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Supplementary Table 1 List of antibodies, reagents and kits used in the study Supplementary Table 2 List of primers used in the study Supplementary Table 3 Comprehensive literature search of angiogenic regulators in HCC Supplementary Table 4 Correlation of 178 genes related to angiogenesis in HCC with SULF2 expression in human HCC microarray gene analysis

Existing antiangiogenic approaches to treat metastatic hepatocellular carcinoma (HCC) are weakly effectual, prompting further study of tumor angiogenesis in this disease setting. Here, we report a novel role for sulfatase 2 (SULF2) in driving HCC angiogenesis. Sulf2-deficient mice (Sulf2 KO) exhibited resistance to diethylnitrosamine-induced HCC and did not develop metastases like wild-type mice (Sulf2 WT). The smaller and less numerous tumors formed in Sulf2 KO mice exhibited a markedly lower microvascular density. In human HCC cells, SULF2 overexpression increased endothelial proliferation, adhesion, chemotaxis, and tube formation in a paracrine fashion. Mechanistic analyses identified the extracellular matrix protein periostin (POSTN), a ligand of αvβ3/5 integrins, as an effector protein in SULF2-induced angiogenesis. POSTN silencing in HCC cells attenuated SULF2-induced angiogenesis and tumor growth in vivo. The TGFβ1/SMAD pathway was identified as a critical signaling axis between SULF2 and upregulation of POSTN transcription. In clinical HCC specimens, elevated levels of SULF2 correlated with increased microvascular density, POSTN levels, and relatively poorer patient survival. Together, our findings define an important axis controlling angiogenesis in HCC and a mechanistic foundation for rational drug development. Cancer Res; 77(3); 632–45. ©2016 AACR.

Supplementary Figure 1 Schematic representation of experimental design for HCC carcinogenesis in mice. Supplementary Figure 2 Loss of Sulf2 impairs tumor growth and incidence in DEN-model of HCC carcinogenesis. Supplementary Figure 3 Conditioned media from SULF2-transfected HCC cells promotes endothelial cells viability and adhesion. Supplementary Figure 4 In silico analysis of HCC-angiogenic factors. Supplementary Figure 5 Expression of POSTN in Sulf2 knockout or wild type animals.

Citrinin (CTN), a secondary fungal metabolite produced by several Aspergillus, Penicillium, and Monascus genera species, is a toxin with a wide range of biological activities. Neutrophil extracellular traps represent a novel potential mechanism of the neutrophil response to foreign matters, and chicken heterophils can release similar heterophil extracellular traps (HETs). In this study, we aimed to investigate the effect of CTN on HET formation. Density gradient centrifugation was used to isolate chicken peripheral blood heterophils, and then immunofluorescence was used to observe the effects of CTN on HET formation. The mechanisms of HET formation were analyzed using pharmacological inhibitors and quantification of extracellular DNA, and the production of reactive oxygen species was detected with a fluorescent probe. Our results revealed that CTN (50-400 μM) had no cytotoxic effect on heterophils. CTN exposure induced the release of HETs composed of chromatin decorated with histones and elastase, and CTN-triggered HETs were dose- and time-dependent to some extent. Furthermore, CTN increased ROS production and activated p38 and ERK1/2 signaling pathways in heterophils. However, inhibition of the p38 signaling pathway, ERK1/2 signaling pathway, and NADPH oxidase pathway did not block HET formation induced by CTN. Inhibition of peptidyl arginine deiminase 4 (PAD4) enzyme and P2×1 receptor decreased HET formation after CTN stimulation, suggesting that HET formation exposed to CTN was mediated by PAD4 and P2×1 receptor. In conclusion, these findings may suggest a canonical mechanism relevant to the innate immunity caused by mycotoxins in chickens.

