In Situ Monitoring and Assessment of Ischemic Skin Flap by High-Frequency Ultrasound and Quantitative Parameters.

Skin flap surgery is a critical procedure for treating severe skin injury in which post-surgery lesions must well monitored and cared for noninvasively. In the present study, attempts using high-frequency ultrasound imaging, quantitative parameters, and statistical analysis were made to extensively assess variations in the skin flap. Experiments were arranged by incising the dorsal skin of rats to create a skin flap using the chamber model. Measurements, including photographs, 30 MHz ultrasound B-mode images, skin thickness, echogenicity, Nakagami statistics, and histological analysis of post-surgery skin flap, were performed. Photograph results showed that color variations in different parts of the skin flap may readily correspond to ischemic states of local tissues. Compared to post-surgery skin flap on day 7, both integrated backscatter (IB) and Nakagami parameter (m) of the distal part of tissues were increased, and those of the skin thickness were decreased. Overall, relative skin thickness, IB, and m of the distal part of post-surgery skin flap varied from 100 to 67%, -66 to -61 dB, and 0.48 to 0.36, respectively. These results demonstrate that this modality and quantitative parameters can be feasibly applied for long-term and in situ assessment of skin flap tissues.

Improved mycelia and polysaccharide production of Grifola frondosa by controlling morphology with microparticle Talc.

BACKGROUND: Mushroom showed pellet, clump and/or filamentous mycelial morphologies during submerged fermentation. Addition of microparticles including Talc (magnesium silicate), aluminum oxide and titanium oxide could control mycelial morphologies to improve mycelia growth and secondary metabolites production. Here, effect of microparticle Talc (45 µm) addition on the mycelial morphology, fermentation performance, monosaccharide compositions of polysaccharides and enzymes activities associated with polysaccharide synthesis in G. frondosa was well investigated to find a clue of the relationship between polysaccharide biosynthesis and morphological changes. RESULTS: Addition of Talc decreased the diameter of the pellets and increased the percentage of S-fraction mycelia. Talc gave the maximum mycelial biomass of 19.25 g/L and exo-polysaccharide of 3.12 g/L at 6.0 g/L of Talc, and mycelial polysaccharide of 0.24 g/g at 3.0 g/L of Talc. Talc altered the monosaccharide compositions/percentages in G. frondosa mycelial polysaccharide with highest mannose percentage of 62.76 % and lowest glucose percentage of 15.22 % followed with the corresponding changes of polysaccharide-synthesis associated enzymes including lowest UDP-glucose pyrophosphorylase (UGP) activity of 91.18 mU/mg and highest UDP-glucose dehydrogenase (UGDG) and GDP-mannose pyrophosphorylase (GMPPB) activities of 81.45 mU/mg and 93.15 mU/mg. CONCLUSION: Our findings revealed that the presence of Talc significantly changed the polysaccharide production and sugar compositions/percentages in mycelial and exo-polysaccharides by affecting mycelial morphology and polysaccharide-biosynthesis related enzymes activities of G. frondosa.

Simultaneous identification and quantification of canrenone and 11-α-hydroxy-canrenone by LC-MS and HPLC-UVD.

A procedure for simultaneous identification and quantification of canrenone and its biotransformed product 11-α-hydroxy-canrenone by high-performance liquid chromatography with ultraviolet detector (HPLC-UVD) and mass spectrometry (LC-MS) methods was proposed. The optimal determination variables on the HPLC-UVD or LC-MS coupled with a ZORBAX Eclipse XDB-C18 column (150 mm × 4.6 mm, 5 µm) were set as follows: detection wavelength of 280 nm, mobile phase of water and methanol gradient elution, temperature for the chromatographic column of 30°C, flow rate of mobile phase of 0.8 mL/min, sample injection volume of 5 µL, and elution time of 40 min. The MS conditions were set as follows: the flow rate of sheath gas, aux gas, and sweep gas were kept at 35 arb, 5 arb, and 0 arb, respectively. The temperature of capillary was held at 300°C, and capillary voltage was set at 30.00 V. Tube lens were performed at 100.00 V. The proposed method was validated by linearity (r² ≥ 0.9910), average recovery (94.93%, RSD1.21%), precision (RSD ≤ 1.31%), limit of detection, and limit of quantification (LOD 0.1~0.12 mg/L, LOQ 0.5~0.67 mg/L), which proved to be affordable for simultaneously determining canrenone and its bio-transformed product 11-α-hydroxy-canrenone.

Complete mitochondrial genome of an Asian longicorn beetle, Olenecamptus bilobus Fabricius (Coleoptera: Cerambycidae: Lamiinae).

