bluuebunny/crossref_metadata_2025 huggingface dataset:
PDI-mediated S-nitrosylation of DRP1 facilitates DRP1-S616 phosphorylation and mitochondrial fission in CA1 neurons
Authors: Lee Duk-shin , Kim Ji-Eun
Published: August 2018

Dynamin-related protein 1 (DRP1) is a key molecule to regulate mitochondrial fission. DRP1 activity is modulated by phosphorylation and S-nitrosylation on serine and cysteine residues, respectively. However, it is still unexplored whether S-nitrosylation of DRP1 affects its phosphorylation. In the present study, we found that Nω-nitro-l-arginine methyl ester hydrochloride (l-NAME, a NOS inhibitor) abolished S-nitrosylated (SNO-DRP1) and DRP1-serine (S) 616 phosphorylation levels in CA1 neurons under physiological condition. l-NAME led to mitochondrial elongation. In spite of the sustained NO synthesis, status epilepticus (a prolonged seizure activity, SE) diminished SNO-DRP1 and DRP1-S616 levels in CA1 neurons, accompanied by the reduced protein disulfide isomerase (PDI) expression and mitochondrial elongation. SE did not influence thioredoxin 1 (Trx1, a denitrosylating enzyme) activity, which was unaffected by l-NAME under physiological and post-SE condition. PDI knockdown decreased SNO-DRP1 and DRP1-S616 levels concomitant with mitochondrial elongation in CA1 neurons without altered NO synthesis under physiological condition. These findings indicate that PDI may be a NO donor of DRP1 to regulate DRP1-S616 phosphorylation, independent of Trx1 activity. Therefore, we suggest that PDI-mediated S-nitrosylation of DRP1 may be one of the major regulatory modifications for mitochondrial dynamics.
Rab37 mediates exocytosis of secreted frizzled-related protein 1 to inhibit Wnt signaling and thus suppress lung cancer stemness
Authors: Cho Shu-Huei , Kuo I-Ying , Lu Pei-Jung Frank , Tzeng Hong-Tai , Lai Wu-Wei , Su Wu-Chou , Wang Yi-Ching
Published: August 2018

Recent studies have revealed that dysregulated Rab small GTPase-mediated vesicle trafficking pathways are associated with cancer progression. However, whether any of the Rabs plays a suppressor role in cancer stemness is least explored. Rab37 has been postulated as a tumor suppressive small GTPase for trafficking anti-tumor cargos. Here, we report a previously uncharacterized mechanism by which Rab37 mediates exocytosis of secreted frizzled-related protein-1 (SFRP1), an extracellular antagonist of Wnt, to suppress Wnt signaling and cancer stemness in vitro and in vivo. Reconstitution experiments indicate that SFRP1 secretion is crucial for Rab37-mediated cancer stemness suppression and treatment with SRPP1 recombinant protein reduces xenograft tumor initiation ability. Clinical results confirm that concordantly low Rab37, low SFRP1, and high Oct4 stemness protein expression profile can be used as a biomarker to predict poor prognosis in lung cancer patients. Our findings reveal that Rab37-mediated SFRP1 secretion suppresses cancer stemness, and dysregulated Rab37-SFRP1 pathway confers cancer stemness via the activation of Wnt signaling. Rab37-SFRP1-Wnt axis could be a potential therapeutic target for attenuating lung cancer stemness.
HGF-mediated crosstalk between cancer-associated fibroblasts and MET-unamplified gastric cancer cells activates coordinated tumorigenesis and metastasis
Authors: Ding Xusheng , Ji Jun , Jiang Jinling , Cai Qu , Wang Chao , Shi Min , Yu Yingyan , Zhu Zhenggang , Zhang Jun
Published: August 2018

Cancer-associated fibroblasts (CAFs) are important components of tumor stroma and play a key role in tumor progression. CAFs involve in crosstalk with tumor cells through various kinds of cytokines. In the present study, we screened hepatocyte growth factor (HGF) as a cytokine predominantly originating from CAFs. CAFs-derived HGF was found to promote MET-unamplified gastric cancer (GC) proliferation, migration, and invasion through the activation of HGF/c-Met/STAT3/twist1 pathway. It also activated interleukin (IL)-6/IL-6R/JAK2/STAT3/twist1 pathway by up-regulating IL-6R expression. As IL-6 was also found to upregulate c-Met expression, we identified the cooperation of HGF and IL-6 in enhancing the characteristics of CAFs. In vivo experiments revealed that CAFs-derived HGF promoted tumorigenesis and metastasis of MET-unamplified GC. Gene set enrichment analysis (GSEA) was performed to confirm our findings. Our study found that the increased expression of HGF in CAFs induced by MET-unamplified GC contributed to the malignant phenotype of both MET-unamplified GC and CAFs in tumor microenvironment.
Zinc induces CDK5 activation and neuronal death through CDK5-Tyr15 phosphorylation in ischemic stroke
Authors: Tuo Qing-Zhang , Liuyang Zhen-Yu , Lei Peng , Yan Xiong , Shentu Yang-Ping , Liang Jia-Wei , Zhou Huan , Pei Lei , Xiong Yan , Hou Tong-Yao , Zhou Xin-Wen , Wang Qun , Wang Jian-Zhi , Wang Xiao-Chuan , Liu Rong
Published: August 2018

