A novel BRET based genetic coded biosensor for apoptosis detection at deep tissue level in live animal.

ANNEXIN V belongs to a family of phospholipid binding proteins which is able to bind to negatively charged phospholipids such as phosphatidylserine (PS) in the presence of a high affinity Ca2+ ion. When apoptosis occurs, even at early stage, PS will be exposed to the outside of the cell surface from the cytoplasm side of membrane leaflets., Therefore ANNEXIN V has been suggested as a bio-marker for imaging early apoptotic events of various cell death including those in disease conditions. However, most ANNEXIN V-based apoptotic detecting techniques were in vitro approaches. Here, we presented a new BRET (Bioluminescence Resonance Energy Transfer) based genetic coded biosensor by fusing ANNEXIN V and a BRET version of NanoLuc (teLuc) for both in vitro and in vivo apoptosis detection. The BRET feature of this new sensor makes it convenient to be applied to both conventional fluorescent-based in vitro apoptosis detection and bioluminescence-based animal live imaging for in vivo study. Because of its robust bioluminescence signal, it is possible to perform the evaluation of the disease-induced apoptotic damage and recovery process directly at deep tissue level in live animal.

Lattice Dynamics and Thermal Conductivity in Cu2Zn1- xCo xSnSe4.

The quaternary compound Cu2ZnSnSe4 (CZTSe), as a typical candidate for both solar cells and thermoelectrics, is of great interest for energy harvesting applications. Materials with a high thermoelectric efficiency have a relatively low thermal conductivity, which is closely related to their chemical bonding and lattice dynamics. Therefore, it is essential to investigate the lattice dynamics of materials to further improve their thermoelectric efficiency. Here we report a lattice dynamic study in a cobalt-substituted CZTSe system using temperature-dependent X-ray absorption fine structure spectroscopy (TXAFS). The lattice contribution to the thermal conductivity is dominant, and its reduction is mainly ascribed to the increment of point defects after cobalt substitution. Furthermore, a lattice dynamic study shows that the Einstein temperature of atomic pairs is reduced after cobalt substitution, revealing that increasing local structure disorder and weakened bonding for each of the atomic pairs are achieved, which gives us a new perspective for understanding the behavior of lattice thermal conductivity.

Rhythmic expressed clock regulates the transcription of proliferating cellular nuclear antigen in teleost retina.

Teleost fish continues to grow their eyes throughout life with the body size. In Astatotilapia burtoni, the fish retina increases by adding new retinal cells at the ciliary marginal zone (CMZ) and in the outer nuclear layer (ONL). Cell proliferation at both sites exhibits a daily rhythm in number of dividing cells. To understand how this diurnal rhythm of new cell production is controlled in retinal progenitor cells, we studied the transcription pattern of clock genes in retina, including clock1a, clock1b, bmal1a (brain and muscle ARNT-Like), and per1b (period1b). We found that these genes have a strong diurnal rhythmic transcription during light-dark cycles but not in constant darkness. An oscillation in pcna transcription was also observed during light-dark cycles, but again not in constant darkness. Our results also indicate an association between Clock proteins and the upstream region of pcna (proliferating cellular nuclear antigen) gene. A luciferase reporter assay conducted in an inducible clock knockdown cell line further demonstrated that the mutation on predicted E-Boxes in pcna promoter region significantly attenuated the transcriptional activation induced by Clock protein. These results suggested that the diurnal rhythmic expression of clock genes in A. burtoni retina could be light dependent and might contribute to the daily regulation of the proliferation of the retina progenitors through key components of cell cycle machinery, for instance, pcna.

Nanoscale heterogeneity in thermoelectrics: the occurrence of phase separation in Fe-doped Ca3Co4O9.

