Search

Biochar produced from bio-wastes has been widely used to promote the performance of anaerobic digestion. Waste activated sludge (WAS) is considered as a kind of popular precursor for biochar preparation, but the abundant resources in WAS were neglected previously. In this study, the roles of biochar prepared from raw, pretreated, and fermented sludge on anaerobic digestion were investigated. That is, parts of carbon sources and nutrients like polysaccharides, proteins, and phosphorus were firstly recovered after sludge pretreatment or fermentation, and then the sludge residuals were used as raw material to prepare biochar. The methane yield improved by 22.1% with adding the biochar (AK-BC) prepared by sludge residual obtained from alkaline pretreatment. Mechanism study suggested that the characteristics of AK-BC like specific surface area and defect levels were updated. Then, the conversion performance of intermediate metabolites and electro-activities of extracellular polymeric substances were up-regulated. As a result, the activity of electron transfer was increased with the presence of AK-BC, with increase ratio of 21.4%. In addition, the electroactive microorganisms like Anaerolineaceae and Methanosaeta were enriched with the presence of AK-BC, and the potential direct interspecies electron transfer was possibly established. Moreover, both aceticlastic and CO 2 -reducing methanogenesis pathways were improved by up-regulating related enzymes. Therefore, the proposed strategy can not only obtain preferred biochar but also recover abundant resources like carbon source, nutrients, and bioenergy. [Display omitted] • Biochar from raw, pretreated, and fermented sludge residuals were optimized. • Methane yield increased by 22.1% with biochar from alkaline pretreated sludge. • Alkaline pretreated sludge biochar increased electron transport activity by 21.4%. • Both aceticlastic and CO 2 -reducing methanogenesis pathways were improved. • A green and sustainable strategy for final disposal of sludge was proposed. [ABSTRACT FROM AUTHOR]

The tumor microenvironment and interplay with cancer cells could promote tumor growth and metastasis. Here we report that polarization state of macrophages could affect epithelial–mesenchymal transition (EMT) and mesenchymal–epithelial transition (MET). IL-35 level secreted by M1 macrophage was significantly higher than M2 macrophage and it facilitated EMT process through activation of STAT3 in hepatocellular carcinoma cells. Interestingly, IL-35 could not directly promote MET, but it could indirectly induce MET of HCC cells through M2 macrophage polarization. These results indicated the level of IL-35 in tumor microenvironment may fluctuate at different stages of oncogenesis to regulate epithelial plasticity of HCC and provide potential therapeutic targets for tumor metastasis. • IL-35 level secreted by M1 macrophage was significantly higher than M2 macrophage. • M1 secreted IL-35 was not sufficient to promote MET whereas it facilitated EMT through STAT3 activation in HCC cells. • IL-35 could not directly promote MET whereas it could indirectly induce MET of HCC cells through M2 polarization. [ABSTRACT FROM AUTHOR]

Elucidating the mechanism underlying osteoclast differentiation is important to improve our understanding of the pathophysiologies related to skeletal diseases and osteolytic metastasis in cancer. Sex-determining region Y-box containing gene 2 (SOX2), a stemness marker, is known to affect osteoblast differentiation and cancer metastasis. However, its role in osteoclastogenesis has not been investigated to date. Here, we report that SOX2 protein and mRNA expression was upregulated during osteoclast differentiation. The overexpression or knockdown of SOX2 in osteoclast precursor cells enhanced or suppressed, respectively, receptor activator of nuclear factor κB ligand (RANKL)-induced osteoclast differentiation and migration, and nuclear factor of activated T-cell c1 (NFATc1) and factor-associated suicide ligand (FASL) expression. In addition, epidermal growth factor receptor (EGFR) and extracellular signal-regulated kinase (ERK) activation were regulated by SOX2 expression; both EGFR and ERK inhibitors abrogated the SOX2 overexpression-induced increase in osteoclast differentiation and NFATc1 expression under RANKL stimulation. Overall, these results suggest SOX2 as a positive regulatory factor during osteoclast differentiation partly through the EGFR and ERK signaling pathways, highlighting a new potential target for restoring abnormal osteoclast activation. • The effects and role of SOX2 in osteoclastogenesis were investigated. • SOX2 positively regulates RANKL-induced osteoclast differentiation. • The regulation mechanism involves modulating NFATc1 expression and EGFR/ERK signaling. • SOX2 is a promising candidate for treatment of abnormal osteoclast activation. [ABSTRACT FROM AUTHOR]

Despite its rich content, iron bioavailability from leafy greens is low relative to protoporphyrin-bound (e.g. heme) iron, richest in red meat. We hypothesized that combinations of novel food processing techniques, i.e., vacuum impregnation (VI), water-bath (WB), overnight immersion (IM) in an iron sulfate solution, and moderate electric field (MEF) could improve kale leaf iron content and produce protoporphyrin-bound iron derivatives in situ. Combination MEF treatments increased leaf iron concentrations ∼160x greater than control. A ferroprotoporphyrin IX isomer was detected in MEF and IM-treated kale, providing up to 0.75 mg protoporphyrin-bound iron/100 g kale leaf fresh weight (FW). IM and MEF treatments also significantly increased leaf total chlorophyll concentrations ∼3x greater than the control. The novel food processing techniques employed herein produced kale with significantly higher concentrations of free + heme-bound iron, as well as total chlorophyll, relative to other plant-based sources. Future testing will determine if this "iron-enhanced" kale delivers more iron, as compared to other iron-rich food sources. • Combination MEF treatments increased leaf total iron concentrations up to ∼160× more than control. • Combination MEF treatments also increased kale total chlorophyll derivatives by ∼3×. • Protoporphyrin-bound iron was identified and quantitated in combination-treated kale leaves. [ABSTRACT FROM AUTHOR]

Twelve novel fenfuram-diarylether hybrids were designed, synthesized and characterized by 1 H NMR and MS. Their in vitro antifungal activities were evaluated against five phytopathogenic fungi by mycelial growth inhibition method. Most compounds showed significant antifungal effect on Rhizoctonia solani and Sclerotinia sclerotiorum . Compound 1c exhibited the most potent antifungal effect on R. solani with an EC 50 value of 0.242 mg/L, superior to the commercial fungicide boscalid (EC 50 = 1.758 mg/L) and the lead fungicide fenfuram (EC 50 = 7.691 mg/L). Molecular docking revealed that compound 1c featured a higher affinity for succinate dehydrogenase (SDH) than fenfuram. Furthermore, it was shown that the 2-chlorophenyl group of compound 1c formed a π-π stacking with D/Tyr-128 and a Cl-π interaction with B/His-249, which made compound 1c more active than fenfuram against SDH. [ABSTRACT FROM AUTHOR]

In order to exploit their industrial applications, sericin peptides (3K-SP) were obtained by membrane separation and their physiochemical properties and cryoprotective function were investigated. Results showed that 3K-SP were mostly distributed less than 3000 Da, and rich in the amino acids Ser, Asp, Gly, Thr and Glu, which have been associated with the cryoprotective activity of ice-structuring proteins. Addition of 3K-SP to a frozen solution led to the reductions in melting temperature and melting time compared to control solution. In addition, 3K-SP inhibited ice recrystallization, since it could maintain small ice crystal sizes within a frozen solution. Furthermore, 3K-SP demonstrated high cryogenic protection activity of Lactobacillus delbrueckii Subsp. Bulgaricus during freezing, and provided optimal protection of cells at conditions in which its concentration was 1.0 mg/mL and the pH of the solution was 7.0. In these conditions, the percentage of surviving cells was as high as 84.78 ± 3.07%. Flow cytometric and scanning electron microscopy analyses showed that treatment with 3K-SP increased the percentage of viable cells from 52.9% to 80.1%, and suggest that 3K-SP may mediate its protective effects through interaction with cell membranes, whereby it surrounds cells in a glassy matrix that helps maintain their membrane integrity. [ABSTRACT FROM AUTHOR]

