ALBERT

Beschreibung: 〈p〉Publication date: 1 October 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 517, Issue 4〈/p〉 〈p〉Author(s): Yi Jiang, Ya-Ping Feng, Lun-Xian Tang, Yan-Li Yan, Jian-Wen Bai〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Inflammatory responses play a critical role in left ventricular remodeling after acute myocardial infarction (AMI). NR4A3, a member of the NR4A orphan nucleus receptor family, has recently emerged as a therapeutic target for treatment of inflammation. This aim of this study is to explore the therapeutic effect of NR4A3 in cardiac remodeling post AMI. Male C57BL/6 mice were administered with lentiviral over-expression of NR4A3 (lenti-NR4A3) or empty vector (lenti-con) 7 days before coronary artery ligation. H9c2 cardiomyocytes deprived of serum were used to mimic ischemic conditions i〈em〉n vivo.〈/em〉 Lenti-NR4A3 treatment significantly repressed neutrophil infiltration in the myocardium, reduced infarct size, and attenuated the reduction of left ventricular function after AMI. Furthermore, NR4A3 over-expression inhibited the NF-κB (IκB) signaling by decreasing IκBα phosphorylation and by inhibiting the translocation of p65 to the nucleus. Meanwhile, NR4A3 over-expression also increases the activity of JAK2-STAT3 signaling in mouse hearts after AMI. The inhibitory effect of NR4A3 on NF-κB activation was almost completely abolished by the JAK2 inhibitor AG490, indicating that NR4A3 prevented serum deprivation induced NF-κB activation in a STAT3 dependent manner. These findings provide novel evidence that NR4A3 could inhibit post-AMI inflammation responses via JAK2-STAT3/NF-κB signaling and may well be a therapeutic target for cardiac remodeling after AMI.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 1 October 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 517, Issue 4〈/p〉 〈p〉Author(s): Jin Young Lee, So Jeong Park, Sun Ae Han, Seung Hun Lee, Jung-Min Koh, Mark W. Hamrick, Beom-Jun Kim〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Recently, muscle has received much attention as an endocrine organ regulating other biological targets, including the pancreas, liver, and adipose tissue. Although there is a possibility that muscle-secreting factors biochemically affect bone metabolism in a paracrine manner, the net effects of myokines on the biology of osteoclasts and osteoblasts, particularly on bone mass 〈em〉in vivo〈/em〉, have not yet been thoroughly investigated. Therefore, we performed 〈em〉in vitro〈/em〉 as well as animal experiments using conditioned media (CM) collected from C2C12 myoblast and myotube cultures to better understand the interactions between muscle and bone. Compared with non-CM (i.e., control) and myoblast CM, myotube CM markedly inhibited 〈em〉in vitro〈/em〉 bone resorption through the suppression of osteoclast differentiation and resorptive activity of individual osteoclasts. Consistently, the expressions of osteoclast differentiation markers, such as tartrate-resistant acid phosphatase (〈em〉Trap〈/em〉) and calcitonin receptor (〈em〉Ctr〈/em〉), decreased with myotube CM. Myotube CM significantly stimulated preosteoblast viability and migration and reduced apoptosis, thereby resulting in an increase in calvaria bone formation. Importantly, systemic treatment with myotube CM for 4 weeks increased bone per tissue volume by 30.7% and 19.6% compared with control and myoblast CM, respectively. These results support the hypothesis that muscle plays beneficial roles in bone health via secretion of anabolic factors, in addition to mechanical stimuli, and importantly indicate that muscle-derived factors can be potential therapeutic targets against metabolic bone diseases.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 1 October 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 517, Issue 4〈/p〉 〈p〉Author(s): Alina Botvynko, Alena Bednářová, Svatopluk Henke, Nikolay Shakhno, Ladislav Čurda〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Galactooligosaccharides (GOS) are currently attracting considerable interest as prebiotic substances and can be prepared by transgalactosylation reactions from lactose using β-galactosidase. We applied various combinations of the commercial β-galactosidases, such as Nola Fit 5500, Saphera 2600 L, Maxilact LGI 5000 and Maxilact A4 MG to achieve the highest yield of GOS and reduced lactose content. The combination of the Maxilact LGI 5000 and Nola Fit 5500 resulted in amount of GOS 105 g L〈sup〉−1〈/sup〉 with lactose content lower than 5 g L〈sup〉−1〈/sup〉, whilst the combination of the Maxilact A4 MG and Maxilact LGI 5000 enzymes led to an increase in GOS to 141,1 g L〈sup〉−1〈/sup〉 and decrease of the lactose content to 46,9 g L〈sup〉−1〈/sup〉. The combination of enzymes produced a higher yield of GOS, reduced the concentration of lactose, eventually, increases the efficiency of galactooligosaccharides purification that could be potentially used in the further investigations.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 1 October 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 517, Issue 4〈/p〉 〈p〉Author(s): Kiyohiro Takahasi, Koji Onomoto, Masataka Horiuchi, Hiroki Kato, Takashi Fujita, Mitsutoshi Yoneyama〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Upon viral infection, retinoic acid-inducible gene-I (RIG-I)-like receptors detect viral foreign RNAs and transmit anti-viral signals via direct interaction with the downstream mitochondrial adaptor molecule, interferon (IFN)-β promoter stimulator-1 (IPS-1), to inhibit viral replication. Although IPS-1 is known to form prion-like oligomers on mitochondria to activate signaling, the mechanisms that regulate oligomer formation remain unclear. Here, we identified an autoinhibitory domain (AD) at amino acids 180–349 to suppress oligomerization of IPS-1 in a resting state and regulate activation of downstream signaling. Size exclusion chromatography (SEC) analysis demonstrated that AD was required to suppress auto-oligomerization of the caspase recruitment domain (CARD) of IPS-1 via intramolecular interactions. This was supported by the observation that cleavage of a peptide bond between IPS-1 CARD and AD by Tobacco Etch virus (TEV) protease relieved autoinhibition. Conversely, deletion of this domain from IPS-1 enhanced signal activation in IFN-reporter assays, suggesting that IPS-1 AD played a critical role in the regulation of IPS-1-mediated anti-viral signal activation. These findings revealed novel molecular interactions involved in the tight regulation of innate anti-viral immunity.〈/p〉〈/div〉 〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉Hypothetical model of IPS-1 autoinhibition. In the resting state, IPS-1 CARD (blue circle) interacts with the disordered region between CARD and TM (AD: light blue rectangle) to block oligomerization-induced signal transduction. Once RIG-I releases tandem-CARDs (green circle), IPS-1 CARD initiates robust oligomerization, and multiple interacting sites for downstream signaling factors (TBM: blue arrow) are revealed.〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0006291X19314652-fx1.jpg" width="384" alt="Image 1" title="Image 1"〉〈/figure〉〈/p〉〈/div〉

Beschreibung: 〈p〉Publication date: 1 October 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 517, Issue 4〈/p〉 〈p〉Author(s): Huabo He, Wei Wu, Zhener Sun, Lixin Chai〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Gastric cancer (GC) has been recognized as the major reason for global cancer-associated mortality. SEC62 homolog, preprotein translocation factor (SEC62) has been documented to possess carcinogenic functions in cancers, but its influence on GC remains elusive. Present study aimed to uncover the impact and mechanism of SEC62 in GC. We validated the upregulation of SEC62 in GC samples by GEPIA, and revealed its high level in GC cell lines. Functionally, depletion of SEC62 hindered proliferation and encouraged apoptosis in GC cells. Furthermore, we found through Starbase 3.0 and validated that methyltransferase like 3 (METTL3) interacted with SEC62 to induce the m〈sup〉6〈/sup〉A on SEC62 mRNA, therefore facilitated the stabilizing effect of IGF2 binding protein 1 (IGF2BP1) on SEC62 mRNA. Moreover, we predicted through miRmap and validated that miR-4429 targeted and inhibited METTL3 to repress SEC62. Rescue assays demonstrated that miR-4429 inhibited GC progression through METTL3/SEC62 axis. Together, our study firstly revealed that miR-4429 prevented gastric cancer progression through targeting METTL3 to inhibit m〈sup〉6〈/sup〉A-caused stabilization of SEC62, indicating miR-4429 as a promising target for treatment improvement for GC.〈/p〉〈/div〉 〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0006291X19314123-fx1.jpg" width="230" alt="Image 1" title="Image 1"〉〈/figure〉〈/p〉〈/div〉

Beschreibung: 〈p〉Publication date: 8 October 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 518, Issue 1〈/p〉 〈p〉Author(s): Ting Lan, Ji Pang, Zhongcheng Wang, Ying Wang, Hai Qian, Yongchang Chen, Yan Wu〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Our previous study demonstrated that type II cGMP-dependent protein kinase (PKG II) inhibited epidermal growth factor (EGF) induced tyrosine phosphorylation/activation of the EGF receptor (EGFR). This paper was designed to investigate the mechanism of the inhibition of PKG II on EGFR activation. Gastric cancer cells HGC-27 and AGS were infected with an adenoviral vector encoding the cDNA of PKG II (Ad-PKG II) to overexpress PKG II and treated with 8-(4-chlorophenylthio) guanosine-3′,5′-cyclic monophosphate (8-pCPT-cGMP) to activate the kinase. Co-immunoprecipitation (Co-IP) and bimolecular fluorescence complementation (BiFC) assay were performed to detect the interaction between PKG II and EGFR. Western blotting, mass spectrometry (MS) and site mutagenesis were performed to detect the PKG II-specific phosphorylation site on EGFR. The results showed that in living COS-7 cells, which were infected with Ad-PKG II and treated with 8-pCPT-cGMP, there was an interaction between PKG II and EGFR. The results also showed that PKG II caused threonine 669 (T669) phosphorylation of EGFR in HGC-27 and AGS cells infected with Ad-PKG II and treated with 8-pCPT-cGMP, and then inhibited the activation of EGFR. When T669 of EGFR was mutated to alanine, the inhibitory effect of PKG II on the activation of EGFR was eradicated. These findings suggested a PKG II-specific phosphorylation site on EGFR, and might be beneficial to illuminate the anti-tumor role of PKG II.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 1 October 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 517, Issue 4〈/p〉 〈p〉Author(s): Jun Wang, Chang Cai, Dekang Nie, Xu Song, Guan Sun, Tongle Zhi, Bing Li, Juxing Qi, Jianyong Zhang, Honglin Chen, Qiong Shi, Rutong Yu〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Fyn-related kinase (FRK), a member of the Src-related tyrosine kinase family, functions as a tumor suppressor in several malignancies. We previously showed that FRK overexpression inhibited the growth of glioma cells. However, it is unknown whether FRK is equally effective against intracranial glioma 〈em〉in vivo〈/em〉, and the mechanism by which FRK influences glioma cell growth remains unclear. In this study, we found that tumor volume was reduced by about one-third in mice with FRK overexpression, which showed improved survival relative to controls. Immunofluorescence analysis revealed that FRK overexpression inhibited glioma cell proliferation and induced their apoptosis. Importantly, 〈em〉in vitro〈/em〉 we further found that FRK decreased the expression of integrin subunit β1 (〈em〉ITGB1〈/em〉) at both the mRNA and protein levels. FRK also inhibited transactivation by ITGB1, resulting in the suppression of its target proteins AKT and focal adhesion kinase (〈em〉FAK〈/em〉). ITGB1 overexpression promoted glioma cell growth and partially reduced FRK-induced growth suppression. These results indicate that FRK inhibits human glioma growth via regulating ITGB1/FAK signaling and provide a potential therapeutic target for the treatment of glioma.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 1 October 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 517, Issue 4〈/p〉 〈p〉Author(s): Zhuohui Song, Shufen Li, Cuiying Zhang, Li Yuan, Lingna Han, Yan Liu〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉The objective of this study was to investigate the exact therapeutic effects of Verapamil on lipopolysaccharide (LPS)-induced acute lung injury (ALI) and the molecular mechanism involved, through using LPS-induced animal models as well as LPS-stimulated mouse primary peritoneal macrophages models. Our results demonstrated that Verapamil reduced LPS-induced pathological damage of the lung tissue, infiltration of inflammatory cells and the production of IL-1β, TNF-α, and MCP-1 in the serum. The MPO activity, MDA content, lung wet/dry ratio and LDH activity were also attenuated by Verapamil. In addition, Verapamil attenuated LPS-induced inflammatory cytokine production and oxidative stress in primary murine peritoneal macrophages 〈em〉in vitro〈/em〉. Moreover, we confirmed that NF-κB/NLRP3 pathway was involved in the therapeutic effect of Verapamil against LPS-induced injury 〈em〉in vivo〈/em〉 and 〈em〉in vitro〈/em〉. In conclusion, these findings indicate that Verapamil has a therapeutic effect on LPS-induced ALI in mice. The mechanism may be related to the inhibition of NF-κB and NLRP3 signaling pathways. Verapamil may be a potential therapeutic agent for the treatment of ALI.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 1 October 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 517, Issue 4〈/p〉 〈p〉Author(s): Ifeoluwapo Matthew Joshua, Thomas Höfken〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉p21-activated kinases (PAKs) are important signalling molecules with a wide range of functions. In budding yeast, the main PAKs Ste20 and Cla4 regulate the response to hyperosmotic stress, which is an excellent model for the adaptation to changing environmental conditions. In this pathway, the only known function of Ste20 and Cla4 is the activation of a mitogen-activated protein kinase (MAPK) cascade through Ste11. This eventually leads to increased transcription of glycerol biosynthesis genes, the most important response to hyperosmotic shock. Here, we show that Ste20 and Cla4 not only stimulate transcription, they also bind to the glycerol biosynthesis enzymes Gpd1, Gpp1 and Gpp2. Protein levels of Gpd1, the enzyme that catalyzes the rate limiting step in glycerol synthesis, positively correlate with glucose availability. Using a chemical genetics approach, we find that simultaneous inactivation of 〈em〉STE20〈/em〉 and 〈em〉CLA4〈/em〉 reduces the glucose-induced increase of Gpd1 levels, whereas the deletion of either 〈em〉STE20〈/em〉 or 〈em〉CLA4〈/em〉 alone has no effect. This is also observed for the hyperosmotic stress-induced increase of Gpd1 levels. Importantly, under both conditions the deletion of 〈em〉STE1〈/em〉1 has no effect on Gpd1 induction. These observations suggest that Ste20 and Cla4 not only have a role in the transcriptional regulation of 〈em〉GPD1〈/em〉 through Ste11. They also seem to modulate 〈em〉GPD1〈/em〉 expression at another level such as translation or protein degradation.〈/p〉〈/div〉 〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0006291X19314263-fx1.jpg" width="284" alt="Image 1" title="Image 1"〉〈/figure〉〈/p〉〈/div〉

