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Accurate detection of underwater objects is a key indicator technology to effectively enhance the field of marine development and application, and is of great importance to various fields including marine military defense and seafood aquaculture. Efficient and rapid detection of underwater targets is a crucial technological challenge in this field. To meet the challenges posed by these issues, this study applies the convolutional omni‐efficient layer aggregation network (CO‐ELAN) module to the detector backbone to improve the ability of the network structure to acquire underwater objects from image information. The module improves the feature representation of gradient branching through a multi‐dimensional dynamic convolution and attention mechanism. In terms of loss calculation, the optimized normalized Wasserstein distance approach is used to predict the box distribution probabilistic modelling method to determine comparable distances to the ground box and obtain better samples of small target labels. Here, an underwater image enhancement algorithm based on white balance and underwater blur fusion is used to obtain clear images that enable improved detector performance. After the verification experiment on the URPC2018 dataset, it is found that the detector has better underwater detection ability compared with other detectors in the complex underwater environment. The proposed method achieves a 2.4% improvement over the YOLOv7 baseline model, while reducing computation costs by 5%. [ABSTRACT FROM AUTHOR]

Dissimilar components composed of CFRTP (Carbon Fiber Reinforced Thermoplastic Polymer) and TC4 titanium alloy are increasingly applied in the aerospace field. The scanning mode offers a significant influence on the quality of laser joining joint between CFRTP and TC4 titanium alloy. Therefore, the laser joining between TC4 titanium alloy with surface microgrooves and CFRTP has been implemented under oscillating laser joining mode and linear laser joining mode respectively in the present research. The temperature distribution is qualitatively explored based on the established mathematical model of laser joining between CFRTP and TC4 titanium alloy. The interface morphology and the joining strength of CFRTP/TC4 titanium alloy lap joints under oscillating laser joining and linear laser joining are compared. The results indicate that the simulated temperature distribution shows good agreement with the experimental results. Compared with linear laser joining, the oscillating laser joining weakens the heat concentration and creates a heating zone with larger area and more uniform temperature distribution. The interface morphology of laser joining CFRTP/TC4 titanium alloy joints obtained by oscillating laser joining presents better resin filling and fewer bubble defects due to the temperature variation of the form of unequal amplitude oscillations, with the resin filling ratio of 92.20% and the porosity of 3.78%. In contrast, the linear laser scanning mode with a large number of large-sized bubbles in the filling resin and small-sized fusion gaps distributed at the interface holds a resin filling rate of 60.11% and a porosity of 32.89%. By adopting the joining method with oscillating laser scanning mode, higher quality joints can be obtained with the joining strength of 24.48 MPa. [ABSTRACT FROM AUTHOR]

[Display omitted] • Imidacloprid-resistant N. lugens population rapidly adapted to resistant rice IR36. • Insecticide resistance-related genes nAChR - 7 - like and CYP4C61 contributed to the adaptation to resistant rice in N. lugens. • One avirulent/susceptible genotype and two virulent/resistant genotypes could be inferred from the corresponding alleles of genes nAChR - 7 - like and CYP4C61. • The virulent/resistant genotypes of N. lugens already existed in the wild in China with increasing frequencies along with insecticide usage. Preventing crop yield loss caused by pests is critical for global agricultural production. Agricultural pest control has largely relied on chemical pesticides. The interaction between insecticide resistance and the adaptation of herbivorous pests to host plants may represent an emerging threat to future food security. This study aims to unveil genetic evidence for the reduction in the profitability of resistant cultivars derived from insecticide resistance in target pest insects. An experimental evolution system encompassing resistant rice and its major monophagous pest, the brown planthopper Nilaparvata lugens , was constructed. Whole genome resequencing and selective sweep analysis were utilized to identify the candidate gene loci related to the adaptation. RNA interference and induced expression assay were conducted to validate the function of the candidate loci. We found that the imidacloprid-resistant population of N. lugens rapidly adapted to resistant rice IR36. Gene loci related to imidacloprid resistance may contribute to this phenomenon. Multiple alleles in the nicotinic acetylcholine receptor (nAChR)- 7 - like and P450 CYP4C61 were significantly correlated with changes in virulence to IR36 rice and insecticide resistance of N. lugens. One avirulent/susceptible genotype and two virulent/resistant genotypes could be inferred from the corresponding alleles. Importantly, we found that the virulent/resistant genotypes already exist in the wild in China, exhibiting increasing frequencies along with insecticide usage. We validated the relevance of these genotypes and the virulence to three more resistant rice cultivars. Knockdown of the above two genes in N. lugens significantly decreased both the resistance to imidacloprid and the virulence towards resistant rice. Our findings provide direct genetic evidence to the eco-evolutionary consequence of insecticide resistance, and suggest an urgent need for the implementation of predictably sustainable pest management. [ABSTRACT FROM AUTHOR]

Ulcerative colitis (UC) is believed to arise from an imbalance between the intestinal microbiota and mucosal immunity, leading to excessive intestinal inflammation. Modulating the gut microbial community through dietary components presents a valuable strategy in aiding the treatment of UC. In this study, esters formed by binding of well-known prebiotics, fructooligosaccharides (FOS), with short chain fatty acids (SCFAs) via both enzymatic and chemical methods were evaluated for their impact on the gut microbiota of UC patients. An in vitro human colonic fermentation model was employed to monitor changes in total carbohydrates and SCFAs production during the fermentation of these esters by microbiota from patients with active and remission UC. The results showed that pronounced abundance of [Ruminococcus]_gnavus_group , Escherichia_Shigella , Lachnoclostridium , Klebsiella and other potential pathogens were detected in the fecal samples from UC patients, with a milder condition observed during the remission phase. Significant higher levels of corresponding SCFA were observed in the groups with addition of FOS-SCFAs esters during fermentation. Butyrylated fructooligosaccharides (B-FOS) and propionylated fructooligosaccharides (P-FOS) by enzymatic synthesis successfully promoted the proliferation of Bifidobacterium and inhibited Clostridium_sensu_stricto_1 and Klebsiella. Overall, B-FOS and P-FOS exhibit promising potential for restoring intestinal homeostasis and alleviating intestinal inflammation in individuals with UC. [Display omitted] [ABSTRACT FROM AUTHOR]

Propionic acid (PA), an end product of bacterial metabolism, has various functions in the colon. The aim of this study was to use ester bonds to link PA and a prebiotic fructooligosaccharides (FOS) to form propionyl-FOS capable of delivering PA to the colon. We used enzyme assistance and acid anhydride method to produce propionyl-FOS, respectively. FT-IR and NMR were used to confirm the formation of ester bonds. MALDI-TOF-MS was used to characterize the ester diversity and to partly monitor the fate of propionyl-FOS. In vitro simulated gut fermentation was used to study the effect of obtained esters on gut microbiota modulation and metabolite production. Our results showed that chemically synthesized propionyl-FOS (AP-FOS) was propionyl-esterified to a higher degree than enzymatically synthesized ones (P-FOS). Propionyl-fructooligosaccharides were more diverse in AP-FOS than in P-FOS. Higher levels of PA and butyric acid were detected in both AP-FOS and P-FOS groups than in FOS group after 24 h in vitro fermentation. The fermentability and prebiotic effects of AP-FOS were lower than those of P-FOS due to the high esterification degree of the former. Compared to FOS, P-FOS could confer better beneficial health effects through gut microbiota modulation. Hence, enzymatically synthesized P-FOS could be a potential prebiotic for health-promoting properties. [Display omitted] • Propionyl-FOS (P-FOS) has been successfully synthesized using an enzymatic method. • P-FOS could be well utilized by the human fecal microbiota. • P-FOS promoted SCFA production and reduced pH in simulated colonic fermentation. • P-FOS shaped the microbial community to a healthier pattern. • P-FOS could be a potential prebiotic for health-promoting properties. [ABSTRACT FROM AUTHOR]

Liver cancer is one of the leading causes of malignancy‑associated mortality worldwide and its clinical therapy remains very challenging. Ginsenoside Rh2 (Rh2) has been reported to have antitumor effects on some types of cancer, including liver cancer. However, its regulatory mechanism has not been extensively evaluated. In the present study, Rh2 increased the expression of microRNA (miR)‑200b‑5p, miR‑224‑3p and miR‑146a‑5p, and decreased the expression of miR‑26b‑3p and miR‑29a‑5p. Of the three upregulated miRs, miR‑146a‑5p exhibited the highest fold elevation. In accordance with a previous study, Rh2 effectively inhibited the survival of liver cancer cells in vitro and in a mouse model. In addition, it was observed that Rh2 markedly promoted liver cancer apoptosis and inhibited colony formation. Cell apoptosis and the inhibition of cell survival as well as colony formation induced by Rh2 were enhanced and weakened by miR‑146a‑5p overexpression and inhibition, respectively. The results of the present study provide further evidence of the antitumor effect of Rh2 in liver cancer and also demonstrate that this effect may be mediated via the regulation of miR‑146a‑5p expression in the liver cancer cell line HepG2. The results indicated that miR‑146a‑5p may be a promising regulatory factor in Rh2‑mediated effects in liver cancer. [ABSTRACT FROM AUTHOR]

