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Butyrophilin 3A1 (BTN3A1), a major histocompatibility complex‐associated gene that encodes a membrane protein with two extracellular immunoglobulin domains and an intracellular B30.2 domain, is critical in T‐cell activation and adaptive immune response. Here, the expression of BTN3A1 in cancers was analyzed in eight databases comprising 86 733 patients of 33 cancers, and the findings were validated in patient samples and cell models. We showed that BTN3A1 was expressed in most cancers, and its expression level was strongly correlated with clinical outcome of 13 cancers. Mutations of BTN3A1 were detected, and the mutations were distributed throughout the entire gene. Gene set enrichment analysis showed that BTN3A1 co‐expression genes and interacting proteins were enriched in immune regulation‐related pathways. BTN3A1 was associated with tumor‐infiltrating immune cells and was co‐expressed with multiple immune checkpoints in patients with breast cancer (BRCA) and non‐small cell lung cancer (NSCLC). We reported that BTN3A1 was downregulated in 46 of 65 (70.8%) NSCLCs, and its expression level was inversely associated with clinical outcome of the patients. BTN3A1 in tumor samples was lower than in counterpart normal tissues in 31 of 38 (81.6%) BRCAs. Bioinformatics analyses showed that BTN3A1 could be a target gene of transcription factor Spi‐1 proto‐oncogene (SPI1), and our ‘wet’ experiments showed that ectopic expression of SPI1 upregulated, whereas silencing of SPI1 downregulated, BTN3A1 expression in cells. These results suggest that BTN3A1 may function as a tumor suppressor and may serve as a potential prognostic biomarker in NSCLCs and BRCAs., BTN3A1 was altered in most cancers and was downregulated and positively associated with prognosis of patients with non‐small cell lung cancer and breast cancer. BTN3A1 was correlated with immune cell infiltration and immune checkpoints' expression and was a target gene of transcription factor SPI1. It may function as a tumor suppressor and plays a role in carcinogenesis.

Methylobacterium extorquens AM1 (AM1), a model strain of methylotrophic cell factories (MeCFs) could be used to produce fine chemicals from methanol. Synthesis of heterologous products usually needs reducing cofactors, but AM1 growing on methanol lack reducing power. Formate could be used as a reducing agent. In this study, mevalonic acid (MEV) yield of 0.067 gMEV/g methanol was reached by adding 10 mmol L

Integrin alpha x (ITGAX), a member of the integrin family, usually serves as a receptor of the extracellular matrix. Recently, accumulating evidence suggests that ITGAX may be involved in angiogenesis in dendritic cells. Herein, we report a direct role of ITGAX in angiogenesis during tumor development. Overexpression of ITGAX in human umbilical vein endothelial cells (HUVECs) enhanced their proliferation, migration, and tube formation and promoted xenograft ovarian tumor angiogenesis and growth. Further study showed that overexpression of ITGAX activated the PI3k/Akt pathway, leading to the enhanced expression of c-Myc, vascular endothelial growth factor-A (VEGF-A), and VEGF receptor 2 (VEGFR2), whereas, the treatment of cells with PI3K inhibitor diminished these effects. Besides, c-Myc was observed to bind to the VEGF-A promoter. By Co-Immunoprecipitation (Co-IP) assay, we manifested the interaction between ITGAX and VEGFR2 or the phosphorylated VEGFR2. Immunostaining of human ovarian cancer specimens suggested that endothelial cells of micro-blood vessels displayed strong expression of VEGF-A, c-Myc, VEGFR2, and the PI3K signaling molecules. Also, overexpression of ITGAX in HUVECs could stimulate the spheroid formation of ovarian cancer cells. Our study uncovered that ITGAX stimulates angiogenesis through the PI3K/Akt signaling-mediated VEGFR2/VEGF-A overexpression during cancer development.

