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Citrus greening (Huanglongbing, HLB) has seriously threatened the healthy development of citrus industry in recent years, and it is great significance to diagnose citrus HLB timely and accurately. Hyperspectral technology has the characteristics of spectral analysis and image processing, which has shown great advantages in plant disease detection. The visual discrimination methods of citrus HLB based on features of images combined with hyperspectral imaging technology were discussed. Five types of citrus leaves, including the mild HLB, moderate HLB, serious HLB, malnourished and normal were studied and polymerase chain reaction (PCR) test was used to verify visual division. Effective spectral variables were selected by successive projections algorithm (SPA) and features of images were extracted by principal component analysis (PCA). Texture features of images based on grayscale co-occurrence matrix (GLCM) were used to develop the partial least squares discriminant analysis (PLS-DA) models. The influence of the number of textures on the models was discussed, and the effect of discrimination model was the best when a total of 36 variables which were obtained from 4 independent texture features were as input, which resulted in greater misjudgment rate of 3.12%, higher correlation coefficient for prediction (RP) of 0.98, lower root mean square error for prediction (RMSEP) of 0.32, and the number of principal component factors (PCs) of 16. The results highlighted that the image texture features based on GLCM combined with the PLS-DA models could realize the identification of citrus HLB, and provide the important reference value for the visual discrimination research of HLB.

Investigations were carried out to develop terahertz (THz) techniques combined with machine learning methods for rapidly measuring benzoic acid (BA) additive in wheat flour. The absorption coefficient had a significant characteristic peak at 1.94 THz, which usually varies with the increase of BA additive concentrations. Correlation analysis was initiated to choose a frequency pair of 1.94 and 1.86 THz that improved the predictive ability of multiple linear regressions (MLR) model. Then, quantitative determination models were modeled between BA concentrations and THz absorption coefficients using partial least square (PLS) regression and least square support vector machine (LS-SVM). Compared with MLR and PLS models, the LS-SVM model could be regarded as an effective tool for quality control of wheat flour with a correlation coefficient of prediction (Rp) of 0.994 and root mean square error of prediction (RMSEP) of 0.12%. The results suggest that combining THz spectroscopy with LS-SVM has the potential to quantitatively analyze BA additive in wheat flour.

With model uncertainties and input saturation, a novel control method is developed to steer an underactuated autonomous underwater vehicle that realizes the following of the planned path in three-dimensional (3D) space. Firstly, Serret–Frenet frame is applied as virtual target, and the path following errors model in 3D is built. Secondly, the control method which includes kinematic controller and dynamic controller was presented based on cascade control strategy. The kinematic controller, which is responsible for generating a series of constrained velocity signals, is designed based on model predictive control. The adaptive radial basis function neural network is used to estimate the model uncertainty caused by hydrodynamic parameters. Moreover, sliding mode control technology is applied in the design of dynamic controller to improve its robustness. Then, the control effect is compared with that of LOS guidance law and PID controller by simulation experiment. The comparison results show that the proposed algorithm can improve path following effect and reduce input saturation.

Saponins form a group of plant-produced glycosides with potential as biopesticide ingredients. The environmental fate of saponins has never been fully investigated. In the present study, we use QS-18, a specific saponin from Quillaja saponaria as an example, to quantify hydrolysis under different conditions of pH, temperature and water chemical composition. Saponin hydrolysis in buffer solutions was base-catalyzed and followed first-order kinetics. Thus, hydrolysis was slow at pH 5.1 with a half-life of 330 ± 220 d (26 °C), which increases to 0.06 ± 0.01 d at pH 10.0. Hydrolysis rates were highly sensitive to temperature with an activation energy of 56.9 ± 14.2 kJ mol-1 at pH 7.2. In strong contrast, hydrolysis in lake waters (pH 6.4-8.2) produced different patterns with a fast initial dissipation of 25 to 60% of the added saponin within the first five hours, followed by an extremely slow reaction with 25 to 75% unreacted saponin left after reaction times longer than 120 h. The fast dissipation followed by slow hydrolysis in lake water was hypothesized to be attributed to sorption and/or flocculation of saponins by inorganic nanoparticles and/or solutes in the lake water followed by inactivation of hydrolysis due to the sorption/flocculation. The present study demonstrates that saponins may hydrolyze slowly under acidic and cold conditions. In addition, it demonstrates that dissipation kinetics in natural waters may deviate substantially from the kinetics predicted based on laboratory experiments with "clean" buffered solutions. This emphasizes the need for a deeper understanding of the processes affecting the dissipation kinetics of potential toxins under natural conditions, as fate models based on laboratory derived kinetic data may be seriously flawed.

Previous cDNA microarray results showed that MYH9 gene expression levels are increased in TGF-β1-stimulated lung fibroblast. Recently, our proteomic results revealed that the expression levels of MYH9 protein are notably upregulated in lung tissues of bleomycin-treated rats. However, whether MYH9 plays a critical role in the differentiation of fibroblast remains unclear. Herein, we demonstrated that TGF-β1 increased MYH9 expression, and siRNA-mediated knockdown of MYH9 and pharmacological inhibition of MYH9 ATPase activity remarkably repressed TGF-β1-induced lung fibroblast-to-myofibroblast differentiation. TGF-β1-stimulated MYH9 induction might be via ALK5/Smad2/3 pathway but not through noncanonical pathways, including p38 mitogen-activated kinase, and Akt pathways in lung fibroblasts. Our results showed that MYH9 inhibition suppressed TGF-β1-induced lung fibroblast-to-myofibroblast differentiation, which provides valuable information for illuminating the pathological mechanisms of lung fibroblast differentiation, and gives clues for finding new potential target for pulmonary fibrosis treatment.