[Display omitted] • The phylogeny of Asian Sonerileae was reconstructed using genome resequencing data. • Thirty-four major clades were identified in Asian Sonerileae. • Widespread phylogenetic discordance was caused by a combination of factors. • We detected high level of homoplasy for some characters used in taxonomy. • The backbone phylogeny of Asian Sonerileae offers a solid basis for future revision. Sonerileae is a diverse Melastomataceae lineage comprising ca. 1000 species in 44 genera, with >70% of genera and species distributed in Asia. Asian Sonerileae are taxonomically intractable with obscure generic circumscriptions. The backbone phylogeny of this group remains poorly resolved, possibly due to complexity caused by rapid species radiation in early and middle Miocene, which hampers further systematic study. Here, we used genome resequencing data to reconstruct the phylogeny of Asian Sonerileae. Three parallel datasets, viz. single-copy ortholog (SCO), genomic SNPs, and whole plastome, were assembled from genome resequencing data of 205 species for this purpose. Based on these genome-scale data, we provided the first well resolved phylogeny of Asian Sonerileae, with 34 major clades identified and 74% of the interclade relationships consistently resolved by both SCO and genomic data. Meanwhile, widespread phylogenetic discordance was detected among SCO gene trees as well as species trees reconstructed using different tree estimation methods (concatenation/site-based coalescent method/summary method) or different datasets (SCO/genomic/plastome). We explored sources of discordance using multiple approaches and found that the observed discordance in Asian Sonerileae was mainly caused by a combination of biased distribution of missing data, random noise from uninformative genes, incomplete lineage sorting, and hybridization/introgression. Exploration of these sources can enable us to generate hypotheses for future testing, which is the first step towards understanding the evolution of Asian Sonerileae. We also detected high levels of homoplasy for some characters traditionally used in taxonomy, which explains current chaotic generic delimitations. The backbone phylogeny of Asian Sonerileae revealed in this study offers a solid basis for future taxonomic revision at the generic level. [ABSTRACT FROM AUTHOR]

The aryl hydrocarbon receptor (AHR) is a ligand-activated transcription factor with pleiotropic effects in normal physiology or vascular development, xenobiotic metabolism, and cancer. A previous study has reported that BRG1, a component of the SWI/SNF complex, is a coactivator for AHR and is recruited to the promoter region of the CYP1A1 gene in mouse hepatocytes. Recent data suggest that AHR is also expressed in human retinal pigment epithelial cells (ARPE-19), which play a crucial role in retinal physiology and the visual cycle. Multiple studies have shown that the AHR plays an important role in the pathogenesis of retinal diseases including age-related macular degeneration. However, the mechanism of AHR transcriptional activation in retinal pigment cells has not been reported. Here, we demonstrate that the AHR signaling pathway is active in ARPE-19 cells, as in hepatocytes, but with different target gene specificity. We also found that chromatin remodeling by the BRG1-containing SWI/SNF complex is required for the AHR-mediated expression of target genes in ARPE-19 cells. We identified a novel enhancer region (−12 kb) of the CYP1A1 gene in ARPE-19 cells, to which both AHR and BRG1 are recruited in a ligand-dependent manner. BRG1 is associated with the AHR in ARPE-19 cells, and the C-terminal activation domain of the AHR directly interacts with BRG1. Furthermore, depletion of BRG1 caused a reduction in chromatin accessibility at the CYP1A1 enhancer. These results suggest that ARPE-19 cells possess an AHR-mediated transcription pathway with different target gene specificity, and that BRG1 is required for AHR-mediated transcription in ARPE-19 cells. [ABSTRACT FROM AUTHOR]

Highlights: [•] SO2 preconditioning reduced I/R-induced myocardial injury. [•] SO2 preconditioning upregulated myocardial H2S/CSE pathway. [•] SO2 preconditioning downregulated NO/iNOS pathway. [•] SO2 preconditioning significantly increased myocardial antioxidative capacity. [ABSTRACT FROM AUTHOR]

• The bilateral glymphatic profile of unilateral TLE patients with histologically proved hippocampal sclerosis (HS) was evaluated by the DTI-ALPS method. • In comparison to HCs, the TLE patients with HS showed significantly reduced DTI-ALPS indices bilaterally, and the reduction was more pronounced ipsilaterally. • In TLE patients with HS, the ipsilateral DTI-ALPS index was found slightly lower than the contralateral but without evident statistical significance. • In TLE patients with HS, the ipsilateral DTI-ALPS index was significantly inversely correlated with disease duration. In this study, the diffusion tensor imaging along perivascular space analysis (DTI-ALPS) technique was utilized to evaluate the functional changes in the glymphatic system of the bilateral hemispheres in patients with unilateral temporal lobe epilepsy (TLE) accompanied by hippocampal sclerosis (HS). The aim was to gain insights into the alterations in the glymphatic system function in TLE patients. A total of 61 unilateral TLE patients with HS and 53 healthy controls (HCs) from the Department of Neurosurgery at Xiangya Hospital were included in the study. All subjects underwent DTI using the same 3 T MR Scanner, and the DTI-ALPS index was calculated. Differences in the DTI-ALPS index between TLE patients and HCs were evaluated, along with the correlation between the DTI-ALPS index of TLE and clinical features of epilepsy. These features included age, age of onset, seizure duration, and neuropsychological scores. Compared to the bilateral means of the HCs, both the ipsilateral and contralateral DTI-ALPS index of the TLE patients were significantly decreased (TLE ipsilateral 1.41 ± 0.172 vs. HC bilateral mean: 1.49 ± 0.116, p = 0.006; TLE contralateral: 1.42 ± 0.158 vs. HC bilateral mean: 1.49 ± 0.116, p = 0.015). The ipsilateral DTI-ALPS index in TLE patients showed a significant negative correlation with disease duration (r = -0.352, p = 0.005). The present study suggests the presence of bilateral dysfunctions in the glymphatic system and also highlight a laterality feature in these dysfunctions. Additionally, the study found a significant negative correlation between the ipsilateral DTI-ALPS index and disease duration, underscoring the significance of early effective interventions and indicating potential for the development of innovative treatments targeting the glymphatic system. [ABSTRACT FROM AUTHOR]

Abstract: Duchenne muscular dystrophy (DMD) is a fatal genetic disease caused by the absence of the sarcolemmal protein dystrophin. Dilated cardiomyopathy leading to heart failure is a significant source of morbidity and mortality in DMD. We recently demonstrated amelioration of DMD heart disease in 16 to 20-m-old dystrophin-null mdx mice using adeno-associated virus (AAV) mediated micro-dystrophin gene therapy. DMD patients show severe heart disease near the end of their life expectancy. Similarly, mdx mice exhibit profoundly worsening heart disease when they reach beyond 21months of age. To more rigorously test micro-dystrophin therapy, we treated mdx mice that were between 21.2 and 22.7-m-old (average, 22.1±0.2months; N=8). The ∆R4-23/∆C micro-dystrophin gene was packaged in the cardiotropic AAV-9 virus. 5×1012 viral genome particles/mouse were delivered to mdx mice via the tail vein. AAV transduction, myocardial fibrosis and heart function were examined 1.7±0.2months after gene therapy. Efficient micro-dystrophin expression was observed in the myocardium of treated mice. Despite the robust dystrophin expression, myocardial fibrosis was not mitigated. Most hemodynamic parameters were not improved either. However, ECG abnormalities were partially corrected. Importantly, treated mice became more resistant to dobutamine-induced cardiac death. In summary, we have revealed for the first time the potential benefits and limitations of AAV micro-dystrophin therapy in end-stage Duchenne dilated cardiomyopathy. Our findings have important implications for the use of AAV gene therapy in dilated cardiomyopathy and heart failure. [Copyright &y& Elsevier]

Abstract: Duchenne muscular dystrophy (DMD) is a fatal genetic disease caused by the absence of the sarcolemmal protein dystrophin. Dilated cardiomyopathy leading to heart failure is a significant source of morbidity and mortality in DMD. We recently demonstrated amelioration of DMD heart disease in 16 to 20-m-old dystrophin-null mdx mice using adeno-associated virus (AAV) mediated micro-dystrophin gene therapy. DMD patients show severe heart disease near the end of their life expectancy. Similarly, mdx mice exhibit profoundly worsening heart disease when they reach beyond 21months of age. To more rigorously test micro-dystrophin therapy, we treated mdx mice that were between 21.2 and 22.7-m-old (average, 22.1±0.2months; N=8). The ∆R4-23/∆C micro-dystrophin gene was packaged in the cardiotropic AAV-9 virus. 5×1012 viral genome particles/mouse were delivered to mdx mice via the tail vein. AAV transduction, myocardial fibrosis and heart function were examined 1.7±0.2months after gene therapy. Efficient micro-dystrophin expression was observed in the myocardium of treated mice. Despite the robust dystrophin expression, myocardial fibrosis was not mitigated. Most hemodynamic parameters were not improved either. However, ECG abnormalities were partially corrected. Importantly, treated mice became more resistant to dobutamine-induced cardiac death. In summary, we have revealed for the first time the potential benefits and limitations of AAV micro-dystrophin therapy in end-stage Duchenne dilated cardiomyopathy. Our findings have important implications for the use of AAV gene therapy in dilated cardiomyopathy and heart failure. [Copyright &y& Elsevier]

