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To investigate the pivotal roles of the duck plague virus (DPV) tegument protein UL14 in viral replication, we generated 2 mutated viruses of DPV by using the bacterial artifcial chromosome system, the UL14-null mutant virus (CHv-BAC-ΔUL14) and the corresponding revertant virus (CHv-BAC-ΔUL14R). We found that the CHv-BAC-ΔUL14 viruses exhibited impaired virion morphogenesis in transmission electron microscopy (TEM) studies. Furthermore, CHv-BAC-ΔUL14 exhibited a plaque size reduction in duck embryo fibroblasts (DEFs). Finally, CHv-BAC-ΔUL14 exhibited a significant viral growth defect. Taken together, our findings suggest that DPV UL14 protein regulates viral morphogenesis for efficient viral replication. [ABSTRACT FROM AUTHOR]

Alternative cleavage and polyadenylation (APA) results in mRNA isoforms containing different 3’ untranslated regions (3’UTRs) and/or coding sequences. How core cleavage/polyadenylation (C/P) factors regulate APA is not well understood. Using siRNA knockdown coupled with deep sequencing, we found that several C/P factors can play significant roles in 3’UTR-APA. Whereas Pcf11 and Fip1 enhance usage of proximal poly(A) sites (pAs), CFI-25/68, PABPN1 and PABPC1 promote usage of distal pAs. Strong cis element biases were found for pAs regulated by CFI-25/68 or Fip1, and the distance between pAs plays an important role in APA regulation. In addition, intronic pAs are substantially regulated by splicing factors, with U1 mostly inhibiting C/P events in introns near the 5’ end of gene and U2 suppressing those in introns with features for efficient splicing. Furthermore, PABPN1 inhibits expression of transcripts with pAs near the transcription start site (TSS), a property possibly related to its role in RNA degradation. Finally, we found that groups of APA events regulated by C/P factors are also modulated in cell differentiation and development with distinct trends. Together, our results support an APA code where an APA event in a given cellular context is regulated by a number of parameters, including relative location to the TSS, splicing context, distance between competing pAs, surrounding cis elements and concentrations of core C/P factors. [ABSTRACT FROM AUTHOR]

* Premise of the study: Bombax ceiba (Malvaceae), commonly known as silk cotton tree, is a multipurpose tree species of tropical forests. Novel expressed sequence tag-simple sequence repeat (EST-SSR) markers were developed and characterized for the species using transcriptome analysis. * Methods and Results: A total of 33 new EST-SSR markers were developed for B. ceiba, of which 13 showed polymorphisms across the 24 individuals from four distant populations tested in the study. The results showed that the number of alleles per polymorphic locus ranged from two to four, and the expected heterozygosity and observed heterozygosity per locus varied from 0.043 to 0.654 and from 0 to 0.609, respectively. * Conclusions: These newly developed EST-SSR markers can be used in phylogeographic and population genetic studies to investigate the origin of B. ceiba populations. Furthermore, these EST-SSR markers could also greatly promote the development of molecular breeding studies pertaining to silk cotton tree. [ABSTRACT FROM AUTHOR]

The purpose of this work was to explore the feasibility of using Soluplus ® in preparing a fenofibrate (FBT) nanosuspension adopting wet media milling technology. HPMC and Soluplus ® were used as stabilizers to prepare FBT/HPMC nanosuspension (F1) and FBT/Soluplus ® nanosuspension (F2), respectively. The nanosuspensions were subjected to evaluations involving particle size, dissolution, preliminary stability and pharmacokinetic behavior. A marked reduction in particle size was achieved by nanosuspensions (from 17.55 μm to 642 nm (F1) and 344 nm (F2)). The nanosuspensions displayed almost complete dissolution while percentages of 30% and 13% were obtained by physical mixtures and coarse FBT separately. Soluplus ® could stabilize the nanosuspension more effectively due to a weaker Ostwald ripening effect resulting from a slower diffusion of micelles formed by Soluplus ® entrapping dissolved FBT than FBT exposed to pure water directly. In the in vivo evaluation, larger AUC 0–72 h and C max , and shorter T max were obtained by the nanosuspensions. Significant differences were observed between the physical mixtures. The phenomenon of double peaks was present in this study. The major factor may be the multiple absorption sites of FBT. The current work indicated that Soluplus ® is well suited for preparation of a nanosuspension with good stability and improved dissolution and bioavailability. [ABSTRACT FROM AUTHOR]

[Display omitted] • Deep penetrating sealer (DPS) for concrete superficial protection. • The DPS improve the compactness and the water impermeability of the concrete. • Sodium silicate in DPS and C–H in concrete contributes to the increase of C-S-H. • DPS seal the pores with pore size ranging from 20 nm to 130 nm inside the concrete. • The quantity of newly generated C-S-H is higher in the concrete cracks and pores. Deep penetrating sealer (DPS) is water-based permeable crystalline waterproofing coating, and it is widely used to improve the impermeability and durability of cement-based materials. This study investigated the action mechanism of DPS on cement-based materials, especially the microstructure of the newly generated substances and pore evolution in concrete that DPS treated. To explore the action mechanism and permeability of DPS on cement-based materials, experimental specimens were fabricated in this experiment. The specimens included concrete, cement paste, dried DPS, and the product reacted by DPS mixed with saturated calcium hydroxide solution. To test the microstructure, chemical composition, pore structure, and macroscopic permeability of the specimens, scanning electron microscopy, x-ray diffraction, energy dispersive spectrometer, nitrogen absorption, and AutoClam equipment were employed. The experimental and analytical results showed that the calcium silicate hydrate content was increased by the chemical reaction between DPS and cement-based materials. The microstructure of concrete surface then became more densified. The volume of pore with a pore size ranging from 20 to 130 nm inside concrete was decreased, and the water penetration of the concrete was reduced by approximately 75%. This study revealed the action mechanism of DPS and cement-based materials, providing reference for the improvement of DPS and practical engineering application. [ABSTRACT FROM AUTHOR]

Soy protein isolate (SPI) is often used in food processing due to the high water holding capacity and high stability. This paper explored the effect of heat treatment (HT) on the nonlinear rheological properties of acid-Induced soy protein isolate gels modified by high-pressure homogenization. The effects of HT on the storage modulus G′ , loss modulus G″ and gel rate of the SPI gel were studied through time sweep, frequency sweep and strain sweep tests. The effects of the Lissajous curve, Chebyshev stress decomposition and Fourier transformation were also investigated. The results illustrated that the HT significantly enhanced the mechanical properties of the SPI gel. The curve area and energy dissipation of the Lissajous curve after HT were significantly reduced. Chebyshev stress decomposition illustrated that HT enhanced the shear thickening behavior of SPI gel. The study also found that HT significantly improved the structure and strain resistance of the SPI gel structure. In short, HT significantly improved the nonlinear rheological properties of the SPI gel. • Gel network structure of HIPPE can be stabilized by A/S-SPIG particles. • SPI particles can improve the stability of HIPPE at low temperatures. • A/S-SPIG particle concentration is positively correlated with the emulsification stability. [ABSTRACT FROM AUTHOR]

The effect of catalyst height (0–50 mm) and catalyst temperature (600–900 °C) on the catalytic cracking of gaseous tar is investigated using a two-stage down reactor. Results show that the utilization of catalyst promotes the removal of coal tar. The yield of coal tar at the pyrolysis temperature of 900 °C is 7.14% and tar is composed of aromatics and high-carbon-number aliphatics. The coal char prepared by fast coke-making method at 900 °C is directly used as a catalyst in the catalytic cracking of coal tar. When the catalyst temperature and the catalyst height are 900 °C and 50 mm, the tar produced by coal pyrolysis is connected with high-temperature coal char for a short time, reducing the tar yield from 7.14% to 4.26%. Among them, aliphatic hydrocarbons with carbon number of C10 and C11 and aromatics are the main components of coal tar. Similarly, compared with the pyrolysis, the content of CH 4 and H 2 obtained with the addition of the catalyst increase from 0.56 to 0.97 to 1.64%g/g coal and 1.02%g/g coal, respectively. Moreover, GC/MS results show that the increment of catalyst height and catalyst temperature increases aromatics content and facilitates the conversion of high-carbon-number component to low-carbon-number ones. • The char produced by coal pyrolysis is directly used as a catalyst in the catalytic cracking of gaseous tar. • The short-time contact between tar with high-temperature coal char reduces the yield of tar. • The yield and composition of gas and tar with the addition of char change significantly. • Attempt to propose the mechanism of tar catalytic cracking process over char catalysts. [ABSTRACT FROM AUTHOR]

