bluuebunny/crossref_metadata_2025 huggingface dataset:
Outcomes of Combined Medical and Behavioral Treatments for Constipation Within a Specialty Outpatient Clinic
Authors: Hankinson Jessica C. , Borden Lindsay , Allen Taryn , Santo Domingo Lisa , Oliva-Hemker Maria , Mathews Tara , Brock Kristen , Perry-Parrish Carisa , Reynolds Elizabeth K. , Specht Matthew , Ostrander Rick
Published: August 2017

Chronic constipation is a common and challenging childhood problem. Effective treatment for chronic constipation and fecal incontinence includes a combination of medical and behavioral treatments. This study evaluates the outcomes associated with providing these treatments in a specialty outpatient clinic consisting of a nurse practitioner and behavioral psychologist. Fifty-seven patients were followed up for initial and follow-up appointments in this multidisciplinary chronic constipation clinic. In addition, 65% of the patients did not return after the initial visit; from this cohort, 21 patients (20%) were randomly selected and outcomes were derived by phone interview. Results showed a reduction in bowel accident frequency, abdominal pain, use of diapers, and perceptions of having constipation. In most cases, these outcomes were achieved after a single visit; however, a more extended treatment was required to achieve optimal benefits for patients who initially presented with frequent bowel accidents. These results suggest that a multidisciplinary clinic comprised of a nurse practitioner and behavioral psychologist provides an effective and efficient way to provide a combined treatment protocol for constipation in children. Future research should evaluate long-term effects of this intervention on constipation symptoms and overall health care utilization.
Erratum: iPS-derived MSCs from an expandable bank to deliver a prodrug-converting enzyme that limits growth and metastases of human breast cancers
Authors: Ullah M , Kuroda Y , Bartosh T J , Liu F , Zhao Q , Gregory C , Reger R , Xu J , Lee R H , Prockop D J
Published: August 2017

Correction to: Cell Death Discovery (2017) 3, 16064; doi:10.1038/cddiscovery.2016.64; published online 6 February 2017 Since the publication of this article, it has been noted that the following acknowledgement was missing: Mujib Ullah and Yasumasa Kuroda contributed equally to the work. The authorsapologise for this error.
Amyloid single-cell cytotoxicity assays by nanomotion detection
Authors: Ruggeri Francesco S , Mahul-Mellier Anne-Laure , Kasas Sandor , Lashuel Hilal A , Longo Giovanni , Dietler Giovanni
Published: August 2017

Cells are extremely complex systems able to actively modify their metabolism and behavior in response to environmental conditions and stimuli such as pathogenic agents or drugs. The comprehension of these responses is central to understand the molecular bases of human pathologies, including amyloid misfolding diseases. Conventional bulk biological assays are limited by intrinsic cellular heterogeneity in gene, protein and metabolite expression, and can investigate only indirectly cellular reactions in non-physiological conditions. Here we employ a label-free nanomotion sensor to study single neuroblastoma cells exposed to extracellular monomeric and amyloid α-synuclein species in real-time and in physiological conditions. Combining this technique with fluorescence microscopy, we demonstrate multispecies cooperative cytotoxic effect of amyloids and aggregate-induced loss of cellular membrane integrity. Notably, the method can study cellular reactions and cytotoxicity an order of magnitude faster, and using 100-fold smaller volume of reagents when compared to conventional bulk analyses. This rapidity and sensitivity will allow testing novel pharmacological approaches to stop or delay a wide range of human diseases.
MicroRNA-34a/EGFR axis plays pivotal roles in lung tumorigenesis
Authors: Li Y-L , Liu X-M , Zhang C-Y , Zhou J-B , Shao Y , Liang C , Wang H-M , Hua Z-Y , Lu S-D , Ma Z-L
Published: August 2017

MicroRNAs (miRNAs) are vital in the regulation of tumor progression and invasion. Dysregulation of miRNAs has been linked to the development of various types of human cancers, including non-small-cell lung cancer (NSCLC). However, the effect of miRNA-34a (miR-34a), a key regulator of tumor suppression, on the tumorigenesis of NSCLC has not been fully elaborated. Herein, we reveal that miR-34a is significantly downregulated in NSCLC tissues and cell lines, suggesting that miR-34a might function as a tumor suppressor in lung cancer. We also confirmed that epidermal growth factor receptor (EGFR) is a direct target of miR-34a, and our data reveal that siRNA knockdown of EGFR can inhibit cell proliferation, promote apoptosis and arrest cell-cycle progression. In addition, EGFR can reverse the suppressive function of miR-34a overexpression on proliferation and cell apoptosis. Furthermore, in vivo experiments demonstrated that miR-34a suppress tumor growth, both in the A549 xenograft model, as well as in the metastatic tumors in nude mice. Taken together, our findings suggest that miR-34a inhibits NSCLC tumor growth and metastasis through targeting EGFR.
A probabilistic assessment of the rapidity of PETM onset
Authors: Kirtland Turner Sandra , Hull Pincelli M. , Kump Lee R. , Ridgwell Andy
Published: August 2017

Knowledge of the onset duration of the Paleocene-Eocene Thermal Maximum—the largest known greenhouse-gas-driven global warming event of the Cenozoic—is central to drawing inferences for future climate change. Single-foraminifera measurements of the associated carbon isotope excursion from Maud Rise (South Atlantic Ocean) are controversial, as they seem to indicate geologically instantaneous carbon release and anomalously long ocean mixing. Here, we fundamentally reinterpret this record and extract the likely PETM onset duration. First, we employ an Earth system model to illustrate how the response of ocean circulation to warming does not support the interpretation of instantaneous carbon release. Instead, we use a novel sediment-mixing model to show how changes in the relative population sizes of calcareous plankton, combined with sediment mixing, can explain the observations. Furthermore, for any plausible PETM onset duration and sampling methodology, we place a probability on not sampling an intermediate, syn-excursion isotopic value. Assuming mixed-layer carbonate production continued at Maud Rise, we deduce the PETM onset was likely <5 kyr.
Ultra-large single crystals by abnormal grain growth
Authors: Kusama Tomoe , Omori Toshihiro , Saito Takashi , Kise Sumio , Tanaka Toyonobu , Araki Yoshikazu , Kainuma Ryosuke
Published: August 2017