Abstract: (-)-Epigallocatechin-3-gallate (EGCG) induces apoptosis in cancer cells without adversely affecting normal cells. Understanding the cancer-specific cytotoxic activity of EGCG is very important in defining the mechanism of tumorigenesis and identifying superb chemotherapeutic agents against cancer. We comparatively assayed human telomerase reverse transcriptase (hTERT)-mediated apoptosis by EGCG-induced reactive oxygen species (ROS) in normal cells and cancer cells. EGCG showed differential levels of ROS induction between the cell types; ROS, especially hydrogen peroxide, was highly induced in cancer cells, while it was not in normal cells. In addition, the higher level of ROS down-regulated hTERT via binding of CCCTC binding factor (CTCF) to the core promoter region of hTERT, which repressed hTERT expression. CTCF binding was epigenetically controlled by the demethylation of the previously hypermethylated site for CTCF, which was induced by down-regulation of DNA methyltransferase 1 (DNMT1). In contrast, hTERT down-regulation was not observed in normal cells. These results suggest that preferential death of cancer cells by EGCG could be caused by the cancer-specific induction of ROS and epigenetic modulation of expression of apoptosis-related genes, such as hTERT. [Copyright &y& Elsevier]

Abstract: Pulmonary arterial hypertension (PAH) is a serious disorder with poor prognosis. Urotensin II (UII) has been confirmed to be powerful vasoconstrictor than endothelin-1, which may play an important role in PAH development. The aim of this study is to observe the effects of urantide, a UII receptor antagonist， on monocrotaline (MCT) induced PAH in rats. 60 male Wistar rats were divided into six groups. For early treatment experiment, rats were divided into normal control group, MCT4w model group (MCT + saline × 3wks from the 8th day of MCT injection) and urantide early treatment group (MCT + urantide 10 μg/kg/d×3wks, 1 week after MCT injection once). For late treatment experiment, rats were divided as controls, MCT6w model group (MCT + saline×2wks, 4 weeks after MCT injection once) and urantide late treatment group (MCT + urantide 10 μg/kg/d×2wks, 4 weeks after MCT injection once). At the end of experiments, mean pulmonary arterial pressures (mPAP) and mean blood pressure (MBP) of rats in each group were measured by catheterization. Right ventricular weight ratio was also weighed. Relaxation effects of urantide on intralobar pulmonary arterial rings of normal control and MCT4w model rats were investigated. Pulmonary artery remodeling was detected by hematoxylin and eosin (HE) staining and immunohistochemistry analysis. Serum nitric oxide (NO) levels in all six groups were assayed by ELISA kits. Urantide markedly reduced the mPAP levels of MCT induced PAH in both early and late treatment groups. It didn’t change the MBP. Urantide dose-dependently relaxed the pulmonary arterial rings of normal control and MCT4w model rats. Moreover, NG-Nitro-l-arginine Methyl Ester (l-NAME) blocked the dilation response induced by urantide. In addition, urantide inhibited the pulmonary vascular remodeling remarkably. Serum NO level elevated in both early and late treatment rats with urantide infusion. These results suggest that urantide effectively alleviated MCT induced rats PAH may through relaxing pulmonary arteries and inhibiting pulmonary vascular remodeling. NO pathway might be one of the mechanisms in urantide induced pulmonary artery dilation. Thus, it is expected that urantide may be a novel therapy for PAH. [Copyright &y& Elsevier]

Abstract: LIGHT is a member of tumor necrosis factor (TNF) superfamily, and its function is mediated through lymphotoxin-β receptor (LTβR), which is known to play important roles in inflammatory and immune responses through activation of NF-κB signaling pathways. However, molecular mechanism of LTβR ligation-induced NF-κB signaling remains incompletely understood. In this report we demonstrate that a novel zinc-finger protein 91 (ZFP91) is a critical regulator in LIGHT-induced activation of non-canonical NF-κB pathway. ZFP91 appears to be required for NF-κB2 (p100) processing to p52, nuclear translocation of p52 and RelB, and DNA-binding activity of NF-κB in LIGHT-induced activation of non-canonical NF-κB pathway. Furthermore, ZFP91 knock-down by RNA interference blocks the LIGHT-induced accumulation of NIK and p100 processing, as well as the expression of non-canonical NF-κB target genes. These data clearly indicate that ZFP91 is a key regulator in LIGHT-induced activation of non-canonical NF-κB pathway in LTβR signaling. [ABSTRACT FROM AUTHOR]

Abstract: Hyperglycemia is the major cause of diabetic angiopathy. The aim of our study was to evaluate the impact of KB-R7943, an inhibitor of Na+/Ca2+ exchanger (NCX) on cell growth and function of human “diabetic” endothelial cells (EC). Intercellular adhesion molecule-1 (ICAM-1) expression and NCX activity were determined after EC were exposed to high glucose in the absence and presence of KB-R7943. Coincubation of EC with high glucose for 24h resulted in a significant increase of monocyte-endothelial cell adhesion and the expression of ICAM-1. These effects were abolished by KB-R7943 and KB-R7943 significantly decreased the activation of NCX induced by high glucose. These findings suggested that KB-R7943 may play a role in inhibiting expression of adhesion molecules by inhibiting the reverse activation of NCX. [Copyright &y& Elsevier]

Abstract: In order to find the biorational pesticides, we have synthesized two series of daphneolone analogues including 5-methylfuryl chalcones and 5-nitrofuryl chalcones by optimizing daphneolone analogues from Stellera chamaejasme L. with the good insecticidal properties of Qinghai-Tibet Plateau. All the synthesized compounds have been evaluated for anti-plant pathogenic fungi activities. The bioactivity assay showed that some of these daphneolone analogues were potentially active against plant pathogenic fungi, Rhizoctonia solani, Gibberella zeae, Bipolaris maydis, Sclerotia sclerotium and Botrytis cirerea, while the most potent 1-(2,4-dichlorophenyl)-3-(5-methylfuran-2-yl)propenone (compound 7) in this study showed good inhibitory activity against R. solani at 200mgL−1 with ca. 100% inhibition. [Copyright &y& Elsevier]

Abstract: As one of major epigenetic changes responsible for tumor suppressor gene inactivation in the development of cancer, promoter hypermethylation was proposed as a marker to define novel tumor suppressor genes. In the current study we identified ZIC1 (Zic family member 1, odd-paired Drosophila homolog) as a novel tumor suppressor gene silenced through promoter hypermethylation in gastric cancer, the second leading cause of cancer death worldwide. In all of gastric cancer cells lines examined, ZIC1 expression was downregulated and such downregulation was accompanied with the hypermethylation of ZIC1 promoter. Demethylation treatment with 5-aza-2′-deoxycytidine (Aza) reversed ZIC1 downregulation, highlighting the importance of promoter methylation to ZIC1 downregulation in gastric cancer cells. Notably, ZIC1 expression was significantly downregulated in primary gastric carcinoma tissues in comparison with non-tumor adjacent gastric tissues (p <0.01). Accordingly, promoter methylation of ZIC1 was frequently detected in primary gastric carcinoma tissues (94.6%, 35/37) but not normal gastric tissues, indicating that promoter hypermethylation mediated ZIC1 downregulation may play an important role in gastric carcinogenesis. Indeed, ectopic expression of ZIC1 led to the growth inhibition of gastric cancer cells through the induction of S-phase cell cycle arrest (p <0.01). Our results revealed ZIC1 as a novel candidate tumor suppressor gene downregulated through promoter hypermethylation in gastric cancer. [Copyright &y& Elsevier]

Abstract: Induction of cyclooxygenase-2 (COX-2) associates with cigarette smoke exposure in many malignancies. Nicotine and its derivative, 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), are the two important components in cigarette smoke that contributes to cancer development. However, the molecular mechanism(s) by which nicotine or NNK promotes gastric carcinogenesis remains largely unknown. We found that nicotine and NNK significantly enhanced cell proliferation in AGS cells that expressed both alpha7 nicotinic acetylcholine receptor (α7 nAChR) and β-adrenergic receptors. Treatment of cells with α-bungarotoxin (α-BTX, α7nAChR antagonist) or propranolol (β-adrenergic receptor antagonist) blocked NNK-induced COX-2/PGE2 and cell proliferation, while nicotine-mediated cell growth and COX-2/PGE2 induction can only be suppressed by propranolol, but not α-BTX. Moreover, in contrast to the dependence of growth promoting effect of nicotine on Erk activation, inhibitor of p38 mitogen-activated protein kinase (MAPK) repressed NNK-induced COX-2 upregulation and resulted in suppression of cell growth. In addition, nicotine and NNK mediated COX-2 induction via different receptors to modulate several G1/S transition regulatory proteins and promote gastric cancer cell growth. Selective COX-2 inhibitor (SC-236) caused G1 arrest and abrogated nicotine/NNK-induced cell proliferation. Aberrant expression of cyclin D1 and other G1 regulatory proteins are reversed by blockade of COX-2. These results pointed to the importance of adrenergic and nicotinic receptors in gastric tumor growth through MAPK/COX-2 activation, which may perhaps provide a chemoprevention strategy for cigarette smoke-related gastric carcinogenesis. [Copyright &y& Elsevier]

