bluuebunny/crossref_metadata_2025 huggingface dataset:
Increased clusterin levels after myocardial infarction is due to a defect in protein degradation systems activity
Authors: Turkieh Annie , Porouchani Sina , Beseme Olivia , Chwastyniak Maggy , Amouyel Philippe , Lamblin Nicolas , Balligand Jean-Luc , Bauters Christophe , Pinet Florence
Published: August 2019

Clusterin (CLU) is induced in many organs after tissue injury or remodeling. Recently, we show that CLU levels are increased in plasma and left ventricle (LV) after MI, however, the mechanisms involved are not yet elucidated. On the other hand, it has been shown that the activity of the protein degradation systems (PDS) is affected after MI with a decrease in ubiquitin proteasome system (UPS) and an increase in macroautophagy. The aim of this study was to decipher if the increased CLU levels after MI are in part due to the alteration of PDS activity. Rat neonate cardiomyocytes (NCM) were treated with different modulators of UPS and macroautophagy in order to decipher their role in CLU expression, secretion, and degradation. We observed that inhibition of UPS activity in NCM increased CLU mRNA levels, its intracellular protein levels (p-CLU and m-CLU) and its secreted form (s-CLU). Macroautophagy was also induced after MG132 treatment but is not active. The inhibition of macroautophagy induction in MG132-treated NCM increased CLU mRNA and m-CLU levels, but not s-CLU compared to NCM only treated by MG132. We also demonstrate that CLU can be degraded in NCM through proteasome and lysosome by a macroautophagy independent pathway. In another hand, CLU silencing in NCM has no effect either on macroautophagy or apoptosis induced by MG132. However, the overexpression of CLU secreted isoform in H9c2 cells, but not in NCM decreased apoptosis after MG132 treatment. Finally, we observed that increased CLU levels in hypertrophied NCM and in failing human hearts are associated with proteasome inhibition and macroautophagy alteration. All these data suggest that increased CLU expression and secretion after MI is, in part, due to a defect of UPS and macroautophagy activities in the heart and may have a protective effect by decreasing apoptosis induced by proteasome inhibition.
Mild hypothermia improves neurological outcome in mice after cardiopulmonary resuscitation through Silent Information Regulator 1-actviated autophagy
Authors: Wei Hongyan , Yin Meixian , Lu Yuanzheng , Yang Yan , Li Bo , Liao Xiao-Xing , Dai Gang , Jing Xiaoli , Xiong Yan , Hu Chunlin
Published: August 2019

Mild hypothermia treatment (MHT) improves the neurological function of cardiac arrest (CA) patients, but the exact mechanisms of recovery remain unclear. Herein, we generated a CA and cardiopulmonary resuscitation (CPR) mouse model to elucidate such function. Naïve mice were randomly divided into two groups, a normothemia (NT) group, in which animals had normal body temperature, and a MHT group, in which animals had a body temperature of 33 °C (range: 32–34 °C), after the return of spontaneous circulation (ROSC), followed by CA/CPR. MHT significantly improved the survival rate of CA/CPR mice compared with NT. Mechanistically, MHT increased the expression of Silent Information Regulator 1 (Sirt1) and decreased P53 phosphorylation (p-P53) in the cortex of CA/CPR mice, which coincided with the elevated autophagic flux. However, Sirt1 deletion compromised the neuroprotection offered by MHT, indicating that Sirt1 plays an important role. Consistent with the observations obtained from in vivo work, our in vitro study utilizing cultured neurons subjected to oxygen/glucose deprivation and reperfusion (OGD/R) also indicated that Sirt1 knockdown increased OGD/R-induced neuron necrosis and apoptosis, which was accompanied by decreased autophagic flux and increased p-P53. However, the depletion of P53 did not suppress neuron death, suggesting that P53 was not critically involved in MHT-induced neuroprotection. In contrast, the application of autophagic inhibitor 3-methyladenine attenuated MHT-improved neuron survival after OGD/R, further demonstrating that increased autophagic flux significantly contributes to MHT-linked neuroprotection of CA/CRP mice. Our findings indicate that MHT improves neurological outcome of mice after CA/CPR through Sirt1-mediated activation of autophagic flux.
Band-collision gel electrophoresis
Authors: Bikos Dimitri A. , Mason Thomas G.
Published: August 2019

Electrophoretic mobility shift assays are widely used in gel electrophoresis to study binding interactions between different molecular species loaded into the same well. However, shift assays can access only a subset of reaction possibilities that could be otherwise seen if separate bands of reagent species might instead be collisionally reacted. Here, we adapt gel electrophoresis by fabricating two or more wells in the same lane, loading these wells with different reagent species, and applying an electric field, thereby producing collisional reactions between propagating pulse-like bands of these species, which we image optically. For certain pairs of anionic and cationic dyes, propagating bands pass through each other unperturbed; yet, for other pairs, we observe complexing and precipitation reactions, indicating strong attractive interactions. We generalize this band-collision gel electrophoresis (BCGE) approach to other reaction types, including acid-base, ligand exchange, and redox, as well as to colloidal species in passivated large-pore gels.
Towards general network architecture design criteria for negative gas adsorption transitions in ultraporous frameworks
Authors: Krause Simon , Evans Jack D. , Bon Volodymyr , Senkovska Irena , Iacomi Paul , Kolbe Felicitas , Ehrling Sebastian , Troschke Erik , Getzschmann Jürgen , Többens Daniel M. , Franz Alexandra , Wallacher Dirk , Yot Pascal G. , Maurin Guillaume , Brunner Eike , Llewellyn Philip L. , Coudert François-Xavier , Kaskel Stefan
Published: August 2019