The circadian clock enables organisms to rapidly adapt to the ever-changing environmental conditions that are caused by daily light/dark cycles. Circadian clock genes universally affect key agricultural traits, particularly flowering time. Here, we show that OsPRR37, a circadian clock gene, delays rice flowering time in an expression level-dependent manner. Using high-throughput mRNA sequencing on an OsPRR37 overexpressing transgenic line (OsPRR37-OE5) and the recipient parent Guangluai4 that contains the loss-of-function Osprr37, we identify 14,992 genes that display diurnal rhythms, which account for 52.9% of the transcriptome. Overexpressing OsPRR37 weakens the transcriptomic rhythms and alters the phases of rhythmic genes. In total, 3,210 differentially expressed genes (DEGs) are identified, among which 1,863 rhythmic DEGs show a correlation between the change of absolute amplitudes and the mean expression levels. We further reveal that OsPRR37 functions as a transcriptional repressor to repress the expression levels and amplitudes of day-phased clock genes. More importantly, OsPRR37 confers expanded regulation on the evening-phased rhythmic DEGs by repressing the morning-phased rhythmic DEGs. Further study shows that OsPRR37 expands its regulation on flowering pathways by repressing Ehd1. Thus, our results demonstrate an expanded regulation mechanism of the circadian clock on the diurnal rhythms of the transcriptome.

Fine particulate matter (PM2.5 ) is a major component of air pollutions that are closely associated with increased risk of lung cancer. However, the role of PM2.5 in the etiology of lung cancer is largely unknown. In this study, we performed acute (24 hours) and chronic (five passages) exposure models to investigate the carcinogenetic mechanisms of PM2.5 by targeting the induction of epithelial-mesenchymal transition (EMT) and cancer stem cells (CSC) properties in human non-small cell lung cancer cell line A549. We found that both acute and chronic PM2.5 exposure enhanced cell migration and invasion, decreased mRNA expression of epithelial markers and increased mRNA expression of mesenchymal markers. Chronic PM2.5 exposure further induced notable EMT morphology and CSC properties, indicating the developing process of cell malignant behaviors from acute to chronic PM2.5 exposure. CSC properties induced by chronic PM2.5 exposure characterized with increased cell-surface markers (CD44, ABCG2), self-renewal genes (SOX2 and OCT4), side population cells and neoplastic capacity. Furthermore, the levels of three stemness-associated microRNAs, Let-7a, miR-16 and miR-34a, were found to be significantly downregulated by chronic PM2.5 exposure, with microarray data analysis from TCGA database showing their lower expression in human lung adenocarcinoma tissues than that in the adjacent normal lung tissues. These data revealed that the induction of EMT and CSC properties were involved in the lung cancer risk of PM2.5 , and implicated CSC properties and related microRNAs as possible biomarkers for carcinogenicity prediction of PM2.5 .

Reactive flash volatilisation is an autothermal process to convert biomass into tar-free synthesis gas under steam-rich conditions. This article studies the kinetics of reactive flash volatilisation by using Ni, PtNi, RuNi, ReNi, and RhNi catalysts supported on alumina. The rates of mass loss of cellulose, xylan, and lignin were measured and compared with those of the synthetic biomass mixture and pinewood sawdust. The kinetic parameters were calculated with and without catalysts by using a wire-mesh isothermal thermogravimetric analyser in an equimolar steam/N2 atmosphere and high heating rates of 8.6×102, 1.1×103, and 1.3×104 °C min−1 at 700, 750, and 800 °C, respectively. The results revealed three distinct regimes of the rate of mass loss: pyrolytic decomposition, reforming, and char gasification. The catalysts increased the rate of mass loss in the reforming regime. RhNi and RuNi supported catalysts showed higher reforming rates than other catalysts. This study provides direct evidence of the in situ catalytic removal of tar during gasification of biomass.

Biomass gasification is considered to be one of the most promising technologies to deliver renewable energy. However, tar formation in the gasifier is one of the main challenges. Ni-based catalysts are one of the most effective transition-metal catalysts in biomass gasification for tar cracking and reforming. Alumina-supported Ni, Pt-Ni, Ru-Ni, Re-Ni and Rh-Ni catalysts were tested for their activity in the reactive flash volatilisation (RFV) of cellulose to produce synthesis gas in 50 ms reaction time. RFV is a catalytic gasification process that utilises a high carbon space velocity and mass flow rate with oxygen and steam as gasification agents. Re-Ni, Rh-Ni and Ru-Ni supported catalysts showed a higher gasification efficiency than the other catalysts, which was because of their higher metal surface area, high reducibility and lower CO desorption temperature. The highest gasification efficiency was achieved at 750 °C with a carbon-to-oxygen ratio of 0.6 and a carbon-to-steam ratio of 1.0 without any oxygen breakthrough.

