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To develop and assess an automated Sub-arc Collimator Angle Optimization (SACAO) algorithm and Cumulative Blocking Index Ratio (CBIR) metrics for single-isocenter coplanar volumetric modulated arc therapy (VMAT) to treat multiple brain metastases. This study included 31 patients with multiple brain metastases, each having 2 to 8 targets. Initially, for each control point, the MLC blocking index was calculated at different collimator angles, resulting in a two-dimensional heatmap. Optimal sub-arc segmentation and collimator angle optimization were achieved using an interval dynamic programming algorithm. Subsequently, VMAT plans were designed using two approaches: SACAO and the conventional Full-Arc Fixed Collimator Angle. CBIR was calculated as the ratio of the cumulative blocking index between the two plan approaches. Finally, dosimetric and planning parameters of both plans were compared. Normal brain tissue, brainstem, and eyes received better protection in the SACAO group (P < 0.05).Query Notable reductions in the SACAO group included 11.47% in gradient index (GI), 15.03% in monitor units (MU), 15.73% in mean control point Jaw area (A Jaw,mean ), and 19.14% in mean control point Jaw-X width (W Jaw-X,mean ), all statistically significant (P < 0.001). Furthermore, CBIR showed a strong negative correlation with the degree of plan improvement. The SACAO method enhanced protection of normal organs while improving transmission efficiency and optimization performance of VMAT. In particular, the CBIR metrics show promise in quantifying the differences specifically in the 'island blocking problem' between SACAO and conventional VMAT, and in guiding the enhanced application of the SACAO algorithm., (© 2024. Australasian College of Physical Scientists and Engineers in Medicine.)

The limited response of hepatocellular carcinoma (HCC) to chemotherapy drugs has always been a bottleneck in therapy. DNA damage repair is a major reason for chemoresistance. Previous studies have confirmed that KIN17 affects chemosensitivity. In this study, we examined the impact of KIN17 on chemotherapy response and DNA repair in HCC cells treated with oxaliplatin (L-OHP). We evaluated the expression and biological roles of KIN17 in HCC using bioinformatic analysis. The correlation between KIN17 and RAD51, particularly their nuclear expression levels, was evaluated using immunofluorescence, immunoblotting after nucleocytoplasmic separation in HCC cells, and immunohistochemistry of mouse xenograft tumors and human HCC tissues. The results indicated a significant increase in KIN17 expression in HCC tissues compared to normal tissues. The GSEA analysis revealed that upregulation of KIN17 was significantly associated with DNA damage repair. Knockdown of KIN17 led to increased DNA damage and reduced cellular survival after exposure to L-OHP. On the other hand, overexpression of KIN17 was linked to decreased DNA damage and improved cell survival following L-OHP treatment. Further experiments indicated that KIN17 affects the expression of RAD51, particularly in the nucleus. KIN17 plays a crucial role in influencing the sensitivity of HCC to chemotherapy by triggering the DNA repair response. Increased expression of KIN17 is associated with a poor prognosis for HCC patients, indicating that KIN17 could serve as a prognostic marker and therapeutic target for HCC., (© 2024. The Author(s) under exclusive licence to Japan Human Cell Society.)

Purpose: The segmentation of the heart and great vessels in CT images of congenital heart disease (CHD) is critical for the clinical assessment of cardiac anomalies and the diagnosis of CHD. However, the diverse types and abnormalities inherent in CHD present significant challenges to comprehensive heart segmentation., Methods: We proposed a novel two-stage segmentation approach, integrating a Convolutional Neural Network (CNN) with a postprocessing method with conditioned energy function for pulmonary and aorta. The initial stage employs a CNN enhanced by a gated self-attention mechanism for the segmentation of five primary heart structures and two major vessels. Subsequently, the second stage utilizes a conditioned energy function specifically tailored to refine the segmentation of the pulmonary artery and aorta, ensuring vascular continuity., Results: Our method was evaluated on a public dataset including 110 3D CT volumes, encompassing 16 CHD variants. Compared to prevailing segmentation techniques (U-Net, V-Net, Unetr, dynUnet), our approach demonstrated improvements of 1.02, 1.04, and 1.41% in Dice Coefficient (DSC), Intersection over Union (IOU), and the 95th percentile Hausdorff Distance (HD95), respectively, for heart structure segmentation. For the two great vessels, the enhancements were 1.05, 1.07, and 1.42% in these metrics., Conclusion: The outcomes on the public dataset affirm the efficacy of our proposed segmentation method. Precise segmentation of the entire heart and great vessels can significantly aid in the diagnosis and treatment of CHD, underscoring the clinical relevance of our findings., (© 2024. CARS.)