The misfit layered cobaltate thermoelectrics are good candidates for high temperature thermoelectric applications. Ca3Co4O9 is a typical compound of this family, which consists of rock salt Ca2CoO3 slabs alternating with hexagonal CoO2 slabs with a large lattice mismatch along the b axis. Each slab is 0.3-0.5 nm thick and shows an inherent structural heterogeneity at the nanoscale. The latter is a key parameter that affects the electrical transport and the heat flow in these misfit structured thermoelectrics. To clarify the physical origin of the thermoelectric performance of iron doped Ca3Co4O9 we combined X-ray near-edge absorption spectroscopy (XANES) and quantum modeling using density functional theory. In contrast to single-site doping, the iron doping first occurs at the Co1 site of the rock salt slab at low doping while at higher doping it prefers the Ca1 site of the rock salt slab. Doping at the Ca1 site modifies the electronic structure tuning the nanoscale structural heterogeneity. This mechanism may open a new route to optimizing the thermoelectric performance of misfit layered thermoelectrics.

Enhanced thermoelectric properties of Ga-doped In2O3 ceramics via synergistic band gap engineering and phonon suppression.

Ga doped In2O3-based thermoelectric materials were prepared by spark plasma sintering (SPS) using sintered powders in the low temperature solid phase. The solubility of Ga in In2O3 is about 10 at%, much larger than other elements such as Ge, Ce, etc. The larger solubility of Ga allows us to optimize the thermal and electrical transport properties of Ga doped In2O3 in a wider window. While tuning the concentration of dopants, the thermoelectric performance of Ga doped In2O3 was enhanced through a synergistic approach combining band-gap engineering and phonon suppression. The power factor increases from ∼0.5 × 10(-4) to ∼9.6 × 10(-4) W mK(-2) at 700 °C while thermal conductivity reduces from ∼4 to ∼2 W mK(-1) at 700 °C in In1.9Ga0.1O3. The maximum ZT of 0.37, increased by a factor of 4 from the pristine In2O3, is achieved in In1.9Ga0.1O3 at 700 °C.

Effect of psyllium husk powder on the gelation behavior, microstructure, and intermolecular interactions in myofibrillar protein gels from Andrias davidianus.

The interaction between proteins and soluble dietary fibers plays a vital role in the development of animal-derived foods. Herein, the effects of different contents (0-3.0%) of round-bracted psyllium husk powder (PHP) on the gelation behavior, microstructure, and intermolecular interactions of Andrias davidianus myofibrillar protein (MP) were investigated. Rheological and chemical forces suggested that PHP (1.5%-2.0%) enhanced the functional properties of MP at low ionic strength, thereby increasing the viscoelasticity of mixed gels. SDS-PAGE revealed that PHP reinforced the cross-linking and aggregation of protein molecules. Circular dichroism spectroscopy, low-field nuclear magnetic resonance, and scanning electron microscopy demonstrated that PHP induced the transformation of α-helix (decreased by 14.85%) to an ordered ß-sheet structure (increased by 81.58%), which was more favorable for the formation of dense network structure and improved (10.53%) the water retention of MP gels. This study provided new insights for PHP to effectively meliorate the heat-induced gelling properties of MP.

Microstructure of microemulsion modified with ionic liquids in microemulsion electrokinetic chromatography and analysis of seven corticosteroids.

In this work, the influences of ionic liquid (IL) as a modifier on microemulsion microstructure and separation performance in MEEKC were investigated. Experimental results showed that synergetic effect between IL 1-butyl-3-methylimidazolium tetrafluoro-borate (BmimBF4 ) and surfactant SDS gave a decreased CMC. With increment of IL in microemulsion, negative ζ potential of the microdroplets reduced gradually. The influence of IL on the dimensions of microdroplet was complicated. At BmimBF4 less than 8 mM, IL made microemulsion droplet smaller in size. While at BmimBF4 more than 10 mM, the size increased and reached to a maximum value at 12 mM, where the microdroplets were larger than that without IL. After that, the micreodroplet size decreased again. Relative fluorescence intensity of the first vibration band of pyrene to the third one (I1 /I3 ) enhanced as IL was added to microemulsion, which indicated that this addition increased environmental polarity in the inner core of microdroplets. Prednisone, hydrocortisone, prednisolone, hydrocortisone acetate, cortisone acetate, prednisolone acetate, and triamcinolone acetonide were analyzed with MEEKC modified with IL to evaluate the separation performance. Cortisone acetate and prednisolone acetate could not be separated at all in typical microemulsion. The seven analytes could be separated by the addition of 10 mM BmimBF4 into the microemulsion system. The method has been used for analysis of corticosteroids in cosmetic samples with simple extraction; the recoveries for seven analytes were between 86 and 114%. This method provides accuracy, reproducibility, pretreatment simplicity, and could be applied to the quality control of cosmetics.

