of PARP-1. The effect of PAC promoting cancer cell death involves caspase 3/9 (Fig. 4C). PAC promotes autophagy in oral cancer cells by targeting LC3B and p62 protein expression. To determine the different cellular mechanisms by which PAC exerts its anti-oral cancer function, we investigated the effect of PAC on cell autophagy. As shown in Fig. 5A and 5B, PAC at 5 µM strongly leads to autophagy in cancer cells. The percentage of positive cells detected by the (Red) was increasing from 0. 1125% ± 0. 15% in controls to 10. 57% ± 3. 35% (p = 0. 00066) and 28. 59 ± 13. 78% (p = 0.

Output: pass


Input: Claim: The majority of research in graphene focuses on its application in supercapacitors and energy storage due to its high electrical conductivity. [SEP] Text: Qu, MnO2-modified hierarchical graphene fiber electrochemical supercapacitor, J. 247 (2014) 32–39. [43] K. Gopalsamy, Z. Xu, B. Zheng, T. Huang, L. Kou, X. Zhao, C. Gao, Bismuth oxide nanotubes-graphene fiber-based flexible supercapacitors, Nanoscale 6 (2014) 8595–8600. [44] L. Liu, Y. Yu, C. Yan, K. Li, Z. Zheng, Wearable energy-dense and power-dense supercapacitor yarns enabled by scalable graphene-metallic textile composite electrodes, Nat. Commun. 6 (2015) 1–9. [45] Q. Yang, Z. Xu, C. Gao, Graphene fiber based supercapacitors: Strategies and perspective toward high performances, J. Energy Chem. 27 (2018) 6–11. [46] M. D. Stoller, S. Park, Y. Zhu, J. An, R. S. Ruoff, Graphene-based ultracapacitors, Nano Lett. 8 (2008) 6–10. [47] L. L. Zhang, X. Zhao, M. D. Stoller, Y. Zhu, H. Ji, S. Murali, Y. Wu, S. Perales, B. Clevenger, R. S. Ruoff, Highly conductive and porous activated reduced graphene oxide films for high-power supercapacitors, Nano Lett. 12 (2012) 1806-1812. [48] G. Sun, J. Liu, X. Zhang, X. Wang, H. Li, Y. Yu, W. Huang, H. Zhang, P. Chen, Fabrication of ultralong hybrid microfibers from nanosheets of reduced graphene oxide and transition-metal dichalcogenides and their application as supercapacitors, Angew. Chemie Int. Ed. 53 (2014) 12576–12580. [49] Q. Li, X. Guo, Y. Zhang, W. Zhang, C. Ge, L. Zhao, X. Wang, H.

Output: corroborate


Input: Claim: Ocean acidification only affects animals with hard shells, and fish are immune. [SEP] Text: While we did detect downregulation of carbohydrate & lipid metabolism that suggests a metabolic shift (Fig 7), and increased transmembrane signaling, we did not detect increased ion regulation processes in response to OA. It is possible that ion transport was indeed higher, but was uncoupled from mRNA levels in our fully OA-acclimated crab [95]. Pairing gene expression with more direct physiological assays (e. g. excreted O:N ratio, in vivo Na+,K+-ATPase activity) would improve interpretation of how red king crab metabolic and ion regulation strategies respond to OA. Metabolic changes are quite common in response to OA [96, 97]. OA can cause metabolic depression (and more generally, reduction in gene activity) in crustaceans and other marine invertebrates [98–102]. In the present study, metabolic processes were downregulated in both the moderate and severe OA treatments, which likely reflects the sensitivity of red king crab to changes in ocean chemistry. Given these findings, we might expect metabolic rate to decrease in response to OA in red king crab. In previous respiration trials [50] there was no change to metabolic rate measured by oxygen consumption in juvenile red king crab exposed to the same OA conditions after three weeks. The decreased metabolic gene activity in the present study could reflect an acclimatory response that is only induce by long-term and/or multi-stage exposure—in our case three months from hatching—which may influence the physiology of crabs in a way that short term- or single life stage- exposure does not [97]. Decreased expression of metabolic functions may also reflect an interactive effect between OA treatment and seasonal temperature increase, which was not a factor in Long et al. [50] (they held crab at 5°C). Future studies should pair gene expression analysis with more direct measurements of metabolic rate (and other traditional physiological measurements) at varying temperatures to untangle the effects of OA and temperature on red king crab metabolic functioning. OA affected the red king crab immune system. Genes involved in neuroinflammation (microglial cell activation), viral response, and the innate immune system decreased with pCO2, although effects were less pronounced compared to metabolic changes. OA can negatively affect the immune system in crustaceans [22, 35, 103–105] and other marine invertebrates [106–108].