Purpose: To improve tumor uptake and prolong tumor retention, a novel fibroblast activation protein (FAP) ligand based on a quinoline-based FAP inhibitor (FAPI) conjugated with the Gly-Pro sequence and 1,4,7,10-tetraazacyclododecane-N,N',N″,N‴-tetraacetic acid (DOTA) was radiolabeled with [ 68 Ga]GaCl 3 ([ 68 Ga]Ga-DOTA-GPFAPI-04). Due to the tumor heterogeneity, this study aimed to further validate the preclinical value of [ 68 Ga]Ga-DOTA-GPFAPI-04 PET imaging in tumor mice models with different FAP expression levels., Methods: [ 68 Ga]Ga-DOTA-GPFAPI-04 was synthesized and its partition coefficient was measured. The stability of [ 68 Ga]Ga-DOTA-GPFAPI-04 was tested in phosphate-buffered saline (PBS, pH 7.4) and fetal bovine serum (FBS). Small animal PET and semi-quantitative studies were conducted in Panc-1 and A549 xenograft tumor mice models compared with [ 68 Ga]Ga-DOTA-FAPI-04. Immunofluorescent and immunohistochemical staining and western blot assay were performed to confirm FAP expression in xenograft tumors., Results: [ 68 Ga]Ga-DOTA-GPFAPI-04 exhibited a radiochemical purity of > 99% and high stability in PBS and FBS. [ 68 Ga]Ga-DOTA-GPFAPI-04 had higher hydrophilic property than [ 68 Ga]Ga-DOTA-FAPI-04 (-4.09 ± 0.05 vs -3.45 ± 0.05). Small animal PET and semi-quantitative analysis revealed Panc-1 xenograft tumor displayed higher tumor uptake of [ 68 Ga]Ga-DOTA-GPFAPI-04 and tumor-to-background ratios compared to A549 xenograft tumor, consistent with the results of immunofluorescence, immunohistochemistry, and western blot. Moreover, [ 68 Ga]Ga-DOTA-GPFAPI-04 demonstrated higher tumor accumulation and longer tumor retention than [ 68 Ga]Ga-DOTA-FAPI-04 in both Panc-1 and A549 xenograft tumors. Furthermore, the FAP-binding specificity of [ 68 Ga]Ga-DOTA-GPFAPI-04 was confirmed in vivo by co-injection of unlabeled GPFAPI-04., Conclusion: [ 68 Ga]Ga-DOTA-GPFAPI-04 showed more favorable in vivo tumor imaging and longer tumor retention compared to [ 68 Ga]Ga-DOTA-FAPI-04, which has high potential to be a promising PET probe for detecting FAP-positive tumors., (© 2024. The Author(s), under exclusive licence to World Molecular Imaging Society.)