Producing a single crystal is expensive because of low mass productivity. Therefore, many metallic materials are being used in polycrystalline form, even though material properties are superior in a single crystal. Here we show that an extraordinarily large Cu-Al-Mn single crystal can be obtained by abnormal grain growth (AGG) induced by simple heat treatment with high mass productivity. In AGG, the sub-boundary energy introduced by cyclic heat treatment (CHT) is dominant in the driving pressure, and the grain boundary migration rate is accelerated by repeating the low-temperature CHT due to the increase of the sub-boundary energy. With such treatment, fabrication of single crystal bars 70 cm in length is achieved. This result ensures that the range of applications of shape memory alloys will spread beyond small-sized devices to large-scale components and may enable new applications of single crystals in other metallic and ceramics materials having similar microstructural features.
Conformal metasurface-coated dielectric waveguides for highly confined broadband optical activity with simultaneous low-visibility and reduced crosstalk
Authors: Jiang Zhi Hao , Kang Lei , Werner Douglas H.
Published: August 2017

The ability to achieve simultaneous control over the various electromagnetic properties of dielectric waveguides, including mode confinement, polarization, scattering signature, and crosstalk, which are critical to system miniaturization, diversity in functionality, and non-invasive integration, has been a highly sought after yet elusive goal. Currently existing methods, which rely on three-dimensional artificial cores or claddings and/or structural chirality, provide efficient paths for obtaining either highly confined modes, optical activity, or a low-scattering signature, but at the expense of increased propagation loss, form factor and weight. Here, by tailoring the unique anisotropy and exploiting the inter-cell coupling of metasurface coatings, we report a unified approach for simultaneously controlling the diverse optical properties of dielectric waveguides. The experimentally demonstrated highly confined sub-wavelength dielectric waveguide with a low-visibility and broadband optical activity represents a transformative wave manipulation capability with far reaching implications, offering new pathways for future miniaturization of dielectric waveguide-based systems with simultaneous polarization and scattering control.
Tumor-selective catalytic nanomedicine by nanocatalyst delivery
Authors: Huo Minfeng , Wang Liying , Chen Yu , Shi Jianlin
Published: August 2017

Tumor cells metabolize in distinct pathways compared with most normal tissue cells. The resulting tumor microenvironment would provide characteristic physiochemical conditions for selective tumor modalities. Here we introduce a concept of sequential catalytic nanomedicine for efficient tumor therapy by designing and delivering biocompatible nanocatalysts into tumor sites. Natural glucose oxidase (GOD, enzyme catalyst) and ultrasmall Fe3O4 nanoparticles (inorganic nanozyme, Fenton reaction catalyst) have been integrated into the large pore-sized and biodegradable dendritic silica nanoparticles to fabricate the sequential nanocatalyst. GOD in sequential nanocatalyst could effectively deplete glucose in tumor cells, and meanwhile produce a considerable amount of H2O2 for subsequent Fenton-like reaction catalyzed by Fe3O4 nanoparticles in response to mild acidic tumor microenvironment. Highly toxic hydroxyl radicals are generated through these sequential catalytic reactions to trigger the apoptosis and death of tumor cells. The current work manifests a proof of concept of catalytic nanomedicine by approaching selectivity and efficiency concurrently for tumor therapeutics.
HSP70-Hrd1 axis precludes the oncorepressor potential of N-terminal misfolded Blimp-1s in lymphoma cells
Authors: Wang Wen-Fang , Yan Li , Liu Zhao , Liu Lan-Xuan , Lin Jian , Liu Zhi-Yin , Chen Xiong-Ping , Zhang Wu , Xu Zi-Zhen , Shi Ting , Li Jun-Min , Zhao Yi-Lei , Meng Guoyu , Xia Yi , Li Jian-Yong , Zhu Jiang
Published: August 2017

B lymphocyte-induced maturation protein-1 (Blimp-1) ensures B-cell differentiation into the plasma cell stage, and its instability constitutes a crucial oncogenic element in certain aggressive cases of activated B cell-like diffuse large B-cell lymphoma (ABC-DLBCL). However, the underlying degradation mechanisms and their possible therapeutic relevance remain unexplored. Here, we show that N-terminal misfolding mutations in ABC-DLBCL render Blimp-1 protein susceptible to proteasome-mediated degradation but spare its transcription-regulating activity. Mechanistically, whereas wild-type Blimp-1 metabolism is triggered in the nucleus through PML-mediated sumoylation, the degradation of lymphoma-associated mutants is accelerated by subversion of this pathway to Hrd1-mediated cytoplasmic sequestration and ubiquitination. Screening experiments identifies the heat shock protein 70 (HSP70) that selects Blimp-1 mutants for Hrd1 association, and HSP70 inhibition restores their nuclear accumulation and oncorepressor activities without disrupting normal B-cell maturation. Therefore, HSP70-Hrd1 axis represents a potential therapeutic target for restoring the oncorepressor activity of unstable lymphoma-associated Blimp-1 mutants.
Activated Mesenchymal Stem Cells Interact with Antibiotics and Host Innate Immune Responses to Control Chronic Bacterial Infections
Authors: Johnson Valerie , Webb Tracy , Norman Annalis , Coy Jonathan , Kurihara Jade , Regan Daniel , Dow Steven
Published: August 2017