Olenecamptus bilobus Fabricius is widely distributed in some parts of Southeast and East Asia whose larvae bore under the bark of at least eleven plant families. The complete mitochondria genome of O. bilobus was 15,262 bp in length, with 37 genes, including 12 protein-coding genes (PCGs), 23 tRNA genes (tRNAs), and 2 rRNA genes (rRNAs). The A + T content is 76.91%, showing strong AT skew. Phylogenetic analysis indicated that O. bilobus had a close relationship with Olenecamptus subobliteratus Pic.

Wafer-scale functional circuits based on two dimensional semiconductors with fabrication optimized by machine learning.

Triggered by the pioneering research on graphene, the family of two-dimensional layered materials (2DLMs) has been investigated for more than a decade, and appealing functionalities have been demonstrated. However, there are still challenges inhibiting high-quality growth and circuit-level integration, and results from previous studies are still far from complying with industrial standards. Here, we overcome these challenges by utilizing machine-learning (ML) algorithms to evaluate key process parameters that impact the electrical characteristics of MoS2 top-gated field-effect transistors (FETs). The wafer-scale fabrication processes are then guided by ML combined with grid searching to co-optimize device performance, including mobility, threshold voltage and subthreshold swing. A 62-level SPICE modeling was implemented for MoS2 FETs and further used to construct functional digital, analog, and photodetection circuits. Finally, we present wafer-scale test FET arrays and a 4-bit full adder employing industry-standard design flows and processes. Taken together, these results experimentally validate the application potential of ML-assisted fabrication optimization for beyond-silicon electronic materials.

VASP phosphorylation at serine239 regulates the effects of NO on smooth muscle cell invasion and contraction of collagen.

Nitric oxide triggers cGMP-dependent kinase-mediated phosphorylation of the actin regulator vasodilator-stimulated phosphoprotein (VASP) at residue serine239. The function of this phosphorylation for smooth muscle cell (SMC) adhesion, spreading, matrix contraction, and invasion is not well understood. We reconstituted VASP deficient SMC with wild-type VASP (wt-VASP) or VASP mutants that mimic "locked" serine239 phosphorylation (S239D-VASP) or "blocked" serine239 phosphorylation (S239A-VASP). Collagen gel contraction was reduced in S239D-VASP compared to S239A-VASP and wt-VASP expressing cells and nitric oxide (NO) stimulation decreased gel contraction of wt-VASP reconstituted SMC. Invasion of collagen was enhanced in S239D-VASP and NO-stimulated wild-type SMCs compared to S239A-VASP expressing cells. Expression of S239D-VASP impaired SMC attachment to collagen, reduced the number of membrane protrusions, and caused cell rounding compared to expression of S239A-VASP. Treatment of wt-VASP reconstituted SMCs with NO exerted similar effects as expression of S239D-VASP. As unstimulated cells were spreading on collagen S239A-VASP and wt-VASP localized to actin fibers whereas S239D-VASP was enriched in the cytosol. NO interferes with SMC invasion and contraction of collagen matrices. This requires phosphorylation of VASP on serine239, which reduces VASP binding to actin fibers. These findings support the conclusion that VASP phosphorylation at serine239 regulates cytoskeleton remodeling.

Three-dimensional discrete element simulation of the runaway vehicle deceleration process on the arrester bed of truck escape ramps.

Due to imperfect design norms and guidelines for China's truck escape ramp, previous studies have not been able to reflect the effect of wheel subsidence process on the deceleration of runaway vehicles. A discrete element method was used to establish an aggregate discrete element and a wheel discrete element. The three-dimensional discrete element model for an aggregate-wheel combination was established based on a particle flow code in three dimensions on a software platform using the "FISH" language. The microscopic parameters of the aggregate discrete element particles and wheel discrete element particles were calibrated using a simulated static triaxial compression test and real vehicle test data, respectively. Four sets of numerical simulation tests were designed for analyzing the influence of the aggregate diameter, grade of the arrester bed, truckload, and entry speed on the wheel subsidence depth and stopping distance of runaway vehicles. The results indicate that the smaller the aggregate diameter and entry speed and the greater the truckload and grade of the arrester bed, the more easily the wheel falls into the gravel aggregate, the better the deceleration effect, and the smaller the stopping distance. As the wheel subsidence depth increases, the speed at the unit stopping distance decreases more quickly. The maximum subsidence depth mainly depends on the truckload. The research results can provide a theoretical basis for the design of the arrester bed length and the thickness of the aggregate pavement in a truck escape ramp.