Output: contrast


Input: Claim: Curcumin, found in turmeric, has anti-inflammatory properties. [SEP] Text: 0030) respectively for 5 µM or 10 µM of PAC. These observations were confirmed by showing an overexpression (high protein levels) of two autophagy markers (LC3B and p62) in Ca9-22 cells (Fig. 5C). PAC suppresses the process of epithelial-to-mesenchymal transition in oral cancer cells and mediates the anti-metastatic activity In Fig. 6A, we demonstrated that PAC at 5 µM inhibits the EMT in oral cancer cells, as shown by the increase of E-cadherin, while vimentin expression was decreasing. Metastatic cancer is also closely associated with cell migration. In this sense, we investigated the effect of various PAC concentrations on Ca9-22 cells migrating under treatment by using the (Scratch) Assay. As shown Fig. 6B, PAC at various concentrations was able to decrease the migration of oral cancer cells. Indeed, after 6 h of treatments with PAC, 100% of the scratch area was covered due to cell migration, in the controls, while less than 47% and 33% of the scratched area being covered following exposure to 5 µM or 10 µM, respectively. ROS production during treatments with PAC in oral cancer cells. Our data shows that the intracellular ROS accumulation induced by the treatment for 24 h with PAC was evaluated using a flow cytometer (Fig. 7). The percentage of ROS (+) is low with increased concentration of PAC as treatment. By pre-treating with NAC, the role of PAC in reducing oxidative stress was confirmed. We, therefore, suspect that PAC acts as a powerful antioxidant by producing intracellular glutathione (GSH). GSH generation during treatments with PAC in oral cancer cells. Figure 8 shows that PAC strongly increase the GSH expression with 2. 5 µM compared to controls. The percentage of GSH positive is going up, from 41% with non-treated cells to 69% and 87%, respectively after cell exposure to 5 µM or 10 µM of PAC, for 24 h. PAC is then confirmed to play a crucial role in producing GSH, an antioxidant. Mitochondrial membrane potential is reduced during treatments with PAC in oral cancer cells. The MMP expression that is reduced by PAC was measured by the MitoProbe DiOC2(3) for Flow Cytometry. As shown in Fig.

Output: pass


Input: Claim: The majority of research in graphene focuses on its application in supercapacitors and energy storage due to its high electrical conductivity. [SEP] Text: Zhang, J. Chen, Z. Wang, L. Sun, Porous graphene paper for supercapacitor applications, J. Mater. Sci. Technol. 33 (2017) 793–799. [50] S. Gan, L. Zhong, T. Wu, D. Han, J. Zhang, J. Ulstrup, Q. Chi, L. Niu, Spontaneous and fast growth of large-area graphene nanofilms facilitated by oil/water interfaces, Adv. Mater. 24 (2012) 3958–3964. [51] G. Wallace, R. B. Kaner, M. Muller, S. Gilje, D. Li, Processable aqueous dispersions of graphene nanosheets, Nat. Nanotechnol. 3 (2008) 101–105. [52] F. Gu, H. Shin, C. -by-layer doping of few-layer graphene film, ACS Nano 4 (2010) 4595–4600. 10. 1021/nn1008808 [53] A. Davies, P. Audette, B. Farrow, F. Hassan, Z. Chen, J. Y. Choi, A. Yu, Graphene-based flexible supercapacitors: Pulse-electropolymerization of polypyrrole on free-standing graphene films, J. Phys. Chem. C. 115 (2011) 17612–17620. [54] S. Bose, T. Kuila, A. K. Mishra, R. -based nanostructured materials and their composites as supercapacitor electrodes, J. Mater. Chem. 22 (2012) 767–784. [55] -dimensional, one-dimensional, two-dimensional and three-dimensional nanostructured materials for advanced electrochemical energy devices, Prog. Mater. Sci. 57 (2012) 724–803. [56] H. Jiang, P. S. Lee, C. Li, 3D Carbon based nanostructures for advanced supercapacitors, Energy Environ. Sci. 6 (2013) 41–53. [57] L. Liu, Z. Niu, J.