Chronic bacterial infections associated with biofilm formation are often difficult to resolve without extended courses of antibiotic therapy. Mesenchymal stem cells (MSC) exert antibacterial activity in vitro and in acute bacterial infection models, but their activity in chronic infection with biofilm models has not been previously investigated. Therefore, we studied the effects of MSC administration in mouse and dog models of chronic infections associated with biofilms. Mice with chronic Staphylococcus aureus implant infections were treated by i.v. administration of activated or non-activated MSC, with or without antibiotic therapy. The most effective treatment protocol was identified as activated MSC co-administered with antibiotic therapy. Activated MSC were found to accumulate in the wound margins several days after i.v. administration. Macrophages in infected tissues assumed an M2 phenotype, compared to untreated infections which contained predominately M1 macrophages. Bacterial killing by MSC was found to be mediated in part by secretion of cathelicidin and was significantly increased by antibiotics. Studies in pet dogs with spontaneous chronic multi drug-resistant wound infections demonstrated clearance of bacteria and wound healing following repeated i.v. administration of activated allogeneic canine MSC. Thus, systemic therapy with activated MSC may be an effective new, non-antimicrobial approach to treatment of chronic, drug-resistant infections.
Changing Epidemic of HIV and Syphilis Among Resident and Migrant Men Who Have Sex with Men in Jiangsu, China
Authors: Chen Yuheng , Tang Weiming , Chen Lusi , Shi Lingen , Liu Xiaoyan , Xu Jinshui , Hu Haiyang , Yang Haitao , Huan Xiping , Fu Gengfeng
Published: August 2017

Men who have sex with men (MSM) in China face high rates of HIV and syphilis infection exacerbated by internal migration. Studies on the differences of HIV and syphilis epidemics changing trends in high-risk behaviors and geographic distribution between resident and migrant MSM in Jiangsu, China were conducted. MSM were recruited from 14 surveillance sites in the serial cross-sectional study. Data on demographics, sexual behaviors, HIV and syphilis prevalence were collected. Participants were classified as residents or migrants based on household registration. During 2010–2014, 19,750 MSM were investigated. Engaged in anal sex (76.3% to 80.2%, P < 0.01) as well as received HIV-related services (72.1% to 79.2%, P < 0.01) were increasing. In contrast, engaged in commercial anal sex with males (7.4% to 5.0%, P < 0.01) and drug use (1.6% to 0.8%, P < 0.01) were decreasing. HIV prevalence ranged between 8.6% to 9.6%, while syphilis prevalence decreased over time (13.4–6.8%, P < 0.01). Further, we found that migrant MSM were more likely to engage in condomless anal sex, also had a higher HIV and syphilis prevalence than resident. During the study period, while syphilis prevalence decreased, higher rates of risk behaviors among migrant MSM called for targeted intervention strategies to reduce the HIV transmission.
Tau plasma levels in subjective cognitive decline: Results from the DELCODE study
Authors: Müller Stephan , Preische Oliver , Göpfert Jens C. , Yañez Viviana A. Carcamo , Joos Thomas O. , Boecker Henning , Düzel Emrah , Falkai Peter , Priller Josef , Buerger Katharina , Catak Cihan , Janowitz Daniel , Heneka Michael T. , Brosseron Frederic , Nestor Peter , Peters Oliver , Menne Felix , Schipke Carola G. , Schneider Anja , Spottke Annika , Fließbach Klaus , Kilimann Ingo , Teipel Stefan , Wagner Michael , Wiltfang Jens , Jessen Frank , Laske Christoph
Published: August 2017

Previous studies have demonstrated increased tau plasma levels in patients with Alzheimer’s disease (AD) and mild cognitive impairment (MCI) due to AD. Much less is known whether increased tau plasma levels can already be detected in the pre-MCI stage of subjective cognitive decline (SCD). In the present study we measured tau plasma levels in 111 SCD patients and 134 age- and gender-matched cognitively healthy controls participating in the DZNE (German Center for Neurodegenerative Diseases) longitudinal study on cognition and dementia (DELCODE). Tau plasma levels were measured using ultra-sensitive, single-molecule array (Simoa) technology. We found no significant different tau plasma levels in SCD (3.4 pg/ml) compared with healthy controls (3.6 pg/ml) after controlling for age, gender, and education (p = 0.137). In addition, tau plasma levels did not correlate with Aβ42 (r = 0.073; p = 0.634), tau (r = −0.179; p = 0.240), and p-tau181 (r = −0.208; p = 0.171) cerebrospinal fluid (CSF) levels in a subgroup of 45 SCD patients with available CSF. In conclusion, plasma tau is not increased in SCD patients. In addition, the lack of correlation between tau in plasma and CSF in the examined cohort suggests that tau levels are affected by different factors in both biofluids.
Effects of alcohol drinking and smoking on pancreatic ductal adenocarcinoma mortality: A retrospective cohort study consisting of 1783 patients
Authors: Zhang Shuisheng , Wang Chengfeng , Huang Huang , Jiang Qinglong , Zhao Dongbing , Tian Yantao , Ma Jie , Yuan Wei , Sun Yuemin , Che Xu , Zhang Jianwei , Chen Haibo , Zhao Yajie , Chu Yunmian , Zhang Yawei , Chen Yingtai
Published: August 2017