A eukaryotic-type signaling system in group A Streptococcus (GAS) was identified and characterized. This system comprises primarily the products of two co-transcribed genes, a eukaryotic-type Ser/Thr kinase (SP-STK) and phosphatase (SP-STP) and their endogenous substrate histone-like protein (SP-HLP). Enzyme activities of SP-STK and SP-STP primarily depended on Mn2+. The site on the substrate for reversible phosphorylation by these enzymes was found to be only the threonine residue. Using specific antibodies generated against these proteins, SP-STK was found to be membrane-associated with its N-terminal kinase domain facing the cytoplasm and its C-terminal repeat domain outside the membrane and cell-wall associated. Further, SP-STP, primarily a cytoplasmic protein, was found to be a major secretory protein of GAS and essential for bacterial survival. Three isogenic mutants, lacking either the entire SP-STK, or one of its two domains, were found displaying distinct pleiotropic effects on growth, colony morphology, cell division/septation, surface protein/virulence factor expression, bacterial ability to adhere to and invade human pharyngeal cells, and resist phagocytosis by human neutrophils. In addition to these properties, the ability of these three proteins to modulate the expression of the major virulence factors, the M protein and the capsule, indicates that these proteins are structurally and functionally distinct from the kinases and phosphatases described in other microorganisms and play a key role in GAS pathogenesis. [Copyright &y& Elsevier]

Streptococcal surface dehydrogenase (SDH) is a multifunctional, anchorless protein present on the surface of group A Streptococcus (GAS). It plays a regulatory role in GAS-mediated intracellular signaling events in human pharyngeal cells. Using ligand-binding assays, we have identified an ∼55kDa protein as an SDH-specific receptor protein on the surface of Detroit human pharyngeal cells. LC–MS/MS analyses identified this SDH-binding pharyngeal cell-surface-exposed membrane-bound protein as uPAR (urokinase plasminogen activator receptor)/CD87. Ligand-binding assays also revealed that only the N-terminal domain (D1) of uPAR bound to SDH. uPAR-D1 more specifically bound to the C-terminal alpha-helix and two immediate flanking regions of the S-loop of the SDH molecule. Site-directed mutagenesis in GAS resulting in SDH with altered C-terminal ends, and the removal of uPAR from pharyngeal cells by phosphatidylinositol-phopsholipase C treatment decreased GAS ability to adhere to pharyngeal cells. When compared to uninfected Detroit pharyngeal cells, GAS-infected pharyngeal cells showed a transient but a significant increase in the expression of uPAR-specific mRNA, and a prolonged recycling process of uPAR on the cell surface. Together, these results indicate that the specific streptococcal surface protein–pharyngeal cell receptor interaction mediated by SDH and uPAR is modulated during GAS infection of human pharyngeal cells. This interaction significantly contributes to bacterial adherence and thus may play a significant role in GAS pathogenesis by regulating intracellular signaling events in pharyngeal cells. [Copyright &y& Elsevier]

Abstract: The H3N2 vaccine strain (A/Panama/2007/99) for the 2003–2004 influenza season did not antigenically match the circulating A/Fujian/411/02-like H3N2 viruses and had reduced effectiveness against influenza outbreaks. A/Wyoming/03/2003, an A/Fujian-like virus, was recommended as the vaccine strain for the 2004–2005 season. A/Wyoming differed from A/Panama by 16 amino acids in the HA1 molecule. Reverse genetics was used to determine the minimal amino acid changes that were responsible for the antigenic drift from A/Panama to A/Wyoming. After substitutions of 2 of the 16 amino acids in the HA (H155T, Q156H), the A/Panama HA variant was antigenically equivalent to A/Wyoming as determined by hemagglutination inhibition and microneutralization assays using ferret postinfection antisera. Conversely, A/Wyoming containing the His-155 and Gln-156 residues from A/Panama was antigenically equivalent to A/Panama. These results indicated that only these two HA residues specified the antigenic drift from A/Panama to A/Wyoming; other amino acid differences between these two H3N2 viruses had minimal impact on virus antigenicity but impacted virus replication efficiency in eggs. [Copyright &y& Elsevier]

Segmental duplications account for up to 6% of the human genome, and the resulting low-copy repeats (LCRs) are known to be associated with more than 20 genomic disorders. Many such duplication events coincided with the burgeoning of the Alu repeat family during the last 50 million years of primate evolution, and it has been suggested that the two phenomena might be causally related. In tracing the evolution of the BRCA1 17q21 region through the primate clade, we discovered the occurrence over the last 40 million years of a complex set of about eight large gene-conversion-mediated rearrangements in the ∼4 Mb surrounding the BRCA1 gene. These have resulted in the presence of large and probably recombinogenic LCRs across the region, the creation of the NBR2 gene, the duplication of the BRCA1/NBR1 promoter, the bisection of the highly conserved ARF2 gene, and multiple copies of the KIAA0563 gene. The junctions lie within AluS repeats, members of an Alu subfamily which experienced massive expansion during the time that the rearrangements occurred. We present a detailed history of this region over a critical 40 million-year period of genomic upheaval, including circumstantial evidence for a causal link between Alu family expansion and the rearrangement-mediated destruction and creation of transcription units. [Copyright &y& Elsevier]

In cell and tissue samples, water is normally three orders of magnitude more abundant than other metabolites. Thus, water suppression is required in the acquisition of NMR spectra to overcome the dynamic range problem and to recover metabolites that overlap with the broad baseline of the strong water resonance. However, the heterogeneous cellular environment often complicates water suppression and the strong coupling of water to membrane lipids interferes with the NMR detection of membrane associated lipid components. The widely used water suppression techniques including presaturation and double pulsed field gradient selective echo result in more than a 70% reduction in membrane associated lipid components in proton spectra of cells and tissues compared to proton spectra acquired in the absence of water suppression. A water suppression technique based on the combination of selective excitation pulses and pulsed field gradients is proposed to use in the acquisition of high resolution MAS NMR spectra of tissue specimens and cell samples. This pulse sequence methodology enables efficient water suppression for intact cells and tissue samples and eliminates signal loss from cellular metabolites. [Copyright &y& Elsevier]

The purpose of this study is to determine whether angiostatin is involved in maintaining corneal avascularity after wounding.We generated polyclonal rabbit anti-mouse angiostatin antibodies directed against each of the five kringle domains, (K1-5) and anti-mouse plasmin B chain antibodies. Mouse corneas were immunostained with anti-K1 angiostatin antibody after excimer laser keratectomy. Corneal epithelial cell lysate was harvested and angiostatin was isolated using lysine sepharose. Purified plasminogen was incubated with lysate of mouse corneal epithelial cells from wild type mice in the presence or absence of MMP inhibitors. Angiostatin activity was determined using calf pulmonary artery endothelial (CPAE) cell proliferation assay with and without angiostatin immunoprecipitation; and corneal neovascularization was assayed by intrastromal injection of anti-plasminogen, anti-K1-3 or anti-B chain antibodies after corneal wounding.Using the anti-mouse angiostatin antibodies that we generated, we confirmed that angiostatin-like molecules were expressed in the corneal epithelium and in cultured corneal epithelial cells. Western blotting after incubation of scraped corneal epithelial cell lysate with purified plasminogen showed reduction of the plasminogen bands at 6, 12, and 24 hr, respectively. Complete cleavage of plasminogen occurred by 48 hr. Functional assays in which corneal epithelial cell extracts were incubated with CPAE cells resulted in inhibition of vascular endothelial cell proliferation. Depletion experiments using anti-angiostatin (K1) antibodies resulted in a 25±1·2% increase in vascular endothelial cell proliferation as compared to 12±1·8% using the protein A control (p<0·05). Corneal neovascularization was observed after excimer laser keratectomy when anti-angiostatin antibodies were injected into the cornea (65±13%) which was significantly higher than when plasmin B chain antibodies were injected (10±2·6%; p<0·05).Plasminogen and angiostatin are produced in the cornea. They may play a role in preventing vascularization and may contribute to the maintenance of corneal avascularity after excimer laser keratectomy. [Copyright &y& Elsevier]

FluMist influenza A vaccine strains contain the PB1, PB2, PA, NP, M, and NS gene segments of ca A/AA/6/60, the master donor virus–A strain. These gene segments impart the characteristic cold-adapted (ca), attenuated (att), and temperature-sensitive (ts) phenotypes to the vaccine strains. A plasmid-based reverse genetics system was used to create a series of recombinant hybrids between the isogenic non-ts wt A/Ann Arbor/6/60 and MDV-A strains to characterize the genetic basis of the ts phenotype, a critical, genetically stable, biological trait that contributes to the attenuation and safety of FluMist vaccines. PB1, PB2, and NP derived from MDV-A each expressed determinants of temperature sensitivity and the combination of all three gene segments was synergistic, resulting in expression of the characteristic MDV-A ts phenotype. Site-directed mutagenesis analysis mapped the MDV-A ts phenotype to the following four major loci: PB11195 (K391E), PB11766 (E581G), PB2821 (N265S), and NP146 (D34G). In addition, PB12005 (A661T) also contributed to the ts phenotype. The identification of multiple genetic loci that control the MDV-A ts phenotype provides a molecular basis for the observed genetic stability of FluMist vaccines. [Copyright &y& Elsevier]