Switchable metal-organic frameworks (MOFs) have been proposed for various energy-related storage and separation applications, but the mechanistic understanding of adsorption-induced switching transitions is still at an early stage. Here we report critical design criteria for negative gas adsorption (NGA), a counterintuitive feature of pressure amplifying materials, hitherto uniquely observed in a highly porous framework compound (DUT-49). These criteria are derived by analysing the physical effects of micromechanics, pore size, interpenetration, adsorption enthalpies, and the pore filling mechanism using advanced in situ X-ray and neutron diffraction, NMR spectroscopy, and calorimetric techniques parallelised to adsorption for a series of six isoreticular networks. Aided by computational modelling, we identify DUT-50 as a new pressure amplifying material featuring distinct NGA transitions upon methane and argon adsorption. In situ neutron diffraction analysis of the methane (CD4) adsorption sites at 111 K supported by grand canonical Monte Carlo simulations reveals a sudden population of the largest mesopore to be the critical filling step initiating structural contraction and NGA. In contrast, interpenetration leads to framework stiffening and specific pore volume reduction, both factors effectively suppressing NGA transitions.
Engineering a microbial biosynthesis platform for de novo production of tropane alkaloids
Authors: Srinivasan Prashanth , Smolke Christina D.
Published: August 2019

Tropane alkaloids (TAs) are a class of phytochemicals produced by plants of the nightshade family used for treating diverse neurological disorders. Here, we demonstrate de novo production of tropine, a key intermediate in the biosynthetic pathway of medicinal TAs such as scopolamine, from simple carbon and nitrogen sources in yeast (Saccharomyces cerevisiae). Our engineered strain incorporates 15 additional genes, including 11 derived from diverse plants and bacteria, and 7 disruptions to yeast regulatory or biosynthetic proteins to produce tropine at titers of 6 mg/L. We also demonstrate the utility of our engineered yeast platform for the discovery of TA derivatives by combining biosynthetic modules from distant plant lineages to achieve de novo production of cinnamoyltropine, a non-canonical TA. Our engineered strain constitutes a starting point for future optimization efforts towards realizing industrial fermentation of medicinal TAs and a platform for the synthesis of TA derivatives with enhanced bioactivities.
Parallels in the sequential organization of birdsong and human speech
Authors: Sainburg Tim , Theilman Brad , Thielk Marvin , Gentner Timothy Q.
Published: August 2019

Human speech possesses a rich hierarchical structure that allows for meaning to be altered by words spaced far apart in time. Conversely, the sequential structure of nonhuman communication is thought to follow non-hierarchical Markovian dynamics operating over only short distances. Here, we show that human speech and birdsong share a similar sequential structure indicative of both hierarchical and Markovian organization. We analyze the sequential dynamics of song from multiple songbird species and speech from multiple languages by modeling the information content of signals as a function of the sequential distance between vocal elements. Across short sequence-distances, an exponential decay dominates the information in speech and birdsong, consistent with underlying Markovian processes. At longer sequence-distances, the decay in information follows a power law, consistent with underlying hierarchical processes. Thus, the sequential organization of acoustic elements in two learned vocal communication signals (speech and birdsong) shows functionally equivalent dynamics, governed by similar processes.
(Pro)renin receptor contributes to renal mitochondria dysfunction, apoptosis and fibrosis in diabetic mice
Authors: Li Caixia , Matavelli Luis C. , Akhtar Safia , Siragy Helmy M.
Published: August 2019

Recently we demonstrated that increased renal (Pro)renin receptor (PRR) expression in diabetes contributes to development of diabetic kidney disease. However, the exact mechanisms involving PRR activity and diabetic kidney dysfunction are unknown. We hypothesized that PRR is localized in renal mitochondria and contributes to renal fibrosis and apoptosis through oxidative stress-induced mitochondria dysfunction. Controls and streptozotocin-induced diabetic C57BL/6 mice were injected with scramble shRNA and PRR shRNA and followed for a period of eight weeks. At the end of study, diabetic mice showed increased expressions of PRR and NOX4 in both total kidney tissue and renal mitochondria fraction. In addition, renal mitochondria of diabetic mice showed reduced protein expression and activity of SOD2 and ATP production and increased UCP2 expression. In diabetic kidney, there was upregulation in the expressions of caspase3, phos-Foxo3a, phos-NF-κB, fibronectin, and collagen IV and reduced expressions of Sirt1 and total-FOXO3a. Renal immunostaining revealed increased deposition of PRR, collagen and fibronectin in diabetic kidney. In diabetic mice, PRR knockdown decreased urine albumin to creatinine ratio and the renal expressions of PRR, NOX4, UCP2, caspase3, phos-FOXO3a, phos-NF-κB, collagen, and fibronectin, while increased the renal mitochondria expression and activity of SOD2, ATP production, and the renal expressions of Sirt1 and total-FOXO3a. In conclusion, increased expression of PRR localized in renal mitochondria and diabetic kidney induced mitochondria dysfunction, and enhanced renal apoptosis and fibrosis in diabetes by upregulation of mitochondria NOX4/SOD2/UCP2 signaling pathway.
Assessment of intracardiac and extracardiac anomalies associated with coarctation of aorta and interrupted aortic arch using dual-source computed tomography
Authors: Zhao Qin , Wang Jin , Yang Zhi-gang , Shi Ke , Diao Kai-yue , Huang Shan , Shen Meng-ting , Guo Ying-kun
Published: August 2019