Although we have reported that baicalein ameliorated bleomycin-induced pulmonary fibrosis in rats and inhibited fibroblast-to-myofibroblast differentiation, the mechanisms of the capability of baicalein to suppress the production of type I collagen in fibroblasts remains unclear. Here, we showed that baicalein suppressed transforming growth factor β1 (TGF β1)-stimulated the production of type I collagen in lung fibroblast MRC-5 cells. By applying SILAC-based proteomic technology, 158 proteins were identified as baicalein-modulated proteins in TGF β1-stimulated the accumulation of type I collagen in MRC-5 cells. Our proteomic and biochemical analysis demonstrated that baicalein decreased the expression levels of connective tissue growth factor (CTGF) in TGF β1-stimulated MRC-5 cells. In addition, CTGF overexpression elevated the levels of type I collagen in baicalein-treated fibroblasts. Moreover, our results demonstrated that baicalein-downregulated CTGF expression might be related with the decrease of Smad2 phosphorylation, but not SP1. This work not only linked CTGF to TGF β1-stimulated the production of type I collagen in its attribution to the effects of baicalein, but also might provide valuable information for enhancing the knowledge of the pharmacological inhibition of collagen production, which might represent a promising strategy for the treatment of pulmonary fibrosis.

Blackheart and soluble solids content (SSC) are critical parameters of pear for quality assessment and export trade guarantee. A non-destructive detection method was developed for measuring blackheart pear and SSC. Three discriminate analysis models, such as principal component analysis (PCA), correlation analysis and partial least square discriminate analysis (PLS-DA), were developed to identify black heart samples. Compared with PCA and correlation analysis, PLS-DA achieved the highest classification accuracy of 100% for identifying blackheart pears. Three sample sets of healthy samples, black heart ones, and their combination were applied to assess the performance of PLS model, and PLS model calibrated with healthy pears yielded better with root-mean-square errors of prediction 0.45°Brix. The overall results suggested that it was feasible to detect blackheart and SSC of pears simultaneously by online visible–NIR spectroscopy.

Saponin-rich plant extracts contain bioactive natural compounds and have many applications, e.g. as biopesticides and biosurfactants. The composition of saponin-rich plant extracts is very diverse, making environmental monitoring difficult. In this study various ecotoxicity data as well as exposure data have been collected to explore which compounds in the plant extract are relevant as plant protection agents and furthermore to clarify which compounds may cause undesired side-effects due to their toxicity. Hence, we quantified the toxicity of different fractions (saponins/non-saponins) in the plant extracts on the aquatic crustacean Daphnia magna and zebrafish (Danio rerio) embryos. In addition, we tested the toxicity changes during saponin degradation as well. The results confirm that saponins are responsible for the majority of toxicity (85.1-93.6%) of Quillaja saponaria extract. We, therefore, suggest saponins to be the main target of saponin-rich plant extracts, for instance in the saponin-based biopesticide regulation. Furthermore, we suggest that an abundant saponin fraction, QS-18 from Q. saponaria, can be a key monitoring target to represent the environmental concentration of the saponins, as it contributes with 26% and 61% of the joint toxicity to D. magna and D. rerio, respectively out of the total saponins. The degradation products of saponins are 3-7 times less toxic than the parent compound; therefore the focus should be mainly on the parent compounds.

Saponin-rich extracts from Quillaja saponaria and Chenopodium quinoa have been registered by US EPA as active ingredients in biopesticides, and extract from tea seed powder, Camellia oleifera has been proposed for biocidal use. If saponin-rich biopesticides are efficient against pests, they are most likely also bioactive in the aquatic environment against non-target organisms. The aim of this study was to conduct an effect assessment of saponin-rich plant extracts by using species sensitivity distributions based on acute toxicity tests. The maximal concentrations protecting 95% of the aquatic species (HC5) of saponins extracted from quillaja bark, tea seed coat and quinoa seed coat were 2.91 ± 1.00, 0.22 ± 0.11 and 22.9 ± 5.84 mg/L, respectively. The 100-fold difference in toxicity between the saponin-rich extracts from different plant species, indicate that saponin toxicity depends on the species it origins from, making “read-across” between saponins a dubious exercise. In addition, the predicted environmental concentrations of different saponins are close to or higher than their water quality standard, which means that the extracts might pose a risk to the aquatic environment if not used cautiously.

Protein post-translational modification by ubiquitin-fold modifier 1, UFM1, regulates many biological processes such as response to endoplasmic reticulum stress and regulation of tumor progression. A recent study has indicated that the UFM1-binding and PCI domain-containing protein 1 (UFBP1) is required for the conjugation of UFM1 to a substrate. However, other biological functions of UFBP1 have not been explored. Here, we use immunoprecipitation and label-free quantitative proteomics to identify UFBP1-interacting proteins in a mammalian cell line. About 80 potential interacting proteins are obtained from MS analyses of three biological replicates. Bioinformatics analyses of these proteins suggest that UFBP1 may participate in the regulation of protein folding, stability, and trafficking. Biochemical experiments discover that UFBP1 expression downregulates the protein level and reduces the stability of several of its interacting proteins, while UFBP1 knockdown increases their protein levels. Protein synthesis inhibition and proteasomal inhibition experiments reveal that UFBP1 promotes their ubiquitination and degradation. Experiments using a model UFBP1-interacting protein ANT3 demonstrate that UFBP1 enhances the interaction between ANT3 and its E3 ligase and thus promotes its ubiquitination and degradation. Our work elucidates a novel molecular mechanism by which UFBP1 regulates protein ubiquitination and degradation.

The liver cancer has become the sixth most common cancer and the second leading cause of cancer deaths worldwide. The incidence of liver cancer has characteristics of the geographical distribution, which vary from region to region. Based on the data from Cancer Incidence in Five Continents, this study analyzed the spatial distribution of liver cancer globally and explored the effect of meteorological factors of surface temperature and wind field on incidence of liver cancer, to find the geographical distribution characteristics of liver cancer and the impact of geographical environmental factors on the incidence of liver cancer, which will be of great significance to the epidemiological study of liver cancer and the prevention of liver cancer. The results show that the humid and hot oceanic climate may be conducive to the development of liver cancer. The environmental factors play an important role in the occurrence of liver cancer.

In the current study, a specially designed automated nanoflow liquid chromatography coupled with tandem mass spectrometer (μLC-MS/MS) system for phosphopeptide analysis was reported. The system was established by applying a strong anion exchange (SAX) trap column. At the beginning of the analysis, phosphopeptides were loaded onto SAX trap column at high flow rate. Next, the retained phosphopeptides were eluted onto a C18 analytical capillary column by an acidic buffer with high ionic strength. At last, its performance was evaluated by analyzing the phosphopeptides enriched from the tryptic digest of mouse liver lysate. Compared with conventional automated μLC-MS/MS system using C18 trap column, much more phosphopeptides could be identified by the SAX trap column system. This system was fully automated and had promising application in high throughput phosphoproteomic analysis.