Output: corroborate


Input: Claim: Ocean acidification only affects animals with hard shells, and fish are immune. [SEP] Text: The mechanisms by which OA alters immune function are not fully understood, and could relate to energy constraints and malfunction of immune-related enzymes and signaling/recognition pathways (among other mechanisms) [106, 108]. We find evidence that immunosuppression may in part be due to activation of the stress response [109]. Genes coding for the octopamine receptors and a receptor for tyramine, the direct precursor of octopamine, were upregulated in OA. Octopamine is the invertebrate orthologue to norepinephrine, and is one of the key neurotransmitters that regulates the acute stress-response [110]. Interestingly, molecular chaperones (HSP70 and DNAJA1), which respond to a variety of stressors by refolding or facilitating the destruction of damaged proteins [111, 112], increased in moderate OA but decreased substantially in severe OA, which suggests that those processes become ineffective as OA severity worsens. Further, it suggests that at moderate OA levels increased oxidative stress damages proteins, or that altered intracellular or extracellular pH affects folding patterns of proteins. While these changes did not ultimately result in mortality in the present study, our findings indicate that red king crab may be more vulnerable to co-occurring secondary stressors, such as thermal stress [53, 54], due to immunosuppression and changes in cellular stress-response mechanisms. Additional multi-stressor studies are needed that expose red king crab to a range of pH levels alongside other stressors, particularly pathogen challenges. Transposable elements are highly active in OA-reared crab Many of the genes upregulated under OA conditions were transposable elements (TEs, or transposons). While TEs were present in all gene sets, including those that were more active in ambient conditions, they comprised a much larger percentage of genes upregulated in OA-reared crab (Fig 8, S8 Table). TEs, or “jumping genes”, are DNA elements that move to new locations in the genome when activated, resulting in insertional mutations [113]. Retrotransposons, the class of TEs detected in our study, are mobilized by a copy-and-paste mechanism where the DNA sequence is transcribed and an RNA intermediary is then reverse-transcribed into a cDNA copy before being integrated into the genome [114].

Output: contrast


Input: Claim: Curcumin, found in turmeric, has anti-inflammatory properties. [SEP] Text: (2011) showing that PAC has similar effect against breast cancer 21,24. PAC decrease the cell proliferation through the deregulation of the cell cycle. Indeed, PAC treated cells showed high expression of CDKIs including p21, p27, p16 or p53 and Rb. It is well known that these genes suppress tumors, known for being inactive in most types of cancer and also for inhibiting genes with oncogenic functions like cyclin D1 or c-Myc. These molecules have a history of playing a major role in the cell-cycle arrest at G2/M, and ultimately induce inhibition of oral cancer cell proliferation. This is mediated by inhibiting several signaling pathways involved in cancer such as MAPK, Wnt and NF-kB. These data are in accordance with previous studies conducted by Al-Hujaily et al. (2011) and Al-Howail et al. (2016) reporting that PAC could be considered as a powerful nontoxic new chemotherapeutic agent against ER-negative tumors by triggering breast cancer cell apoptosis more than curcumin 21,24, downregulating ERα, c-Myc, cyclin D124 and inducing cell-cycle arrest at G2/M of this cancer cells 21. Also, our results demonstrate that PAC strongly induces oral cancer cell apoptosis mediated via the mitochondrial pathway by increasing the Bax/Bcl-2 protein expression ratios, activating caspase-3/9 and cleaving PARP-1. We conclude that the PAC anti-oral cancer properties were controlled by impairing pro-survival signaling pathways and activating apoptosis pathways such as ERK1/2 activation, but not MEK1/2, AKT activation, NF-kB 35; 36;37 and β-catenin,38 known to be implicated in tumor progression and as well for their resistance to chemo- and radiotherapy in several cancer types. Indeed, it is clearly reported that several oncoproteins such as cyclin D1 and p21 are strongly destabilized by inhibition of ERK1/2 activation. Wnt/β-catenin is the key oncogenic signaling pathway found to be highly active in various types of cancer, including oral cancer 38,39. Similar observation was reported on breast cancer cells being treated with PAC 21;24.