Beschreibung: 〈p〉Publication date: 1 October 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 517, Issue 4〈/p〉 〈p〉Author(s): Yujin Lee, Young Hye Kwon〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉In several cancer cells, luteolin (3′,4′,5,7-tetrahydroxyflavone) exerts anticancer effects by upregulation of oxidative stress and endoplasmic reticulum (ER) stress, which are shown to activate p53-dependent cell death. Since luteolin-mediated ER stress regulation has not been investigated in hepatocellular carcinoma (HCC) cells, we investigated the role of ER stress in anti-carcinogenic effects using p53-wild type and p53-null HCC cells treated with luteolin. Trypan blue exclusion test was implemented to determine cell viability. Western blot was applied to compare the difference of autophagy, apoptosis, and proliferation event between cell lines. ER stress and p53 activation were determined by RT-PCR. Our results showed that luteolin at 5–10 μmol/L induced higher cytotoxicity in p53-null Hep3B cells than in p53-wild type HepG2 cells. Cytotoxicity was not observed in normal liver cells. Apoptosis activation and proliferation inhibition occurred in only p53-null Hep3B cells in response to luteolin treatment. Also, luteolin induced oxidative stress and ER stress in p53-null Hep3B cells. Although we observed the induction of p21 in Hep3B cells, the concomitant increases in mRNA levels of p53 family members, including TAp63 and TAp73, were not observed. Furthermore, luteolin induced autophagy in Hep3B cells only, which enhanced cell viability. Taken together, this study suggests that luteolin-induced ER stress may exert anticancer effects in a p53-independent manner.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 1 October 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 517, Issue 4〈/p〉 〈p〉Author(s): Kan Li, Tian-Xi Liu, Jun-Feng Li, Yan-Rong Ma, Ming-Long Liu, Yi-Qing Wang, Rong Wu, Bin Li, Long-Zhen Shi, Cheng Chen〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉 〈h6〉Background〈/h6〉 〈p〉Nowadays, people diagnosed sepsis may develop acute kidney injury (AKI), resulting heavy burden of health care. Recombinant human erythroprotein (rhEPO) has been suggested to have multifunction and may be used in the prevention or treatment of AKI, and its underlying mechanism remains largely unknown.〈/p〉 〈/div〉 〈div〉 〈h6〉Methods〈/h6〉 〈p〉In our study, cell model induced by LPS-activated cell apoptosis 〈em〉in vitro〈/em〉 and AKI animal model caused by lipopolysaccharide (LPS) injection 〈em〉in vivo〈/em〉. MTT assay and Flow Cytometry were conducted to analyze cell viability and apoptosis, respectively. Western bot was used to analyze expressions of apoptosis and autophagy associated proteins, and effects on AMPK/SIRT1 pathway.〈/p〉 〈/div〉 〈div〉 〈h6〉Results〈/h6〉 〈p〉Our results suggested that rhEPO inhibited LPS-induced cell apoptosis in HK-2 and HEK-293. Moreover, we found that rhEPO activated autophagy to prevented cell apoptosis, changing the expression level of autophagy associated proteins such as LC3-I/LC3-II and P62, and AMPK/SIRT1 pathway was involved in its regulation. Additionally, both EX527 (SIRT1 inhibitor) and Compound C (AMPK inhibitor) blocked the autophagy effects caused by rhEPO and thus reversed the anti-apoptotic effects of rhEPO. Furthermore, our data demonstrated that rhEPO inhibited LPS-induced kidney tubular injury and decreased the expression level of apoptotic proteins by altering the expression level of autophagy related proteins and AMPK/SIRT1 pathway related proteins 〈em〉in vitro〈/em〉.〈/p〉 〈/div〉 〈div〉 〈h6〉Conclusion〈/h6〉 〈p〉Collectively, rhEPO suppressed LPS-induced cell apoptosis via AMPK/SIRT1 pathway mediated autophagy, and modulating their levels may serve as potential way in preventing AKI.〈/p〉 〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 24 September 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 517, Issue 3〈/p〉 〈p〉Author(s): Hongxiang Hong, Yuyu Sun, Hongjian Deng, Kun Yuan, Jinyu Chen, Wei Liu, Zhiming Cui〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Dexamethasone (DEX) induces significant cytotoxicity to human osteoblasts. cPWWP2A is recently-indentified novel circular RNA (circRNA), acting as an endogenous sponge of microRNA-579 (miR-579). The present study tested the expression and potential functions of the cPWWP2A-miR-579 axis in DEX-treated osteoblasts. We show that cPWWP2A is downregulated in the necrotic femoral head tissues of DEX-taking human patients as well as in DEX-treated human osteoblasts. In OB-6 osteoblastic cells and primary human osteoblasts ectopic overexpression of cPWWP2A potently inhibited DEX-induced miR-579 accumulation, cell death, apoptosis and programmed necrosis. Silencing miR-579, by targeted siRNAs, also attenuated DEX-induced cytotoxicity in human osteoblasts. Significantly, mimicking DEX-induced actions, cPWWP2A silencing or forced miR-579 overexpression induced significant cytotoxicity in human osteoblasts. Further analyses demonstrated that miR-579's targets, including SIRT1 and PDK1 (phosphoinositide-dependent protein kinase 1), were downregulated in DEX-treated osteoblasts. Their levels were decreased as well in the necrotic femoral head tissues of DEX-taking human patients. Taken together we show that dysregulation of the cPWWP2A-miR-579 axis is involved in DEX-induced cytotoxicity in human osteoblasts.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 24 September 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 517, Issue 3〈/p〉 〈p〉Author(s): Rui Yang, Yi Zhu, Ying Wang, Wenqi Ma, Xiqiong Han, Xin Wang, Naifeng Liu〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Osteogenic differentiation of VSMC is one of the main causes of diabetic vascular calcification, and AGEs accumulation accelerates the calcification of VSMCs in diabetic patients. Autophagy has also been found to play an important role in the process of vascular calcification. However, the potential link between AGEs, autophagy and vascular calcification is still unclear and was investigated in this study. Primary VSMCs were isolated from the thoracic aorta of Sprague Dawley rats and cultured with AGEs-BSA to induce osteogenic differentiation. VSMCs calcification was evaluated by measuring the calcium content, RUNX2 protein levels, and by Alizarin red S staining. We demonstrated that treatment of VSMCs with AGE-BSA increased the expression of HIF-1α and PDK4. AGE-BSA treatment increased LC3-II and decreased p62 protein levels. AGE-BSA exposure enhanced autophagic flux determined by mRFP-GFP-LC3 adenovirus, induced co-localization of LC3-II and LAMP-1, and increased the number of autophagasome under TEM. HIF-1α/PDK4 pathway was activated during AGEs-induced autophagy of VSMCs. In addition, autophagy played a protective role during AGE-induced calcification of VSMCs. In conclusion, AGEs enhance autophagy via the HIF-1α/PDK4 signaling pathway, and autophagy helps attenuate AGE-induced calcification of VSMCs.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 24 September 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 517, Issue 3〈/p〉 〈p〉Author(s): Sang Hwan Lee, Nayeon Lee, Subin Kim, Junghun Lee, Wooshik Choi, Seung Shin Yu, Jin Hong Kim, Sunyoung Kim〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Hepatocyte growth factor (HGF) is a versatile neurotrophic factor that mediates a variety of cellular activities. In this study, we investigated the effects of intramuscularly injected recombinant AAV vectors expressing HGF in two pathologic conditions: the sciatic nerve crush and the SOD1-G93A transgenic mouse models. AAV serotype 6 (rAAV6) was chosen based on its expression levels in, and capability of moving to, the spinal cord from the injected muscle area. In the nerve crush model, rAAV6-HGF was shown to reduce the degree of mechanical allodynia, increase the cross-sectional area of muscle fibers, promote regrowth of peripheral axons, and improve motor functions. In the SOD1-G93A TG mouse model, rAAV6-HGF increased the mass of the tibialis anterior and gastrocnemius, alleviated disease symptoms, and prolonged survival. Improvements in integrity and functions of muscle in these models seemed to have come from the ability of HGF produced from rAAV6-HGF to regulate the expression of various atrogenes through the control of the FOXO signaling pathway. Our findings suggested that intramuscular injection of rAAV6-HGF might be used to relieve various symptoms associated with muscle atrophy and/or nerve damages observed in a majority of neuromuscular diseases.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 24 September 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 517, Issue 3〈/p〉 〈p〉Author(s): Yu Zheng, Jianhao Cheng, Lei Dong, Xiaoxu Ma, Qingyao Kong〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Exposure to environmental electromagnetic fields, especially to the extremely low-frequency (ELF 〈 300 Hz) electromagnetic fields (EMFs) might produce modulation effects on neuronal activity. Long-term changes in synaptic plasticity such as long-term potentiation (LTP) involved in learning and memory may have contributions to a number of neurological diseases. However, the modulation effects of ELF-EMFs on LTP are not yet fully understood. In our present study, we aimed to evaluate the effects of exposure to ELF-EMFs on LTP in hippocampal CA1 region in rats. Hippocampal slices were exposed to magnetic fields generated by sXcELF system with different frequencies (15, 50, and 100 Hz [Hz]), intensities (0.5, 1, and 2 mT [mT]), and duration (10 s [s], 20 s, 40 s, 60 s, and 5 min), then the baseline signal recordings for 20 min and the evoked field excitatory postsynaptic potentials (fEPSPs) were recorded. We found that the LTP amplitudes decreased after magnetic field exposure, and the LTP amplitudes decreased in proportion to exposure doses and durations, suggesting ELF-EMFs may have dose and duration-dependent inhibition effects. Among multiple exposure duration and doses combinations, upon 5 min magnetic field exposure, 15 Hz/2 mT maximally inhibited LTP. Under 15 Hz/2 mT ELF-EMFs, LTP amplitude decreases in proportion to the length of exposure durations within 5 min time frame. Our findings illustrated the potential effects of ELF-EMFs on synaptic plasticity and will lead to better understanding of the influence on learning and memory.〈/p〉〈/div〉 〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0006291X19314391-fx1.jpg" width="402" alt="Image 1" title="Image 1"〉〈/figure〉〈/p〉〈/div〉

Beschreibung: 〈p〉Publication date: 24 September 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 517, Issue 3〈/p〉 〈p〉Author(s): Qing Shan, Xiaoyu Li, Mei Zheng, Xi Lin, Guotao Lu, Dongming Su, Xiang Lu〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Doxorubicin (DOX) is an antitumor drug widely used in hematological tumors and various solid tumors. However, the cardiotoxicity elicited by DOX severely limits its clinical treatment. Dimethyl itaconate (DI), a common form of itaconate, is found many potential targets for prevent heart injury. Here we employed wild type and Nrf2 knockout mice and induced a cardiotoxicity model by administration of DOX to clarify the effects of DI. After treatment with DI, we found that it could effectively alleviate the cardiotoxicity by analyzing morphology, LDH levels and heart weight/body weight ratio changes. Meanwhile we demonstrated that RIP3, a key protein of necrosis, was significantly decreased in DI treated group. Further we observed that treatment with DI could suppress oxidative stress by altering Nrf2/HO-1. Compared with vehicle group, DI could increase the tissue SOD and GSH, and reduce MDA levels, then DHE staining revealed that the level of ROS in DI group reduced by half. Finally, transmission electron microscope (TEM) data showed that treatment with DI obviously decreased the mitochondrial damage. While Nrf2 was ablated in mice, the protective effects of DI were vanished and SOD, GSH, MDA became unchanged related to vehicle group. This report provides the evidence for the protective effects of DI treatment in cardiotoxicity induced by DOX. On mechanisms, DI could reduce the oxidative stress by altering Nrf2/HO-1 pathway and prevent mitochondrial from damage. Taken together, these findings of this paper will afford the new therapeutic targets in DOX related cardiotoxicity.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 24 September 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 517, Issue 3〈/p〉 〈p〉Author(s): Chao Yuan, Pan Yang, Junyue Wang, Lingyan Jiang〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉〈em〉Citrobacter〈/em〉 species are opportunistic bacterial pathogens that are implicated in both nosocomial and community-acquired infections. Among the 〈em〉Citrobacter〈/em〉 species, 〈em〉Citrobacter koseri〈/em〉 is often isolated from clinical material, and it can cause meningitis and brain abscesses in neonates and immunocompromised individuals, thus posing a great threat to human health. However, the virulence determinants of 〈em〉C. koseri〈/em〉 remain largely unknown. Myo-inositol is an abundant carbohydrate in the environment and in certain organs of the human body, especially the brain. The 〈em〉C. koseri〈/em〉 genome harbors a cluster of genes, QCQ70420.1 to QCQ70429.1 (named the Ino-cluster in this study), which encode IolBCDE, MmsA, and an ATP-binding cassette transporter. The gene cluster may be involved in the utilization of myo-inositol. To investigate the functions of the Ino-cluster in 〈em〉C. koseri〈/em〉, we constructed a mutant strain by deleting the Ino-cluster and found that the mutant could not use myo-inositol as the sole carbon source, confirming that this cluster is responsible for myo-inositol utilization. Moreover, we investigated the function of the Ino-cluster and myo-inositol utilization in 〈em〉C. koseri〈/em〉 pathogenicity. Deletion of the Ino-cluster significantly impaired 〈em〉C. koseri〈/em〉 colonization of the brain of infected Sprague–Dawley (SD) rats and BALB/c mice, and this increased the survival rate of the infected animals, indicating that the Ino-cluster and the ability to use myo-inositol are essential for 〈em〉C. koseri〈/em〉 pathogenicity. Taken together, our findings suggest that using the Ino-cluster products, 〈em〉C. koseri〈/em〉 can exploit the abundant myo-inositol in the brain as a carbon source for growth, thus promoting colonization and virulence.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 24 September 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 517, Issue 3〈/p〉 〈p〉Author(s): Chengbin Wang, Xiaohui Liu, Ru He, Junhua Li, Rui Pan〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉The clinical management of pediatric acute myeloid leukemia (AML) is still challenging and identification of drugs that can enhance the efficacy of standard of care is a potential therapeutic strategy. We show that pamidronate, a FDA-approved drug used for bone disorders, is an attractive candidate for AML treatment. Pamidronate inhibits proliferation and induces apoptosis of AML cells regardless of cellular and genetic heterogeneity. Pamidronate displays selective 〈em〉anti〈/em〉-AML activity by preferentially inhibiting survival and colony formation of AML CD34〈sup〉+〈/sup〉 cells while normal bone marrow CD34〈sup〉+〈/sup〉 cells are largely unaffected. Importantly, pamidronate remarkably enhances the inhibitory effects of all tested AML standard of care at subtoxic concentration. Mechanism studies show that pamidronate inhibits protein prenylation via dual action on geranylgeranylation and farnesylation, and subsequently decreases Ras activity. The rescue studies using overexpression of constitutively active Ras further confirm that pamidronate augments the efficacy of AML standard of care through inhibiting Ras. Since pamidronate is already used in clinic, our preclinical findings suggest that it may be an effective addition to treatment armamentarium for AML.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 24 September 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 517, Issue 3〈/p〉 〈p〉Author(s): Yan Shui, Guo-Feng Liu, Zeng-Hong Xu, Guang-Yan Zhu〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Cyclin B is a ubiquitous regulatory molecule and has been implicated in mitosis and meiosis in oocytes. Phenomenon that differ in the length of 〈em〉cyclin B〈/em〉 3′UTR in crustacean has attracted much attention, although molecular details are poorly understood. The study of 3′UTR-interacting proteins could yield much information in translational regulation and the mRNA localization process. Previous studies on crayfish suggested that the 3′UTR (1300 bp) probably contains the potential regulatory sequence/motifs such as CPEs and K-box et al. In present study, using pull-down assay coupled with mass spectrometry approach allowing us to explore the potential proteins associated with the 3′UTR. We finally identified four candidate proteins including Hspg 2, Vtg, eef1a and Tuba1a, which annotated as significant roles involved in cell differentiation, lipid transporter activity, and meiotic cell cycle process. The preliminary results will contribute to the advance in understanding the translational activation of cyclin B in oocyte maturation regulation in crustacean.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 24 September 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 517, Issue 3〈/p〉 〈p〉Author(s): Wei Cai, Yang Zhang, Yan Liu, Hailin Liu, Zhijie Zhang, Zhen Su〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉MicroRNAs (miRNA) are reported to be a vital regulator of neuropathic pain. Even so, the molecular mechanisms of miRNA function on neuropathic pain development are known little. Our research was designed to investigate the role of miRNA in neuropathic pain development in rat modle set up by chronic sciatic nerve injury (CCI). Increasing miR-150 expression could significantly alleviate neuropathic pain in CCI rats. For farther researching the regulation mechanism of miR-150 on neuropathic pain, we screened AKT3 as a possible target of miR-150 by bioinformatic mechods and predicted a possible target of miR-150 in 3′-untranslated region (UTR) of AKT3 who serves as an oncogene. In rat model, the expression both of AKT3 mRNA and protein were significantly upregulated. The overexpressed miR-150 importantly repressed the level of AKT3 and simultaneously alleviate mechanical and thermal hyperalgesia in rat model. These suppressant impacts of miR-150 on neuropathic pain process can be reversed by the overexpression of AKT3. Considering all above results, our research declared that miR-150 can restrain neuropathic pain process though targeting AKT3 in vivo, suggesting that miR-150 could be the therapeutic target for neuropathic pain therapy by regulating AKT3.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 24 September 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 517, Issue 3〈/p〉 〈p〉Author(s): Leming Liao, Yongpei Lin, Qunhua Liu, Zhiwei Zhang, Yuanhong Hong, Jianguo Ni, Sai Yu, Ye Zhong〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Periprosthetic asepteic loosening, caused by wear debris, is one of the most severe complications, generally resulting in implant failure. Extensive osteoclast formation and activation are considered as the cause for periprosthetic osteolysis. However, few approaches have been approved to be used for preventing early-stage periprosthetic osteolysis. In this study, we investigated the preventive effects of CEP on titanium particles-induced osteolysis in a murine calvaria model. This inhibitory effect was confirmed to be realized by attenuating osteoclastogenesis in vivo. In addition, CEP markedly reduced wear particles-induced elevation of receptor activator of nuclear factor kappa B ligand (RANKL)/Osteoprotegerin (OPG) ratio in vivo. In conclusion, these data concluded that CEP demonstrated a preventive effect of CEP on titanium particles induced osteolysis, suggesting that CEP might be a novel therapeutic for periprosthesis loosening.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 24 September 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 517, Issue 3〈/p〉 〈p〉Author(s): Keisuke Maruyama, Hiroyuki Kaiya, Mikiya Miyazato, Noboru Murakami, Keiko Nakahara, Kouhei Matsuda〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Neuromedin U (NMU) plays important roles in energy homeostasis in rodents and birds. Previously, our group has isolated four cDNAs encoding precursor proteins of NMU from the goldfish brain and gut, and it was assumed that these transcripts are produced by alternative splicing. We have also demonstrated that intracerebroventricular (ICV) injection of putative goldfish NMU inhibits food intake. However, as native goldfish NMU has not yet been identified, we attempted to purify it from goldfish brain and gut extracts. To assess NMU activity in fractions at each purification step, we measured changes in the intracellular concentrations of Ca〈sup〉2+〈/sup〉 using HEK293 cells expressing goldfish NMU-R1 or -R2. We isolated a 25-amino-acid peptide (NMU-25) from the brain and gut and found that its primary structure is similar to that of mammalian NMU. Another 21-amino-acid peptide (NMU-21) was purified from the brain, but not from the gut. Furthermore, a 9-amino-acid peptide (NMU-9) identical to the C-terminus of NMU-21 and -25 was also isolated from the brain and gut. Treatment with synthetic NMU-9, -21 and -25 dose-dependently increased the intracellular Ca〈sup〉2+〈/sup〉 concentration in mammalian cells expressing goldfish NMU-R1 and -R2. We also examined the effect of ICV-administered synthetic goldfish NMUs on goldfish food intake. NMU-25 inhibited food intake to the same degree as NMU-21. However, the inhibitory effect of NMU-9 was slightly weaker than those of NMU-21 and -25. These results indicate that several molecular forms of NMU exist in the goldfish brain and gut, and that all of them play physiological roles via NMU-R1 and NMU-R2.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 24 September 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 517, Issue 3〈/p〉 〈p〉Author(s): Haibing Wang, Gaochenxi Zhang〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉β,β-Dimethylacrylshikonin (DMAS), an active ingredient of Lithospermum erythrorhizon and Arnebia euchroma, possess anti-neoplasm properties. Recently, DMAS was reported to stimulate autophagy in lung adenocarcinoma cells. However, the mechanisms by which DMAS modulates autophagy. have not yet been clearly elucidated. In this study, we found that DMAS significantly elevated intracellular free calcium accumulation. This activated the CaMKKβ-AMPK-mTOR pathway, subsequently inhibited mTOR and its substrate p70s6k and 4E-BP1, eventually leading to autophagy. In addition, we demonstrated that inhibition of autophagy by BAPTA-AM or STO-609 or compound C potently enhanced DMAS-induced lung adenocarcinoma cells apoptosis and growth inhibition. Overall, our results suggested that cytoprotective autophagy was triggered by DMAS via CaMKKβ-AMPK-mTOR signaling cascade in human lung adenocarcinoma cells, meaning that combining use of DMAS and autophagy inhibitors as a novel therapeutic option for lung adenocarcinoma will be very promising.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: Available online 18 August 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Sheng-Li Liu, Yan-Man Zhou, Da-Bin Tang, Neng Zhou, Wei-Wei Zheng, Zhong-Hua Tang, Cai-Wen Duan, Jing Chen〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Aplastic anemia (AA) is a serious blood system disease that threatens human health. At present, the main cause of this disease is believed to be immune hyperfunction. However, the specific metabolic mode involved in the occurrence of lymphocytes in AA is still unknown. In addition, whether rapamycin, a specific blocker of the mTOR signaling pathway, plays a therapeutic role by inhibiting lymphocyte metabolism remains unclear. We induced an AA mouse model through the classical immune-mediated pathway and simultaneously administered rapamycin intervention therapy. First, the AA-associated phenotypic changes and the efficacy of rapamycin in the treatment of AA were discussed. Second, the proliferation and metabolic pathway of bone marrow (BM) lymphocytes in AA and the effect of rapamycin on this process were determined. Finally, the expression levels of mTOR pathway-related proteins were analyzed. By inhibiting the mTOR signaling pathway, rapamycin could ameliorate the phenotype of the immune-mediated AA model and inhibit the proliferation of T cells by preventing cell cycle transition from G0 to G1 phase. Moreover, we found that mitochondrial oxidative phosphorylation is involved in the metabolic reprogramming of T cells in AA and that rapamycin can inhibit this process. We confirmed that mitochondrial oxidative phosphorylation is involved in the metabolic reprogramming of T cells in AA and further extended the mechanism of rapamycin in treating AA by inhibiting the mTOR signaling pathway. This viewpoint may provide a new therapeutic idea for clinical applications.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: Available online 18 August 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Miroslav N. Nenov, Anton E. Malkov, Maxim V. Konakov, Sergey G. Levin〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉It has been shown that pro-inflammatory cytokines preferentially attenuate long-term potentiation (LTP), at the same time the effect of anti-inflammatory cytokines on synaptic plasticity has not been fully studied yet. Here we studied the effect of two anti-inflammatory cytokines - interleukin-10 (IL-10) and transforming growth factor-β1 (TGF-β1) on long-term potentiation. It was found that exogenously added IL-10 as well as TGF-β1 were able to effectively facilitate LTP evoked with ether high frequency or theta burst stimulation protocols in CA1 area of hippocampus. Effectiveness of IL-10 and TGF-β1 on LTP varied depending on the concentration of used cytokine and type of tetanic stimulation protocol used for LTP induction. Overall the positive effect of studied cytokines on LTP was associated with their ability to increase basal synaptic strength at Schaffer collateral – CA1 synapse. At the same time IL-10 and TGF-β1 did not have any effect on short-term plasticity. Our results provide new evidence upon the modulatory effects that anti-inflammatory cytokines exert on synaptic plasticity further highlighting their potency as modulators of neuronal function.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: Available online 18 August 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Jun Ma, Xiao-li Chen, Qin Sun〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉〈em〉Mycobacterium tuberculosis〈/em〉 (MTB) infection could induce death of host human macrophages, promoting bacterial spread. In the current study we tested the potential role of microRNA-579 (miR-579) in the death of macrophages infected with MTB. In the primary human macrophages MTB infection induced upregulation of miR-579 but downregulation of its mRNA targets, 〈em〉SIRT1〈/em〉 and 〈em〉PDK1〈/em〉, which were accompanied by significant macrophage death and apoptosis. miR-579 inhibition, by its anti-sense sequence, restored SIRT1-PDK1 expression and significantly attenuated MTB-induced cytotoxicity and apoptosis in human macrophages. Conversely, ectopic overexpression of miR-579 further downregulated SIRT1-PDK1 expression and exacerbated MTB-induced cytotoxicity in human macrophages. Further studies showed that cPWWP2A, the miR-579's endogenous sponge circRNA, was downregulated in MTB-infected macrophages. Conversely, forced overexpression of cPWWP2A, by a recombinant adeno-associated virus construct, reversed MTB-induced miR-579 upregulation and macrophage cytotoxicity. Taken together, our results show that miR-579 upregulation mediates MTB-induced macrophage cytotoxicity. Targeting cPWWP2A-miR-579 axis could be a novel strategy to protect human macrophages from MTB infection.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: Available online 17 August 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Yusuke Suenaga, Mami Yamamoto, Tetsushi Sakuma, Manabu Sasada, Fumio Fukai, Miki Ohira, Yohko Yamaguchi, Takashi Yamamoto, Kiyohiro Ando, Toshinori Ozaki, Akira Nakagawara〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉〈em〉TAp63〈/em〉 is an isoform of 〈em〉p63〈/em〉 gene〈em〉, a p53〈/em〉 family gene that suppresses tumorigenesis via transcriptional regulation. TAp63 represses transcription of 〈em〉MYC〈/em〉 oncogene in glioblastomas; however, its role in another 〈em〉MYC〈/em〉 family gene, 〈em〉MYCN,〈/em〉 has remained elusive. In this study, we showed that TAp63 repressed transcription of the 〈em〉MYCN〈/em〉 gene in human cancer cells. Overexpression of TAp63 in HeLa cells suppressed 〈em〉MYCN〈/em〉 expression, whereas knockdown of TAp63 had the opposite effect. By binding to exon 1 of 〈em〉MYCN〈/em〉 gene, TAp63 suppressed the promoter activities of 〈em〉MYCN〈/em〉 and its 〈em〉cis〈/em〉-antisense gene, 〈em〉NCYM〈/em〉. Other p53 family members, p53 and TAp73, showed lesser ability to suppress 〈em〉MYCN/NCYM〈/em〉 promoter activities compared with that of TAp63. All-〈em〉trans〈/em〉-retinoic acid (ATRA) treatment of 〈em〉MYCN〈/em〉/〈em〉NCYM〈/em〉-amplified neuroblastoma CHP134 cells induced 〈em〉TAp63〈/em〉 and reduced 〈em〉p53〈/em〉 expressions, accompanied by downregulation of 〈em〉MYCN/NCYM〈/em〉 expressions. Meanwhile, TAp63 knockdown inhibited ATRA-induced repression of 〈em〉NCYM〈/em〉 gene expression. Blocking the p53 family binding sites by CRISPR-dCas9 system in CHP134 cells induced 〈em〉MYCN〈/em〉/〈em〉NCYM〈/em〉 expression and promoted apoptotic cell death. Expression levels of 〈em〉TAp6〈/em〉3 mRNA inversely correlated with those of 〈em〉MYCN/NCYM〈/em〉 expression in primary neuroblastomas, which was associated with a favorable prognosis. Collectively, TAp63 repressed 〈em〉MYCN〈/em〉/〈em〉NCYM〈/em〉 bidirectional transcription, contributing to the suppression of neuroblastoma growth.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: Available online 16 August 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Sangwoo Kim, Eun-Jung Kim, Ji-Bin Park, Seon-Won Kim, Kyung-Jin Kim〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Isoprenoids comprise a diverse group of natural products with a broad range of metabolic functions. Isoprenoids are synthesized from prenyl pyrophosphates by prenyltransferases that catalyze the isoprenoid chain-elongation process to different chain lengths. We hereby present the crystal structure of geranylgeranyl pyrophosphate synthase from the marine flavobacterium 〈em〉Nonlabens dokdonensis〈/em〉 DSW-6 (〈em〉Nd〈/em〉GGPPS). 〈em〉Nd〈/em〉GGPPS forms a hexamer composed of homodimeric trimer, and the monomeric structure is composed of 15 α-helices (α1–α15). In this structure, we observed the binding of one pyrophosphate molecule and two glycerol molecules that mimicked substrate binding to the enzyme. The substrate binding site of 〈em〉Nd〈/em〉GGPPS contains large hydrophobic residues such as Phe, His and Tyr, and structural and amino acids sequence analyses thereof suggest that the protein belongs to the short-chain prenyltransferase family.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: Available online 14 August 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Yulong Xia, Tatsukuni Ohno, Naoto Nishii, Arundhati Bhingare, Hidetake Tachinami, Yoshihisa Kashima, Shigenori Nagai, Hirohisa Saito, Susumu Nakae, Miyuki Azuma〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Interleukin-33 (IL-33) is a nuclear-associated cytokine of the IL-1 family. IL-33 and its receptor ST2 axis exert conflicting anti-tumor and pro-tumor effects in various tumors. In this study, we examined the role of endogenously produced IL-33 in the colon-26 tumor model, in which involvement of the IL-33:ST2 pathway was negligible on the tumor side. We found that the generation of regulatory T cells (Tregs) and CD8〈sup〉+〈/sup〉 T cells, and IFN-γ expression by both CD4〈sup〉+〈/sup〉 and CD8〈sup〉+〈/sup〉 T cells (T cell activation) were impaired in IL-33-deficient mice. Overall antitumor responses, assessed by tumor growth and IFN-γ expression by tumor-infiltrating CD8〈sup〉+〈/sup〉 T cells, were also impaired, even after Treg adjustment prior to tumor inoculation. These results indicate that endogenous IL-33 augmented CD8〈sup〉+〈/sup〉 T cell-mediated antitumor responses in this colon carcinoma model, with higher CD8〈sup〉+〈/sup〉 T cell-infiltration and overcoming pro-tumor effects by increased Tregs. Exogenous application of IL-33 into the tumors did not enhance CD8〈sup〉+〈/sup〉 T cell-mediated antitumor responses despite marked elevation of innate responses showing upregulation of proinflammatory cytokine/chemokine expression, neutrophil recruitment, and dendritic cell activation. Our results suggest a dual role for endogenous IL-33 in antitumor responses and suggest that the balance of CD8〈sup〉+〈/sup〉 T cells:Tregs in the tumor microenvironment is one of key factors for estimating the contribution of IL-33-mediated antitumor responses. Therefore, the development of IL-33-based cancer immunotherapy may require a target cell-specific approach.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: Available online 14 August 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Xuewen Wu, Wenxue Tang, Emil C. Muly, Li Zhang, Xi Lin〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Connexin26 (Cx26) and Cx30 are the predominant connexin subtypes found in the cochlea. They play an essential role in the cochlear functions. However, most studies use mice and the data on the cochlear expression profiles of the two Cxs in higher animals (e.g., humans) are scarce. Studies using the cochleae from non-human primate other than mice may provide information needed to narrow this gap. Here we studied cellular distributions of Cx26 and Cx30 in the adult monkey and guinea pig cochleae by immunofluorescent labeling and confocal microscopy observations. We detected Cx26 and Cx30 expressions in the type I, II& V fibrocytes in the spiral ligament, fibrocytes of the spiral limbus, in the supporting cells of organ of Corti, inner and outer sulcus cells, and in the basal cells of the stria vascularis. Both Cx26 and Cx30 were not detected in hair cells, in mesenchymal cells under the basilar membrane and cells lining the scala vestibule. Cells of the Reissner's membrane and spiral ganglion neurons are also negative. These findings demonstrate that cochlear expressions of Cx26 and Cx30 in the adult mouse, guinea pig and non-human primate have a common cellular pattern.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: Available online 14 August 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Chao Ma, Han Chen, Siming Zhang, Yingying Yan, Ronghua Wu, Yongjun Wang, Yan Liu, Liu Yang, Mei Liu〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Exosomes are a type of extracellular vesicles derived from cells and mediators of intercellular communication. Different cell types have their own unique exosomes for exchanging information. We previously found that SASH1, a tumor suppressor, was lowly expressed or absent in glioma tissues and glioma C6 cells, but the structure and function of the corresponding exosomes had been unclear. Hence, we aimed to investigate whether exosomes generated from normal glial cells and glioma cells form different protein patterns and whether those derived from normal glial cells affect 〈em〉SASH1〈/em〉 expression in glioma cells. We collected exosomes from astrocytes and C6 cells and identified their exosomal proteins through mass spectrometry. We also performed gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) analyses, whose results showed that both the total and unique exosomal proteins from each cell type were similar. Moreover, the KEGG analysis revealed different clusters of unique exosomal proteins in glial cells and glioma cells. In the normal glial cells, the top clusters were mainly involved in processes with RNA transcripts and proteins, whereas in glioma cells the clusters were attributed to PI3K-Akt signaling, cell adhesion, and cancer-related pathways. Western blot analysis showed that HMGB1 exists in exosomes derived from cultured astrocytes, although its expression was higher in glioma C6 cells. Furthermore, we found that exosomes extracted from astrocytes could increase 〈em〉SASH1〈/em〉 expression in C6 cells (〈em〉P〈/em〉 = 0.040), whereas those derived from HMGB1-depleted astrocytes could not (〈em〉P〈/em〉 = 0.6133). The expression levels of 〈em〉SASH1〈/em〉 decreased after the addition of extracellular recombinant HMGB1 protein, whereas that of TLR4 increased. Our study is the first to demonstrate that HMGB1 plays different roles depending on its form: as an extracellular protein, HMGB1 decreases 〈em〉SASH1〈/em〉 expression, but as an exosomal protein, HMGB1 increases 〈em〉SASH1〈/em〉 expression. Nevertheless, the mechanism, which partly depends on the TLR4 pathway, behind these opposing effects requires further study. Our novel findings on the structure-dependent roles of the cytokine HMGB1 in promoting or inhibiting cancer provide a fresh insight into the interactions of cancer cells with the microenvironment.〈/p〉〈/div〉 〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0006291X19315748-fx1.jpg" width="359" alt="Image 1" title="Image 1"〉〈/figure〉〈/p〉〈/div〉