The effects of alcohol drinking and smoking on pancreatic ductal adenocarcinoma (PDAC) mortality are contradictory. Individuals who were diagnosed as PDAC and hospitalized at the China National Cancer Center between January 1999 and January 2016 were identified and included in the study. Ultimately, 1783 consecutive patients were included in the study. Patients were categorized as never, ex-drinkers/smokers or current drinkers/smokers. Hazard ratios (HRs) of all-cause mortality and 95% confidence intervals (CIs) were estimated using Cox proportional hazards models. Compared with never drinkers, the HRs were 1.25 for ever drinkers, 1.24 for current drinkers, and 1.33 for ex-drinkers (trend P = 0.031). Heavy drinking and smoking period of 30 or more years were positive prognostic factors for PDAC. For different smoking and alcohol drinking status, only subjects who are both current smokers and current drinkers (HR, 1.45; 95% CI, 1.03–2.05) were associated with reduced survival after PDAC compared to those who were never smokers and never drinkers. Patients who are alcohol drinkers and long-term smokers before diagnosis have a significantly higher risk of PDAC mortality. Compared to those who neither smoker nor drink, only patients who both smokers and drinkers were associated with reduced survival from PDAC.
Detecting Photosymbiosis in Fossil Scleractinian Corals
Authors: Tornabene Chiara , Martindale Rowan C. , Wang Xingchen T. , Schaller Morgan F.
Published: August 2017

The evolutionary success of reef-building corals is often attributed to photosymbiosis, a mutualistic relationship scleractinian corals developed with zooxanthellae; however, because zooxanthellae are not fossilized, it is difficult (and contentious) to determine whether ancient corals harbored symbionts. In this study, we analyze the δ15N of skeletal organic matrix in a suite of modern and fossil scleractinian corals (zooxanthellate- and azooxanthellate-like) with varying levels of diagenetic alteration. Significantly, we report the first analyses that distinguish shallow-water zooxanthellate and deep-water azooxanthellate fossil corals. Early Miocene (18–20 Ma) corals exhibit the same nitrogen isotopic ratio offset identified in modern corals. These results suggest that the coral organic matrix δ15N proxy can successfully be used to detect photosymbiosis in the fossil record. This proxy will significantly improve our ability to effectively define the evolutionary relationship between photosymbiosis and reef-building through space and time. For example, Late Triassic corals have symbiotic values, which tie photosymbiosis to major coral reef expansion. Furthermore, the early Miocene corals from Indonesia have low δ15N values relative to modern corals, implying that the west Pacific was a nutrient-depleted environment and that oligotrophy may have facilitated the diversification of the reef builders in the Coral Triangle.
Methods for estimating disease transmission rates: Evaluating the precision of Poisson regression and two novel methods
Authors: Kirkeby Carsten , Halasa Tariq , Gussmann Maya , Toft Nils , Græsbøll Kaare
Published: August 2017

Precise estimates of disease transmission rates are critical for epidemiological simulation models. Most often these rates must be estimated from longitudinal field data, which are costly and time-consuming to conduct. Consequently, measures to reduce cost like increased sampling intervals or subsampling of the population are implemented. To assess the impact of such measures we implement two different SIS models to simulate disease transmission: A simple closed population model and a realistic dairy herd including population dynamics. We analyze the accuracy of different methods for estimating the transmission rate. We use data from the two simulation models and vary the sampling intervals and the size of the population sampled. We devise two new methods to determine transmission rate, and compare these to the frequently used Poisson regression method in both epidemic and endemic situations. For most tested scenarios these new methods perform similar or better than Poisson regression, especially in the case of long sampling intervals. We conclude that transmission rate estimates are easily biased, which is important to take into account when using these rates in simulation models.
The Value of Coastal Wetlands for Flood Damage Reduction in the Northeastern USA
Authors: Narayan Siddharth , Beck Michael W. , Wilson Paul , Thomas Christopher J. , Guerrero Alexandra , Shepard Christine C. , Reguero Borja G. , Franco Guillermo , Ingram Jane Carter , Trespalacios Dania
Published: August 2017

As exposure to coastal hazards increases there is growing interest in nature-based solutions for risk reduction. This study uses high-resolution flood and loss models to quantify the impacts of coastal wetlands in the northeastern USA on (i) regional flood damages by Hurricane Sandy and (ii) local annual flood losses in Barnegat Bay in Ocean County, New Jersey. Using an extensive database of property exposure, the regional study shows that wetlands avoided $625 Million in direct flood damages during Hurricane Sandy. The local study combines these models with a database of synthetic storms in Ocean County and estimates a 16% average reduction in annual flood losses by salt marshes with higher reductions at lower elevations. Together, the studies quantify the risk reduction ecosystem services of marsh wetlands. Measuring these benefits in collaboration with the risk modelling industry is crucial for assessing risk accurately and, where appropriate, aligning conservation and risk reduction goals.
Responses of renal hemodynamics and tubular functions to acute sodium–glucose cotransporter 2 inhibitor administration in non-diabetic anesthetized rats
Authors: Ansary Tuba M. , Fujisawa Yoshihide , Rahman Asadur , Nakano Daisuke , Hitomi Hirofumi , Kobara Hideki , Masaki Tsutomu , Titze Jens M. , Kitada Kento , Nishiyama Akira
Published: August 2017

