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Glyphosate-resistant invasive plants, including Amaranthus palmeri S. Watson, have greatly challenged management of new invasions. Elucidating their glyphosate resistance levels rapidly and accurately will better inform management strategies. Quantitative real-time PCR (qPCR) has been used to identify glyphosate resistance in A. palmeri by detecting gene copy numbers of 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS), an enzyme inhibited by glyphosate. However, qPCR can only indirectly determine copy numbers because it requires a calibrator sample; it also lacks standardization, thus limiting its usefulness. Droplet digital PCR (ddPCR) is a new method to detect copy number directly and precisely. We evaluated ddPCR as a tool to determine glyphosate-resistance level while using qPCR and glyphosate dose response (GDR) assays as reference technologies to compare performance and efficiency between methods.We identified seven susceptible and seven resistant populations of A. palmeri using the GDR assay. Resistant levels detected by qPCR and ddPCR were generally consistent with the GDR results. Although detected values obtained by qPCR and ddPCR were highly correlated (ROur findings demonstrate that ddPCR offers improved accuracy and specificity in detecting EPSPS gene copy numbers and is a robust and rapid method for glyphosate-resistance identification in A. palmeri. Our research is the first to measure glyphosate resistance in A. palmeri by ddPCR assay and will shed light on future applications of ddPCR in identifying herbicide resistance in other invasive weeds. © 2022 Society of Chemical Industry.

Oral squamous cell carcinoma (OSCC) has a very poor prognosis because of its highly invasive nature, and the 5-year survival rate has not changed appreciably for the past 30 years. Although cylindromatosis (CYLD), a deubiquitinating enzyme, is thought to be a potent tumour suppressor, its biological and clinical significance in OSCC is largely unknown. This study aimed to clarify the roles of CYLD in OSCC progression. Our immunohistochemical analyses revealed significantly reduced CYLD expression in invasive areas in OSCC tissues, whereas CYLD expression was conserved in normal epithelium and carcinoma in situ. Furthermore, downregulation of CYLD by siRNA led to the acquisition of mesenchymal features and increased migratory and invasive properties in OSCC cells and HaCaT keratinocytes. It is interesting that CYLD knockdown promoted transforming growth factor-β (TGF-β) signalling by inducing stabilization of TGF-β receptor I (ALK5) in a cell autonomous fashion. In addition, the response to exogenous TGF-β stimulation was enhanced by CYLD downregulation. The invasive phenotypes induced by CYLD knockdown were completely blocked by an ALK5 inhibitor. In addition, lower expression of CYLD was significantly associated with the clinical features of deep invasion and poor overall survival, and also with increased phosphorylation of Smad3, which is an indicator of activation of TGF-β signalling in invasive OSCC. These findings suggest that downregulation of CYLD promotes invasion with mesenchymal transition via ALK5 stabilization in OSCC cells. Copyright © 2017 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.

The alternative development of coal-bearing hydrocarbon source rocks and low-porosity and low-permeability tight sandstone reservoirs of the Triassic Xujiahe Formation in the Sichuan Basin is favorable for near-source hydrocarbon accumulation. The natural gas composition of the Xujiahe Formation in the Sichuan Basin is dominated by hydrocarbon gases, of which the methane content is 80.16%–98.67%. Typically, the C2+ content is larger than 5% in main wet gas. The dry gas is mainly distributed in the western and northern regions of the basin. The non-hydrocarbon gases mainly contain nitrogen, carbon dioxide, hydrogen, and helium, with a total content of δ13C4 occurs in some natural gas with a low evolution degree; such gas is mainly coal-related gas from humic-type source rocks of the Xujiahe Formation. As for the natural gas, δ2HCH4 values ranged from −195‰ to −161‰, δ2HC2H6 values ranged from −154‰ to −120‰, and δ2HC3H8 values ranged from −151‰ to −108‰. The dry coefficient, δ13C and δ2HCH4 are all positively correlated with the maturity of source rocks. The higher the maturity of source rocks is, the larger the natural gas dry coefficient is and the larger the δ13C and δ2HCH4 values are, indicative of the characteristic of near-source accumulation. The δ2HC2H6 value of natural gas is influenced by paleosalinity to a relatively large extent; the higher the paleosalinity is, the larger the δ2HC2H6 value is. The Pr/Ph value of the condensate oil ranged from 1.60 to 3.43, illustrating light oxidization–light reduction and partial-oxidization characteristics of the depositional environment of coal-bearing source rocks of the Xujiahe Formation. The natural gas light hydrocarbon (C5–C7) from the Xujiahe Formation presented two characteristics: the first was the relatively high aromatic hydrocarbon content (19%–32.1%), which reveals the characteristic of natural gas with humic substances of high-maturity; the second was the low content of aromatic hydrocarbon (0.4%–9.3%), reflecting water-washing during the accumulation of the natural gas. The reported research outcomes indicate a potential mechanism for natural gas accumulation in the Xujiahe Formation, which will further guide natural gas exploration in this region.