Purpose: To investigate the value of shear-wave elastography (SWE) in assessing the response to neoadjuvant chemoradiotherapy (nCRT) in locally advanced rectal cancer., Methods: In this study, 455 participants with locally advanced rectal cancer who underwent nCRT at our hospital between September 2021 and December 2022 were prospectively enrolled. The participants were randomly divided into training and test cohorts in a 3:2 ratio. Clinical baseline data, endorectal ultrasound examination data, and SWE measurements were collected for all participants. Logistic regression models were used to predict whether rectal cancer after nCRT had a low T staging (ypT 0-2 stage, Model A) and pathological complete response (pCR) (Model B). Paired Chi-square tests were used to compare the diagnostic performances of the radiologists to those of Models A and B., Results: In total, 256 participants were included. The area under the receiver operating characteristic curve of Models A and B in the test cohort were 0.94 (0.87, 1.00) and 0.88 (0.80, 0.97), respectively. The optimal diagnostic thresholds for Models A and B were 14.9 kPa for peritumoral mesangial Emean and 15.2 kPa for tumor Emean, respectively. The diagnostic performance of the radiologists was significantly lower than that of Models A and B, respectively (p < 0.05)., Conclusion: SWE can be used as a feasible method to evaluate the treatment response of nCRT for locally advanced rectal cancer., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Management of cardiovascular disease in pregnancy is important, yet the association between cardiovascular health and infertility is rarely reported. In this study, we aimed to explore the association between Life's Essential 8 (LE8), a novel cardiovascular health (CVH) measure, and infertility, and to investigate potential mediating mechanisms. This study investigated cross-sectional data from the 2013-2018 National Health and Nutrition Examination Survey. LE8 score (ranging from 0 to 100) was calculated as the unweighted average of eight CVH metrics. The association between LE8 and infertility was explored through weighted multiple logistic regression. Restricted cubic splines were used to explore nonlinear correlation. In addition, mediation analysis was conducted to investigate the role of oxidative stress and inflammatory markers systematically. After strict exclusion criteria, 1703 American women aged 18-45 years were included. After full adjustment, the LE8 score showed a negative correlation with infertility [per 1 SD increase, OR = 0.675, 95% CI: 0.553-0.824], with a linear dose-response relationship (non-linear P = 0.122). Similar linear negative correlations were found between health factor scores and infertility, with higher body mass index and glucose scores having a significantly lower risk of infertility. Stratified analyses showed a stronger inversed relationship between LE8 and infertility in younger populations. Moreover, mediation analysis revealed that uric acid concentration and lymphocyte count mediated the effect of LE8 on infertility (P < 0.05). LE8 and its subscale scores were linearly and negatively associated with infertility, which may be mediated in part through uric acid and lymphocyte count. Focusing on weight management and glycemic control can effectively reduce the risk of infertility., (© 2024. The Author(s), under exclusive licence to Society for Reproductive Investigation.)

Key Message: The VaBAM3 cloned from Vitis amurensis can enhance the cold tolerance of overexpressed plants, but VaBAM3 knock out by CRISPR/Cas9 system weakened grape callus cold tolerance. In grape production, extreme cold conditions can seriously threaten plant survival and fruit quality. Regulation of starch content by β-amylase (BAM, EC: 3.2.1.2) contributes to cold tolerance in plants. In this study, we cloned the VaBAM3 gene from an extremely cold-tolerant grape, Vitis amurensis, and overexpressed it in tomato and Arabidopsis plants, as well as in grape callus for functional characterization. After exposure to cold stress, leaf wilting in the VaBAM3-overexpressing tomato plants was slightly less pronounced than that in wild-type tomato plants, and these plants were characterized by a significant accumulation of autophagosomes. Additionally, the VaBAM3-overexpressing Arabidopsis plants had a higher freezing tolerance than the wild-type counterparts. Under cold stress conditions, the activities of total amylase, BAM, peroxidase, superoxide dismutase, and catalase in VaBAM3-overexpressing plants were significantly higher than those in the corresponding wild-type plants. Furthermore, sucrose, glucose, and fructose contents in these lines were similarly significantly higher, whereas starch contents were reduced in comparison to the levels in the wild-type plants. Furthermore, we detected high CBF and COR gene expression levels in cold-stressed VaBAM3-overexpressing plants. Compared with those in VaBAM3-overexpressing grape callus, the aforementioned indicators tended to change in the opposite direction in grape callus with silenced VaBAM3. Collectively, our findings indicate that heterologous overexpression of VaBAM3 enhanced cold tolerance of plants by promoting the accumulation of soluble sugars and scavenging of excessive reactive oxygen species. These findings provide a theoretical basis for the cultivation of cold-resistant grape and support creation of germplasm resources for this purpose., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