Very long-chain fatty acid elongase 3 (ELOVL3) catalyzes the synthesis of C20–C24 fatty acids and is highly expressed in the liver and adipose tissues. The deficiency of Elovl3 exhibits an anti-obesity effect in mice, but the specific role of hepatic ELOVL3 in lipid metabolism remains unclear. Here we demonstrate that hepatic Elovl3 is not required for lipid homeostasis or the pathogenesis of diet-induced obesity and hepatic steatosis. We generated Elovl3 liver-specific knockout mice via Cre/LoxP approach, which maintained normal expression of ELOVL1 or ELOVL7 in the liver. Unexpectedly, the mutant mice did not show significant abnormalities in body weight, liver mass and morphology, liver triglyceride content, or glucose tolerance when fed normal chow or even a low-fat diet. Moreover, deletion of hepatic Elovl3 did not significantly affect body weight gain or hepatic steatosis induced by high-fat diet. Lipidomic analysis revealed that the lipid profiles were not significantly altered by the loss of hepatic Elovl3. Unlike its global knockouts, the mice lacking Elovl3 specifically in liver displayed normal expression of genes involved in hepatic de novo lipogenesis, lipid uptake, or beta-oxidation at the mRNA and protein levels. Collectively, our data indicate that hepatic ELOVL3 is dispensable for metabolic homeostasis or diet-induced metabolic disease. • We first used a tissue-specific Elovl3-knockout mouse model to investigate the function of liver ELOVL3. • Hepatic ELOVL3 is not required for lipid homeostasis. • Hepatic ELOVL3 is dispensable for the pathogenesis of diet-induced obesity and fatty liver. [ABSTRACT FROM AUTHOR]

• The hierarchical Ti 3 C 2 T X @NiO heterostructure was synthesized via chemical bath deposition along with thermal annealing. • By a GO-assisted self-convergence hydrothermal strategy, the Ti 3 C 2 T X @NiO was assembled into 3D Ti 3 C 2 T X @NiO-RGO hydrogel. • The 3D Ti 3 C 2 T X @NiO-RGO hydrogel with high mechanical strength provides excellent electrochemical performances. • An ultrahigh energy density supercapacitor device based on free-standing Ti 3 C 2 T X @NiO-RGO electrodes was assembled. 2D MXenes have been brought into sharp focus in electrochemical energy-storage devices due to their excellent electronic conductivity, high specific surface area, tunable layer structure, and large redox-active surface areas. Nevertheless, further applications of MXenes as electrode materials for supercapacitors are severely limited by the low theoretical specific capacity and self-restacking. Herein, a Ti 3 C 2 T X @NiO heterostructure is constructed firstly by uniformly deposition of NiO nanosheets on Ti 3 C 2 T X substrate, then the composite is incorporated into 3D porous hydrogel via an efficient graphene oxide (GO)-assisted self-convergence hydrothermal strategy with low temperatures. Owing to the synergistic effect among each components and the 3D porous interconnected architecture, the resultant 3D hierarchical Ti 3 C 2 T X @NiO-Reduced Graphene Oxide (RGO) heterostructure hydrogel not only takes both merits of highly conductive and outstanding pseudocapacitive, but also significantly prevents the aggregation and increases the surface utilization of Ti 3 C 2 T X @NiO. The 3D hierarchical Ti 3 C 2 T X @NiO-RGO heterostructure hydrogel electrodes (without binders) exhibit an enhanced electrochemical performance, with a superb specific capacitance up to 966 F g−1 at 1 A g−1 and long-term stability of 94.5% over 10,000 cycles. Importantly, a maximum energy density of 58.5 Wh kg−1 is obtained at power density of 550 W kg−1. Our work provides a facile and efficient approach to develop Ti 3 C 2 T X -based heterostructure hydrogels with improved overall performance, which expands the practical applications of MXenes in supercapacitors. [ABSTRACT FROM AUTHOR]

Concern is growing over the effects of neonicotinoid pesticides, which can impair honey bee cognition. We provide the first demonstration that sublethal concentrations of imidacloprid can harm honey bee decision-making about danger by significantly increasing the probability of a bee visiting a dangerous food source. Apis cerana is a native bee that is an important pollinator of agricultural crops and native plants in Asia. When foraging on nectar containing 40 µg/L (34 ppb) imidacloprid, honey bees (Apis cerana) showed no aversion to a feeder with a hornet predator, and 1.8 fold more bees chose the dangerous feeder as compared to control bees. Control bees exhibited significant predator avoidance. We also give the first evidence that foraging by A. cerana workers can be inhibited by sublethal concentrations of the pesticide, imidacloprid, which is widely used in Asia. Compared to bees collecting uncontaminated nectar, 23% fewer foragers returned to collect the nectar with 40 µg/L imidacloprid. Bees that did return respectively collected 46% and 63% less nectar containing 20 µg/L and 40 µg/L imidacloprid. These results suggest that the effects of neonicotinoids on honey bee decision-making and other advanced cognitive functions should be explored. Moreover, research should extend beyond the classic model, the European honey bee (A. mellifera), to other important bee species. [ABSTRACT FROM AUTHOR]

Background: Multimorbidity is common among fracture patients. However, its association with osteoporosis investigation and treatment to prevent future fractures is unclear. This limited knowledge impedes optimal patient care. This study investigated the association between multimorbidity and osteoporosis investigation and treatment in persons at high risk following an osteoporotic fracture. Methods and findings: The Sax Institute's 45 and Up Study is a prospective population-based cohort of 267,153 people in New South Wales, Australia, recruited between 2005 and 2009. This analysis followed up participants until 2017 for a median of 6 years (IQR: 4 to 8). Questionnaire data were linked to hospital admissions (Admitted Patients Data Collection (APDC)), emergency presentations (Emergency Department Data Collection (EDDC)), Pharmaceutical Benefits Scheme (PBS), and Medicare Benefits Schedule (MBS). Data were linked by the Centre for Health Record Linkage and stored in a secured computing environment. Fractures were identified from APDC and EDDC, Charlson Comorbidity Index (CCI) from APDC, Dual-energy X-ray absorptiometry (DXA) investigation from MBS, and osteoporosis treatment from PBS. Out of 25,280 persons with index fracture, 10,540 were classified as high-risk based on 10-year Garvan Fracture Risk (age, sex, weight, prior fracture and falls) threshold ≥20%. The association of CCI with likelihood of investigation and treatment initiation was determined by logistic regression adjusted for education, socioeconomic and lifestyle factors). The high-risk females and males averaged 77 ± 10 and 86 ± 5 years, respectively; >40% had a CCI ≥2. Only 17% of females and 7% of males received a DXA referral, and 22% of females and 14% males received osteoporosis medication following fracture. A higher CCI was associated with a lower probability of being investigated [adjusted OR, females: 0.73 (95% CI, 0.61 to 0.87) and 0.43 (95% CI, 0.30 to 0.62); males: 0.47 (95% CI, 0.33 to 0.68) and 0.52 (0.31 to 0.85) for CCI: 2 to 3, and ≥4 versus 0 to 1, respectively] and of receiving osteoporosis medication [adjusted OR, females: 0.85 (95% CI, 0.74 to 0.98) and 0.78 (95% CI, 0.61 to 0.99); males: 0.75 (95% CI, 0.59 to 0.94) and 0.37 (95% CI, 0.23 to 0.53) for CCI: 2 to 3, and ≥4 versus 0 to 1, respectively]. The cohort is relatively healthy; therefore, the impact of multimorbidity on osteoporosis management may have been underestimated. Conclusions: Multimorbidity contributed significantly to osteoporosis treatment gap. This suggests that fracture risk is either underestimated or underprioritized in the context of multimorbidity and highlights the need for extra vigilance and improved fracture care in this setting. Dana Bliuc and team investigate the association between multimorbidity and osteoporosis investigation and treatment in persons at high risk following an osteoporotic fracture. Author summary: Why was this study done?: Osteoporotic fractures are common, costly, and associated with increased risk of future fracture and premature mortality. Effective preventive treatment is available and recommended for high-risk patients with a 10-year fracture risk >20%, but its uptake in this group is suboptimal. High-risk patients often have multiple chronic conditions, and it is important to know to what degree the other chronic conditions might alter the uptake of fracture prevention medication in order to optimise patient care. What did the researchers do and find?: We conducted a prospective study including over 10,000 adults aged 50+ with an osteoporotic fracture and high-risk of future fracture to investigate the association between multimorbidity and osteoporosis investigation and treatment. In this high-risk group, the vast majority of adults were not investigated or treated for osteoporosis following an osteoporotic fracture. Adults with multiple chronic conditions were significantly less likely to be investigated [adjusted OR, females: 0.73 (95% CI, 0.61 to 0.87) and 0.43 (95% CI, 0.30 to 0.62); males: 0.47 (95% CI, 0.33 to 0.68) and 0.52 (0.31 to 0.85) for CCI: 2 to 3, and ≥4 versus 0 to 1, respectively] or treated for osteoporosis [adjusted OR, females: 0.85 (95% CI, 0.74 to 0.98) and 0.78 (95% CI, 0.61 to 0.99); males: 0.75 (95% CI, 0.59 to 0.94) and 0.37 (95% CI, 0.23 to 0.53) for CCI: 2 to 3, and ≥4 versus 0 to 1, respectively] compared to those without chronic conditions. What do these findings mean?: The presence of additional chronic conditions was associated with a lower likelihood of being investigated and treated for osteoporosis. These findings suggest that fracture preventing treatment is either underprioritised or underestimated in the presence of other chronic conditions. There is a need for more awareness of increased fracture risk in complex patients with multiple chronic conditions. [ABSTRACT FROM AUTHOR]

