Over‐activation of microglia cells in the brain contributes to neurodegenerative processes promoted by the production of various neurotoxic factors including pro‐inflammatory cytokines and nitric ...oxide. Recently, accumulating evidence has suggested that mitochondrial dynamics are an important constituent of cellular quality control and function. However, the role of mitochondrial dynamics in microglial activation is still largely unknown. In this study, we determined whether mitochondrial dynamics are associated with the production of pro‐inflammatory mediators in lipopolysaccharide (LPS)‐stimulated immortalization of murine microglial cells (BV‐2) by a v‐raf/v‐myc carrying retrovirus (J2). Excessive mitochondrial fission was observed in lentivirus‐transfected BV‐2 cells stably expressing DsRed2‐mito following LPS stimulation. Furthermore, mitochondrial localization of dynamin‐related protein 1 (Drp1) (a key regulator of mitochondrial fission) was increased and accompanied by de‐phosphorylation of Ser637 in Drp1. Interestingly, inhibition of LPS‐induced mitochondrial fission and reactive oxygen species (ROS) generation by Mdivi‐1 and Drp1 knock‐down attenuated the production of pro‐inflammatory mediators via reduced nuclear factor kappa‐light‐chain‐enhancer of activated B cells (NF‐κB) and mitogen‐activated protein kinase (MAPK) signaling. Our results demonstrated for the first time that mitochondrial fission regulates mitochondrial ROS production in activated microglial cells and influences the expression of pro‐inflammatory mediators through the activation of NF‐κB and MAPK. We therefore suggest that mitochondrial dynamics may be essential for understanding pro‐inflammatory mediator expression in activated microglial cells. This could represent a new therapeutic approach for preventing neurodegenerative diseases.
LPS induced excessive mitochondrial fission through mitochondrial localization of de‐phosphorylation of Ser637 Drp1. Interestingly, inhibition of LPS‐induced mitochondrial fission and mitochondrial ROS generation by Mdivi‐1 and Drp1 shRNA attenuate the production of pro‐inflammatory mediators via reduced NF‐κB and MAPK signaling. Our results suggest that mitochondrial dynamics may be essential for understanding pro‐inflammatory mediator expression in activated microglial cells.
LPS induced excessive mitochondrial fission through mitochondrial localization of de‐phosphorylation of Ser637 Drp1. Interestingly, inhibition of LPS‐induced mitochondrial fission and mitochondrial ROS generation by Mdivi‐1 and Drp1 shRNA attenuate the production of pro‐inflammatory mediators via reduced NF‐κB and MAPK signaling. Our results suggest that mitochondrial dynamics may be essential for understanding pro‐inflammatory mediator expression in activated microglial cells.
Under endoplasmic reticulum (ER)‐stress conditions, the unfolded protein response (UPR) generates a defense mechanism in mammalian cells. The regulation of UPR signaling is important in oocyte ...maturation, embryo development, and female reproduction of pigs. Recent studies have shown that melatonin plays an important role as an antioxidant to improve pig oocyte maturation. However, there is no report on the role of melatonin in the regulation of UPR signaling and ER‐stress during in vitro maturation (IVM) of porcine oocytes. Therefore, the objective of this study was to investigate the antioxidative effects of melatonin on porcine oocyte maturation through the regulation of ER‐stress and UPR signaling. We investigated the changes in the mRNA/protein expression levels of three UPR signal genes (Bip/Grp78, ATF4, P90/50ATF6, sXbp1, and CHOP) on oocytes, cumulus cells, and cumulus‐oocyte complexes (COCs) during IVM (metaphase I; 22 hours and metaphase II; 44 hours) by Western blot and reverse transcription‐polymerase chain reaction analysis. Treatment with the ER‐stress inducer, tunicamycin (Tm), significantly increased expression of UPR markers. Additionally, cumulus cell expansion and meiotic maturation of oocytes were reduced in COCs of Tm‐treated groups (1, 5, and 10 μg/mL). We confirmed the reducing effects of melatonin (0.1 μmol/L) on ER‐stress after pretreatment with Tm (5 μg/mL; 22 hours) in maturing COCs. Addition of melatonin (0.1 μmol/L) to Tm‐pretreated COCs recovered meiotic maturation rates and expression of most UPR markers. In conclusion, we confirmed a role for melatonin in the modulation of UPR signal pathways and reducing ER‐stress during IVM of porcine oocytes.
