Our phosphoproteomics identified that phosphorylation of EphA2 at serine 897 (pS897-EphA2) was significantly upregulated in the high metastatic nasopharyngeal carcinoma (NPC) cells relative to non-metastatic NPC cells. However, the role and underlying mechanism of pS897-EphA2 in cancer metastasis and stem properties maintenance remain poorly understood. In this study, we established NPC cell lines with stable expression of exogenous EphA2 and EphA2-S897A using endogenous EphA2 knockdown cells, and observed that pS897-EphA2 maintained EphA2-dependent NPC cell in vitro migration and invasion, in vivo metastasis and cancer stem properties. Using phospho-kinase antibody array to identify signaling downstream of pS897-EphA2, we found that AKT/Stat3 signaling mediated pS897-EphA2-promoting NPC cell invasion, metastasis and stem properties, and Sox-2 and c-Myc were the effectors of pS897-EphA2. Immunohistochemistry showed that pS897-EphA2 was positively correlated with NPC metastasis and negatively correlated with patient overall survival. Moreover, ERK/RSK signaling controlled serum-induced pS897-EphA2 in NPC cells. Collectively, our results demonstrate that pS897-EphA2 is indispensable for EphA2-dependent NPC cell invasion, metastasis and stem properties by activating AKT/Stat3/Sox-2 and c-Myc signaling pathway, suggesting that pS897-EphA2 can serve as a therapeutic target in NPC and perhaps in other cancers.

Targeted delivery of antisense oligonucleotide (AON) drugs is a promising strategy to increase their concentration in the desired tissues and cell types while reducing access to other organs. Conjugation of AONs to N-acetylgalactosamine (GalNAc) has been shown to efficiently shift their biodistribution toward the liver via high-affinity binding to the asialoglycoprotein receptor (ASGPR) expressed at the surface of hepatocytes. Nevertheless, GalNAc conjugation does not prevent accumulation of AONs in the kidney cortex, and GalNAc-conjugated AONs might cause kidney toxicities, for example, under conditions of ASGPR saturation. Here, we investigated the nephrotoxicity potential of GalNAc-conjugated AONs by in vitro profiling of AON libraries in renal proximal tubule epithelial cells (PTECs) and in vivo testing of selected candidates. Whereas GalNAc-conjugated AONs appeared generally innocuous to PTECs, some caused mild-to-moderate nephrotoxicity in rats. Interestingly, the in vivo kidney liabilities could be recapitulated in vitro by treating PTECs with the unconjugated (or naked) parental AONs. An in vitro mechanistic study revealed that GalNAc conjugation attenuated AON-induced renal cell toxicity despite intracellular accumulation similar to that of naked AONs and independent of target knockdown. Overall, our in vitro findings reveal ASGPR-independent properties of GalNAc AONs that confer a favorable safety profile at the cellular level, which may variably translate in vivo due to catabolic transformation of circulating AONs.

Coronary reperfusion with HDL from healthy volunteers attenuates ischemia and reperfusion injury in animal models. In myocardial infarction (MI) patients, such an interaction is unclear. Hence, our first objective was to verify if there is interaction between HDL-C and MI mass in patients and the role of coronary reperfusion in the interaction. Furthermore, we investigated whether the effect in MI size of reperfusion with HDL obtained from healthy participants or MI patients could differ.

Alcohol use is associated with a variety of negative consequences, including heightened likelihood of cognitive impairment, proclivity to alcohol use disorders (AUD), and alterations in the drinker's offspring. Children and rodents exposed to alcohol during pregnancy, or those whose fathers consumed alcohol prior to mating, often exhibit neurodevelopmental, physiological, and behavioral deficits. The present study assessed cognitive function and alcohol intake in male and female rats that were offspring of alcohol-exposed fathers. Adult male rats were exposed to alcohol or vehicle (0.0 or 2.0 g/kg, respectively; twice daily for 2 days followed by a rest day, for a total of eight alcohol or vehicle exposure days), or were left untreated and then mated with non-manipulated females. The offspring were assessed for alcohol intake, via intraoral infusion, followed by cognitive assessment via an alternating T-maze task. The results indicated that paternal ethanol exposure, prior to breeding, resulted in offspring that consumed significantly more ethanol than vehicle or untreated controls. Furthermore, the offspring of alcohol-exposed fathers exhibited a significant failure to initiate and complete the T-maze performance tests. Although, when they did engage in the tests they performed at the level of controls (i.e., 80% correct). The present results add to a growing body of literature suggesting that paternal pre-conception alcohol exposure can have deleterious effects on the offspring.