CRISPR/Cas technology is a powerful tool for genome engineering in Aspergillus oryzae as an industrially important filamentous fungus. Previous study has reported the application of the CRISPR/Cpf1 system based on the Cpf1 (LbCpf1) from Lachnospiraceae bacterium in A. oryzae. However, multiplex gene editing have not been investigated using this system. Here, we presented a new CRISPR/Cpf1 multiplex gene editing system in A. oryzae, which contains the Cpf1 nuclease (FnCpf1) from Francisella tularensis subsp. novicida U112 and CRISPR-RNA expression cassette. The crRNA cassette consisted of direct repeats and guide sequences driven by the A. oryzae U6 promoter and U6 terminator. Using the constructed FnCpf1 gene editing system, the wA and pyrG genes were mutated successfully. Furthermore, simultaneous editing of wA and pyrG genes in A. oryzae was performed using two guide sequences targeting these gene loci in a single crRNA array. This promising CRISPR/Cpf1 genome-editing system provides a powerful tool for genetically engineering A. oryzae., (© 2023. Institute of Microbiology, Academy of Sciences of the Czech Republic, v.v.i.)

Deregulation of circular RNAs (circRNAs) is widely recognized in cancer progression. Our study aims to investigate the role of circ&#95;0020460 in the development of cervical cancer (CC) and its potential mechanism of action. Quantitative real-time polymerase chain reaction (qRT-PCR) and western blot assays were used to detect the expression levels of circ&#95;0020460, miR-485-3p and C-X-C motif chemokine ligand 1 (CXCL1). The roles of circ&#95;0020460 on cell proliferation, cell migration, cell invasion, cell apoptosis, and angiogenesis were investigated using cell counting kit-8 (CCK-8) and Ethynyl deoxyuridine (Edu) assay, wound healing assay, transwell assay, flow cytometry assay, and tube formation assay, respectively. The putative relationship predicted by bioinformatics analysis was verified by dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay. Xenograft models were constructed to explore the role of circ&#95;0020460 in vivo. The expression of circ&#95;0020460 and CXCL1 expression were increased, while miR-485-3p expression was declined in CC tissues and cells. Circ&#95;0020460 knockdown suppressed CC cell proliferation, cell migration, cell invasion, angiogenesis, and promoted cell apoptosis. Circ&#95;0020460 functioned as a miR-485-3p sponge to inhibit miR-485-3p level, and the anti-cancer effects mediated by circ&#95;0020460 knockdown were reversed by miR-485-3p inhibitor. MiR-485-3p bound to CXCL1 3' untranslated region (3'UTR) to degrade CXCL1 expression, and the anti-cancer effects of miR-485-3p restoration were impaired by CXCL1 overexpression. Circ&#95;0020460 downregulation inhibited CC xenograft tumor growth. These results suggest that circ&#95;0020460 promoted the malignant behavior of CC cells by modulating the miR-485-3p/CXCL1 axis., (© 2024. The Author(s).)

Background: Enterococcus faecalis is a common cause of healthcare-associated infections. Its resistance to linezolid, the antibiotic of last resort for vancomycin-resistant enterococci, has become a growing threat in healthcare settings., Methods: We analyzed the data of E. faecalis isolates from 26 medical institutions between 2018 and 2020 and performed univariate and multivariate logistic regression analyses to determine the independent predictors for linezolid-resistant E. faecalis (LREFs). Then, we used the artificial neural network (ANN) and logistic regression (LR) to build a prediction model for linezolid resistance and performed a performance evaluation and comparison., Results: Of 12,089 E. faecalis strains, 755 (6.25%) were resistant to linezolid. Among vancomycin-resistant E. faecalis, the linezolid-resistant rate was 24.44%, higher than that of vancomycin-susceptible E. faecalis (p < 0.0001). Univariate and multivariate regression analyses showed that gender, age, specimen type, length of stay before culture, season, region, GDP (gross domestic product), number of beds, and hospital level were predictors of linezolid resistance. Both the ANN and LR models constructed in the study performed well in predicting linezolid resistance in E. faecalis, with AUCs of 0.754 and 0.741 in the validation set, respectively. However, synthetic minority oversampling technique (SMOTE) did not improve the prediction ability of the models., Conclusion: E. faecalis linezolid-resistant rates varied by specimen site, geographic region, GDP level, facility level, and the number of beds. At the same time, community-acquired E. faecalis with linezolid resistance should be monitored closely. We can use the prediction model to guide clinical medication and take timely prevention and control measures., (© 2023. The Author(s).)