Output: pass


Input: Claim: The majority of research in graphene focuses on its application in supercapacitors and energy storage due to its high electrical conductivity. [SEP] Text: Chen, Flexible supercapacitors based on carbon nanomaterials, Chinese Chem. Lett. 29 (2018) 571–581. [58] Y. Ma, Y. Chen, Three-dimensional graphene networks : Synthesis, properties and applications, Natl. Sci. Rev. 2 (2015) 40-53. [59] X. Cao, Z. Yin, H. Zhang, Three-dimensional graphene materials: Preparation, structures and application in supercapacitors, Energy Environ. Sci. 7 (2014) 1850–1865. [60] Z. Chen, W. Ren, L. Gao, B. Liu, S. Pei, H. M. Cheng, Three-dimensional flexible and conductive interconnected graphene networks grown by chemical vapour deposition, Nat. Mater. 10 (2011) 424–428. [61] J. Sha, C. Gao, S. K. Lee, Y. Li, N. Zhao, J. M. Tour, Preparation of three-dimensional graphene foams using powder metallurgy templates, ACS Nano 10 (2016) 1411–1416. [62] -dimensional graphene and their integrated electrodes, 9 (2014) 785–807. [63] Q. Fang, Y. Shen, B. Chen, Synthesis, decoration and properties of three-dimensional graphene-based macrostructures: A review, Chem. Eng. J. 264 (2015) 753–771. [64] Z. Yang, S. Chabi, Y. Xia, Y. Zhu, Preparation of 3D graphene-based architectures and their applications in supercapacitors, Prog. Nat. Sci. Mater. Int. 25 (2015) 554–562. [65] Y. Ping, Y. Gong, Q. Fu, C. Pan, Preparation of three-dimensional graphene foam for high performance supercapacitors, Prog. Nat. Sci. Mater. Int. 27 (2017) 177–181. [66] H. Huang, Y. Tang, L. Xu, S. Tang, Y.

Output: corroborate


Input: Claim: Ocean acidification only affects animals with hard shells, and fish are immune. [SEP] Text: Increased transposable element activity in response to environmental stress is well documented in a variety of eukaryotes [115]. There have been, however, only a few studies to detect stress-activated TEs in marine crustaceans [116], and to our knowledge this is the first to do so in response to OA. While TEs comprise large portions of the genomes of arthropods [117], including crustaceans [65, 118], previous transcriptional characterization of stress-responses in crustaceans may have overlooked TE’s due to the common practice of masking repeat elements from genomes during analysis [119]. Recent reports from a wide range of other taxa have also implicated TEs in the OA-response, including anemones [120], clams [121], and diatoms [122]. This breadth of taxa, which now includes crustaceans, suggests that TEs are a common response to acidification exposure. Why TEs can become activated under stress is still under debate, as is whether they are detrimental or beneficial to an organism’s survival [114]. TEs may become more active because the mechanisms that suppress them are no longer functioning, which could negatively affect fitness due to pathological mutations. Alternatively, stress-activated TE’s could act as regulatory elements and/or increase diversity of proteins available to the organism or its descendants, which may cause beneficial phenotypic variation. TEs may also be a component of the antiviral defense system, as they can stimulate the antiviral inflammatory response [123, 124]. Deciphering why TE were more active in OA-reared crab is beyond the scope of this study. However, given that red king crab early life stages tolerated OA in this study, and that many genes that were upregulated at consistent levels among OA-exposed individuals were TEs, TE mobilization may be an effective component of the red king crab stress-response system, rather than simply the result of genomic instability. TEs are also strong facilitators of adaptive evolution [125, 126], thus OA-induced TE activity may benefit red king crab at an evolutionary scale, given that it could provide a mechanism for rapidly increasing genetic diversity [115, 127].

Output: contrast


Input: Claim: Curcumin, found in turmeric, has anti-inflammatory properties. [SEP] Text: Therefore, the strong PAC-dependent apoptosis could be of great value for inhibiting of oral cancer cell proliferation and inducing their death. In a recent study carried out by Al-Qasem et al. (2016), they have published the same effect of PAC on colon cancer cells. They concluded that PAC exhibits anti-colon cancer properties that have potential, by targeting cyclin D1 and suppressing JAK2/STAT3, AKT/mTOR and MEK/ERK signaling pathways as well as their common downstream effector which is cyclin D1 25. We suggest that PAC suppresses the activation of MAPK and the induction of caspase cleavage, leading to the downregulation of several MAPK targets including cyclin D1, p53, and caspases, as Bax, Bcl-2 and cytochrome C. All these are playing key roles in oral tumorigenesis 40–42. We demonstrated that PAC promoted oral cancer cell autophagy and reduced oxidative stress via ROS production. Our results obtained here are in disagreement with several current research studies focusing on the use of anticancer treatments having an effect while apoptosis in cancer cells is induced by increased production of reactive oxygen species (ROS). However, our findings are also in concordance with others recent studies reporting that increase in cellular O2 − and H2O2 by NADPH oxidase 43 are leading the stimulation of cancer cell proliferation by ROS-sensitive AKT/ERK signaling pathways and the reduction of cancer cell growth by blocking this enzyme 44; Dikalova et al. , 2010) by using mitoTEMPO, they significantly reduced ROS in cells and inhibited NADPH oxidase activity 43. As for Nazarewicz et al. (2013), they also reported that mitochondrial ROS scavenging by mitoTEMPO can block ROS-sensitive signaling pathways in cancer cells, cancer metabolism and induce cell death 45. Our data suggests that diminishing ROS production, by PAC-treated oral cancer cells, is altering cell signaling mediated by ROS-sensitive 1/2, and p38 as well as inducing cell death. The PAC-attenuation effect from these signaling pathways was the principal factor leading to dramatic changes in oral cancer cell metabolism.