To evaluate the value of dual-source computed tomography (DSCT) compared with transthoracic echocardiography (TTE) in assessing intracardiac and extracardiac anomalies in patients with coarctation of aorta (CoA) and interrupted aortic arch (IAA). Seventy-five patients (63 with CoA and 12 with IAA) who received preoperative DSCT and TTE were retrospectively studied. Intracardiac and extracardiac anomalies were recorded and compared by DSCT and TTE, in reference to surgical or cardiac catheterization findings. A total of 155 associated anomalies were finally found. Collateral circulation (56, 74.70%), patent ductus arteriosus (PDA; 41, 54.67%) were the most common anomalies. PDA, aortopulmonary window, and collateral circulation were more frequently present in patients with IAA than those with CoA (100% vs. 46.03%, 16.67% vs. 0%, and 100% vs. 69.84%, respectively, all p < 0.05). DSCT was superior to TTE in assessing associated extracardiac-vascular anomalies (sensitivity: 100% vs. 39.81%; specificity: 100% vs. 100%; positive predictive value: 100% vs. 100%; negative predictive value: 100% vs. 76.06%). Extracardiac-vascular anomalies, including collateral circulation and PDA, were the most common anomalies in patients with IAA and CoA. Compared with TTE, DSCT is more reliable in providing an overall preoperative evaluation of morphological features and extracardiac anomalies for surgical planning.
VEGF mediates fat embolism-induced acute lung injury via VEGF receptor 2 and the MAPK cascade
Authors: Lin Chin-Kuo , Lin Yu-Hao , Huang Tai-Chun , Shi Chung-Sheng , Yang Cheng-Ta , Yang Yi-Ling
Published: August 2019

Fat embolism (FE) is a lethal medical emergency often caused by fracture of long bones and amputation of limbs. Vascular endothelial growth factor (VEGF) promotes angiogenesis and increases vascular permeability. We tested the hypothesis that VEGF plays a critical role in FE-induced acute respiratory distress syndrome (ARDS) and acute lung injury (ALI). Fat tissues were collected from male Sprague-Dawley rats, and animal oil was extracted and mixed with water to form fatty micelles. The micelles were then injected into the tail vein to produce FE and ALI in rats. Lung weight gain was measured as the index of pulmonary edema. The expression of pulmonary VEGF was evaluated by real-time PCR and western blot analysis. Inducible nitric oxide synthase (iNOS) and phosphorylation of mitogen-activated protein kinase (MAPK) were determined by western blot analyses. Interleukin-1β (IL-1β) was quantified by ELISAs. Hematoxylin and eosin staining was used to evaluate the pathological damage of ALI. In this study, we found that animal oil-induced FE significantly increased pulmonary VEGF expression and MAPK phosphorylation. We also evaluated the inflammatory response after FE and found that iNOS and IL-1β significantly increased after FE. Systemic administration of SU-1498, an antagonist of VEGF receptor 2 (VEGFR-2), significantly attenuated the FE-induced inflammatory response and histological damage. This study suggested that VEGF is involved in FE-induced ARDS via the VEGFR-2 and MAPK cascades, which induce IL-1β release and iNOS upregulation. Blockade of could be used to treat FE-induced pulmonary damage.
Involvement of the P2X7 receptor in the migration and metastasis of tamoxifen-resistant breast cancer: effects on small extracellular vesicles production
Authors: Park Miso , Kim Jieun , Phuong Nguyen T. T. , Park Jung Gyu , Park Jin-Hee , Kim Yong-Chul , Baek Moon Chang , Lim Sung Chul , Kang Keon Wook
Published: August 2019

Tamoxifen (TAM) is the standard anti-hormonal therapy for estrogen receptor-positive breast cancer. However, long-term TAM therapy can make acquisition of TAM resistance and there are still no solutions to treat TAM-resistant breast cancer. In this study, we found that protein and mRNA expression of the P2X purinoreceptor 7 (P2X7) was higher in tamoxifen resistant MCF-7 (TAMR-MCF-7) cells than in control MCF-7 cells. P2X7 inhibition potently inhibited the migration of TAMR-MCF-7 cells and the liver metastasis burden of TAMR-MCF-7 cells in the spleen-liver metastasis experiment. However, the P2X7 antagonist did not affect protein expression of matrix metalloproteinase (MMP)-2, MMP-9, and epithelial-mesenchymal transition markers. Here our data indicate a link between small extracellular vesicles (sEV) and P2X7, and suggest a new mechanism of metastasis in TAM-resistant breast cancer cells through P2X7 receptors. The migration of TAMR-MCF-7 cells was increased in a concentration-dependent manner by purified sEV treatment. The number of secreted sEVs and the protein levels of CD63 in TAMR-MCF-7 cells were decreased by the P2X7 antagonist, showing that P2X7 influences the production of sEV. Our results suggest that inhibiting the P2X7 could be considered for metastasis prevention in TAM-resistant cancer patients.
Exploring the initiation of fiber fuse
Authors: Xiao Qirong , Tian Jiading , Yan Ping , Li Dan , Gong Mali
Published: August 2019

We report an investigation of conditions for the initiation of fiber fuse (IFF), a kind of catastrophic damage that troubles all kinds of optical fibers, in silica-based optical fibers. The fibers of different chemical compositions were processed and tested in controlled conditions without mechanical damages before the IFF. For all the fibers of IFF, the same correlation between the critical temperatures and the optical powers transmitted therein was revealed for the first time. The fibers of different chemical compositions exhibited different resistances to the IFF under the threshold powers for propagation of fiber fuses. The results offered promise for predicting fiber fuses in optical fiber systems, which could facilitate avoiding catastrophic losses. They could direct the optimization of fiber production technologies for suppressing the damages, as well as open a new path towards controlled utilization of fiber fuse in in-fiber microstructure fabrication.
OsCAldOMT1 is a bifunctional O-methyltransferase involved in the biosynthesis of tricin-lignins in rice cell walls
Authors: Lam Pui Ying , Tobimatsu Yuki , Matsumoto Naoyuki , Suzuki Shiro , Lan Wu , Takeda Yuri , Yamamura Masaomi , Sakamoto Masahiro , Ralph John , Lo Clive , Umezawa Toshiaki
Published: August 2019