Beschreibung: 〈p〉Publication date: 8 October 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 518, Issue 1〈/p〉 〈p〉Author(s): Qingliang Shen, Jae-Hyun Cho〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Nonstructural protein 1 (NS1) is a multifunctional virulence factor of influenza virus. The effector domain (ED) of influenza viruses is capable of binding to a variety of host factors, however, the molecular basis of the interactions remains to be investigated. The isolated NS1-ED exists in equilibrium between the monomer and homodimer. Although the structural diversity of the dimer interface has been well-characterized, limited information is available regarding the internal conformational heterogeneity of the monomeric NS1-ED. Here, we present the solution NMR structure of the NS1-ED W187R of the 1918 influenza A virus, which caused the “Spanish flu.” Structural plasticity is an essential property to understand the molecular mechanism by which NS1-ED interacts with multiple host proteins. Structural comparison with the NS1-ED from influenza A/Udorn/1972 (Ud) strain revealed a similar overall structure but a distinct conformational variation and flexibility. Our results suggest that conformational flexibility of the NS1-ED might differ depending on the influenza strain.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: Available online 12 August 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Hwan Choi, Hyeok Ju Park, Hongwon Kim, Junyeop Kim, Yong Kyu Lee, Jongpil Kim〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Transcription factors play a central role in pluripotency transcription circuitry for establishing pluripotent reprogramming. Master transcription factors Oct4, Nanog, and Sox2 are known to form the core of the pluripotency transcription network. Other transcription factors also play critical roles for further refining the core circuitry for pluripotency in induced pluripotent stem (iPS) cells. Here, we reported that Nac1 interacted with the master pluripotent factors Oct4 and Nanog co-occupies gene promoters bound by these transcriptional factors for establishing pluripotency. Moreover, this interaction coordinates gene expression with H3K4me3 in the somatic cell reprogramming. Knockdown of Nac1 suppressed somatic cell reprogramming, whereas overexpression of Nac1 resulted in enhanced efficiency of induced pluripotent cell generation. Altogether, these results reveal the genome wide role for Nac1 in the contribution to the pluripotency circuitry and the regulation of the establishing pluripotent state.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: Available online 10 August 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Samar Bhallabha Mohapatra, Narayanan Manoj〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Glycosyl hydrolases belonging to the family 4 (GH4) use a unique redox-based NAD〈sup〉+〈/sup〉-dependent reaction mechanism involving anionic intermediates and requires a divalent metal ion and reducing conditions for catalytic activity. These enzymes display wide specificity and selectivity for their substrates. However, the structural basis of substrate binding, recognition and specificity remains poorly studied. Here, we report the crystal structure of 〈em〉Thermotoga maritima〈/em〉 TmAgu4B, a GH4 α-glucuronidase, in complex with Co〈sup〉2+〈/sup〉 and citrate. Analysis of GH4 structures show that the metal ion is present in a conserved octahedral coordination with conserved side chain atoms, the ligand atoms and an invariant water molecule. The data provides the first structural evidence for a metal-activated hydroxide ion that acts as the general base to deprotonate the C3-hydroxyl group of the glycone, a rate-limiting step in the mechanism. Furthermore, the citrate binding mode in the active site is analogous to a bound glucuronide substrate and provides insights into the mode of substrate interaction with the metal ion, the active site residues and, the structural basis of substrate recognition in a GH4 α-glucuronidase.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 1 October 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 517, Issue 4〈/p〉 〈p〉Author(s): Jing Zhao, Quan-You Gao〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Spinal cord injury (SCI) leads to sensorimotor deficits and autonomic changes. Macrophages and microglia could be polarized into the classically activated pro-inflammatory M1 phenotype or the alternatively activated anti-inflammatory M2 phenotype. Transmembrane protein with unknown function 16F (TMEM16F) exhibits functional diversity and may contribute to microglial function. However, the effects of TMEM16F on the modulation of macrophage/microglial polarization are still not fully understood. In the study, TMEM16F up-regulation was detected after SCI in mice, and TMEM16F protein was found in macrophages/microglia in injured spinal cord sections. Depletion of TMEM16F improved motor function in male mice with SCI. M1-type macrophages/microglia accumulated in lower numbers in the injured spinal cord of TMEM16F-knockout (KO) mice. M2 polarization inhibited by SCI was improved in mice with TMEM16F deficiency. TMEM16F deletion also attenuated microglial/macrophage pro-inflammatory response. Furthermore, significant down-regulation of A disintegrin and metalloprotease 17 (ADAM17) was observed in TMEM16F–KO mice. Importantly, TMEM16F-promoted M1 polarization and -inhibited M1 polarization were largely associated with the suppression of ADAM17. Overall, our findings provided new insights into the regulatory mechanisms of macrophage/microglial polarization, thereby possibly facilitating the development of new therapeutic strategies for SCI through the regulation of TMEM16F/ADAM17 signaling.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 8 October 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 518, Issue 1〈/p〉 〈p〉Author(s): Wanrong Tang, Zhiguo Wang, Chenyun Zhang, Chao Wang, Zhenzhen Min, Xin Zhang, Dan Liu, Jiejie Shen, Xiaoling Xu〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Malyl-coenzyme A lyase (MCL) is a carbon–carbon bond lyase that catalyzes the reversible cleavage of coenzyme A (CoA) thioesters in multiple carbon metabolic pathways. This enzyme contains a CitE-like TIM barrel and an additional C-terminal domain that undergoes conformational changes upon substrate binding. However, the structural basis underlying these conformational changes is elusive. Here, we report the crystal structure of MCL from the thermophilic photosynthetic bacterium 〈em〉Roseiflexus castenholzii〈/em〉 (〈em〉Rfx〈/em〉MCL) in the apo- and oxalate-bound forms at resolutions of 2.50 and 2.65 Å, respectively. Molecular dynamics simulations and structural comparisons with MCLs from other species reveal the deflection of the C-terminal domain to close the adjacent active site pocket in the trimer and contribute active site residues for CoA coordination. The deflection angles of the C-terminal domain are not only related to the occupation but also the type of bound substrates in the adjacent active site pocket. Our work illustrates that a conformational switch of the C-terminal domain accompanies the substrate-binding of MCLs. The results provide a framework for further investigating the reaction mechanism and multifunctionality of MCLs in different carbon metabolic pathways.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 8 October 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 518, Issue 1〈/p〉 〈p〉Author(s): Dong Wook Lee, Inhwan Hwang〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Preprotein import into chloroplasts is mediated by the coordinated actions of translocons at the outer and inner envelopes of chloroplasts (Toc and Tic, respectively). The cleavable N-terminal transit peptide (TP) of preproteins plays an essential role in the import of preproteins into chloroplasts. The Tic40 protein, a component of the Tic complex, is believed to mediate the import of preproteins through the inner envelope. In this study, we aimed to obtain 〈em〉in vivo〈/em〉 evidence supporting the role of Tic40 in preprotein import into chloroplasts. Contrary to previous findings, the import of various preproteins with wild-type TPs showed no difference between 〈em〉tic40〈/em〉 and wild-type protoplasts of 〈em〉Arabidopsis thaliana〈/em〉. However, the import of N-terminal mutants of the RbcS protein (RbcS-nt), in which basic amino acid residues (arginine and lysine) in the central region of the TP were substituted with neutral (alanine) or acidic (glutamic acid) amino acid residues, was dependent on Tic40. In addition, in 〈em〉tic40〈/em〉 protoplasts, the inner envelope protein Tic40 tagged with HA (hemagglutinin) showed more intermediate form present in the stroma. Based on these results, we propose that protein can be imported into chloroplast by either Tic40-independent or Tic40-dependent pathways depending on the types of TP.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 1 October 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 517, Issue 4〈/p〉 〈p〉Author(s): Haiou Chen, Shigang Tang, Jinmao Liao, Meng Liu, Yihe Lin〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Human umbilical cord blood mesenchymal stem cells (hUCB-MSCs) have been used to facilitate healing in animal models of liver injury, while granulocyte colony-stimulating factor (G-CSF) has been shown to stimulate stem cell mobilization and these cells may contribute to liver repair. hUCB-MSCs were characterized by flow cytometry, and transplanted into rats with 〈span〉d〈/span〉-galactosamine (D-GalN)/lipopolysaccharides (LPS)-induced acute liver failure (ALF) together with granulocyte colony-stimulating factor (G-CSF). Liver function, oxidative stress and pro-inflammatory cytokines expressions were examined using enzyme-linked immunosorbent assay (ELISA). Hematoxylin-eosin (HE) staining was used to observe the morphological changes. Apoptosis was investigated by terminal dUTP nick end labeling (TUNEL) staining. Bromodeoxyuridine (BrdU) cell proliferation assay was analyzed by immunofluorescence and immunohistochemistry. In the results, cultured hUCB-MSCs displayed proliferation and adipogenic and osteogenic differentiation potentials. hUCB-MSCs in combination with G-CSF significantly attenuated ALF-induced liver function injury. Furthermore, hUCB-MSCs and G-CSF treatment remarkably suppressed the secretions of pro-inflammatory cytokines and MDA activation induced by ALF. In addition, inflammation, lesions and cell apoptosis in liver tissues were obviously ameliorated by application of hUCB-MSCs and G-CSF. In conclusion, hUCB-MSCs, alone or co-treatment with G-CSF could ameliorate ALF in rats by inhibiting liver function injury, production of pro-inflammatory cytokines, oxidative stress, and liver cell apoptosis.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 1 October 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 517, Issue 4〈/p〉 〈p〉Author(s): Fang-Feng Chiu, Chih-Hsiang Leng, Yi-Jiun Ding, Jen-Chang Chang, Li-Sheng Chang, Shu-Pei Lien, Hsin-Wei Chen, L. Kristopher Siu, Shih-Jen Liu〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉The pneumolysin (Ply) protein of 〈em〉Streptococcus pneumoniae〈/em〉 is composed of four domains and possesses several different but related activities. In this study, recombinant Ply and two truncated forms, Ply domain 1-3 and Ply domain 4 (rPly4), were expressed and characterized regarding their participation in apoptosis, the stimulation of cytokine production, hemolytic activity and virulence. rPly4 activated murine bone marrow-derived dendritic cells in a Toll-like receptor (TLR) 4-dependent manner. The rPly4 alone was able to produce hemolytic activity at high concertation and penetrate the lipid bilayer. We further demonstrated that domain 4 of Ply involved in the virulence of the bacteria in mouse model. In the absence of apoptotic activity, the virulence level caused by rPly4 was similar to that of full length Ply. Our data suggested that domain 4 of Ply alone with TLR4 agonist and hemolytic activity may play roles in virulence of 〈em〉Streptococcus pneumoniae.〈/em〉〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 1 October 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 517, Issue 4〈/p〉 〈p〉Author(s): Gang Zhao, Yahua Song, Lei Dong, Haitao Shi, Hong Li, Longbao Yang, Jinhai Wang〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Lemur tyrosine kinase 2 (LMTK2) was recently identified as a novel cancer-related gene in several human cancers. However, little is known of its function in hepatocellular carcinoma (HCC). Here we aim to investigate the expression pattern, biological function, and regulatory mechanism of LMTK2 in HCC. We found that LMTK2 was highly expressed in HCC tissues, and patients with high expression of LMTK2 in tumor tissues had shorter survival times. LMTK2 expression was also elevated in HCC cell lines, and LMTK2 silencing markedly repressed the proliferation and invasion of HCC cells. By contrast, LMTK2 overexpression exerted promotion effects on HCC cell proliferation and invasion. Our results demonstrate that LMTK2 silencing decreases the phosphorylation of glycogen synthase kinase-3β (GSK-3β) and the expression of an active β-catenin protein, leading to inhibition of Wnt/β-catenin signaling. Notably, GSK-3β inhibition significantly reversed the LMTK2 silencing-mediated antitumor effect on proliferation, invasion, and Wnt/β-catenin signaling in HCC cells. LMTK2 silencing retarded the tumor growth of HCC cells in an 〈em〉in vivo〈/em〉 xenograft tumor model, associated with downregulation of Wnt/β-catenin signaling. In conclusion, our findings suggest that silencing of LMTK2 suppresses the proliferation and invasion of HCC cells through the inhibition of Wnt/β-catenin signaling, via GSK-3β, highlighting the importance of LMTK2/GSK-3β/Wnt/β-catenin signaling in HCC progression.〈/p〉〈/div〉 〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0006291X19314901-fx1.jpg" width="395" alt="Image 1" title="Image 1"〉〈/figure〉〈/p〉〈/div〉