Background: Cassava (Manihot esculenta Crantz) is an important multiuse crop grown for economic and energy purposes. Its vegetative organs are storage roots, in which the main storage material is starch. The accumulation characteristics of starch in cassava roots can directly affect the yield, starch content and maturation of cassava storage roots. In this study, we used a cassava sexual tetraploid (ST), which showed early maturation heterosis in previous work, as the main test material. We analyzed the sucrose metabolism and starch accumulation characteristics of the ST and its parents from the leaf "source" to the storage root "sink" during different developmental stages and explored the regulatory mechanisms of ST storage root early maturation by combining the transcriptome data of the storage roots during the expansion period. Results: The results showed that the trends in sucrose, glucose and fructose contents in the ST leaves were similar to those of the two parents during different stages of development, but the trends in the ST storage roots were significantly different from those of their parents, which showed high sucrose utilization rates during the early stage of development and decreased utilization capacity in the late developmental stage. Transcriptome data showed that the genes that were expressed differentially between ST and its parents were mainly involved in the degradation and utilization of sucrose in the storage roots, and four key enzyme genes were significantly upregulated (Invertase MeNINV8/MeVINV3, Sucrose synthase MeSuSy2, Hexokinase MeHXK2), while the expressions of key enzyme genes involved in starch synthesis were not significantly different. Conclusions: The results revealed that the pattern of sucrose degradation and utilization in the cassava ST was different from that of its parents and promoted early maturation in its tuberous roots. Starch accumulation in the ST from sucrose mainly occurred during the early expansion stage of the storage roots, and the starch content during this period was higher than that of both parents, mainly due to the regulation of invertase and hexokinase activities during sucrose metabolism. This study provides a basis for further genetic improvements to cassava traits and for breeding varieties that mature early and are adapted well to provide starch supply requirements. [ABSTRACT FROM AUTHOR]

Stannic tetrachloride was an efficient Lewis acid catalyst for the aza-acetalization of aromatic aldehydes with o-arylaminomethyl phenols, and a series of novel aryl substituted 3,4-dihydro-2H-1,3-benzoxazines were prepared in good yields under mild conditions. SnCl4 was a more efficient catalyst for the reaction than p-toluenesulfonic acid, sulfuric acid, and aluminium chloride. [ABSTRACT FROM AUTHOR]

Abstract: We investigated the implicit learning of a linguistically relevant variable (animacy) in a natural language context (namely, the relation of forms of determiners to semantics). Trial by trial subjective measures indicated that exposure to a form–animacy regularity led to unconscious knowledge of that regularity. Under the same conditions, people did not learn about another form–meaning regularity when a linguistically arbitrary variable was used instead of animacy (size relative to a dog). Implicit learning is constrained to acquire unconscious knowledge about features with high prior probabilities of being relevant in that domain. [Copyright &y& Elsevier]

Background: Endocrine resistance driven by sustained activation of androgen receptor (AR) signaling pathway in advanced prostate cancer (PCa) is fatal. Characterization of mechanisms underlying aberrant AR pathway activation to search for potential therapeutic strategy is particularly important. Rac GTPase-activating protein 1 (RACGAP1) is one of the specific GTPase-activating proteins. As a novel tumor proto-oncogene, overexpression of RACGAP1 was related to the occurrence of various tumors. Methods: Bioinformatics methods were used to analyze the relationship of expression level between RACGAP1 and AR as well as AR pathway activation. qRT-PCR and western blotting assays were performed to assess the expression of AR/AR-V7 and RACGAP1 in PCa cells. Immunoprecipitation and immunofluorescence experiments were conducted to detect the interaction and co-localization between RACGAP1 and AR/AR-V7. Gain- and loss-of-function analyses were conducted to investigate the biological roles of RACGAP1 in PCa cells, using MTS and colony formation assays. In vivo experiments were conducted to evaluate the effect of RACGAP1 inhibition on the tumor growth. Results: RACGAP1 was a gene activated by AR, which was markedly upregulated in PCa patients with CRPC and enzalutamide resistance. AR transcriptionally activated RACGAP1 expression by binding to its promoter region. Reciprocally, nuclear RACGAP1 bound to the N-terminal domain (NTD) of both AR and AR-V7, blocking their interaction with the E3 ubiquitin ligase MDM2. Consequently, this prevented the degradation of AR/AR-V7 in a ubiquitin-proteasome-dependent pathway. Notably, the positive feedback loop between RACGAP1 and AR/AR-V7 contributed to endocrine therapy resistance of CRPC. Combination of enzalutamide and in vivo cholesterol-conjugated RIG-I siRNA drugs targeting RACGAP1 induced potent inhibition of xenograft tumor growth of PCa. Conclusion: In summary, our results reveal that reciprocal regulation between RACGAP1 and AR/AR-V7 contributes to the endocrine resistance in PCa. These findings highlight the therapeutic potential of combined RACGAP1 inhibition and enzalutamide in treatment of advanced PCa. [ABSTRACT FROM AUTHOR]

Background: Parkinson's disease (PD) is a progressive neurodegenerative disease with increasing prevalence. Effective diagnostic markers and therapeutic methods are still lacking. Exploring key molecular markers and mechanisms for PD can help with early diagnosis and treatment improvement. Methods: Three datasets GSE174052, GSE77668, and GSE168496 were obtained from the GEO database to search differentially expressed circRNA (DECs), miRNAs (DEMis), and mRNAs (DEMs). GO and KEGG enrichment analyses, and protein–protein interaction (PPI) network construction were implemented to explore possible actions of DEMs. Hub genes were selected to establish circRNA-related competing endogenous RNA (ceRNA) networks. Results: There were 1005 downregulated DECs, 21 upregulated and 21 downregulated DEMis, and 266 upregulated and 234 downregulated DEMs identified. The DEMs were significantly enriched in various PD-associated functions and pathways such as extracellular matrix organization, dopamine synthesis, PI3K-Akt, and calcium signaling pathways. Twenty-one hub genes were screened out, and a PD-related ceRNA regulatory network was constructed containing 31 circRNAs, one miRNA (miR-371a-3p), and one hub gene (KCNJ6). Conclusion: We identified PD-related molecular markers and ceRNA regulatory networks, providing new directions for PD diagnosis and treatment. [ABSTRACT FROM AUTHOR]

• Ligands of Nl Gr10a are fructose and cellobiose, a ligand of Nl Gr10b is arabinose. • Nl Gr10a modulates fecundity through Nl Vg and Nl VgR, but Nl Gr10b only through Nl Vg. • Nl Gr10a and Nl Gr10b modulates fecundity through AMPK and AKT signaling pathways. Insect gustatory receptors play crucial roles in multiple physiological behaviours. Although the alternative splicing of some gustatory receptors has been observed in insect species, differences in their ligands and functions have rarely been reported. Here, we cloned NlGr10a and NlGr10b , two alternative splicing variants of a sugar gustatory receptor gene in the brown planthopper (BPH), Nilaparvata lugens (Stål), and found that their ligands were different by calcium imaging assay. The ligands of Nl Gr10a were fructose and cellobiose, and the ligand of Nl Gr10b was arabinose. Subsequently, the RNAi results showed that knockdown of both splicing variants decreased the number of eggs laid by BPH females, and the egg hatching rate after knockdown of NlGr10a was significantly lower than that after knockdown of NlGr10b. Furthermore, Nl Gr10a promoted the fecundity of BPH through the AMPK- and AKT- Nl Vg/ Nl VgR signalling pathways, whereas Nl Gr10b promoted the fecundity only through the AMPK- and AKT- Nl Vg signalling pathways. These findings broaden our understanding of the difference in the ligands and functions of alternative splicing variants of gustatory receptors in insects. [ABSTRACT FROM AUTHOR]

Existing prebiotics, such as fructo-oligosaccharides (FOSs), can be modified to enhance their functionality or introduce additional functionalities. This study aimed to investigate the fermentation characteristics and prebiotic potential of enzymatically synthesized butyryl-FOSs. The esters were successfully synthesized through the reaction of butyric acid and FOSs using both chemical and enzymatic methods, denoted as A-FOSs and B-FOSs, respectively, for comparative analysis. The esterification degree of each component in A-FOSs was significantly higher than that of B-FOSs. Subsequently, the obtained esters were characterized for their fermentation properties, degradation mode and potential prebiotic effects using an in vitro simulated colonic fermentation model. Enzymes of human gut microbiota were found to preferentially cleave the glycosidic bond to the unit without butyryl group and release the sugars for utilization. A significant increase in butyric acid levels was observed during fermentation after the supplementation of B-FOSs. The 16S rRNA gene sequencing, absolute quantification of microbiota, and selected probiotic strains culture showed that B-FOSs supplementation promoted the growth of beneficial bacteria while reducing harmful ones. These results suggest that B-FOSs hold promise as novel prebiotics, possessing dual functions of modulating gut microbiota and delivering butyric acid to the colon in a targeted manner, ultimately contributing to improved gut health. [Display omitted] [ABSTRACT FROM AUTHOR]

Background: Psoriasis (PA) is a chronic autoimmune disease of the skin that adversely affects patients' quality of life. Yangxue Jiedu Fang (YXJD) has been used for decades to treat psoriasis in China. However, its antipsoriatic mechanisms are still poorly understood. In this study, we explored the effects of YXJD on angiogenesis and apoptosis of microvessels in PA, the underlying mechanisms in HUVEC cells transfected by Survivin overexpression plasmid and in a mouse model of imiquimod-induced psoriasis and the relationship between VEGF (vascular endothelial growth factor) and Survivin.Methods: A BALB/c mouse model of imiquimod- (IMQ-) induced PA was established, and the mice were treated with YXJD. Cell viability was assessed by CCK8 assay. Apoptosis was detected by annexin V-FITC/PI double-staining and caspase-3 assays. The PI3K/Akt/β-catenin pathway was analyzed by western blotting, ELISA, and immunochemical analysis.Results: YXJD ameliorated symptoms and psoriasis area and severity index (PASI) scores and also reduced the number of microvessels, as determined by the microvessel density (MVD). The expression of apoptotic protein Survivin in endothelial cells, autophagy-related proteins p62, and angiogenic proteins VEGF was inhibited by YXJD, and the repressed expression of LC3II/I increased by YXJD. The proteins related to the PI3K/Akt pathway and β-catenin expression and the nuclear entry of β-catenin were reduced in IMQ-induced PA mice treated with YXJD. In HUVEC cells transfected by Survivin overexpression plasmid, we observed YXJD regulated the expression of Survivin, LC3II/I, and p62, VEGF, and PI3K/Akt pathway-relative proteins and the nuclear entry of β-catenin.Conclusions: YXJD inhibited the expression of Survivin via PI3K/Akt pathway to adjust apoptosis, autophagy, and angiogenesis of microvessels and thus improve the vascular sustainability in psoriasis. YXJD may represent a new direction of drug research and development for immunomodulatory therapy for psoriasis. [ABSTRACT FROM AUTHOR]