[Display omitted] • The difference in thermal conductivity between VCT and SCT is obtained. • Effect mechanism of VCT is explained via porosity and microstructure evolutions. • The low temperature in VCT has a greater effect on hydration than the high temperature. • Prediction model of concrete thermal conductivity under VCT condition is built. The thermal conductivity and microevolution mechansim of early-age concrete (EAC) under variable curing temperature environments significantly influence the temperature cracks in massive concrete. An experiment was carried out to compare thermal conductivity evolution differences of EAC under variable curing temperature (VCT) and standard curing temperature (SCT) conditions. The concrete specimens of moisture and dry states were used to consider the effect of moisture content on thermal conductivity. Results showed that the moisture concrete thermal conductivity under VCT condition decreases with curing ages and is higher than SCT condition. For the dry samples, the thermal conductivity of VCT condition increases with curing ages and is lower than the SCT condition. The difference in thermal conductivity between VCT and SCT increases from zero to seven days and decreases after seven days. The scanning electron microscope (SEM) tests showed that the VCT leads to slower growth of pore structure and larger porosity of EAC due to the slower hydration rate under VCT condition. The thermal conductivity difference between VCT and SCT is mainly caused by the differences in water content, chemical components in solid phase, and porosity (solid-phase volume fraction). Meanwhile, the relation among thermal conductivity, porosity and most probable pore size shows that curing temperature has a great effect on hydration degree and controls the thermal conductivity evolution. Besides, a prediction model of EAC thermal conductivity under VCT condition was built based on hydration kinetics. [ABSTRACT FROM AUTHOR]

• A new method of I-ELISA based on the DHAV-1 VP4 protein was established. • The VP4 gene is located inside the viral capsid, but the antibodies it produces have neutralizing activity. • VP4 protein has the advantages of low variability. • The method is another option for monitoring DHAV-1 infection or prevalence. In Picornavirus, the VP4 gene is located inside the viral capsid, but the antibodies it produces have neutralizing activity. At the same time, we know that the VP4 gene is relatively conserved among the four structural proteins, which makes the usage VP4 protein-based antigen potentially meaningful. Therefore, we used purified duck hepatitis A virus type 1 (DHVA-1) recombinant VP4 protein as the coating antigen to establish an indirect method. The optimal antigen, serum and enzyme-labeled antibody dilutions were 1:400 (3.375 μg/mL), 1:80 and 1:1600, respectively. The optimal blocking buffer was 1% skimmed milk powder. The cutoff value was determined to be 0.203, and the analytical sensitivity was 1:1600. The established ELISA method has good specificity, repeatability and sensitivity. It has a high coincidence rate of 70.8 % with the DHVA-1 as the coating antigen. So, it can be used for the detection of DHAV-1 serum antibodies. [ABSTRACT FROM AUTHOR]

Despite the well-known effects that Quaternary climate oscillations had on shaping intraspecific diversity, their role in driving homoploid hybrid speciation is less clear. Here, we examine their importance in the putative homoploid hybrid origin and evolution of Ostryopsis intermedia, a diploid species occurring in the Qinghai-Tibet Plateau ( QTP), a biodiversity hotspot. We investigated interspecific relationships between this species and its only other congeners, O. davidiana and O. nobilis, based on four sets of nuclear and chloroplast population genetic data and tested alternative speciation hypotheses. All nuclear data distinguished the three species clearly and supported a close relationship between O. intermedia and the disjunctly distributed O. davidiana. Chloroplast DNA sequence variation identified two tentative lineages, which distinguished O. intermedia from O. davidiana; however, both were present in O. nobilis. Admixture analyses of genetic polymorphisms at 20 SSR loci and sequence variation at 11 nuclear loci and approximate Bayesian computation ( ABC) tests supported the hypothesis that O. intermedia originated by homoploid hybrid speciation from O. davidiana and O. nobilis. We further estimated that O. davidiana and O. nobilis diverged 6-11 Ma, while O. intermedia originated 0.5-1.2 Ma when O. davidiana is believed to have migrated southward, contacted and hybridized with O. nobilis possibly during the largest Quaternary glaciation that occurred in this region. Our findings highlight the importance of Quaternary climate change in the QTP in causing hybrid speciation in this important biodiversity hotspot. [ABSTRACT FROM AUTHOR]

Abstract: In the paper, we begin by describing polyethylene glycol–poly lactic acid-co-glycolic acid (PEG–PLGA) which was chosen as a typical model copolymer for the construction of nano-sized drug delivery systems and also the types of PEG–PLGA copolymers that were eluted. Following this we examine the structure-influenced drug delivery applications including nanoparticles, micelles and hydrogels. After that, the preparation methods for nano-sized delivery systems are presented. In addition, the drug loading mode of PEG–PLGA micelles is divided into three aspects. Finally, the drug release profiles of PEG–PLGA micelles, both in terms of their in vitro and in vivo characteristics, are represented. PEG–PLGA copolymers are very suitable for the construction of micelles as carriers for insoluble drugs. This article reviews the structure and the different structure-influenced applications of PEG–PLGA copolymers, concentrating on the application of PEG–PLGA micelles. [Copyright &y& Elsevier]

Intravenous lipid emulsions of cabazitaxel (CLEs) with a high stability were prepared by adding cholesterol (CH) to provide a new and more suitable delivery system for its administration. The factors affecting CLEs, such as the solubility of cabazitaxel in various oils, different kinds of lecithin, pH, different types of oil phases, and different concentrations of lipoid E80®, CH and poloxamer 188 were investigated systematically. The degradation of cabazitaxel in aqueous solution and lipid emulsion both followed pseudo first-order kinetics. A degradation mechanism was suggested by the U-shaped pH-rate profile of cabazitaxel. A formulation containing 0.5% (w/v) CH and another formulation without CH were made to investigate the protective influence of CH on the chemical stability of CLEs. The activation energy of the two formulations was calculated to be 65.74±6.88 and 54.24±1.43kJ/mol (n =3), respectively. Compared with the untreated CH, the shelf-life of cabazitaxel with added CH was longer, namely 134.0±23.4days versus 831.4±204.4days (n =3) at 4°C. This indicates that the addition of CH significantly improved the lifetime of cabazitaxel in intravenous lipid emulsions. The hydrogen bonding that takes place between cabazitaxel and CH accounts for the protective effect of CH on the chemical stability of CLEs in two ways: preventing cabazitaxel from leaking and hydrolyzing in aqueous solution and hindering hydrolysis in the oil phase. Finally, the hypothesis was confirmed by LC/TOFMS and Fourier-transform infrared-spectroscopy. As a result, CLEs were obtained successfully by the addition of CH and were stable enough to allow further research. [ABSTRACT FROM AUTHOR]

Abstract: The purpose of this article was to compare the in vitro and in vivo profiles of itraconazole (ITZ) extrudates and nanosuspension separately prepared by two different methods. And it was proved truly to form nanocrystalline and amorphous ITZ characterized by differential scanning calorimetry (DSC), X-ray powder diffraction (XRD) analysis, Fourier transform infrared spectrum (FTIR), transmission electron microscope (TEM), and scanning electron microscope (SEM). The release of ITZ/Soluplus solid dispersions with amorphous ITZ was almost complete while only 40% release was obtained with ITZ nanocrystals. The amorphous state need not to cross over the crystal lattice energy upon dissolution while the crystalline need to overcome it. In the in vivo assay, the AUC(0–t) and C max of ITZ/Soluplus were 6.9- and 11.6-time higher than those of pure ITZ. The formulation of the extrudate had an AUC(0–t) and C max similar to those of ITZ and also OH-ITZ compared with the commercial capsule (Sporanox®). The relative bioavailability values with their 95% confidence limit were calculated to be 98.3% (92.5–104.1%) and 101.3% (97.9–104.1%), respectively. The results of this study showed increased dissolution and bioavailability of the solid dispersion of Soluplus-based carrier loading ITZ prepared by HME compared with the ITZ nanosuspension prepared by wet milling. [Copyright &y& Elsevier]