Investigations were initiated to develop terahertz (THz) techniques associated with machine learning methods of generalized regression neural network (GRNN) and back-propagation neural network (BPNN) to rapidly measure benzoic acid (BA) content in wheat flour. The absorption coefficient exhibited a maximum absorption peak at 1.94 THz, which generally increased with the content of BA additive. THz spectra were transformed into orthogonal principal component analysis (PCA) scores as the input vectors of GRNN and BPNN models. The best GRNN model was achieved with three PCA scores and spread value of 0.2. Compared with the BPNN model, GRNN model to powder samples could be considered very successful for quality control of wheat flour with a correlation coefficient of prediction ( r p ) of 0.85 and root mean square error of prediction of 0.10%. The results suggest that THz technique association with GRNN has a significant potential to quantitatively analyse BA additive in wheat flour.

A 187 fs pulse laser with 2.3 W of output power at 1560 nm is built for second-harmonic generation (SHG), which is the highest average power output reported from an all-polarization-maintaining single-mode Er-doped chirped pulse amplification system. By subsequently using an MgO-doped periodically poled lithium niobate (MgO:PPLN) crystal as frequency doubler, a 183 fs pulse laser with 1.1 W output power at 778 nm is generated. Benefitting from a polarization-stable fundamental laser input and the temperature control of the PPLN crystal, the power fluctuation of the frequency doubled laser is less than 0.5% during 24 h. To the best of our knowledge, the 1.1 W output at 780 nm band is also the highest average power generated from a frequency doubled femtosecond Er-doped fiber laser.

We present a single-frequency thulium-doped fiber laser (TDFL) with a narrow linewidth of 20 kHz. Stable single-longitudinal-mode (SLM) lasing operation at 1957 nm is achieved using a segment of un-pumped polarization-maintaining thulium-doped fiber (PM-TDF) as an ultra-narrow bandwidth filter. A high optical signal-to-noise ratio (OSNR) of over 60 dB is obtained and a high power of over 400 mW is achieved with a high slope-efficiency (~45.8%) thulium-doped fiber amplifier (TDFA).

We present a linearly polarized single-longitudinal-mode (SLM) ytterbium-doped fiber laser with a polarization extinction ratio (PER) of over 35 dB and a narrow linewidth of less than 4.5 kHz. The very high PER is obtained by utilizing the polarization evolution effect at the optical fiber and the high-performance polarizing beam splitter. The SLM is achieved by using a segment of polarization-maintaining ytterbium-doped fiber as a narrowband filter. In addition, a high optical signal-to-noise ratio of 50 dB and a good slope efficiency of 60.5% are achieved. Using an ytterbium-doped fiber amplifier, a linearly polarized SLM laser system with a high power of over 2.5 W is demonstrated at 1064 nm.

Peaches are sensitive to bruising from mechanical impact and compression. Peaches affected by bruises will tend towards fermentation, decay or mildew and infect other non-bruised ones after damage occurrence. A moderate amount of bruising is a barrier to purchase desire of consumers instead of price. For the early bruised peaches, softening and browning does not appear immediately (within 2 days), and it is difficult to distinguish the problem of fruit bruises. The hypersecretion imaging device was used to collect the data of normal peaches and the data of 12, 24, 36 and 48 h after the encounter, and the spectral characteristics of these five types of peaches were analyzed to establish the PCA classification model of physiological indexes of peaches after the collision. Extract the spectrum and image features of the discolored area and the normal area. Secondly, divide the collection of image data sets into the modelling set and the prediction set at a ratio of 3:1. Input the spectrum and image feature to establish a time discrimination model for peach bruises. The results show that it is based on the PLS-DA algorithm When the input variable is a spectral feature, the classification accuracy rates at 12, 24, 36, and 48 h after bruising are 96.67%, 96.67%, 93.33%, and 83.33%, respectively, and the correlation coefficient (Rc) of the modelling set reaches 0.928; Based on LS -SVM algorithm, when the input variable is a spectral feature, the kernel function RBF_Kernel classifies correctly to 80%, 96.67%, 100%, 100%. This study shows that the spectral feature of hypersecretion technology can identify early bruises of peaches, which are online testing provide a theoretical basis for predicting short-term bumps of peaches.

We demonstrate a mid-infrared (MIR) wavelength-tunable high-power single-longitudinal-mode (SLM) fiber laser using a compound cavity structure. The compound cavity consists of an Er-doped ZBLAN fiber perpendicularly cleaved at two-ends and an external cavity formed by a fiber end-face and a narrow bandwidth ruled reflective diffraction grating. SLM operation is achieved through the combined effects of the narrow bandwidth grating and the compound cavity. By rotating the grating, the laser wavelength can be continuously tuned over 100 nm from 2705 nm to 2806 nm with the SLM operation maintained. High output power up to 450 mW is achieved under the SLM regime.

Stone fruits have a single-core, and the core volume is large, located in the center of the fruit. In the process of transmission and reflection spectroscopic detection, the influence of fruit core on light cannot be ignored when light passes through flesh. Analyzing the difference in optical characteristics between stone fruits and citrus fruits tissues can provide a theoretical basis for the design of a reasonable optical detection device. It is of great significance for the optical detection of fruit quality. In this paper, a self-developed near-infrared diffuse reflection optical fibre probe with multi-point emission and multi-point reception is used to collect the diffuse reflection spectra of peach and navel orange, and compare the spectral characteristics of the two fruits. The spectral energy of peach is about twice as strong as the spectral energy of navel orange between 600 nm and 900 nm. Preprocess the spectral data with preprocessing methods to reduce the influence of noise and stray light. Compare the partial least squares (PLS) regression models of peach and navel orange. Experiments show that the results of using the baseline correction preprocessing method are all the best. The prediction correlation coefficient (Rp) of the peach is better than that of the navel orange and the root mean square error of the prediction (RMSEP) is also lower. The Rp of peach reached 0.92, and RMSEP was 0.32°Brix. It turned out that after light enters the fruit tissues, part of the light will be reflected out of the fruit when it encounters the core of the fruit. Therefore, in the diffuse reflection detection mode, stone fruits can receive more light and the spectral energy will be stronger than that of citrus fruits.