There is evidence that training addicted participants to implicitly avoid disorder-related stimuli by using a training version of the Approach-Avoidance Task (AAT) results in reduced substance consumption (i.e., Approach Bias Modification [AppBM]). The aim of the present web-based study was to investigate the feasibility and effectiveness of AppBM in reducing gambling-related symptoms. A self-selected sample of participants with problem/pathological slot-machine gambling completed an online survey and received either AppBM or Sham training (final N = 131). Attrition during study participation was high (66%). In both conditions slot-machine related and neutral pictures were presented. Within the AppBM condition all slot-machine related pictures had to be pushed and all neutral pictures had to be pulled, whereas in the Sham condition the contingency was 50:50. Eight weeks after baseline, participants were re-assessed. Both groups showed a similar reduction in gambling-related symptoms. Findings are at odds with the hypothesis claiming that only contingency trainings yield beneficial effects. However, it cannot be ruled out that effects result from other factors unrelated to training such as expectancy effects. We think this study holds valuable information how to conduct larger trials in the future and may prove helpful to improve training and its delivery.

G-quadruplex (G4) structures have recently received increasing attention as a potential target for cancer research. We used time-gated fluorescence lifetime imaging microscopy (FLIM) with a G4 fluorescent probe, 3,6-bis(1-methyl-2-vinylpyridinium) carbazole diiodide (o-BMVC), to measure the number of o-BMVC foci, which may represent G4 foci, in cells as a common signature to distinguish cancer cells from normal cells. Here, the decrease in the number of o-BMVC foci in the pretreatment of cancer cells with TMPyP4, BRACO-19 and BMVC4 suggested that they directly bind to G4s in cells. In contrast, the increase in the number of o-BMVC foci in the pretreatment of cells with PDS and Hoechst 33258 (H33258) suggested that they do not inhabit the binding site of o-BMVC to G4s in cells. After the H33258 was removed, the gradual decrease of H33258-induced G4 foci may be due to DNA repair. The purpose of this work is to introduce o-BMVC foci as an indicator not only to verify the direct binding of potential G4 ligands to G4 structures but also to examine the possible effect of some DNA binding ligands on DNA integrity by monitoring the number of G4 foci in cells.

Yellow rust, a widely known destructive wheat disease, affects wheat quality and causes large economic losses in wheat production. Hyperspectral remote sensing has shown potential for the detection of plant disease. This study aimed to analyze the spectral reflectance of the wheat canopy in the range of 350⁻1000 nm and to develop optimal spectral indices to detect yellow rust disease in wheat at different growth stages. The sensitive wavebands of healthy and infected wheat were located in the range 460⁻720 nm in the early-mid growth stage (from booting to anthesis), and in the ranges 568⁻709 nm and 725⁻1000 nm in the mid-late growth stage (from filling to milky ripeness), respectively. All possible three-band combinations over these sensitive wavebands were calculated as the forms of PRI (Photochemical Reflectance Index) and ARI (Anthocyanin Reflectance Index) at different growth stages and assessed to determine whether they could be used for estimating the severity of yellow rust disease. The optimal spectral index for estimating wheat infected by yellow rust disease was PRI (570, 525, 705) during the early-mid growth stage with R² of 0.669, and ARI (860, 790, 750) during the mid-late growth stage with R² of 0.888. Comparison of the proposed spectral indices with previously reported vegetation indices were able to satisfactorily discriminate wheat yellow rust. The classification accuracy for PRI (570, 525, 705) was 80.6% and the kappa coefficient was 0.61 in early-mid growth stage, and the classification accuracy for ARI (860, 790, 750) was 91.9% and the kappa coefficient was 0.75 in mid-late growth stage. The classification accuracy of the two indices reached 84.1% and 93.2% in the early-mid and mid-late growth stages in the validated dataset, respectively. We conclude that the three-band spectral indices PRI (570, 525, 705) and ARI (860, 790, 750) are optimal for monitoring yellow rust infection in these two growth stages, respectively. Our method is expected to provide a technical basis for wheat disease detection and prevention in the early-mid growth stage, and the estimation of yield losses in the mid-late growth stage.