Acute liver injury is a severe clinical syndrome with markedly high mortality and poor prognosis. An accumulating body of evidence has demonstrated that epigenetic mechanisms have essential roles in the pathogenesis of acute liver injury. Lysine-specific demethylase 1 (LSD1) belongs to the amine oxidase superfamily of flavin adenine dinucleotide (FAD)-dependent enzymes, specifically demethylates H3 lysine 4. In the study, we investigated the effects and mechanisms of LSD1 in lipopolysaccharide (LPS)/D-Galactosamine (D-Gal)-induced acute liver injury in mice. Western blot analysis showed that LSD1 phosphorylation and di-methylated histone H3 on lysine 4 (H3K4me2) protein expression were significantly increased after LPS/D-Gal treatment (2.3 and 2.4 times higher than control respectively). GSK-LSD1 2HCl is an irreversible and selective LSD1 inhibitor. Pre-treatment with LSD1 inhibitor alleviated LPS/D-Gal-induced liver damage, decreased serum levels of alanine transaminase and aspartate aminotransferase in mice. Moreover, the LSD1 phosphorylation level in low, medium, and high LSD1 inhibitor groups was lower by a factor of 1.6, 1.9, and 2.0 from the LPS/D-Gal group, respectively. Mechanistically, LSD1 inhibitor further inhibited NF-κB signaling cascades and subsequently inhibited the production of pro-inflammatory cytokine TNF-α, IL-6, and IL-1β induced by LPS/D-Gal in liver tissues. Furthermore, LSD1 inhibitor upregulated the protein expression of Nrf2/HO-1 signaling pathways, and the activities of related antioxidant enzymes were enhanced. Collectively, our data demonstrated that LSD1 inhibitor protected against the LPS/D-Gal-induced acute liver injury via inhibiting inflammation and oxidative stress, and targeting the epigenetic marker may be a potent therapeutic strategy for acute liver injury.

Physicochemical characteristics, antioxidant activities and emulsifying properties of porcine plasma protein hydrolysates (PPPH) modified with oxidized tannic acid (OTA) or oxidized chlorogenic acid (OCA) were investigated. The formation of complexes was showed by increased turbidity, and both OTA and OCA were covalently attached to PPPH as shown by the significantly decrease in free amino group contents ( P P

The stability of rapeseed oil-in-water emulsions prepared with porcine plasma protein hydrolysates (PPPH) created via limited-hydrolysis was investigated with respect to concentrations (0.1, 0.5, 1.0, and 1.5%, w/w based on PPPH) of oxidized chlorogenic acid (OCA) added after emulsification. Compared to the emulsion with an absence of OCA, the incorporation of OCA induced significantly higher absolute ζ-potentials, smaller particle sizes (d3,2 and d4,3), lower flocculation factors (Ff) and lower coalescence indices (Ci) (P

Background: Recent studies showed that sushi repeat containing protein X linked 2 (SRPX2) could participate in the development of various malignant tumors. However, its role in non-small cell lung cancer (NSCLC) was unknown. The aim of the study was to prospectively investigate the role of SRPX2 in NSCLC cell proliferation, migration and invasion and reveal the underlying mechanism. Material and methods: Quantitative real-time polymerase chain reaction (qRT-PCR), western blot and immunohistochemistry – IHC) were used to measure detect the mRNA and protein levels, respectively, in NSCLC tissues and cell lines. Cell Counting Kit-8 (CCK-8), colony formation, wound healing and transwell assays were utilized to assess cell proliferation, migration and invasion. In vivo subcutaneous xenograft tumor model was established to detect the tumorigenic function of SRPX2, and IHC assay was performed to measure protein expression. Results: SRPX2 expression was upregulated in NSCLC tissues and cell lines, and positively correlated with tumor size, lymph node metastasis, distant metastasis and clinical stage. High SRPX2 expression also predicted poor prognosis. In vitro experiments indicated that overexpression of SRPX2 promoted the proliferation, migration, and invasion of SPC-A1 cells while knockdown of SRPX2 caused the opposite effects in A549 cells. Specifically, SRPX2 activated FAK/SRC/ERK pathway and its downstream effectors and promoted epithelial-mesenchymal transition (EMT). Conclusion: Taken together, our findings revealed a functional role of SRPX2 in NSCLC cell proliferation, migration and invasion. The underlying mechanism was, at least partially, the activation of FAK/SRC/ERK pathway. This study provides the molecular basis for targeting SRPX2 in potential clinical application for NSCLC.