Cancer stem cells (CSCs) drive tumor initiation and metastasis in several types of human cancer. However, the contribution of ovarian CSCs to peritoneal metastasis remains unresolved. The cell adhesion molecule CD44 has been identified as a major marker for CSCs in solid tumors, including epithelial ovarian cancer. CD44 exists as a standard form (CD44s) and also as numerous variant isoforms (CD44v) generated by alternative mRNA splicing. Here we show that disseminated ovarian tumors in the pelvic peritoneum contain highly enriched CD44v6-positive cancer cells, which drive tumor metastasis and are responsible for tumor resistance to chemotherapy. Clinically, an increased number of CD44v6-positive cancer cells in primary tumors was associated with a shortened overall survival in stage III-IV ovarian cancer patients. Furthermore, a subpopulation of CD44v6-positive cancer cells manifested the ability to initiate tumor metastasis in the pelvic peritoneum in an in vivo mouse model, suggesting that CD44v6-positive cells show the potential to serve as metastasis-initiating cells. Thus, the peritoneal disseminated metastasis of epithelial ovarian cancer is initiated by the CD44v6-positive subpopulation, and CD44v6 expression is a biomarker for the clinical outcome of advanced ovarian cancer patients. Given that a distinct subpopulation of CD44v6-positive cancer cells plays a critical role in peritoneal metastasis, definitive treatment should target this subpopulation of CD44v6-positive cells in epithelial ovarian cancer.

Aims Neutrophil gelatinase-associated lipocalin (NGAL) is a member of the lipocalin superfamily. Although its overexpression in various cancers has been reported, little is known about its expression and clinical significance in oral squamous cell carcinoma (OSCC). This study aimed to elucidate the clinical significance of NGAL in OSCC. Methods and results We investigated NGAL expression immunohistochemically in tumour cells and stromal cells in 96 OSCC tissues. NGAL expression in tumour cells correlated significantly with histological tumour cell differentiation, as shown by its specific distribution in the horn pearl-forming keratinized tumour cells, but not with other major clinicopathological parameters. We found NGAL+ cells in the stroma that were predominantly myeloperoxidase-positive neutrophils. The number of such NGAL-expressing stromal cells was associated significantly with poor differentiation and reduced overall survival in OSCC. The prognostic value of stromal NGAL expression was significant in a univariate analysis, while only a trend was found in multivariate analyses. Conclusions This study is the first to show the clinical significance of stromal NGAL expression, which may be an indicator of poor prognosis and more aggressive histological grade in OSCC. Our data suggest that NGAL expression in tumour cells and expression in stroma are associated in different ways with OSCC differentiation.

Abies fabri (Mast.) Craib. (A. fabri) is an endemic and dominant species in typical subalpine dark coniferous forests distributed in mountainous regions of Western Sichuan, China. We investigated the ecophysiological responses of A. fabri seedlings to short-term experimental drought, nitrogen supply and their interaction. Drought stress was created by excluding natural precipitation with automatically controlled plastic roof that covered the seedlings. Nitrogen fertilization was applied weekly by spraying over seedlings ammonium nitrate solution (50 kg N ha(-1) year(-1)) during the growing season of 2009. The results showed that drought stress decreased leaf relative water content (RWC), whereas it caused marked increases in root mass ratio (RMR) and root/shoot mass ratio by 6.19 and 10.39%, respectively, as compared with the control. Drought stress increased malondialdehyde (MDA) content, electrolyte leakage, proline content, soluble sugars content and the activities of antioxidant enzymes, whereas nitrogen supply decreased MDA content, but enhanced activities of some antioxidant enzymes [especially peroxidase (POD)]. In the drought stressed plots, nitrogen supply increased RWC and decreased the content of MDA. The combination of drought stress and nitrogen supply also decreased the activities of antioxidant enzymes. These results indicated that the negative effects of drought stress on A. fabri seedlings might be alleviated by nitrogen supply.