The aim of this study is to examine the effects of acute administration of luseogliflozin, the sodium–glucose cotransporter 2 (SGLT2) inhibitor, on renal hemodynamics and tubular functions in anesthetized non-diabetic Sprague Dawley (SD) rats and 5/6 nephrectomized (Nx) SD rats. Renal blood flow (RBF), mean arterial pressure (MAP), and heart rate (HR) were continuously measured and urine was collected directly from the left ureter. Intraperitoneal injection of luseogliflozin (0.9 mg kg−1) did not change MAP, HR, RBF, or creatinine clearance (CrCl) in SD rats (n = 7). Luseogliflozin significantly increased urine volume, which was associated with significantly increased urinary glucose excretion rates (P < 0.001). Similarly, luseogliflozin significantly increased urinary sodium excretion (from 0.07 ± 0.01 µmol min−1 at baseline to 0.76 ± 0.08 µmol min−1 at 120 min; P < 0.001). Furthermore, luseogliflozin resulted in significantly increased urinary pH (P < 0.001) and decreased urinary osmolality and urea concentration (P < 0.001) in SD rats. Similarly, in Nx SD rats (n = 5–6), luseogliflozin significantly increased urine volume and urinary glucose excretion (P < 0.001) without altering MAP, HR, RBF, or CrCl. Luseogliflozin did not elicit any significant effects on the other urinary parameters in Nx SD rats. These data indicate that SGLT2 inhibitor elicits direct tubular effects in non-diabetic rats with normal renal functions.
Probing charge transfer between molecular semiconductors and graphene
Authors: Matković Aleksandar , Kratzer Markus , Kaufmann Benjamin , Vujin Jasna , Gajić Radoš , Teichert Christian
Published: August 2017

The unique density of states and exceptionally low electrical noise allow graphene-based field effect devices to be utilized as extremely sensitive potentiometers for probing charge transfer with adsorbed species. On the other hand, molecular level alignment at the interface with electrodes can strongly influence the performance of organic-based devices. For this reason, interfacial band engineering is crucial for potential applications of graphene/organic semiconductor heterostructures. Here, we demonstrate charge transfer between graphene and two molecular semiconductors, parahexaphenyl and buckminsterfullerene C60. Through in-situ measurements, we directly probe the charge transfer as the interfacial dipoles are formed. It is found that the adsorbed molecules do not affect electron scattering rates in graphene, indicating that charge transfer is the main mechanism governing the level alignment. From the amount of transferred charge and the molecular coverage of the grown films, the amount of charge transferred per adsorbed molecule is estimated, indicating very weak interaction.
Generation of a new infectious recombinant prion: a model to understand Gerstmann–Sträussler–Scheinker syndrome
Authors: Elezgarai Saioa R. , Fernández-Borges Natalia , Eraña Hasier , Sevillano Alejandro M. , Charco Jorge M. , Harrathi Chafik , Saá Paula , Gil David , Kong Qingzhong , Requena Jesús R. , Andréoletti Olivier , Castilla Joaquín
Published: August 2017

Human transmissible spongiform encephalopathies (TSEs) or prion diseases are a group of fatal neurodegenerative disorders that include Kuru, Creutzfeldt-Jakob disease, Gerstmann-Sträussler-Scheinker syndrome (GSS), and fatal familial insomnia. GSS is a genetically determined TSE caused by a range of mutations within the prion protein (PrP) gene. Several animal models, based on the expression of PrPs carrying mutations analogous to human heritable prion diseases, support that mutations might predispose PrP to spontaneously misfold. An adapted Protein Misfolding Cyclic Amplification methodology based on the use of human recombinant PrP (recPMCA) generated different self-propagating misfolded proteins spontaneously. These were characterized biochemically and structurally, and the one partially sharing some of the GSS PrPSc molecular features was inoculated into different animal models showing high infectivity. This constitutes an infectious recombinant prion which could be an invaluable model for understanding GSS. Moreover, this study proves the possibility to generate recombinant versions of other human prion diseases that could provide a further understanding on the molecular features of these devastating disorders.
Homogeneity evaluation of mesenchymal stem cells based on electrotaxis analysis
Authors: Kim Min Sung , Lee Mi Hee , Kwon Byeong-Ju , Kim Dohyun , Koo Min-Ah , Seon Gyeung Mi , Park Jong-Chul
Published: August 2017

Stem cell therapy that can restore function to damaged tissue, avoid host rejection and reduce inflammation throughout body without use of immunosuppressive drugs. The established methods were used to identify and to isolate specific stem cell markers by FACS or by immunomagnetic cell separation. The procedures for distinguishing population of stem cells took a time and needed many preparations. Here we suggest an electrotaxis analysis as a new method to evaluate the homogeneity of mesenchymal stem cells which can observe the stem cell population in culture condition and wide use to various types of stem cells. Human mesenchymal stem cell, adipose derived stem cell, tonsil derived stem cell and osteogenic differentiated cells migrated toward anode but the migration speed of differentiated cells was significantly decreased versus that of stem cells. In mixture of stem cells and differentiated cells condition, we identified that the ratio of stem cell versus differentiated cell was matched with the homogeneity evaluation data of stem cells based on electrotaxis analysis. As a result, our evaluation tool has the possibility of the wide use to stem cell homogeneity evaluation and might be used as the stem cell quality control during stem cell culture without any additional antibodies.
Methylseleninic Acid Provided at Nutritional Selenium Levels Inhibits Angiogenesis by Down-regulating Integrin β3 Signaling
Authors: Cai Zhihui , Dong Liangbo , Song Chengwei , Zhang Yanqing , Zhu Chenghui , Zhang Yibo , Ling Qinjie , Hoffmann Peter R. , Li Jun , Huang Zhi , Li Wei
Published: August 2017