In order to study the changes of internal quality of peach (juicy peach) during storage and further discuss the feasibility of predicting the quality of peach through visible /near infrared spectroscopy (VIS/NIR). Firstly, the spectra of peach in two storage conditions were collected with the fruit quality dynamic detection equipment, and its soluble solid content (SSC), firmness and weight were measured. Then, partial least squares discrimination analysis (PLS-DA) was used to develop the classification model of peach, and the accuracy was higher than 90 %. Finally, partial least squares (PLS) regression combined with preprocessing methods was used to develop the models of SSC and firmness. Uninformative variable elimination (UVE) and competitive adaptive reweighted sampling (CARS) were also used to optimize the model to obtain the optimal prediction models. The results showed that CARS-PLS models of peach at the room temperature were the optimal, with prediction correlation coefficient (Rp) at 0.819, and the root mean square error of prediction (RMSEP) at 0.841°Brix. The Rp and RMSEP of firmness were 0.811 and 0.912 N, respectively. In cold storage, the PLS model of SSC processed by CARS was the optimal with the Rp of 0.828 and RMSEP of 0.816°Brix. The Rp and RMSEP of the optimal firmness model were 0.785 and 1.188 N, respectively. This study on the quality of peach can better analyze the quality change of peach during storage, and has certain value for guiding the storage of peach and subsequent related research.

As a locomotive fuel, gasoline has become one of the main consumer products of Chinese car owners. Effective identification of gasoline adulteration is of great significance to personal safety, car maintenance and environmental protection. This study used mid-infrared and Raman spectroscopy to quantitatively analyze kerosene adulteration in gasoline. The content of kerosene in gasoline by mid-infrared and Raman spectroscopy was established by partial least squares (PLS), extreme learning machine (ELM) and random forest model prediction. Analyze the effects of Savitzky-Golay (S-G), Multiple scatter correction (MSC), standard normal variate (SNV), first-order, second-order and two pre-processing methods, and calculate the prediction results of the best pre-processing method under the model. The mid-infrared PLS model combined with SG superposition SNV preprocessing method, ELM model combined with SNV preprocessing method, random forest model combined with SG superposition 1st preprocessing method, the prediction correlation coefficients are 0.9828, 0.9374, 0.9817, respectively, And the root mean square error is 0.7878, 0.3606, 0.2175. The results show that the mid-infrared and Raman vibration spectroscopy methods can effectively detect the kerosene content in gasoline, while the mid-infrared PLS model combined with SG superposition SNV pretreatment method has the best effect The research is carried out for gas stations, production enterprises and supervision departments.

Fibroblast-to-myofibroblast differentiation is one of the most important characteristics of pulmonary fibrosis, and screening natural compounds targeting fibroblast differentiation is always a promising approach to discover drug candidates for treatment of pulmonary fibrosis. Trehalose reportedly has many potential medical applications, especially in treating neurodegeneration diseases. However, it remains unclear whether trehalose suppresses lung fibroblast differentiation. In this work, we found that trehalose decreased the expression levels of α-smooth muscle actin (α-SMA) following the induction of transforming growth factor β1 (TGF-β1) in pretreatment, co-treatment, and post-treatment groups. Trehalose also reduced the production of type I collagen, lung fibroblast-containing gel contractility and cell filament formation in TGF-β1-stimulated MRC-5 cells. Although trehalose is a known autophagy inducer, our results showed that its suppressive effect on fibroblast differentiation was not via trehalose-induced autophagy. And it did not affect canonical TGFβ/Smad2/3 pathway. By applying proteomic profiling technology, we demonstrated that the downregulation of β-catenin was involved in the trehalose-repressive action on fibroblast differentiation. The β-catenin agonist, SKL2001, reversed the suppressive effect of trehalose on fibroblast differentiation. Overall, these experiments demonstrated that trehalose suppressed fibroblast differentiation via the downregulation of β-catenin, but not through canonical autophagy and TGFβ/Smad2/3 pathway, which is not only a novel understanding of trehalose, but also quite helpful for in vivo research of trehalose on pulmonary fibrosis in future.

Fibroblast-to-myofibroblast differentiation is a highly important pathological characteristic of pulmonary fibrosis. In this study, we aimed to investigate the effects and mechanisms of baicalein on the differentiation of human lung fibroblasts. Baicalein reduced the levels of α-smooth muscle actin (α-SMA) mRNA and protein expression in TGF-β1-treated human lung fibroblasts. It also decreased the contents of collagen type I and fibronectin in time- and dose-dependent manners, and retarded TGF-β1-stimulated α-SMA filament formation. Baicalein diminished the expression of miR-21, and miR-21 mimics partially antagonized the effects of baicalein. Additionally, Baicalein inhibited the miR-21 transcriptor STAT3 activity but not AP-1 activity. Moreover, the expression of Spry 1 protein, a miR-21 known target, was improved by baicalein treatment, but the level of Smurf2 protein, another miR-21 target, was not interfered. Collectively, these results demonstrated that baicalein can attenuate TGF-β1-induced human lung fibroblast differentiation by inhibiting the miR-21 expression.

A wavelength-tunable all-fiber mode-locked erbium-doped fiber laser has been proposed and realized by using a supermode interference filter (SMIF). The SMIF is fabricated by splicing a segment of seven-core fiber (SCF) to two standard single-mode fibers. Since two supermodes of the propagating light are excited in the SCF, the transmission spectrum of the SMIF shows a clean broadband comb-shape characteristic. By bending the SMIF in the proposed mode-locked laser, the output spectrum can be continuously tuned in a wavelength range up to 22 nm while keeping mode-locking operation. The self-starting laser produces 230 fs pulses with a spectral width of 14 nm.