Obesity involves chronic low‐grade inflammation within adipose tissue. Apocynin (APO) is a therapeutic agent for the treatment of inflammatory diseases. Therefore, the present study aimed to ...investigate whether APO can reduce weight gain and obesity‐induced adipose tissue inflammation. C57BL/6 mice were administered APO or orlistat (Orli) as a positive control with a high‐fat diet (HFD) for 12 weeks. Lipopolysaccharide‐stimulated 3T3‐L1 adipocytes were used for the in vitro study. Our results showed a significantly lower white adipose tissue (WAT) mass index in 10 mg/kg APO‐treated mice than in 20 mg/kg Orli‐treated mice. Moreover, the protein expression of adipose triglyceride lipase, fatty acid synthase, sterol regulatory element‐binding transcription factor 1, and peroxisome proliferator‐activated receptor γ was reversed in the WAT of 10 mg/kg APO‐treated mice. Furthermore, APO reduced the expression of the macrophage marker F4/80, decreased the mRNA levels of tumor necrosis factor‐α and monocyte chemoattractant protein‐1, and increased the mRNA levels of interleukin‐10 in WAT. APO decreased the phosphorylation of c‐Jun N‐terminal kinase, extracellular signal‐regulated kinase, and p65 in vivo and in vitro. Notably, APO had a stronger effect on the amelioration of adipose tissue inflammation than Orli did. Our findings lay the foundation for research on the use of APO as an agent to ameliorate weight gain and obesity‐induced inflammatory diseases.
We report that metal ions (M: Sn4+ and Ti4+) and boron-codoped hematite photoanodes with an n–n+ homojunction showed significantly increased photoelectrochemical (PEC) water splitting activity with ...greatly reduced surface recombination. The secondary B-doping of broadly used M-doped hematite photoanodes not only suppresses the number of M+ ions, which inevitably cause electron–hole pair (EHP) recombination, but also generates an internal electric field for easy hole extraction. Taking advantage of these effects, the maximum length (500–600 nm) of hematite, which has the reported highest PEC performance, was increased to up to 900 nm in M:B-Fe2O, which in turn increased the active area of the photoanode. The M:B-Fe2O3 with a film thickness of 900 nm and a diameter of 122 nm converted the incident photons to EHPs with substantially reduced recombination and exhibited a photocurrent density of 1.92 mA/cm2 at 1.23 VRHE. After loading inexpensive oxygen evolution reaction catalysts (FeOOH) on the surface of M:B-Fe2O3, the photocurrent density of FeOOH/M:B-Fe2O3 reached 2.35 mA/cm2 at 1.23 VRHE. The cost-effective strategy of B-doping into M-doped hematite provides a straightforward way to address the M-doping-related negative effects, such as a high electron–hole recombination rate on the surface of hematite, and thus the critical length limitation of an ideal hematite photoanode, to potentially improve the performance of PEC devices.
The proliferation of computerized functions aimed at enhancing drivers' safety and convenience has increased the number of vehicular attack surfaces accordingly. The fundamental vulnerability is ...caused by the fact that the controller area network protocol, a de facto standard for in-vehicle networks, does not support message origin authentication. Several methods to resolve this problem have been suggested. However, most of them require modification of the CAN protocol and have their own vulnerabilities. In this paper, we focus on securing in-vehicle CAN networks, proposing a novel automotive intrusion detection system (so-called VoltageIDS). The system leverages the inimitable characteristics of an electrical CAN signal as a fingerprint of the electronic control units. The noteworthy contributions are that VoltageIDS does not require any modification of the current system and has been validated on actual vehicles while driving on the road. VoltageIDS is also the first automotive intrusion detection system capable of distinguishing between errors and the bus-off attack. Our experimental results on a CAN bus prototype and on real vehicles show that VoltageIDS detects intrusions in the in-vehicle CAN network. Moreover, we evaluate VoltageIDS while a vehicle is moving.