Beschreibung: 〈p〉Publication date: 10 November 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 505, Issue 4〈/p〉 〈p〉Author(s): 〈/p〉

Beschreibung: 〈p〉Publication date: 10 November 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 505, Issue 4〈/p〉 〈p〉Author(s): M.A. Baqué, V.V. Gioria, G.A. Micheloud, N.M.C. Casado, J.D. Claus, A.M. Gennaro〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Different spin labels were incorporated to the membranes of cultured insect UFL-AG-286 cells in order to characterize their physical properties by Electron Paramagnetic Resonance spectroscopy (EPR). The spectrum of the spin label 12-SASL incorporated to cell membranes was similar as those obtained in membrane model systems composed of eggPC/cholesterol. However, the spectrum of the spin label CSL, chemically related to cholesterol, was drastically different in the two systems. Interestingly, when cell cholesterol content was reduced using methyl beta cyclodextrin, an EPR spectrum similar to those of model membranes was obtained. The analysis of these experiments suggests the existence of cholesterol rich regions in UFL-AG-286 cell membranes.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 10 November 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 505, Issue 4〈/p〉 〈p〉Author(s): Beijun Wu, Chunxia Li, Huimeng Lei〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉SAP90/PSD95-associated proteins (SAPAPs) are one type of scaffold protein in the postsynaptic density (PSD). Scaffold proteins play an important role in synaptic function. Recently, many studies have shown that mutations associated with scaffold proteins cause dysfunction in neuronal circuitry and in behavior. SAPAP4, as a protein in the SAPAP family, may have an impact on synaptic functions and on behaviors. To test this hypothesis, mice with a genetic deletion of SAPAP4 were used in our study. SAPAP4−/− mice displayed decreased cocaine sensitivity behavior after an acute injection of 20 mg/kg cocaine. We also found that the spine density of medium spiny neurons (MSNs) in the nucleus accumbens (NAc) shell was reduced in SAPAP4−/− mice. Furthermore, SAPAP4−/− mice displayed altered synaptic transmission and a decreased frequency and amplitude of miniature excitatory postsynaptic currents (mEPSCs) in the NAc. Our findings demonstrate that SAPAP4 plays a critical role in cocaine-related behavior and in the synaptic function of the NAc.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 10 November 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 505, Issue 4〈/p〉 〈p〉Author(s): Shengkai Luo, Hua Huang〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉〈em〉In vivo〈/em〉 growth study indicates that 〈em〉Mycobacterium smegmatis〈/em〉 could utilize D-ribonate as sole carbon source under an unknown pathway. To clarify this pathway, we start with the statistical analysis of genome neighborhood networks(GNNs) of erythrulose kinase which has been approved to participate in several sugars’ degradation. In 〈em〉M. smegmatis〈/em〉, two novel dehydrogenases (3HCDH & ADH_short) and one unknown isomerase (AP_endonuc) are targeted and characterized, for the catabolism of D-ribonate in this organism, this acid sugar is firstly oxidized into 2-keto-D-ribonate by a dehydrogenase, and then sequentially isomerized to 3-keto D-ribonate by an AP_endonuc isomerase; afterward, through decarboxylation, this 3-keto sugar acid is degraded into D-erythrulose which enters a known pathway through erythrulose kinase. Additionally, several other acid sugars (L-ribonate, D/L-lyxonate, L-threonate and D-erythronate) have been proved to be catalyzed by same enzymes and proceed with a similar catabolic pathway.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 10 November 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 505, Issue 4〈/p〉 〈p〉Author(s): Chen Lei, Han Xueming, Duan Ruihang〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Diabetes mellitus is known as a main cause to induce osteoporosis. However, the pathogenesis of osteoporosis induced by diabetes has not been fully understood. MLN64 is highly homologous to the steroidogenic acute regulatory (STAR) protein, sharing the highly conserved START domain and exhibiting various biological activities. In the study, we attempted to explore the role of MLN64 in osteoporosis progression through the in vitro and in vivo studies. At first, the in vitro study suggested that MLN64 was over-expressed during the osteoclast differentiation induced by receptor activator of NF-kB ligand (RANKL). MLN64-knockdown markedly reduced the number of TRAP-positive multinucleated cells induced by RANKL, along with the down-regulation of specific genes related to osteoclastogenesis, including tartrate-resistant acid phosphatase (TRAP), nuclear factor of activated T cells (NFATc1), cathepsin K (CTSK), tartrate resistant (ACP5) and c-FOS and up-regulation of transcriptional receptor runt related transcription factor 2 (Runx2) and osteopontin (OPN). In contrast, over-expressing MLN64 significantly promoted the production of TRAP-positive multinucleated cells triggered by RANKL. Moreover, RANKL exposure led to remarkable increase in inflammatory factors, including tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), IL-6, and MMP-1, which was attenuated by the knockdown of MLN64. Additionally, the expression of phosphorylated (p)〈img src="https://sdfestaticassets-eu-west-1.sciencedirectassets.com/shared-assets/16/entities/sbnd"〉IKKα, 〈em〉p〈/em〉-IκBα and p-nuclear factor (NF)-κB stimulated by RANKL was clearly alleviated by MLN64 silence. However, RANKL-induced inflammation was accelerated by MLN64 over-expression. Further, the in vivo study suggested that streptozotocin (STZ)-caused reduction in body weight of mice was improved by the loss of MLN64. STZ-induced diabetic mice exhibited higher levels of blood alkaline phosphatase (ALP) and TRAP, while being down-regulated in MLN64-knockout mice. MLN64 deletion reversed STZ-induced trabecular deleterious effects and stimulated bone remodeling. What's more, MLN64 knockout inhibited the expression levels of TRAP, RANKL and RANK in femoral heads of STZ mice, accompanied with the repression of inflammatory response. In conclusion, the results in our study suggested that MLN64 played a critical role in the meditation of osteoclastic differentiation, and its suppression alleviated diabetic osteoporosis in STZ-induced mice. Thus, MLN64 could be served as an essential target for developing effective therapeutic strategy to prevent diabetic osteoporosis.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 10 November 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 505, Issue 4〈/p〉 〈p〉Author(s): Yulia Zhitnyuk, Peter Gee, Mandy S.Y. Lung, Noriko Sasakawa, Huaigeng Xu, Hirohide Saito, Akitsu Hotta〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉The delivery of mRNA is advantageous over DNA delivery as it is transient and does not carry the risk of genomic DNA integration. However, there are currently few efficient mRNA delivery options available, especially for hard-to-transfect cell types, and thus new delivery methods are needed. To this end, we have established a novel mRNA delivery system utilizing chimeric virus-like particles (VLPs). We generated a novel VLP by fusing protein G of Vesicular stomatitis virus (VSV-G) with a ribosomal protein L7Ae of 〈em〉Archeoglobus fulgidus〈/em〉. This system allowed the efficient delivery of EGFP mRNA which was independent from the presence of BoxC/D motif in the mRNA sequence. Our VSVG-L7Ae VLP system demonstrated high transduction efficacy in hard-to-transfect cell lines, such as human induced pluripotent stem cells (iPS cells) and monocytes. In summary, this platform may serve as an efficient and transient transgene delivery tool for an mRNA of interest.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 10 November 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 505, Issue 4〈/p〉 〈p〉Author(s): Tania Yareli Gutiérrez-López, Lennis Beatríz Orduña-Castillo, Magda Nohemí Hernández-Vásquez, José Vázquez-Prado, Guadalupe Reyes-Cruz〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Calcium sensing receptor (CaSR) activates the NLRP3 inflammasome with consequences on homeostatic responses. However, little is known about how this process is orchestrated. Since proteolysis of critical regulators of NLRP3 inflammasome contribute to its activation, we aimed to understand how CaSR stimulates proteolytic pathways to activate the NLRP3 inflammasome. We found that proteasome and lysosome-dependent mechanisms are activated by CaSR to promote the degradation of important regulators of NLRP inflammasome. The pathway involves Gαq/PLC/PKC and Gβγ/PI3K signaling cascades and IRAK1 ubiquitination. In addition, CaSR stimulates Hsp70 expression activating a chaperone-assisted protein degradation that dictates the fate of ASC, NLRP3 (NOD-like receptor family protein 3), IRAK1 and TRAF6 proteins, turning on the NLRP3 inflammasome. In response to CaSR signaling, these proteins are degraded through the combination of CUPS (chaperone-assisted ubiquitin proteasome pathway) and CAEMI (chaperone-assisted endosomal microautophagy) systems being integrated by autophagosomes (chaperone-assisted macroautophagy, CAMA), as indicated by LC3-II, a classical marker for autophagy, that is induced in the process. Furthermore, CaSR triggers the proteolytic cleavage of pro-IL-1β (IL-1β, 31 kDa) into mature IL-1β (IL-1β, 17 kDa), via the proteasome. Taken together, our results indicate that CaSR promotes NLRP3 inflammasome activation and proteolytic maturation of IL-1β by inducing CUPS and CAEMI, chaperone-assisted degradation pathways. Overall, these results support the inclusion of CaSR as an activator of homeostasis-altering molecular processes.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 10 November 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 505, Issue 4〈/p〉 〈p〉Author(s): Keita Sonoda, Teppei Matsui, Haruhiko Bito, Kenichi Ohki〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Astrocytes are known to contact with a great number of synapses and may integrate sensory inputs. In the ferret primary visual cortex, astrocytes respond to a visual stimulus with a delay of several seconds with respect to the surrounding neurons. However, in the mouse visual cortex, it remains unclear whether astrocytes respond to visual stimulations. In this study, using dual-color simultaneous 〈em〉in vivo〈/em〉 two-photon calcium imaging of neurons and astrocytes in the awake mouse visual cortex, we examined the visual response of astrocytes and their precise response timing relative to the surrounding neurons. Neurons reliably responded to visual stimulations, whereas astrocytes often showed neuromodulator-mediated global activities, which largely masked small visual responses. Administration of the selective α1-adrenergic receptor antagonist prazosin substantially reduced such global astrocytic activities without affecting the neuronal visual responses. In the presence of prazosin, astrocytes showed weak but consistent visual responses mostly at their somata. Cross-correlation analysis estimated that the astrocytic visual responses were delayed by approximately 5 seconds relative to the surrounding neuronal responses. In conclusion, our research demonstrated that astrocytes in the primary visual cortex of awake mice responded to visual stimuli with a delay of several seconds relative to the surrounding neurons, which may indicate the existence of a common mechanism of neuron–astrocyte communication across species.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 10 November 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 505, Issue 4〈/p〉 〈p〉Author(s): Jinxia Wang, Haizhen Li, Xiaoning Wang, Tao Shen, Shuqi Wang, Dongmei Ren〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Alisol B-23-acetate (AB23A), a tetracyclic triterpenoid isolated from the rhizome of 〈em〉Alisma orientale〈/em〉, has been reported to exert anti-proliferative activities in human colon, ovarian and gastric cancer cells. However, the anti-cancer effect of this compound on human lung cancer cells has not yet been thoroughly elucidated. In the present study, we investigated the effects of AB23A on the cell viability and apoptosis in human lung cancer A549 and NCI-H292 cells. The results indicated that AB23A inhibited the growth of A549 and NCI-H292 cells in dose- and time-dependent manner, however, there was only weak cytotoxicity on normal bronchial epithelial cells. The induction of apoptosis by AB23A was demonstrated by DAPI and annexin-V-FITC/PI staining. Further investigation revealed that AB23A decreased mitochondrial membrane potential (MMP) and up regulated reactive oxygen species (ROS) level. Meanwhile, the increased Bax/Bcl-2 ratio, activated caspase-3, caspase-9 and PARP were observed. In addition, AB23A increased the release of cytochrome 〈em〉c〈/em〉 from mitochondria and the translocation of apoptotic inducing factor (AIF) into nuclei. Taken together, these results indicated that AB23A induced apoptosis by activating the intrinsic pathway, and suggested that AB23A can be used as a potential modulating agent in lung cancer.〈/p〉〈/div〉 〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0006291X18321648-fx1.jpg" width="500" alt="Image 1" title="Image 1"〉〈/figure〉〈/p〉〈/div〉