In the present study, the concentrations of lead (Pb), cadmium (Cd), arsenic (As), mercury (Hg), and copper (Cu) in 2245 batches of Chinese herbal medicines (CHMs) were measured using inductively coupled plasma-mass spectroscopy (ICP-MS). We developed a risk assessment strategy that assessed the heavy metal-associated health risk of CHMs based on our large dataset. Using a combination of the mean and 95th percentile (P95) values of the chronic daily intake (CDI), hazard quotient (HQ), hazard index (HI), and lifetime cancer risk (CR), the health risks of the average exposure population and the high exposure population were estimated, respectively. To obtain a precise and realistic risk assessment, the exposure frequency and exposure duration were determined using questionnaire data from 20,917 randomly selected volunteers. Additionally, given the specific ingestion characteristics of CHMs, the safety factor and the transfer rates of heavy metals were highlighted as well. The concentrations of Pb, Cd, As, Hg, and Cu in 2245 batches of CHMs were 1.566, 0.299, 0.391, 0.074, and 8.386 mg/kg, respectively. The mean HI values indicated that consumption of most CHMs would not pose an unacceptable health risk to the average exposure population, except for argy wormwood leaf (1.326), morinda root (2.095), plantain herb (1.540), chrysanthemum flower (1.146), and Indian madder root (2.826). In addition, CR assessment for Pb and As revealed that, for the average exposure population, the risk of developing cancers was lower than the acceptable levels (1 × 10−4) in the clinic. However, the P95 of the HI and CR values indicated that more attention should be paid to the systemic effects of CHMs in terms of both non-carcinogenic and carcinogenic health risks for the high exposure population. Furthermore, in order to serve population health better, national and international guidelines have now been established. The risk assessment strategy developed in this study is the first of its kind, and contributed to the risk assessment, guidelines, and safety standards for heavy metals in CHMs. [ABSTRACT FROM AUTHOR]

[Display omitted] • Co-NCNT@HC was proposed to enhance lithium sulfur batteries. • Interconnected hollow carbon cages with cobalt and nitrogen decoration. • The LSBs present high stability and long lifespan over 1000 cycles. Lithium-sulfur (Li-S) battery has been considered as a potential next-era energy storage device. However, its practical application is limited by the volume change of sulfur and the shuttle effect of lithium polysulfides. To effectively overcome these issues, a hollow carbon decorated with cobalt nanoparticles and interconnected by nitrogen doped carbon nanotubes (Co-NCNT@HC) is developed for high-performance Li-S battery. The uniformly distributed nitrogen and cobalt nanoparticles in Co-NCNT@HC are able to enhance the chemical adsorption capability and fasten the transformation speed of the intermediates, thus effectively inhibit the loss of lithium polysulfides. Moreover, the hollow carbon spheres interconnected by carbon nanotubes are structurally stable and electrically conductive. Due to the unique structure, the Li-S battery enhanced by Co-NCNT@HC shows a high initial capacity of 1550 mAh/g at 0.1 A g−1. Even at a high current density of 2.0 A g−1, after 1000 cycles, it still maintains a capacity of 750 mAh/g with a capacity retention of 76.4% (the capacity decay rate is only 0.037% per cycle). This study provides a promising strategy for the development of high-performance Li-S batteries. [ABSTRACT FROM AUTHOR]

Recently, we reported that Ilexgenin A exhibits anti-cancer activities and induces cell arrest. Here, we investigated the effect of Ilexgenin A on the inflammation, angiogenesis and tumor growth of hepatocellular carcinoma (HCC). Our current study revealed that Ilexgenin A significantly inhibited the inflammatory cytokines TNF-α and IL-6 levels and downregulated pro-angiogenic factor VEGF production and transcription in HepG2 cells. The underlying mechanism for Ilexgenin A effects appears to be through inhibiting STAT3 and PI3K pathways. Furthermore, we found that not only Ilexgenin A inhibited STAT3 and PI3K pathways in HepG2 cells but also blocked these signaling pathways in HUVECs. Most importantly, by employing two HCC xenografts models - HepG2 and H22, we showed that Ilexgenin A reduced tumor growth and exhibited synergy effect with Sorafenib. ELISA assay, histological analysis and immunohistochemistry examination revealed that the expression of VEGF and MVD was significantly decreased after the treatment with Ilexgenin A and the combination. Moreover, Ilexgenin A could enhance caspase-3/7 activity in vitro and transmission electron microscope indicated that the combination induced evident apoptosis of tumor cells and caused the structural changes of mitochondria in vivo. Although no apparent adverse effects occurred during the treatment period, Sorafenib monotherapy elicited hepatotoxicity for specific expression in the increased level of AST and the ratio of AST/ALT. However, the combination could remedy this adverse effect. In conclusion, the results described in the present study identifies Ilexgenin A as a promising therapeutic candidate that modulates inflammation, angiogenesis, and HCC growth. [ABSTRACT FROM AUTHOR]

• Pleural CXCL9 and CXCL11 are potential biomarkers for diagnosing TPE. • The diagnostic accuracy of CXCL9 and CXCL11 are affected by age. • CXCL9 and CXCL11 promote peripheral T helpers migrating to the pleural cavity, thus representing therapeutic targets for TPE treatment. Pleural biomarkers represent potential diagnostic tools for tuberculous pleural effusion (TPE) due to their advantages of low cost, short turnaround time, and less invasiveness. This study evaluated the diagnostic accuracy of two CXCR3 ligands, C-X-C motif chemokine ligand 9 (CXCL9) and CXCL11, for TPE. In addition, we investigated the cellular origins and biological roles of CXCL9 and CXCL11 in the development of TPE. This double-blind study prospectively enrolled patients with undiagnosed pleural effusion from two centers (Hohhot and Changshu) in China. Pleural fluid on admission was obtained and levels of CXCL9 and CXCL11 were measured by an enzyme-linked immunosorbent assay (ELISA). The receiver operating characteristic (ROC) curve and the decision curve analysis (DCA) were used to evaluate their diagnostic accuracy and net benefit, respectively. THP-1 cell-derived macrophages were treated with Bacillus Calmette-Guérin (BCG), and quantitative real-time PCR (qRT-PCR) and ELISA were used to determine the mRNA and protein levels of CXCL9 and CXCL11. The chemoattractant activities of CXCL9 and CXCL11 for T helper (Th) cells were analyzed by a transwell assay. One hundred and fifty-three (20 TPEs and 133 non-TPEs) patients were enrolled in the Hohhot Center, and 58 (13 TPEs and 45 non-TPEs) were enrolled in the Changshu Center. In both centers, we observed increased CXCL9 and CXCL11 in TPE patients. The areas under the ROC curves (AUCs) of pleural CXCL9 and CXCL11 in the Hohhot Center were 0.70 (95 % CI: 0.55–0.85) and 0.68 (95 % CI: 0.52–0.84), respectively. In the Changshu Center, the AUCs of CXCL9 and CXCL11 were 0.96 (95 % CI: 0.92–1.00) and 0.97 (95 % CI: 0.94–1.00), respectively. The AUCs of CXCL9 and CXCL11 decreased with the advancement of age. The decision curves of CXCL9 and CXCL11 showed net benefits in both centers. CXCL9 and CXCL11 were upregulated in BCG-treated macrophages. Pleural fluid from TPE and conditioned medium from BCG-treated macrophages were chemotactic for Th cells. Anti-CXCL9 or CXCL11 neutralizing antibodies could partly block the chemotactic activity. Pleural CXCL9 and CXCL11 are potential diagnostic markers for TPE, but their diagnostic accuracy is compromised in elderly patients. CXCL9 and CXCL11 can promote the migration of peripheral Th cells, thus representing a therapeutic target for the treatment of TPE. [ABSTRACT FROM AUTHOR]

Osteoarthritis (OA) is an incurable joint disease affecting 240 million elderly population, and major unmet medical needs exist for better therapeutic options for OA. During skeletal development, Nkx3.2 has been shown to promote chondrocyte differentiation and survival, but to suppress cartilage hypertrophy and blood vessel invasion. Here we show that Nkx3.2 plays a key role in osteoarthritis (OA) pathogenesis. Marked reduction of Nkx3.2 expression was observed in three different murine OA models. Consistent with these findings, analyses of surgery-induced and age-driven OA models revealed that cartilage-specific post-natal induction of Nkx3.2 can suppress OA progression in mice. These results suggest that Nkx3.2 may serve as a promising target for OA drug development. [Display omitted] • Nkx3.2 expression is found to be reduced in multiple osteoarthritis (OA) models. • Nkx3.2 can alleviate disease severity in both traumatic and degenerative OA models. • Nkx3.2 can protect articular cartilages from osteoarthritic destruction. • Nkx3.2 can attenuate osteophyte maturation facilitated by aging. • Nkx3.2 may serve as a promising target for OA drug development. [ABSTRACT FROM AUTHOR]