• DHAV-1 2C protein is an NTPase. • The ATPase activity of DHAV-1 2C protein was inhibited by guanidine hydrochloride (GdnHCl). • The lysine at position 151 of the duck hepatitis A virus 1 2C protein is critical for its NTPase activities. The duck hepatitis A virus 1 (DHAV-1) 2C protein was predicted to be a superfamily III helicase member and includes nucleotide binding (NTB) and putative RNA helicase activity motifs. To study whether DHAV-1 2C protein has NTB activity, we expressed DHAV-1 2C protein with maltose binding protein (MBP) to solve its poor solubility in a prokaryotic expression system. We showed that the DHAV-1 2C protein has nucleoside triphosphatase (NTPase) activity by measuring the released phosphate. The NTPase of the DHAV-1 2C protein is Mg2+ indispensable and affected by other biochemical characteristics such as Mn2+, Ca2+, Zn2+, Na+ and pH. Guanidine hydrochloride (GdnHCl), a potent inhibitor of viral RNA replication, inhibited ATPase activity of the DHAV-1 2C protein in a dose-dependent manner. Finally, we constructed three mutants to identify the key site for the ATPase activity of the DHAV-1 2C protein. These results indicate that lysine at position 151 of the DHAV-1 2C protein is very important for NTPase activity. Here, we demonstrated and partially characterized that the DHAV-1 2C protein has NTPase activity and showed that mutation of the lysine in the conserved Walker A impairs that activity. The results serve to confirm what is readily predicted from previous work on picornavirus 2C proteins. It also provides a basis for further study of the 2C protein and the function of NTPase activity on the viral life cycle. [ABSTRACT FROM AUTHOR]

• TMUV NS5 contains a suboptimal NLS and does not appreciably accumulate in the nucleus. • Two arginine mutations N390R and Q392R of the NS5 bipartite nuclear localization sequence (α/βNLS) are sufficient to transport NS5 into the nucleus. • Nuclear localization of NS5 strongly reduces virus replication and production in vitro but exhibits no obvious pathogenicity in vivo. • A small subset of NS5 NLSmut showed to interact with NS3 under transfection and infection conditions. • NS5 NLSmut didn't not compromise interferon production mediated by RIG-I detection of the viral RNA polymerase activity. Duck Tembusu virus (TMUV) belongs to the flavivirus genus whose genome replication involved in capping and RNA synthesis dominating by nonstructural protein 5 (NS5). Flaviviral replication has been well documented to occur in the cytoplasm, but the effect of NS5 to gain access to the nucleus remains controversial. Here, TMUV NS5 was observed to localize within the cytoplasm of transfected and infected cells and co-localized with the endoplasmic reticulum. We introduced two arginine mutations into the N390 and Q392 (N390R and Q392R) of the NS5 bipartite nuclear localization sequence (α/βNLS) and designated that mutagenesis as NS5 NLSmut , which has shown the ability to access the nucleus and hence attenuates viral replication and production in vitro. Additionally, there was no significant difference between the recovered wild-type TMUV (rTMUV-WT) and engineered mutant (rTMUV-NS5 NLSmut) on plaque morphology, survival rate of infected duck embryos or virus copies in tissues. Considering that NS5 NLSmut is mainly located in the cytoplasm of rTMUV-NS5 NLSmut infected cells at the early stage of infection. We further confirmed that NS5 NLSmut attenuated its interaction with nonstructural NS2B-NS3 (NS2B3) following transfection and infection. Meanwhile, the rTMUV-NS5 NLSmut tended to stimulate more interferon beta (IFNβ) than rTMUV-WT. However, preliminary study on transient NS5 and NS5 NLSmut detected the same levels of IFNβ mRNA mediated by RIG-I detection of NS5 RNA polymerase activity in cell. In summary, these results provide further insights into the relationship between the viral property and subcellular localization of flavivirus NS5 in terms of the NS5-NS2B3 interaction. [ABSTRACT FROM AUTHOR]

• One deeply represented multiple instance learning (DR-MIL) method is proposed. • DR-MIL can distinguish COVID-19 from community acquired pneumonia via CT images. • DR-MIL is a weak-supervised learning and applicable to the small dataset. • The generated Deep Represented Instance Score is an imaging biomarker of COVID-19. • The spatial pattern of infected lesions in COVID-19 is different from that of CAP. Given that the novel coronavirus disease 2019 (COVID-19) has become a pandemic, a method to accurately distinguish COVID-19 from community-acquired pneumonia (CAP) is urgently needed. However, the spatial uncertainty and morphological diversity of COVID-19 lesions in the lungs, and subtle differences with respect to CAP, make differential diagnosis non-trivial. We propose a deep represented multiple instance learning (DR-MIL) method to fulfill this task. A 3D volumetric CT scan of one patient is treated as one bag and ten CT slices are selected as the initial instances. For each instance, deep features are extracted from the pre-trained ResNet-50 with fine-tuning and represented as one deep represented instance score (DRIS). Each bag with a DRIS for each initial instance is then input into a citation k -nearest neighbor search to generate the final prediction. A total of 141 COVID-19 and 100 CAP CT scans were used. The performance of DR-MIL is compared with other potential strategies and state-of-the-art models. DR-MIL displayed an accuracy of 95% and an area under curve of 0.943, which were superior to those observed for comparable methods. COVID-19 and CAP exhibited significant differences in both the DRIS and the spatial pattern of lesions (p <0.001). As a means of content-based image retrieval, DR-MIL can identify images used as key instances, references, and citers for visual interpretation. DR-MIL can effectively represent the deep characteristics of COVID-19 lesions in CT images and accurately distinguish COVID-19 from CAP in a weakly supervised manner. The resulting DRIS is a useful supplement to visual interpretation of the spatial pattern of lesions when screening for COVID-19. [ABSTRACT FROM AUTHOR]

The human gene encoding the cleavage/polyadenylation (C/P) factor CstF-77 contains 21 exons. However, intron 3 (In3) accounts for nearly half of the gene region, and contains a C/P site (pA) with medium strength, leading to short mRNA isoforms with no apparent protein products. This intron contains a weak 5′ splice site (5′SS), opposite to the general trend for large introns in the human genome. Importantly, the intron size and strengths of 5′SS and pA are all highly conserved across vertebrates, and perturbation of these parameters drastically alters intronic C/P. We found that the usage of In3 pA is responsive to the expression level of CstF-77 as well as several other C/P factors, indicating it attenuates the expression of CstF-77 via a negative feedback mechanism. Significantly, intronic C/P of CstF-77 pre-mRNA correlates with global 3′UTR length across cells and tissues. In addition, inhibition of U1 snRNP also leads to regulation of the usage of In3 pA, suggesting that the C/P activity in the cell can be cross-regulated by splicing, leading to coordination between these two processes. Importantly, perturbation of CstF-77 expression leads to widespread alternative cleavage and polyadenylation (APA) and disturbance of cell proliferation and differentiation. Thus, the conserved intronic pA of the CstF-77 gene may function as a sensor for cellular C/P and splicing activities, controlling the homeostasis of CstF-77 and C/P activity and impacting cell proliferation and differentiation. [ABSTRACT FROM AUTHOR]