CDK5 activation promotes ischemic neuronal death in stroke, with the recognized activation mechanism being calpain-dependent p35 cleavage to p25. Here we reported that CDK5-Tyr15 phosphorylation by zinc induced CDK5 activation in brain ischemic injury. CDK5 activation and CDK5-Tyr15 phosphorylation were observed in the hippocampus of the rats that had been subjected to middle cerebral artery occlusion, both of which were reversed by pretreatment with zinc chelator; while p35 cleavage and calpain activation in ischemia were not reversed. Zinc incubation resulted in CDK5-Tyr15 phosphorylation and CDK5 activation, without increasing p35 cleavage in cultured cells. Site mutation experiment confirmed that zinc-induced CDK5 activation was dependent on Tyr15 phosphorylation. Further exploration showed that Src kinase contributed to zinc-induced Tyr15 phosphorylation and CDK5 activation. Src kinase inhibition or expression of an unphosphorylable mutant Y15F-CDK5 abolished Tyr15 phosphorylation, prevented CDK5 activation and protected hippocampal neurons from ischemic insult in rats. We conclude that zinc-induced CDK5-Tyr15 phosphorylation underlies CDK5 activation and promotes ischemic neuronal death in stroke.
Kaempferol induces autophagic cell death via IRE1-JNK-CHOP pathway and inhibition of G9a in gastric cancer cells
Authors: Kim Tae Woo , Lee Seon Young , Kim Mia , Cheon Chunhoo , Ko Seong-Gyu
Published: August 2018

Kaempferol, a flavonoid, found in traditional medicine, fruits, and vegetables, and an HDAC inhibitor, is a powerful anti-cancer reagent against various cancer cell lines. However, detailed mechanisms involved in the treatment of gastric cancer (GC) using kaempferol are not fully understood. In our study, we investigated the biological activity and molecular mechanism involved in kaempferol-mediated treatment of GC. Kaempferol promoted autophagy and cell death, and increased LC3-I to LC3-II conversion and the downregulation of p62 in GC. Furthermore, our results showed that kaempferol induces autophagic cell death via the activation of the IRE1-JNK-CHOP signaling, indicating ER stress response. Indeed, the inhibition of ER stress suppressed kaempferol-induced autophagy and conferred prolonged cell survival, indicating autophagic cell death. We further showed that kaempferol mediates epigenetic change via the inhibition of G9a (HDAC/G9a axis) and also activates autophagic cell death. Taken together, our findings indicate that kaempferol activates the IRE1-JNK-CHOP signaling from cytosol to nucleus, and G9a inhibition activates autophagic cell death in GC cells.
Presenilin 1 deficiency suppresses autophagy in human neural stem cells through reducing γ-secretase-independent ERK/CREB signaling
Authors: Chong Cheong-Meng , Ke Minjing , Tan Yuan , Huang Zhijian , Zhang Ke , Ai Nana , Ge Wei , Qin Dajiang , Lu Jia-Hong , Su Huanxing
Published: August 2018

Autophagy impairment is commonly implicated in the pathological characteristic of Alzheimer’s disease (AD). Presenilin 1 (PS1) expression in human brain gradually decreases with age and its mutations account for the most common cases of early-onset familial Alzheimer’s disease (FAD). The dominant autophagy phenotypes occur in PS1-knockout and PS1 mutant neurons; it is still unknown whether PS1 deficiency causes serious autophagy impairment in neural stem cells (NSCs). Herein, we generated the heterozygote and homozygote of PS1 knockout in human induced pluripotent stem cells (iPSCs) via CRISPR/Cas9-based gene editing and differentiated them into human NSCs. In these human PS1-deficient NSCs, reduced autophagosome formation and downregulated expression of autophagy–lysosome pathway (ALP)-related mRNAs, as well as proteins were observed. Mechanistically, ERK/CREB inhibition and GSK3β activation had key roles in reducing TFEB expression in PS1-knockout NSCs. Pharmacological inhibition of GSK3β upregulated the expression of TFEB and ALP-related proteins in PS1-knockout NSCs, whereas this effect could be blocked by CREB inhibition. These findings demonstrate that PS1 deficiency causes autophagy suppression in human NSCs via downregulating ERK/CREB signaling.
The activation of microRNA-520h–associated TGF-β1/c-Myb/Smad7 axis promotes epithelial ovarian cancer progression
Authors: Zhang Jing , Liu Wenxue , Shen Fangqian , Ma Xiaoling , Liu Xiaorui , Tian Fuju , Zeng Weihong , Xi Xiaowei , Lin Yi
Published: August 2018

Among the gynaecological cancers, epithelial ovarian cancer (EOC) has the highest lethality because of the high incidence of tumour progression and metastasis. Exploration of the detailed mechanisms underlying EOC metastasis and the identification of crucial targets is important to better estimate the prognosis and improve the treatment of this disease. The present study aimed to identify the role of miR-520h in the prognosis of patients with EOC, and the mechanisms of its involvement in EOC progression. We showed that miR-520h was upregulated in 116 patients with EOC, especially in those with advanced-stage disease, and high miR-520h expression predicted poor outcome. Furthermore, ectopic expression of miR-520h enhanced EOC cell proliferation, migration and invasion, and induced epithelial–mesenchymal transition in vitro and in vivo. miR-520h promoted EOC progression by downregulating Smad7, and subsequently activating the TGF-β signalling pathway. Most importantly, TGF-β1 stimulation increased miR-520h expression in EOC cells by upregulating its transcription factor c-Myb. In conclusion, we described the role of the TGF-β1/c-Myb/miR-520h/Smad7 axis in EOC metastasis, and highlighted the possible use of miR-520h as a prognostic marker for EOC.
Structural and biochemical insights into small RNA 3′ end trimming by Arabidopsis SDN1
Authors: Chen Jiayi , Liu Li , You Chenjiang , Gu Jiaqi , Ruan Wenjie , Zhang Lu , Gan Jianhua , Cao Chunyang , Huang Ying , Chen Xuemei , Ma Jinbiao
Published: August 2018