Phosphatase and tensin homolog (PTEN) loss tightly correlates with prostate cancer (PCa) progression and metastasis. Inactivation of PTEN leads to abnormal activation of PI3K/AKT pathway. However, results from clinical trials with AKT inhibitors in PCa have been largely disappointing. Identification of novel regulators of PTEN in PTEN-dysfunctional PCa is urgently needed. Here we demonstrated that the expression level of PTEN is inversely correlated with the signature score of unfolded protein response (UPR) in PCa. Importantly, PTEN suppresses the activity of ATF6α, via interacting to de-phosphorylate ATF6α and consequently inhibiting its nuclear translocation. Conversely, ATF6α promotes the ubiquitination and degradation of PTEN by inducing CHIP expression. Thus, ATF6α and PTEN forms a negative feedback loop during PCa progression. Combination of ATF6α inhibitor with AKT inhibitor suppresses tumor cell proliferation and xenograft growth. Importantly, this study highlighted ATF6α as a therapeutic vulnerability in PTEN dysfunctional PCa. [ABSTRACT FROM AUTHOR]

Background: Castration-resistant prostate cancer (CRPC) is currently the main challenge for prostate cancer (PCa) treatment, and there is an urgent need to find novel therapeutic targets and drugs. Prohibitin (PHB1) is a multifunctional chaperone/scaffold protein that is upregulated in various cancers and plays a pro-cancer role. FL3 is a synthetic flavagline drug that inhibits cancer cell proliferation by targeting PHB1. However, the biological functions of PHB1 in CRPC and the effect of FL3 on CRPC cells remain to be explored. Methods: Several public datasets were used to analyze the association between the expression level of PHB1 and PCa progression as well as outcome in PCa patients. The expression of PHB1 in human PCa specimens and PCa cell lines was examined by immunohistochemistry (IHC), qRT-PCR, and Western blot. The biological roles of PHB1 in castration resistance and underlying mechanisms were investigated by gain/loss-of-function analyses. Next, in vitro and in vivo experiments were conducted to investigate the anti-cancer effects of FL3 on CRPC cells as well as the underlying mechanisms. Results: PHB1 expression was significantly upregulated in CRPC and was associated with poor prognosis. PHB1 promoted castration resistance of PCa cells under androgen deprivation condition. PHB1 is an androgen receptor (AR) suppressive gene, and androgen deprivation promoted the PHB1 expression and its nucleus-cytoplasmic translocation. FL3, alone or combined with the second-generation anti-androgen Enzalutamide (ENZ), suppressed CRPC cells especially ENZ-sensitive CRPC cells both in vitro and in vivo. Mechanically, we demonstrated that FL3 promoted trafficking of PHB1 from plasma membrane and mitochondria to nucleus, which in turn inhibited AR signaling as well as MAPK signaling, yet promoted apoptosis in CRPC cells. Conclusion: Our data indicated that PHB1 is aberrantly upregulated in CRPC and is involved in castration resistance, as well as providing a novel rational approach for treating ENZ-sensitive CRPC. [ABSTRACT FROM AUTHOR]

Xenobiotics such as the neonicotinoid pesticide, imidacloprid, are used globally, but their effects on native bee species are poorly understood. We studied the effects of sublethal doses of imidacloprid on olfactory learning in the native honey bee species, Apis cerana, an important pollinator of agricultural and native plants throughout Asia. We provide the first evidence that imidacloprid can impair learning in A. cerana workers exposed as adults or as larvae. Adults that ingested a single imidacloprid dose as low as 0.1 ng/bee had significantly reduced olfactory learning acquisition, which was 1.6-fold higher in control bees. Longer-term learning (1-17 h after the last learning trial) was also impaired. Bees exposed as larvae to a total dose of 0.24 ng/bee did not have reduced survival to adulthood. However, these larval-treated bees had significantly impaired olfactory learning when tested as adults: control bees exhibited up to 4.8-fold better short-term learning acquisition, though longer-term learning was not affected. Thus, sublethal cognitive deficits elicited by neonicotinoids on a broad range of native bee species deserve further study. [ABSTRACT FROM AUTHOR]

Background. Jinshuibao capsules (JSB) have been widely used to treat early diabetic nephropathy (DN), but the specific effects are still inconsistent. A meta-analysis of randomized controlled trials (RCTs) was conducted to evaluate the clinical efficacy of JSB for early DN. Methods. Four international databases and four Chinese databases were searched from publication dates to March 1, 2018. The RCTs reporting the results of JSB’s specific effects were included, and comparisons were between JSB combined with Angiotensin Receptor Blockers (ARBs) as experimental intervention and ARBs as the control. Included studies’ quality was evaluated and the extracted data were analyzed with RevMan 5.3 software. Results. Twenty-six RCTs including 2198 early DN participants were adopted in the meta-analysis. The results showed that, compared with the ARBs alone, JSB could remarkably improve the ORR (OR = 3.84; 95% CI: 2.37~6.24; P<0.00001) and decrease 24 h UTP (MD = −93.32; 95% CI: −128.60 ~-58.04; P<0.00001), UAER (MD = −24.02; 95% CI: −30.93 ~-17.11; P<0.00001), BUN (MD = −0.26; 95%: −0.44 ~-0.08; P=0.005), Scr (MD = −9.07; 95% CI: −14.26 ~-3.88; P=0.0006), ACR (MD = −17.55; 95% CI: −22.81 ~-12.29; P<0.00001), Cys-C (MD = −0.60; 95% CI: −0.88 ~-0.32; P<0.00001), SBP (MD = −3.08; 95% CI: −4.65 ~-1.52; P=0.0001), DBP (MD = −2.09; 95% CI: −4.00 ~-0.19; P=0.03), and TG (MD = −0.36; 95% CI: −0.50 ~-0.21; P<0.00001). However, it showed no significant differences in TC (MD = −0.32; 95% CI: −0.69~0.04; P=0.08), FBG (MD = 0.04; 95% CI: −0.39~0.47; P=0.87), HbA1c (MD = −0.26; 95% CI: −0.59~0.06; P=0.11), and β2-MG (MD = −15.61; 95% CI: −32.95~1.73; P=0.08). Conclusions. This study indicates that JSB is an effective accessory therapeutic medicine for patients with early DN. It contributes to decreasing blood pressure and the content of triglyceride and improving the renal function of early DN patients. However, there is still a need to further verify the auxiliary therapeutic effect of JSB with more strictly designed RCTs with large sample and multiple centers in the future. [ABSTRACT FROM AUTHOR]

We combined pyropheophorbide-a methyl ester, photodynamic therapy (MPPa-PDT), 670 ± 10 nm, 4 mW/cm2, with herpes simplex virus type 1 thymidine kinase/ganciclovir (HSV1-TK/GCV) to improve the therapeutic effect. We built HSV1-TK expression vector GV230-TK and we observed a bright green fluorescence under fluorescence microscope. It indicated the recombinant plasmid was transfected into PC-3M prostate cancer cells successfully. As the abundant glucose-regulated protein 78 (GRP78) promoter in PC-3M cells can cause active expression of HSV1-TK, cell protein was collected for western blot to determine the expression of HSV1-TK. In CCK-8 assay (n = 6), the cell survival rate of combined treatment group was about 10%, less than that of pure MPPa-PDT group (23%) and pure HSV1-TK/GCV group (35%) (t test, P < 0.05). Flow cytometry was used to measure the cytotoxicity; the apoptosis rate of combined treatment group was about 38%, higher than that of pure MPPa-PDT group (about 22%) and pure HSV1-TK/GCV group (about 19%). The results showed that the combination of the two treatments can effectively improve the cytocidal effect in PC-3M cells. [ABSTRACT FROM AUTHOR]

Plants produce diverse secondary compounds as natural protection against microbial and insect attack. Most of these compounds, including bitters and acids, are sensed by insect gustatory receptors (Grs). Although some organic acids are attractive at low or moderate levels, most acidic compounds are potentially toxic to insects and repress food consumption at high concentrations. At present, the majority of the reported sour receptors function in appetitive behaviors rather than aversive taste responses. Here, using two different heterologous expression systems, the insect Sf9 cell line and the mammalian HEK293T cell line, we started from crude extracts of rice (Oryza sativa) and successfully identified oxalic acid (OA) as a ligand of NlGr23a, a Gr in the brown planthopper Nilaparvata lugens that feeds solely on rice. The antifeedant effect of OA on the brown planthopper was dose dependent, and NlGr23a mediated the repulsive responses to OA in both rice plants and artificial diets. To our knowledge, OA is the first identified ligand of Grs starting from plant crude extracts. These findings on rice–planthopper interactions will be of broad interest for pest control in agriculture and also for better understanding of how insects select host plants. [ABSTRACT FROM AUTHOR]

Insect fecundity is a quantitative phenotype strongly affected by genotypes and the environment. However, interactions between genotypes and environmental factors in modulating insect fecundity remain largely unknown. This study investigated the impact of population density on the fecundity of Nilaparvata lugens (brown planthopper; BPH) carrying homozygous high‐ (HFG) or low‐ (LFG) fecundity homozygous genotypes. Under low population densities, the fecundity and population growth rate of both genotypes showed similar increasing trends across generations, while the trends between HFG and LFG under high population densities were opposite. Through a combination of temporal analysis and weighted gene co‐expression network analyses on RNA‐seq data of HFG and LFG under low and high population densities in the 1st, 3rd, and 5th generations, we identified 2 gene modules that were associated with these density‐dependent progenitive phenotypes. Four pathways related to the neural system were simultaneously enriched by the 2 gene modules. Furthermore, Nlpale, which encodes a tyrosine hydroxylase, was identified as a key gene. The RNA interference of this gene and manipulation of its downstream product dopamine significantly affected the basic and density‐dependent progenitive phenotypes of BPH. These findings indicated that dopamine biosynthesis is the key regulatory factor that determines fecundity in response to density changes in different BPH genotypes. Thus, this study provides insights into the interaction of a typical environmental factor and insect genotype during the process of population regulation. [ABSTRACT FROM AUTHOR]