Recent studies showed that sushi repeat containing protein X linked 2 (SRPX2) could participate in the development of various malignant tumors. However, its role in non-small cell lung cancer (NSCLC) was unknown. The aim of the study was to prospectively investigate the role of SRPX2 in NSCLC cell proliferation, migration and invasion and reveal the underlying mechanism.Quantitative real-time polymerase chain reaction (qRT-PCR), western blot and immunohistochemistry - IHC) were used to measure detect the mRNA and protein levels, respectively, in NSCLC tissues and cell lines. Cell Counting Kit-8 (CCK-8), colony formation, wound healing and transwell assays were utilized to assess cell proliferation, migration and invasion. In vivo subcutaneous xenograft tumor model was established to detect the tumorigenic function of SRPX2, and IHC assay was performed to measure protein expression.SRPX2 expression was upregulated in NSCLC tissues and cell lines, and positively correlated with tumor size, lymph node metastasis, distant metastasis and clinical stage. High SRPX2 expression also predicted poor prognosis. In vitro experiments indicated that overexpression of SRPX2 promoted the proliferation, migration, and invasion of SPC-A1 cells while knockdown of SRPX2 caused the opposite effects in A549 cells. Specifically, SRPX2 activated FAK/SRC/ERK pathway and its downstream effectors and promoted epithelial-mesenchymal transition (EMT).Taken together, our findings revealed a functional role of SRPX2 in NSCLC cell proliferation, migration and invasion. The underlying mechanism was, at least partially, the activation of FAK/SRC/ERK pathway. This study provides the molecular basis for targeting SRPX2 in potential clinical application for NSCLC.

This study investigated the effect of different concentrations of porcine plasma protein hydrolysates (PPPH) on the gelation of porcine myofibrillar protein (MP) exposed to a hydroxyl radical-generating system (HRGS). Compared to non-oxidized MP, the oxidative modification led to a decreased penetration force and water holding capacity (WHC) ( P 0.05). Meanwhile, addition of PPPH (1.5 mg/mL) reduced the loss of MP gel strength and WHC induced by HRGS ( P 0.05). Low-field nuclear magnetic resonance results suggested that the addition of PPPH facilitated the hydration of the protein. The results concerning molecular forces revealed that hydrophobic interactions, hydrogen bonds, and disulphide bonds are the primary forces in gel formation. Non-reducing electrophoresis indicated that the addition of PPPH reduced protein loss and aggregation. The addition of PPPH promoted the formation of a more smooth and homogeneous gel network. Our results demonstrated that PPPH effectively retarded oxidation-induced MP gel deterioration and could be used as an antioxidant in meat products.

In this work, a oxalamide derivative, N 1 ,N 1′ -(ethane-1,2-diyl)bis( N 2 -phenyloxalamide) (EDPO) was synthesized successfully to prepare the poly(1,4-butylene adipate) PBA/EDPO composite. The EDPO first crystallized into the rod-like (shish) and induced the PBA crystal aligned perpendicularly to the EDPO rod axis. As an effective nucleating agent, the EDPO highly increased the crystallization rate and shortened the crystallization time of the PBA. The EDPO not only facilitated the formation of the PBA α -crystal and modulated the polymorphic crystalline structure of the PBA, but also accelerated the β -to- α phase transition of the PBA. Upon the extremely low (such as 0.1%) weight fraction of the EDPO, the thermal stability of the PBA increased pronouncedly. The mechanisms on the nucleation and growth of the PBA, the preferential formation of the α -crystal, accelerated phase transition and enhanced thermal stability of the PBA were also proposed and discussed.

In this work, the decamethylenedicarboxylic dibenzoylhydrazide (DMC-DBH) is attempted as a nucleating agent (NA) to introduce into the poly(1,4-butylene adipate) (PBA) matrix. Effects of the DMC-DBH on the PBA, including the crystallization kinetics, crystal morphology, polymorphic crystalline structure, and biodegradation behaviors, are investigated by differential scanning calorimetry (DSC), polarized optical microscopy (POM), wide-angle X-ray diffraction (WAXD), Fourier transform infrared (FTIR) spectroscopy, and scanning electron microscopy (SEM). The crystallization temperature and crystallization rate of the PBA increase upon incorporation of the DMC-DBH. This composite is characterized by a distinctive shish-kebab structure. The DMC-DBH is favorable for formation of the PBA α-crystal at markedly lower melting-crystallization temperature (Tc), compared to the neat PBA and accelerated β-to-α phase transition. The biodegradation rate of the PBA decreased upon addition of the DMC-DBH. The mechanisms on...