Glioblastoma multiform (GBM) tumors are very heterogeneous, organized in a hierarchical pattern, including cancer stem cells (CSC), and are responsible for development, maintenance, and cancer relapse. Therefore, it is relevant to establish new GBM cell lines with CSC characteristics to develop new treatments. A new human GBM cell line, named R2J, was established from the cerebro-spinal fluid (CSF) of a patient affected by GBM with leptomeningeal metastasis. R2J cells exhibits an abnormal karyotype and form self-renewable spheres in a serum-free medium. Original tumor, R2J, cultured in monolayer (2D) and in spheres showed a persistence expression of CD44, CD56 (except in monolayer), EGFR, Ki67, Nestin, and vimentin. The R2J cell line is tumorigenic and possesses CSC properties. We tested in vitro the anticancer effects of sodium selenite (SS) compared to temozolomide TMZ. SS was absorbed by R2J cells, was cytotoxic, induced an oxidative stress, and arrested cell growth in G2M before inducing both necrosis and apoptosis via caspase-3. SS also modified dimethyl-histone-3-lysine-9 (H3K9m2) levels and decreased histone deacetylase (HDAC) activity, suggesting anti-invasiveness potential. This study highlights the value of this new GBM cell line for preclinical modeling of clinically relevant, patient specific GBM and opens a therapeutic window to test SS to target resistant and recurrent GBM.

Ranking among the most lethal tumour entities, pancreatic duct adenocarcinoma cells invade neighbouring tissue resulting in high incidence of metastasis. They are supported by tumour stroma fibroblasts which have undergone differentiation into cancer-associated fibroblasts (CAFs). Stiffness of cell substratum, cytokines, such as transforming growth factor-β (TGF-β), and stromal matrix proteins, such as laminin-332, are factors which promote CAF differentiation. In a spheroid culture system, differentiation of CAFs was analysed for laminin-332 production, laminin-binding integrin repertoire, adhesion and migration behaviour, and, in heterospheroids, for their interplay with the pancreatic duct adenocarcinoma AsPC-I cells. Our data reveal that CAFs produce laminin-332 thus contributing to its ectopic deposition within the tumour stroma. Moreover, CAF differentiation correlates with an increased expression of α3β1 integrin, the principal laminin-332-receptor. Beyond its role as novel CAF marker protein, integrin α3β1 crucially determines differentiation and maintenance of the CAF phenotype, as knock-out of the integrin α3 subunit reversed the CAF differentiated state. AsPC-I cells co-cultured in heterospheroids with integrin α3-deficient CAFs invaded less than from heterospheroids with wild-type CAFs. This study highlights the role of integrin α3β1 integrin-laminin-332 interaction of CAFs which promotes and sustains differentiation of CAFs and promotes carcinoma invasion.

Inhibitors of chymase have good potential to provide a novel therapeutic approach for the treatment of cardiovascular diseases. We used a computational approach based on pharmacophore modeling, docking, and molecular dynamics simulations to evaluate the potential ability of 13 natural compounds from chamomile extracts to bind chymase enzyme. The results indicated that some chamomile compounds can bind to the active site of human chymase. In particular, chlorogenic acid had a predicted binding energy comparable or even better than that of some known chymase inhibitors, interacted stably with key amino acids in the chymase active site, and appeared to be more selective for chymase than other serine proteases. Therefore, chlorogenic acid is a promising starting point for developing new chymase inhibitors.