Histological assessment of skeletal muscle slices is very important for the accurate evaluation of weightless muscle atrophy. The accurate identification and segmentation of muscle fiber boundary is an important prerequisite for the evaluation of skeletal muscle fiber atrophy. However, there are many challenges to segment muscle fiber from immunofluorescence images, including the presence of low contrast in fiber boundaries in immunofluorescence images and the influence of background noise. Due to the limitations of traditional convolutional neural network-based segmentation methods in capturing global information, they cannot achieve ideal segmentation results. In this paper, we propose a muscle fiber segmentation network (MF-Net) method for effective segmentation of macaque muscle fibers in immunofluorescence images. The network adopts a dual encoder branch composed of convolutional neural networks and transformer to effectively capture local and global feature information in the immunofluorescence image, highlight foreground features, and suppress irrelevant background noise. In addition, a low-level feature decoder module is proposed to capture more global context information by combining different image scales to supplement the missing detail pixels. In this study, a comprehensive experiment was carried out on the immunofluorescence datasets of six macaques' weightlessness models and compared with the state-of-the-art deep learning model. It is proved from five segmentation indices that the proposed automatic segmentation method can be accurately and effectively applied to muscle fiber segmentation in shank immunofluorescence images., (© 2023. The Author(s) under exclusive licence to Society for Imaging Informatics in Medicine.)