Lignin is a phenylpropanoid polymer produced in the secondary cell walls of vascular plants. Although most eudicot and gymnosperm species generate lignins solely via polymerization of p-hydroxycinnamyl alcohols (monolignols), grasses additionally use a flavone, tricin, as a natural lignin monomer to generate tricin-incorporated lignin polymers in cell walls. We previously found that disruption of a rice 5-HYDROXYCONIFERALDEHYDE O-METHYLTRANSFERASE (OsCAldOMT1) reduced extractable tricin-type metabolites in rice vegetative tissues. This same enzyme has also been implicated in the biosynthesis of sinapyl alcohol, a monolignol that constitutes syringyl lignin polymer units. Here, we further demonstrate through in-depth cell wall structural analyses that OsCAldOMT1-deficient rice plants produce altered lignins largely depleted in both syringyl and tricin units. We also show that recombinant OsCAldOMT1 displayed comparable substrate specificities towards both 5-hydroxyconiferaldehyde and selgin intermediates in the monolignol and tricin biosynthetic pathways, respectively. These data establish OsCAldOMT1 as a bifunctional O-methyltransferase predominantly involved in the two parallel metabolic pathways both dedicated to the biosynthesis of tricin-lignins in rice cell walls. Given that cell wall digestibility was greatly enhanced in the OsCAldOMT1-deficient rice plants, genetic manipulation of CAldOMTs conserved in grasses may serve as a potent strategy to improve biorefinery applications of grass biomass.
TGR(mREN2)27 rats develop non-alcoholic fatty liver disease-associated portal hypertension responsive to modulations of Janus-kinase 2 and Mas receptor
Authors: Klein Sabine , Kleine Carola-Ellen , Pieper Andrea , Granzow Michaela , Gautsch Sebastian , Himmit Mimoun , Kahrmann Katharina , Schierwagen Robert , Uschner Frank Erhard , Magdaleno Fernando , Naoum Maria Eleni , Kristiansen Glen , Walther Thomas , Bader Michael , Sauerbruch Tilman , Trebicka Jonel
Published: August 2019

Prevalence of non-alcoholic fatty liver disease (NAFLD) is increasing. Resulting fibrosis and portal hypertension, as a possible secondary event, may necessitate treatment. Overexpression of mouse renin in the transgenic rat model, TGR(mREN2)27, leads to spontaneous development of NAFLD. Therefore, we used TGR(mREN2)27 rats as a model of NAFLD where we hypothesized increased susceptibility and investigated fibrosis and portal hypertension and associated pathways. 12-week old TGR(mREN2)27 rats received either cholestatic (BDL) or toxic injury (CCl4 inhalation). Portal and systemic hemodynamic assessments were performed using microsphere technique with and without injection of the Janus-Kinase 2 (JAK2) inhibitor AG490 or the non-peptidic Ang(1-7) agonist, AVE0991. The extent of liver fibrosis was assessed in TGR(mREN2)27 and wild-type rats using standard techniques. Protein and mRNA levels of profibrotic, renin-angiotensin system components were assessed in liver and primary hepatic stellate cells (HSC) and hepatocytes. TGR(mREN2)27 rats developed spontaneous, but mild fibrosis and portal hypertension due to the activation of the JAK2/Arhgef1/ROCK pathway. AG490 decreased migration of HSC and portal pressure in isolated liver perfusions and in vivo. Fibrosis or portal hypertension after cholestatic (BDL) or toxic injury (CCl4) was not aggravated in TGR(mREN2)27 rats, probably due to decreased mouse renin expression in hepatocytes. Interestingly, portal hypertension was even blunted in TGR(mREN2)27 rats (with or without additional injury) by AVE0991. TGR(mREN2)27 rats are a suitable model of spontaneous liver fibrosis and portal hypertension but not with increased susceptibility to liver damage. After additional injury, the animals can be used to evaluate novel therapeutic strategies targeting Mas.
Opposing roles of endothelial and leukocyte-expressed IL-7Rα in the regulation of psoriasis-like skin inflammation
Authors: Vranova Martina , Friess Mona C. , Haghayegh Jahromi Neda , Collado-Diaz Victor , Vallone Angela , Hagedorn Olivia , Jadhav Maria , Willrodt Ann-Helen , Polomska Anna , Leroux Jean-Christophe , Proulx Steven T. , Halin Cornelia
Published: August 2019

The interleukin 7 receptor alpha chain (IL-7Rα) is predominately expressed by lymphocytes, and activation by its ligand IL-7 supports the development and maintenance of T cells and boosts T-cell mediated immunity. We recently reported that lymphatic endothelial cells (LECs) in dermal lymphatics also express IL-7 and its receptor chains (IL-7Rα and CD132) and that IL-7 supports lymphatic drainage. This suggested that activation of IL-7Rα signaling in lymphatics could exert inflammation-resolving activity, by promoting the clearance of excess tissue fluid. Here we investigated how the potentially opposing effects of IL-7Rα signaling in immune cells and in the lymphatic vasculature would affect the development and progression of psoriasis-like skin inflammation. We found that during acute and chronic skin inflammation mice with an endothelial-specific deletion of IL-7Rα (IL-7RαΔEC mice) developed more edema compared to control mice, as a consequence of impaired lymphatic drainage. However, systemic treatment of wild-type mice with IL-7 exacerbated edema and immune cell infiltration in spite of increasing lymphatic drainage, whereas treatment with IL-7Rα blocking antibody ameliorated inflammatory symptoms. These data identify IL-7Rα signaling as a new pathway in psoriasis-like skin inflammation and show that its pro-inflammatory effects on the immune compartment override its anti-inflammatory, drainage-enhancing effects on the endothelium.
Crowdsourcing punishment: Individuals reference group preferences to inform their own punitive decisions
Authors: Son Jae-Young , Bhandari Apoorva , FeldmanHall Oriel
Published: August 2019