Beschreibung: 〈p〉Publication date: 10 November 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 505, Issue 4〈/p〉 〈p〉Author(s): Shuo Qiu, Yachao Jia, Jin Tang, Xuanzhe Liu, Hai Hu, Tianyi Wu, Yimin Chai〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Enthesis is the region where a tendon attaches to a bone. It is a relatively vulnerable position, and in most cases surgical treatment is required upon rupture. The reconstructed enthesis is usually weaker compared to the original, and is prone to rupture again. Hypoxia-inducible factor-1 α (HIF-1α) is known to be involved in extensive activities in cells. It is inhibited under normoxic conditions, and undergoes two essential processes, hydroxylation and ubiquitination, the latter of which has been largely unexplored. Herein, we measured the levels of HIF-1α and hydroxy-HIF-1α in VH298-treated rat tendon-derived stem cells (TDSCs) by immunoblotting. We also detected the proliferation of TDSCs using CCK-8 assay and the mRNA levels of related genes by quantitative RT-PCR. The TDSCs were observed to be induced and the chondrogenic differentiation related genes were found to be enhanced. We also simulated in-vitro wounding in a scratch test and reconstructed the enthesis in a rat model of Achilles tendon by classical surgery followed by administration of phosphate buffer saline (PBS) injection or VH298 injection. We observed that HIF-1α and hydroxy-HIF-1α levels were increased in VH298-treated TDSCs in a dose- and time-dependent manner. Thirty micromolar VH298 could significantly increase cell proliferation, migration, and expression of collagen-1α, collagen-3α, decorin, tenomodulin, tenascin C genes, and chondrogenic differentiation-related genes, collagen-2α, SRY-box9, aggrecan. VH298-treated enthesis could tolerate more load-to-failure, had a better healing pattern, and activation of HIF signaling pathway. VH298 can thus enhance the functional activities of TDSCs, enhance their chondrogenic differentiation potential, and accelerate enthesis healing by inhibiting the ubiquitination of hydroxy-HIF-1α.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 10 November 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 505, Issue 4〈/p〉 〈p〉Author(s): Qian Meng, Jing Gao, Hongwen Zhu, Han He, Zhi Lu, Minhua Hong, Hu Zhou〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Dermal fibroblast is one of the major constitutive cells of skin and plays a central role in skin senescence. The replicative senescence of fibroblasts may cause skin aging, bad wound healing, skin diseases and even cancer. In this study, a label-free quantitative proteomic approach was employed to analyzing the serial passaged human skin fibroblast (CCD-1079Sk) cells, resulting in 3371 proteins identified. Of which, 280 proteins were significantly changed in early passage (6 passages, P6), middle passage (12 passages, P12) and late passage (21 passages, P21), with a time-dependent decrease or increase tendency. Bioinformatic analysis demonstrated that the chromosome condensin complex, including structural maintenance of chromosomes protein 2 (SMC2) and structural maintenance of chromosomes protein 4 (SMC4), were down-regulated in the serially passaged fibroblast cells. The qRT-PCR and Western Blot experiments confirmed that the expression of these two proteins were significantly down-regulated in a time-dependent manner in the subculture of human skin fibroblasts (HSFb cells). In summary, we used serially passaged human skin fibroblast cells coupled with quantitative proteomic approach to profile the protein expression pattern in the temporal progress of replicative senescence in HSFb cells and revealed that the down-regulation of the chromosome condensin complex subunits, such as SMC2 and SMC4, may play an important role in the fibroblast senescence.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 10 November 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 505, Issue 4〈/p〉 〈p〉Author(s): Hee-Jung Lee, Yeondong Cho, Hyeon Jeong Kang, Hanul Choi, Kang Rok Han, Chom Kyu Chong, Young Bong Kim〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Zika virus (ZIKV), a mosquito-borne flavivirus that has recently emerged globally, poses a major threat to public health. To control this emerging disease, accurate diagnostics are required for monitoring current ZIKV outbreaks. Owing to the high nucleotide sequence similarity and cross-reactivity of ZIKV with other members of the Flaviviridae family, discrimination from other flavivirus infections is often difficult in endemic areas. ZIKV NS1 induces major virus-specific antibodies and is therefore utilized as a serological marker for ZIKV diagnosis. To identify ZIKV specific epitopes for clinical application, 33 NS1 peptides that are 15–30 amino acid in length covering whole NS1 were synthesized and analyzed linear B-cell epitopes with 38 human serum samples (20 ZIKV-positive and 18 ZIKV-negative). As a result of screening, eight epitope regions were identified. In particular, the Z8 and Z14 peptides located in the β-ladder surface region showed higher levels of binding activity in ZIKV-positive sera without cross-reactivity to other flaviviruses. These identified sensitive and specific epitopes provide a tool for design of diagnostics and structure-based vaccine antigens for ZIKV infection.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 10 November 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 505, Issue 4〈/p〉 〈p〉Author(s): Emily J. Onufer, Shirli Tay, Lauren K. Barron, Cathleen M. Courtney, Brad W. Warner, Jun Guo〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Mammalian target of rapamycin complex 1 (mTORC1) is a major regulator of cell growth and proliferation through fuel sensing. Systemic inhibition of mTOR as well as manipulation of its downstream products prevent diet-induced obesity. The purpose of this study was to determine the consequences of intestine-targeted mTORC1 inhibition. To attenuate intestinal mTORC1 activity, 〈em〉Villin-Cre〈/em〉〈sup〉〈em〉ER〈/em〉〈/sup〉 mice were crossed with 〈em〉Raptor〈/em〉〈sup〉〈em〉flox/flox〈/em〉〈/sup〉 mice, creating an intestinal-specific Raptor null line (〈em〉i-Raptor −/−〈/em〉). Mice were fed a high fat diet (HFD) and compositional changes as well as food intake levels were assessed. Over a five-week time course, i-Raptor −/− mice consistently gained less body weight on a HFD compared to wildtype (WT) mice secondary to significantly reduced food intake. Importantly, the i-Raptor −/− mice did not appear to be malnourished, demonstrated by their preservation of lean body mass. i-Raptor −/− mice also maintained a normal metabolic profile without significant changes in triglyceride or fasting glucose levels. Further investigation revealed that GDF-15 mRNA expression was significantly enhanced in i-Raptor −/− enterocytes when refed with HFD after overnight starvation. In summary, our study establishes that loss of intestinal specific-mTORC1 is protective of the development of diet-induced obesity by reducing food intake without altering the metabolic profile.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 10 November 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 505, Issue 4〈/p〉 〈p〉Author(s): Chinmayi R. Kaundinya, Handanahal S. Savithri, K.Krishnamurthy Rao, Petety V. Balaji〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉〈em〉Bacillus subtilis〈/em〉 168 EpsM (UniProt id P71063) has been electronically annotated as putative acetyltransferase in the UniProt database. The gene 〈em〉epsM〈/em〉 was cloned and overexpressed in 〈em〉E. coli〈/em〉 with an N-terminal GST tag. The purified fusion protein was shown by absorption spectroscopy, autoradiography and reverse phase HPLC to catalyse the conversion of UDP-2,4,6-trideoxy-2-acetamido-4-amino glucose to UDP-2,4,6-trideoxy-2,4-diacetamido glucose, commonly known as N,N′-diacetylbacillosamine, using acetyl coenzyme A as the donor substrate. His146 was shown by site-directed mutagenesis to be essential for acetyltransferase activity. It is hypothesized that EpsC (NAD〈sup〉+〈/sup〉 dependent UDP GlcNAc 4,6-dehydratase), EpsN (PLP dependent aminotransferase) and EpsM, all of which are part of the 〈em〉eps〈/em〉 operon, are involved in the biosynthesis of N,N′-diacetylbacillosamine.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 10 November 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 505, Issue 4〈/p〉 〈p〉Author(s): Dahu Li, Houjiu Wu, Huating Dou, Li Guo, Wei Huang〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Obesity is associated with the changes in gut microbiota. The aim of present study was to investigate the effects of sweet orange essential oil (SOEO) microcapsules on body weight and gut microbiota in obese rats induced by high-fat diet. By analyzing the body weight, fat rate and the sequence of cloned microbial small-subunit ribosomal RNA genes (16S rDNA) in rats fecal samples, we found that SOEO microcapsules decreased the body weight and increased the relative abundance of 〈em〉Bifidobacterium〈/em〉 (genus-level) in gut microbiota. The analysis of endotoxin content proved that SOEO microcapsules protected gut barrier and decreased gut endotoxin levels by increasing the content of 〈em〉Bifidobacterium〈/em〉, then ameliorated low-grade inflammation, achieving the goal of losing weight. This might be the mechanism of SOEO microcapsules to lose body weight and provided a novel anti-obesity dietary supplement.〈/p〉〈/div〉 〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉In this study, we investigated the effects of sweet orange essential oil (SOEO) microcapsules on body weight and gut microbiota in obese rats induced by high-fat diet. We found that SOEO microcapsules reduced the body weight and increased the relative abundance of 〈em〉Bifidobacterium〈/em〉 (genus-level) in gut microbiota. The analysis of endotoxin content proved that SOEO microcapsules protected gut barrier and decreased gut endotoxin levels by increasing the content of 〈em〉Bifidobacteria〈/em〉, then ameliorated low-grade inflammation, achieving the goal of losing weight. This indicates that changing gut microbiota provides a novel way to lose weight.〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0006291X18321818-fx1.jpg" width="289" alt="Image 1" title="Image 1"〉〈/figure〉〈/p〉〈/div〉

Beschreibung: 〈p〉Publication date: 17 November 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 506, Issue 1〈/p〉 〈p〉Author(s): Wenfeng Shangguan, Xue Liang, Wen Shi, Tong Liu, Manman Wang, Guangping Li〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Circular RNAs (circRNAs) have emerged as novel molecules of interest in gene regulation as other noncoding RNAs, and participating in the process of many diseases. However, the expression and functions of circRNAs in Rapid atrial pacing (RAP) dog atrial tissue still unknown. 12 canines were randomly assigned to control and pacing group. RAP at 500 beats per minute was maintained 14 days in the pacing group. The expression characterization of circRNAs were revealed by high-throughput sequencing. We totally predicted 15,990 circRNAs in dog atrial tissues. Moreover, we found 146 differentially expressed circRNAs between control and RAP dogs. Five circRNAs were selected for subsequent RT-PCR validation, and four circRNAs confirmed with the high throughput sequencing analysis. GO analysis showed that the differentially expressed circRNAs might involve in the process of “structural constituent of cytoskeleton, ion channel activity”. We explored the circRNA-miRNA interaction network, and found extensive interaction among differentially expressed circRNAs and AF related miRNAs and mRNAs. Our work firstly identified the characterization of circRNAs in the dog atrial, and revealed the differentially expressed circRNAs in the RAP dog, this might lay a solid foundation on the function of circRNA in the mechanisms of AF.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 10 November 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 505, Issue 4〈/p〉 〈p〉Author(s): Jian-Zhong Zhang, Dan Chen, Li-Quan Lv, Zheng Xu, Yi-Ming Li, Jun-Yu Wang, Kai-Wei Han, Ming-Kun Yu, Cheng-Guang Huang, Li-Jun Hou〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉microRNAs (miRNAs) control several processes known to be involved in progression of aneurysm. Here, intracranial aneurysms (IAs) were surgically induced in Sprague–Dawley rats, and we found that miR-448-3p was downregulated and KLF5 was upregulated in IA rats. We identified 〈em〉Klf5〈/em〉 as a direct target of miR-448-3p in smooth muscle cells (SMCs). In addition, aneurysms size and the lumen area of the aneurysms were smaller 4 weeks after IA induction in the miR-448-3p–treated group. miR-448-3p treatment protected the wall thickness ratio and suppressed macrophage infiltration after IA induction. IAs caused a significant increase in KLF5 expression and were alleviated by miR-448-3p. Moreover, the anti-inflammatory effect of miR-448-3p was verified in lipopolysaccharide -stimulated RAW 264.7 macrophage cells. The expression levels of KLF5, MMP2, and MMP9 levels were elevated by LPS, and were attenuated by miR-448-3p. These data suggest that miR-448-3p plays the inhibitory role in IA progression, indicating that miR-448-3p overexpression is crucial for preventing the development of IA through downregulation of macrophage-mediated inflammation.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 10 November 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 505, Issue 4〈/p〉 〈p〉Author(s): Shannon L. Nowotarski, Robert P. Feehan, Christopher Presloid, Lisa M. Shantz〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Non-melanoma skin cancer (NMSC) is the most commonly diagnosed cancer in the United States. Ultraviolet-B (UVB) irradiation is the primary carcinogen responsible for stimulating NMSC development. Ornithine Decarboxylase (ODC), the first rate-limiting enzyme in the synthesis of polyamines, is upregulated in response to a variety of proliferation stimuli, including UVB exposure. Our previous studies have demonstrated regulation of ODC synthesis by the mammalian target of rapamycin complex 1 (mTORC1) in cells transformed by oncogenic Ras. The goal of these studies was to better understand the link between mTORC1 and ODC in nontransformed cells treated with UVB. We show that the ablation of mTORC1 activity by conditional knockout of its essential component Raptor led to decreased levels of ODC protein both before and after exposure to 10 mJ/cm〈sup〉2〈/sup〉 UVB. Moreover, ODC mRNA was destabilized in the absence of Raptor, suggesting post-transcriptional regulation. We have previously shown that the ODC transcript is stabilized by the RNA binding protein (RBP) human antigen R (HuR), and the intracellular localization of HuR responds to changes in mTORC1 activity. To expand these studies, we investigated whether HuR functions to regulate ODC mRNA stability after UVB exposure. Our results show an increased localization of HuR to the cytoplasm after UVB exposure in wild-type cells compared to Raptor knockout cells, and this is accompanied by greater association of HuR with the ODC transcript. These data suggest that the localization of HuR in response to UVB is influenced, at least in part, by mTORC1 and that HuR can bind to and stabilize ODC mRNA after UVB exposure in an mTORC1-dependent manner.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 2 December 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 506, Issue 4〈/p〉 〈p〉Author(s): Tomohiro Yoshimura, Kanae Saitoh, Luchuanyang Sun, Yao Wang, Shigeto Taniyama, Kenichi Yamaguchi, Takayuki Uchida, Tsutomu Ohkubo, Atsushi Higashitani, Takeshi Nikawa, Katsuyasu Tachibana, Katsuya Hirasaka〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Cachexia, observed in most cancer patients, is a syndrome that includes wasting of bodily energy reserves and is characterized by muscle atrophy and fat loss. We have previously demonstrated that isoflavones, such as genistein and daidzein, prevent muscle wasting in tumor-bearing mice. In this study, we examined the effect of morin, a flavonoid, on cachexia. The wet weight and myofiber size of muscles in Lewis lung carcinoma (LLC) cell-bearing mice fed a normal diet were decreased, compared with those in control mice fed a normal diet. In contrast, intake of morin prevented the reduction of muscle wet weight and myofiber size. Moreover, the tumor weight in mice fed the morin diet was lower than that in mice fed the normal diet. Both cell viability and protein synthetic ability of LLC cells were reduced by treatment with morin, but C2C12 myotubes were not affected. Binding assay using morin-conjugated magnetic beads identified ribosomal protein S10 (RPS10) as a target protein of morin. Consistent with the result of morin treatment, knockdown of RPS10 suppressed LLC cell viability. These results suggest that morin indirectly prevents muscle wasting induced by cancer cachexia by suppressing cancer growth via binding to RPS10.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 2 December 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 506, Issue 4〈/p〉 〈p〉Author(s): Hua Yu, Jingjing Liu, Yizhi Dong, Min Xu, Le Xu, Huaqin Guan, Xiaoru Xia, Liangxing Wang〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉 〈h6〉Background〈/h6〉 〈p〉Previous studies have found that dihydroartemisinin (DHA) has multiple functions such as anti-inflammatory, anti-tumor in addition to anti-malarial effects. Effect of DHA on monocrotaline-induced pulmonary hypertension in rats has been reported, while the specific mechanism is not known.〈/p〉 〈/div〉 〈div〉 〈h6〉Method〈/h6〉 〈p〉A hypoxic model was established with human pulmonary arterial endothelial cells (HPAECs) to investigate the possible mechanism of DHA. Effects of DHA on proliferation of HPAECs were evaluated by CCK-8 and EdU assay. Effects of DHA on cell oxidative stress, cell migration, angiogenesis, cell cycle and autophagy, as well as the possible underlying mechanism were also detected by using the established normoxia/hypoxia cell models.〈/p〉 〈/div〉 〈div〉 〈h6〉Results〈/h6〉 〈p〉DHA significantly inhibited hypoxia induced increase of HPAECs proliferation in a dose dependent manner, migratory ability and angiogenic ability. DHA also significantly reversed hypoxia induced oxidative stress as a reduction of ROS and NO, and an increase of SOD. Autophagosomes, LC3B protein and apoptotic proteins were significantly increased in DHA treated hypoxic HPAECs. Autophagy inhibitor 3-Methyladenine diminishes the anti-hypoxia effects of DHA on cell proliferation, migration, and autophagy and apoptosis protein expression in HPAECs.〈/p〉 〈/div〉 〈div〉 〈h6〉Conclusion〈/h6〉 〈p〉DHA effectively inhibits hypoxia induced increase of cell proliferation, migration, and oxidative stress in HPAECs, and autophagy may be the underlying mechanism of DHA.〈/p〉 〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 10 November 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 505, Issue 4〈/p〉 〈p〉Author(s): Peilong Dong, Song Gu, Yiqiu Jiang, Wangxiang Yao, Kaibin Zhang, Tianqi Tao, Yang Li, Wang Li, Jianchao Gui〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Lateral ankle sprains are one of the most common injuries in sports. Recently, arthroscopic lateral ligament reconstruction has been recently advocated, however no biomechanical studies and clinical application of this technique are available. In this biomechanical study, eighteen fresh-frozen cadaveric ankles were randomized into three groups: (1) intact anterior talofibular ligament (ATFL) and calcaneofibular ligament (CFL), (2) arthroscopic anatomic reconstruction of ATFL and CFL, and (3) all arthroscopic remnant-preserving reconstruction of ATFL and CFL. The specimens were then tested for stiffness and load to ultimate failure using a customized jig. In biomechanical test, the all arthroscopic remnant-preserving reconstruction of ATFL and CFL produced a reconstruction that could withstand loads to failure and stiffness similar to the arthroscopic anatomic reconstruction. However, both two reconstruction groups were much weaker than the intact, uninjured ATFL and CFL. Moreover, we used the technique of all arthroscopic remnant-preserving reconstruction of ATFL and CFL on 20 patients from September 2016 to September 2017. American Orthopaedic Foot and Ankle Society (AOFAS) scores and Anterior Talar Translation (ATT) were applied for statistical collection at preoperative and postoperative 12 months to evaluate clinical efficacy. The differences of the preoperative and postoperative 12 months AOFAS scores and ATT of patients were both statistical significant (〈em〉P〈/em〉 〈 0.01). We confirmed that all arthroscopic remnant-preserving reconstruction of ATFL and CFL exhibited positive effect, thus promoting the recovery of ankle function and had good short-term clinical effect.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 2 December 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 506, Issue 4〈/p〉 〈p〉Author(s): Hanjun Kim, Young-Jin Choi, Young-Sun Lee, Suk Young Park, Ji-Eun Baek, Ho-Kyoung Kim, Beom-Jun Kim, Seung Hun Lee, Jung-Min Koh〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Previously, we noted that SLIT3, slit guidance ligand 3, had an osteoprotective role with bone formation stimulation and bone resorption suppression. Additionally, we found that global 〈em〉Slit3〈/em〉 KO mice had smaller long bone. Skeletal staining showed short mineralized length in the newborn KO mice and wide hypertrophic chondrocyte area in the embryo KO mice, suggesting delayed chondrocyte maturation. The recombinant SLIT3 did not cause any change in proliferation of ATDC5 cells, but stimulated expressions of chondrocyte differentiation markers, such as COL2A1, SOX9, COL10A1, VEGF, and MMP13 in the cells. SLIT3 suppressed β-catenin activity in the cells, and activation of Wnt/β-catenin signaling by lithium chloride attenuated the SLIT3-stimulated differentiation markers. ATDC5 cells expressed only ROBO2 among their 4 isotypes, and the 〈em〉Robo2〈/em〉 knock-down with its siRNA reversed the SLIT3-stimulated differentiated markers in chondrocytes. Taken together, these indicate that SLIT3/ROBO2 promotes chondrocyte maturation via the inhibition of β-catenin signaling.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 2 December 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 506, Issue 4〈/p〉 〈p〉Author(s): John T. Gamble, Yuriyah Reed-Harris, Carrie L. Barton, Jane La Du, Robert Tanguay, Juliet A. Greenwood〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Glioblastoma (GBM) is a deadly disease due to its ability to quickly invade and destroy brain tissue. Slowing or stopping GBM cell progression is crucial to help those inflicted with the disease. Our lab created an embryo-larval zebrafish xenograft model as a tool to study human GBM progression in an observable brain environment. The zebrafish brain is a dynamic and complex environment providing an optimal setting for studying GBM cell progression. Here we demonstrate the ability of our model to quantitate GBM proliferation, dispersal, blood vessel association, microtumor formation, and individual cell invasion by evaluating the importance of an extracellular matrix protein, laminin alpha 5 (lama5), on U251MG cell progression. Lama5 has been implicated in cancer cell survival, proliferation and invasion and is a known adhesion site for GBM cells. While lama5 is highly expressed in endothelial cells in the brain, it is unknown how lama5 affects GBM behavior. Using a lama5 morpholino, we discovered that lama5 decreased U251MG dispersal by 23% and doubles the formation of blood vessel dependent microtumors. Despite lama5 being a known attachment site for GBM, lama5 expression had no effect on U251MG association with blood vessels. Analysis of individual U251MG cells revealed lama5 significantly lowered invasion as mobile U251MG cells traveled 32.5  μm less, invaded 5.0 μm/hr slower and initiated invasion 60% few times per cell.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 10 November 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 505, Issue 4〈/p〉 〈p〉Author(s): Yoon-Jin Lee, Jeong-Eun Oh, Sang-Han Lee〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Extracellular acidity in the tumor microenvironment contributes to chemoresistance of malignant tumors. The objective of this study was to determine anticancer effects of arctigenin, a novel anti-inflammatory lignan extracted from seeds of 〈em〉Arctium lappa〈/em〉, on acidity-tolerant prostate cancer PC-3AcT cells. The PC-3AcT cells manifested increased tolerance to low-pH media with enhanced percent cell viability and increased resistance to docetaxel compared to their parental PC-3 cells. Arctigenin alone or in combination with docetaxel induced potent cytotoxicity. Preferential sensitization of PC-3AcT cells to arctigenin was accompanied by increased cell fractions with sub-G〈sub〉0〈/sub〉/G〈sub〉1〈/sub〉 peak and annexin V-PE(+), increased ROS levels, decreased mitochondrial membrane potential and cellular ATP content, and inhibition of PI3K/Akt/mTOR pathway. A series of changes caused by arctigenin were efficiently reversed through reducing ROS levels by radical scavenger N-acetylcysteine, thus placing ROS upstream of arctigenin-driven cytotoxicity. Collectively, these results demonstrate that arctigenin can increase oxidative stress-mediated mitochondrial damage of acidity-tolerant PC-3AcT cells, suggesting that arctigenin might be a potential therapeutic candidate to overcome acidic-microenvironment-associated chemotherapeutic resistance in prostate cancer.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 10 November 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 505, Issue 4〈/p〉 〈p〉Author(s): Shengting Wang, Qian Li, Yufang Wang, Xiaoming Li, Rui Wang, Yuhua Kang, Xukai Xue, Rui Meng, Qi Wei, Xinghua Feng〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Recently, a large number of studies have shown that circular RNA (circRNA) is involved in the development and progression of human cancer. However, its role in triple-negative breast cancer (TNBC) remains largely unknown. In this study, a novel circRNA, circ-UBAP2 (hsa_circ_0001846), was shown to be markedly upregulated in TNBC. Moreover, high circ-UBAP2 expression was closely associated with larger tumour size (〈em〉p〈/em〉 = 0.003), advanced TNM stage (〈em〉p〈/em〉 = 0.005), lymph node metastasis (〈em〉p〈/em〉 = 0.002), and unfavourable prognosis (〈em〉p〈/em〉 = 0.0256). Functionally, lentivirus-mediated stable knockdown of circ-UBAP2 dramatically weakened the ability of TNBC cells to proliferate and migrate and induced apoptosis 〈em〉in vitro〈/em〉 and 〈em〉in vivo〈/em〉. Regarding the mechanism, we found that circ-UBAP2 was mainly observed in the cytoplasm and was capable of sponging miRNA-661 to increase the expression of the oncogene MTA1. Additionally, silencing of miRNA-661 or overexpression of MTA1 could partially rescue the attenuated malignant phenotype caused by circ-UBAP2 knockdown. Taken together, our data reveal that circ-UBAP2 plays a vital regulatory role in TNBC via the miR-661/MTA1 axis and may serve as a promising therapeutic target for TNBC patients.〈/p〉〈/div〉 〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0006291X18321685-fx1.jpg" width="301" alt="Image 1" title="Image 1"〉〈/figure〉〈/p〉〈/div〉