A family of DEDDh 3′→5′ exonucleases known as Small RNA Degrading Nucleases (SDNs) initiates the turnover of ARGONAUTE1 (AGO1)-bound microRNAs in Arabidopsis by trimming their 3′ ends. Here, we report the crystal structure of Arabidopsis SDN1 (residues 2-300) in complex with a 9 nucleotide single-stranded RNA substrate, revealing that the DEDDh domain forms rigid interactions with the N-terminal domain and binds 4 nucleotides from the 3′ end of the RNA via its catalytic pocket. Structural and biochemical results suggest that the SDN1 C-terminal domain adopts an RNA Recognition Motif (RRM) fold and is critical for substrate binding and enzymatic processivity of SDN1. In addition, SDN1 interacts with the AGO1 PAZ domain in an RNA-independent manner in vitro, enabling it to act on AGO1-bound microRNAs. These extensive structural and biochemical studies may shed light on a common 3′ end trimming mechanism for 3′→5′ exonucleases in the metabolism of small non-coding RNAs.
Exploring Peltier effect in organic thermoelectric films
Authors: Jin Wenlong , Liu Liyao , Yang Tao , Shen Hongguang , Zhu Jia , Xu Wei , Li Shuzhou , Li Qing , Chi Lifeng , Di Chong-an , Zhu Daoben
Published: August 2018

Organic materials are emerging thermoelectric candidates for flexible power generation and solid-cooling applications. Although the Peltier effect is a fundamental thermoelectric effect that enables site-specific and on-demand cooling applications, the Peltier effect in organic thermoelectric films have not been investigated. Here we experimentally observed and quasi-quantitatively evaluated the Peltier effect in a poly(Ni-ett) film through the fabrication of thermally suspended devices combined with an infrared imaging technique. The experimental and simulation results confirm effective extraction of the Peltier effect and verify the Thomson relations in organic materials. More importantly, the working device based on poly(Ni-ett) film yields maximum temperature differences as large as 41 K at the two contacts and a cooling of 0.2 K even under heat-insulated condition. This exploration of the Peltier effect in organic thermoelectric films predicts that organic materials hold the ultimate potential to enable flexible solid-cooling applications.
Genomic and expression profiling reveal molecular heterogeneity of disseminated tumor cells in bone marrow of early breast cancer
Authors: Magbanua Mark Jesus M. , Rugo Hope S. , Hauranieh Louai , Roy Ritu , Scott Janet H. , Lee Jen Chieh , Hsiao Feng , Sosa Eduardo V. , van’t Veer Laura , Esserman Laura J. , Park John W.
Published: August 2018

Detection of disseminated tumor cells (DTCs) in bone marrow is an established negative prognostic factor. We isolated small pools of (~20) EPCAM-positive DTCs from early breast cancer patients for genomic profiling. Genome-wide copy number profiles of DTC pools (n = 45) appeared less aberrant than the corresponding primary tumors (PT, n = 16). PIK3CA mutations were detected in 26% of DTC pools (n = 53), none of them were shared with matched PTs. Expression profiling of DTC pools (n = 30) confirmed the upregulation of EPCAM expression and certain oncogenes (e.g., MYC and CCNE1), as well as the absence of hematopoietic features. Two expression subtypes were observed: (1) luminal with dual epithelial–mesenchymal properties (high ESR1 and VIM/CAV1 expression), and (2) basal-like with proliferative/stem cell-like phenotype (low ESR1 and high MKI67/ALDH1A1 expression). We observed high discordance between ESR1 (40%) and ERRB2 (43%) expression in DTC pools vs. the clinical ER and HER2 status of the corresponding primary tumors, suggesting plasticity of biomarker status during dissemination to the bone marrow. Comparison of expression profiles of DTC pools with available data from circulating tumor cells (CTCs) of metastatic breast cancer patients revealed gene expression signatures in DTCs that were unique from those of CTCs. For example, ALDH1A1, CAV1, and VIM were upregulated in DTC pools relative to CTCs. Taken together, analysis of pooled DTCs revealed molecular heterogeneity, possible genetic divergence from corresponding primary tumor, and two distinct subpopulations. Validation in larger cohorts is needed to confirm the presence of these molecular subtypes and to evaluate their biological and clinical significance.
Correlating nuclear morphometric patterns with estrogen receptor status in breast cancer pathologic specimens
Authors: Rawat Rishi R. , Ruderman Daniel , Macklin Paul , Rimm David L. , Agus David B.
Published: August 2018

In this pilot study, we introduce a machine learning framework to identify relationships between cancer tissue morphology and hormone receptor pathway activation in breast cancer pathology hematoxylin and eosin (H&E)-stained samples. As a proof-of-concept, we focus on predicting clinical estrogen receptor (ER) status—defined as greater than one percent of cells positive for estrogen receptor by immunohistochemistry staining—from spatial arrangement of nuclear features. Our learning pipeline segments nuclei from H&E images, extracts their position, shape and orientation descriptors, and then passes them to a deep neural network to predict ER status. After training on 57 tissue cores of invasive ductal carcinoma (IDC), our pipeline predicted ER status in an independent test set of patient samples (AUC ROC = 0.72, 95%CI = 0.55–0.89, n = 56). This proof of concept shows that machine-derived descriptors of morphologic histology patterns can be correlated to signaling pathway status. Unlike other deep learning approaches to pathology, our system uses deep neural networks to learn spatial relationships between pre-defined biological features, which improves the interpretability of the system and sheds light on the features the neural network uses to predict ER status. Future studies will correlate morphometry to quantitative measures of estrogen receptor status and, ultimately response to hormonal therapy.
An external quantum efficiency of >20% from solution-processed poly(dendrimer) organic light-emitting diodes
Authors: Maasoumi Fatemeh , Jansen-van Vuuren Ross D. , Shaw Paul E. , Puttock Emma V. , Nagiri Ravi Chandra Raju , McEwan Jake A. , Bown Mark , O’Connell Jenny L. , Dunn Christopher J. , Burn Paul L. , Namdas Ebinazar B.
Published: August 2018