Justice systems delegate punishment decisions to groups in the belief that the aggregation of individuals’ preferences facilitates judiciousness. However, group dynamics may also lead individuals to relinquish moral responsibility by conforming to the majority’s preference for punishment. Across five experiments (N = 399), we find Victims and Jurors tasked with restoring justice become increasingly punitive (by as much as 40%) as groups express a desire to punish, with every additional punisher augmenting an individual’s punishment rates. This influence is so potent that knowing about a past group’s preference continues swaying decisions even when they cannot affect present outcomes. Using computational models of decision-making, we test long-standing theories of how groups influence choice. We find groups induce conformity by making individuals less cautious and more impulsive, and by amplifying the value of punishment. However, compared to Victims, Jurors are more sensitive to moral violation severity and less readily swayed by the group. Conformity to a group’s punitive preference also extends to weightier moral violations such as assault and theft. Our results demonstrate that groups can powerfully shift an individual’s punitive preference across a variety of contexts, while additionally revealing the cognitive mechanisms by which social influence alters moral values.
Temporal attention causes systematic biases in visual confidence
Authors: Recht Samuel , Mamassian Pascal , de Gardelle Vincent
Published: August 2019

Temporal attention enhances the perceptual representation of a stimulus at a particular point in time. The number of possible attentional episodes in a given period is limited, but whether observers’ confidence reflects such limitations is still unclear. To investigate this issue, we adapted an “Attentional Blink” paradigm, presenting observers with a rapid visual stream of letters containing two targets cued for subsequent perceptual reports and confidence judgments. We found three main results. First, when two targets fell within the same attentional episode, the second target underwent a strong under-confidence bias. In other words, confidence neglected that a single attentional episode can benefit to both targets. Second, despite this initial bias, confidence was strongly correlated with response probability. Third, as confidence was yoked to the evidence used in perceptual reports, it remains blind to delays in response selection for the second target. Notably, the second target was often mistaken with a later item associated with higher confidence. These results suggest that confidence does not perfectly evaluate the limits of temporal attention in challenging situations.
Cancer Testis Antigen Promotes Triple Negative Breast Cancer Metastasis and is Traceable in the Circulating Extracellular Vesicles
Authors: Kannan Anbarasu , Philley Julie V. , Hertweck Kate L. , Ndetan Harrison , Singh Karan P. , Sivakumar Subramaniam , Wells Robert B. , Vadlamudi Ratna K. , Dasgupta Santanu
Published: August 2019

Triple negative breast cancer (TNBC) has poor survival, exhibits rapid metastases, lacks targeted therapies and reliable prognostic markers. Here, we examined metastasis promoting role of cancer testis antigen SPANXB1 in TNBC and its utility as a therapeutic target and prognostic biomarker. Expression pattern of SPANXB1 was determined using matched primary cancer, lymph node metastatic tissues and circulating small extracellular vesicles (sEVs). cDNA microarray analysis of TNBC cells stably integrated with a metastasis suppressor SH3GL2 identified SPANXB1 as a potential target gene. TNBC cells overexpressing SH3GL2 exhibited decreased levels of both SPANXB1 mRNA and protein. Silencing of SPANXB1 reduced migration, invasion and reactive oxygen species production of TNBC cells. SPANXB1 depletion augmented SH3GL2 expression and decreased RAC-1, FAK, A-Actinin and Vinculin expression. Phenotypic and molecular changes were reversed upon SPANXB1 re-expression. SPANXB1 overexpressing breast cancer cells with an enhanced SPANXB1:SH3GL2 ratio achieved pulmonary metastasis within 5 weeks, whereas controls cells failed to do so. Altered expression of SPANXB1 was detected in the sEVs of SPANXB1 transduced cells. Exclusive expression of SPANXB1 was traceable in circulating sEVs, which was associated with TNBC progression. SPANXB1 represents a novel and ideal therapeutic target for blocking TNBC metastases due to its unique expression pattern and may function as an EV based prognostic marker to improve TNBC survival. Uniquely restricted expression of SPANXB1 in TNBCs, makes it an ideal candidate for targeted therapeutics and prognostication.
In Utero Gene Therapy (IUGT) Using GLOBE Lentiviral Vector Phenotypically Corrects the Heterozygous Humanised Mouse Model and Its Progress Can Be Monitored Using MRI Techniques
Authors: Shangaris Panicos , Loukogeorgakis Stavros P. , Subramaniam Sindhu , Flouri Christina , Jackson Laurence H. , Wang Wei , Blundell Michael P. , Liu Shanrun , Eaton Simon , Bakhamis Nahla , Ramachandra Durrgah Latchumi , Maghsoudlou Panayiotis , Urbani Luca , Waddington Simon N. , Eddaoudi Ayad , Archer Joy , Antoniou Michael N. , Stuckey Daniel J. , Schmidt Manfred , Thrasher Adrian J. , Ryan Thomas M. , De Coppi Paolo , David Anna L.
Published: August 2019