Beschreibung: 〈p〉Publication date: 10 November 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 505, Issue 4〈/p〉 〈p〉Author(s): Yahong Han, Zhenbiao Xu, Yanan Liu, Da Wei, Jing Zhang, Le Xue, Xiangmin Zhang, Jie Qin, Hongkuan Deng, Linxia Song〈/p〉 〈div xml:lang="en"〉 〈h5〉Absract〈/h5〉 〈div〉〈p〉Y-box binding protein (YB protein) is an ancient conserved multifunctional DNA/RNA-binding protein. A novel YB protein 〈em〉DjY2〈/em〉 gene from planarian 〈em〉Dugesia japonica〈/em〉 was cloned by RACE method and characterized. This cDNA contains 689 bp with a putative open reading frame of 197 amino acids. It has a predicted molecular mass of 22.14 kDa and an isoelectric point of 9.67. Whole-mount in situ hybridization and relative quantitative real-time PCR were used to study the spatial and temporal expression pattern of 〈em〉DjY2〈/em〉 in the process of planarian regeneration. Results showed that 〈em〉DjY2〈/em〉 was expressed in many parts of the body in intact planarian, but the expression level was low in head and pharynx. The transcripts of 〈em〉DjY2〈/em〉 was significantly increased both at the head parts and the tail parts after amputation, especially at the site of cutting. The spatial expression gradually recovered to the state of intact planarian with the time of regeneration. Our results indicated that 〈em〉DjY2〈/em〉 might participate in the process of regeneration in planarian.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 10 November 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 505, Issue 4〈/p〉 〈p〉Author(s): Saeko Kimura, Tomoki Uchida, Yuki Tokuyama, Mayumi Hosokawa, Junya Nakato, Atsushi Kurabayashi, Masaru Sato, Hideyuki Suzuki, Ryuhei Kanamoto, Kousaku Ohinata〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Rubisco, an enzyme for photosynthetic carbon dioxide fixation, is a major green leaf protein and known as the most abundant protein on the Earth. We found that Rubisco digested mimicking gastrointestinal enzymatic conditions exhibited anxiolytic-like effects after oral administration in mice. Based on a comprehensive peptide analysis of the digest using nanoLC-Orbitrap-MS and the structure-activity relationship of known anxiolytic-like peptides, we identified SYLPPLTT, SYLPPLT and YHIEPV [termed Rubisco anxiolytic-like peptide (rALP)-1, rALP-1(1–7) and rALP-2, respectively], which exhibited potent anxiolytic-like effects after oral administration. The anxiolytic-like effects of rALP-1/rALP-1(1–7) were blocked by a serotonin 5-HT〈sub〉1A〈/sub〉 receptor antagonist, whereas rALP-2-induced effects were inhibited by a δ-opioid receptor antagonist. In conclusion, novel Rubisco-derived anxiolytic-like peptides, rALP-1/rALP-1(1–7) and rALP-2, act via independent neural pathways.〈/p〉〈/div〉 〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0006291X18321363-fx1.jpg" width="359" alt="Image 1" title="Image 1"〉〈/figure〉〈/p〉〈/div〉

Beschreibung: 〈p〉Publication date: 10 November 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 505, Issue 4〈/p〉 〈p〉Author(s): Debora Levy, Thatiana Correa de Melo, Bianca Yumi Ohira, Maíra Luísa Fidelis, Jorge L.M. Ruiz, Alessandro Rodrigues, Sergio P. Bydlowski〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Oxysterols are 27-carbon oxidation products of cholesterol metabolism. Oxysterols possess several biological actions, including the promotion of cell death. Here, we examined the ability of several oxysterols to induce short-term death in cancerous (human breast cancer and mouse skin melanoma cells) and non-cancerous (human endothelial cells and lung fibroblasts) cell lines. We determined cell viability, Ki67 expression, cell cycle regulation, and apoptosis after 24-h incubations with oxysterols. We found that different oxysterols had different effects on the studied parameters. Moreover, the effects depended on cell type and oxysterol concentration. Three cytotoxic oxysterols (7-ketocholesterol, cholestane-3β-5α-6β-triol, and 5α-cholestane-3β,6β-diol) inhibited the S phase and stimulated the G0/G1 or G2/M phases. These oxysterols promoted apoptosis, determined with Annexin V and propidium iodide assays. These results showed that different oxysterols have cytotoxic effects depending on the cell line. The findings suggest a potential pharmacological utility of cytotoxic oxysterols.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 10 November 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 505, Issue 4〈/p〉 〈p〉Author(s): Keigo Kumagai, Cherilyn A. Elwell, Shuji Ando, Joanne N. Engel, Kentaro Hanada〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉〈em〉Chlamydia trachomatis〈/em〉 is an obligate intracellular bacterium that replicates within a membranous compartment, the inclusion, in host cells. Its intracellular life cycle requires host sphingolipids, which are in part acquired through the ER-Golgi localized ceramide transport protein (CERT). The Chlamydia-encoded inclusion membrane protein IncD is composed of two closely linked long hydrophobic domains with their N- and C-termini exposed to the host cytosol. IncD binds directly to the pleckstrin homology (PH) domain of CERT, likely redirecting ceramide to the inclusion. The precise regions of IncD required for this interaction have not been delineated. Using co-transfection studies together with phylogenetic studies, we demonstrate that both the IncD N- and C-terminal regions are required for binding to the CERT PH domain and define key interaction residues. Native gel electrophoresis analysis demonstrates that the transmembrane region of IncD forms SDS-resistant but dithiothreitol-sensitive homodimers, which in turn can assemble to form higher order oligomers through additional N- and C-terminal domain contacts. IncD oligomerization may facilitate high affinity binding to CERT, allowing 〈em〉C. trachomatis〈/em〉 to efficiently redirect host ceramide to the inclusion.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 10 November 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 505, Issue 4〈/p〉 〈p〉Author(s): Yukino Morikiri, Eri Matsuta, Hideki Inoue〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉The skin consists mostly of extracellular matrix (ECM) composed mainly of collagen, which provides a protective barrier from the environment. The skin continuously experiences harmful stress and damage. As aging progresses, the expression of various genes declines, and physiological functional deterioration occurs. The reduction of collagen accompanying aging impairs the barrier function of the skin and weakens protection from stressors. In the nematode 〈em〉Caenorhabditis elegans〈/em〉, ECM proteins turn over during aging. Older worms of longevity mutants exhibit increased collagen expression, whereas knockdown of collagen genes shortens lifespan. However, it is unclear whether the progression of aging can be delayed by increasing collagen production via an external stimulus. In this study, we examined the effects of collagen tripeptide (CTP), a collagen-derived compound, on lifespan and aging. Our results showed that CTP upregulated collagen genes via the p38 mitogen-activated protein kinase (MAPK)/SKN-1 pathway. Moreover, CTP extended lifespan and delayed aging through p38 MAPK/SKN-1 pathway. In addition, CTP also induced collagen expression via the p38 MAPK pathway in mammals. Our findings supported that external stimuli such as CTP could promote ECM youthfulness using a conserved signaling pathway, thereby contributing to suppression of aging.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 10 November 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 505, Issue 4〈/p〉 〈p〉Author(s): Atsushi Masuyama, Tomoya Mita, Kosuke Azuma, Yusuke Osonoi, Kenichi Nakajima, Hiromasa Goto, Yuya Nishida, Takeshi Miyatsuka, Masako Mitsumata, Hirotaka Watada〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Autophagy is considered as an evolutionarily conserved cellular catabolic process. Defective autophagy has been implicated in various human diseases, including cardiovascular diseases. Recently, we and others demonstrated that defective autophagy in vascular smooth muscle cells (SMCs) promotes the progression of atherosclerosis. In this study, we investigated the role of autophagy in SMCs on plaque instability 〈em〉in vivo〈/em〉. We generated mice with a defect 〈em〉atg7〈/em〉in which is an essential gene for autophagy, in SMCs by crossing 〈em〉Atg7〈/em〉〈sup〉〈em〉f/f〈/em〉〈/sup〉 mice with transgelin (〈em〉Tagln〈/em〉) 〈em〉Cre〈/em〉〈sup〉〈em〉+/0〈/em〉〈/sup〉 mice (〈em〉Atg7cKO〈/em〉). Then, 〈em〉Atg7cKO〈/em〉 and apolipoprotein E (〈em〉apoe〈/em〉)-deficient (〈em〉apoeKO〈/em〉) mice were crossed to generate 〈em〉Atg7cKO:apoeKO〈/em〉 mice. To generate a mouse model of plaque instability, we conducted to form a tandem stenosis in the carotid artery of 〈em〉Atg7cKO:apoeKO〈/em〉 mice and their controls (〈em〉apoeKO〈/em〉 mice) at the age of 10 weeks. At 5 weeks after surgery, the percentage of cross-sectional stenosis area in the operated common carotid artery of 〈em〉Atg7cKO:apoeKO〈/em〉 mice was significantly higher than that in 〈em〉apoeKO〈/em〉 mice. In addition, thrombus, which was not observed in 〈em〉apoeKO〈/em〉 mice, was frequently found in 〈em〉Atg7cKO:apoeKO〈/em〉 mice. Furthermore, the number of Berlin blue staining-positive areas, which indicated intraplaque hemorrhage, was significantly higher in 〈em〉Atg7cKO:apoeKO〈/em〉 mice than in control 〈em〉apoeKO〈/em〉 mice. Taken together, our data suggest that defective autophagy in SMCs enhances plaque instability and the risk of plaque rupture.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 10 November 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 505, Issue 4〈/p〉 〈p〉Author(s): Zongtai Feng, Huiting Zhou, Shurong Ma, Xinxian Guan, Lulu Chen, Jie Huang, Huan Gui, Xinxin Miao, Shenglin Yu, Jiang Huai Wang, Jian Wang〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Necrotizing enterocolitis (NEC) remains one of the leading causes of death in neonatal infants and new therapeutic strategies for NEC are urgently required. The immunomodulatory agent FTY720 has been shown to have protective effects in various inflammatory diseases. In this study, we hypothesized that treatment with FTY720 confers protection against experimental NEC. Experimental NEC was induced in five-day-old C57BL/6 neonatal mice by hyperosmolar formula feeding plus hypoxia and lipopolysaccharide (LPS) challenges. Induction of NEC resulted in substantial weight loss and high mortality compared to the control group, whereas FTY720 treatment significantly attenuated weight loss and improved survival in NEC-challenged neonatal mice. FTY720 treatment strongly ameliorated NEC-induced intestinal injury with reduced apoptosis and up-regulation of intestinal barrier proteins in the ileal tissues. Furthermore, FTY720 treatment abrogated NEC-initiated intestinal and systemic inflammation with markedly diminished inflammatory cytokines and chemokines. Moreover, FTY720 treatment suppressed NEC-activated CXCL5/CXCR2 axis with down-regulated expression of CXCL5 and CXCR2 at both mRNA and protein levels. Thus, we demonstrate that FTY720 protects neonatal mice against NEC-associated lethality by ameliorating intestinal injury and attenuating inflammation, possibly via its down-regulation of NEC-induced activation of intestinal CXCL5/CXCR2 axis.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 10 November 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 505, Issue 4〈/p〉 〈p〉Author(s): Prince Rajaiah Prabhu, Sakthi Devi Moorthy, Jayaprakasam Madhumathi, Satya Narayan Pradhan, Markus Perbandt, Christian Betzel, Perumal Kaliraj〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉The notoriety of parasitic nematode survival is directly related to chronic pathogenicity, which is evident in human lymphatic filariasis. It is a disease of poverty which causes severe disability affecting more than 120 million people worldwide. These nematodes down-regulate host immune system through a myriad of strategies that includes secretion of antioxidant and detoxification enzymes like glutathione-S-transferases (GSTs). Earlier studies have shown 〈em〉Wuchereria bancrofti〈/em〉 GST to be a potential therapeutic target. Parasite GSTs catalyse the conjugation of glutathione to xenobiotic and other endogenous electrophiles and are essential for their long-term survival in lymph tissues. Hence, the crystal structure of WbGST along with its cofactor GSH at 2.3 Å resolution was determined. Structural comparisons against host GST reveal distinct differences in the substrate binding sites. The parasite xenobiotic binding site is more substrate/solvent accessible. The structure also suggests the presence of putative non-catalytic binding sites that may permit sequestration of endogenous and exogenous ligands. The structure of WbGST also provides a case for the role of the π-cation interaction in stabilizing catalytic Tyr compared to stabilization interactions described for other GSTs. Hence, the obtained information regarding crucial differences in the active sites will support future design of parasite specific inhibitors. Further, the study also evaluates the inhibition of 〈em〉Wb〈/em〉GST and its variants by antifilarial diethylcarbamazine through kinetic assays.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 10 November 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 505, Issue 4〈/p〉 〈p〉Author(s): He-Nan Liu, Nan-Jue Cao, Xun Li, Wei Qian, Xiao-Long Chen〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Diabetic retinopathy (DR) is a progressive microvascular complication associated with diabetes, and remains the leading cause of preventable blindness worldwide. Recent studies have revealed that microRNAs (miRNAs) were involving in the physiological and pathophysiological processes of diabetes and its microvascular and macrovascular complications. The purpose of the current investigation is to identify the candidate miR-211 as a novel biomarker for occurrence and progression of DR in clinical study and experimental research. Firstly, miR-211 was considered as a candidate miRNA identifying by miRNA microarray analysis, Venn diagram analysis, real-time quantitative reverse-transcription polymerase chain reaction (qRT-PCR) and receiver operating characteristic curve in clinical study. Then, the predicted Sirtuin 1 (SIRT1) may be the target gene of miR-211 searching by TargetScan 7.2. Moreover, miR-211 was significantly up-regulated, while SIRT1 mRNA significantly down-regulated measuring by qRT-PCR, meanwhile, SIRT1 protein was significantly down-regulated in coincidence with SIRT1 mRNA detecting by western blot, and even aggravated associated with diabetes duration in diabetic retinal tissues of vivo experiment. Additionally, miR-211 was directly targeted SIRT1 confirming by dual-luciferase reporter assay. Furthermore, with transfection of antagomiR-211, the apoptosis of HUVECs was significantly suppressed employing by flow cytometry analysis, nevertheless the viability of HUVECs was significantly promoted exploiting by Cell Counting Kit-8 assay. Finally, SIRT1 mRNA and SIRT1 protein were significantly up-regulated testing by qRT-PCR and western blot respectively in hyperglycemic HUVECs transfected with antagomiR-211 of vitro experiment. Consequently, the current clinical study and experimental research imply that serum miR-211 as a novel biomarker with high sensitivity and specificity could be associated with occurrence and progression of DR via targeting SIRT1.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 10 November 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 505, Issue 4〈/p〉 〈p〉Author(s): Weiqi Hu, Weimin Xu, Yi Shi, Weijun Dai〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉 〈h6〉Background〈/h6〉 〈p〉Nasopharyngeal carcinoma (NPC) is the most common type of head and neck cancers which is notable for its distinctive pattern of geographical distribution. HOTAIR has been reported to regulate nasopharyngeal carcinoma tumorigenesis and progression. However, the detailed mechanism underlying HOTAIR-promoted nasopharyngeal carcinoma remains not fully understood.〈/p〉 〈/div〉 〈div〉 〈h6〉Methods〈/h6〉 〈p〉We used RT-qPCR approach to examine genes expression and mRNA level. MTT assay and soft agar assay were used to detect cell growth rate in culture and under suspended condition, respectively. Besides, we employed wound healing assay and transwell invasion assay to determine migration and invasion ability of nasopharyngeal carcinoma cells. We predicted direct downstream targets of miR-101 by bioinformatic analysis, which was confirmed by dual luciferase reporter assay.〈/p〉 〈/div〉 〈div〉 〈h6〉Results〈/h6〉 〈p〉HOTAIR was upregulated in NPC tissues and cells. miR-101 inhibitor greatly enhanced HOTAIR knockdown-regulated cell proliferation, migration and invasion of CNE1 and CNE2 cells. miR-101 was shown to directly bind 3′-UTR of COX-2 and downregulate COX-2 expression. Finally, COX-2 overexpression was demonstrated to rescue the tumor phenotypes of nasopharyngeal carcinoma cells attenuated by HOTAIR knockdown or miR-101 mimic.〈/p〉 〈/div〉 〈div〉 〈h6〉Conclusions〈/h6〉 〈p〉Here, we highlight the importance of HOTAIR/miR-101/COX-2 axis in progression of nasopharyngeal carcinoma cells. Our findings provide a novel mechanism for explaining HOTAIR-induced nasopharyngeal carcinoma and help developing the therapeutical strategies by targeting HOTAIR.〈/p〉 〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 10 November 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 505, Issue 4〈/p〉 〈p〉Author(s): Michela Zamboni, Donato Civitareale〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉In non-small lung cancer, the expression of the transcription factor TTF-1/Nkx2.1 correlates with the presence of EGFR mutations, therefore TTF-1/Nkx2.1 expression is used to optimize an EGFR testing strategy and to guide clinical treatment. We investigate the molecular mechanisms underlying the functional connection between EGFR and TTF-1/Nkx2.1 gene expression in lung adenocarcinoma. Using the H1975 cell line as a non-small cell lung cancer model system and short hairpin RNA, we have selected clones with TTF-1/Nkx2.1 silenced expression. We have found that Leucine-rich immunoglobulin repeats-1 (LRIG1) gene is a direct target of TTF-1/Nkx2.1 and the transcription factor binding to the LRIG1 genomic sequence inhibits its gene expression. In TTF-1/Nkx2.1 depleted clones, we have found high levels of LRIG1 and decreased presence of EGFR protein. Furthermore, in TTF-1/Nkx2.1 depleted clones we detected a reduced β-catenin level and we provide experimental evidence indicating that TTF-1/Nkx2.1 gene expression is regulated by β-catenin. Published studies indicate that LRIG1 triggers EGFR degradation and that mutated EGFR induces β-catenin activity. Hence, with the present study we show that mutated EGFR, enhancing β-catenin, stimulates TTF-1/Nkx2.1 gene expression and, at the same time, TTF-1/Nkx2.1, down-regulating LRIG1, sustains EGFR pathway. Therefore, LRIG1 and β-catenin mediate the functional connection between TTF-1/Nkx2.1 and mutated EGFR.〈/p〉〈/div〉 〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0006291X18321570-fx1.jpg" width="500" alt="Image 1" title="Image 1"〉〈/figure〉〈/p〉〈/div〉