Restoration of muscle fibers and satellite cells after isogenic MSC transplantation with microdystrophin gene delivery.

Duchenne muscular dystrophy is the most prevalent inheritable muscle disease. Transplantation of autologous stem cells with gene direction is an ideal therapeutic approach for the disease. The current study aimed to investigate the restoration of myofibers in mdx mice after mdx bone marrow-derived mesenchymal stem cell (mMSC) transplantation with human microdystrophin delivery. Possible mechanisms of action were also studied. In our research, mMSCs were successfully transduced by retrovirus carrying a functional human microdystrophin gene. Transplantation of transduced mMSCs enabled persistent dystrophin restoration in the skeletal muscle of mdx mice up to the 12th week after transplantation. Simultaneous coexpression of human microdystrophin and desmin showed that implanted mMSCs are capable of long-term survival as muscle satellite cells.

Potential correlation between volatiles and microbiome of Xiang xi sausages from four different regions.

Volatiles and microbiome from different Xiang xi sausages (HBD, HBN, HHY and HDK) were performed by Chromatography-Ion Mobility Spectrometry and Illumina MiSeq platform (16 S rRNA and ITS sequencing). Results indicated that ethanol, ethyl acetate, ethyl-propanoate, pinene, limonene, cineole, etc. were the key (main) volatiles; whereas staphylococcus, lactobacillus, solanum-torvum, brochothrix and debaryomyces, candida were the dominant bacteria and fungi, respectively. The variation and relationship between microbiome and volatiles were disclosed by multivariate and spearman's correlation analysis. Volatiles of HDK is similar to HBN and HBD compared to HHY, while microbiome community of HHY is similar to HBN and HDK compared to HBD. Solanum-torvum, pseudomonas (bacteria) and debaryomyces (fungi) contributed positively to the development of major volatiles in Xiangxi sausages. Meanwhile, 'carbohydrate metabolism', 'amino acid metabolism' (microbia) and glycolysis/gluconeogenesis (volatiles) were commented in KEGG database. This study provides a deep insight into the relation between volatiles and microbiome (bacteria and fungi) of Xiangxi sausages.

Partial epilepsy with antecedent febrile seizures and seizure aggravation by antiepileptic drugs: associated with loss of function of Na(v) 1.1.

PURPOSE: Generalized epilepsy with febrile seizures plus (GEFS+) and severe myoclonic epilepsy in infancy (SMEI) are associated with sodium channel α-subunit type-1 gene (SCN1A) mutations. Febrile seizures and partial seizures occur in both GEFS+ and SMEI; sporadic onset and seizure aggravation by antiepileptic drugs (AEDs) are features of SMEI. We thus searched gene mutations in isolated cases of partial epilepsy with antecedent FS (PEFS+) that showed seizure aggravations by AEDs. METHODS: Genomic DNA from four patients was screened for mutations in SCN1A, SCN2A, SCN1B, and GABRG2 using denaturing high-performance liquid chromatography (dHPLC) and sequencing. Whole-cell patch clamp analysis was used to characterize biophysical properties of two newly defined mutants of Na(v) 1.1 in tsA201 cells. RESULTS: Two heterozygous de novo mutations of SCN1A (R946H and F1765L) were detected, which were proven to cause loss of function of Na(v) 1.1. When the functional defects of mutants reported previously are compared, it is found that all mutants from PEFS+ have features of loss of function, whereas GEFS+ shows mild dysfunction excluding loss of function, coincident with mild clinical manifestations. PEFS+ is similar to SMEI clinically with possible AED-induced seizure aggravation and biophysiologically with features of loss of function, and different from SMEI by missense mutation without changes in hydrophobicity or polarity of the residues. CONCLUSIONS: Isolated milder PEFS+ may associate with SCN1A mutations and loss of function of Na(v) 1.1, which may be the basis of seizure aggravation by sodium channel-blocking AEDs. This study characterized phenotypes biologically, which may be helpful in understanding the pathophysiologic basis, and further in management of the disease.