At present, stem cell transplantation has a significant therapeutic effect on spinal cord injury (SCI), however, it is still challenging for the seed cells selection. In this study, in order to explore cells with wide neural repair potentials, we selected the pluripotent stem cells multilineage-differentiating stress-enduring (Muse) cells, inducing the in vitro differentiation of human Muse cells into neural precursor cells (Muse-NPCs) by applying neural induction medium. Here, we found induced Muse-NPCs expressed neural stem cell markers Nestin and NCAM, capable of differentiating into three types of neural cells (neuron, astrocyte and oligodendrocyte), and have certain biological functions. When Muse-NPCs were transplanted into rats suffering from T10 SCI, motor function was improved. These results provide an insight for application of Muse-NPCs in cell therapy or tissue engineering for the repair of SCI in future. • Muse cells can be induced into neural precursor cells in vitro. • Muse-NPCs different into neurons, astrocytes and oligodendrocytes with their own functions in vitro. • Muse-NPCs promote the restoration of motor function in SCI rats. [ABSTRACT FROM AUTHOR]

Abstract: 3-Hydroxyalkanoic acids (3HA) are precious precursors for synthesis of value added chemicals. According to their carbon chain lengths, 3HA can be divided into two groups: short-chain-length (SCL) 3HA consisting of 3–5 carbon atoms and medium-chain-length (MCL) 3HA containing 6–14 carbon atoms. To produce MCL 3HA, a metabolic engineered pathway expressing tesB gene, a thioesterase encoding gene that has been reported to catalyze acyl-CoA to free fatty acids, was constructed in Pseudomonas entomophila L48. When tesB of Escherichia coli encoding thioesterase II was introduced into polyhydroxyalkanoate (PHA) synthase and β-oxidation pathway deleted mutant of P. entomophila LAC31 derived from wild type P. entomophila L48, 6.65g/l 3-hydroxytetradecanoic acid (3HTD) and 4.6g/l 3-hydroxydodecanoic acid (3HDD) were obtained, respectively, when tetradecanoic acid or dodecanoic acid as related carbon sources was added in shake flask cultures. Moreover, 1.8g/l of 3-hydroxydecanoic (3HD) acid was also produced by P. entomophila LAC31 harboring PTE1 gene cloned from Saccharomyces cerevisiae using corresponding fatty acid decanoic acid. Interestingly, shake flask studies indicated that PTE1 harboring strain showed advantages over tesB expressing one for 3HDD and 3HD production, while tesB favored 3HTD production by P. entomophila LAC31. For the first time our study revealed that fine chemicals 3HTD, 3HDD or 3HD could be efficiently produced by metabolic engineered β-oxidation in Pseudomonas spp grown on related fatty acids. [Copyright &y& Elsevier]

Abstract: We studied the attenuation, immunogenicity and efficacy of the cold-adapted A/Ann Arbor/6/60 (AA ca) (H2N2) virus in mice and ferrets to evaluate its use in the event of an H2 influenza pandemic. The AA ca virus was restricted in replication in the respiratory tract of mice and ferrets. In mice, 2 doses of vaccine elicited a >4-fold rise in hemagglutination–inhibition (HAI) titer and resulted in complete inhibition of viral replication following lethal homologous wild-type virus challenge. In ferrets, a single dose of the vaccine elicited a >4-fold rise in HAI titer and conferred complete protection against homologous wild-type virus challenge in the upper respiratory tract. In both mice and ferrets, the AA ca virus provided significant protection from challenge with heterologous H2 virus challenge in the respiratory tract. The AA ca vaccine is safe, immunogenic, and efficacious against homologous and heterologous challenge in mice and ferrets, supporting the evaluation of this vaccine in clinical trials. [Copyright &y& Elsevier]

Abstract: An economical mesoporous aluminosilicate was synthesized with microcline as starting material and the precursor 13X zeolite as seed for crystal structure on mesoporous walls. In this method, a mixture of microcline and Na2CO3 with a molar ratio of 1:1.05 was first calcined at 1093 K for 2.5 h. The calcined materials were mixed with 35 ml C16TMABr aqueous solution (containing 8.2 g C16TMABr) and the precursors of 13X zeolite, resulting in mesoporous aluminosilicate after crystallization of the solution at 378 K for 48 h and calcination of the powder at 823 K for 5 h. The as-synthesized sample has a uniform pore diameter distribution centered at 3.7 nm. The as-synthesized sample had BET surface area of 725 m2/g and BJH mean pore diameter of 3.7 nm. The FT-IR results revealed that the building units of 13X zeolite were inserted into the pore walls of the as-synthesized sample. The adsorption ratio of mercury(II) onto the as-synthesized adsorbent was about 95%. The adsorption process was found to be spontaneous and can be explained by particle diffusion and chemical ion-exchange mechanisms. The equilibrium concentration of mercury(II) using the as-synthesized sample as the adsorbent was under 1 μg/L, making the concentration of mercury meet the limit for drinking water in China as recommended by the World Health Organization. [Copyright &y& Elsevier]

Abstract: The present study examined the relationship between word concreteness and word frequency using event-related potential (ERP) measurements during a lexical decision task. Potential effects of concreteness in the processing of verbs were also examined. ERPs were recorded from 119 scalp electrodes in 23 right-handed participants. The results showed that concrete nouns were associated with a more negative ERP than abstract nouns at 200–300 and 300–500ms after stimulus onset, regardless of word frequency. Between 300 and 500ms, concrete nouns and abstract nouns produced differentiated scalp distributions, respectively. In terms of verbs, concreteness only produced small difference in ERP primarily in the central–parietal sites of the left hemisphere. [Copyright &y& Elsevier]

Abstract: Objective : Cisplatin (DDP) resistance is a major barrier to overcome before chemotherapy can become curative for most patients presenting with ovarian cancer. In this study, we investigated the effect of siRNAs on expression of p-gp, GST-pi mRNA and protein in cisplatin-resistant human ovarian cancer cells in order to restore sensitivity to DDP. Methods : Small interfering double-stranded RNAs (siRNA) were designed to target p-glycoprotein (p-gp) and glutathione S-transferases (GST) mRNA as a strategy to inhibit both resistant gene expression at the mRNA level. Using Real-Time PCR and western blotting assay the changes of the RNA and protein levels of both drug resistant genes were studied. Results : Transfection of MDR-1 and GST siRNAs into human multi-drug resistance (MDR) ovarian cancer cell lines, COC1/DDP and SKOV3/DDP, resulted in a time-dependent inhibition of both gene expressions with the decline of the IC50 values but had no effect on the expression of a-Tubulin. Inhibition of P-gp and GST expression by siRNA enhanced the intracellular accumulation of and restored sensitivity to DDP. Conclusions : These studies suggest that p-gp and GST siRNAs are effective inhibitors of MDR gene expression and reverse the resistance of ovarian carcinomas. Our studies may provide a new insight to develop siRNAs as a novel therapeutic tool for the treatment of ovarian carcinomas. [Copyright &y& Elsevier]

Alpha-enolase (SEN) is a strong plasminogen-binding protein on the surface of group A streptococci (GAS). By flow cytometry and immunofluorescence analyses and using human enolase-specific antibody, human pharyngeal cells (Detroit 562) also were found to express enolase on their surface. Detroit 562 cells preferentially bound to Lys-plasminogen and this binding was inhibited in the presence of a lysine analog, &epsiv;-aminocaproic acid and by carboxypeptidase-B treatment suggesting that the C-terminal lysine residue of the putative pharyngeal cell receptor(s) may play an important role in plasminogen-binding. The increased plasminogen-binding in the presence of free enolase indicated the presence of an enolase/SEN-specific receptor on the pharyngeal cell surface. GAS, when precoated with Lys-plasminogen, adhered to pharyngeal cells significantly more in numbers than when precoated with fibronectin or laminin. Similarly, GAS adhered also significantly more in numbers to pharyngeal cells which were precoated with Lys-plasminogen. GAS adhered similarly in high numbers when incubated with pharyngeal cells in the presence of soluble plasminogen. The de novo pharyngeal cell-bound protease activity, created as a result of activation of bound plasminogen by t-PA, indicated its potential role in pericellular fibrinolytic activity. Further GAS with tPA-activated plasminogen bound on their surface penetrated through Transwell-grown pharyngeal cells in significantly higher numbers. Together, the results presented in this study highlight a novel function of plasminogen in streptococcal adherence to pharyngeal cells and a newly discovered streptococcal ability to pericellularly invade pharyngeal cells as a result of tPA/endogenous plasminogen activator-mediated proteolytic activity. [Copyright &y& Elsevier]