Chronic inflammation is associated with multiple human disorders, such as rheumatoid arthritis, metabolic diseases, and neurodegenerative diseases. Therefore, alleviation of inflammation induced by environmental stimuli is important for disease prevention or treatment. Cereblon (CRBN) functions as a substrate receptor of the cullin-4 RING E3 ligase to mediate protein ubiquitination and degradation. Although it has been reported that CRBN reduces the inflammatory response through its nonenzymatic function, its role as a substrate receptor of the E3 ligase is not explored in mediating this process. Here we used a quantitative proteomics approach to find that the major component of the activator protein 1 (AP-1) complex, c-Jun, is significantly down-regulated upon CRBN expression. Biochemical approaches further discover that CRBN interacts and partially colocalizes with c-Jun and promotes the formation of Lys(48)-linked polyubiquitin chains on c-Jun, enhancing c-Jun degradation. We further reveal that CRBN attenuates the transcriptional activity of the AP-1 complex and reduces the mRNA expression and protein level of several pro-inflammatory cytokines. Moreover, flow cytometry analyses show that CRBN attenuates lipopolysaccharide-induced apoptosis in differentiated THP-1 cells. Through genetic manipulation and pharmacological inhibition, we uncover a new molecular mechanism by which CRBN regulates the inflammatory response and apoptosis induced by lipopolysaccharide. Our work and previous studies demonstrate that CRBN suppresses the inflammatory response by promoting or inhibiting the ubiquitination of two key molecules at different levels of the inflammatory cascade through its enzymatic function as a substrate receptor and its nonenzymatic function as a protein binding partner.

The immunomodulatory drug (IMiD) thalidomide and its structural analogs lenalidomide and pomalidomide are highly effective in treating clinical indications. Thalidomide binds to cereblon (CRBN), a substrate receptor of the cullin-4 really interesting new gene (RING) E3 ligase complex. Here, we examine the effect of thalidomide and its analogs on CRBN ubiquitination and its functions in human cell lines. We find that the ubiquitin modification of CRBN includes K48-linked polyubiquitin chains and that thalidomide blocks the formation of CRBN-ubiquitin conjugates. Furthermore, we show that ubiquitinated CRBN is targeted for proteasomal degradation. Treatment of human myeloma cell lines such as MM1.S, OPM2, and U266 with thalidomide (100 μM) and its structural analog lenalidomide (10 μM) results in stabilization of CRBN and elevation of CRBN protein levels. This in turn leads to the reduced level of CRBN target proteins and enhances the sensitivity of human multiple myeloma cells to IMiDs. Our results reveal a novel mechanism by which thalidomide and its analogs modulate the CRBN function in cells. Through inhibition of CRBN ubiquitination, thalidomide and its analogs allow CRBN to accumulate, leading to the increased cullin-4 RING E3 ligase-mediated degradation of target proteins.—Liu, Y., Huang, X., He, X., Zhou, Y., Jiang, X., Chen-Kiang, S., Jaffrey, S. R., Xu, G. A novel effect of thalidomide and its analogs: suppression of cereblon ubiquitination enhances ubiquitin ligase function.

Amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD) are the two common neurodegenerative diseases that have been associated with the GGGGCC·GGCCCC repeat RNA expansion in a noncoding region of C9orf72. It has been previously reported that unconventional repeat-associated non-ATG (RAN) translation of GGGGCC·GGCCCC repeats produces five types of dipeptide-repeat proteins (referred to as RAN proteins): poly-glycine-alanine (GA), poly-glycine-proline (GP), poly-glycine-arginine (GR), poly-proline-arginine (PR) and poly-proline-alanine (PA). Although protein aggregates of RAN proteins have been found in patients, it is unclear whether RAN protein aggregation induces neurotoxicity. In the present study, we aimed to understand the biological properties of all five types of RAN proteins. Surprisingly, our results showed that none of these RAN proteins was aggregate-prone in our cellular model and that the turnover of these RAN proteins was not affected by the ubiquitin-proteasome system or autophagy. Moreover, poly-GR and poly-PR, but not poly-GA, poly-GP or poly-PA, localized to the nucleolus and induced the translocation of the key nucleolar component nucleophosmin, leading to nucleolar stress and cell death. This poly-GR- and poly-PR-mediated defect in nucleolar function was associated with the suppression of ribosomal RNA synthesis and the impairment of stress granule formation. Taken together, the results of the present study suggest a simple model of the molecular mechanisms underlying RAN translation-mediated cytotoxicity in C9orf72-linked ALS/FTD in which nucleolar stress, but not protein aggregation, is the primary contributor to C9orf72-linked neurodegeneration.

We reported a novel fiber sensor based on two cascaded single mode fiber corners, which has a high sensitivity for refractive index (RI) sensing and a low temperature crosstalk. RI sensing was experimentally demonstrated with an average sensitivity of 235 nm/RI unit in a RI range from 1.333 to 1.418, which is relatively high among the previously reported similar fiber sensors (e.g., it is about 10 times higher than that of the normal two or three-cascaded-taper based Mach–Zehnder interferometer). The proposed RI sensor was fabricated by fiber fusion splicer, where the fiber corner was fabricated by splicing the two overlapping single mode fibers, which ensures that the proposed RI sensor is compact and strong. © 2014 Wiley Periodicals, Inc. Microwave Opt Technol Lett 56:2642–2645, 2014

Electrocoagulation and electroflotation are effective technologies for restaurant wastewater treatment, especially for the removal of oil and grease. The response surface methodology was used to establish a model of restaurant wastewater treatment using electrocoagulation and electroflotation. The model provides the optimum operation conditions. In addition, the contributions of electrocoagulation and electroflotation to the removal of oil and grease were determined under different conditions. The optimum operation conditions are an inter-electrode distance of 3.6 cm, a reaction time of 34 min, and a current density of 43 A/m2. The removal efficiency of oil and grease was above 95% under such conditions. When wastewater conductivity was less than 3,000 μs/cm, electrocoagulation played the dominant role, and the contribution rates ranged from 68.1 to 72.5%. When the wastewater conductivity was above 3,500 μs/cm, electroflotation played the dominant role, and the contribution rates ranged from 51.6 ...

We propose, fabricate, and demonstrate a compact fiber-tip Fabry-Perot interferometer of microcavity for high-temperature sensing. The microcavity which can be used for temperature or pressure sensing is fabricated by using arc discharge at the end of a multimode fiber, which is processed with chemical etching. Advantages of the sensor based on the fiber-tip Fabry-Perot interferometer are small size, easy fabrication, low cost and may have potential applications in space-limited and high-temperature environment.