Abstract: PKKKRKV (Pro-Lys-Lys-Lys-Arg-Lys-Val, PV7), a seven amino acid peptide, has emerged as one of the primary nuclear localization signals that can be targeted into cell nucleus via the nuclear import machinery. Taking advantage of chemical diversity and biological activities of this short peptide sequence, in this study, Pluronic F127 nanomicelles engineered with nuclear localized functionality were successfully developed for intracellular drug delivery. These nanomicelles with the size ~100nm were self-assembled from F127 polymer that was flanked with two PV7 sequences at its both terminal ends. Hydrophobic anticancer drug doxorubicin (DOX) with inherent fluorescence was chosen as the model drug, which was found to be efficiently encapsulated into nanomicelles with the encapsulation efficiency at 72.68%. In comparison with the non-functionalized namomicelles, the microscopic observation reveals that PV7 functionalized nanomicelles display a higher cellular uptake, especially into the nucleus of HepG2 cells, due to the nuclear localization signal effects. Both cytotoxicity and apoptosis studies show that the DOX-loaded nanomicelles were more potent than drug nanomicelles without nuclear targeting functionality. It was thus concluded that PV7 functionalized nanomicelles could be a potentially alternative vehicle for nuclear targeting drug delivery. [Copyright &y& Elsevier]

Abstract: High levels of organochlorine pesticides (OCPs) such as hexachlorocyclohexanes (HCHs) and dichlorodiphenyltrichloroethanes (DDTs) have been found in soil of the Pearl River Delta (PRD), attributable to high pesticide application in this area. Consequently, the occurrence and environmental effect of HCHs and DDTs in the PRD have attracted considerable attention. However, study focusing on the influence of potential factors such as soil property on the environmental fate of HCHs and DDTs in the PRD has been rare. The present study aimed to investigate the impact of soil physiochemical properties on the distribution patterns and fate of soil HCHs and DDTs on a large spatial scale. Levels of HCHs (sum of α-, β-, γ- and δ-HCH) and DDTs (sum of 1,1,1-trichloro-2,2-bis-(p-chlorophenyl)ethane (p,p′-DDT), 1,1-dichloro-2,2-bis-(p-chlorophenyl)ethane (p,p′-DDD), and 1,1-dichloro-2,2-bis-(p-chlorophenyl)ethylene (p,p′-DDE)) in 151 soil samples covering all areas of the PRD and physiochemical parameters related to soil properties including pH, total organic carbon (TOC), total Fe (TFe), DCB-Fe (DFe), amorphous-Fe (AFe), complexed-Fe (CFe), total Mn (TMn), DCB-Mn (DMn), amorphous-Mn (AMn), complexed-Mn (CMn) and cation exchange capacity (CEC) were determined. The residual levels of HCHs and DDTs in soils of the present study, which are mainly controlled by soil TOC and CFe content and varying spatially with land use types, may potentially pose ecological risk to plants and animals. On the other hand, transformation of soil HCHs may be affected by pH and DDT transformation correlated significantly with AFe and CFe. Currently, soil has become an important secondary source of OCPs and the re-emission potential of OCPs in soil was mainly affected by soil OCP concentrations and land use types. [Copyright &y& Elsevier]

Abstract: The aim of this study was to develop an efficient method of preparing less irritant clarithromycin-loaded liposomes (CLA-Lip) for injection with a high drug loading and to evaluate their physicochemical characteristics before and after lyophilization. CLA-Lip were prepared using the film-dispersion method with sodium cholesterol sulfate (SCS) and n-hexyl acid as the regulators and then lyophilized. The liposomes were characterized in terms of their size, size distribution, zeta potential, morphology, in vitro release, haemolysis, and lyophilization and irritation testing was carried out. The TEM images revealed that the structure of the CLA-Lip were multilamellar and of a regular size of around 100nm. In addition, the lyophilized CLA-Lip were characterized by DSC and Infrared spectroscopy to confirm the structure. H-bonding and salt-forming reactions were used to ensure that clarithromycin (CLA) was stably encapsulated in the liposomes. This method provided a 30-fold increase in the concentration of clarithromycin relative to that in aqueous solution. Sucrose was found to be the best protective agent and was added in an amount of 12.5% (w/v). According to the mouse scratch test and the rat paw lick test, the pain of CLA-Lip was significantly reduce by approximately 80% compared with the solution of clarithromycin phosphate. In addition, rabbit ear vein experiments produced similar results. These findings suggested that CLA-Lip was a stable delivery system with less irritation, which should be extremely suitable for clinical application. [Copyright &y& Elsevier]

Abstract: Recently we described an ischemic preconditioning induced by repetitive coronary stenosis, which is induced by 6 episodes of non-lethal ischemia over 3days, and which also resembles the hibernating myocardium phenotype. When compared with traditional second window of ischemic preconditioning using cDNA microarrays, many genes which differed in the repetitive coronary stenosis appeared targeted to metabolism. Accordingly, the goal of this study was to provide a more in depth analysis of changes in metabolism in the different models of delayed preconditioning, i.e., second window and repetitive coronary stenosis. This was accomplished using a metabolomic approach based on liquid chromatography–mass spectrometry (LC–MS) and gas chromatography–mass spectrometry (GC–MS) techniques. Myocardial samples from the ischemic section of porcine hearts subjected to both models of late preconditioning were compared against sham controls. Interestingly, although both models involve delayed preconditioning, their metabolic signatures were radically different; of the total number of metabolites that changed in both models (135 metabolites) only 7 changed in both models, and significantly more, p <0.01, were altered in the repetitive coronary stenosis (40%) than in the second window (8.1%). The most significant changes observed were in energy metabolism, e.g., phosphocreatine was increased 4 fold and creatine kinase activity increased by 27.2%, a pattern opposite from heart failure, suggesting that the repetitive coronary stenosis and potentially hibernating myocardium have enhanced stress resistance capabilities. The improved energy metabolism could also be a key mechanism contributing to the cardioprotection observed in the repetitive coronary stenosis and in hibernating myocardium. This article is part of a Special Issue entitled “Focus on Cardiac Metabolism”. [Copyright &y& Elsevier]

Alternative cleavage and polyadenylation (APA) generates diverse mRNA isoforms. We developed 3? region extraction and deep sequencing (3?READS) to address mispriming issues that commonly plague poly(A) site (pA) identification, and we used the method to comprehensively map pAs in the mouse genome. Thorough annotation of gene 3? ends revealed over 5,000 previously overlooked pAs (?8% of total) flanked by A-rich sequences, underscoring the necessity of using an accurate tool for pA mapping. About 79% of mRNA genes and 66% of long noncoding RNA genes undergo APA, but these two gene types have distinct usage patterns for pAs in introns and upstream exons. Quantitative analysis of APA isoforms by 3?READS indicated that promoter-distal pAs, regardless of intron or exon locations, become more abundant during embryonic development and cell differentiation and that upregulated isoforms have stronger pAs, suggesting global modulation of the 3? end-processing activity in development and differentiation. [ABSTRACT FROM AUTHOR]

Adenylyl cyclase type 5 knockout mice ( AC5 KO) live longer and are stress resistant, similar to calorie restriction ( CR). AC5 KO mice eat more, but actually weigh less and accumulate less fat compared with WT mice. CR applied to AC5 KO results in rapid decrease in body weight, metabolic deterioration, and death. These data suggest that despite restricted food intake in CR, but augmented food intake in AC5 KO, the two models affect longevity and metabolism similarly. To determine shared molecular mechanisms, m RNA expression was examined genome-wide for brain, heart, skeletal muscle, and liver. Significantly more genes were regulated commonly rather than oppositely in all the tissues in both models, indicating commonality between AC5 KO and CR. Gene ontology analysis identified many significantly regulated, tissue-specific pathways shared by the two models, including sensory perception in heart and brain, muscle function in skeletal muscle, and lipid metabolism in liver. Moreover, when comparing gene expression changes in the heart under stress, the glutathione regulatory pathway was consistently upregulated in the longevity models but downregulated with stress. In addition, AC5 and CR shared changes in genes and proteins involved in the regulation of longevity and stress resistance, including Sirt1, Apo D, and olfactory receptors in both young- and intermediate-age mice. Thus, the similarly regulated genes and pathways in AC5 KO and CR mice, particularly related to the metabolic phenotype, suggest a unified theory for longevity and stress resistance. [ABSTRACT FROM AUTHOR]