Mixotrophic Chlorella sp. UJ-3 cultivation in the typical anaerobic fermentation effluents.

The growth of mixotrophic Chlorella sp. UJ-3 cultivated in the three typical anaerobic fermentation effluents was investigated in this paper. The results showed that the microalgae grew best under intermediate light intensity for all the types of fermentation effluents. The butyrate type fermentation effluents induced the fastest growth rate for Chlorella sp. UJ-3, with a maximal cell concentration of 3.8×107 cells/mL. Under intermediate light intensity, the volatile fatty acids (VFAs) were almost depleted on the fifth day of the cultivation for all the three types of fermentation systems. The ratios of chlorophyll a/b were all increased for the three systems, indicating enhanced energy-capturing capability of the microalgae for photosynthesis after the VFAs were depleted. The highest lipid content was 25.4%dwt achieved in the butyrate type fermentation, and the fatty acid compositions were found to be considerably different for these three types of fermentation systems.

Production and characterization of a novel acidophilic and thermostable xylanase from Thermoascus aurantiacu.

The thermostable fungus, Thermoascus aurantiacus M-2, which produces a novel acidophilic and thermostable xylanase was isolated and identified based on its morphology and comparison of the internal transcribed spacer rDNA gene sequence. The culture conditions and components of medium were optimized for T. aurantiacus M-2 to produce xylanase. T. aurantiacus M-2 produced xylanase at a maximum level of 39.07 U/mL after 8-d fermentation at 45 °C in the optimized medium. The purified xylanase produced by T. aurantiacus M-2 has a relative molecular mass of approximately 31.0 kD. The characteristics of purified xylanase were investigated. The purified T. aurantiacus xylanase exhibited maximum activity at 75 °C and pH 5.0, and it was stable after treatment at a pH range from 2.0 to 10.0 or a temperature range from 30 °C to 80 °C for 2-h. Mn2+ and Ag+ enhanced xylanase activity to 120.0% and 119.6%, respectively, while Mn2+ had the highest inhibition ratio, with a residual activity of 20.7%. This study provided a foundation for scaled-up production and application of xylanase.

Induction of apoptosis in SGC-7901 cells by polysaccharide-peptide GFPS1b from the cultured mycelia of Grifola frondosa GF9801.

The biological function of GFPPS1b, a novel polysaccharide-peptide isolated from cultured mycelia of Grifola frondosa GF9801, was well investigated. GFPS1b has anti-tumor activity and can significantly inhibit the proliferation of SGC-7901 cells, whereas slightly influences the growth of human normal liver cell line L-02. When treated with GFPS1b, SGC-7901 cells showed typical apoptotic morphological features such as the loss of villus and appearance of apoptotic bodies on the cell surface, volume reduction, and chromatin condensation, by scanning electron microscopy (SEM) and fluorescent microscopy (Hoechst 33342). The results of flow cytometry analysis and annexin V-PI assay showed that the SGC-7901 cell cycle was arrested in the G(2)/M phase, the subdiploid peak of DNA characteristic of apoptotic was also observed, and the apoptosis ratio was about 15.08%. DNA isolated from SGC-7901 cells cultured with GFPS1b showed a typical DNA 'ladders' of apoptosis in agarose gel electrophoresis. Further investigation results showed that the apoptotic machinery of SGC-7901 induced by GFPS1b was associated with drop in mitochondrial trans-membrane potential, upregulation of Bax, downregulation of Bcl-2, and activation of caspase-3. Our finding suggests that GFPS1b could suppress SGC-7901 cell growth and reduce cell survival via arresting cell cycle and inducing apoptosis of tumor cells.

Muc4/sialomucin complex, the intramembrane ErbB2 ligand, induces specific phosphorylation of ErbB2 and enhances expression of p27(kip), but does not activate mitogen-activated kinase or protein kinaseB/Akt pathways.

Muc4/sialomucin complex (SMC) is a multifunctional glycoprotein complex which can repress apoptosis in transfected tumor cells. Its transmembrane subunit acts as an intramembrane ligand for the receptor tyrosine kinase ErbB2 to induce the phosphorylation of ErbB2 and, by acting synergistically with the ErbB3 ligand neuregulin, can potentiate the phosphorylation of ErbB2 and ErbB3. In the present study we show that Muc4/SMC alone robustly induces the phosphorylation of ErbB2 to enhance the tyrosine phosphate epitope (Tyr1248) recognized by anti-phospho-ErbB2. Although this tyrosine phosphorylation has been implicated in cell transformation, it does not activate any of the three mitogen-activated protein kinases (MAPKs) or protein kinase B/Akt of the phosphatidyl inositol 3-kinase pathway. Instead, Muc4/SMC expression induces up-regulation of the cell cycle inhibitor p27(kip), consistent with the expression of Muc4/SMC in differentiated, rather than proliferative, epithelial cells. Interestingly, a combination of Muc4/SMC and neuregulin down-regulate p27(kip) and activate protein kinase B/Akt. These observations suggest that Muc4/SMC acts as a regulator of differentiation by inducing a limited phosphorylation of ErbB2 and a modulator of proliferation when acting synergistically with neuregulin to induce a more extensive phosphorylation on both ErbB2 and ErbB3.