Metal impurities dominate the sorption of a commercially available carbon nanotube for Pb(II) from water.

Numerous studies suggested carbon nanotubes (CNTs) as a type of promising sorbent for heavy metals from water and explained the sorption mechanism mainly by oxygen-containing functional groups on CNT surfaces but neglected the potential role of metal catalyst residues in CNTs. This is a first study showing that metal impurities could dominate the sorption of one type of commercially available CNTs (P-CNTs) for Pb(II) from water, which will help to understand and guide environmental applications of CNTs as a sorbent. Sorption capacity of P-CNTs (27.3 mg g(-1)) for Pb(II) was much higher than that of the water-washed P-CNTs (W-CNTs, 4.7 mg g(-1)). SEM-EDS and ICP-MS analyses showed that both P-CNTs and W-CNTs contained metal impurities (mainly Co and Mo) which released into the solutions during the sorption, especially P-CNTs. XAFS examination and precipitation experiments demonstrated that PbMoO(4) formation between Pb(II) and CNT-released MoO(4)(2-) and subsequent precipitation in the sorptive solutions was the dominant mechanism for the apparent sorption of Pb(II) by P-CNTs.

Bioremediation of Cr(VI) and immobilization as Cr(III) by Ochrobactrum anthropi.

Bioremediation of Cr(VI) through reduction relies on the notion that the produced Cr(III) may be precipitated or efficiently immobilized. However, recent reports suggest that soluble organo-Cr(III) complexes are present in various chromate-reducing bacterial systems. This work was designed to explore the factors that affect the immobilization of Cr(III) in the Ochrobactrum anthropi system. X-ray absorption fine structure analysis on the cell debris clearly verified that coordination of Cr(III) occurs on the surfaces via the chelating coordination with carboxyl- and amido-functional groups. However, competitive coordination experiments of Cr(III) revealed that the small molecules such as amino acids and their derivatives or multicarboxyl compounds hold stronger coordination ability with Cr(III) than with cell debris. We speculate that it is the preferential coordination of Cr(III) to the soluble organic molecules in the bacterial culture medium that inhibits effective immobilization of Cr(III) on the cells. On the basis of this understanding, a strategy with two-step control of the medium was proposed, and this achieved successful immobilization of Cr(VI) as Cr(III) by O. anthropi and Planococcus citreus in 5-50 L pilot-scale experiments.

Fractional-order modeling of lithium-ion batteries using additive noise assisted modeling and correlative information criterion.

In this paper, the fractional-order modeling of multiple groups of lithium-ion batteries with different states is discussed referring to electrochemical impedance spectroscopy (EIS) analysis and iterative learning identification method. The structure and parameters of the presented fractional-order equivalent circuit model (FO-ECM) are determined by EIS from electrochemical test. Based on the working condition test, a P-type iterative learning algorithm is applied to optimize certain selected model parameters in FO-ECM affected by polarization effect. What's more, considering the reliability of structure and adaptiveness of parameters in FO-ECM, a pre-tested nondestructive 1 / f noise is superimposed to the input current, and the correlative information criterion (CIC) is proposed by means of multiple correlations of each parameter and confidence eigen-voltages from weighted co-expression network analysis method. The tested batteries with different state of health (SOH) can be successfully simulated by FO-ECM with rarely need of calibration when excluding polarization effect. Particularly, the small value of CIC α indicates that the fractional-order α is constant over time for the purpose of SOH estimation. Meanwhile, the time-varying ohmic resistance R 0 in FO-ECM can be regarded as a wind vane of SOH due to the large value of CIC R 0 . The above analytically found parameter-state relations are highly consistent with the existing literature and empirical conclusions, which indicates the broad application prospects of this paper.

Speciation of zinc in secondary fly ashes of municipal solid waste at high temperatures.