Background The development of genetically modified pigs, which lack the expression of alpha 1-3 galactosyl transferase, (GalT-KO pigs) has facilitated the xenogeneic transplantation of porcine organs and tissues into primates by avoiding hyperacute rejection due to pre-existing antibodies against the Gal epitope. However, antibodies against other antigens (anti-non-Gal antibodies), are found at varying levels in the pre-transplant sera of most primates. We have previously found that baboons with high levels of pre-transplant anti-non-Gal IgG, conditioned with a non-myeloablative conditioning regimen, failed to engraft following pig-to-baboon bone marrow transplantation (Xenotransplantation, 17, 2010 and 300). Two baboons with low levels of pre-transplant anti-non-Gal IgG, conditioned with the same regimen, showed porcine bone marrow progenitors at 28 days following transplantation, suggesting engraftment. These baboons also showed evidence of donor-specific hyporesponsiveness. This observation led us to investigate the hypothesis that selecting for baboon recipients with low pre-transplant anti-non-Gal IgG levels might improve engraftment levels following GalT-KO pig-to-baboon bone marrow transplantation. Methods Five baboons, with low pre-transplant anti-non-Gal IgG levels, received transplantation of bone marrow cells (1–5 × 109/kg of recipient weight) from GalT-KO pigs. They received a non-myeloablative conditioning regimen consisting of low-dose total body irradiation (TBI) (150 cGy), thymic irradiation (700 cGy), anti-thymocyte globulin (ATG), and tacrolimus. In addition, two baboons received Rituximab and Bortezomib (Velcade) treatment as well as extra-corporeal immunoadsorption using GalT-KO pig livers. Bone marrow engraftment was assessed by porcine-specific PCR on colony forming units (CFU) of day 28 bone marrow aspirates. Anti-non-Gal antibody levels were assessed by serum binding toward GalT-KO PBMC using flow cytometry (FACS). Peripheral macro-chimerism was measured by FACS using pig and baboon-specific antibodies and baboon anti-pig cellular responses were assessed by mixed lymphocyte reactions (MLR). Results As previously reported, two of five baboons demonstrated detectable bone marrow engraftment at 4 weeks after transplantation. Engraftment was associated with lack of an increase in anti-non-Gal IgG levels as well as cellular hyporesponsiveness toward pig. Three subsequent baboons with similarly low levels of pre-existing anti-non-Gal IgG showed no engraftment and an increase in anti-non-Gal IgG antibody levels following transplantation. Peripheral macrochimerism was only seen for a few days following transplantation regardless of antibody development. Conclusions Selecting for baboon recipients with low levels of pre-transplant anti-non-Gal IgG did not ensure bone marrow engraftment. Failure to engraft was associated with an increase in anti-non-Gal IgG levels following transplantation. These results suggest that anti-non-Gal-IgG is likely involved in early bone marrow rejection and that successful strategies for combating anti-non-Gal IgG development may allow better engraftment. Since engraftment was only low and transient regardless of antibody development, innate immune, or species compatibility mechanisms will likely also need to be addressed to achieve long term engraftment.

Natural products have proven to be valuable sources for the identification of new drug candidates, and also as tools for chemical biology and medicinal chemistry research [1]. In fact, if we trace back human history, many natural products have also long been used to treat various human disorders and distinguished by their drug-like properties [2]. To date, tens of thousands of bioactive compounds have been isolated from plants, microbes, marine invertebrates, and other sources [3]. Consequently, these chemical structures have been employed by chemists as references to scan the diversity space for drug discovery efforts [4]. It is estimated that 50–70% of launched drugs in the marketplace are either natural products themselves or natural product-derived molecules [5]. So, what features of natural products make them effective drug candidates? Natural products play important roles in biomedical research and drug discovery largely attributable to their structural complexity and diversity, which Nature has engineered to facilitate optimal functions of living systems. Structural complexity and diversity enable natural products to modulate biological targets of human diseases. The drug-likeness of natural products often possesses common factors including molecular complexity, ability to bind to proteins, structural rigidity, and three-dimensionality [6]. Comparedwith synthetic molecules, the chemical structures of natural products are more constrained, which allows the accumulation of reliable structure–activity relationship (SAR) data for studying protein target–chemical ligand interactions [7]. Recently, much attention has been devoted to the natural products which are an obvious violation of ‘‘Rule-of-Five’’ [8], but still possess reasonable biopharmaceutical and pharmacokinetic properties, for example, ciclosporin A. Thus, exploitation of such types of natural products could not only significantly broaden the chemical space beyond the ‘‘Rule-of-Five’’ domain for the purpose of drug discovery, but also give us the opportunity to appreciate and investigate how Nature selects and optimizes natural products which are able to bind to and disrupt the function of biological targets through evolution. We should also recognize that natural products were optimized in living systems presumably not for the same purpose as is desired to serve in a biomedical research