The spent carbon cathode (SCC) is a hazardous solid waste from aluminum production. It has an abundant carbon source and a unique graphitic carbon layer structure, making it a valuable waste for recycling. This paper uses alkaline and acid leaching methods to report a straightforward way of extracting recovered carbon (RC) from SCC as anode material for lithium-ion batteries (LIBs). The results show that alkaline and acid leaching conditions at 70 °C with 1 M NaOH and HCl solution individually in 6 h and a liquid-solid ratio of 20:1 can result in RC with up to 94.63% carbon content than 49.38% in SCC, exhibiting a typical graphite structure. SCC and RC materials are obtained after calcination at 400 °C in an inert atmosphere and used as anode materials (SCC-400 and RC-400). In this paper, The initial charging specific capacities are 490.0 mA h g -1 , 195.4 mA h g -1 , and 423.2 mA h g -1 and initial coulombic efficiencies (ICE) are 67.8%, 78.9%, and 72.0% of RC-400, SCC, and SCC-400. RC-400 also shows excellent capacity retention and impedance values. This exciting finding provides a viable, non-hazardous, and resourceful method for treating and disposing of SCC from aluminum electrolysis., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Phosphogypsum (PG), which has great environmental harm and restricts the sustainable development of phosphorus chemical industry, is a solid waste produced in phosphoric acid production. Thermal decomposition of PG is an extensive way to reutilization of resource, and the key point is to establish an appropriate decomposition path and catalyst system of PG. In the work, the strategy for low-temperature and highly-efficient decomposition of PG is established based on the thermodynamic analysis and the experimental research by metal ions to reduce the decomposition temperature. Meanwhile, SEM(Scanning Electron Microscope) is used to characterize the composition and morphology of PG in the various conditions, also the decomposition temperature is analyzed by TGA(Thermogravimetric Analysis). Then, the decomposition ratio via Fe 3+ /Co 2+ /Ni 2+ unitary/ternary catalyst is obtained by precipitation method. Through kinetic analysis combined with XRF(X-ray Fluorescence Spectrometer) and EDX(Energy Dispersive X-Ray Spectroscopy) results, it is found that there is a reaction competition in the decomposition process by Fe 3+ /Co 2+ /Ni 2+ ternary catalyst. Further the mechanism of catalytic system on PG is derived. The present work can be concluded that Fe 3+ /Co 2+ /Ni 2+ can effectively reduce the decomposition temperature of PG, and the effect of ternary metal is more obvious than that of unitary metal. Finally, pomelo peel is used instead of coke to successfully decompose PG at low temperature by one step method. The establishment of low temperature decomposition system of PG has potential application in phosphorus chemical industry and is in line with sustainable development., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Background: Accessing professional medical interpreters for brief, low risk exchanges can be challenging. Machine translation (MT) for verbal communication has the potential to be a useful clinical tool, but few evaluations exist., Objective: We evaluated the quality of three MT applications for English-Spanish and English-Mandarin two-way interpretation of low complexity brief clinical communication compared with human interpretation., Design: Audio-taped phrases were interpreted via human and 3 MT applications. Bilingual assessors evaluated the quality of MT interpretation on four assessment categories (accuracy, fluency, meaning, and clinical risk) using 5-point Likert scales. We used a non-inferiority design with 15% inferiority margin to evaluate the quality of three MT applications with professional medical interpreters serving as gold standards., Main Measures: Proportion of interpretation exchanges deemed acceptable, defined as a composite score of 16 or greater out of 20 based on the four assessment categories., Key Results: For English to Spanish, the proportion of MT-interpreted phrases scored as acceptable ranged from 0.68 to 0.84, while for English to Mandarin, the range was from 0.62 to 0.76. Both Spanish/Mandarin to English MT interpretation had low acceptable scores (range 0.36 to 0.41). No MT interpretation met the non-inferiority threshold., Conclusion: While MT interpretation was better for English to Spanish or Mandarin than the reverse, the overall quality of MT interpretation was poor for two-way clinical communication. Clinicians should advocate for easier access to professional interpretation in all clinical spaces and defer use of MT until these applications improve., (© 2023. The Author(s), under exclusive licence to Society of General Internal Medicine.)

Allergen-specific immunotherapy (SIT) is a desirable way of therapy for various allergic diseases such as food allergy (FA). However, frequent visits for more than 3 years and potential adverse effects often hinder patient compliance. Recently, many researchers started focusing on microneedles (MNs) as a new method for SIT. In this study, we proposed an implantable MNs system produced by a two-step casting process, consisting of OVA (antigen)-loaded silk microneedles and a dissolvable, flexible polyvinyl alcohol (PVA) pedestal. Different from PVA, silk fibroin hydrogel has preferable vaccine release ability in vivo and in vitro. Once MNs are inserted into the skin, the PVA pedestal can dissolve in the interstitial fluid of the excised skin within 5 min and implant the OVA-loaded silk microneedle tips in dermal layer as a sustained antigen depot, thus inducing long-lasting immune response for at least 2 weeks. After receiving 3 doses of MN-based immunotherapy, the immune response in OVA-sensitized mice was successfully suppressed, with no apparent side effects. Compared to conventional subcutaneous immunotherapy (total dose of 150 [Formula: see text]g), MN immunotherapy ameliorated systemic anaphylaxis more effectively even at a lower dose (total dose of 30 [Formula: see text]g), demonstrating the antigen dose-sparing potential of the proposed MNs. Moreover, due to the prolonged release effect of silk-PVA composite MNs, the frequency of immunotherapy can be significantly reduced. To sum up, through prolonged skin exposure to antigen, this implantable designed MN may offer a new therapeutic strategy for FA treatment with significant improvements in efficacy and convenience. Schematic illustration of silk-PVA composite microneedles, consisting of OVA (antigen)-loaded silk microneedles and a dissolvable, flexible PVA pedestal. Once inserted into the skin, the PVA pedestal can dissolve in the interstitial fluid of the excised skin within 5 min. Subsequently, the OVA-loaded silk microneedle tips were implanted in the dermal layer as a sustained antigen depot and induced long-lasting immune response. This MNs-based immunotherapy can significantly modulate the Th1/Th2 imbalance of sensitized mice., (© 2023. Controlled Release Society.)