Controlling the orientation of the emissive dipole has led to a renaissance of organic light-emitting diode (OLED) research, with external quantum efficiencies (EQEs) of >30% being reported for phosphorescent emitters. These highly efficient OLEDs are generally manufactured using evaporative methods and are comprised of small-molecule heteroleptic phosphorescent iridium(III) complexes blended with a host and additional layers to balance charge injection and transport. Large area OLEDs for lighting and display applications would benefit from low-cost solution processing, provided that high EQEs could be achieved. Here, we show that poly(dendrimer)s consisting of a non-conjugated polymer backbone with iridium(III) complexes forming the cores of first-generation dendrimer side chains can be co-deposited with a host by solution processing to give highly efficient devices. Simple bilayer devices comprising the emissive layer and an electron transport layer gave an EQE of >20% at luminances of up to ≈300 cd/m2, showing that polymer engineering can enable alignment of the emissive dipole of solution-processed phosphorescent materials.
Role of proteins in the degradation of relatively inert alloys in the human body
Authors: Hedberg Yolanda S.
Published: August 2018

Many biomedical materials used today for applications such as orthopedic, dental, and cardiovascular implants and devices are made of corrosion-resistant, ‘inert’, metallic materials of the cobalt–chromium, titanium, and stainless steel alloy groups. This perspective focuses on the role of proteins in the degradation of these materials in a human body environment. After adsorption, the proteins interact relatively slowly with the metal and metal surface oxide. A number of factors, including the individual body chemistry (especially the presence of inflammatory cells producing oxidative species), determine whether the proteins can bind to metals in the surface oxide and whether the metal–protein conjugates can detach from the surface. Metals in the forms of protein-bound metal ions or nanosized particles can also increase protein–protein interactions and aggregation, which can cause some health effects and change the material degradation mechanism. While proteins in some short-term studies (<6 h) even decrease material degradation due to shielding effects and better lubrication, they may increase degradation after longer time periods due to relatively slow binding, detachment, and combined corrosion processes. In-vitro material degradation studies of relatively corrosion-resistant alloys for biomedical applications should therefore include long-term studies, complexing agents or proteins, and realistic oxidative environments simulating inflammatory conditions.
Generation of therapeutic antisera for emerging viral infections
Authors: Schmidt Rebecca , Beltzig Lea C. , Sawatsky Bevan , Dolnik Olga , Dietzel Erik , Krähling Verena , Volz Asisa , Sutter Gerd , Becker Stephan , von Messling Veronika
Published: August 2018

The recent Ebola virus outbreak has highlighted the therapeutic potential of antisera and renewed interest in this treatment approach. While human convalescent sera may not be readily available in the early stages of an outbreak, antisera of animal origin can be produced in a short time frame. Here, we compared adjuvanted virus-like particles (VLP) with recombinant modified vaccinia virus Ankara and vesicular stomatitis virus (VSV), both expressing the Ebola virus antigens. The neutralizing antibody titers of rabbits immunized with adjuvanted VLPs were similar to those immunized with the replication-competent VSV, indicating that presentation of the antigen in its native conformation rather than de novo antigen expression is essential for production of functional antibodies. This approach also yielded high-titer antisera against Nipah virus glycoproteins, illustrating that it is transferable to other virus families. Multiple-step immunoglobulin G purification using a two-step 20–40% ammonium sulfate precipitation followed by protein A affinity chromatography resulted in 90% recovery of functionality and sustained in vivo stability. Adjuvanted VLP-based immunization strategies are thus a promising approach for the rapid generation of therapeutic antisera against emerging infections.
Sucrose Abstinence and Environmental Enrichment Effects on Mesocorticolimbic DARPP32 in Rats
Authors: Grimm Jeffrey W. , Glueck Edwin , Ginder Darren , Hyde Jeff , North Katherine , Jiganti Kyle
Published: August 2018

Dopamine- and cAMP-regulated neuronal phosphoprotein 32 kDa (DARPP32) is a signaling molecule that could serve as a molecular switch, promoting or restraining sucrose seeking. We measured DARPP32 and pThr34 DARPP32 in the brains of male Long-Evans rats with a history of sucrose self-administration followed by 1 or 30 days of abstinence and exposure to either overnight (acute) or one month (chronic) environmental enrichment (EE). Brains were extracted following a 1 h cue reactivity test or no exposure to the test environment. Micropunches (prelimbic, infralimbic, and anterior cingulate areas of the medial prefrontal cortex, orbitofrontal cortex, dorsal striatum, nucleus accumbens, and ventral tegmental area) were then processed using Western blot. Abstinence increased, while EE decreased, sucrose seeking. DARPP32 and pThr34 DARPP32 levels were affected by testing, abstinence, and/or EE in most regions. Especially salient results were observed in the nucleus accumbens core, a region associated with relapse behaviors. Both acute and chronic EE reduced DARPP32 in the nucleus accumbens core and acute EE increased the ratio of phosphorylated to total DARPP32. Degree of DARPP32 phosphorylation negatively correlated with sucrose seeking. These findings demonstrate a potential role for DARPP32 in mediating the “anti-craving” effect of EE.
Tyrosine kinase receptor TIE-1 mediates platinum resistance by promoting nucleotide excision repair in ovarian cancer
Authors: Ishibashi Masumi , Toyoshima Masafumi , Zhang Xuewei , Hasegawa-Minato Junko , Shigeta Shogo , Usui Toshinori , Kemp Christopher J. , Grandori Carla , Kitatani Kazuyuki , Yaegashi Nobuo
Published: August 2018