Zinc phenylphosphonate (PPZn), a benign and biocompatible nucleating agent, was prepared and incorporated into the biodegradable poly(ethylene adipate) (PEA) to investigate its effect on the crystallization behavior, crystallization kinetics and spherulite morphology of PEA. Upon addition of PPZn, the crystallization temperature and crystallinity of PEA in the non-isothermal crystallization process increased significantly. Analysis of crystallization kinetics by Avrami equation suggests that the crystallization time shortened greatly and crystallization rate increased markedly after addition of PPZn. In the presence of PPZn, the spherulite size decreased and spherulite density increased significantly. It suggests that PPZn is an efficient nucleating agent for the crystallization of PEA. The accelerated crystallization in the presence of PPZn is mainly attributed to the epitaxial nucleation of PEA crystals on the surface of PPZn crystals, that is, a perfect lattice matching between PEA crystal and PPZn crystal occurs.

Fibrosis is a common pathologic outcome of chronic disease resulting in the replacement of normal tissue parenchyma with a collagen-rich extracellular matrix produced by myofibroblasts. Although the progenitor cell types and cellular programs giving rise to myofibroblasts through mesenchymal transition can vary between tissues and diseases, their contribution to fibrosis initiation, maintenance, and progression is thought to be pervasive. Here we showed that the ability of transforming growth factor-β (TGFβ) to efficiently induce myofibroblast differentiation of cultured epithelial cells, endothelial cells, or quiescent fibroblasts is dependent upon the induced expression and activity of dimeric calpains, a family of non-lysosomal cysteine proteases that regulate a variety of cellular events through post-translational modification of diverse substrates. siRNA-based gene silencing demonstrated that TGFβ-induced mesenchymal transition of a murine breast epithelial cell line was dependent upon induction of expression of calpain 9 (CAPN9), an isoform previously thought to be restricted to the gastrointestinal tract. Mice lacking functional CAPN9 due to biallelic targeting of Capn9 were viable and fertile, but showed overt protection from bleomycin-induced lung fibrosis, carbon tetrachloride-induced liver fibrosis, and angiotensin II-induced cardiac fibrosis and dysfunction. A predicted loss-of-function allele of CAPN9 is common in Southeast Asia, with the frequency of homozygosity matching the prediction of Hardy-Weinberg equilibrium; together with the highly spatially-restricted pattern of CAPN9 expression under physiologic circumstances and the heartiness of the murine knockout, these data provide a strong signature for tolerance of therapeutic strategies for fibrosis aimed at CAPN9 antagonism.

The objective of this study was to evaluate the effect of two different forms of canola oil (pure liquid form or a pre-emulsified form that includes porcine plasma protein hydrolysates modified by oxidized tannic acid) used for pork back-fat substitutions on the physico-chemical characteristics of frankfurters. When compared to the control group, partial replacement of pork back-fat using a pre-emulsified canola oil system exhibited excellent water and fat binding capacities, quicker relaxation times and lower amounts of saturated fatty acids (SFAs) (P .05), as well as increased hardness, gumminess, and chewiness (P .05) when verified by dynamic rheology analysis. Additionally, higher replacement ratio of pre-emulsified canola oil did significantly decrease the overall acceptability than the control group (P .05). Our results indicate that pre-emulsified canola oil provided greater improvement with respect to the physical characteristics of partial pork back-fat replaced frankfurters, when compared to pure canola oil inclusion.

Layered metal phosphonate, zinc phenylphosphonate (PPZn) was synthesized and used as a model benign nucleating agent for poly(butylene adipate) (PBA) to manipulate the crystallization behavior, polymorphic crystalline structure and biodegradation of PBA. Upon incorporation of PPZn, the crystallization temperature (Tc) and crystallinity of PBA increased. Meanwhile, the crystallization rate and spherulite density of PBA increased in the presence of PPZn, indicating that PPZn has an effective nucleating effect for PBA. PPZn is favorable for the formation of the PBA α-modification. Furthermore, PPZn would speed up highly the phase transition of PBA from the β- to α-one. PPZn significantly suppressed the biodegradation of PBA. Mechanisms for nucleating, preferential formation of the α-modification, accelerated phase transition and decreased degradation rate of PBA in the presence of PPZn have also been proposed. It is considered that layered metal phosphonate material is a new type of benign filler for controlling the polymorphic crystalline structure and biodegradation rate of the polymer, which is expected to further modify the property of the polymer and might widen the applications of biodegradable materials in the environmental and biomedical fields.