Griesemer A, Liang F, Hirakata A, Hirsh E, Lo D, Okumi M, Sykes M, Yamada K, Huang CA, Sachs DH. Occurrence of specific humoral non-responsiveness to swine antigens following administration of GalT-KO bone marrow to baboons. Xenotransplantation 2010; 17: 300–312. © 2010 John Wiley & Sons A/S. Abstract: Background: Hematopoietic chimerism induces transplantation tolerance across allogeneic and xenogeneic barriers, but has been difficult to achieve in the pig-to-primate model. We have now utilized swine with knockout of the gene coding for α-1,3-galactosyltransferase (GalT-KO pigs) as bone marrow donors in an attempt to achieve chimerism and tolerance by avoiding the effects of natural antibodies to Gal determinants on pig hematopoietic cells. Methods: Baboons (n = 4; Baboons 1 to 4 = B156, B158, B167, and B175, respectively) were splenectomized and conditioned with TBI (150 cGy), thymic irradiation (700 cGy), T cell depletion with rabbit anti-thymocyte globulin (rATG) and rat anti-primate CD2 (LoCD2b), and received FK506 and supportive therapy for 28 days. All animals received GalT-KO bone marrow (1 to 2 × 109 cells/kg) in two fractions on days 0 and 2, and were thereafter monitored for the presence of pig cells by flow cytometry, for porcine progenitor cells by PCR of BM colony-forming units, and for cellular reactivity to pig cells by mixed lymphocyte reaction (MLR). In vitro antibody formation to LoCD2b and rATG was tested by ELISA; antibody reactivity to GalT-KO pig cells was tested by flow cytometry and cytotoxicity assays. Additionally, Baboons 3 and 4 received orthotopic kidney transplants on days 17 and 2, respectively, to test the potential impact of the protocol on renal transplantation. Results: None of the animals showed detectable pig cells by flow cytometry for more than 12 h post-BM infusion. However, porcine progenitor cell engraftment, as evidenced by pig-derived colony forming units in the BM, as well as peripheral microchimerism in the thymus, lymph node, and peripheral blood was detected by PCR in baboons 1 and 2 for at least 28 days post-transplant. ELISA results confirmed humoral immunocompetence at time of transplantation as antibody titers to rat (LoCD2b) and rabbit (ATG) increased within 2 weeks. However, no induced antibodies to GalT-KO pig cells or increased donor specific cytotoxicity was detectable by flow cytometry. In contrast, baboons 3 and 4 developed serum antibodies to pig cells as well as to rat and rabbit immunoglobulin by day 14. Retrospective analysis revealed that although all four baboons possessed low levels of antibody-mediated cytotoxicity to GalT-KO cells prior to transplantation, the two baboons (3 and 4) that became sensitized to pig cells (and rejected pig kidneys) had relatively high pre-transplantation titers of anti–non-Gal IgG detectable by flow cytometry, whereas baboons 1 and 2 had undetectable titers. Conclusions: Engraftment and specific non-responsiveness to pig cells has been achieved in two of four baboons following GalT-KO pig-to-baboon BMT. Engraftment correlated with absence of preformed anti–non-Gal IgG serum antibodies. These results are encouraging with regard to the possibility of achieving transplantation tolerance across this xenogeneic barrier.

Methylobacterium extorquens AM1, which can be used as a methylotrophic cell factory (MeCF) for the production of fine chemicals from methanol, is the most extensively studied model methylotrophic strain. However, its low tolerance for methanol limits the development of bioprocesses and there have been no reports of improved methanol tolerance of M. extorquens AM1. In this study, atmospheric and room temperature plasma (ARTP) mutagenesis, in combination with adaptive laboratory evolution (ALE), is used to generate a mutant with high methanol tolerance (referred to as CLY-2533). The final cell density of CLY-2533 is 7.10 times higher than that of the wild-type strain in medium containing 5% (v/v) methanol. Through comparative genomics analysis and overexpression of the exploited putative genes, seven mutated genes are identified as being closely related to the higher methanol tolerance of CLY-2533. Additionally, the mvt operon, which contains genes related to the biosynthesis of mevalonate acid (MEV), is introduced into CLY-2533. This recombinant strain shows significant improvements in both MEV production and cell growth in 5% methanol medium. These findings will be helpful in rational design of methanol-utilizing strain for an improved host platform for methanol based biomanufacturing.