The purpose of this study was to develop a new formulation of intravenous lipid microspheres for bufadienolides (bufadienolides-loaded lipid microspheres, BU-LM) and investigate the stability of BU-LM after dilution and long-term storage. Photon correlation spectroscopy, electrophoretic light scattering, and light microscopy were used to evaluate the physical characteristics of BU-LM. The chemical properties of bufadienolides were measured by high-performance liquid chromatography. The optimum BU-LM, consisting of 10% MCT, 3.0% Lipoid S75, 0.4% F68, 0.05% sodium oleate and 2.5% glycerol, was prepared by emulsification with a high-speed shear mixing at 10 000 rpm for 12 min and a high-pressure homogenization at 800 bar for 8 cycles at 30°C. The BU-LM at pH 6.48, with the particle size distribution of 175.2 ± 57.3 nm, ζ-potential of −32.10 mV, entrapment efficiency of 86.9%, 91.7%, 92.9%, and contents of 20.8%, 31.3%, 50.4% for B, C, R, respectively, was stable after sterilization in a 121°C rotating autoclave water steam bath for 10 min. Moreover, BU-LM had sufficient physicochemical stability after dilution with 5% glucose injection and long-term storage at 4 ± 2°C for 18 months. BU-LM has a great potential for clinical applications as a new drug delivery system for bufadienolides. Practical applications: Bufadienolides, mainly composed of bufalin (B), cinobufagin (C), and resibufogenin (R) have poor aqueous solubility and are chemically unstable. Currently the main marketable products of bufadienolides are solution injections, which cause serious venous irritation and in this formulation, the drug is not protected from degradation. Lipid microspheres, with the unique properties of a low cost, low toxicity, good storage stability, easy to scale up production, sustained release and targeting effect, incorporate the drugs into the interior oil phase and the oil-water interfacial film; thus a direct contact of the drug with body fluids and tissues is prevented, which increases the stability of the drugs and minimizes the possible side effects. [ABSTRACT FROM AUTHOR]

PARN is one of several deadenylase enzymes present in mammalian cells, and as such the contribution it makes to the regulation of gene expression is unclear. To address this, we performed global mRNA expression and half-life analysis on mouse myoblasts depleted of PARN. PARN knockdown resulted in the stabilization of 40 mRNAs, including that encoding the mRNA decay factor ZFP36L2. Additional experiments demonstrated that PARN knockdown induced an increase in Zfp36l2 poly(A) tail length as well as increased translation. The elements responsible for PARN-dependent regulation lie within the 3' UTR of the mRNA. Surprisingly, changes in mRNA stability showed an inverse correlation with mRNA abundance; stabilized transcripts showed either no change or a decrease in mRNA abundance. Moreover, we found that stabilized mRNAs had reduced accumulation of pre-mRNA, consistent with lower transcription rates. This presents compelling evidence for the coupling of mRNA decay and transcription to buffer mRNA abundances. Although PARN knockdown altered decay of relatively few mRNAs, there was a much larger effect on global gene expression. Many of the mRNAs whose abundance was reduced by PARN knockdown encode factors required for cell migration and adhesion. The biological relevance of this observation was demonstrated by the fact that PARN KD cells migrate faster in wound-healing assays. Collectively, these data indicate that PARN modulates decay of a defined set of mRNAs in mammalian cells and implicate this deadenylase in coordinating control of genes required for cell movement. [ABSTRACT FROM AUTHOR]

Breast milk is the primary source of nutrition for newborns, and is rich in immunological components. MicroRNAs (miRNAs) are present in various body fluids and are selectively packaged inside the exosomes, a type of membrane vesicles, secreted by most cell types. These exosomal miRNAs could be actively delivered into recipient cells, and could regulate target gene expression and recipient cell function. Here, we analyzed the lactation-related miRNA expression profiles in porcine milk exosomes across the entire lactation period (newborn to 28 days after birth) by a deep sequencing. We found that immune- related miRNAs are present and enriched in breast milk exosomes (p<10- 16, Χ² test) and are generally resistant to relatively harsh conditions. Notably, these exosomal miRNAs are present in higher numbers in the colostrums than in ature milk. It was higher in the serum of colostrum-only fed piglets compared with the mature milk-only fed piglets. These immune- related miRNA-loaded exosomes in breast milk may be transferred into the infant body via the digestive tract. These observations are a prelude to in-depth investigations of the essential roles of breast milk in the development of the infant's immune system. [ABSTRACT FROM AUTHOR]

Transcription termination of RNA polymerase II between closely spaced genes is an important, though poorly understood mechanism. This is true, in particular, in the Drosophila genome, where approximately 52% of tandem genes are separated by less than 1 kb. We show that a set of Drosophila tandem genes have a negative correlation of gene expression and displays several molecular marks indicative of promoter pausing polymerase. We find that an intergenic spacing of 168 bp is sufficient for efficient transcription termination between the polo-snap tandem gene pair, by a mechanism that is independent of Pcf11 and Xrn2. In contrast, analysis of a tandem gene pair containing a longer intergenic region reveals that termination occurs farther downstream of the poly(A) signal and is, in this case, dependent on Pcf11 and Xrn2. For polo-snap, displacement of poised polymerase from the snap promoter by depletion of the initiation factor TFIIB results in an increase of polo transcriptional read-through. This suggests that poised polymerase is necessary for transcription termination. Interestingly, we observe that polo forms a TFIIB-dependent gene loop between its promoter and terminator regions. Furthermore, in a plasmid containing the polo-snap locus, deletion of the polo promoter causes an increase in snap expression, as does deletion of polo poly(A) signals. Taken together, our results indicate that polo forms a gene loop and polo transcription termination occurs by an Xrn2 and Pcf11 independent mechanism that requires TFIIB. [ABSTRACT FROM AUTHOR]

Summary: Mediator complex is an integrative hub for transcriptional regulation. Here we show that Mediator regulates alternative mRNA processing via its MED23 subunit. Combining tandem affinity purification and mass spectrometry, we identified a number of mRNA processing factors that bind to a soluble recombinant Mediator subunit, MED23, but not to several other Mediator components. One of these factors, hnRNP L, specifically interacts with MED23 in vitro and in vivo. Consistently, Mediator partially colocalizes with hnRNP L and the splicing machinery in the cell. Functionally, MED23 regulates a subset of hnRNP L-targeted alternative splicing (AS) and alternative cleavage and polyadenylation (APA) events, as shown by minigene reporters and exon array analysis. ChIP-seq analysis revealed that MED23 can regulate hnRNP L occupancy at their coregulated genes. Taken together, these results demonstrate a crosstalk between Mediator and the splicing machinery, providing a molecular basis for coupling mRNA processing to transcription. [Copyright &y& Elsevier]

Summary: p53 is a frequent target for mutation in human tumors, and mutant p53 proteins can actively contribute to tumorigenesis. We employed a three-dimensional culture model in which nonmalignant breast epithelial cells form spheroids reminiscent of acinar structures found in vivo, whereas breast cancer cells display highly disorganized morphology. We found that mutant p53 depletion is sufficient to phenotypically revert breast cancer cells to a more acinar-like morphology. Genome-wide expression analysis identified the mevalonate pathway as significantly upregulated by mutant p53. Statins and sterol biosynthesis intermediates reveal that this pathway is both necessary and sufficient for the phenotypic effects of mutant p53 on breast tissue architecture. Mutant p53 associates with sterol gene promoters at least partly via SREBP transcription factors. Finally, p53 mutation correlates with highly expressed sterol biosynthesis genes in human breast tumors. These findings implicate the mevalonate pathway as a therapeutic target for tumors bearing mutations in p53. [Copyright &y& Elsevier]