Women with Marfan syndrome (MFS) are at high risk for pregnancy-associated aortic dissection. Pathogenic models that singularly invoke hemodynamic stress are difficult to reconcile with predominant postnatal occurrence of aortic tear, often occurring weeks to months after delivery. In consideration of events that peak at term, are sustained after delivery, and might synergize with previously defined signaling pathways implicated in aneurysm progression, we examined the hormone oxytocin, which initiates uterine contraction and milk letdown for the duration of lactation through phosphorylation of extracellular signal-regulated kinase (ERK). In a mouse model of MFS that shows highly penetrant postnatal aortic dissection, risk was strongly attenuated by preventing lactation or use of an oxytocin receptor antagonist. Survival correlated inversely with the extent of ERK activation in the aortic wall, and strong protection was observed upon attenuation of ERK phosphorylation using an inhibitor of ERK kinase (MEK) or the U.S. Food and Drug Administration-approved medication hydralazine, offering potential therapeutic strategies for pregnancy-associated vascular catastrophe in the setting of MFS.

Effects of cyanuric acid (CA) on the isothermal and nonisothermal crystallization, morphology, polymorphic crystalline structure, and phase transition on PBA have been investigated systematically by differential scanning calorimetry (DSC), polarized optical microscopy (POM), wide-angle X-ray diffraction (WAXD), and Fourier transform infrared (FTIR) spectroscopy. The nonisothermal crystallization temperature increased upon addition of CA. The crystallization kinetics of isothermal crystallization was quantitatively analyzed. CA is an effective nucleating agent for the enhanced nucleation density and decreased spherulite size of PBA. Addition of CA was favorable for the formation of PBA α modification and accelerated the phase transition from the β modification to the α modification. The mechanisms of nucleation, preferential formation of PBA α modification, and accelerated phase transition have also been proposed.

Existing antiangiogenic approaches to treat metastatic hepatocellular carcinoma (HCC) are weakly effectual, prompting further study of tumor angiogenesis in this disease setting. Here, we report a novel role for sulfatase 2 (SULF2) in driving HCC angiogenesis. Sulf2-deficient mice (Sulf2 KO) exhibited resistance to diethylnitrosamine-induced HCC and did not develop metastases like wild-type mice (Sulf2 WT). The smaller and less numerous tumors formed in Sulf2 KO mice exhibited a markedly lower microvascular density. In human HCC cells, SULF2 overexpression increased endothelial proliferation, adhesion, chemotaxis, and tube formation in a paracrine fashion. Mechanistic analyses identified the extracellular matrix protein periostin (POSTN), a ligand of avb3/5 integrins, as an effector protein in SULF2-induced angiogenesis. POSTN silencing in HCC cells attenuated SULF2-induced angiogenesis and tumor growth in vivo. The TGFb1/SMAD pathway was identified as a critical signaling axis between SULF2 and upregulation of POSTN transcription. In clinical HCC specimens, elevated levels of SULF2 correlated with increased microvascular density, POSTN levels, and relatively poorer patient survival. Together, our findings define an important axis controlling angiogenesis in HCC and a mechanistic foundation for rational drug development. Fil: Chen, Gang. Mayo Clinic; Estados Unidos. Wenzhou Medical University; China Fil: Nakamura, Ikuo. Mayo Clinic; Estados Unidos Fil: Dhanasekaran, Renumathy. Mayo Clinic; Estados Unidos. University of Stanford; Estados Unidos Fil: Iguchi, Eriko. Schulze Center For Novel Therapeutics, Mayo Clinic; Estados Unidos Fil: Tolosa, Ezequiel Julian. Schulze Center For Novel Therapeutics, Mayo Clinic; Estados Unidos. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; Argentina Fil: Romecin, Paola A.. Schulze Center For Novel Therapeutics, Mayo Clinic; Estados Unidos Fil: Vera, Renzo Emanuel. Schulze Center For Novel Therapeutics, Mayo Clinic; Estados Unidos. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Biología Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina Fil: Almada, Luciana Victoria. Schulze Center For Novel Therapeutics, Mayo Clinic; Estados Unidos Fil: Miamen, Alexander G.. Mayo Clinic; Estados Unidos Fil: Chaiteerakij, Roongruedee. Mayo Clinic; Estados Unidos Fil: Zhou, Mengtao. Wenzhou Medical University; China Fil: Asiedu, Michael K.. Mayo Clinic; Estados Unidos Fil: Moser, Catherine D.. Mayo Clinic; Estados Unidos Fil: Han, Shaoshan. Mayo Clinic; Estados Unidos Fil: Hu, Chunling. Mayo Clinic; Estados Unidos Fil: Banini, Bubu A.. Mayo Clinic; Estados Unidos Fil: Oseini, Abdul M.. Mayo Clinic; Estados Unidos Fil: Chen, Yichun. Mayo Clinic; Estados Unidos Fil: Fang, Yong. Mayo Clinic; Estados Unidos Fil: Yang, Dongye. Mayo Clinic; Estados Unidos Fil: Shaleh, Hassan M.. Mayo Clinic; Estados Unidos Fil: Wang, Shaoqing. Mayo Clinic; Estados Unidos Fil: Wu, Dehai. Mayo Clinic; Estados Unidos Fil: Song, Tao. Mayo Clinic; Estados Unidos Fil: Lee, Ju-Seog. Anderson Cancer Center; Estados Unidos Fil: Thorgeirsson, Snorri S.. National Cancer Institute; Estados Unidos Fil: Chevet, Eric. Université Rennes; Francia Fil: Shah, Vijay H.. Mayo Clinic; Estados Unidos Fil: Fernandez-Zapico, Martin E.. Schulze Center For Novel Therapeutics, Mayo Clinic; Estados Unidos Fil: Roberts, Lewis R.. Mayo Clinic; Estados Unidos