In utero gene therapy (IUGT) to the fetal hematopoietic compartment could be used to treat congenital blood disorders such as β-thalassemia. A humanised mouse model of β-thalassemia was used, in which heterozygous animals are anaemic with splenomegaly and extramedullary hematopoiesis. Intrahepatic in utero injections of a β globin-expressing lentiviral vector (GLOBE), were performed in fetuses at E13.5 of gestation. We analysed animals at 12 and 32 weeks of age, for vector copy number in bone marrow, peripheral blood liver and spleen and we performed integration site analysis. Compared to noninjected heterozygous animals IUGT normalised blood haemoglobin levels and spleen weight. Integration site analysis showed polyclonality. The left ventricular ejection fraction measured using magnetic resonance imaging (MRI) in treated heterozygous animals was similar to that of normal non-β-thalassemic mice but significantly higher than untreated heterozygous thalassemia mice suggesting that IUGT ameliorated poor cardiac function. GLOBE LV-mediated IUGT normalised the haematological and anatomical phenotype in a heterozygous humanised model of β-thalassemia.
Vitamin D reverts resistance to the mTOR inhibitor everolimus in hepatocellular carcinoma through the activation of a miR-375/oncogenes circuit
Authors: Provvisiero Donatella Paola , Negri Mariarosaria , de Angelis Cristina , Di Gennaro Gilda , Patalano Roberta , Simeoli Chiara , Papa Fortuna , Ferrigno Rosario , Auriemma Renata Simona , De Martino Maria Cristina , Colao Annamaria , Pivonello Rosario , Pivonello Claudia
Published: August 2019

Primary or acquired resistant mechanisms prevent the employment of individualized therapy with target drugs like the mTOR inhibitor everolimus (EVE) in hepatocellular carcinoma (HCC). The current study evaluated the effect of 1,25(OH)2Vitamin D (VitD) treatment on EVE sensitivity in established models of HCC cell lines resistant to everolimus (EveR). DNA content and colony formation assays, which measure the proliferative index, revealed that VitD pre-treatment re-sensitizes EveR cells to EVE treatment. The evaluation of epithelial and mesenchymal markers by western blot and immunofluorescence showed that VitD restored an epithelial phenotype in EveR cells, in which prolonged EVE treatment induced transition to mesenchymal phenotype. Moreover, VitD treatment prompted hepatic miRNAs regulation, evaluated by liver miRNA finder qPCR array. In particular, miR-375 expression was up-regulated by VitD in EveR cells, in which miR-375 was down-regulated compared to parental cells, with consequent inhibition of oncogenes involved in drug resistance and epithelial-mesenchymal transition (EMT) such as MTDH, YAP-1 and c-MYC. In conclusion, the results of the current study demonstrated that VitD can re-sensitize HCC cells resistant to EVE treatment triggering miR-375 up-regulation and consequently down-regulating several oncogenes responsible of EMT and drug resistance.
Changes in aniseikonia and influencing-factors following successful macula-off retinal detachment surgery
Authors: Murakami Tomoya , Okamoto Fumiki , Sugiura Yoshimi , Okamoto Yoshifumi , Hiraoka Takahiro , Oshika Tetsuro
Published: August 2019

This study investigated the changes in the severity of aniseikonia after surgery for macula-off retinal detachment (RD), and the relationship between aniseikonia and retinal microstructures. The study included 26 eyes of 26 patients undergoing RD surgery. Visual acuity was measured preoperatively, and at 3, 6, and 12 months postoperatively. Degree of aniseikonia and OCT images were obtained at 3, 6, and 12 months postoperatively. The aniseikonia values (mean ± standard deviation) at 3, 6, and 12 months postoperatively were −5.3 ± 4.2%, −4.4 ± 4.4%, and −3.1 ± 3.2%, respectively. Significant improvement was observed from 3 to 12 months postoperatively (P = 0.001). Twelve months postoperatively, 14 eyes had micropsia, 1 eye had macropsia, and 11 eyes were free of aniseikonia. Stepwise multiple regression analyses revealed that the severity of aniseikonia at 12 months postoperatively was significantly associated with postoperative development of cystoid macular edema (CME) and epiretinal membrane (ERM), as well as area of preoperative RD. In conclusion, although aniseikonia was gradually relieved after RD surgery during a 1-year follow-up period, approximately half of patients had aniseikonia and almost all of them had micropsia. Aniseikonia was associated with presence of postoperative CME, ERM, and area of preoperative RD.
A common phytoene synthase mutation underlies white petal varieties of the California poppy
Authors: Pollack Andrew J. , Gong Xue , Pollack Jonathan R.
Published: August 2019

The California poppy (Eschscholzia californica) is renowned for its brilliant golden-orange flowers, though white petal variants have been described. By whole-transcriptome sequencing, we have discovered in multiple white petal varieties a single deletion leading to altered splicing and C-terminal truncation of phytoene synthase (PSY), a key enzyme in carotenoid biosynthesis. Our findings underscore the diverse roles of phytoene synthase in shaping horticultural traits, and resolve a longstanding mystery of the regaled golden poppy.
Community phylogenetic structure reveals the imprint of dispersal-related dynamics and environmental filtering by nutrient availability in freshwater diatoms
Authors: Keck François , Kahlert Maria
Published: August 2019