In this research survey the application of probiotic strain Lactobacillus plantarum ATCC 14917 in pomegranate juice fermentation is sought. Pomegranate juice was fermented for 24 h and then it was stored 4 for 4 weeks. Cell viability retained in high levels after the 24 h of fermentation and storage for 4 weeks (above 8.8 log cfu/mL), while fermented pomegranate juice was scored better at the 4th week of storage compared to non-fermented pomegranate juice. The probiotic strain was effective regarding lactic acid fermentation as was proved through sugar and organic acids analysis. Concentration of ethanol was maintained at low levels (0.3⁻1% v/v). Fermented pomegranate juice contained more and in higher percentages desirable volatile compounds (alcohols, ketones and esters) even at the 4th week of cold storage compared to non-fermented juice. Antioxidant activity (150.63 mg Trolox equivalent (TE)/100 mL at the 2nd week) and total phenolic content (206.46 mg gallic acid equivalents (GAE)/100 mL at the 2nd week) were recorded in higher levels for all the storage time compared to non-fermented juice.

A chemical-epigenetic method was used to enhance the chemodiversity of a marine algicolous fungus. Apart from thirteen known compounds, (+)-brevianamide R ((+)-3), (‒)-brevianamide R ((‒)-3), (+)-brevianamide Q ((+)-4), (‒)-brevianamide Q ((‒)-4), brevianamide V ((+)-5), brevianamide W ((‒)-5), brevianamide K (6), diorcinol B (7), diorcinol C (8), diorcinol E (9), diorcinol J (10), diorcinol (11), 4-methoxycarbonyldiorcinol (12), two new compounds, (+)- and (‒)-brevianamide X ((+)- and (‒)- 2)), as well as a new naturally occurring one, 3-[6-(2-methylpropyl)-2-oxo-1H-pyrazin-3-yl]propanamide (1), were isolated from chemical-epigenetic cultures of Aspergillus versicolor OUCMDZ-2738 with 10 µM vorinostat (SAHA). Compared to cultures in the same medium without SAHA, compounds 1⁻4, 8, 9, 11, and 12 were solely observed under SAHA condition. The structures of these compounds were elucidated based on spectroscopic analysis, specific rotation analysis, ECD, and X-ray crystallographic analysis. (±)-3, (±)-4, and (±)-5 were further resolved into the corresponding optically pure enantiomers and their absolute configurations were determined for the first time. Compounds 11 and 12 showed selective antibacterial against Pseudomonas aeruginosa with a minimum inhibitory concentration (MIC) of 17.4 and 13.9 μM, respectively. Compound 10 exhibited better α-glucosidase inhibitory activity than the assay control acarbose with IC50 values of 117.3 and 255.3 μM, respectively.

With the aim to obtain a site-specific doxorubicin (DOX) delivery in neuroblastoma SH-SY5Y cells, we designed an hybrid nanocarrier combining graphene oxide (GO) and magnetic iron oxide nanoparticles (MNPs), acting as core elements, and a curcumin⁻human serum albumin conjugate as functional coating. The nanohybrid, synthesized by redox reaction between the MNPs@GO system and albumin bioconjugate, consisted of MNPs@GO nanosheets homogeneously coated by the bioconjugate as verified by SEM investigations. Drug release experiments showed a pH-responsive behavior with higher release amounts in acidic (45% at pH 5.0) vs. neutral (28% at pH 7.4) environments. Cell internalization studies proved the presence of nanohybrid inside SH-SY5Y cytoplasm. The improved efficacy obtained in viability assays is given by the synergy of functional coating and MNPs constituting the nanohybrids: while curcumin moieties were able to keep low DOX cytotoxicity levels (at concentrations of 0.44⁻0.88 µM), the presence of MNPs allowed remote actuation on the nanohybrid by a magnetic field, increasing the dose delivered at the target site.