In this present study, strongly cationic coagulants were prepared by modifying chitosan with 3-chloro-2-hydroxypropyl trimethyl ammonium (CTA). The coagulation properties of this cationic chitosan (chitosan-CTA) were systematically examined in laboratory and pilot scale studies. At the laboratory scale, the effects of coagulant dosage, temperature, initial turbidity of untreated water, pH, and salinity were investigated by a jar test using kaolin suspension as synthetic water. Compared with several conventional coagulants, chitosan-CTA showed a lower dosage requirement of approximately 0.05 mg/L, but higher turbidity removal efficiency of approximately 93.3%. Moreover, raw water from the Zhenjiang part of the Yangtze River in China was used as test water for further investigation on a pilot scale. The influences of dosage, sedimentation time, and mechanical mixing rate on the coagulation properties were studied by an orthogonal test and the optimal experimental conditions were obtained. The dosag...

In this study, a kind of amphoteric chitosan-based flocculants (3-chloro-2-hydroxypropyl trimethyl ammonium chloride (CTA) modified carboxymethyl chitosan, denoted as CMC-CTA) have been prepared. The amphoteric flocculants showed notable improvement in solubility and salt-resistance in the whole pH range compared with its precursors, i.e., chitosan and cationic chitosan. The flocculation properties of CMC-CTA were systematically studied at both the laboratory and pilot scales. At laboratory scale, kaolin suspension was employed as synthetic water in the beaker experiment. The effects of pH, temperature, and original turbidity of untreated water on the dosage and flocculation performances were investigated, respectively. At pH below and above the isoelectric point of CMC-CTA, the optimal flocculation performances were obtained with a dosage of 0.03–0.20 mg/L and 0.8–1.0 mg/L, respectively. The flocculation mechanism was discussed in detail from the morphologies of polymeric flocculants and flocculation kinetics: The shape and charge of polymer chains in the solution greatly influenced the final flocculation performances; The flocculation kinetics agreed well with the particle collision model. Moreover, raw water from the Zhenjiang part of the Yangtze River in China was used for the further investigation of the flocculation properties at the pilot scale. The influences of dosage, sedimentation time, and mechanical mixing rate to the flocculation properties were studied by orthogonal testing. The optimal experimental condition was then obtained. Compared with the currently used flocculants in the Zhenjiang Water Factory, i.e., polyferric sulfate, which has an optimal dosage around 40 mg/L, the value of CMC-CTA (1.1 mg/L) decreased greatly. Above all, the flocculation experiments at the laboratory and pilot scales both indicate that the amphoteric chitosan-based flocculants have good flocculation performances in a wide pH range.