A kind of novel fiber, comprising two fiber cores which are suspended in air inside the outer cladding via a central thin membrane, is proposed for optical switching application. When a hydrostatic pressure applied on the optical fiber, the pressure-induced refractive index change of the two fiber cores will contribute to the periodical change of the intensity of guided light in the fiber core. The mode coupling of two cores under different hydrostatic pressure and influences of each structure parameter of the proposed fiber on the switching pressure have been numerically investigated.

A Fabry-Perot interferometer sensor based on a fiber-tip bubble-structure micro-cavity is proposed, fabricated, and demonstrated for transverse load sensing. The micro-cavity is fabricated by using arc discharge at the end of a multimode fiber which has been processed with chemical etching. A transverse load sensitivity of 3.64 nm/N and a relative low temperature sensitivity of about 2 pm/ ◦ C are experimentally demonstrated for the proposed fiber-tip bubble-structure micro-cavity sensor. The sensor has the advantages of low-cost, ease of fabrication and compact size, which make it a promising candidate for transverse load sensing in harsh environments.

Currently, there is no satisfactory treatment for pulmonary fibrosis, and effective agents urgently need to be developed. The aim of the present study was to investigate the effects of baicalein on bleomycin-induced pulmonary fibrosis, and the novel mechanisms involved in the anti-fibrosis effects. Pulmonary fibrosis was induced by a single intratracheal instillation of 5 mg/kg bleomycin. Two bleomycin-treated groups were orally administered daily with 50 and 100 mg/kg of baicalein from day 1 to 28. The results showed baicalein decreased hydroxyproline content and α-SMA levels and increased lung index. Histopathological examinations demonstrated baicalein could obviously lower the degree of alveolitis and lung fibrosis. The total antioxidant capacity in bleomycin-treated rats with baicalein was also remarkably higher than in those without baicalein. Baicalein remarkably decreased miR-21 levels and inhibited the increased expression of TGF-β1 and p-Smad-2/3 in bleomycin-treated rats. Baicalein can attenuate bleomycin-induced pulmonary fibrosis. The attenuation is partly achieved by improving antioxidant activity, alleviating inflammation, repressing miR-21, and inhibiting TGF-β/Smad signaling.

A nonsense mutation in cereblon (CRBN) causes autosomal recessive nonsyndromic mental retardation. Cereblon is a substrate receptor for the Cullin-RING E3 ligase complex and couples the ubiquitin ligase to specific ubiquitination targets. The CRBN nonsense mutation (R419X) results in a protein lacking 24 amino acids at its C terminus. Although this mutation has been linked to mild mental retardation, the mechanism by which the mutation affects CRBN function is unknown. Here, we used biochemical and mass spectrometric approaches to explore the function of this mutant. We show that the protein retains its ability to assemble into a Cullin-RING E3 ligase complex and catalyzes the ubiquitination of CRBN-target proteins. However, we find that this mutant exhibits markedly increased levels of autoubiquitination and is more readily degraded by the proteasome than the wild type protein. We also show that the level of the mutant protein can be restored by a treatment of cells with a clinically utilized proteasome inhibitor, suggesting that this agent may be useful for the treatment of mental retardation associated with the CRBN R419X mutation. These data demonstrate that enhanced autoubiquitination and degradation account for the defect in CRBN activity that leads to mental retardation.

Objective This study has investigated the existence of label-retaining cell and its distribution in gastric cancer, in the hope that this information will assist investigations on gastric cancer stem cells.

Peroxisome proliferator-activated receptor δ gene (PPARδ) is correlated with carcinogenesis of colorectal cancer, but the regulation of its gene transcription remains unclear. We herein report that AP1 binds the promoter and regulates PPARδ gene expression. With a luciferase reporter system, we identified a functional promoter region of 30 bp of PPARδ gene by deletion and electrophoretic mobility shift assays (EMSA). Using site-directed mutagenesis and decoy analyses, we demonstrated that AP1 bound the functional transcriptional factor binding site in a region extending from -176 to -73 of the PPARδ promoter, which was confirmed using EMSA and supershift assays. Consequently, inhibition of the AP1 binding site led to decreased PPARδ mRNA. Our study demonstrated that AP1 is the transcriptional factor that contributes to PPARδ expression in LoVo cells.

We propose and demonstrate a novel all-fiber temperature sensor that is a single-end fiber modal interferometer (FMI) based on a segment of a few mode fiber (FMF) and a segment of a single-mode fiber (SMF).The FMF spliced to a lead-in SMF is used to expend the optical beam in the fiber and the sensing SMF with a well-cleaved end is spliced to the FMF. Owing to the core mismatch between the FMF and the sensing SMF, part of the guided light propagates in the cladding of the sensing SMF and part of the guided light propagates in the core of the sensing SMF, which finally forms the FMI when the reflective light comes back to the FMF. Temperature sensing is achieved by measuring the wavelength shift of the FMI. A temperature sensor with an average sensitivity of 0.1 nm/°C in a temperature range from 33 to 833°C is presented. © 2013 Wiley Periodicals, Inc. Microwave Opt Technol Lett 55:1700–1703, 2013; View this article online at wileyonlinelibrary.com. DOI 10.1002/mop.27657

Aims We investigated the mechanisms of diabetic bladder dysfunction (BD) through analysis of the roles of L- and T-type voltage-gated calcium channels (VGCCs), with the ultimate goal of identifying potential drug targets for diabetic BD. Methods Bladder function of db/db (type 2 diabetes) and wild type (Wt) mice was evaluated by behavioral tests and in vivo cystometry. Contractile responses of bladder strips to carbachol were measured with or without pre-treatment with nifedipine (a L-type VGCC blocker) or mibefradil (a T-type VGCC blocker). Furthermore, the effects of mibefradil and nifedipine on the proliferation of human bladder smooth muscle cells (BSMCs) were studied. Results db/db mice had significantly increased voiding frequency, bladder weight, bladder compliance and capacity, and heightened contractile response to carbachol, compared to Wt mice. Nifedipine, but not mibefradil, dramatically suppressed bladder tissue contraction in Wt mice. Whereas nifedipine nearly completely inhibited bladder contraction in db/db mice, mibefradil “normalized” the heightened bladder contractility of db/db mice to the level of Wt mice. In culture, mibefradil, but not nifedipine, inhibited the proliferation of human BSMCs. Conclusion Our results indicate that while L-type VGCCs play a major role in the contraction of both diabetic and non-diabetic bladders, T-type VGCCs are involved in the contraction of diabetic bladders and mediate BSMC proliferation. This study provides support for further investigations on the effect of blockade of T-type VGCC or combined blockade of both types of VGCCs in the treatment of diabetic BD. Neurourol. Urodynam. 33:147–152, 2014. © 2013 Wiley Periodicals, Inc.