Beschreibung: 〈p〉Publication date: 10 November 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 505, Issue 4〈/p〉 〈p〉Author(s): Cong Zu, Guangyuan Qin, Chunshu Yang, Ning Liu, Anning He, Mingdi Zhang, Xinyu Zheng〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉 〈h6〉Purpose〈/h6〉 〈p〉The resistance to 5-FU often limits its clinical effectiveness on breast cancer treatment. Combination therapy thus is employed to overcome this treatment resistance. We here report a potent antitumor effect of Emodin at low dose on chemotherapy sensitivity of MCF-7 breast cancer cells.〈/p〉 〈/div〉 〈div〉 〈h6〉Methods〈/h6〉 〈p〉Cell viability, apoptosis, glutathiones (GSH) concentration and Reactive oxygen species (ROS) activity following Emodin and 5-FU treatment was assessed. Cellular senescence following combined treatment and silence of NRARP was examined by senescence-associated β-galactosidase analysis. Western blot analysis was used to determine changes in the expression of p21, p16, p27, E2F1 and NRARP.〈/p〉 〈/div〉 〈div〉 〈h6〉Results〈/h6〉 〈p〉Low dose Emodin potentiates 5-FU-induced apoptosis of breast cancer cells, in association with inhibition of NRARP, resulting in cellular senescence. RNA interference of NRARP induced cellular senescence in MCF-7 breast cancer cells. Furthermore, the cellular senescence induced by Emodin and 5-FU treatment could be reverted by pcDNA–NRARP.〈/p〉 〈/div〉 〈div〉 〈h6〉Conclusion〈/h6〉 〈p〉These findings provide preclinical evidence for repurposing use of Emodin in combination with chemotherapeutic agents to treat breast cancer as an alternative salvage regimen.〈/p〉 〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 10 November 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 505, Issue 4〈/p〉 〈p〉Author(s): Jae Yeon Choi, Hyun Jin Shin, In Hwa Bae〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Cellular senescence, a distinctive type of irreversible growth arrest, develops in response to various stimuli. Bcl-w, an oncogene and member of the Bcl-2 family, has been reported to promote tumorigenicity in various cancer cells. Here, we sought to explore the potential role of Bcl-w in premature senescence, which has received relatively little research attention. Our findings demonstrate that Bcl-w enhances the activity of senescence-associated β-galactosidase (SA-β-gal) and promotes histone H3 tri-methylation at lysine 9 (H3K9me3) and expressions of p53, Notch2, p21, and p16—hallmarks of the senescent phenotype—in human U251 glioblastoma and H460 lung carcinoma cells. It is also known that microRNAs (miRNAs) regulate processes related to tumor development, such as cell proliferation, differentiation, survival, metabolism, inflammation, invasion, angiogenesis, and senescence. In this context, we found that miR-93-5p inhibited premature cellular senescence by directly suppressing Bcl-w and p21 expressions. Collectively, these findings suggest that targeting miR-93-5p–regulated Bcl-w may be a useful strategy for preventing premature senescence.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 10 November 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 505, Issue 4〈/p〉 〈p〉Author(s): Jong Hyuk Lee, Yoon Hee Chung, Ho Hyun Kim, Joon Seok Bang, Tae Woo Jung, Taekwang Park, Jinwoo Park, Uiseok Kim, Sung Hoon Lee, Ji Hoon Jeong〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉The aim of the present study was to examine the effects of p-coumaric acid on the longitudinal growth of the long bone in adolescent male rats. Teatment with p-coumaric acid significantly increased the tibial length and the height of each growth plate zone and the ratio of 5-bromo-2′-deoxyuridine-positive cells relative to total proliferative cells. Expression of insulin-like growth factor 1 and its receptor in the proliferative and hypertrophic zones, and serum levels of growth hormone and insulin-like growth factor 1 were significantly increased as well in the p-coumaric acid-treated group. Via increasing both the serum level of insulin-like growth factor 1 and its expression, p-coumaric acid could promote cell proliferation in growth plate zones. These results suggest that p-coumaric acid has the potential to increase height and may be a feasible alternative to growth hormone therapy.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: 10 November 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 505, Issue 4〈/p〉 〈p〉Author(s): Xiaoyu Chen, Shan Liu, Wei Zhang, Chunyan Wu, Hanchen Liu, Fang Zhang, Zhongbing Lu, Wenjun Ding〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Recent studies have demonstrated that long-term exposure to fine particulate matter (PM〈sub〉2.5〈/sub〉) increases the risk of central nervous (CNS) diseases. As a basic region-leucine zipper (bZip) transcription factor, nuclear factor erythroid 2-related factor 2 (Nrf2) is essential for protection against chemically induced oxidative stress to restore cellular redox balance. However, the impact of Nrf2 on the neurotoxic effects of PM〈sub〉2.5〈/sub〉 remains to understand. In this study, we exposed wild-type (WT) and Nrf2 knockout (Nrf2〈sup〉−/−〈/sup〉) mice to 1 mg/kg PM〈sub〉2.5〈/sub〉 or deionized water by intranasal instillation for 28 days. After PM〈sub〉2.5〈/sub〉 exposure, Nrf2〈sup〉−/−〈/sup〉 mice exhibited severe nerve injury in olfactory bulb (OB) of mice. In PM〈sub〉2.5〈/sub〉-treated OBs, Nrf2 deficiency resulted in lower levels of antioxidant enzymes, greater induction of oxidative stress, microglia activation, inflammation and nuclear factor kappa B (NF-κB) activation compared to the OBs of WT mice. In PM〈sub〉2.5〈/sub〉-treated BV2 cells, inhibition of Nrf2 activity significantly decreased cell viability and increased the intracellular reactive oxygen species (ROS) generation and nuclear factor kappa B (NF-κB) phosphorylation. Taken together, our results provide the role Nrf2-reuglated antioxidant and cytoprotective enzymes in protective responses to PM〈sub〉2.5〈/sub〉-induced neurotoxicity. Our findings suggest Nrf2-mediated defenses against oxidative stress will help develop new strategies for the prevention and treatment of diseases associated with airborne pollution.〈/p〉〈/div〉 〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0006291X18322034-fx1.jpg" width="500" alt="Image 1" title="Image 1"〉〈/figure〉〈/p〉〈/div〉

Beschreibung: 〈p〉Publication date: Available online 28 March 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Robert A.J. Tucker, Irwin K. Cheah, Barry Halliwell〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Ergothioneine is a biologically important compound that has been shown to be transported by the organic cation transporter novel type 1 (OCTN1). Following this discovery, a variety of alternate functions for OCTN1 have been suggested including an integral function in the extra-neuronal cholinergic system. The present study reaffirms the primacy of ergothioneine over these alternate substrates using natively expressed OCTN1 in HeLa cells. Besides the general transport inhibitors, quinidine, verapamil and pyrilamine no other putative substrate inhibited ergothioneine transport significantly, with only a slight inhibition demonstrated by carnitine. Even compounds structurally similar to ergothioneine failed to inhibit ergothioneine uptake, suggesting high selectivity of OCTN1. Ergothioneine was found to be avidly accumulated even at low concentrations (300 nM) by HeLa cells.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: Available online 28 March 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Yingli Qiao, Xueli Zhang, Guimei Zhao, Zhiheng Liu, Mingyong Yu, Zheping Fang, Xuehua Li〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Although the role of inducible nitric oxide synthase (iNOS) in hepatic ischemia/reperfusion (I/R) injury remains controversial and confusing, with both harmful and beneficial effects in animal studies, the mechanism of these incongruous actions remains unclear. In the current study, we generated bone marrow chimeric mice with hepatocyte-restricted expression of iNOS. Chimeric mice and primary hepatocytes were subjected to I/R or anoxia/reoxygenation stimulation, respectively. The role of iNOS in liver I/R injury and the underlying molecular mechanisms were investigated. Hepatocyte-derived iNOS resulted in hepatoprotection from I/R injury, as well as in vitro experiments. Mechanistically, iNOS upregulates Heat shock protein (HSP) 70 by augmenting heat shock factor 1 (HSF1) binding to the HSP70 gene promoter. Importantly, inhibition of HSP70 partly reversed the iNOS overexpression-mediated hepatoprotection. The present findings demonstrate that hepatocellular iNOS protects from hepatic I/R injury through the HSF1-dependent activation of the HSP70. The upregulation of hepatocellular iNOS may offer a promising strategy for protecting against I/R injury.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: Available online 28 March 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): A.V. Maltsev, E.V. Evdokimovskii, Y.M. Kokoz〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉α2-Adrenoceptors (α2-AR) found in the cardiomyocyte's sarcolemma represent a very important negative feedback for control of myocardial contractility by endogenous catecholamines. Earlier, we showed that the endogenous neurotransmitter agmatine in micromolar concentrations via α2-AR activates the nitric oxide (NO) synthesis, enhancing the Ca〈sup〉2+〈/sup〉 pumping into sarcoplasmic reticulum (SR). In the millimolar doses it inhibits Ca〈sup〉2+〈/sup〉 sequestration by SR Ca〈sup〉2+〈/sup〉 ATPase (SERCA), acting through the first type of imidazoline receptors. Here, we study the functional activity of agmatine, as well as a specific α2-agonist, guanabenz, in respect to spontaneous Ca〈sup〉2+〈/sup〉-transients in SHR cardiomyocytes of the early age (2–2.5 months), and adulthood animals (8–9 months). α2-mediated cardioprotective effect was almost twofold decreased in SHR cardiac cells compared to normotensive rats of the corresponding age, despite the fact that both α2A- and α2B-AR protein levels were significantly increased in SHR cardiomyocytes. NO-mediated facilitation of SERCA activity is substantially reduced in SHR cardiomyocytes vs. normotensive rats. These data suggest that the SHR phenotype starting from early age shows signs of the impaired sarcolemmal α2-AR signaling, which can aggravate the development of this cardiovascular pathology.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: Available online 28 March 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Erika Camacho, Teresa Escalante, Kim Remans, José María Gutiérrez, Alexandra Rucavado〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Abrogation of the hemorrhagic activity of BaP1, a PI Snake Venom Metalloproteinase (SVMP) from the venom of 〈em〉Bothrops asper〈/em〉, was achieved by the substitution of residues in the first part of the Ω loop surrounding the active site by the corresponding residues of a structurally-similar non-hemorrhagic PI SVMP from a related venom. Previous studies by molecular dynamic simulation showed higher flexibility in the first part of the loop in hemorrhagic SVMPs, as compared to non-hemorrhagic SVMPs. It has been suggested that the Ω loop is critical for protein-protein interface and may be involved in the interaction with extracellular matrix proteins, hence influencing the ability of the toxin to bind and hydrolyze basement membrane components. The SVMP with the site mutation completely lost hemorrhagic activity, and only had a partial reduction of proteolytic activity, indicating that this region in the loop plays a key role in the ability to induce hemorrhage. Our findings demonstrate a key structural determinant of the hemorrhagic capacity of PI SVMPs.〈/p〉〈/div〉 〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0006291X19305522-fx1.jpg" width="404" alt="Image 1" title="Image 1"〉〈/figure〉〈/p〉〈/div〉