Transforming growth factor–β (TGFβ) signaling drives aneurysm progression in multiple disorders, including Marfan syndrome (MFS), and therapies that inhibit this signaling cascade are in clinical trials. TGFβ can stimulate multiple intracellular signaling pathways, but it is unclear which of these pathways drives aortic disease and, when inhibited, which result in disease amelioration. Here we show that extracellular signal–regulated kinase (ERK) 1 and 2 and Smad2 are activated in a mouse model of MFS, and both are inhibited by therapies directed against TGFβ. Whereas selective inhibition of ERK1/2 activation ameliorated aortic growth, Smad4 deficiency exacerbated aortic disease and caused premature death in MFS mice. Smad4-deficient MFS mice uniquely showed activation of Jun N-terminal kinase–1 (JNK1), and a JNK antagonist ameliorated aortic growth in MFS mice that lacked or retained full Smad4 expression. Thus, noncanonical (Smad-independent) TGFβ signaling is a prominent driver of aortic disease in MFS mice, and inhibition of the ERK1/2 or JNK1 pathways is a potential therapeutic strategy for the disease.

The effect of anhydrous orotic acid (OA), as a biocompatible nucleating agent (NA), on the non-isothermal and isother-mal crystallization behaviors, polymorphic crystalline structure and phase transition of poly(1,4-butylene adipate) (PBA) was investi-gated. It is found that the OA increased the crystallization temperature of the PBA in the non-isothermal crystallization process anddecreased the crystallization time of the PBA in the isothermal crystallization process. Meanwhile, the spherulite size decreased andspherulite density increased for the PBA. The OA favored the formation of the PBA a-form crystal, compared to the neat PBA. Inaddition, upon incorporation of the OA, the b-to-a phase transition rate was enhanced significantly. Mechanisms for the preferentialformation of the PBA a-form crystal and the accelerated phase transition have also been proposed. V C 2015 Wiley Periodicals, Inc. J. Appl.Polym. Sci. 2016, 133, 42957. KEYWORDS: biodegradable; composites; crystallization; polyesters; properties and characterizationReceived 31 July 2015; accepted 21 September 2015DOI: 10.1002/app.42957INTRODUCTION

Kabuki syndrome is caused by haploinsufficiency for either of two genes that promote the opening of chromatin. If an imbalance between open and closed chromatin is central to the pathogenesis of Kabuki syndrome, agents that promote chromatin opening might have therapeutic potential. We have characterized a mouse model of Kabuki syndrome with a heterozygous deletion in the gene encoding the lysine-specific methyltransferase 2D (Kmt2d), leading to impairment of methyltransferase function. In vitro reporter alleles demonstrated a reduction in histone 4 acetylation and histone 3 lysine 4 trimethylation (H3K4me3) activity in mouse embryonic fibroblasts from Kmt2d(+/βGeo) mice. These activities were normalized in response to AR-42, a histone deacetylase inhibitor. In vivo, deficiency of H3K4me3 in the dentate gyrus granule cell layer of Kmt2d(+/βGeo) mice correlated with reduced neurogenesis and hippocampal memory defects. These abnormalities improved upon postnatal treatment with AR-42. Our work suggests that a reversible deficiency in postnatal neurogenesis underlies intellectual disability in Kabuki syndrome.