Output: pass


Input: Claim: The majority of research in graphene focuses on its application in supercapacitors and energy storage due to its high electrical conductivity. [SEP] Text: Du, Direct formation of reduced graphene oxide and 3D lightweight nickel network composite foam by hydrohalic acids and its application for high-performance supercapacitors, ACS Appl. Mater. Interfaces 6 (2014) 10248–10257. [67] H. Huang, L. Xu, Y. Tang, S. Tang, Y. Du, Facile synthesis of nickel network supported three-dimensional graphene gel as a lightweight and binder-free electrode for high rate performance supercapacitor application, Nanoscale 6 (2014) 2426–2433. [68] J. Yang, E. Zhang, X. Li, Y. Yu, J. Qu, Z. Yu, Direct reduction of graphene oxide by ni foam as a high-capacitance supercapacitor electrode, ACS Appl. Mater. Interfaces 8 (2016) 2297-2305. [69] S. Sivaprakash, P. Sivaprakash, A facile synthesis of graphene foam as electrode material for supercapacitor, Mater. Res. Express 3 (2016) 1–7. [70] P. Miao, J. He, Z. Sang, F. Zhang, J. Guo, D. Su, X. Yan, X. Li, H. Ji, Hydrothermal growth of 3D graphene on nickel foam as a substrate of nickel-cobalt-sulfur for high-performance supercapacitors, J. Alloys Compd. 732 (2018) 613–623. [71] Z. Niu, J. Chen, H. H. Hng, J. Ma, X. Chen, A leavening strategy to prepare reduced graphene oxide foams, Adv. Mater. 24 (2012) 4144–4150. [72] W. Y. Tsai, R. Lin, S. Murali, L. Li. Gogotsi, P. Simon, Outstanding performance of activated graphene based supercapacitors in ionic liquid electrolyte from -50 to 80°C, 2 (2013) 403–411. [73] Y. Zhu, S. . Cai, P. J. Ferreira, A. . Thommes, D.

Output: corroborate


Input: Claim: Ocean acidification only affects animals with hard shells, and fish are immune. [SEP] Text: It would be informative to examine whether the stress-induced mobilization of TE’s increases mutation rates by re-integration into the germline, resulting in heritable changes to the genome, which can occur in other species [128]. If so, this could provide a mechanism by which OA exposure increases phenotypic diversity across generations. Other processes that are more active in OA-reared crab OA triggered a heavy investment in transcriptional regulation at multiple levels of transcriptional control. DNA methylation and histone demethylase activity was upregulated in OA-reared crab, which are epigenetic mechanisms that control transcription through changes to chromatin structure and DNA accessibility [129]. A large percentage of upregulated genes (18%) were associated with the regulation of transcription from RNA polymerase II promoters. Increased expression was also detected in genes that code for post-translational gene control, such as Exportin-5, which is involved in mRNA silencing by microRNAs [130]. Increased transcription regulation activity in OA-reared crab explains the widespread changes in gene expression (44% of all genes correlated with pCO2). Given that OA resulted in widespread downregulation of a variety of biological functions (Fig 7, S4 & S5 Tables), one purpose of the transcriptional regulatory response of OA-reared crab may be to shut down (or dampen) less critical processes. Additionally, since physiological metrics were unaffected by OA [55], changes in transcription regulation may reflect physiological reprogramming that optimizes gene activity for the OA-environment, for instance by shifting metabolic pathways to improve acid/base regulation, and decreasing activity of less-critical processes to conserve energy. In addition to the broad transcriptional effects, these regulatory elements may be responsible for the unleashing of transposable element activity in response to OA [115]. Cell signaling and nervous system development was substantially more active in OA-reared crab. Many upregulated genes were associated with signal transduction, and interacted with or spanned cell membranes (e. g. Teneurin-m, Semaphorin-1A, Ankyrin-2). Increased expression of these genes suggest that OA increases the need to transfer information between the external and internal environments, particularly across the plasma membrane, to regulate activity inside the cell.