Kinases are one of the most important families of enzymes that are involved in numerous cell signaling processes. Existing methods for studying kinase expression and activation have limited kinome coverage. Herein, we established a multiple-reaction monitoring (MRM)-based targeted proteomic method that provided an unprecedented coverage (~80%) of the human kinome. We employed this method for profiling comprehensively the alterations of the global kinome of HEK293T human embryonic kidney cells upon treatment with methylglyoxal, a glycolysis byproduct that is present at elevated levels in blood and tissues of diabetic patients and is thought to contribute to diabetic complications. Our results led to the quantification of 328 unique kinases. In particular, we found that methylglyoxal treatment gave rise to altered expression of a number of kinases in the MAPK pathway and diminished expression of several receptor tyrosine kinases, including epidermal growth factor receptor (EGFR), insulin growth factor 2 receptor (IGF2R), fibroblast growth factor receptor (FGFR), etc. Furthermore, we demonstrated that the diminished expression of EGFR occurred through a mechanism that is distinct from the reduced expression of IGF2R and FGFR1. Together, our targeted kinome profiling method offers a powerful resource for exploring kinase-mediated signaling pathways that are altered by extracellular stimuli, and the results from the present study suggest new mechanisms underlying the development of diabetic complications.

The mechanisms involved in bleomycin-induced lung toxicity have not been fully understood to date. This work aimed to compare the proteome profiling of bleomycin-induced lung toxicity by using 2D-nano-LC-MS/MS and spectral counting. By comparing the spectral counts of identified proteins between control and bleomycin-treated groups, we noted that 102 proteins were upregulated and 28 proteins were downregulated in the bleomycin-treated group. Among these differently expressed proteins, five proteins were chosen for validation by Western blot analysis. The levels of these five proteins were consistent with proteomic results. These potential mediators can facilitate the translation of the underlying mechanisms of bleomycin-induced lung toxicity to molecular targets in the clinical arena.

A Disintegrin and Metalloproteinase 12 (ADAM12) is expressed significantly higher in multiple tumors than in normal tissues and has been used as a prognostic marker for the evaluation of cancer progression. Although several ADAM12 substrates have been identified biochemically and its proteolytic function has been explored, the upstream regulators and the interacting proteins have not been systematically investigated. Here, we use immunoprecipitation and mass spectrometry (MS)-based quantitative proteomic approaches to identify 28 interacting partners for the long form of ADAM12 (ADAM12-L) in HeLa cells. Proteins that regulate cell proliferation, invasion, and epithelial to mesenchymal transition are among the identified ADAM12-interacting proteins. Further biochemical experiments discover that the protein level and the stability of ADAM12 are upregulated by one of its interacting proteins, myoferlin. In addition, myoferlin also increases the proteolytic activity of ADAM12, leading to the reduction of an ADAM12 substrate, E-cadherin. This result implies that ADAM12 and its interacting proteins might converge to certain signaling pathways in the regulation of cancer cell progression. The information obtained here might be useful in the development of new strategies for modulating cell proliferation and invasion involved in the regulation between ADAM12 and its interacting partners. MS data are available via ProteomeXchange with identifier PXD003560 . Biological significance Regulation of the proliferation and invasion of cancer cells is important in cancer treatment. ADAM12 has been found to play important roles in regulating these processes and identification of its interacting partners will improve our understanding of its biological functions and provide basis for functional modulation. Through mass spectrometry-based quantitative proteomic approaches, we identify the interacting partners for ADAM12 in a human cancer cell line and find many proteins that are involved in the proliferation and invasion of cancer cells. A novel regulator, myoferlin, of ADAM12 is discovered and this protein increases ADAM12 expression level, stability, and its enzymatic activity, leading to the reduction of its substrate, E-cadherin, which plays important roles in the regulation of cell adhesion and tumor metastasis. This result provides a connection for two highly expressed proteins in cancer cells and may shed light on the regulation of their biological functions in cancer progression.

We performed this study to identify the source of complexes recorded during corpus cavernosum electromyography.A total of 10 healthy male volunteers 19 to 54 years old (mean age 36) with normal erectile function, participated in the study. A Porti electrodiagnostic system (TMS International, Enschede, The Netherlands) connected to a notebook computer recorded low frequency corpus cavernosum electromyography complexes via penile surface electrodes in 3 phases. In phase 1 baseline corpus cavernosum electromyography was recorded for 30 minutes. In phase 2 penile skin block was performed followed by 30 minutes of corpus cavernosum electromyography recording. Phase 3 consisted of intracavernous block followed by corpus cavernosum electromyography recording for 30 minutes. During all 3 phases startling auditory stimuli and median nerve stimulation were used at random intervals to evoke a sympathetic nervous system response. A suprapubic electrode was used as a control to record sympathetic electrodermal activity.In phase 1 all subjects had spontaneous and evoked corpus cavernosum complexes. The penile skin block used in phase 2 did not affect the presence of these complexes, which remained present in all subjects. During phase 3 no spontaneous or evoked corpus cavernosum complexes were recorded in any subjects, indicating that the intracavernous block eliminated the corporally generated signal. All subjects in all 3 phases demonstrated a sympathetic electrodermal response in the suprapubic electrode in response to evoking stimuli.This study confirms the penile cavernous tissue, and not the penile skin or surrounding tissue, as the origin of the corpus cavernosum electromyography complex. Further refinement of this technique may render it useful in the evaluation of neurogenic and myogenic erectile dysfunction.