Many bioactive secondary metabolites synthesized by fungi have important applications in many fields, such as agriculture, food, medical and others. The biosynthesis of secondary metabolites is a complex process involving a variety of enzymes and transcription factors, which are regulated at different levels. In this review, we describe our current understanding on molecular regulation of fungal secondary metabolite biosynthesis, such as environmental signal regulation, transcriptional regulation and epigenetic regulation. The effects of transcription factors on the secondary metabolites produced by fungi were mainly introduced. It was also discussed that new secondary metabolites could be found in fungi and the production of secondary metabolites could be improved. We also highlight the importance of understanding the molecular regulation mechanisms to activate silent secondary metabolites and uncover their physiological and ecological functions. By comprehensively understanding the regulatory mechanisms involved in secondary metabolite biosynthesis, we can develop strategies to improve the production of these compounds and maximize their potential benefits., (© 2023. The Author(s), under exclusive licence to Springer Nature B.V.)

In order to assess the metal pollution in the Wujiang River, concentrations of Cu, Zn, Fe, Mn, Pb, Cd, As, and Hg in the water, sediments, and two fish species Cyprinus carpio and Pelteobagrus fulvidraco from the middle reaches (Tuomugang, TMG) and lower reaches (Wulong, WL) of the Wujiang River were examined. The results indicated that all metal concentrations were lower than the values for grade one water quality according to the Environment Quality Standard for Surface Water of China (GB 3838-2002). The bioavailable fraction concentrations of Zn, Fe, Mn, Pb, and Hg in WL were significantly higher than those in TMG (p < 0.05), indicating that these metals in sediments of WL have higher bioavailability and mobility. The Cu, Zn, Fe, Pb, As, and Hg were mainly related to the residual fraction, while the Mn and Cd were mainly associated with the non-residual fraction. The risk assessment code (RAC) and the secondary phase to the primary phase (RSP) values indicated that Mn and Cd have a high risk of secondary release. The mean metal concentrations in the liver of the two fish species were higher than those in muscle. The higher metal concentrations of fish in WL suggested that bioaccumulation of metals in fish could be influenced by metal bioavailability. No identical relationships between metal concentrations and fish length were manifested in the present study. The values of target hazard quotient (THQ) and hazard index (HI), and carcinogenic risk (CR) of metals for the consumption of C. carpio and P. fulvidraco indicated that the anglers would likely not experience significant non-carcinogenic risk, but the carcinogenic risk of As cannot be ignored. Thanks to prohibited commercial fishing in the Wujiang River, the metal pollution will probably not pose a health risk to the general public for wild fish consumption., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