In this study, we demonstrate an approach for inverting earthquake source parameters based on high-rate global positioning system (GPS) velocity seismograms. The velocity records obtained from single-station GPS velocity solutions with broadcast ephemeris are used directly for earthquake source parameter inversion using the Cut and Paste method, without requiring conversion of the velocity records into displacement records. Taking the El Mayor-Cucapah earthquake as an example, GPS velocity records from 10 stations with reasonable azimuthal coverage provide earthquake source parameters very close to those from the Global centroid moment tensor (Global CMT) solution. In sparse network tests, robust source parameters with acceptable bias can be achieved with as few as three stations. When the number of stations is reduced to two, the bias in rake angle becomes appreciable, but the magnitude and strike estimations are still robust. The results of this study demonstrate that rapid and reliable estimation of earthquake source parameters can be obtained from GPS velocity data. These parameters could be used for early earthquake warning and shake map construction, because such GPS velocity records can be obtained in real time. [ABSTRACT FROM AUTHOR]

Background: The development of castration-resistant prostate cancer (CRPC) remains a major obstacle in the treatment of prostate cancer (PCa). Dysregulated mitochondrial function has been linked to the initiation and progression of diverse human cancers. Deciphering the novel molecular mechanisms underlying mitochondrial function may provide important insights for developing novel therapeutics for CRPC. Methods: We investigate the expression of the protein tyrosine phosphatase receptor type F polypeptide interacting protein alpha 4 (PPFIA4) using public datasets and tumor specimens from PCa cases by immunohistochemistry. Gain- and loss-of-function studies are performed in PCa cell lines and mouse models of subcutaneous xenograft to characterize the role of PPFIA4 in CRPC. Gene expression regulation is evaluated by a series of molecular and biochemical experiments in PCa cell lines. The therapeutic effects of methylenetetrahydrofolate dehydrogenase 2 (MTHFD2) inhibitor combined enzalutamide are assessed using in vitro functional assays and in vivo mouse models. Results: We show that the increase of PPFIA4 exacerbates aggressive phenotype resembling CRPC. A fraction of PPFIA4 localizes to mitochondria and interacts with MTHFD2, a key enzyme for one-carbon metabolism. Androgen deprivation increases the translocation of PPFIA4 into mitochondria and increases the interaction between PPFIA4 and MTHFD2, which result in the elevation of tyrosine phosphorylated MTHFD2. Consequently, the levels of NADPH synthesis increase, resulting in protection against androgen deprivation-induced mitochondrial dysfunction, as well as promotion of tumor growth. Clinically, PPFIA4 expression is significantly increased in CRPC tissues compared with localized PCa ones. Importantly, an MTHFD2 inhibitor, DS18561882, combined with enzalutamide can significantly inhibit CRPC cell proliferation in vitro and tumor growth in vivo. Conclusion: Overall, our findings reveal a PPFIA4-MTHFD2 complex in mitochondria that links androgen deprivation to mitochondrial metabolism and mitochondrial dysfunction, which suggest a potential strategy to inhibit CRPC progression. [ABSTRACT FROM AUTHOR]

Fear can have strong ecosystem effects by giving predators a role disproportionate to their actual kill rates. In bees, fear is shown through foragers avoiding dangerous food sites, thereby reducing the fitness of pollinated plants. However, it remains unclear how fear affects pollinators in a complex natural scenario involving multiple predator species and different patch qualities. We studied hornets, Vespa velutina (smaller) and V. tropica (bigger) preying upon the Asian honey bee, Apis cerana in China. Hornets hunted bees on flowers and were attacked by bee colonies. Bees treated the bigger hornet species (which is 4 fold more massive) as more dangerous. It received 4.5 fold more attackers than the smaller hornet species. We tested bee responses to a three-feeder array with different hornet species and varying resource qualities. When all feeders offered 30% sucrose solution (w/w), colony foraging allocation, individual visits, and individual patch residence times were reduced according to the degree of danger. Predator presence reduced foraging visits by 55–79% and residence times by 17–33%. When feeders offered different reward levels (15%, 30%, or 45% sucrose), colony and individual foraging favored higher sugar concentrations. However, when balancing food quality against multiple threats (sweeter food corresponding to higher danger), colonies exhibited greater fear than individuals. Colonies decreased foraging at low and high danger patches. Individuals exhibited less fear and only decreased visits to the high danger patch. Contrasting individual with emergent colony-level effects of fear can thus illuminate how predators shape pollination by social bees. [ABSTRACT FROM AUTHOR]

[Display omitted] • 1 Whey protein (WP)/FOS microencapsulation enhances anthocyanin antioxidant activity. • 2 WP/FOS microencapsulation increases anthocyanin stability during in vitro digestion. • 3 WP/FOS-based anthocyanin microcapsules show improved fermentability. • 4 WP/FOS-based anthocyanin microcapsules positively impact microbiota composition. • 5 WP/FOS-based anthocyanin microcapsules significantly boost SCFA production. Anthocyanins are the primary functional pigments in the diet. However, anthocyanins exhibit instability during digestion, coupled with limited bioavailability. Microencapsulation offers anthocyanins a sheltered environment, enhancing their stability and bioactivity. Fructooligosaccharides (FOS) and whey protein (WP) commonly serve as wall materials in microencapsulation and represent a significant source of probiotic functionality. Our prior research successfully established a robust microencapsulation system for anthocyanins utilizing FOS and WP. This study investigates the antioxidative capacity, stability during in vitro digestion, modulation on gut microbiota, and short-chain fatty acids (SCFAs) production of black soybean skin anthocyanins microencapsulated with FOS and WP (anthocyanin-loaded microencapsule particles, ALM). The results demonstrate that ALM exhibits a superior antioxidant capacity compared to free anthocyanins (ANCs) and cyanidin-3-glucoside (C3G). During simulated digestion, ALM exhibits enhanced anthocyanin retention compared with ANC in both gastric and intestinal phases. In comparison with ANC and even non-loaded microcapsules (NLM), in vitro fermentation demonstrates that ALM exhibits the highest gas production and lowered pH, indicating excellent fermentation activity. Furthermore, in comparison with ANC or NLM, ALM exerts a positive influence on the diversity and composition of gut microbiota, with potentially beneficial genera such as Faecalibacterium and Akkermansia exhibiting higher relative abundance. Moreover, ALM stimulates the production of SCFAs, particularly acetic and propionic acids. In conclusion, microencapsulation of anthocyanins with FOS-WP enhances their antioxidative capacity and stability during in vitro digestion. Simultaneously, this microencapsulation illustrates a positive regulatory effect on the intestinal microbiota community and SCFA production, conferring potential health benefits. [ABSTRACT FROM AUTHOR]

Gentiopicroside (GPS), a main active secoiridoid glucoside derived from the roots of perennial herbs in the Gentianaceae family, has antispasmodic and relaxant effects. However, the vasorelaxant effects of GPS on aortic rings and the molecular mechanisms involved in these effects are not yet clear. Therefore, we investigated whether GPS inhibits phenylephrine- (PE-) or KCl-induced contractions in isolated rat thoracic aortic rings. The present study found that GPS produced a dose-dependent relaxation in aortic rings precontracted with PE or KCl and significantly reduced CaCl2-, narciclasine- (Rho-kinase activator-), and phorbol-12,13-diacetate- (PKC activator-) induced vasocontractions. Pretreatment with NG-Nitroarginine methyl ester hydrochloride (L-NAME, NOS inhibitor), methylene blue (sGC inhibitor), indomethacin (COX inhibitor), 4-aminopyridine (KV channel inhibitor), and glibenclamide (KATP channel inhibitor) had no influence on the vasorelaxant effect of GPS, while BaCl2 (Kir channel inhibitor), tetraethylammonium chloride (KCa channel inhibitor), ruthenium red (RYR inhibitor), and heparin (IP3R inhibitor) significantly reduced GPS-induced vasorelaxation. Moreover, GPS pretreatment remarkably inhibited the influx of Ca2+ in vascular smooth muscle cells stimulated using KCl or PE-containing CaCl2 solution. Western blot analysis confirmed that GPS treatment inhibited PE-induced increases in the protein levels of p-Akt, p-myosin light chain (MLC), and p-myosin-binding subunit of myosin phosphatase 1 (MYPT1) in the aortic rings. Additionally, the vasorelaxation activity of GPS was attenuated upon pretreatment with LY294002 (PI3K/Akt inhibitor), Y27632 (Rho-kinase inhibitor), and verapamil (L-type Ca2+ channel inhibitor). These findings demonstrate that GPS exhibits endothelium-independent vasorelaxant effects through inhibition of voltage-dependent, receptor-operated, and inositol triphosphate receptor (IP3R)/ryanodine receptor- (RYR-) mediated Ca2+ channels as well as the PI3K/Akt/Rho-kinase signaling pathway. [ABSTRACT FROM AUTHOR]