* These two authors contribute equally to this paper. In order to identify the proteins associated with Helicobacter pylori colonization in mice, we used 2-dimensional gel electrophoresis (2-DE) to analyze the membrane- and soluble-cellular proteins extracted from H. pylori strain 26695 and the mouse-passaged homolog 88-3887. We defined 2- and 3-fold changes in protein expression as the threshold values for differential expression in the membrane-protein and whole-cell-protein fractions, respectively. The differentially expressed proteins were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/TOF). A total of 29 proteins, including 16 mem- brane- or membrane-associated proteins (13 upregulated, 3 downregulated) and 13 cellular proteins (10 upregulated, 3 downregulated) were differentially expressed between the strains 26695 and 88-3887. Among the upregulated proteins, 10 proteins had been previously shown to be associated with the mouse colonization, and 13 upregulated proteins were shown to be associated with the adaptation of H. pylori in murine hosts for the first time in this study. The identified proteins were classified as proteins related to metabolism, stress response, virulence, or adhesion. The data presented in this report indicated that there were subsets of upregulated proteins in mouse-adapted H. pylori. In particular, the adhesins, virulence factors, and stress- response proteins are likely to contribute to colonization in mice.

The correlation between morphology and charge-carrier mobility in the vertical direction in thin films of poly(2-methoxy-5-(2′ethylhexyloxy)-1,4-phenylenevinylene) (MEH-PPV) is investigated by a combination of X-ray reflectivity (XRR), field-emission scanning electron microscopy (FESEM), atomic force microscopy (AFM), fluorescence optical microscopy (FOM), photoluminescence spectroscopy (PL), photoluminescence excitation spectroscopy (PLE), as well as time-of-flight (TOF) and transient electroluminescence (TrEL) techniques. The mobility is about two orders of magnitude greater for drop-cast films than for their spin-cast counterparts. Drop-casting in the presence of a vertical static electric field (E-casting) results in films with an additional increase in mobility of about one order of magnitude. While PL and PLE spectra vary with the method of film preparation, there is no correlation between emission spectra and charge-carrier mobility. Our XRR measurements on spin-cast films indicate layering along the film depth while no such structure is found in drop-cast or E-cast films, whereas FESEM examination indicates that nanodomains within drop-cast films are eliminated in the E-cast case. These observations indicate that carrier transport is influenced by structure on two different length scales. The low mobility observed in spin-cast films is a direct result of a global layered structure with characteristic thickness of ca. 4 nm: in the absence of this layered structure, drop-cast films with inherent nanoscale heterogeneities (ca. 20 nm in size) exhibit much better hole mobility. Elimination of nanodomains via electric-field alignment results in further improved charge mobility.

Different thermal processing methods were used to fabricate the crystalline properties of poly(vinylidene fluoride-co-chlorotrifluoroethylene) [P(VDF-co-CTFE)] films. We observed that the crystallinity and crystal grain size of the various samples decreased with the quenching temperature. Compared to that of the annealed P(VDF-co-CTFE) sample, a higher dielectric constant of 13.9 at a frequency of 100 Hz was obtained in the film with liquid nitrogen quenching because the increasing small crystalline regions were susceptible to the excitation of external electric field. Meanwhile, the breakdown electric strength of the low-temperature-quenched film increased to 530 MV/m when the depth of shallow electronic energy level decreased, as depicted by Frohlich collective electron approximated electric breakdown theory. Moreover, when we introduced the leakage current density curves, the effect of the space charges on the electric displacement was proven. As a result, the discharged energy density of the liquid-nitrogen-quenched P(VDF-co-CTFE) film was enhanced to 15.32 J/cm3 at an electric field of 530 MV/m; this provided an effective way in addition to chemical modification to achieve a high energy storage ability in this poly(vinylidene fluoride)-based fluoropolymer. © 2015 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2015, 132, 42794.

A nanostructure surface treatment is proposed to improve the response time and driving voltage of cholesteric liquid crystal (CLC) devices. With nanostructure surface, threshold Voltage (Vth) can be reduced ∼10% with minimum reflectance loss. The transition time can be greatly reduced by 80% and multi-level gray scale can be achieved with new driving scheme in the nanostructure enhanced CLC cell.