Summary: BIK protein is an initiator of mitochondrial apoptosis, and BIK expression is induced by proapoptotic signals, including DNA damage. Here, we demonstrate that 3′ end processing and expression of BIK mRNA are controlled by the nuclear PI4,5P2-regulated poly(A) polymerase Star-PAP downstream of DNA damage. Nuclear PKCδ is a key mediator of apoptosis, and DNA damage stimulates PKCδ association with the Star-PAP complex where PKCδ is required for Star-PAP-dependent BIK expression. PKCδ binds the PI4,5P2-generating enzyme PIPKIα, which is essential for PKCδ interaction with the Star-PAP complex, and PKCδ activity is directly stimulated by PI4,5P2. Features in the BIK 3′ UTR uniquely define Star-PAP specificity and may block canonical PAP activity toward BIK mRNA. This reveals a nuclear phosphoinositide signaling nexus where PIPKIα, PI4,5P2, and PKCδ regulate Star-PAP control of BIK expression and induction of apoptosis. This pathway is distinct from the Star-PAP-mediated oxidative stress pathway indicating signal-specific regulation of mRNA 3′ end processing. [ABSTRACT FROM AUTHOR]

Well-ordered large-area two-dimensionally (2D) periodic and individually addressable metal–ferroelectric–metal (M/F/M) nanocapacitor arrays were fabricated by focused ion beam (FIB) milling with synchronal and serial mode. Scanning electron microscopy (SEM) results indicated that the synchronal mode can fabricate more uniform arrays than serial mode, but the surface of arrays is damaged heavily. The various gray contrasts of domain images can be clearly observed in the M/F/M arrays, and its piezoresponse signal is larger than that of the F/M arrays. Piezoresponse force microscope (PFM) results also indicate that the ion damage can be reduced by top Pt electrode, but cannot be avoided. The displacement-voltage and P-E loops of the M/F/M nanocapacitor are indicative of strong ferroelectricity for the measurements. The remanent polarization is reduced by ion damage, but the coercive field is not increased comparing to the thin film. [ABSTRACT FROM AUTHOR]

Cardiac hypertrophy is enlargement of the heart in response to physiological or pathological stimuli, chiefly involving growth of myocytes in size rather than in number. Previous studies have shown that the expression pattern of a group of genes in hypertrophied heart induced by pressure overload resembles that at the embryonic stage of heart development, a phenomenon known as activation of the ''fetal gene program''. Here, using a genome-wide approach we systematically defined genes and pathways regulated in short- and long-term cardiac hypertrophy conditions using mice with transverse aortic constriction (TAC), and compared them with those regulated at different stages of embryonic and postnatal development. In addition, exon-level analysis revealed widespread mRNA isoform changes during cardiac hypertrophy resulting from alternative usage of terminal or internal exons, some of which are also developmentally regulated and may be attributable to decreased expression of Fox-1 protein in cardiac hypertrophy. Genes with functions in certain pathways, such as cell adhesion and cell morphology, are more likely to be regulated by alternative splicing. Moreover, we found 39UTRs of mRNAs were generally shortened through alternative cleavage and polyadenylation in hypertrophy, and microRNA target genes were generally de-repressed, suggesting coordinated mechanisms to increase mRNA stability and protein production during hypertrophy. Taken together, our results comprehensively delineated gene and mRNA isoform regulation events in cardiac hypertrophy and revealed their relations to those in development, and suggested that modulation of mRNA isoform expression plays an importance role in heart remodeling under pressure overload. [ABSTRACT FROM AUTHOR]

Genes containing multiple pre-mRNA cleavage and polyadenylation sites, or polyA sites, express mRNA isoforms with variable 3' untranslated regions (UTRs). By systematic analysis of human and mouse transcriptomes, we found that short 3'UTR isoforms are relatively more abundant when genes are highly expressed whereas long 3'UTR isoforms are relatively more abundant when genes are lowly expressed. Reporter assays indicated that polyA site choice can be modulated by transcriptional activity through the gene promoter. Using global and reporter-based nuclear run-on assays, we found that RNA polymerase II is more likely to pause at the polyA site of highly expressed genes than that of lowly expressed ones. Moreover, highly expressed genes tend to have a lower level of nucleosome but higher H3K4me3 and H3K36me3 levels at promoter-proximal polyA sites relative to distal ones. Taken together, our results indicate that polyA site usage is generally coupled to transcriptional activity, leading to regulation of alternative polyadenylation by transcription. [ABSTRACT FROM AUTHOR]

Abstract: The Id1 and Id3 genes play major roles during cardiac development, despite their expression being confined to non-myocardial layers (endocardium–endothelium–epicardium). We previously described that Id1Id3 double knockout (dKO) mouse embryos die at mid-gestation from multiple cardiac defects, but early lethality precluded the studies of the roles of Id in the postnatal heart. To elucidate postnatal roles of Id genes, we ablated the Id3 gene and conditionally ablated the Id1 gene in the endothelium to generate conditional KO (cKO) embryos. We observed cardiac phenotypes at birth and at 6months of age. Half of the Id cKO mice died at birth. Postnatal demise was associated with cardiac enlargement and defects in the ventricular septum, trabeculation and vasculature. Surviving Id cKO mice exhibited fibrotic vasculature, cardiac enlargement and decreased cardiac function. An abnormal vascular response was also observed in the healing of excisional skin wounds of Id cKO mice. Expression patterns of vascular, fibrotic and hypertrophic markers were altered in the Id cKO hearts, but addition of Insulin-Like Growth Factor binding protein-3 (IGFbp3) reversed gene expression profiles of vascular and fibrotic, but not hypertrophic markers. Thus, ablation of Id genes in the vasculature leads to distinct postnatal cardiac phenotypes. These findings provide important insights into the role/s of the endocardial network of the endothelial lineage in the development of cardiac disease, and highlight IGFbp3 as a potential link between Id and its vascular effectors. [ABSTRACT FROM AUTHOR]

The lattice gas automation (LGA) simulations for electrical transport properties have been performed on digital rock samples virtually saturated with oil and water to investigate the relationship between resistivity index and water (oil) saturation ( I-Sw) of reservoir rocks at micropore scale. The digital rocks were constructed by the pileup of matrix grains with a radius distribution obtained by laboratory measurements on reservoir rock samples. The LGA was then applied to simulate the flow of electrical current through the pores of these virtually saturated digital rocks to reveal the non-Archie relation of I-Sw. The results from LGA simulation indicate that the I-Sw relation is generally a non-linear function changing with the decrease of water saturation and porosity on a log-log scale. Archie's equation is an approximation in high water saturation range. Based on this study, we developed a new equation for non-Archie relation of I-Sw to improve the calculation of pore fluid saturation. The calculated results with this new equation have shown better fit to laboratory measurements. [ABSTRACT FROM AUTHOR]

Abstract: The aim of the present work is to develop high azithromycin loading powders for inhalation with optimal physical and aerodynamic properties, and to test their in vivo potential in rats. A five-level three-factorial central composite rotatable design was used for conducting the experiments. The optimized powder, which had a better flowability, an aerodynamic diameter of 3.82μm and an in vitro deposition of 51%, was obtained under selected spray-drying conditions. The content of the powder was high (59.2%), which met the requirement for high drug loading. The optimized powder was further examined in the in vivo study in rats. Lung microdialysis and blood microdialysis were simultaneously performed on each rat. The ratio of the AUCELF value between the intratracheal route and intravenous injection was 161.6, whereas the absolute bioavailability was only 43.5%, and the drug targeting index of the intratracheal route was 486.2. The results showed that azithromycin dry powder inhalers (DPI) could be effectively and efficiently delivered to a specific target site and achieve a high local concentration. In conclusion, AZI DPI offers an attractive alternative that is able to deliver high concentrations of antibiotic directly to the chosen target site while minimizing the systemic bioavailability and side effects. [Copyright &y& Elsevier]