Fucoidan has attracted attention as a potential drug because of its biological activities, which include osteogenesis. However, the molecular mechanisms involved in the osteogenic activity of ...fucoidan in human alveolar bone marrow-derived mesenchymal stem cells (hABM-MSCs) remain largely unknown. We investigated the action of fucoidan on osteoblast differentiation in hABM-MSCs and its impact on signaling pathways. Its effect on proliferation was determined using the crystal violet staining assay. Osteoblast differentiation was evaluated based on alkaline phosphatase (ALP) activity and the mRNA expression of multiple osteoblast markers. Calcium accumulation was determined by Alizarin red S staining. We found that fucoidan induced hABM-MSC proliferation. It also significantly increased ALP activity, calcium accumulation and the expression of osteoblast-specific genes, such as ALP, runt-related transcription factor 2, type I collagen-α 1 and osteocalcin. Moreover, fucoidan induced the expression of bone morphogenetic protein 2 (BMP2) and stimulated the activation of extracellular signal-related kinase (ERK), c-Jun N-terminal kinase (JNK) and p38 mitogen-activated protein kinase by increasing phosphorylation. However, the effect of fucoidan on osteogenic differentiation was inhibited by specific inhibitors of ERK (PD98059) and JNK (SP600125) but not p38 (SB203580). Fucoidan enhanced BMP2 expression and Smad 1/5/8, ERK and JNK phosphorylation. Moreover, the effect of fucoidan on osteoblast differentiation was diminished by BMP2 knockdown. These results indicate that fucoidan induces osteoblast differentiation through BMP2-Smad 1/5/8 signaling by activating ERK and JNK, elucidating the molecular basis of the osteogenic effects of fucoidan in hABM-MSCs.
Schisandra chinensis, an ancient member of the most basal angiosperm lineage which is known as the ANITA, is a fruit-bearing vine with the pharmacological effects of a multidrug system, such as ...antioxidant, anti-inflammatory, cardioprotective, neuroprotective, anti-osteoporosis effects. Its major bioactive compound is represented by lignans such as schisandrin. Molecular characterization of lignan biosynthesis in S. chinensis is of great importance for improving the production of this class of active compound. However, the biosynthetic mechanism of schisandrin remains largely unknown.
To understand the potential key catalytic steps and their regulation of schisandrin biosynthesis, we generated genome-wide transcriptome data from three different tissues of S. chinensis cultivar Cheongsoon, including leaf, root, and fruit, via long- and short-read sequencing technologies. A total of 132,856 assembled transcripts were generated with an average length of 1.9 kb and high assembly completeness. Overall, our data presented effective, accurate gene annotation in the prediction of functional pathways. In particular, the annotation revealed the abundance of transcripts related to phenylpropanoid biosynthesis. Remarkably, transcriptome profiling during fruit development of S. chinensis cultivar Cheongsoon revealed that the phenylpropanoid biosynthetic pathway, specific to coniferyl alcohol biosynthesis, showed a tendency to be upregulated at the postfruit development stage. Further the analysis also revealed that the pathway forms a transcriptional network with fruit ripening-related genes, especially the ABA signaling-related pathway. Finally, candidate unigenes homologous to isoeugenol synthase 1 (IGS1) and dirigent-like protein (DIR), which are subsequently activated by phenylpropanoid biosynthesis and thus catalyze key upstream steps in schisandrin biosynthesis, were identified. Their expression was increased at the postfruit development stage, suggesting that they may be involved in the regulation of schisandrin biosynthesis in S. chinensis.