The evaporation aerosols produced during the vitrification process of municipal solid waste incinerators (MSWI) fly ash represent a potential environmental risk owing to their high content of heavy metals. In this research, high-temperature heating processes were carried out on fly ashes collected from bag houses in a Chinese MSWI plant and the secondary fly ashes (SFA) were separately collected at three high temperatures (1273 K, 1423 K and 1523 K) below the melting range. Elemental analysis showed that high contents of both zinc and chlorine were present in these SFA samples and, according to the standard of the heavy metals industrial grade of ore, SFAs can be re-used as metallurgical raw materials or rich ore. Moreover, as shown by XAS analysis and for different high temperatures, zinc environments in the three SFA samples were characterized by the same local structure of the zinc chloride. As a consequence, a zinc recycling procedure can be easily designed based on the configuration information.

Comparative study of mesenchymal stem cells from C57BL/10 and mdx mice.

BACKGROUND: Human mesenchymal stem cells (MSCs) have been studied and applied extensively because of their ability to self-renew and differentiate into various cell types. Since most human diseases models are murine, mouse MSCs should have been studied in detail. The mdx mouse - a Duchenne muscular dystrophy model - was produced by introducing a point mutation in the dystrophin gene. To understand the role of dystrophin in MSCs, we compared MSCs from mdx and C57BL/10 mice, focusing particularly on the aspects of light and electron microscopic morphology, immunophenotyping, and differentiation potential. RESULTS: Our study showed that at passage 10, mdx-MSCs exhibited increased heterochromatin, larger vacuoles, and more lysosomes under electron microscopy compared to C57BL/10-MSCs. C57BL/10-MSCs formed a few myotubes, while mdx-MSCs did not at the same passages. By passage 21, mdx-MSCs but not C57BL/10-MSCs had gradually lost their proliferative ability. In addition, a significant difference in the expression of CD34, not Sca-1 and CD11b, was observed between the MSCs from the 2 mice. CONCLUSION: Our current study reveals that the MSCs from the 2 mice, namely, C57BL/10 and mdx, exhibit differences in proliferative and myogenic abilities. The results suggest that the changes in mouse MSC behavior may be influenced by lack of dystrophin protein in mdx mouse.

3D local structure around Zn in Kti11p as a representative Zn-(Cys)4 motif as obtained by MXAN.

Zinc is an important component of many proteins that play key roles in transcription, translation, and catalysis. Kti11p, DESR1, both belonging to a protein family characterized by a CSL zinc finger domain, and the co-catalytic zinc-protein PML containing a Zn(2+) binding domain called RING or C(3)HC(4) finger are all structurally determined by NMR although the zinc sites are silent to this spectroscopical method. The comparison of X-ray absorption near-edge spectroscopy (XANES) data for the three proteins demonstrates that fingerprints effect is a reliable method for a primary characterization of ligand species. Ab initio full MS calculations performed by MXAN are applied to obtain chemical and stereo structural information around the Zn ion in Kti11p. For the first time this high-spatial resolution technique confirms the formation of a stable Zn tetrahedral configuration with four sulfur ligands, and returns extremely accurate bond angle information between ligands.

[Construction of recombinant plasmid pVAX1-microdystrophin and preliminary study on the treatment to Duchenne muscular dystrophy].

OBJECTIVE: To construct the recombinant plasmid containing human microdystrophin cDNA, and study the microdystrophin expression in vivo and in vitro. METHODS: Microdystrophin cDNA was obtained from recombinant plasmid pBSK-MICRO digested with restrictive endonuclease Not I, the product was inserted into plasmid pVAX1, resulting in pAMICDYS. And then 3T3 cells were transfected with pAMICDYS. Forty-eight hours after transfection, the expression of the microdystrophin was detected by reverse transcription-polymerase chain reaction (RT-PCR) and immunocytochemistry. Finally, TA muscles of mdx mice were injected with the recombinant plasmid pAMICDYS through i.m. and the pathological change of TA was evaluated by histology, and the expression of microdystrophin in mdx TA was detected by immunohistochemical analysis. RESULTS: The recombinant plasmid containing human microdystrophin cDNA was constructed successfully. The recombinant plasmid was proved to be able to express microdystrophin protein both in vivo and in vitro. Moreover, treatment of the TA of mdx mice with the recombinant plasmid could decrease the number of centrally nucleated myofibers. CONCLUSION: Recombinant plasmid containing the microdystrophin gene was constructed successfully, and it could express microdystrophin protein both in vivo and in vitro. It provides basis for further study on microdystrophin as a target gene to treat Duchenne muscular dystrophy (DMD) by electrotransfer, i.v, arterial injection and combining with other exogenous gene to enhance microdystrophin expression.