Objectives: The aim of this study was to investigate whether dendritic cells (DCs) transfected with human secondary lymphoid-tissue chemokine (hSLC) and human interleukin-2 (hIL-2) genes are capable of improving DC's proliferation and to produce a marked antitumor effect in vitro combined with T-lymphocyte (TC). Methods: SLC gene primer was designed based on the corresponding gene sequence in GenBank. The Kpn I site was introduced into the upstream of the primer and Xho I site into the downstream. The SLC gene was amplified with the template of pET32a(+)-SLC by polymerase chain reaction. SLC was cloned into pBudCE4.1/IL-2 (TRAIL was cut from pBudCE4.1/TRAIL- IL-2 before) to construct recombinant plasmid pBudCE4.1/SLC-IL-2(PSI). DCs were transfected with pBudCE4.1/SLC-IL-2 by gene electric transfection. Protein expression was determined with Western blot and enzyme-linked immunosorbent assays. Cytotoxicity of TC and DC against the human bladder tumor cell were examined by chromium release assay. Flow cytometric analyses were performed to determine the apoptosis of tumor cells and the percentage of Treg. Results: A high level of expression of SLC and IL-2 was observed in DCs transfected with SLC and IL-2 genes. The mean production of IL-2 was 19.8 ± 2.5, 511.10 ± 52.36, and 541.3 ± 62.04 ng/106 cells/24 hours in the DC/vector, DC/IL-2, and DC/SLC-IL-2, respectively. The mean SLC production was 29.8 ± 4.43, 506.10 ± 42.36, and 567.34 ± 52.05 ngs/106cells/24 hours in the DC/ vector, DC/SLC, and DC/SLC-IL-2, respectively. Cytotoxicity to bladder cancer cells was increased. The mean cytotoxicity (the effector/target ratio, 40:1) of TC-DC/parental, TC-DC/IL-2, TC-DC/SLC, and TC-DC/SLC-IL-2(TDSI) to the human bladder cancer cells was 32.1 ± 5.5%, 63.5 ± 6.6%, 78.1 ± 9.63%, respectively. The apoptotsis rate of bladder cancer cells treated with TDSI was 18.6% by flow cytometry. Treg cells' percentage was very small in the DC medium. Conclusions: SLC and IL-2 were produced by autocrine in DCs transfected with SLC and IL-2 genes. DC/SLC-IL-2 can promote DC proliferation, while TC-DC/SLC-IL-2 and TC-DC/SLC could strongly enhance significant cytotoxicity against bladder cancer cell that was induced by the coculture of DCs (transfected with SLC and IL-2) and TC. [ABSTRACT FROM AUTHOR]

Abstract: The symplectic geometry method is introduced for exact bending solutions of moderately thick rectangular plates with two opposite edges simply supported. The basic equations for the plates are first transferred into Hamilton canonical equations. The whole state variables are then separated. Using the method of eigenfunction expansion in the symplectic geometry, typical examples for plates with selected boundary conditions are solved and exact bending solutions obtained. Since only the basic elasticity equations of the plates are used, this method eliminates the need to pre-determine the deformation function and is hence more reasonable than conventional methods. Numerical results were presented to demonstrate the validity and accuracy of this approach as compared to those reported in other literatures. [Copyright &y& Elsevier]

A single layer of CoFeB and a multilayer of CoFeB-MgO films are prepared by means of DC/RF magnetron sputter deposition. The excellent microwave properties and high electrical resistivity are simultaneously achieved in the discontinuous multilayer structure of [Co44Fe44B12(0.7 nm)/MgO(0.4 nm)]40 film. This film has a high permeability (m'') (larger than 100 below 2.1 GHz), a high magnetic loss (m'') (larger than 100 in a range from 1.5 to 3.3 GHz), a resistivity of 3.3 x 103mO*cm, a saturation magnetization of 1.2 T, and an in-plane uniaxial anisotropy field of 5.5 kA/m. The microstructure and the surface topography of the film are also analysed. The relatively large surface roughness for the discontinuous film is responsible for the wide frequency band of magnetic loss. The magnetic loss and the high resistivity indicate that the discontinuous CoFeB-MgO multilayer film has potential applications in microwave absorbers and electromagnetic interference (EMI) shielding materials in a GHz frequency range. [ABSTRACT FROM AUTHOR]

Urban heatwaves increase residential health risks. Identifying urban residential sensitivity to heatwave risks is an important prerequisite for mitigating the risks through urban planning practices. This research proposes a new paradigm for urban residential sensitivity to heatwave risks based on social media Big Data, and describes empirical research in five megacities in China, namely, Beijing, Nanjing, Wuhan, Xi'an and Guangzhou, which explores the application of this paradigm to real-world environments. Specifically, a method to identify urban residential sensitive to heatwave risks was developed by using natural language processing (NLP) technology. Then, based on remote sensing images and Weibo data, from the perspective of the relationship between people (group perception) and the ground (meteorological temperature), the relationship between high temperature and crowd sensitivity in geographic space was studied. Spatial patterns of the residential sensitivity to heatwaves over the study area were characterized at fine scales, using the information extracted from remote sensing information, spatial analysis, and time series analysis. The results showed that the observed residential sensitivity to urban heatwave events (HWEs), extracted from Weibo data (Chinese Twitter), best matched the temporal trends of HWEs in geographic space. At the same time, the spatial distribution of observed residential sensitivity to HWEs in the cities had similar characteristics, with low sensitivity in the urban center but higher sensitivity in the countryside. This research illustrates the benefits of applying multi-source Big Data and intelligent analysis technologies to the understand of impacts of heatwave events on residential life, and provide decision-making data for urban planning and management. [ABSTRACT FROM AUTHOR]

Duck hepatitis A virus 1 (DHAV-1) can cause high morbidity and fatal acute infectious hepatitis in ducklings, which seriously endangers animal husbandry. Viroporin is a small molecular weight hydrophobic transmembrane protein encoded by the virus, that has been suggested to induce autophagy in host cells by increasing the membrane permeability through disturbing the ion balance. In this study, we aimed to investigate whether the DHAV-1 2B protein can induce autophagy in DEF cells with a viroporin-like function. Bioinformatics analysis has indicated that the 2B protein is characterized by a viroporin domain, which is consistent with the type IA viroporin transmembrane protein. We experimentally confirmed that the 2B protein disturbed the Ca2+ balance of infected cells by elevating the intracellular Ca2+ concentration. Eukaryotic expression of the 2B protein upregulates the expression of microtubule-associated protein 1 light chain 3 II (LC3-II) and the number of autophagosomes in the cell. Interestingly, the Western Blot (WB) results showed that 2B protein expression induced less protein degradation of the autophagic substrate sequestosome 1 (SQSTM1/p62) than the positive control, while microscopy observations showed that the autophagosomes did not colocalize with the lysosomes. In summary, 2B protein expression induced autophagy in host cells, but the autophagic flow was incomplete. The results of this experiment are expected to provide reference scientific data for elucidating the infective and pathogenic mechanism of DHAV-1. [ABSTRACT FROM AUTHOR]

The feasibility of using respirable bacteriophage (phage) powder to treat lung infections has been demonstrated in animal models and clinical studies. This work investigated the influence of formulation compositions and excipient concentrations on the aerosol performance and storage stability of phage powder. An anti-Acinetobacter baumannii phage vB_AbaM-IME-AB406 was incorporated into dry powders consisting of trehalose, mannitol and L-leucine for the first time. The phage stability upon the spray-drying process, room temperature storage and powder dispersion under different humidity conditions were assessed. In general, powders prepared with higher mannitol content (40% of the total solids) showed a lower degree of particle merging and no sense of stickiness during sample handling. These formulations also provided better storage stability of phage with no further titer loss after 1 month and <1 log titer loss in 6 months at high excipient concentration. Mannitol improved the dispersibility of phage powders, but the in vitro lung dose dropped sharply after exposure to high-humidity condition (65% RH) for formulations with 20% mannitol. While previously collected knowledge on phage powder preparation could be largely extended to formulate A. baumannii phage into inhalable dry powders, the environmental humidity may have great impacts on the stability and dispersion of phage; therefore, specific attention is required when optimizing phage powder formulations for global distribution. [ABSTRACT FROM AUTHOR]