Targeting angiogenesis has emerged as a promising strategy for cancer treatment. Methylseleninic acid (MSA) is a metabolite of selenium (Se) in animal cells that exhibits anti-oxidative and anti-cancer activities at levels exceeding Se nutritional requirements. However, it remains unclear whether MSA exerts its effects on cancer prevention by influencing angiogenesis within Se nutritional levels. Herein, we demonstrate that MSA inhibited angiogenesis at 2 µM, which falls in the range of moderate Se nutritional status. We found that MSA treatments at 2 µM increased cell adherence, while inhibiting cell migration and tube formation of HUVECs in vitro. Moreover, MSA effectively inhibited the sprouts of mouse aortic rings and neoangiogenesis in chick embryo chorioallantoic membrane. We also found that MSA down-regulated integrin β3 at the levels of mRNA and protein, and disrupted clustering of integrin β3 on the cell surface. Additionally, results showed that MSA inhibited the phosphorylation of AKT, IκBα, and NFκB. Overall, our results suggest that exogenous MSA inhibited angiogenesis at nutritional Se levels not only by down-regulating the expression of integrin β3 but also by disorganizing the clustering of integrin β3, which further inhibited the phosphorylation involving AKT, IκBα, NFκB. These findings provide novel mechanistic insight into the function of MSA for regulating angiogenesis and suggest that MSA could be a potential candidate or adjuvant for anti-tumor therapy in clinical settings.
Regulation of HGF-induced hepatocyte proliferation by the small GTPase Arf6 through the PIP2-producing enzyme PIP5K1A
Authors: Tsai Meng-Tsz , Katagiri Naohiro , Ohbayashi Norihiko , Iwasaki Kenichi , Ohkohchi Nobuhiro , Ding Shih-Torng , Kanaho Yasunori , Funakoshi Yuji
Published: August 2017

HGF and its receptor c-Met are critical molecules in various biological processes. Others and we have previously shown that the small GTPase Arf6 plays a pivotal role in HGF signaling in hepatocytes. However, the molecular mechanism of how Arf6 regulates HGF signaling is unclear. Here, we show that Arf6 plays an important role in HGF-stimulated hepatocyte proliferation and liver regeneration through the phosphatidylinositol 4,5-bisphosphate (PIP2)-producing enzyme PIP5K1A. We find that knockdown of Arf6 and PIP5K1A in HepG2 cells inhibits HGF-stimulated proliferation, Akt activation, and generation of phosphatidylinositol 3,4,5-trisphosphate (PIP3) and its precursor PIP2. Interestingly, PIP5K1A is recruited to c-Met upon HGF stimulation in an Arf6 activity-dependent manner. Finally, we show that hepatocyte proliferation and liver regeneration after partial hepatectomy are suppressed in Pip5k1a knockout mice. These results provide insight into the molecular mechanism for HGF-stimulated hepatocyte proliferation and liver regeneration: Arf6 recruits PIP5K1A to c-Met and activates it upon HGF stimulation to produce PIP2 and subsequently PIP3, which in turn activates Akt to promote hepatocyte proliferation, thereby accelerating liver regeneration after liver injury.
Inhibiting mycobacterial tryptophan synthase by targeting the inter-subunit interface
Authors: Abrahams Katherine A. , Cox Jonathan A. G. , Fütterer Klaus , Rullas Joaquín , Ortega-Muro Fátima , Loman Nicholas J. , Moynihan Patrick J. , Pérez-Herrán Esther , Jiménez Elena , Esquivias Jorge , Barros David , Ballell Lluís , Alemparte Carlos , Besra Gurdyal S.
Published: August 2017

Drug discovery efforts against the pathogen Mycobacterium tuberculosis (Mtb) have been advanced through phenotypic screens of extensive compound libraries. Such a screen revealed sulfolane 1 and indoline-5-sulfonamides 2 and 3 as potent inhibitors of mycobacterial growth. Optimization in the sulfolane series led to compound 4, which has proven activity in an in vivo murine model of Mtb infection. Here we identify the target and mode of inhibition of these compounds based on whole genome sequencing of spontaneous resistant mutants, which identified mutations locating to the essential α- and β-subunits of tryptophan synthase. Over-expression studies confirmed tryptophan synthase as the biological target. Biochemical techniques probed the mechanism of inhibition, revealing the mutant enzyme complex incurs a fitness cost but does not prevent inhibitor binding. Mapping of the resistance conferring mutations onto a low-resolution crystal structure of Mtb tryptophan synthase showed they locate to the interface between the α- and β-subunits. The discovery of anti-tubercular agents inhibiting tryptophan synthase highlights the therapeutic potential of this enzyme and draws attention to the prospect of other amino acid biosynthetic pathways as future Mtb drug targets.
Sost deficiency led to a greater cortical bone formation response to mechanical loading and altered gene expression
Authors: Pflanz David , Birkhold Annette I. , Albiol Laia , Thiele Tobias , Julien Catherine , Seliger Anne , Thomson Erin , Kramer Ina , Kneissel Michaela , Duda Georg N. , Kornak Uwe , Checa Sara , Willie Bettina M.
Published: August 2017

Bone adaptation optimizes mass and structure, but the mechano-response is already reduced at maturation. Downregulation of sclerostin was believed to be a mandatory step in mechano-adaptation, but in young mice it was shown that load-induced formation can occur independent of sclerostin, a product of the Sost gene. We hypothesized that the bone formation and resorption response to loading is not affected by Sost deficiency, but is age-specific. Our findings indicate that the anabolic response to in vivo tibial loading was reduced at maturation in Sost Knockout (KO) and littermate control (LC) mice. Age affected all anabolic and catabolic parameters and altered Sost and Wnt target gene expression. While load-induced cortical resorption was similar between genotypes, loading-induced gains in mineralizing surface was enhanced in Sost KO compared to LC mice. Loading led to a downregulation in expression of the Wnt inhibitor Dkk1. Expression of Dkk1 was greater in both control and loaded limbs of Sost KO compared to LC mice suggesting a compensatory role in the absence of Sost. These data suggest physical activity could enhance bone mass concurrently with sclerostin-neutralizing antibodies, but treatment strategies should consider the influence of age on ultimate load-induced bone mass gains.
Small RNA profiles from Panax notoginseng roots differing in sizes reveal correlation between miR156 abundances and root biomass levels
Authors: Zheng Yun , Chen Kun , Xu Zhenning , Liao Peiran , Zhang Xiaotuo , Liu Li , Wei Kangning , Liu Diqiu , Li Yong-Fang , Sunkar Ramanjulu , Cui Xiuming
Published: August 2017