Study Type – Aetiology (case control) Level of Evidence 3b What's known on the subject? and What does the study add? As one of the major components of the extracellular matrix, elastic fibres are believed to enhance tissue compliance. However, the role of elastic fibres in normal bladder function and dysfunction remains speculative. Although transgenic mice overexpressing elastin showed increased bladder compliance, the findings in patients with non-compliant bladders are inconsistent. Using transgenic elastin-deficient mouse models, this study provides the first direct evidence that sufficient elastin content is critical for healthy bladder function, and elastin is involved in the detrusor response to partial bladder outlet obstruction. OBJECTIVE • To examine functional and molecular changes of the bladders from elastin-haploinsufficient mice (Eln+/−) at baseline as well as in response to partial bladder outlet obstruction (pBOO). MATERIALS AND METHODS • Female Eln+/− and wild type (Wt) mice (3–4 months old) were studied. • The bladder elastin content was quantified by measuring desmosine. • Mice were divided into two groups to undergo surgery to create pBOO or to undergo sham surgery. Three days after surgery, bladder function was evaluated by in vivo cystometry, and the contractile response of bladder strips exposed to electrical field stimulation (EFS) and carbachol was examined by ex vivo myography. RESULTS • The Eln+/−-sham mice had a 33.6% decrease in bladder elastin compared with Wt-sham mice. • Cystometry showed significantly decreased bladder compliance and capacity in Eln+/−-sham vs Wt-sham mice; pBOO increased bladder compliance and capacity to a greater extent in Eln+/− mice compared with Wt mice. • Bladder strips from Eln+/−-sham mice showed a significantly heightened contractile response to both EFS and carbachol compared with Wt-sham mice. • A significantly increased contractile response to carbachol was detected in Wt-pBOO vs Wt-sham but not between Eln+/−-pBOO and Eln+/−-sham mice. CONCLUSION • The results that elastin-deficient mice had decreased bladder compliance and capacity and increased bladder contractility; and that Wt-pBOO mice showed an enhanced contractile response to carbachol, but Eln+/−-pBOO mice did not, suggest that elastin is critical for normal bladder function and is involved in bladder response to pBOO.

Introduction Elastin fibers confer passive recoil to many tissues including the lung, skin, and arteries. In the penis, elastin is present in sinusoids, arterioles, and in the tunica albuginea. Although decreased penile elastin has been reported in men with erectile dysfunction, the exact role of elastin in physiologic processes integral to erection remains speculative. Aim The aim of this study was to characterize erectile function in elastin-deficient mice. Methods Elastin haploinsufficient mice (Eln+/−) and aged match Eln+/+ (Wt) mice were used. Cavernosum was removed from some mice for quantification of elastin, collagen, and smooth muscle actin. Ex vivo assessment of contractile force generation was performed by myography. In vivo assessment of intracorporal pressure normalized to mean arterial pressure in response to electrical stimulation of the cavernosal nerve was measured. Veno-occlusive function was determined by cavernosography. Main Outcome Measures The main outcome measures of this study were the in vitro and in vivo assessment of cavernosal vasoreactivity, veno-occlusive function and erection in mice deficient in elastin. Results Eln +/− mice exhibited ∼33% less penile elastin than Wt mice, with no change in collagen. Cavernosal tissue from Eln+/− mice has a significantly heightened contractile response, explained in part by increased smooth muscle cell content. Veno-occlusive function was significantly altered in Eln+/− mice. Interestingly, erectile function was impaired only at submaximal voltage (1 V) stimulation (there was no impairment during the higher 2-V stimulus). Conclusion Eln +/− mice display a cavernosal phenotype consistent with developmental changes attributable to the loss of elastin. These alterations confer a degree of altered erectile function that is able to be overridden by maximal stimulatory input. Altogether, these data suggest that elastin is important for erectile function.

Nitric oxide (NO) is considered as a hallmark for allergic airway inflammation in asthmatics and animal models. But the correlation between NO and antigen-induced pulmonary inflammation (AIPI), a rat model for asthma, in varying genetic background population has not been completely understood.The objective in this study is to observe the different responsiveness to AIPI in two commonly used inbred rat strains and verify the correlation between NO from different sources and pathological parameters of AIPI by using Dark Agouti (DA), E3, F1 (E3 x DA), and F2 rat populations.AIPI was induced by systemically immunizing and intranasally challenging E3, DA, F1 (DA x E3), and F2 rats with ovalbumin (OVA). Pathological changes and mucus secretion in lungs were observed after hematoxylin and eosin (HE) and periodic acid Schiff (PAS) staining, whereas eosinophils in bronchoalveolar lavage fluid (BALF) were counted after Giemsa staining. Delayed-type hyperresponsiveness was determined by subcutaneous injection of OVA in ear. Total immunoglobulin E (IgE) and OVA-specific IgG1 were detected with enzyme-linked immunosorbent assay (ELISA). NO concentration was measured by the Griess method.DA rats were unresponsive to OVA treatment, whereas E3 rats were susceptible to AIPI. F1 rats manifested the same responsiveness to OVA treatment as DA rats, and individual F2 rats showed the variable severity of AIPI. NO concentration in BALF and serum was significantly elevated in E3 rats but not in DA and F1 rats after OVA treatment. In F2 rats, NO concentration in serum was positively correlated with eosinophils in BALF, total IgE, and pathological scores, whereas NO concentration in BALF correlated only with eosinophils in BALF and total IgE.DA and F1 rats are resistant, whereas E3 rats are sensitive, to AIPI. NO in serum can represent the severity of allergic inflammation and pathological changes in lungs in F2 population.

Objective Retrospective assessment of short and long-term outcome in elderly undergoing primary isolated Valvular Heart Surgery (VR).

Although great progresses have been made in proteomics during the last decade, proteomics is still in its infancy. Extreme complexity of proteome sample and large dynamic range of protein abundance overwhelm the capability of all currently available analytical platforms. Sample pretreatment is a good approach to reduce the complexity of proteome sample and decrease the dynamic range. In this article, we present an overview of different technologies and methods for sample pretreatment in efficient proteome and peptidome analysis. Methods for isolation of rare amino acid-containing peptides, terminal peptides, PTM peptides and endogenous peptides are reviewed. In addition, two automated sample pretreatment technologies, i.e. automated sample injection and on-line digestion, are also covered.