One fifth of U.S. adults have a current mental illness. Nutrition, physical activity, and sleep are critical to physical health; any related deficiencies may worsen existing mental health conditions. Little is known about the impact of clinical pharmacist assessment and consultation in improving physical and mental health outcomes. The study objective was to determine whether patients' mental health status improved following clinical pharmacist consultation. This pilot study involved clinical pharmacist-delivered services at an integrated medical behavioral health clinic in June 2018. Inclusion criteria required adults aged 18 years older, an established mental health diagnosis, and taking ≥2 prescribed psychotropic medications. One pharmacist conducted telephonic, medical, and psychiatric health risk assessment and counseling to improve nutrition, physical activity, and sleep status, both initially and at two-week follow-up. The Duke Health Profile (Duke) physical, anxiety, depression, and anxiety-depression scores measured patients' pre/post changes. Participants (n = 20) experienced higher Duke physical scores (p = 0.007) and significantly lower anxiety (p = 0.025), depression (p = 0.001) and anxiety-depression scores (p = 0.005) at follow-up. This pilot study provides preliminary evidence for pharmacist-led, targeted, telephonic counseling in improving short-term physical and mental Duke health scores. Further research evaluating the impact of clinical pharmacists' role in improving physical and behavioral health outcomes is warranted.

Threatened species recovery programmes benefit from incorporating genomic data into conservation management strategies to enhance species recovery. However, a lack of readily available genomic resources, including conspecific reference genomes, often limits the inclusion of genomic data. Here, we investigate the utility of closely related high-quality reference genomes for single nucleotide polymorphism (SNP) discovery using the critically endangered kakī/black stilt (Himantopus novaezelandiae) and four Charadriiform reference genomes as proof of concept. We compare diversity estimates (i.e., nucleotide diversity, individual heterozygosity, and relatedness) based on kakī SNPs discovered from genotyping-by-sequencing and whole genome resequencing reads mapped to conordinal (killdeer, Charadrius vociferus), confamilial (pied avocet, Recurvirostra avosetta), congeneric (pied stilt, Himantopus himantopus) and conspecific reference genomes. Results indicate that diversity estimates calculated from SNPs discovered using closely related reference genomes correlate significantly with estimates calculated from SNPs discovered using a conspecific genome. Congeneric and confamilial references provide higher correlations and more similar measures of nucleotide diversity, individual heterozygosity, and relatedness. While conspecific genomes may be necessary to address other questions in conservation, SNP discovery using high-quality reference genomes of closely related species is a cost-effective approach for estimating diversity measures in threatened species.

When sows are mixed into groups, hierarchies form and resulting aggression and stress can affect production and welfare. This study determined the effect of providing point-source materials on aggressive and play behaviors in gestating sows. Large white cross Landrace sows were mixed after insemination; six pens of 12 sows were housed in 'standard' pens, and six pens of 12 sows were housed in 'enhanced' pens. The 'enhanced' pens each contained two rubber mats, eight strands of 24 mm-thick sisal rope and two yellow plastic disks, suspended from the roof. The sows remained in these pens until pregnancy confirmation. Salivary cortisol concentration, injury counts, and sow behaviors were recorded the day before mixing (day 1), mixing (day 0) and post-mixing day 1, day 4, day 7 and day 20. At farrowing, reproductive outcomes were obtained. Play was observed (including locomotor and object play) in the 'enhanced' pen, and percentage of time spent playing was greater on d4 (1.48 ± 0.3 Square root transformed data (2.84% non-transformed adjusted mean)), d7 (1.43 ± 0.3 (2.97%)) and d20 (1.64 ± 0.3 (3.84%)), compared to d0 (0.56 ± 0.3 (0.70%)) and d1 (0.87 ± 0.3 (1.67%) (p < 0.05)). No play was observed in standard housing. Aggression, salivary free cortisol concentrations and injuries were unaffected (p > 0.05). The provision of materials had no impact on aggression, although their presence maintained sow interest and play behavior, suggesting a positive effect.