Metformin is commonly used, but approximately 20% of patients experience gastrointestinal intolerance, leading to medication discontinuation for unclear reasons and a lack of effective management strategies. In this study, the 18 fecal and blood samples were analyzed using 16S rRNA and mRNA transcriptome, respectively. These samples included 3 fecal and 4 blood from metformin-tolerant T2D patients before and after metformin treatment (T and Ta), 3 fecal and 5 blood from metformin-intolerant T2D patients before and after treatment (TS and TSa), and 6 fecal samples from healthy controls. The results showed that certain anti-inflammatory gut bacteria and gene, such as Barnesiella (p = 0.046), Parabacteroides goldsteinii (p = 0.016), and the gene JUND (p = 0.0002), exhibited higher levels in metformin-intolerant patients, and which decreased after metformin treatment (p < 0.05). This potentially invalidates patients' anti-inflammatory effect and intestinal mucus barrier protection, which may lead to alterations in intestinal permeability, decreased gut barrier function, and gastrointestinal symptoms, including diarrhea, bloating, and nausea. After metformin treatment, primary bile acids (PBAs) production species: Weissella confusa , Weissella paramesenteroides , Lactobacillus brevis , and Lactobacillus plantarum increased (p < 0.05). The species converting PBAs to secondary bile acids (SBAs): Parabacteroides distasonis decreased (p < 0.05). This might result in accumulation of PBAs, which also may lead to anti-inflammatory gene JUND and SQSTM1 downregulated. In conclusion, this study suggests that metformin intolerance may be attributed to a decrease in anti-inflammatory-related flora and genes, and also alterations in PBAs accumulation-related flora. These findings open up possibilities for future research targeting gut flora and host genes to prevent metformin intolerance. • Metformin intolerance mechanism in Chinese individuals may unique. • Intolerant subjects show a decline in anti-inflammatory gut bacteria and genes. • Gut flora changes in intolerant subjects may cause an excess of primary bile acids. • Inflammation and primary bile acids-related flora and genes may impact intolerance. [ABSTRACT FROM AUTHOR]

We map the distribution of macroseismic intensities from the M W 6.9 Kani and the M W 6.8 Chauk intra-slab earthquakes in 2016 in Myanmar using the 1998 European Macroseismic Scale (EMS-98) by interpreting data gathered from field surveys, community responses sent via social media to the Myanmar Earthquake Committee (MEC), and digital news reports. Our macroseismic maps for both events provide better spatial data coverage in Myanmar, India, and Bangladesh than community derived macroseismic maps (e.g., U.S. Geological Survey's "Did You Feel It?"). In Myanmar, this was driven by improved telecommunication that has allowed social media such as the Burmese language Facebook portal of the Myanmar Earthquake Committee (MEC) to reach into rural areas from where reports of shaking effects from earthquakes have been previously unavailable. Our analysis of both the macroseismic intensities and strong motion observations from India and Myanmar suggests the two earthquakes had different source properties. The comparison of our intensity data with instrumental strong motion records also suggests the peak ground motion-intensity relationship by Worden et al. (2012) generally performs well for both earthquakes. In addition, ground motion behaviour within the Burma and Indian plates can be related to different existing ground motion prediction equations (GMPEs) and intensity prediction equations (IPEs) for subduction zones and for stable continental regions respectively. We therefore suggest these effects will need to be considered in future regional seismic hazard models or Shake Maps for this region when evaluating the impact of the future events. • Transfrontier macroseismic intensity for 2016 Kani and Chauk intra-slab earthquakes in Myanmar • Analysis of strong motion data from Myanmar, India, and Thailand • Source and rupture directivity properties varied for both the Kani and Chauk earthquakes. • Preliminary observations of different grades of damage to pagodas with epicentral distance • Preliminary analysis of GMPE's, IPE's and GMICE for use in Myanmar [ABSTRACT FROM AUTHOR]

• Transcriptomic analysis from five different tissues reveal candidate unigenes involved in flavonoid metabolism. • Flavonoid profiles show the accumulation of the active ingredients in Chrysanthemum indicum. • One flavone synthases (FNSⅡ) was identified which catalyzes the conversion of flavanones to flavones. • A pharmacological component-linarin biosynthesis pathway is proposed. • Lay a foundation of potentials for molecular breeding and metabolic engineering in C. indicum. Chrysanthemum indicum L. is a type of herb that is widely used in China, Korea, and Japan. It has been used as an ingredient in traditional medicines, tea, and functional food because of its various anti-inflammatory and anti-oxidant bioactivities. Such bioactivities have been associated with flavonoids such as apigenin, luteolin and linarin in C. indicum. However, the biosynthesis pathway has not been investigated. In this study, using transcriptomic analysis and targeted metabolic profiling from five different tissues, we characterize the levels of flavonoids and mine the corresponding genes involved in flavonoid biosynthesis. Transcriptomic analysis revealed that 103 unigenes are involved in flavonoid-related biosynthesis pathways. Flavone synthase (FNS) is the key enzyme responsible for flavone synthesis and provides precursors for acacetin and linarin biosynthesis. One putative FNS Ⅱ gene, with the highest Reads Per Kilobase per Million mapped reads (RPKM) in flower and flower bud was cloned. Quantitative real-time polymerase chain reaction (RT-qPCR) revealed that CiFNSⅡ exhibited a similar expression pattern to that in the transcriptome in terms of RPKM. In addition, a targeted metabolic profiling of three flavanones (naringenin, eriodictyol, and liquiritigenin), three flavones (apigenin, luteolin, and 7,4′-dihydroxyflavone), and two flavone derivatives (linarin and acacetin) was performed to characterize the distribution of these flavonoids in different tissues of C. indicum. The recombinant FNSⅡ protein expressed in yeast was able to catalyze the conversion of three flavanones into the respective flavones. Based on the transcriptome analysis, metabolic profiling, and activity assays, a linarin biosynthesis pathway is proposed. Our study provides insight into the potential application of molecular breeding and metabolic engineering for improving the quality of cultivated C. indicum. [ABSTRACT FROM AUTHOR]

We aimed to study the structural characteristics and fermentation properties of wheat bran cell wall polysaccharides (CWPs). Sequential extractions of CWPs from wheat bran produced the water-extractable (WE) and alkali-extractable (AE) fractions. The extracted fractions were structurally characterized based on their molecular weight (Mw) and monosaccharide composition. Our findings revealed that the Mw and the ratio of arabinose to xylose (A/X) of AE were higher than those of WE and that the two fractions were mainly composed of arabinoxylans (AXs). The substrates were then subjected to in vitro fermentation by human fecal microbiota. As fermentation progressed, the total carbohydrates of WE were significantly more utilized than that of AE (p < 0.05). The AXs in WE were utilized at a higher rate than those in AE. The relative abundance of Prevotella_9 , which can efficiently utilize AXs, was significantly increased in AE. The presence of AXs in AE shifted the balance away from protein fermentation and caused a delay in protein fermentation. Our study demonstrated that wheat bran CWPs can modulate the gut microbiota in a structure-dependent manner. However, future studies should further characterize the fine structure of wheat CWPs to clarify their detailed relationship with gut microbiota and metabolites. [Display omitted] [ABSTRACT FROM AUTHOR]

Background: Cancer stem-like traits contribute to prostate cancer (PCa) progression and metastasis. Deciphering the novel molecular mechanisms underlying stem-like traits may provide important insight for developing novel therapeutics. Methods: Immunohistochemistry and immunofluorescence assays in prostatic tissues; gain- and loss-of-function analyses using ectopic overexpression and shRNAs in PCa cell lines; measurements of tumorigenic and stemness properties, and transcription in vitro and in vivo; transcriptional analysis in public databases. Results: We identified that overexpression of BTF3 in PCa tissues and BTF3 expression highly correlates to stem-like traits. Cancer stem-like characteristics in PCa including self-renewal and metastatic potential were impaired by BTF3 loss and promoted by BTF3 overexpression. Mechanistically, BTF3 could stabilize BMI1, which is a crucial regulator of prostate stem cell self-renewal. More importantly, our data revealed that BTF3 is highly predictive of poor prognosis and may help in risk stratification of PCa patients. Conclusions: BTF3 promotes PCa progression though modeling stem-like traits in PCa. BTF3 represents a stratification marker in PCa progression and outcomes. [ABSTRACT FROM AUTHOR]

A highly enantioselective organocatalytic Michael addition‐acetalation/oxa‐Michael reaction involving aldehydes and dioxopyrrolidines has been discovered. The asymmetric reaction is mediated by modularly designed organocatalysts (MDOs) self‐assembled from cinchona alkaloid derivatives and amino acids providing a range of optically active tricyclic hexahydro‐2H‐pyrano[3′,2′:5,6]pyrano[2,3‐c]pyrrol derivatives in high yields (up to 99%) with good to excellent enantioselectivities (up to 99% ee) under mild reaction conditions. [ABSTRACT FROM AUTHOR]

Abstract The 2017 Moijabana earthquake in central Botswana (M w 6.4) was a large and deep event for a continental interior and occurred in a region with little historical seismicity. Based on InSAR measurements of surface deformation spanning the event and teleseismic observations, we determine the ruptured fault plane and finite rupture model of the earthquake. Although this oblique normal-faulting earthquake is too deep to uniquely determine the rupture plane geometry from InSAR alone, the best-fitting fault plane constrained by the joint inversion of teleseismic waveforms and InSAR data has a southwest dip and a strike of 126°, roughly consistent with the geologically mapped strike of the Kaapvaal craton's northern edge. Our results indicate that the earthquake had a total duration of ∼10 s, characterized by two major asperities. The first asperity nucleated in the lower crust and then the rupture propagated up-dip. The lower crustal asperity shows a much shorter rise time compared with the shallower asperity, indicating that contrasts in stress or material properties may have played an important role in the rupture process. The earthquake appears to have occurred in the Limpopo belt, a Proterozoic orogenic belt that represents an ancient zone of weakness between the Archean Zimbabwe and Kaapvaal cratons. In the present day, this zone of weakness may be responding to the stress field imposed by the East African Rift System. Highlights • We invert Sentinel-1 InSAR data and teleseismic waveforms for the rupture parameters of the M6.4 Moijabana earthquake in Botswana. • The joint inversion prefers a rupture plane that dips to the southwest with a hypocentral depth of 23 km. • This intraplate earthquake took place in a Proterozoic zone of weakness between two cratonic blocks. [ABSTRACT FROM AUTHOR]