The design and development of multifunctional nanoparticles have attracted great interest in biomedical research. This study aims to prepare pH-responsive melanin-like nanoparticles for T1-weighted magnetic resonance imaging (MRI) and photothermal therapy. The new multifunctional nanoparticles (amino-Fe-PDANPs) are synthesized by copolymerization of dopamine and its derivative amino-N-[2-(diethylamino) ethyl]-3,4-dihydroxy-benzenepropanamide (N-Dopa) at room temperature. The size of nanoparticles can be controlled by NaOH concentration. The incorporation of N-Dopa is characterized by NMR and FT-IR. From transmission electron microscopy (TEM), the nanoparticles exhibit excellent dispersion stability in water and are spherical in shape. The MRI measurement has demonstrated that amino-Fe-PDANPs have a significant signal enhancement in responding to the acidic solution. Confirmed by the photothermal study, the nanoparticles exhibit a high photothermal conversion efficiency. The melanin-like multifunctional nanoparticles integrate both diagnosis and therapeutic functionalities, indicating the potential for theranostic application. [ABSTRACT FROM AUTHOR]

Myocardial infarction (MI) is often followed by heart failure (HF), but the mechanisms precipitating the transition to HF remain largely unknown. A genomic profile was performed in a monkey model of MI, from the myocardium adjacent to chronic (2-month) MI followed by 3 weeks of pacing to develop HF. The transcript of the gene encoding the cell cycle-related kinase (CCRK) was down-regulated by 50% in HF heart compared with control (p < 0.05), which was confirmed by quantitative PCR. The CCRK sequence cloned from a heart library showed a con- servation of the N-terminal kinase domain when compared with the "generic" isoform cloned previously but a different C-terminal half due to alternative splicing with frameshift. The homology of the cardiac sequence was 100% between mice and humans. Expression of the corresponding protein, measured upon generation of a monoclonal antibody, was limited to heart, liver, and kidney. Upon overexpression in cardiac myocytes, both isoforms promote cell growth and reduce apoptosis by chelerythrine (p < 0.05 versus control). Using a yeast two-hybrid screening, we found an interaction of the generic but not the cardiac CCRK with cyclin H and casein kinase 2. In addition, only the generic CCRK phosphorylates the cyclin-dependent kinase 2, which was accompanied by a doubling of myocytes in the S and G2 phases of the cell cycle (p < 0.05 versus control). Therefore, the heart expresses a splice variant of CCRK, which promotes cardiac cell growth and survival; differs from the generic isoform in terms of protein-protein interactions, substrate specificity and regulation of the cell cycle; and is down-regulated significantly in HF. [ABSTRACT FROM AUTHOR]

Abstract: Based on the promising opioid pharmacological profile of the peptide, Tyr-Pro-Trp-Gly-NH2 (Tyr-W-MIF), Zadina et al. [Zadina, J.E., Hackler, L., Ge, L.-J., Kastin, A.J., 1997. A potent and selective endogenous agonist for the μ-opiate receptor. Nature 386, 499–5502] synthesized and screened other Gly4-substituted peptides, culminating in the synthesis of Tyr-Pro-Trp-Phe-NH2 (endomorphin-1), which displayed high affinity and selectivity for the μ-opioid receptor. The amidated peptide was then isolated from bovine brain frontal cortex, as was a related peptide, Tyr-Pro-Phe-Phe-NH2 (endomorphin-2), that displayed similar high affinity and selectivity for the μ-opioid receptor. The biosynthesis of the endomorphins in the brain remains obscure, since the putative precursor proteins for the peptides have not been identified. With the completion of the human genome sequencing project, we hypothesized that we should uncover the biological precursors of the peptides using a bioinformatic approach to search the current human proteome for proteins that contained the endomorphin peptide sequences followed by Gly-Lys/Arg, the consensus sequence for peptide α-amidation and precursor cleavage. Twelve proteins were identified that contained the endomorphin-1 Tyr-Pro-Trp-Phe sequence, however none contained the Tyr-Pro-Trp-Phe-Gly sequence necessary for α-amidation. Twenty-two distinct proteins contained the endomorphin-2 tetrapeptide sequence, and two of those contained the sequence, Tyr-Pro-Phe-Phe-Gly, however, none contained the requisite peptide-Gly-Lys/Arg sequence. Western blot analysis using an endomorphin-2 antibody detected 4 prominent proteins in mouse brain, necessitating reinterpretation of previous immunocytolocalization studies in the brain. Screening of the current human proteome yielded no evidence for endomorphin precursor proteins based on accepted biochemical criteria. [Copyright &y& Elsevier]

Abstract: Human CstF-77 is one of the three subunits of cleavage stimulation factor (CstF) that is essential for mRNA polyadenylation. Its Drosophila homologue, suppressor of forked [su(f)], contains an intronic poly(A) site, which can lead to a short transcript without a stop codon. By both bioinformatic searches and validation with molecular biology experiments, we found that human and mouse CstF-77 genes also contain an intronic poly(A) site, which can be utilized to produce short CstF-77 transcripts lacking sequences encoding domains that are involved in many of the CstF-77 functions. The genomic sequence surrounding the poly(A) site is highly conserved among all vertebrates, but is not present in non-vertebrate species. Using public Serial Analysis of Gene Expression (SAGE) data, we found that the intronic poly(A) site is utilized in a wide range of tissues. This finding indicates that vertebrates may employ a similar alternative polyadenylation mechanism to modulate CstF-77, highlighting the importance of the regulation of CstF-77 in various species. [Copyright &y& Elsevier]

Distribution and speciation of heavy metals of agricultural soils (85 surface soil samples and 4 soil profiles) in Dongguan were investigated, while total Cr, Cu, Ni, Pb, Zn (abbreviated as Cr, Cu, Ni, Pb, Zn) and available Cu, Zn (Av-Cu, Av-Zn) were analyzed by a flame absorption spectrophotometer (AAS), and total Cd (Cd) was analyzed using graphite furnace AAS. The content of Cd, Cu and Ni was partially much more than the second grade of GB15618-1995 even though the mean contents of all heavy metal were less than the threshold value of the second grade and only the mean content of Pb was more than the value of national background. Results of descriptive statistic showed that the mean content of heavy metals should depend on land utilization and spatial location at some extent. The heavy metal contents were higher in the Southwest and Northwest than in the Central. In addition, the mean contents of Zn and Pb in Dongguan paddy soils were significantly higher than those of Pearl River Delta (PRD) and Taihu Lake region (TLR). Correlation analyses indicated that there existed significant correlation between Cr and Ni in orchard soils, and among Zn, Cd and Cu, between Av-Cu and Cu, between Av-Zn and Cr, Ni, pH value in vegetable soils, and a weak relationship between Cd, Cu and Pb, between Av-Zn and Zn. Principal component analyses (PCA) showed that the order of importance should be Zn> Pb> Cr> Ni> Cu. [ABSTRACT FROM AUTHOR]

There are limited studies attempting to correlate the expression changes in oral squamous cell carcinoma with clinically relevant variables. We determined the gene expression profile of 16 tumor and 4 normal tissues from 16 patients by means of Affymetrix Hu133A GeneChips. The hybridized RNA was isolated from cells obtained with laser capture microdissection, then was amplified and labeled using T7 polymerase-based in vitro transcription. The expression of 53 genes was found to differ significantly (33 upregulated, 20 downregulated) in normal versus tumor tissues under two independent statistical methods. The expression changes in four selected genes (LGALS1, MMP1, LAGY, and KRT4) were confirmed with reverse transcriptase polymerase chain reaction. Two-dimensional hierarchical clustering of the 53 genes resulted in the samples clustering according to the extent of tumor infiltration: normal epithelial tissue, tumors less than or equal to 4 cm in dimension, and tumors more than 4 cm in dimension (P=0.0014). The same pattern of clustering was also observed for the 20 downregulated genes. We did not observe any associations with lymph node metastasis (P=0.097). [Copyright &y& Elsevier]