Platinum resistance is one of the most challenging problems in ovarian cancer treatment. High-throughput functional siRNA screening identified tyrosine kinase with immunoglobulin-like and EGF-like domains 1 (TIE-1) as a gene that confers cells resistant to cisplatin. Conversely enforced over-expression of TIE-1 was validated to decrease cisplatin sensitivity in multiple ovarian cancer cell lines and up-regulation of TIE-1 was correlated with poor prognosis and cisplatin resistance in patients with ovarian cancer. Mechanistically, TIE-1 up-regulates the nucleotide excision repair (NER) system mediated by xeroderma pigmentosum complementation group C (XPC), thereby leading to decreased susceptibility to cisplatin-induced cell death without affecting cisplatin uptake and excretion. Importantly potentiation of therapeutic efficacy by TIE-1 inhibition was selective to DNA-adduct-type chemotherapeutic platinum reagents. Therefore, TIE-1 is suggested to promote XPC-dependent NER, rendering ovarian cancer cells resistant to platinum. Accompanied with novel findings, TIE-1 could represent as a novel therapeutic target for platinum-resistant ovarian cancer.
Author Correction: Non-invasive multimodal optical coherence and photoacoustic tomography for human skin imaging
Authors: Chen Zhe , Rank Elisabet , Meiburger Kristen M. , Sinz Christoph , Hodul Andreas , Zhang Edward , Hoover Erich , Minneman Micheal , Ensher Jason , Beard Paul C. , Kittler Harald , Leitgeb Rainer A. , Drexler Wolfgang , Liu Mengyang
Published: August 2018

A correction to this article has been published and is linked from the HTML and PDF versions of this paper. The error has not been fixed in the paper.
Continuous infusion of PTH1–34 delayed fracture healing in mice
Authors: Yukata Kiminori , Kanchiku Tsukasa , Egawa Hiroshi , Nakamura Michihiro , Nishida Norihiro , Hashimoto Takahiro , Ogasa Hiroyoshi , Taguchi Toshihiko , Yasui Natsuo
Published: August 2018

Hyperparathyroidism, which is increased parathyroid hormone (PTH) levels in the blood, could cause delayed or non-union of bone fractures. But, no study has yet demonstrated the effects of excess continuous PTH exposure, such as that seen in hyperparathyroidism, for fracture healing. Continuous human PTH1–34 (teriparatide) infusion using an osmotic pump was performed for stabilized tibial fractures in eight-week-old male mice to determine the relative bone healing process compared with saline treatment. Radiographs and micro-computed tomography showed delayed but increased calcified callus formation in the continuous PTH1–34 infusion group compared with the controls. Histology and quantitative histomorphometry confirmed that continuous PTH1–34 treatment significantly increased the bone callus area at a later time point after fracture, since delayed endochondral ossification occurred. Gene expression analyses showed that PTH1–34 resulted in sustained Col2a1 and reduced Col10a1 expression, consistent with delayed maturation of the cartilage tissue during fracture healing. In contrast, continuous PTH1–34 infusion stimulated the expression of both Bglap and Acp5 through the healing process, in accordance with bone callus formation and remodeling. Mechanical testing showed that continuously administered PTH1–34 increased the maximum load on Day 21 compared with control mice. We concluded that continuous PTH1–34 infusion resulted in a delayed fracture healing process due to delayed callus cell maturation but ultimately increased biomechanical properties.
Chimeric yellow fever 17D-Zika virus (ChimeriVax-Zika) as a live-attenuated Zika virus vaccine
Authors: Giel-Moloney Maryann , Goncalvez Ana P. , Catalan John , Lecouturier Valerie , Girerd-Chambaz Yves , Diaz Fernando , Maldonado-Arocho Francisco , Gomila Raul C. , Bernard Marie-Clotilde , Oomen Ray , Delagrave Simon , Burdin Nicolas , Kleanthous Harold , Jackson Nicolas , Heinrichs Jon , Pugachev Konstantin V.
Published: August 2018

Zika virus (ZIKV) is an emerging mosquito-borne pathogen representing a global health concern. It has been linked to fetal microcephaly and other birth defects and neurological disorders in adults. Sanofi Pasteur has engaged in the development of an inactivated ZIKV vaccine, as well as a live chimeric vaccine candidate ChimeriVax-Zika (CYZ) that could become a preferred vaccine depending on future ZIKV epidemiology. This report focuses on the CYZ candidate that was constructed by replacing the pre-membrane and envelope (prM-E) genes in the genome of live attenuated yellow fever 17D vaccine virus (YF 17D) with those from ZIKV yielding a viable CYZ chimeric virus. The replication rate of CYZ in the Vero cell substrate was increased by using a hybrid YF 17D-ZIKV signal sequence for the prM protein. CYZ was highly attenuated both in mice and in human in vitro models (human neuroblastoma and neuronal progenitor cells), without the need for additional attenuating modifications. It exhibited significantly reduced viral loads in organs compared to a wild-type ZIKV and a complete lack of neuroinvasion following inoculation of immunodeficient A129 mice. A single dose of CYZ elicited high titers of ZIKV-specific neutralizing antibodies in both immunocompetent and A129 mice and protected animals from ZIKV challenge. The data indicate that CYZ is a promising vaccine candidate against ZIKV.
Structural basis for the complete resistance of the human prion protein mutant G127V to prion disease
Authors: Zheng Zhen , Zhang Meilan , Wang Yongheng , Ma Rongsheng , Guo Chenyun , Feng Liubin , Wu Jihui , Yao Hongwei , Lin Donghai
Published: August 2018