A model based on feature objects aided strategy to evaluate the methane generation from food waste by anaerobic digestion.

A model based on feature objects (FOs) aided strategy was used to evaluate the methane generation from food waste by anaerobic digestion. The kinetics of feature objects was tested by the modified Gompertz model and the first-order kinetic model, and the first-order kinetic hydrolysis constants were used to estimate the reaction rate of homemade and actual food waste. The results showed that the methane yields of four feature objects were significantly different. The anaerobic digestion of homemade food waste and actual food waste had various methane yields and kinetic constants due to the different contents of FOs in food waste. Combining the kinetic equations with the multiple linear regression equation could well express the methane yield of food waste, as the R2 of food waste was more than 0.9. The predictive methane yields of the two actual food waste were 528.22 mL g-1 TS and 545.29 mL g-1 TS with the model, while the experimental values were 527.47 mL g-1 TS and 522.1 mL g-1 TS, respectively. The relative error between the experimental cumulative methane yields and the predicted cumulative methane yields were both less than 5%.

Chemical speciation of lead in secondary fly ash using X-ray absorption spectroscopy.

In this study, fly ash samples were collected from bag houses in a Chinese municipal solid waste incinerator (MSWI) and secondary fly ash (SFA) samples were collected from a high-temperature tubular electric furnace by thermal treatment of MSWI fly ash at 1050, 1100, 1150, 1200, and 1250 °C.We determined the speciation and atomic coordinates of lead in SFA using X-ray absorption spectroscopy (XAS) techniques. The results obtained by X-ray absorption near edge structure (XANES) spectra revealed that the mass fraction of PbO in MSWI fly ash was 57.9% (wt %) while PbCl2 and PbS were the dominant species in SFA. Extended X-ray absorption fine structure (EXAFS) data analysis indicated the atomic coordinates of Pb were proportional to the weights of PbCl2 and PbS, in good agreement with the XANES spectra. These findings highlight lead evaporation processes in the MSWI fly ash during heat treatment and provide a method for consistent speciation analysis of environmental samples using XAS.

Herpes simplex virus VP22 enhances adenovirus-mediated microdystrophin gene transfer to skeletal muscles in dystrophin-deficient (mdx) mice.

One of the obstacles to efficient vector-mediated gene therapy for Duchenne's muscular dystrophy (DMD) is its limited transduction efficiency. The VP22 tegument protein of herpes simplex virus type 1 (HSV-1) is able to cross biological membranes and translocate the VP22 fusion protein from transfected primary cells to surrounding cells and improve the outcome of gene transfer. To improve the efficiency of vector-mediated gene therapy and to investigate the utility of the intercellular trafficking properties of VP22-linked protein for the treatment for DMD, the recombinant adenoviruses Ad-VP22, Ad-MICDYS, and Ad-VP22-MICDYS were constructed and the VP22-mediated shuttle effect was evaluated both in vitro and in vivo. About 92 +/- 3.6% of cells were microdystrophin positive 48 hr postinfection with Ad-VP22-MICDYS. The number of centralized nuclei in Ad-VP22-MICDYS-transduced tibialis anterior (TA) muscle was significantly reduced, from 78 +/- 5.2 to 20 +/- 2.5%, by 2 weeks postinjection. By 2 months postinjection, the average number of microdystrophin-positive fibers in TA muscle injected with Ad-VP22-MICDYS was 2.2 times more than that of TA muscle injected with Ad-MICDYS. Ad-VP22-MICDYS led to significant recovery of force-producing capabilities in TA muscle. These results demonstrate that VP22 greatly augmented adenovirus-mediated microdystrophin delivery to C2C12 cells and to the skeletal muscles of dystrophin-deficient (mdx) mice. These results highlight the efficiency of VP22-mediated intercellular protein delivery for the potential therapy of DMD and suggest that VP22 may be a promising tool with which to enhance the efficacy of adenoviral gene transfer for somatic gene therapy of DMD.