Plant genomes encode several classes of small regulatory RNAs (sRNAs) that play critical roles in both development and stress responses. Panax notoginseng (Burk.) F.H. Chen (P. notoginseng) is an important traditional Chinese herbal medicinal plant species for its haemostatic effects. Therefore, the root yield of P. notoginseng is a major economically important trait since the roots of P. notoginseng are the parts used to produce medicine. To identify sRNAs that are critical for the root biomass of P. notoginseng, we performed a comprehensive study of miRNA transcriptomes from P. notoginseng roots of different biomasses. We identified 675 conserved miRNAs, of which 180 pre-miRNAs are also identified, and three TAS3 loci in P. notoginseng. By using degradome sequencing, we identified 79 conserved miRNA:target or tasiRNA:target interactions, of which eight were further confirmed with the RLM 5′-RACE experiments. More importantly, our results revealed that a member of miR156 family and one of its SPL target genes have inverse expression levels, which is tightly correlated with greater root biomass contents. These results not only contributes to overall understanding of post-transcriptional gene regulation in roots of P. notoginseng but also could serve as markers for breeding P. notoginseng with greater root yield.
Perfused Three-dimensional Organotypic Culture of Human Cancer Cells for Therapeutic Evaluation
Authors: Wan Xiao , Ball Steven , Willenbrock Frances , Yeh Shaoyang , Vlahov Nikola , Koennig Delia , Green Marcus , Brown Graham , Jeyaretna Sanjeeva , Li Zhaohui , Cui Zhanfeng , Ye Hua , O’Neill Eric
Published: August 2017

Pharmaceutical research requires pre-clinical testing of new therapeutics using both in-vitro and in-vivo models. However, the species specificity of non-human in-vivo models and the inadequate recapitulation of physiological conditions in-vitro are intrinsic weaknesses. Here we show that perfusion is a vital factor for engineered human tissues to recapitulate key aspects of the tumour microenvironment. Organotypic culture and human tumour explants were allowed to grow long-term (14–35 days) and phenotypic features of perfused microtumours compared with those in the static culture. Differentiation status and therapeutic responses were significantly different under perfusion, indicating a distinct biological response of cultures grown under static conditions. Furthermore, heterogeneous co-culture of tumour and endothelial cells demonstrated selective cell-killing under therapeutic perfusion versus episodic delivery. We present a perfused 3D microtumour culture platform that sustains a more physiological tissue state and increased viability for long-term analyses. This system has the potential to tackle the disadvantages inherit of conventional pharmaceutical models and is suitable for precision medicine screening of tumour explants, particularly in hard-to-treat cancer types such as brain cancer which suffer from a lack of clinical samples.
Serum fibronectin distinguishes the early stages of hepatocellular carcinoma
Authors: Kim Hyunsoo , Park JiYoung , Kim Yongkang , Sohn Areum , Yeo Injun , Jong Yu Su , Yoon Jung-Hwan , Park Taesung , Kim Youngsoo
Published: August 2017

Hepatocellular carcinoma (HCC) is the third leading cause of cancer-related death, necessitating the discovery of serum markers for its early detection. In this study, a total of 180 serum samples from liver cirrhosis (LC), hepatocellular carcinoma (HCC) patients and paired samples of HCC patients who recovered (Recovery) were analyzed by multiple reaction monitoring-mass spectrometry (MRM-MS) to verify biomarkers. The three-fold crossvalidation was repeated 100 times in the training and test sets to evaluate statistical significance of 124 candidate proteins. This step resulted in 2 proteins that had an area under the receiver operating curve (AUROC) values ≥0.800 in the training (n = 90) and test sets (n = 90). Specifically, fibronectin (FN1, WCGTTQNYDADQK), distinguished HCC from LC patients, with an AUROC value of 0.926 by logistic regression. A FN1 protein was selected for validation in an independent sample (n = 60) using enzyme-linked immunosorbent assay (ELISA). The combination of alpha-fetoprotein (AFP) and FN1 improved the diagnostic performance and differentiated HCC patients with normal AFP levels. Our study has examined candidate markers for the benign disease state and malignancy and has followed up on the consequent recovery. Thus, improvement in the early detection of HCC by a 2-marker panel (AFP + FN1) might benefit HCC patients.
A novel ruthenium (II)-derived organometallic compound, TQ-6, potently inhibits platelet aggregation: Ex vivo and in vivo studies
Authors: Hsia Chih-Hsuan , Velusamy Marappan , Sheu Joen-Rong , Khamrang Themmila , Jayakumar Thanasekaran , Lu Wan-Jung , Lin Kuan-Hung , Chang Chao-Chien
Published: August 2017