Prion diseases are caused by the propagation of misfolded cellular prion proteins (PrPs). A completely prion disease-resistant genotype, V127M129, has been identified in Papua New Guinea and verified in transgenic mice. To disclose the structural basis of the disease-resistant effect of the G127V mutant, we determined and compared the structural and dynamic features of the G127V-mutated human PrP (residues 91–231) and the wild-type PrP in solution. HuPrP(G127V) contains α1, α2 and α3 helices and a stretch-strand (SS) pattern comprising residues Tyr128-Gly131 (SS1) and Val161-Arg164 (SS2), with extending atomic distances between the SS1 and SS2 strands, and a structural rearrangement of the Tyr128 side chain due to steric hindrance of the larger hydrophobic side chain of Val127. The extended α1 helix gets closer to the α2 and α3 helices. NMR dynamics analysis revealed that Tyr128, Gly131 and Tyr163 underwent significant conformational exchanges. Molecular dynamics simulations suggest that HuPrP(G127V) prevents the formation of stable β-sheets and dimers. Unique structural and dynamic features potentially inhibit the conformational conversion of the G127V mutant. This work is beneficial for understanding the molecular mechanisms underlying the complete resistance of the G127V mutant to prion disease and for developing new therapeutics for prion disease.
Reduced fucosylation in the distal intestinal epithelium of mice subjected to chronic social defeat stress
Authors: Omata Yasuhiro , Aoki Reiji , Aoki-Yoshida Ayako , Hiemori Keiko , Toyoda Atsushi , Tateno Hiroaki , Suzuki Chise , Takayama Yoshiharu
Published: August 2018

Psychological stress can cause dysfunction of the gastrointestinal tract by regulating its interaction with central nervous system (brain-gut axis). Chronic social defeat stress (CSDS) is widely used to produce a rodent model of stress-induced human mood disorders and depression. We previously showed that CSDS significantly affects the intestinal ecosystem including cecal and fecal microbiota, intestinal gene expression profiles and cecal metabolite profiles. Here, we investigated whether the glycosylation pattern in the intestinal epithelium was affected in C57BL/6 mice exposed to CSDS (hereinafter referred to as CSDS mice). A lectin microarray analysis revealed that CSDS significantly reduced the reactivity of fucose-specific lectins (rAOL, TJA-II, rAAL, rGC2, AOL, AAL, rPAIIL and rRSIIL) with distal intestinal mucosa, but not with mucosa from proximal intestine and colon. Flow cytometric analysis confirmed the reduced TJA-II reactivity with intestinal epithelial cells in CSDS mice. In addition, distal intestine expression levels of the genes encoding fucosyltransferase 1 and 2 (Fut1 and Fut2) were downregulated in CSDS mice. These findings suggest that CSDS alters the fucosylation pattern in the distal intestinal epithelium, which could be used as a sensitive marker for CSDS exposure.
A novel glycosylated anti-CD20 monoclonal antibody from transgenic cattle
Authors: Zhang Ran , Tang Chenjun , Guo Huaizu , Tang Bo , Hou Sheng , Zhao Lei , Wang Jianwu , Ding Fangrong , Zhao Jianmin , Wang Haiping , Chen Zhongzhou , Dai Yunping , Li Ning
Published: August 2018

The monoclonal antibody (mAb) against CD20 known as Rituxan is widely used to treat autoimmune diseases and lymphomas. However, further application of Rituxan faces challenges of high production cost, which limits its availability in developing countries. Here, we report a new approach for large production of a recombinant anti-CD20 mAb in the milk of transgenic cattle (at a yield of up to ~6.8 mg/mL), with ~80% recovery rate and >99% purity. Crystallography study showed that our recombinant mAb is structurally nearly identical to Rituxan with only minor differences in N-linked glycosylation pattern. Functional study showed that, while our mAb shared similar target-cell binding capacities and complement-dependent cytotoxicity with Rituxan, our product exhibited a higher binding affinity for FcγRIIIα and a greater antibody-dependent cellular cytotoxicity. Accordingly, our recombinant mAb demonstrated a superior efficacy over Rituxan against B-cell lymphomas in severe combined immunodeficiency mice. Taken together, our data supports transgenic cattle as a novel model for cost-competitive, large-scale production of therapeutic antibodies.
Author Correction: Design and Computational Modeling of a Modular, Compliant Robotic Assembly for Human Lumbar Unit and Spinal Cord Assistance
Authors: Agarwal Gunjan , Robertson Matthew A. , Sonar Harshal , Paik Jamie
Published: August 2018

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Author Correction: Wound Healing in Streptozotocin-Induced Diabetic Rats Using Atmospheric-Pressure Argon Plasma Jet
Authors: Cheng Kuang-Yao , Lin Zhi-Hua , Cheng Yu-Pin , Chiu Hsien-Yi , Yeh Nai-Lun , Wu Tung-Kung , Wu Jong-Shinn
Published: August 2018

A correction to this article has been published and is linked from the HTML and PDF versions of this paper. The error has not been fixed in the paper.
Integration of gene interaction information into a reweighted random survival forest approach for accurate survival prediction and survival biomarker discovery
Authors: Wang Wei , Liu Wei
Published: August 2018

Accurately predicting patient risk and identifying survival biomarkers are two important tasks in survival analysis. For the emerging high-throughput gene expression data, random survival forest (RSF) is attracting more and more attention as it not only shows excellent performance on survival prediction problems with high-dimensional variables, but also is capable of identifying important variables according to variable importance automatically calculated within the algorithm. However, RSF still suffers from some problems such as limited predictive accuracy on independent datasets and limited biological interpretation of survival biomarkers. In this study, we integrated gene interaction information into a Reweighted RSF model (RRSF) to improve predictive accuracy and identify biologically meaningful survival markers. We applied RRSF to the prediction of patients with glioblastoma multiforme (GBM) and esophageal squamous cell carcinoma (ESCC). With a reconstructed global pathway network and an mRNA-lncRNA co-expression network as the prior gene interaction information, RRSF showed better overall predictive performance than RSF on three GBM and two ESCC datasets. In addition, RRSF identified a two-gene and three-lncRNA signature, which showed robust prognostic values and had high biological relevance to the development of GBM and ESCC, respectively.
Normal limits of home measured spatial gait parameters of the elderly population and their association with health variables
Authors: Herrero-Larrea Alexandra , Miñarro Antonio , Narvaiza Leire , Gálvez-Barrón César , León Natalia Gonzalo , Valldosera Esther , Felipe Elisabet , Valverde Rosa Ana , Kruse Liane , Sabater Joan Bosch , Rodríguez-Molinero Alejandro
Published: August 2018