Hepatocyte growth factor (HGF) is a versatile neurotrophic factor that mediates a variety of cellular activities. In this study, we investigated the effects of intramuscularly injected recombinant AAV vectors expressing HGF in two pathologic conditions: the sciatic nerve crush and the SOD1-G93A transgenic mouse models. AAV serotype 6 (rAAV6) was chosen based on its expression levels in, and capability of moving to, the spinal cord from the injected muscle area. In the nerve crush model, rAAV6-HGF was shown to reduce the degree of mechanical allodynia, increase the cross-sectional area of muscle fibers, promote regrowth of peripheral axons, and improve motor functions. In the SOD1-G93A TG mouse model, rAAV6-HGF increased the mass of the tibialis anterior and gastrocnemius, alleviated disease symptoms, and prolonged survival. Improvements in integrity and functions of muscle in these models seemed to have come from the ability of HGF produced from rAAV6-HGF to regulate the expression of various atrogenes through the control of the FOXO signaling pathway. Our findings suggested that intramuscular injection of rAAV6-HGF might be used to relieve various symptoms associated with muscle atrophy and/or nerve damages observed in a majority of neuromuscular diseases. • rAAV6 produced considerable amounts of the transgene product and moved to the spinal cord when injected intramuscularly. • rAAV6-HGF improved sensory and motor functions, and restored muscle integrity in the sciatic nerve crush mouse model. • rAAV6-HGF increased the muscle mass and alleviated symptoms of SOD1-G93A transgenic mice. [ABSTRACT FROM AUTHOR]

• GD neurons in DVC could be excited by microinjecting ghrelin into DVC and electrically stimulating LHA. • The excitatory effect was completely or partially blocked by [D-Lys-3]-GHRP-6 in the DVC. • The excitatory effect was weaker in cisplatin-treated rats than that in saline-treated rats. Cisplatin is an important antineoplastic drug and has side effects such as nausea, vomiting, and dyspepsia. The detailed mechanisms for its side effects are yet not well be illustrated. Our purpose was to investigate the discharges of gastric distention (GD) sensitive neurons regulated by ghrelin and electrical stimulation of the lateral hypothalamus area (LHA) via the dorsal vagal complex (DVC) in cisplatin-treated rats. Extracellular discharge recording was performed to observe the effects of ghrelin and electrical stimulation of the LHA on discharges of GD neurons in the DVC. GD neurons were recorded in DVC in saline-treated and cisplatin-treated rats and identified as GD-excitatory (GD-E) neurons, which are excited by gastric distension, and GD-inhibitory (GE-I) neurons, which are inhibited by gastric distension. Microinjection of ghrelin into the DVC increased the firing frequency of most GD neurons, while the ratios of excited GD-E and GD-I neurons in cisplatin-treated rats were significantly lower than those in saline-treated rats. The excitatory effect of ghrelin was eliminated completely by DVC pretreatment with ghrelin receptor antagonist [D-Lys-3]-GHRP-6. After electrical stimulation of the LHA, the firing frequency of these neurons significantly increased. This excitatory effect was weaker in cisplatin-treated rats than in saline-treated rats and could be partly blocked by DVC pretreatment with [D-Lys-3]-GHRP-6. GD neurons in the DVC could be excited by microinjecting ghrelin into the DVC and electrical stimulation of the LHA, respectively. The excitatory effect was attenuated by cisplatin injected intraperitoneally. [ABSTRACT FROM AUTHOR]

• The amplitude of out-put signal reflects the torque and frequency reflects the rotational speed. • It solves the problem of power supply and signal output of sensitive elements on the shaft. • This sensing method cleverly uses the non-contact principle of capacitance. Torque and rotational speed are important parameters to characterize the power of rotating parts for mechanical systems, so a rotational shaft's torque and rotational speed sensing method and real-time signal processing are key technologies for mechanized equipment, such as vehicles, ships, etc. Hence an advanced torque and speed sensing technology and a data processing technology are very important for improving the performance of mechanical equipment. However, the power transmission device's rotational shaft has a compact structure and narrow space, so it is difficult to install a signal transmission and power supply device of a sensor on the rotational shaft. To solve these difficult problems, this paper proposes a rotational shaft torque and rotational speed measurement method coupling resistance and capacitance. This method uses two resistance strain gauges and a capacitive grating, and the capacitive grating transmits signals in a non-contact coupling manner. The sensor on the rotational shaft needs no power supply and the signals are transmitted to a measurement circuit from a stationary sleeve, so this method essentially solves the difficult problems in signal transmission and power supply. Simulation and measurement experiments show that the resistance strain gauges obtain information and the capacitive grating sensor transfers information. The method coupling resistance and capacitance for realizing information sensing is feasible, which is suitable for measuring the torque and rotational speed of the rotational shaft of mechanical equipment in spite of the compact structure and narrow space. [ABSTRACT FROM AUTHOR]

Succinate dehydrogenase (SDH), an essential component of cellular respiratory chain and tricarboxylic acid (or Krebs) cycle, has been identified as one of the most significant targets for pharmaceutical and agrochemical. Herein, with the aim of discovery of new antifungal lead structures, a class of novel N -(4-fluoro-2-(phenylamino)phenyl)-pyrazole-4-carboxamides were designed, synthesized and evaluated for their biological activities. They were bioassayed against seven phytopathogenic fungi, Rhizoctonia solani , Phytophthora infestans , Fusarium oxysporum f. sp. vasinfectum , Botryosphaeria dothidea , Gibberella zeae , Alternaria alternate and Fusarium oxysporum f. sp. niveum. The results indicated that most of the compounds displayed good antifungal activities, especially against R. solani. Among them, compounds 7 and 12 exhibited higher antifungal activities against R. solani in vitro with EC 50 value of 0.034 mg/L and 0.021 mg/L, being superior to the commercially available fungicide bixafen (EC 50 = 0.043 mg/L). Pot tests against R. solani showed that in vivo EC 50 values of compounds 7 (2.694 mg/L) and 12 (2.331 mg/L) were higher than that of bixafen (3.724 mg/L). In addition, inhibitory activity of compound 12 against SDH indicated compound 12 (IC 50 = 1.836 mg/L) showed good inhibitory activity against SDH, being close to bixafen's inhibitory activity (IC 50 = 1.222 mg/L). And, molecular modeling of the SDH-compound 12 complex suggested that compound 12 could strongly bind to and interact with the binding site of the SDH. The results of the present work showed that N -(4-fluoro-2-(phenylamino)phenyl)-pyrazole-4-carboxamides were a new fungicides for discovery of SDH inhibitors and worth further study. Unlabelled Image • Twenty five novel N -(4-fluoro-2-(phenylamino)phenyl)-pyrazole-4-carboxamides were designed and synthesized. • Compound 12 exhibited the highest antifungal activities against R. solani in vitro and in vivo , being superior to bixafen. • Inhibitory activity against SDH indicated compound 12 showed good inhibitory activity against SDH, being close to bixafen. • Molecular modeling suggested that can compound 12 strongly bind to and interact with the binding site of the SDH. [ABSTRACT FROM AUTHOR]

To explore the toxicity mechanisms of neochamaejasmin A (NCA), extracted from Stellera chamaejasme L., we first evaluated its cytotoxicity on the Spodoptera frugiperda (Sf9) cell line. The results confirmed that NCA inhibited Sf9 cell survival in both a dose- and time-dependent manner. Then, intracellular biochemical assays showed that NCA induced apoptosis in Sf9 cells. Evidence of apoptosis was confirmed by morphological changes and the activation of caspases-3/9. We also observed that NCA induced apoptosis via mitochondrial-dependent intrinsic apoptotic pathway by upregulating cytochrome c and proapoptotic protein (Bax) and downregulating the mitochondrial membrane potential (MMP) and antiapoptotic protein (Bcl-2). Moreover, we found a dose-dependent increase in reactive oxygen species (ROS), accumulation of lipid peroxidation product and an inactivation of the antioxidant enzymes in treated cells. Additionally, the cleavage of PARP and G2/M arrest were also detected in Sf9 cells exposed to NCA. These findings provide critical information that NCA effectively induced apoptosis in Sf9 cells through mitochondrial pathways. Unlabelled Image • NCA could inhibit insect cell line Sf9. • NCA triggered apoptosis in Sf9 cells by mitochondrial pathway depending dose manner. • NCA could elevate ROS levels of Sf9 in a dose dependent manner. • NCA could arrest cell cycle of Sf9 in a dose dependent manner. [ABSTRACT FROM AUTHOR]

The high quality anodic aluminum oxide (AAO) defective photonic crystals (DPCs) have been successfully prepared by using a modified periodic pulse anodization technique including an effective voltage compensating strategy. The test results confirmed that the AAO DPCs were with a perfect regular layered-structure and had a narrow defective photonic band gap (DPBG) with a high quality defective mode. When the rhodamine B (rhB) was absorbed onto the pore walls of the AAO DPCs, it was found that the DPBG blue edge and localized defective mode inside could significantly enhance the photoluminescence (PL) intensity of rhodamine B (rhB), while they were carefully regulated to match with the emission peak position of rhB respectively. Even more intriguing was that the localized defective peak in DPBG had more notable effect on rhB’s photoluminescence, 3.1 times higher than that of the control samples under the same conditions. The corresponding mechanism for photoluminescence enhancement was also discussed in detail. [ABSTRACT FROM AUTHOR]