Arterial thrombosis plays a key role in cardiovascular diseases. Hence, developing more effective antithrombotic agents is necessary. We designed a ruthenium (II)-derived complex, [Ru(η6-cymene)2-(1H-benzoimidazol-2-yl)-quinoline Cl]BF4 (TQ-6), as a new antiplatelet drug. TQ-6 (0.3 µM) exhibited extremely strong inhibitory activity against platelet aggregation, Src, and Syk phosphorylation stimulated by agonists in human platelets. In collagen-activated platelets, TQ-6 also inhibited ATP-release, [Ca+2]i, P-selectin expression, FITC-PAC-1 binding, and hydroxyl radical formation, as well as the phosphorylation of phospholipase Cγ2, protein kinase C, mitogen-activated protein kinases, and Akt. Neither FITC-JAQ1 nor FITC-triflavin binding or integrin β3 phosphorylation stimulated by immobilized fibrinogen were diminished by TQ-6. Furthermore, TQ-6 had no effects in cyclic nucleotide formation. Moreover, TQ-6 substantially prolonged the closure time in whole blood, increased the occlusion time of thrombotic platelet plug formation and bleeding time in mice. In conclusion, TQ-6 has a novel role in inhibiting platelet activation through the inhibition of the agonist receptors-mediated inside-out signaling such as Src-Syk-PLCγ2 cascade and subsequent suppression of granule secretion, leading to disturb integrin αIIbβ3-mediated outside-in signaling, and ultimately inhibiting platelet aggregation. Therefore, TQ-6 has potential to develop as a therapeutic agent for preventing or treating thromboembolic disorders.
Circulating endothelial cell-derived extracellular vesicles mediate the acute phase response and sickness behaviour associated with CNS inflammation
Authors: Couch Yvonne , Akbar Naveed , Roodselaar Jay , Evans Matthew C. , Gardiner Chris , Sargent Ian , Romero Ignacio A. , Bristow Adrian , Buchan Alastair M. , Haughey Norman , Anthony Daniel C.
Published: August 2017

Brain injury elicits a systemic acute-phase response (APR), which is responsible for co-ordinating the peripheral immunological response to injury. To date, the mechanisms responsible for signalling the presence of injury or disease to selectively activate responses in distant organs were unclear. Circulating endogenous extracellular vesicles (EVs) are increased after brain injury and have the potential to carry targeted injury signals around the body. Here, we examined the potential of EVs, isolated from rats after focal inflammatory brain lesions using IL-1β, to activate a systemic APR in recipient naïve rats, as well as the behavioural consequences of EV transfer. Focal brain lesions increased EV release, and, following isolation and transfer, the EVs were sequestered by the liver where they initiated an APR. Transfer of blood-borne EVs from brain-injured animals was also enough to suppress exploratory behaviours in recipient naïve animals. EVs derived from brain endothelial cell cultures treated with IL-1β also activated an APR and altered behaviour in recipient animals. These experiments reveal that inflammation-induced circulating EVs derived from endothelial cells are able to initiate the APR to brain injury and are sufficient to generate the associated sickness behaviours, and are the first demonstration that EVs are capable of modifying behavioural responses.
Neuroprotective effect of kinin B1 receptor activation in acute cerebral ischemia in diabetic mice
Authors: Desposito Dorinne , Zadigue Georges , Taveau Christopher , Adam Clovis , Alhenc-Gelas François , Bouby Nadine , Roussel Ronan
Published: August 2017

Activation of the kallikrein-kinin system enhances cardiac and renal tolerance to ischemia. Here we investigated the effects of selective agonists of kinin B1 or B2 receptor (R) in brain ischemia-reperfusion in diabetic and non-diabetic mice. The role of endogenous kinins was assessed in tissue kallikrein deficient mice (TK−/−). Mice underwent 60min-middle cerebral artery occlusion (MCAO), eight weeks after type 1-diabetes induction. Treatment with B1R-, B2R-agonist or saline was started at reperfusion. Neurological deficit (ND), infarct size (IS), brain water content (BWC) were measured at day 0, 1 and 2 after injury. MCAO induced exaggerated ND, mortality and IS in diabetic mice. B2R-agonist increased ND and mortality to 60% and 80% in non-diabetic and diabetic mice respectively, by mechanisms involving hemodynamic failure and renal insufficiency. TK−/− mice displayed reduced ND and IS compared to wild-type littermate, consistent with suppression of B2R activity. B1R mRNA level increased in ischemic brain but B1R-agonist had no effect on ND, mortality or IS in non-diabetic mice. In contrast, in diabetic mice, B1R-agonist tested at two doses significantly reduced ND by 42–52% and IS by 66–71%, without effect on BWC or renal function. This suggests potential therapeutic interest of B1R agonism for cerebral protection in diabetes.
Hyperspectral data processing improves PpIX contrast during fluorescence guided surgery of human brain tumors
Authors: Bravo J. J. , Olson J. D. , Davis S. C. , Roberts D. W. , Paulsen K. D. , Kanick S. C.
Published: August 2017

Fluorescence guided surgery (FGS) using aminolevulinic-acid (ALA) induced protoporphyrin IX (PpIX) provides intraoperative visual contrast between normal and malignant tissue during resection of high grade gliomas. However, maps of the PpIX biodistribution within the surgical field based on either visual perception or the raw fluorescence emissions can be masked by background signals or distorted by variations in tissue optical properties. This study evaluates the impact of algorithmic processing of hyperspectral imaging acquisitions on the sensitivity and contrast of PpIX maps. Measurements in tissue-simulating phantoms showed that (I) spectral fitting enhanced PpIX sensitivity compared with visible or integrated fluorescence, (II) confidence-filtering automatically determined the lower limit of detection based on the strength of the PpIX spectral signature in the collected emission spectrum (0.014–0.041 μg/ml in phantoms), and (III) optical-property corrected PpIX estimates were more highly correlated with independent probe measurements (r = 0.98) than with spectral fitting alone (r = 0.91) or integrated fluorescence (r = 0.82). Application to in vivo case examples from clinical neurosurgeries revealed changes to the localization and contrast of PpIX maps, making concentrations accessible that were not visually apparent. Adoption of these methods has the potential to maintain sensitive and accurate visualization of PpIX contrast over the course of surgery.