Sol-gel synthesis of luminescent InP nanocrystals embedded in silica glasses.

III-V semiconductor nanocrystals rarely exist as spherical inclusions inside glasses, due to difficulties during their preparation, such as high toxic reagents or fast oxidation under usual glass technology temperatures. In this letter a sol-gel method for synthesis of InP nanocrystals embedded in silica glasses was described. Gels were synthesized by hydrolysis of a complex solution of Si(OC2H5)4, InCl3.4H2O, and PO(OC2H5)3. Then, the gels were heated at 600 degrees C in the presence of H2 gas to form fine cubic InP crystallites. Raman spectrum showed an InP longitudinal-optic mode (342 cm(-1)) and a transverse-optic mode (303 cm(-1)). The size of InP nanocrystals was found to be from 2 to 8 nm in diameter by transmission electron microscopy. A strong photoluminescence with a peak at 856 nm was observed from InP nanocrystals embedded in silica glasses. The results suggest that it might be possible to synthesize other III-V semiconductor nanocrystals embedded in silica glasses through the sol-gel process.

Sol-gel synthesis and photoluminescence of III-V semiconductor InAs nanocrystals embedded in silica glasses.

III-V semiconductor nanocrystals rarely exist as spherical inclusions inside glasses, due to the difficulties during their preparation, such as high toxic reagents or fast oxidation under usual glass technology temperatures. In this paper a sol-gel method for synthesis of InAs nanocrystals embedded in silica glasses was described. Gels were synthesized by the hydrolysis of a complex solution of Si(OC2H5)4, InCl3 x 4H2O, and As2O3. The gels were then heated at 200-450 degrees C in the presence of H2 gas to form fine cubic InAs crystallites. The X-ray diffraction patterns showed four strong peaks from InAs. The Raman spectrum showed InAs longitudinal-optic (233 cm(-1)) and transverseoptic modes (215 cm-(-1)). The size of InAs nanocrystals was found to be from 5 to 30 nm in diameter by transmission electron microscopy. A strong room temperature photoluminescence with peaks at 601 and 697 nm was observed from InAs nanocrystals embedded in silica glasses. The results suggest that it might be possible to synthesize other III-V semiconductor nanocrystals embedded in silica glasses through the sol-gel process.

Molecular determinants of cGMP binding to chicken cone photoreceptor phosphodiesterase.

Structural studies on photoreceptor phosphodiesterases type 6 (PDE6s) have been hampered by an inability to express and purify substantial amounts of enzyme. Here we describe bacterial expression and characterization of the chicken cone PDE6 regulatory GAF-A and GAF-B domains. High affinity cGMP binding was found only for GAF-A as predicted from sequence alignments with the GAF domains of PDE2 and PDE5. A homology model of the GAF-A domain of chicken cone PDE6 based on the crystal structure of mouse PDE2A GAF-B was used to identify residues likely to make contact with cGMP. Alanine mutagenesis of 4 of these residues (F123A, D169A, T172A, and T176A) showed that each was absolutely required for cGMP binding. Three of these residues map to the H4 helical structure of the GAF-A domain indicating this region as a key structural component for cGMP binding. Mutagenesis of another residue, S97A, decreased cGMP binding affinity 5-fold. Finally mutagenesis of Glu-124 indicated that it is responsible for part but not all of the high specificity for cGMP binding to PDE6 GAF-A. Since little data is available on the properties of the chicken cone PDE6 holoenzyme, we also characterized the native PDEs of chicken retina. Two histone-activated PDE6 peaks were separated by ion exchange chromatography and identified by mass spectrometry as cone and rod photoreceptor PDE6s, respectively. Both of these PDEs had cGMP binding and kinetic properties similar to their corresponding bovine photoreceptor PDEs. Moreover the cGMP binding properties of chicken cone PDE6 holoenzyme were very similar to those of the bacterially expressed individual GAF-A or GAF-A/B domains.