Gait studies in the elderly population have been always conducted in gait labs or spacious clinical facilities, which influence gait parameters, and also implies that the participants have to be able to move to these facilities. Indoors gait characteristics of the elderly population have been very little studied. In this study, we aim to define the normal limits of the spatial gait parameters of the elderly, when walking at home, and to analyze relationship existing between the spatial gait parameters to other health variables. For such purpose, we conducted a transversal study on a probabilistic sample of 431 Spanish community-dwelling older, in which the spatial gait parameters were recorded by using an ink footprints method. We found that the mean stride length indoors was 88.47 cm (SD:26.05 cm; mean CI95%:85.52–91.41 cm), and the mean step width was 10.34 cm (SD:4.37 cm; mean CI95%:9.84–10.83 cm). Stride length was shorter in women and the oldest group, and was significantly influenced by the strength, balance, and physical activity. Stride width was larger in the oldest group and mainly affected by balance. A composite parameter including width and normalized stride length was independent from sex, and strongly differentiated between age groups. This parameter was affected by strength.
Some things never change: multi-decadal stability in humpback whale calling repertoire on Southeast Alaskan foraging grounds
Authors: Fournet Michelle E. H. , Gabriele Christine M. , Culp David C. , Sharpe Fred , Mellinger David K. , Klinck Holger
Published: August 2018

Investigating long term trends in acoustic communication is essential for understanding the role of sound in social species. Humpback whales are an acoustically plastic species known for producing rapidly-evolving song and a suite of non-song vocalizations (“calls”) containing some call types that exhibit short-term stability. By comparing the earliest known acoustic recordings of humpback whales in Southeast Alaska (from the 1970’s) with recordings collected in the 1990’s, 2000’s, and 2010’s, we investigated the long-term repertoire stability of calls on Southeast Alaskan foraging grounds. Of the sixteen previously described humpback whale call types produced in Southeast Alaska, twelve were detected in both 1976 and 2012, indicating stability over a 36-year time period; eight call types were present in all four decades and every call type was present in at least three decades. We conclude that the conservation of call types at this temporal scale is indicative of multi-generational persistence and confirms that acoustic communication in humpback whales is comprised of some highly stable call elements in strong contrast to ever-changing song.
Hybridization and extensive mitochondrial introgression among fire salamanders in peninsular Italy
Authors: Bisconti Roberta , Porretta Daniele , Arduino Paola , Nascetti Giuseppe , Canestrelli Daniele
Published: August 2018

Discordance between mitochondrial and nuclear patterns of population genetic structure is providing key insights into the eco-evolutionary dynamics between and within species, and their assessment is highly relevant to biodiversity monitoring practices based on DNA barcoding approaches. Here, we investigate the population genetic structure of the fire salamander Salamandra salamandra in peninsular Italy. Both mitochondrial and nuclear markers clearly identified two main population groups. However, nuclear and mitochondrial zones of geographic transition between groups were located 600 km from one another. Recent population declines in central Italy partially erased the genetic imprints of past hybridization dynamics. However, the overall pattern of genetic variation, together with morphological and fossil data, suggest that a rampant mitochondrial introgression triggered the observed mitonuclear discordance, following a post-glacial secondary contact between lineages. Our results clearly show the major role played by reticulate evolution in shaping the structure of Salamandra salamandra populations and, together with similar findings in other regions of the species’ range, contribute to identify the fire salamander as a particularly intriguing case to investigate the complexity of mechanisms triggering patterns of mitonuclear discordance in animals.
Limiting asymmetric hearing improves benefits of bilateral hearing in children using cochlear implants
Authors: Polonenko Melissa Jane , Papsin Blake Croll , Gordon Karen Ann
Published: August 2018

Neurodevelopmental changes occur with asymmetric hearing loss, limiting binaural/spatial hearing and putting children at risk for social and educational challenges. These deficits may be mitigated by providing bilateral hearing in children through auditory prostheses. Effects on speech perception and spatial hearing were measured in a large cohort of >450 children who were deaf and used bilateral cochlear implants or bimodal devices (one cochlear implant and a contralateral hearing aid). Results revealed an advantage of bilateral over unilateral device use but this advantage decreased as hearing in the two ears became increasingly asymmetric. Delayed implantation of an ear with severe to profound deafness allowed asymmetric hearing, creating aural preference for the better hearing ear. These findings indicate that bilateral input with the most appropriate device for each ear should be provided early and without delay during development.
Microvascular dysfunction in ankylosing spondylitis is associated with disease activity and is improved by anti-TNF treatment
Authors: Batko Bogdan , Maga Pawel , Urbanski Karol , Ryszawa-Mrozek Natalia , Schramm-Luc Agata , Koziej Mateusz , Mikolajczyk Tomasz , McGinnigle Eilidh , Czesnikiewicz-Guzik Marta , Ceranowicz Piotr , Guzik Tomasz J.
Published: August 2018