Tandem mass spectral databases are indispensable for fast and reliable compound identification in nontargeted analysis with liquid chromatography⁻high resolution tandem mass spectrometry (LC-HRMS/MS), which is applied to a wide range of scientific fields. While many articles now review and compare spectral libraries, in this manuscript we investigate two high-quality and specialized collections from our respective institutes, recorded on different instruments (quadrupole time-of-flight or QqTOF vs. Orbitrap). The optimal range of collision energies for spectral comparison was evaluated using 233 overlapping compounds between the two libraries, revealing that spectra in the range of CE 20⁻50 eV on the QqTOF and 30⁻60 nominal collision energy units on the Orbitrap provided optimal matching results for these libraries. Applications to complex samples from the respective institutes revealed that the libraries, combined with a simple data mining approach to retrieve all spectra with precursor and fragment information, could confirm many validated target identifications and yield several new Level 2a (spectral match) identifications. While the results presented are not surprising in many ways, this article adds new results to the debate on the comparability of Orbitrap and QqTOF data and the application of spectral libraries to yield rapid and high-confidence tentative identifications in complex human and environmental samples.

Dengue virus (DENV) is one of the most common arbovirus diseases, with a wide spectrum of presentation. Spinal cord involvement in dengue infection (DF) is rare. However, the risk of transverse myelitis (TM) following Dengue has not been systematically assessed.

The emergence of multidrug-resistant tuberculosis (MDR⁻TB) and more recently, extensively drug-resistant (XDR) TB has intensified the need for studies aimed at identifying factors associated with TB drug resistance. This study determined the differences in plasma concentrations of pro-inflammatory (IFN-γ and TNF-α) and anti-inflammatory (IL-10) cytokines in MDR-TB and drug-susceptible (DS) TB patients, in addition to some socio-economic factors. Plasma levels of IL-10, IFN-γ and TNF-α were measured in 83 participants (comprising 49 MDR-TB and 34 DS-TB patients) using sandwich ELISA. Levels of the three cytokines were elevated in MDR-TB patients compared to DS-TB patients. The mean level of IL-10 (7.8 ± 3.61 ρg/mL) measured in MDR-TB cases was relatively higher than those of TNF-α and IFN-γ, and statistically significant (p = 0.0022) when compared to the level of IL-10 (4.8 ± 4.94 ρg/mL) in the DS-TB cases. There were statistically significant associations between MDR-TB and factors such as education level (X² = 9.895, p = 0.043), employment status (X² = 19.404, p = 0.001) and alcoholism (X² = 3.971, p = 0.046). This study adds to the knowledge that IFN-γ, TNF-α and IL-10 play a role in the host response to Mycobacterium tuberculosis (MTB). Alcohol intake can be considered as an important MDR-TB risk factor.

On the tenth anniversary of the 2008 Wenchuan Earthquake, investigating the evolution of disaster science is worthwhile and can be used to improve the future execution of disaster risk management. Based on more than 55,786 articles on the relative topic of "Disaster" derived from the Web of Science Core Collection from 1999⁻2017, this study employs CiteSpace and Google Earth to identify and visualize the spatial distribution of publications, bursts of keywords and categories, highly cited references, and interdisciplinary levels and then identify the emerging trends of disaster research over the past 20 years. The results show that the earthquake indeed jumpstarted a massive wave of disaster research around the world and increased international cooperation over the last decade. However, in terms of both the quantity and quality of publications in disaster research fields, China is lagging behind the U.S. and European countries. Moreover, although designing disaster prevention and mitigation strategies is a new popular field of disaster science, geological environment changes and geologic hazards triggered by earthquakes are more popular research topics than disaster emergency and recovery. In addition, the transdisciplinary level of disaster science increased after the earthquake. This interdisciplinary characteristic of disaster science gradually increased in popularity, which demonstrates that people can learn from catastrophes. These emerging trends could serve as a scientific basis to clearly understand disaster science progress over the last 20 years and provide a reference for rapidly identifying frontier issues in disaster science.