Energy harvesting instead of battery is a better power source for wearable devices due to many advantages such as long operation time without maintenance and comfort to users. However, harvested energy is naturally unstable and program execution will be interrupted frequently. To solve this problem, nonvolatile processor (NVP) has been proposed because it can back up volatile state before the system energy is depleted. However, this backup process also introduces non-negligible energy and area overhead. To improve the performance of NVP, retention state has been proposed recently which can enable a system to retain the volatile data to wait for power resumption instead of saving data immediately. The goal of this paper is to forward program execution as much as possible by exploiting retention state. Specifically, two objectives are achieved. The first objective is to minimize power failures of the system if there is a great probability to get power resumption during retention state. The second objective of this paper is to achieve maximum computation efficiency if it is unlikely to avoid power failure. Compared to the instant backup scheme, evaluation results report that power failure can be reduced by 81.6% and computation efficiency can be increased by 2.5x by the proposed retention state-aware energy management strategy. [ABSTRACT FROM AUTHOR]

Background: Chrysanthemum indicum L., an important ancestral species of the flowering plant chrysanthemum, can be used as medicine and for functional food development. Due to the lack of hereditary information for this species and the difficulty of germplasm identification, we herein provide new genetic insight from the perspective of intraspecific transcriptome comparison and present single sequence repeat (SSR) molecular marker recognition technology. Results: Through the study of a diploid germplasm (DIWNT) and a tetraploid germplasm (DIWT), the following outcome were obtained. (1) A significant difference in Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) annotations for specific homologous genes was observed using the OrthoMCL method for the identification of homologous gene families between the two cytotypes. Ka/Ks analysis of common, single-copy homologous family members also revealed a greater difference among genes that experienced positive selection than among those experiencing positive selection. (2) Of more practical value, 2575 SSR markers were predicted and partly verified. We used TaxonGap as a visual tool to inspect genotype uniqueness and screen for high-performance molecular loci; we recommend four primers of 65 randomly selected primers with a combined identification success rate of 88.6% as priorities for further development of DNA fingerprinting of C. indicum germplasm. Conclusions: The SSR technology based on next-generation sequencing was proved to be successful in the identification of C. indicum germplasms. And the information on the intraspecfic genetic divergence generated by transcriptome comparison deepened the understanding of this complex species' nature. [ABSTRACT FROM AUTHOR]

• Focusing on the effect of clamping pressure on interfacial fusion morphology and mechanical property of CFRTP/Ti alloy laser bonding joint. • Increasing clamping pressure is beneficial to the material diffusion behavior at the joining interface. • Fracture mechanisms of joints under various clamping pressure are discussed based on fusion characteristics and fracture surface morphology. Carbon fiber reinforced thermoplastic composites (CFRTP) and titanium (Ti) alloy are both important lightweight materials in aircraft and aerospace applications, whose heterogeneous hybrid structure manufactured by laser joining technology is propitious to leverage the performance advantages of each component. The clamping pressure during the laser joining procedure is one of the most critical process parameters affecting the joint performance. Laser joining of CFRTP to Ti alloy (Ti6Al4V) under various clamping pressure is carried out, aimed to investigate the effect of clamping pressure on interfacial fusion morphology, mechanical property and fracture mechanism of bonding joint featuring blind hole surface microtextures. The fusion morphology and element distribution at joining interface are observed by scanning electron microscope (SEM) and energy dispersive spectrometer (EDS). Moreover, the fracture morphology and fracture mechanism are analyzed after tensile-shear test. The results show that an appropriate increase of clamping pressure improves the filling effect of melted resin to microtextures on Ti alloy surface, while too high pressure is inclined to induce the generation of pores and cracks at the joining interface. The fracture load of the joints increases and then decreases as the pressure rises, and it reaches a peak value of 3821.4 N corresponding to the optimal interfacial fusion effect when the pressure is 0.8 MPa. Significant distinctions exist in the main fracture mechanisms under different clamping pressure conditions, which lead to different fracture surface morphologies and mechanical performance of laser bonding joints. [ABSTRACT FROM AUTHOR]

Objective To investigate the role of miR-221/222 in cell proliferation and apoptosis in human prostate cancer cells, and examine the effects of miR-221/222 on caspase-10 expression. Methods Prostate cancer cells were transfected with miR-221/222 mimics or inhibitors. Cell proliferation was assessed by MTT assay. The expression levels of miR-221/222 were detected with quantitative real-time PCR. Apoptosis was induced with TNF-α/CHX treatment, and evaluated by Hoechst 33342 staining, propidium iodide (PI) flow cytometric analysis, caspase-3 activity measurement, and Western blot analysis. Luciferase activity assay, quantitative real-time PCR, and Western blot were performed to evaluate the effects of miR-221/222 on caspase-10 expression. Results Our results showed that miR-221/222 could promote the proliferation of prostate cancer cells, including LNCaP and PC3 cells. After transfection and apoptosis induction, Hoechst 33342 staining and PI flow cytometric assay showed that apoptosis was dramatically decreased in prostate cancer cells treated with miR-221/222 mimics. Moreover, caspase-3 activity was dramatically decreased, and the cleaved forms of caspase-3 were reduced, in the miR-221/222 mimic-treated group. On the contrary, miR-221/222 knockdown sensitized the prostate cancer cells to TNF-α/CHX-induced apoptosis. In addition, a negative correlation was observed between the expressions of miR-221/222 and caspase-10 in prostate cancer cells. miR-221/222 could repress the expression of caspase-10, which was confirmed by the luciferase reporter assay. Conclusion miR-221/222 promote cell proliferation and repress apoptosis, through suppressing caspase-10, in prostate cancer cells. Our results provide promising evidence for the miRNA-based therapeutic strategy of prostate cancers. [ABSTRACT FROM AUTHOR]

If a honey bee ( Apis spp.) colony becomes queenless, about 1/3 of young workers activate their ovaries and produce haploid male-producing eggs. In doing so queenless workers maximize their inclusive fitness because the normal option of vicarious production of relatives via their queen’s eggs is no longer available. But if many workers are engaged in reproduction, how does a queenless colony continue to feed its brood and forage? Here we show that in the Asian hive bee Apis cerana hypopharyngeal gland (HPG) size is larger in queenless workers than in queenright workers and that bees undertaking brood-rearing tasks have larger HPG than same-aged bees that are foraging. In queenless colonies, workers with a smaller number of ovarioles are more likely to have activated ovaries. This reinforces the puzzling observation that a large number of ovarioles reduces reproductive success in queenless A. cerana . It further suggests that reproductive workers either avoid foraging or transition to foraging later in life than non-reproductive workers. Finally, our study also showed that ovary activation and larger-than-average numbers of ovarioles had no statistically detectable influence on foraging specialization for pollen or nectar. [ABSTRACT FROM AUTHOR]

Amomum villosum Lour., produced from Yangchun, Guangdong Province, China, is a Daodi medicinal material of Amomi Fructus in traditional Chinese medicine. This herb germplasm should be accurately identified and collected to ensure its quality and safety in medication. In the present study, single nucleotide polymorphism typing method was evaluated on the basis of DNA barcoding markers to identify the germplasm of Amomi Fructus. Genomic DNA was extracted from the leaves of 29 landraces representing three Amomum species (A. villosum Lour., A. xanthioides Wall. ex Baker and A. longiligulare T. L. Wu) by using the CTAB method. Six barcoding markers (ITS, ITS2, LSU D1–D3, matK, rbcL and trnH-psbA) were PCR amplified and sequenced; SNP typing and phylogenetic analysis were performed to differentiate the landraces. Results showed that high-quality bidirectional sequences were acquired for five candidate regions (ITS, ITS2, LSU D1–D3, matK, and rbcL) except trnH-psbA. Three ribosomal regions, namely, ITS, ITS2, and LSU D1–D3, contained more SNP genotypes (STs) than the plastid genes rbcL and matK. In the 29 specimens, 19 STs were detected from the combination of four regions (ITS, LSU D1–D3, rbcL, and matK). Phylogenetic analysis results further revealed two clades. Minimum-spanning tree demonstrated the existence of two main groups: group I was consisting of 9 STs (ST1–8 and ST11) of A. villosum Lour., and group II was composed of 3 STs (ST16–18) of A. longiligulare T.L. Wu. Our results suggested that ITS and LSU D1–D3 should be incorporated with the core barcodes rbcL and matK. The four combined regions could be used as a multiregional DNA barcode to precisely differentiate the Amomi Fructus landraces in different producing areas. [ABSTRACT FROM AUTHOR]

In this paper we describe a rapid, simple, and cost-effective liquid chromatography-tandem mass spectrometric (LC-MS-MS) method for simultaneous analysis of aflatoxin B, B, G, and G, ochratoxin A, and sterigmatocystin in 25 traditional Chinese medicines (TCMs). The method is based on single extraction with 84:16 ( v/ v) acetonitrile-water then analysis of the diluted crude extract without further clean-up. Chromatographic separation was achieved on a C column, with a mobile phase gradient prepared from aqueous 4 mmol L ammonium acetate-0.1 % formic acid and methanol. Quantification of the analytes was by selective reaction monitoring (SRM) on a triple-quadrupole mass spectrometer in positive-ionization mode. Special focus was on investigating and reducing matrix effects to improve accuracy. The established method was validated by determination of linearity ( r > 0.995), sensitivity (limits of quantification 1.6-25.0 ng L), apparent recovery (84.8-110.6 %), extraction recovery (83.6-106.1 %), and precision (relative standard deviation ≤9.9 %) for two representative TCMs, Semen Armeniacae Amarae and Radix Pseudostellariae. The applicability of the method to TCMs other than these was further investigated, and 23 other TCMs with acceptable matrix effects (80.2-118.6 %) were screened. The validated method was finally used to assess mycotoxin contamination of 244 samples of 25 TCMs collected from local hospitals and TCM pharmacies. Aflatoxin B and ochratoxin A were detected in 5.3 % of the samples. Sterigmatocystin, the most prevalent mycotoxin contaminant, was present in 26.2 % of the samples tested; this has not been reported previously. The results of this work imply greater attention should be devoted to evaluation of the potential hazard caused by sterigmatocystin in TCMs. [ABSTRACT FROM AUTHOR]