Our results provide new insights into the production and accumulation of schisandrin in S. chinensis berries and will be utilized as a valuable transcriptomic resource for improving the schisandrin content.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Rheumatoid arthritis is an autoimmune disease that causes serious functional loss in patients. Early and accurate diagnosis of rheumatoid arthritis may attenuate its severity. Despite a diagnosis ...guideline in the 2010 American College of Rheumatology (ACR)/European League Against Rheumatism (EULAR) classification criteria for rheumatoid arthritis, the practical difficulties in its diagnosis highlight the need of developing new methods for diagnosing rheumatoid arthritis. The current study aimed to identify rheumatoid arthritis diagnostic biomarkers by using a proteomics approach. Serum protein profiling was conducted using mass spectrometry, and five distinguishable biomarkers were identified therefrom. In the validation study, the five biomarkers were quantitatively verified by multiple reaction monitoring (MRM) analysis. Two proteins, namely serum amyloid A4 and vitamin D binding protein, showed high performance in distinguishing patients with rheumatoid arthritis from healthy controls. Logistic analysis was conducted to evaluate how accurately the two biomarkers distinguish patients with rheumatoid arthritis from healthy controls. The classification accuracy was 86.0% and 81.4% in patients with rheumatoid arthritis and in healthy controls, respectively. Serum amyloid A4 and vitamin D binding protein could be potential biomarkers related to the inflammatory response and joint destruction that accompany rheumatoid arthritis.
Acute kidney injury (AKI) is a frequent complication of liver transplantation and is associated with increased mortality. We identified the incidence and modifiable risk factors for AKI after ...living-donor liver transplantation (LDLT) and constructed risk scoring models for AKI prediction. We retrospectively reviewed 538 cases of LDLT. Multivariate logistic regression analysis was used to evaluate risk factors for the prediction of AKI as defined by the RIFLE criteria (RIFLE = risk, injury, failure, loss, end stage). Three risk scoring models were developed in the retrospective cohort by including all variables that were significant in univariate analysis, or variables that were significant in multivariate analysis by backward or forward stepwise variable selection. The risk models were validated by way of cross-validation. The incidence of AKI was 27.3% (147/538) and 6.3% (34/538) required postoperative renal replacement therapy. Independent risk factors for AKI by multivariate analysis of forward stepwise variable selection included: body-mass index >27.5 kg/m2 odds ratio (OR) 2.46, 95% confidence interval (CI) 1.32-4.55, serum albumin <3.5 mg/dl (OR 1.76, 95%CI 1.05-2.94), MELD (model for end-stage liver disease) score >20 (OR 2.01, 95%CI 1.17-3.44), operation time >600 min (OR 1.81, 95%CI 1.07-3.06), warm ischemic time >40 min (OR 2.61, 95%CI 1.55-4.38), postreperfusion syndrome (OR 2.96, 95%CI 1.55-4.38), mean blood glucose during the day of surgery >150 mg/dl (OR 1.66, 95%CI 1.01-2.70), cryoprecipitate > 6 units (OR 4.96, 95%CI 2.84-8.64), blood loss/body weight >60 ml/kg (OR 4.05, 95%CI 2.28-7.21), and calcineurin inhibitor use without combined mycophenolate mofetil (OR 1.87, 95%CI 1.14-3.06). Our risk models performed better than did a previously reported score by Utsumi et al. in our study cohort. Doses of calcineurin inhibitor should be reduced by combined use of mycophenolate mofetil to decrease postoperative AKI. Prospective randomized trials are required to address whether artificial modification of hypoalbuminemia, hyperglycemia and postreperfusion syndrome would decrease postoperative AKI in LDLT.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
As the functions of vehicles are more computerized for the safety and convenience of drivers, attack surfaces of vehicle are accordingly increasing. Many attack results have shown that an attacker ...could intentionally control vehicles. Most of them exploit the vulnerability that controller area network (CAN) protocol, a de-facto standard for the in-vehicle networks, does not support message origin authentication. Although a number of methods to resolve this security vulnerability have been suggested, they have their each limitation to be applied into the current system. They have required either the modification of the CAN standard or dramatical communication load increase, which is infeasible in practice. In this paper, we propose a novel identification method, which works in the physical layer of the in-vehicle CAN network. Our method identifies electronic control units (ECUs) using inimitable characteristics of electrical CAN signals enabling detection of a malicious ECU. Unlike previous attempts to address the security problem in the in-vehicle CAN network, our method works by simply adding a monitoring unit to the existing network, making it deployable in current systems and compliant with required CAN standards. Our experimental results show that our method is able to correctly identify ECUs. In case of misclassfication rate for ECU idnetification, our method yields 0.36% in average which is approximate four times lower than the method proposed by P.-S. Murvay et al. This paper is also the first to identify potential attack models that systems should be able to detect.
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