Despite important progress, uncertainty persists regarding the ecological forces driving microbial community assembly. Here, we present the first study to use phylogenetic information to interpret the structure and diversity of diatom communities. We examined local phylogenetic divergence and beta- phylogenetic diversity in a large dataset of 595 freshwater benthic diatom communities and we investigated how this diversity is influenced by gradients in nutrients, pH, organic matter and catchment size. Overall, we found that diatom communities were phylogenetically clustered, i.e. species within communities were more closely related than expected by chance. Phylogenetic clustering was stronger in nutrient-poor environments and in sites with a small catchment area. The variation of the phylogenetic beta-diversity index was much better explained by space and environment than the variation of the taxonomic index was. Both approaches detected a significant effect of environment and space on diatom community turnover. Our results support the view that diatom communities are primarily shaped by environmental filtering, in particular by nutrient availability. Moreover, they highlight the importance of considering dispersal-related processes and the depth of phylogenetic signal in functional traits when interpreting patterns of diversity.
Three dimensional structures of putative, primitive proteins to investigate the origin of homochirality
Authors: Oda Akifumi , Nakayoshi Tomoki , Kato Koichi , Fukuyoshi Shuichi , Kurimoto Eiji
Published: August 2019

Primitive proteins are likely to have been constructed from non-enzymatically generated amino acids, due to the weak enzymatic activities of primitive biomolecules such as ribozymes. On the other hand, almost all present proteins are constructed only from l-amino acids. Therefore, there must have been a mechanism early in the origins of life that selected for one of the optical isomers of amino acids. In this study, we used molecular dynamics simulations to predict the three-dimensional structures of the putative primitive proteins constructed only from glycine, alanine, aspartic acid, and valine ([GADV]-peptides). The [GADV]-peptides were generated computationally at random from l-amino acids (l-[GADV]-peptides) and from both l- and d-amino acids (dl-[GADV]-peptides). The results indicate that the tendency of secondary structure formation for l-[GADV]-peptides was larger than that for dl-[GADV]-peptides, and l-[GADV]-peptides were more rigid than dl-[GADV]-peptides. These results suggest that the proteins with rigid structure motifs were more prone to have been generated in a primordial soup that included only l-amino acids than a the soup including racemic amino acids. The tendency of the rigid structure motif formation may have played a role in selecting for the homochirality that dominates life on Earth today.
Clinical significance of tumor-infiltrating lymphocytes and neutrophil-to-lymphocyte ratio in patients with stage III colon cancer who underwent surgery followed by FOLFOX chemotherapy
Authors: Cha Yoon Jin , Park Eun Jung , Baik Seung Hyuk , Lee Kang Young , Kang Jeonghyun
Published: August 2019

Local tumor immune response and host immunity have been suggested as important prognosticators respectively in colorectal cancer. However, the utility of combination of these parameters remains inconclusive. The aim of this study was to investigate the combinational impact of local and host tumor immune response, as determined by tumor-infiltrating lymphocytes (TILs) and neutrophil-to-lymphocyte ratio (NLR), in patients with stage III colon cancer. Patients with stage III colon cancer homogeneously treated with surgery followed by FOLFOX chemotherapy between Jan 2007 and Aug 2013 were included retrospectively. Hematoxylin and eosin (H&E) stained tumor sections of local inflammatory infiltrate (TILs) were classified as 0–3 by the Klintrup-Mäkinen grading method. NLR was measured within 1 month of surgery. The association of NLR and TILs with survival, alone or combined, were measured using multivariate Cox proportional hazard regression analysis. Among 137 patients, 75 (54.7%) were identified as the high TIL group (TILs 2 and 3) and 97 (70.8%) as the low NLR group (NLR < 3). Of the patients with high TILs, 51 (68%) had a low NLR. In univariate analysis, operation time, complications, lymph node ratio (LNR), stage, TILs, and high TILs with low NLR were significantly associated with overall survival(OS). Multivariate Cox regression identified operation time, stage, and TILs as independent risk factors for OS. When high TILs with low NLR vs. others was entered into multivariate analysis, this also proved to be a significant predictor of OS (HR 4.1, 95% CI 1.1–14.2, P = 0.025), with an increased C-index and lower AIC value compared to TILs. Measuring TILs using H&E stained sections could stratify the prognosis of stage III colon cancer. Considering host immunity, using the combination of TILs and NLR, allowed the prognosis to be stratified in more detail.
Neurotoxic potential of reactive astrocytes in canine distemper demyelinating leukoencephalitis
Authors: Klemens J. , Ciurkiewicz M. , Chludzinski E. , Iseringhausen M. , Klotz D. , Pfankuche V. M. , Ulrich R. , Herder V. , Puff C. , Baumgärtner W. , Beineke A.
Published: August 2019

Canine distemper virus (CDV) causes a fatal demyelinating leukoencephalitis in young dogs resembling human multiple sclerosis. Astrocytes are the main cellular target of CDV and undergo reactive changes already in pre-demyelinating brain lesions. Based on their broad range of beneficial and detrimental effects in the injured brain reactive astrogliosis is in need of intensive investigation. The aim of the study was to characterize astrocyte plasticity during the course of CDV-induced demyelinating leukoencephalitis by the aid of immunohistochemistry, immunofluorescence and gene expression analysis. Immunohistochemistry revealed the presence of reactive glial fibrillary acidic protein (GFAP)+ astrocytes with increased survivin and reduced aquaporin 4, and glutamine synthetase protein levels, indicating disturbed blood brain barrier function, glutamate homeostasis and astrocyte maladaptation, respectively. Gene expression analysis revealed 81 differentially expressed astrocyte-related genes with a dominance of genes associated with neurotoxic A1-polarized astrocytes. Accordingly, acyl-coA synthetase long-chain family member 5+/GFAP+, and serglycin+/GFAP+ cells, characteristic of A1-astrocytes, were found in demyelinating lesions by immunofluorescence. In addition, gene expression revealed a dysregulation of astrocytic function including disturbed glutamate homeostasis and altered immune function. Observed findings indicate an astrocyte polarization towards a neurotoxic phenotype likely contributing to lesion initiation and progression in canine distemper leukoencephalitis.
Enlarged Egg Size Increases Offspring Fitness of a Frog Species on the Zhoushan Archipelago of China
Authors: Xu Feng , Yang Weikang , Li Yiming
Published: August 2019