Beschreibung: 〈p〉Publication date: Available online 28 March 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Yukihiro Baba, Yui Watabe, Hiroshi Sagara, Sumiko Watanabe〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Mammals possess four Sall transcription factors that play various roles in organogenesis. Previously, we found that Sall1 is expressed in microglia in the central nervous system, and it plays pivotal roles in microglia maturation. In the eye, Sall1 was also expressed in the developing lens, and we examined its role in lens development. A knock-in mouse harboring the 〈em〉EGFP〈/em〉 gene in the 〈em〉Sall1〈/em〉 locus (〈em〉Sall1〈/em〉-gfp) was used to analyze the Sall1 expression pattern. In 〈em〉Sall1〈/em〉-gfp/wild, EGFP was expressed throughout the presumptive lens at E11.5, and subsequently the expression in the lens epithelium became weaker. After birth, signals were observed in the equator region. The effects of Sall1 knockout on lens development were examined in 〈em〉Sall1〈/em〉-gfp/gfp. Lens sections revealed small vacuole-like holes and gaps in the center of the lens fibers at E14.5. Subsequently, the vacuoles appeared in most regions of the fiber cells. Electron microscopic analysis indicated that the vacuoles were between the fiber cells, leading to huge gaps. In addition, contact between the lens epithelium and apical end of the fiber cell was disrupted, and there were gaps between the adjoining lens epithelial cells. However, gap junction structure was observed by electron microscopic analysis, and immunostaining of Zo1 showed rather appropriate expression pattern. Immunohistochemistry indicated that the major lens transcription factors Prox1 and Pax6 were expressed in relatively normal patterns. However, although the expression of Prox1 and Pax6 decreased in nuclei in the control lens, it remained in 〈em〉Sall1〈/em〉-gfp/gfp. In addition, lower expression level of c-Maf protein was observed. Therefore, Sall1 is strongly expressed in the lens from the early developmental stage and plays an essential role in the maintenance of fiber cell and lens epithelium adhesion.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: Available online 28 March 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Guido Gessner, Sarah Runge, Michael Koenen, Stefan H. Heinemann, Mascha Koenen, Jan Haas, Benjamin Meder, Dierk Thomas, Hugo A. Katus, Patrick A. Schweizer〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Pathogenic long QT mutations often comprise high phenotypic variability and particularly variants in 〈em〉ANK2 (〈/em〉long QT syndrome 4) frequently lack QT prolongation. We sought to elucidate the genetic and functional background underlying the clinical diversity in a 3-generation family with different cardiac arrhythmias. Next-generation sequencing-based screening of patients with QT prolongation identified the index patient of the family carrying an ANK2-E1813K variant and a previously uncharacterized KCNH2-H562R mutation in a double heterozygous conformation. The patient presented with a severe clinical phenotype including a markedly prolonged QTc interval (544 ms), recurrent syncope due to Torsade de Pointes tachycardias, survived cardiopulmonary resuscitation, progressive cardiac conduction defect, and atrial fibrillation. Evaluation of other family members identified a sister and a niece solely carrying the ANK2-E1813K variant, who showed age-related conduction disease. An asymptomatic second sister solely carried the KCNH2-H562R mutation. Voltage-clamp recordings in 〈em〉Xenopus〈/em〉 oocytes revealed that KCNH2-H562R subunits were non-functional but did not exert dominant-negative effects on wild-type subunits. Expression of KCNH2-H562R in HEK293 cells showed a trafficking deficiency. Co-expression of the C-terminal regulatory domain of ANK2 in 〈em〉Xenopus〈/em〉 oocytes revealed that ANK2-E1813K diminished currents mediated by the combination of wild-type and H562R KCNH2 subunits. Our data suggest that ANK2 functionally interacts with KCNH2 leading to a stronger current suppression and marked aggravation of long QT syndrome in the patient carrying variants in both proteins.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: Available online 28 March 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Yuxue Cao, Lan Xiao, Yanfan Cao, Ashwin Nanda, Chun Xu, Qingsong Ye〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Traditional treatments for bone repair with allografts and autografts are limited by the source of bone substitutes. Bone tissue engineering via a cell-based bone tissue scaffold is a new strategy for treatment against large bone defects with many advantages, such as the accessibility of biomaterials, good biocompatibility and osteoconductivity; however, the inflammatory immune response is still an issue that impacts osteogenesis. Sphingosine 1-phosphate (S1P) is a cell-derived sphingolipid that can mediate cell proliferation, immunoregulation and bone regeneration. We hypothesised that coating S1P on a β-Tricalcium phosphate (β-TCP) scaffold could regulate the immune response and increase osteogenesis. We tested the immunoregulation capability on macrophages and the osteogenic capability on rat bone marrow stromal cells of the coated scaffolds, which showed good biocompatibility. Additionally, the coated scaffolds exhibited dose-dependent inhibition of inflammatory-related gene expression. A high concentration of S1P (0.5 μM) upregulated osteogenic-related gene expression of OPN, OCN and RUNX2, which also significantly increased the alkaline phosphatase activity, as compared with the control group. In conclusion, S1P coated β-TCP scaffold could inhibit inflammation and promote bone regeneration.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: Available online 28 March 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Yurong Wang, Di Guo, Junqiu He, Lihua Song, Huachao Chen, Zhihao Zhang, Ninghuan Tan〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Emerging evidence has shown that metabolism, in particular the synthesis of fatty acids, has great significance for growth and metastasis of colorectal neoplasm. The previous results showed that RA-XII, a natural cyclopeptide isolated from 〈em〉Rubia yunnanensis〈/em〉, inhibits tumor growth and metastasis by AMPK/mTOR/P70S6K pathway and PI3K/AKT/NF-κB pathway. But if or not lipid metabolism involves the antitumor mechanism of RA-XII is not clear. Herein the results indicated that RA-XII reduced the cell motility by decreasing the expressions of β-catenin and β-catenin dependent proteins CD44 and MMP7 in HCT116 cells. Then RA-XII effectively reduced fatty acids levels by decreasing the expression of SREBP-1 and inhibiting the expressions of 〈em〉de novo〈/em〉 fatty acid synthesis proteins FASN and SCD. Moreover the decreased cell motility caused by RA-XII was attenuated with the SREBP-1 knockdown. In addition, the 〈em〉in vivo〈/em〉 experiments also demonstrated that RA-XII inhibited tumor growth and metastasis 〈em〉via〈/em〉 restraining lipogenesis in colorectal neoplasm mouse models. Taken together, these results indicated that RA-XII suppressed the colorectal neoplasm growth and metastasis by inhibition of lipogenesis depended on SREBP-1 suppression.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: Available online 27 March 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Joseph C. Lownik, Daniel H. Conrad, Rebecca K. Martin〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Group 2 innate lymphoid cells (ILC2s) play an important role in the initiation of type-2 immune responses. Numerous targets have been identified that may activate or repress ILC2 function, though few negative regulatory feedback pathways induced upon activation have been shown to be operative in ILC2s. Here we demonstrate that loss of ADAM17 from ILC2s results in a selective defect in IL-33 responsiveness, but not IL-25 responsiveness. We find that IL1R2 is significantly upregulated at both the transcript and protein level in IL-33 activated ILC2s. We are also able to demonstrate that ADAM17 regulates IL1R2 levels on ILC2s in both a constitutive and activation induced manner. Additionally, IL1R2〈sup〉+〈/sup〉 ILC2s, a unique subset of ILC2s, have decreased 〈em〉Il5〈/em〉 and 〈em〉Il13〈/em〉 transcripts following IL-33 stimulation. Overall, these data suggest that the expression of IL1R2 may act as an activation-induced negative regulatory feedback mechanism to decrease ILC2 responsiveness to IL-33.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: Available online 28 March 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Maik Behrens, Ulrike Redel, Kristina Blank, Wolfgang Meyerhof〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉The human sense of taste is devoted to the analysis of the chemical composition of food prior to ingestion. Among the five basic taste qualities bitter taste perception is believed to avoid ingestion of potentially toxic substances. The receptors facilitating the detection of hundreds of chemically different bitter compounds belong to the taste 2 receptor (TAS2R) family, which are part of the G protein-coupled superfamily. Although the chemical classes of bitter compounds that have been identified as agonists of one of the 25 potentially functional human bitter taste receptors cover an enormous chemical space, one distinct group of bitter compounds, the bitter salts have not been assigned to any bitter taste receptor. To close this gap, we screened the entire human bitter taste receptor repertoire by functional calcium mobilization assays with the most famous bitter salt, magnesium sulfate, also known as Epsom salt. Although the profound pharmacological activity and the bitter taste of spring water containing magnesium sulfate has been known since 1697, the molecular basis for its taste has not been elucidated until now. Our screening resulted in the identification of a single receptor, the TAS2R7, responding to magnesium sulfate at concentrations humans perceive this salt as bitter. Subsequently, TAS2R7 was stimulated with other salts and it was found that this receptor also responds to manganese〈sup〉2+〈/sup〉 and iron〈sup〉2+〈/sup〉 ions, but not to potassium ions. Magnesium sulfate is known to exert a number of beneficial effects on the human body and thus, has been used as medicine against premature uterine contractions, as anti-arrhythmic drug and as laxative, however, magnesium sulfate overdosage can result in cardiac arrest and thus have fatal consequences. Therefore, it appears reasonable that nature placed TAS2R7 as sentinel for high concentrations of bitter salts on our tongues.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: Available online 25 March 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Shasha Yang, Jie Tian, Fufeng Zhang, Aibin Liu, Bin Xie, Qiong Chen〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Lung ischemia-reperfusion injury (LIRI) often results in respiratory insufficiency after pulmonary embolism, lung transplantation, etc. To investigate the role of HSP22 in LIRI mice, ischemia-reperfusion injury was established in the left lung of an HSP22 overexpression transgenic mouse. Twelve HSP22 transgenic (TG) mice and twelve wild-type (WT) mice were randomly divided into 2 groups: the sham-operated group (SO: TG-SO, WT-SO) and the ischemia-reperfusion group (I/R: TG-I/R, WT-I/R), respectively. We tested the PaO〈sub〉2〈/sub〉, W/D ratio, and MDA level; observed morphology changes; and calculated the index of alveolar damage. HSP22 expression was examined in lung tissues of TG and WT C57BL mice by immunohistochemistry. TUNEL assay was performed to measure apoptosis. We found that HSP22 was significantly overexpressed in TG mice. There was no difference in PaO〈sub〉2〈/sub〉 among the four groups. In the I/R group, the W/D ratio, MDA and index of alveolar damage were higher than those in the SO group. Moreover, compared with WT-I/R group, the W/D ratio, MDA and index of alveolar damage in the TG-I/R group were significantly decreased. Apoptosis in the I/R groups was increased compared to that in the SO groups, while apoptosis in the TG-I/R groups was decreased compared to that in the WT-I/R groups. Our results showed that HSP22 TG mice and the LIRI model were successfully established. In addition, HSP22 overexpression has protective effects on LIRI by inhibiting lipid peroxidation and apoptosis.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: Available online 25 March 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Wei Yu, Huaye Zhang, Mi Ryung Shin, Federico Sesti〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉 〈p〉Voltage-gated potassium (K〈sup〉+〈/sup〉) channel sub-family B member 1 (KCNB1, Kv2.1) is known to undergo oxidation-induced oligomerization during aging but whether this process affects brain's physiology was not known. Here, we used 10, 16 and 22 month-old transgenic mice overexpressing a KCNB1 variant that does not oligomerize (Tg-C73A) and as control, mice overexpressing the wild type (Tg-WT) channel and non-transgenic (non-Tg) mice to elucidate the effects of channel's oxidation on cognitive function. Aging mice in which KCNB1 oligomerization is negligible (Tg-C73A), performed significantly better in the Morris Water Maze (MWM) test of working memory compared to non-Tg or Tg-WT mice. KCNB1 and synapsin-1 co-immunoprecipitated and the cognitive impairment in the MWM was associated with moderate loss of synapsin-1 in pre-synaptic structures of the hippocampus, whereas neurodegeneration and neuronal loss were not significantly different in the various genotypes.〈/p〉 〈p〉We conclude that moderate oxidation of the KCNB1 channel during aging can influence neuronal networks by affecting synaptic function.〈/p〉 〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: Available online 25 March 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Alexandra Woychek, Kristina Kligys, Susan B. Hopkinson, Jonathan C.R. Jones〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉The α6β4 integrin heterodimer is an essential component of hemidesmosomes (HDs) and HD-related structures, which adhere epithelial cells to the underlying extracellular matrix. In this study, we focused on the importance of the α6 integrin 3′ untranslated region (UTR) in α6β4 integrin localization. To do so, A549 cells (a type II lung alveolar cell line) and immortalized human epidermal keratinocytes (iHEK) were infected with adenovirus encoding the entire α6 integrin protein with or without portions of its 3′UTR. In infected A549 cells, we detected α6β4 integrin heterodimers containing the product of the adenovirus, regardless of whether the α6 integrin 3′UTR was present. However, only those α6 integrin proteins whose messages contained bases 4770–5633 of the α6 integrin 3′UTR were targeted to matrix adhesion sites. Moreover, overexpression of the full length α6 integrin 3′UTR, minus the coding sequence, in A549 cells disrupts the localization of endogenous α6β4 integrin heterodimers. Following infection of iHEKs with the same adenovirus, the induced α6 integrin protein localizes to HDs regardless of whether its message possessed a 3′UTR. In sharp contrast, in α6 integrin depleted iHEKs, restoring α6 integrin expression using the coding sequence alone via adenoviral transduction resulted in α6 integrin preferentially forming α6β1 rather than α6β4 integrin heterodimers. α6β4 integrin was only observed in knocked down cells following infection of adenovirus encoding the α6 integrin coding sequence with its 3′UTR. In summary, our data indicate that the α6 integrin 3′UTR is a key regulator of α6β4 integrin heterodimer assembly and incorporation at sites of cell-matrix adhesion.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: Available online 29 March 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Shixin Zhang, Yan Liang, Yuanyuan Wu, Xuedan Chen, Kai Wang, Juan Li, Xingying Guan, Gang Xiong, Kang Yang, Yun Bai〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉An increasing number of long noncoding RNAs (lncRNAs) have been discovered, and dysregulation of lncRNAs plays critical roles in tumorigenesis and tumor progression. In this study, we identified a novel lncRNA LINC01980, located in both the cytoplasm and nucleus, which was significantly upregulated in esophageal squamous cell carcinoma (ESCC) tissues through microarray profiling. Further analysis revealed that LINC01980 overexpression was positively correlated with deeper invasion of cancer, positive lymph node metastasis, and advanced TNM stage. Additionally, high LINC01980 expression in ESCC tissues was associated with poor prognosis. 〈em〉In vitro〈/em〉 and 〈em〉in vivo〈/em〉 experiments demonstrated that LINC01980 promoted ESCC growth. EdU incorporation assay implied that LINC01980 accelerated ESCC proliferation. Flow cytometry analysis showed that knockdown of LINC01980 induced cell cycle arrest and increased apoptosis. Microarray analysis indicated that LINC01980 upregulated the expression of growth arrest and DNA damage inducible 45 alpha (GADD45A). Further experiments demonstrated that GADD45A promoted ESCC cell growth, indicating that GADD45A may be a downstream target of LINC01980. In conclusion, this study identified LINC01980 as a novel potential oncogene in ESCC, which can be a promising biomarker for prognosis and therapeutic targeting in ESCC.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: Available online 30 March 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Sho Wagai, Atsushi Kasamatsu, Manabu Iyoda, Fumihiko Hayashi, Kazuya Hiroshima, Shusaku Yoshimura, Isao Miyamoto, Dai Nakashima, Yosuke Endo-Sakamoto, Masashi Shiiba, Hideki Tanzawa, Katsuhiro Uzawa〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Unc-93 homolog B1 (UNC93B1), a transmembrane protein, is correlated with immune diseases, such as influenza, herpes simplex encephalitis, and the pathogenesis of systemic lupus erythematosus; however, the role of UNC93B1 in cancers including human oral squamous cell carcinomas (OSCCs) remains unknown. In the current study, we investigated the UNC93B1expression level in OSCCs using quantitative reverse transcription-polymerase chain reaction, immunoblot analysis, and immunohistochemistry. Our data showed that UNC93B1 mRNA and protein expressions increased markedly (p 〈 0.05) in OSCCs compared with normal cells and tissues and that high expression of UNC93B1 in OSCCs was related closely to tumoral size. UNC93B1 knockdown (shUNC93B1) OSCC cells showed decreased cellular proliferation by cell-cycle arrest in the G1 phase with up-regulation of p21〈sup〉Cip1〈/sup〉 and down-regulation of CDK4, CDK6, cyclin D1, and cyclin E. We also found that granulocyte macrophage colony-stimulating factor (GM-CSF) was down-regulated significantly (p 〈 0.05) in shUNC93B1 OSCC cells. Moreover, inactivation of GM-CSF using neutralization antibody led to cell-cycle arrest at the G1 phase similar to the phenotype of the shUNC93B1 cells. The current findings indicated that UNC93B1 might play a crucial role in OSCC by controlling the secretion level of GM-CSF involved in tumoral growth and could be a potential therapeutic target for OSCCs.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: Available online 29 March 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Seungho Choi, Hyun Jin Jung, Min Woo Kim, Ju-Hee Kang, Dongyun Shin, Yeong-Su Jang, Yeo Sung Yoon, Seung Hyun Oh〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Liver fibrosis is characterized by formation of scar tissue in the liver. The role of STAT3 signaling has been implicated on activating hepatic stellate cells (HSC) to myofibroblast-like cells in liver fibrosis. Major factors that activate STAT3 signaling are TGF-β1 and IL-6, which are upregulated in the liver in patients afflicted with liver fibrosis. Recent reports indicate that not only IL-6, but also the non-canonical signaling pathway of TGF-β1 is associated with STAT3 signaling. In this study, we demonstrate a new function of the STAT3 inhibitor, STX-0119, in liver fibrosis. STX-0119 is an inhibitor of STAT3 dimerization, which is required for nuclear localization of STAT3. We first investigated the anti-fibrotic effect of STX-0119 in 〈em〉in vitro〈/em〉 experiments. Exposure to STX-0119 inhibited the nuclear localization of STAT3 in HSCs, resulting in decreased expression of its target genes, such as col1a1 and αSMA. In addition, STX-0119 also inhibited the TGF-β1/IL-6-induced activation of HSCs. Next, we examined the 〈em〉in vivo〈/em〉 effect of STX-0119 in the liver fibrosis mouse model using thioacetamide (TAA) and carbon tetrachloride (CCl〈sub〉4〈/sub〉). STX-0119 attenuated the TAA-induced liver fibrosis by inhibiting activation of HSCs to myofibroblast-like cells. Consistent with the 〈em〉in vivo〈/em〉 results using TAA-induced liver fibrosis model, treatment of STX-0119 similarly attenuated CCl〈sub〉4〈/sub〉-induced liver fibrosis. In conclusion, we believe that STX-0119 inhibits the development of liver fibrosis by blocking the activation of hepatic stellate cells. These results indicate that STX-0119 is a potential new therapeutic strategy to prevent disease progression to cirrhosis.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: Available online 30 March 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Yao Liu, Tongyu Zhang, Yanbin Wang, Pei Wu, Yuchen Li, Chunlei Wang, Shancai Xu, Huaizhang Shi〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Apelin, an endogenous ligand for the orphan G-protein-coupled receptor APJ, possesses anti-apoptotic and neuroprotective properties. It has been shown to be a protective factor for different types of central nervous system insults, such as ischemia and traumatic brain injury. Here, we investigated the effects of apelin-13 on early brain injury (EBI) following subarachnoid hemorrhage (SAH), and the underlying molecular mechanisms involved. Apelin-13 was delivered to rats via intracerebroventricular administration. Neurological scores, brain water content and neuronal apoptosis were measured 24 h after SAH. The PI3K/Akt inhibitor LY294002 or GLP-1R siRNA were injected into the lateral cerebral ventricle before induction of SAH. Changes in the expression of p-Akt, GLP-1R and apoptosis-associated proteins (Bax, Bcl-2, cleaved caspase-3) were then investigated. Results showed that the levels of GLP-1R in neurons increased significantly, reaching a peak at 24 h after the induction of SAH. Treatment with apelin-13 improved neurological deficits, as well as alleviated brain edema and apoptotic cell death. Apelin-13 was also able to increase the levels of p-Akt, GLP-1R and Bcl-2, while inhibiting the expression levels of Bax and cleaved caspase-3. The anti-apoptotic and neuroprotective effects of apelin-13 were partially reversed by addition of LY294002 or GLP-1R siRNA. These results provide evidence that apelin-13 attenuates EBI following SAH via suppressing neuronal apoptosis, and that this effect may act partially via the activation of the GLP-1R/PI3K/Akt signaling pathway.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: Available online 29 March 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Liqiang Qiu, Changwu Xu, Jing Chen, Qi Li, Hong Jiang〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉P300/CBP-associated factor (PCAF) regulates vascular inflammation. This study was to explore the effect of PCAF on the proliferation and migrationof vascular smooth muscle cells (VSMCs) and neointimal hyperplasia in balloon-injured rat carotid artery. Downregulation of PCAF remarkably suppressed VSMCs proliferation and migration induced by lipopolysaccharide, and also significantly inhibit the nuclear translocation of nuclear factor-kappaB p65. Meanwhile, downregulation of PCAF inhibited the mRNA expression of tumor necrosis factor-α and interleukin-6, and also the levels in culture supernatants. Moreover, downregulation of PCAF profoundly reduced the intima area and the ratio of intima area to media area in balloon-injured rat carotid artery. In addition, the expression of PCNA and NF-κB p65 in intima were decreased by downregulation of PCAF. These results highlight that PCAF may be a potential target for prevention and treatment of neointimal hyperplasia and restenosis after angioplasty.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: Available online 30 March 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Wenwen Fu, Guannan Meng, Xiaomeng Yang, Lilei Yu, Hong Jiang〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉After myocardial infarction (MI), increased platelet number and size are inversely related to the outcomes of patients. Our previous study confirmed an excessive thrombopoiesis taking place in the bone marrow after MI. However, the mechanisms remain unknown. It has been reported that the sympathetic stimulation by noise or exercise can promote megakaryocyte (MK) producing platelets which is mediated by α2-adrenoceptor. Here, using whole-mount staining combined with western blotting and ELISA assay, we vividly showed an activation of the bone marrow sympathetic nervous system (SNS) after MI. Interestingly, we observed a direct spatial attachment between MKs and the sympathetic nerves. The administration of α-adrenoceptor antagonist, phentolamine or prazosin, could effectively attenuate post-MI MK cellularity and maturity, and alter the distribution of MK away from the bone marrow vessels. Surprisingly, the antagonists did not suppress the final stage of platelet formation. MI mice treated with phentolamine or prazosin showed elevating circulating platelets comparable as those treated with PBS as the control. Together, this study demonstrated that the activation of bone marrow SNS after MI regulates megakaryocyte expansion but not platelet production. Therefore, targeting sympathetic activation might become a novel approach for controlling post-MI bone marrow MK development, but other approaches are still needed to effectively reduce the platelet numbers.〈/p〉〈/div〉 〈/div〉

Beschreibung: 〈p〉Publication date: Available online 30 March 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Noriyoshi Yoshinaga, Keisuke Shindo, Yusuke Matsui, Yoko Takiuchi, Hirofumi Fukuda, Kayoko Nagata, Kotaro Shirakawa, Masayuki Kobayashi, Shunichi Takeda, Akifumi Takaori-Kondo〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Host DNA damage response molecules affect retroviral infection, as DNA intermediates of the viruses play essential roles in the viral life cycles. Although several such molecules have been reported, interactions between HIV-1 and host DNA damage response molecules have not been fully elucidated. To screen DNA damage response molecules that might affect HIV-1 infection, a set of 32 DNA-repair-deficient DT40 isogenic mutant cells were tested for HIV-1 infectivity. Seven out of the 32 clones showed less than 50% infectivity compared to parental DT40 cells, implying that DNA repair molecules deficient in these cells might support HIV-1 infection. Of these, 〈em〉EXO1 −/−, TP53BP1 −/−〈/em〉 and 〈em〉WRN −/−〈/em〉 cells showed more than twofold accumulation of two long terminal repeat circles and less than 50% integrated proviral DNA in quantitative-PCR analyses, indicating that the integration step is impaired. 〈em〉RAD18 −/−〈/em〉 cells showed twofold higher HIV-1 infectivity and increased reverse transcription products at earlier time points, suggesting that RAD18 suppresses reverse transcription. The HIV-1 suppressive effects of RAD18 were confirmed by over-expression and knockdown experiments in human cells. L274P, a DNA-binding-impaired mutant of RAD18, showed impaired HIV-1 suppression and DNA binding, suggesting that binding HIV-1 DNA intermediates is critical for RAD18 to suppress reverse transcription and HIV-1 infection. Our data help understand interactions between host DNA damage response molecules and viral DNA.〈/p〉〈/div〉 〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0006291X19305686-fx1.jpg" width="282" alt="Image 1" title="Image 1"〉〈/figure〉〈/p〉〈/div〉