Insulin resistance, a pathophysiology of type 2 diabetes, is associated with obesity. Lipotoxicity in obesity leads to the dysfunction and death of pancreatic β-cells and inadequate insulin production, thereby aggravating type 2 diabetes. The present study was conducted to determine the effect of Aloe vera polysaccharides (APs) as an anti-hyperglycemic agent and their mechanisms of action. Gel polysaccharides from Aloe extracts were separated using ultrafiltration devices with molecular weight-cutoff membranes, and the protective effect of APs on pancreatic β-cells in response to free fatty acids (FFAs) was determined. Hamster pancreatic β-cell line HIT-T15 was treated with palmitate and APs to analyze cellular responses. We observed a large number of apoptotic β-cell death after treatment with high levels of palmitate, but this was efficiently prevented by the addition of APs in a dose-dependent manner. It was found that the anti-apoptotic properties of APs were largely due to the relief of endoplasmic reticulum (ER) stress signaling. APs were effective in interfering with the FFA-induced activation of the PERK and IRE1 pathways as well as ROS generation, thereby protecting pancreatic β-cells from lipotoxicity. Although variation in the chain length of APs can influence the activity of FFA-mediated ER stress signaling in different ways, polysaccharide mixtures with molecular weights higher than 50 kDa showed greater antiapoptotic and antioxidant activity in β-cells. After oral administration of APs, markedly lowering fasting blood glucose levels were observed in db/db mice, providing evidence of the potential of APs as an alternative insulin sensitizer. Therefore, it was concluded that APs have a protective effect against type 2 diabetes by modulating obesity-induced ER stress in pancreatic β-cells. [ABSTRACT FROM AUTHOR]

The present work aims to explore the time-response (from 24 h to 96 h) characteristic and identify early potential sensitive biomarkers of copper (Cu) (as copper chloride dihydrate), cadmium (Cd) (as cadmium acetate), lead (Pb) (as lead nitrate) and chromium (Cr) (as potassium dichromate) exposure in adult zebrafish, focusing on reactive oxygen species (ROS), SOD activity, lipid peroxidation and gene expression related to oxidative stress and inflammatory response. Furthermore, the survival rate decreased apparently by a concentration-dependent manner after Cu, Cr, Cd and Pb exposure, and we selected non-lethal concentrations 0.05 mg/L for Cu, 15 mg/L for Cr, 3 mg/L for Cd and 93.75μg/L for Pb to test the effect on the following biological indicators. Under non-lethal concentration, the four heavy metals have no apparent histological change in adult zebrafish gills. Similar trends in ROS production, MDA level and SOD activity were up-regulated by the four heavy metals, while MDA level responded more sensitive to Pb by time-dependent manner than the other three heavy metals. In addition, mRNA levels related to antioxidant system (SOD1, SOD2 and Nrf2) were up-regulated by non-lethal concentration Cu, Cr, Cd and Pb exposure. MDA level and SOD1 gene have a more delayed response to heavy metals. Genes related to immunotoxicity were increased significantly after heavy metals exposure at non-lethal concentrations. TNF-α and IL-1β gene have similar sensibility to the four heavy metals, while IL-8 gene was more responsive to Cr, Cd and Pb exposure at 48 h groups and IFN-γ gene showed more sensitivity to Cu at 48 h groups than the other heavy metals. In conclusion, the present works have suggested that the IFN-γ gene may applied as early sensitive biomarker to identify Cu-induced toxicity, while MDA content and IL-8 gene may use as early sensitive biomarkers for evaluating the risk of Pb exposure. Moreover, IL-8 and IFN-γ gene were more responsive to heavy metals, which may become early sensitive and potential biomarkers for evaluating inflammatory response induced by heavy metals. This work reinforces the concept of the usefulness of gene expression assays in the evaluation of chemicals effects and helps to establish a background data as well as contributes to evaluate early environmental risk for chemicals, even predicting toxicity. [ABSTRACT FROM AUTHOR]

Complement-mediated inflammation and tissue damage is an important drive to pathology in autoimmune diseases, targeting inhibit the complement activation is promising treatment strategy for these diseases. We performed anticomplement activity-guided fractionation of the water extract of Eclipta prostrata by ion-exchange and size-exclusion chromatography, yielding two bioactive polysaccharides EAP20-1 and EAP20-2. The molecular weights of EAP20-1 and EAP20-2 were respectively calculated to be 5.2 kDa and 6.3 kDa by HPGPC, both polysaccharides was composed by d -Gal, l -Glc, and Ara at different ratios of 7.3:2.7:1 and 7.6:3.1:1. In addition, the main linkage types of EAP20-1 and EAP20-2 were β-1,4-Gal, β-1,6-Gal and α-1,4,6-Glc according to methylation analyses. EAP20-1 and EAP20-2 exhibited significant inhibitory effect on the complement activation through both classical and alternative pathways and with no influence on the coagulation system. Preliminary mechanism study indicated that both EAP20-1 and EAP20-2 inhibited the activation of the complement system by interacting with C1q, C1r, C1s, C2, C4, C5, C7, and C9 components. [ABSTRACT FROM AUTHOR]

The effects of hot water blanching (HWB), microwave blanching (MWB), infrared blanching (IRB) and high-humidity hot air impingement blanching (HHAIB) on weight loss, enzyme inactivation (peroxidase, POD and polyphenol oxidase, PPO), characteristic bioactive component contents (red pigments and ascorbic acid), antioxidant capacity (total antioxidant activity and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging capacity), ultrastructure and drying kinetics of red bell peppers under different blanching conditions were investiaged. Results showed that the MWB and IRB treatment yielded lower weight loss than HWB and HHAIB. The inactivation of both PPO and POD enzymes was strongly dependent on blanching temperature and duration and microwave power for microwave blanching. The peppers treated by MWB, IRB, and HHAIB obtained higher red pigments and ascorbic acid retention, total antioxidant activity and DPPH values compared with the samples blanched by HWB. Ultrastructure analysis verified the difference phytochemical contents among four treatments. All four blanching treatments enhanced the drying kinetics relative to unblanched peppers. The Weibull distribution model was employed to calculate the moisture effective diffusivity ( D eff ), which ranged from 1.2688 × 10 −10 to 2.8754 × 10 −10 m 2 /s. The current findings not only contribute to a better understanding of different blanching methods but also provide more data and information for selecting blanching technologies. [ABSTRACT FROM AUTHOR]

Directly targeting caveolar caveolin-1 is a potential mechanism to regulate endothelial permeability, especially during oxidative stress, but little evidence on the topic limits therapeutics discoveries. In this study, we investigated the pharmacological effect of an antioxidant LM49 (5,2′-dibromo-2,4′,5′-trihydroxydiphenylmethanoe) and its five diphenylmethanone derivatives on endothelial permeability and establish two distinct mechanisms of action. Multiplex molecular assays with theoretical modeling indicate that diphenylmethanone molecules, including LM49, directly bind the caveolin-1 steric pocket of ASN53/ARG54, ILE49/ASP50, ILE18, LEU59, ASN60, GLU48 and ARG19 residues. They also indicated dynamic binding-affinity for diphenylmethanone derivatives. First, this molecular interaction at caveolin-1 pocket inhibits its phosphorylation at TYR14 residue in H 2 O 2 -injured endothelial cell. A positive correlation was established between diphenylmethanone derivative binding-affinity and caveolin-1 phosphorylation inhibition. Inhibition of caveolin-1 phosphorylation, however, was independent of the LM49-mediated variation of protein tyrosine kinase activity, suggesting a direct blockage of adenosine triphosphate substrate diffusion into cavelion-1 structure. Second, LM49 increases the expression of cellular adhesive and tight junction proteins, VE-cadherin and occludin, in H 2 O 2 -injured cell, in a dose dependent manner. A leakage assay of fluorescein isothiocyanate-labeled dextran 40 across cell monolayer suggested improvement in endothelial barrier integrity with diphenylmethanone treatments. Our results demonstrate a direct targeting effect of caveolin-1 on endothelial permeability, and should guide the diphenylmethanone therapy against oxidative stress-induced junction dysfunction, especially at caveolar membrane invagination. • Phosphor-CAV-1 shall be regarded as a marker of endothelial cell oxidative injury. • Such diphenylmethanones can directly bind to CAV-1 and inhibit its phosphorylation. • CAV-1 is a therapeutic target of LM49 to resist endothelial cell oxidative injury. • LM49 is a candidate to treat vascular disease related to endothelial permeability. [ABSTRACT FROM AUTHOR]