Egg size represents maternal investment and is an important life-history trait. It also is assumed to have an effect on offspring fitness. Life-history theory predicts that oviparous animals on islands will produce enlarged eggs because of increased maternal investment to improve offspring performance to facilitate intra-specific competition. The life-history theory, developed during the 1950s, provides a possible explanation for the “island rule”, but this rule has seldom been tested. Although several studies have detected a positive relationship between egg size and offspring fitness, it is difficult to exclude the covarying effects on offspring performance, such as genetic variation and developmental plasticity; predictions made using the life-history theory on the islands have not been tested. In this study, we have evaluated the relationship between egg size and offspring fitness on 20 islands in the Zhoushan Archipelago and two nearby mainland sites. To exclude covarying effects, we compared larval performance among different egg sizes in three levels: among siblings within clutches, among clutches within populations, and among different islands. The results showed that frogs on most of the islands did produce enlarged eggs and that their larvae had improved larval fitness. Additionally, at all three levels, the offspring that evolved from enlarged eggs had increased offspring fitness. The results of this study indicate that, for the first time, the life-history theory predictions concerning egg size and offspring fitness are supported.
Estimating nutrient uptake requirements for radish in China based on QUEFTS model
Authors: Zhang Jiajia , He Ping , Ding Wencheng , Xu Xinpeng , Ullah Sami , Abbas Tanveer , Ai Chao , Li Mingyue , Cui Rongzong , Jin Chongwei , Zhou Wei
Published: August 2019

Imbalanced fertilization has caused lower yield and nutrient use efficiency for radish (Raphanus sativus L.) production in China. Estimating nutrient requirements for radish is crucial in optimizing fertilization to resolve the problem. On-farm experiments in the radish-growing regions of China from 2000 to 2017 were collected to investigate the relationship between fleshy root yield and nutrient accumulation in radish plant using the Quantitative Evaluation of the Fertility of Tropical Soils (QUEFTS) model. The QUEFTS model predicted a linear increase in fleshy root yield if nutrients were taken up in balanced amounts until yield reached about 60%–70% of the potential yield. The balanced N, P, and K requirements in radish plant simulated by the QUEFTS model were 2.15, 0.45, and 2.58 kg to produce 1000 kg of fleshy root, and the corresponding internal efficiencies (IEs, kg fleshy root per kg nutrient in total plant dry matter) for N, P, and K were 465.1, 2222.2, and 387.1 kg kg−1. The simulated balanced N, P, and K removal by fleshy root to produce 1000 kg fleshy root were 1.34, 0.30, and 1.93 kg, respectively. Approximately 62%, 67%, and 75% of N, P, and K in radish plant were presented in the fleshy root and removed from the soil. Field validation experiments confirmed the consistency between the observed and simulated nutrient uptake values. The QUEFTS model was proven to be effective for estimating nutrient requirements of radish and will contribute to develop fertilizer recommendations for radish cultivated in China.
Dietary differences in archosaur and lepidosaur reptiles revealed by dental microwear textural analysis
Authors: Bestwick Jordan , Unwin David M. , Purnell Mark A.
Published: August 2019

Reptiles are key components of modern ecosystems, yet for many species detailed characterisations of their diets are lacking. Data currently used in dietary reconstructions are limited either to the last few meals or to proxy records of average diet over temporal scales of months to years, providing only coarse indications of trophic level(s). Proxies that record information over weeks to months would allow more accurate reconstructions of reptile diets and better predictions of how ecosystems might respond to global change drivers. Here, we apply dental microwear textural analysis (DMTA) to dietary guilds encompassing both archosaurian and lepidosaurian reptiles, demonstrating its value as a tool for characterising diets over temporal scales of weeks to months. DMTA, involving analysis of the three-dimensional, sub-micrometre scale textures created on tooth surfaces by interactions with food, reveals that the teeth of reptiles with diets dominated by invertebrates, particularly invertebrates with hard exoskeletons (e.g. beetles and snails), exhibit rougher microwear textures than reptiles with vertebrate-dominated diets. Teeth of fish-feeding reptiles exhibit the smoothest textures of all guilds. These results demonstrate the efficacy of DMTA as a dietary proxy in taxa from across the phylogenetic range of extant reptiles. This method is applicable to extant taxa (living or museum specimens) and extinct reptiles, providing new insights into past, present and future ecosystems.
Nature-Guided Synthesis of Advanced Bio-Lubricants
Authors: Romsdahl Trevor , Shirani Asghar , Minto Robert E. , Zhang Chunyu , Cahoon Edgar B. , Chapman Kent D. , Berman Diana
Published: August 2019

Design of environmentally friendly lubricants derived from renewable resources is highly desirable for many practical applications. Here, Orychophragmus violaceus (Ov) seed oil is found to have superior lubrication properties, and this is based on the unusual structural features of the major lipid species—triacylglycerol (TAG) estolides. Ov TAG estolides contain two non-hydroxylated, glycerol-bound fatty acids (FAs) and one dihydroxylated FA with an estolide branch. Estolide branch chains vary in composition and length, leading to their thermal stability and functional properties. Using this concept, nature-guided estolides of castor oil were synthesized. As predicted, they showed improved lubrication properties similar to Ov seed oil. Our results demonstrate a structure-based design of novel lubricants inspired by natural materials.