Ankylosing spondylitis (AS) is associated with high cardiovascular morbidity and mortality. Recent studies indicate that microvascular dysfunction may underlie cardiovascular risk in AS. We hypothesized, that microvascular morphology and dysfunction is linked to AS activity and is modifiable by TNF-α inhibitor (TNFi) treatment. Functional Laser Doppler Flowmetry with post-occlusive reactive hyperemia, and structural nailfold capillaroscopy were performed in 54 patients with AS and 28 matched controls. Active AS was diagnosed based on BASDAI ≥ 4 (n = 37). Effects of 3-month TNFi on microcirculation in active AS were studied. AS was associated with prolonged time to peak hyperemia compared to healthy controls. High disease activity was associated with increased time to peak hyperemia and decreased peak hyperemia when compared to patients with inactive AS. In capillaroscopy, AS was associated with morphological abnormalities indicating increased neoangiogenesis and pericapillary edema compared to controls. Microvascular function improved following 3 months of TNFi in reference to basal flow as well as post-occlusive parameters. TNFi reduced pericapillary edema, while other parameters of capillary morphology remained unchanged. Microvascular dysfunction and capillary neovascular formation are associated with disease activity of AS. Anti-TNF-α treatment may restore microcirculation function and capillary edema but does not modify microvascular structural parameters.
Rational metabolic-flow switching for the production of exogenous secondary metabolites in bamboo suspension cells
Authors: Nomura Taiji , Ogita Shinjiro , Kato Yasuo
Published: August 2018

The synthetic biology-driven production of high-value plant secondary metabolites in microbial hosts has attracted extensive attention despite various challenges, including correct protein expression and limited supplies of starting materials. In contrast, plant cell cultures are rarely used for this purpose owing to their slow proliferation rates and laborious transformation processes. Here, we propose a “rational metabolic-flow switching” strategy to efficiently produce exogenous secondary metabolites using suspension-cultured bamboo (Phyllostachys nigra; Pn) cells as model production hosts. The Pn cells biosynthesise hydroxycinnamic acid amides (HCAAs) of putrescine as major secondary metabolites, which indicates that the phenylpropanoid and polyamine biosynthetic pathways are highly active and that the Pn cells may produce alternative secondary metabolites derived from those pathways. Stable transformants of Pn cells expressing agmatine coumaroyltransferase of barley (Hordeum vulgare) were generated with the expectation of metabolic-flow switching from HCAAs of putrescine to those of agmatine. In the recombinant Pn cells, the levels of HCAAs of putrescine decreased and the HCAAs of agmatine were produced instead. The production titre of the major product, p-coumaroylagmatine, reached approximately 360 mg/L, providing a proof-of-concept for the usefulness of “rational metabolic-flow switching” in synthetic biology using plant cell hosts.
Evolution shapes the responsiveness of the D-box enhancer element to light and reactive oxygen species in vertebrates
Authors: Pagano Cristina , Siauciunaite Rima , Idda Maria L. , Ruggiero Gennaro , Ceinos Rosa M. , Pagano Martina , Frigato Elena , Bertolucci Cristiano , Foulkes Nicholas S. , Vallone Daniela
Published: August 2018

The circadian clock is a highly conserved cell-autonomous mechanism that directs daily rhythms in most aspects of biology. Daily entrainment by environmental signals, notably light, is essential for its function. However, our understanding of the mechanisms and the evolution of photic entrainment remains incomplete. Fish represent attractive models for exploring how light regulates the circadian clock due to the direct light sensitivity of their peripheral clocks. Central to this property is the light induced expression of clock genes that is mediated by D-box enhancer elements. Here, using zebrafish cells, we reveal that the light responsive D-box enhancer serves as a nuclear target for reactive oxygen species (ROS). We demonstrate that exposure to short wavelengths of visible light triggers increases in ROS levels via NADPH oxidase activity. Elevated ROS activates the JNK and p38 MAP kinases and in turn, induces clock gene expression via the D-box. In blind cavefish and mammals, where peripheral clocks are no longer entrained by direct illumination, ROS levels are still increased upon light exposure. However, in these species ROS no longer induces D-box driven clock gene transcription. Thus, during evolution, alterations in ROS-responsive signal transduction pathways underlie fundamental changes in peripheral clock photoentrainment.
Single-neuronal cell culture and monitoring platform using a fully transparent microfluidic DEP device
Authors: Kim Hyungsoo , Lee In-Kyu , Taylor Kendra , Richters Karl , Baek Dong-Hyun , Ryu Jae Ha , Cho Sang June , Jung Yei Hwan , Park Dong-Wook , Novello Joseph , Bong Jihye , Suminski Aaron J. , Dingle Aaron M. , Blick Robert H. , Williams Justin C. , Dent Erik W. , Ma Zhenqiang
Published: August 2018

Dielectrophoresis using multi-electrode arrays allows a non-invasive interface with biological cells for long-term monitoring of electrophysiological parameters as well as a label-free and non-destructive technique for neuronal cell manipulation. However, experiments for neuronal cell manipulation utilizing dielectrophoresis have been constrained because dielectrophoresis devices generally function outside of the controlled environment (i.e. incubator) during the cell manipulation process, which is problematic because neurons are highly susceptible to the properties of the physiochemical environment. Furthermore, the conventional multi-electrode arrays designed to generate dielectrophoretic force are often fabricated with non-transparent materials that confound live-cell imaging. Here we present an advanced single-neuronal cell culture and monitoring platform using a fully transparent microfluidic dielectrophoresis device for the unabated monitoring of neuronal cell development and function. The device is mounted inside a sealed incubation chamber to ensure improved homeostatic conditions and reduced contamination risk. Consequently, we successfully trap and culture single neurons on a desired location and monitor their growth process over a week. The proposed single-neuronal cell culture and monitoring platform not only has significant potential to realize an in vitro ordered neuronal network, but also offers a useful tool for a wide range of neurological research and electrophysiological studies of neuronal networks.