Loeys-Dietz syndrome (LDS) is a connective tissue disorder that is characterized by a high risk for aneurysm and dissection throughout the arterial tree and phenotypically resembles Marfan syndrome. LDS is caused by heterozygous missense mutations in either TGF-β receptor gene (TGFBR1 or TGFBR2), which are predicted to result in diminished TGF-β signaling; however, aortic surgical samples from patients show evidence of paradoxically increased TGF-β signaling. We generated 2 knockin mouse strains with LDS mutations in either Tgfbr1 or Tgfbr2 and a transgenic mouse overexpressing mutant Tgfbr2. Knockin and transgenic mice, but not haploinsufficient animals, recapitulated the LDS phenotype. While heterozygous mutant cells had diminished signaling in response to exogenous TGF-β in vitro, they maintained normal levels of Smad2 phosphorylation under steady-state culture conditions, suggesting a chronic compensation. Analysis of TGF-β signaling in the aortic wall in vivo revealed progressive upregulation of Smad2 phosphorylation and TGF-β target gene output, which paralleled worsening of aneurysm pathology and coincided with upregulation of TGF-β1 ligand expression. Importantly, suppression of Smad2 phosphorylation and TGF-β1 expression correlated with the therapeutic efficacy of the angiotensin II type 1 receptor antagonist losartan. Together, these data suggest that increased TGF-β signaling contributes to postnatal aneurysm progression in LDS.

A series of biodegradable amphoteric chitosan-based flocculants (3-chloro-2-hydroxypropyl trimethyl ammonium chloride (CTA) modified carboxymethyl chitosan, denoted as CMC-CTA) with different substitution degrees of CTA were prepared successfully. The content of carboxymethyl groups in each CMC-CTA sample was kept almost constant. The solubility of the various flocculants showed that, higher cationic content of flocculants caused a better solubility. The flocculation experiments using kaolin suspension as synthetic water at the laboratory scale indicated that the substitution degree of CTA was one of the key factors for the flocculation properties. With the increase of cationic content, the flocculants were demonstrated better flocculation performance and lower dosage requirement. Flocculation kinetics model of particles collisions combining zeta potential and turbidity measurements was employed to investigate the effects of the cationic content of the flocculants on the flocculation properties from the viewpoint of flocculation mechanism in detail. Furthermore, flocculation performance using raw water from Zhenjiang part of Yangtze River at the pilot scale showed the similar effects to those at the laboratory scale.

Item does not contain fulltext Loeys-Dietz syndrome (LDS) associates with a tissue signature for high transforming growth factor (TGF)-beta signaling but is often caused by heterozygous mutations in genes encoding positive effectors of TGF-beta signaling, including either subunit of the TGF-beta receptor or SMAD3, thereby engendering controversy regarding the mechanism of disease. Here, we report heterozygous mutations or deletions in the gene encoding the TGF-beta2 ligand for a phenotype within the LDS spectrum and show upregulation of TGF-beta signaling in aortic tissue from affected individuals. Furthermore, haploinsufficient Tgfb2(+/-) mice have aortic root aneurysm and biochemical evidence of increased canonical and noncanonical TGF-beta signaling. Mice that harbor both a mutant Marfan syndrome (MFS) allele (Fbn1(C1039G/+)) and Tgfb2 haploinsufficiency show increased TGF-beta signaling and phenotypic worsening in association with normalization of TGF-beta2 expression and high expression of TGF-beta1. Taken together, these data support the hypothesis that compensatory autocrine and/or paracrine events contribute to the pathogenesis of TGF-beta-mediated vasculopathies.

A functional design diagram for one of parallel hybrid electric vehicles performance simulation software was presented. Using the Unified Modeling Language modeling mechanism, the software system's architecture diagrams, using case diagrams and class diagrams were established. By giving an example for using the software, a compare was made between the software's result and another result which is concluded from ADVISOR 2002 simulation on the condition of same vehicle parameters. It is illustrating that the software is usable and accuracy.

Angiotensin II (AngII) mediates progression of aortic aneurysm, but the relative contribution of its type 1 (AT1) and type 2 (AT2) receptors remains unknown. We show that loss of AT2 expression accelerates the aberrant growth and rupture of the aorta in a mouse model of Marfan syndrome (MFS). The selective AT1 receptor blocker (ARB) losartan abrogated aneurysm progression in the mice; full protection required intact AT2 signaling. The angiotensin-converting enzyme inhibitor (ACEi) enalapril, which limits signaling through both receptors, was less effective. Both drugs attenuated canonical transforming growth factor–β (TGFβ) signaling in the aorta, but losartan uniquely inhibited TGFβ-mediated activation of extracellular signal–regulated kinase (ERK), by allowing continued signaling through AT2. These data highlight the protective nature of AT2 signaling and potentially inform the choice of therapies in MFS and related disorders.