Output: contrast


Input: Claim: Curcumin, found in turmeric, has anti-inflammatory properties. [SEP] Text: As well, its anticancer activity toward mitochondria-targeted antioxidants (GSH) is closely associated with inhibition of ROS-sensitive AKT/ERK survival signaling pathways and glycolysis in human oral cancer cells. GSH is our body’s antioxidant powerhouse that is protecting our cells from free radical damage. It has been well reported that antioxidants have beneficial health effects at physiological concentrations 46. However, in normal situations, the basal level of ROS is crucially good to cell viability. It may prevent oxidative stress by maintaining the balance in ROS homeostasis, especially in cancer cells known for having much faster metabolism and higher ROS levels compared to our controls 46. Inhibition of mitochondrial superoxide may represent a novel, specific anticancer treatment strategy reducing side effects that are cytotoxic. Indeed, our work as well shows that PAC reduced mitochondrial ROS and decreased MMP. Oxidative stress is also known to induce disruption of the latter47 MMP is occurring to play a key role in mitochondrial homeostasis through selective elimination of those that are dysfunctional48. But MMP was considered as the central bioenergetic parameter for transporting ions and proteins that are indispensable for the control of normal mitochondrial function and are correlating with cell differentiation, tumorigenesis and being malignant 49. Many other studies reported that carcinoma-derived cells possess a higher mitochondrial membrane potential than normal ones 50;51 In addition, Schieke et al. (2008)52 published that higher MMP is linked with a tumorigenic property52. After inhibition of MMP by rapamycin treatment on stem cells having a high protonmotive force, its tumorigenicity decreased drastically. Moreover, our findings indicate that PAC induces intracellular ROS, generates mitochondrial superoxide and diminishes MMP leading to the release of cytochrome C in mitochondria and to the enhancement of oxidative stress in oral cancer cells. present study examined the antioral cancer and preferential killing effects of PAC, new analog of curcumine. Our results demonstrate the PAC preferential killing against oral cancer cells without effects on normal oral cells involving the regulation of proliferation, apoptosis, autophagy and oxidative stress. Therefore, PAC inhibits intracellular ROS, increased MitoSOX, diminished MMP, and PAC plays a crucial role in producing GSH, an antioxidant.

Output: pass


Input: Claim: The majority of research in graphene focuses on its application in supercapacitors and energy storage due to its high electrical conductivity. [SEP] Text: -based supercapacitors, Science 332 (2011) 1537–1542. [74] B. G. Choi, M. 3D Macroporous graphene frameworks for supercapacitors with high energy and power densities, ACS Nano 6 (2012) 4020–4028. [75] Y. Cai, A. Zhang, Y. Ping Feng, C. Zhang, Switching and rectification of a single light-sensitive diarylethene molecule sandwiched between graphene nanoribbons, J. Chem. Phys. 135 (2011) 1-6. [76] M. P. Down, C. E. Banks, Freestanding Three-dimensional graphene macroporous supercapacitor, ACS Appl. Energy Mater. 1 (2018) 891–899. [77] S. Yang, Y. Liu, Y. Hao, X. . Cao, Oxygen-vacancy abundant ultrafine Co3O4/graphene composites for high-rate supercapacitor electrodes, Adv. Sci. 5 (2018) 1-10. [78] X. Cao, Y. Shi, W. Shi, G. Lu, X. Huang, Q. Yan, Preparation of novel 3d graphene networks for supercapacitor applications, Small 7 (2011) 3163–3168. [79] C. Jiang, B. Zhao, J. Cheng, J. Li, H. Zhang, Z. Tang, J. Yang, Hydrothermal synthesis of Ni(OH)2 nanoflakes on 3D graphene foam for high-performance supercapacitors, Electrochim. Acta 173 (2015) 399–407. [80] J. Gao, H. Xuan, Y. Xu, T. Liang, X. Han, J. Yang, P. Han, D. Wang, Y. Du, Interconnected network of zinc-cobalt layered double hydroxide stick onto rGO/nickel foam for high performance asymmetric supercapacitors, Electrochim. Acta 286 (2018) 92–102. [81] X. Dong, J. Wang, J. Wang, M. B. Chan-Park, X. Li, L. Wang, W.

Output: corroborate