Abstract: Aims: We aim to investigate the relationship between 1,25-(OH)2VD3 and hsa-let-7a in lung cancer A549 cells. Methods: Real-time PCR and luciferase reporter assays were used to detect the influence of 1,25-(OH)2VD3 on the expression of hsa-let-7a-2 after A549 cells were treated with 1,25-(OH)2VD3 (10−8 ~10−6 mol/L). Analysis of the 5.0Kb upstream sequence of the pre-let-7a-2 showed that one vitamin D response element (VDRE) is located in -2066/-2042bp of pre-let-7a-2. Electrophoretic mobility shift assays (EMSA), chromatin immunoprecipitation (ChIP) and luciferase reporter assays were performed to determine whether 1,25-(OH)2VD3 activating vitamin D receptor (VDR) could bind to this VDRE to promote hsa-let-7a-2 expression. Results: We found that 1,25-(OH)2VD3 could up-regulate the expression of hsa-let-7a-2 in a dose-dependent manner. The results of EMSA and ChIP demonstrated that 1,25-(OH)2VD3/VDR could interact with the VDRE in the upstream of pre-let-7a-2. Luciferase reporter assay showed that this VDRE is a functional cis-element mediating the up-regulation of hsa-let-7a-2 expression induced by 1,25-(OH)2VD3. Conclusions: Our data indicated that 1,25-(OH)2VD3 could up-regulate the transcription of hsa-let-7a-2 in lung cancer cells, and the up-regulation of hsa-let-7a-2 expression induced by 1,25-(OH)2VD3 might mediate the anti-proliferation effects of 1,25-(OH)2VD3 in lung cancer cells. [Copyright &y& Elsevier]

• Patchouli v-myb avian myeloblastosis viral oncogene homolog-related family was analyzed. • PatSWC4 was significantly induced by methyl jasmonate. • PatSWC4 binds to the PatPTS promoter and increases its transcriptional activity. • PatSWC4 interacts with PatJAZ4. • Overexpression of PatSWC4 markedly increased patchoulol production in patchouli. Patchoulol, the major active ingredient in Pogostemon cablin (Blanco) Benth., has considerable clinical and industrial value. A number of genes that participate in the patchoulol biosynthesis pathway have been identified; however, little is known about the transcription factors involved in regulating patchoulol synthesis. In this study, PatSWC4, a homologous protein of Arabidopsis SWC4 belonging to the MYB (v-myb avian myeloblastosis viral oncogene homolog)-related transcription factor family, was identified and characterized. Subcellular localization showed that PatSWC4 protein localized in the nucleus. The highest expression level of PatSWC4 gene was found in old leaves of P. cablin and it was significantly induced by methyl jasmonate (MeJA). Additionally, yeast one-hybrid (Y1H) and dual-luciferase (dual-LUC) assays revealed that PatSWC4 could bind to the promoter of PatPTS gene to increase its transcriptional activity. Yeast two-hybrid (Y2H) assays also confirmed the interaction between PatSWC4 and PatJAZ4 proteins, indicating that PatSWC4 might function in the JA response network. Moreover, transient overexpression of PatSWC4 gene in P. cablin leaves markedly increased the production of patchoulol, and qRT-PCR analysis further revealed that genes in the patchoulol biosynthesis pathway were significantly upregulated at the transcriptional level. Ultimately, we proposed a work model depicting how JA signaling regulates PatPTS via the interaction between PatSWC4 and PatJAZ4, thereby regulating the biosynthesis of patchoulol. Our findings not only help to elucidate the regulatory mechanism governing JA-induced patchoulol biosynthesis, but also lay a foundation for future studies on improving patchoulol production through genetic and metabolic engineering. [ABSTRACT FROM AUTHOR]

Renal fibrosis is the typical manifestation of progressive kidney disease and causes a severe threat to human health. Surging evidence has illustrated that miRNA plays a core role in the genesis and development of kidney fibrosis. MiR-542-3p has been testified to function as a facilitator in hepatic stellate cell activation and fibrosis. The purpose of study is to investigate the potential of miR-542-3p in renal tubulointerstitial fibrosis. In this study, to establish renal fibrosis model in vivo and in vitro , we first conducted unilateral ureteral obstruction (UUO) on rats and high glucose (HG) treatment on the HK-2 cells. Histological and western blot analyses were utilized for assessment of renal fibrosis model. Luciferase reporter assay was carried out to explore the regulatory mechanism underlying miR-542-3p in renal fibrosis. MiR-542-3p was found to be highly expressed in renal fibrosis. Functional experiments revealed that overexpression of miR-542-3p accelerated the deterioration of kidney fibrosis and inhibition of miR-542-3p led to the opposite result. Through the aid of bioinformatics tool, the speculated miR-542-3p binding sites were uncovered in the 3'UTR of argonaute RISC component 1 (AGO1). Mechanism study elucidated that AGO1 was a direct target of miR-542-3p. Lastly, our findings suggested that miR-542-3p played a promoting role in renal fibrosis via repression of AGO1. We justified that miR-542-3p induced kidney fibrogenesis both in vivo and in vitro through targeting AGO1, unveiling that miR-542-3p might be a promising option for the treatment of patients with renal fibrosis. [ABSTRACT FROM AUTHOR]

Amyrins are the immediate precursors of many pharmaceutically important pentacyclic triterpenoids. Although various amyrin synthases have been identified, little is known about the relationship between protein structures and the constituent and content of the products. IaAS1 and IaAS2 identified from Ilex asprella in our previous work belong to multifunctional oxidosqualene cyclases and can produce α-amyrin and β-amyrin at different ratios. More than 80% of total production of IaAS1 is α-amyrin; while IaAS2 mainly produces β-amyrin with a yield of 95%. Here, we present a molecular modeling approach to explore the underlying mechanism for selective synthesis. The structures of IaAS1 and IaAS2 were constructed by homology modeling, and were evaluated by Ramachandran Plot and Verify 3D program. The enzyme-product conformations generated by molecular docking indicated that ASP484 residue plays an important role in the catalytic process; and TRP611 residue of IaAS2 had interaction with β-amyrin through π–σ interaction. MM/GBSA binding free energy calculations and free energy decomposition after 50 ns molecular dynamics simulations were performed. The binding affinity between the main product and corresponding enzyme was higher than that of the by-product. Conserved amino acid residues such as TRP257; TYR259; PHE47; TRP534; TRP612; and TYR728 for IaAS1 (TRP257; TYR259; PHE473; TRP533; TRP611; and TYR727 for IaAS2) had strong interactions with both products. GLN450 and LYS372 had negative contribution to binding affinity between α-amyrin or β-amyrin and IaAS1. LYS372 and ARG261 had strong repulsive effects for the binding of α-amyrin with IaAS2. The importance of Lys372 and TRP612 of IaAS1, and Lys372 and TRP611 of IaAS2, for synthesizing amyrins were confirmed by site-directed mutagenesis. The different patterns of residue–product interactions is the cause for the difference in the yields of two products. [ABSTRACT FROM AUTHOR]

Background: Pogostemon cablin (Blanco) Benth. (Patchouli) is an important aromatic and medicinal plant and widely used in traditional Chinese medicine as well as in the perfume industry. Patchoulol is the primary bioactive component in P. cablin, its biosynthesis has attracted widespread interests. Previous studies have surveyed the putative genes involved in patchoulol biosynthesis using next-generation sequencing method; however, technical limitations generated by short-read sequencing restrict the yield of full-length genes. Additionally, little is known about the expression pattern of genes especially patchoulol biosynthesis related genes in response to methyl jasmonate (MeJA). Our understanding of patchoulol biosynthetic pathway still remained largely incomplete to date. Results: In this study, we analyzed the morphological character and volatile chemical compounds of P. cablin cv. 'Zhanxiang', and 39 volatile chemical components were detected in the patchouli leaf using GC-MS, most of which were sesquiterpenes. Furthermore, high-quality RNA isolated from leaves and stems of P. cablin were used to generate the first full-length transcriptome of P. cablin using PacBio isoform sequencing (Iso-Seq). In total, 9.7 Gb clean data and 82,335 full-length UniTransModels were captured. 102 transcripts were annotated as 16 encoding enzymes involved in patchouli alcohol biosynthesis. Accorded with the uptrend of patchoulol content, the vast majority of genes related to the patchoulol biosynthesis were up-regulated after MeJA treatment, indicating that MeJA led to an increasing synthesis of patchoulol through activating the expression level of genes involved in biosynthesis pathway of patchoulol. Moreover, expression pattern analysis also revealed that transcription factors participated in JA regulation of patchoulol biosynthesis were differentially expressed. Conclusions: The current study comprehensively reported the morphological specificity, volatile chemical compositions and transcriptome characterization of the Chinese-cultivated P. cablin cv. 'Zhanxiang', these results contribute to our better understanding of the physiological and molecular features of patchouli, especially the molecular mechanism of biosynthesis of patchoulol. Our full-length transcriptome data also provides a valuable genetic resource for further studies in patchouli. [ABSTRACT FROM AUTHOR]