This paper explores the effect mechanism of population aging on carbon dioxide emissions using provincial panel data in China from 2005 to 2019 and examines how aging affects carbon dioxide emissions through stepwise regression test and Sobel test respectively by constructing a multiple mediating effect model. The following conclusions have been drawn: (1) the results of the baseline regression show that population aging has a significant inhibitory effect on carbon emissions. (2) The results of intermediary effect test show that the intermediary effect of population aging through technological progress, industrial structure, consumption level, and consumption structure is significant. (3) The results of heterogeneity test show that the mediating effect of technological progress is not significant in the eastern region, central region, and western region, and the mediating effect of industrial structure and consumption structure is significant in the eastern region and central region, and the mediating effect of consumption level is significant in the eastern region and western region. (4) The results of the robustness test also verify the mediating effect conclusion once again and prove the robustness of the results. Overall, as an aging society country, China should take the impact of aging into consideration more in the process of achieving emission peak and carbon neutrality., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Nitrogen oxide (NOx) contains two harmful air pollutants: nitric oxide (NO) and nitrogen dioxide (NO 2 ). The reasonable prediction of China's NOx emissions is of positive significance for the government to formulate environmental protection policies. To this end, a new grey prediction model with second-order differential equation is proposed in this paper, which has more reasonable model structure and better modeling performance than the traditional grey model. Secondly, according to the data characteristics of NOx emissions of China in recent years, a smoothing algorithm and weakening buffer operator are employed to process the original data to solve the rationality of the prediction results of the new model. Thirdly, the model for predicting China's NOx emissions has been constructed by the new proposed model. The results show that the mean comprehensive error of the new model is only 0.0692%, and its performance is much better than that of several other mainstream grey prediction models. Finally, the new model is applied to China's carbon dioxide prediction in the next 5 years, and the rationality of the prediction results is analyzed. Based on the prediction results, relevant countermeasures and suggestions are put forward., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Although 5-methylcytosine (m 5 C) has been identified as a novel and abundant mRNA modification and associated with energy metabolism, its regulation function in adipose tissue and skeletal muscle is still limited. This study aimed at investigating the effect of mRNA m 5 C on adipogenesis and myogenesis using Jinhua pigs (J), Yorkshire pigs (Y) and their hybrids Yorkshire-Jinhua pigs (YJ). We found that Y grow faster than J and YJ, while fatness-related characteristics observed in Y were lower than those of J and YJ. Besides, total mRNA m 5 C levels and expression rates of NSUN2 were higher both in backfat layer (BL) and longissimus dorsi muscle (LDM) of Y compared to J and YJ, suggesting that higher mRNA m 5 C levels positively correlate with lower fat and higher muscle mass. RNA bisulfite sequencing profiling of m 5 C revealed tissue-specific and dynamic features in pigs. Functionally, hyper-methylated m 5 C-containing genes were enriched in pathways linked to impaired adipogenesis and enhanced myogenesis. In in vitro, m 5 C inhibited lipid accumulation and promoted myogenic differentiation. Furthermore, YBX2 and SMO were identified as m 5 C targets. Mechanistically, YBX2 and SMO mRNAs with m 5 C modification were recognized and exported into the cytoplasm from the nucleus by ALYREF, thus leading to increased YBX2 and SMO protein expression and thereby inhibiting adipogenesis and promoting myogenesis, respectively. Our work uncovered the critical role of mRNA m 5 C in regulating adipogenesis and myogenesis via ALYREF-m 5 C-YBX2 and ALYREF-m 5 C-SMO manners, providing a potential therapeutic target in the prevention and treatment of obesity, skeletal muscle dysfunction and metabolic disorder diseases., (© 2022. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

The kojic acid gene cluster of Aspergillus oryzae plays a key role in kojic acid synthesis. Although the kojic acid gene cluster has been found in 2010, there is little information on the function of the genes located near the kojic acid gene cluster of A. oryzae and whether these genes affect the kojic acid gene cluster containing kojA, kojR and kojT. Here, Aokap6 near the kojic acid gene cluster of A. oryzae was identified and characterized. The Aokap6 disrupted mutants were constructed by the CRISPR/Cas9 system, which exhibited increased mycelium growth and conidial formation. Disruption of Aokap6 enhanced the tolerance to cell wall, oxidative and heat stress but not osmotic stress. Deletion of Aokap6 repressed kojic acid production, together with the reduced expression of kojA, kojR and kojT. Meanwhile, knockout of kojA, kojR and kojT led to the declined expression of Aokap6, indicating that Aokap6 is required for kojic acid production in coordination with kojA, kojR and kojT. Furthermore, overexpression of kojA, kojR and kojT had no effects on the transcript level of Aokap6, and overexpression of kojA in Aokap6 deletion strain could rescue the reduced yield of kojic acid, suggesting that Aokap6 is involved in kojic acid synthesis acting upstream of kojA. These findings provide new insight for the further understanding of kojic acid gene cluster and kojic acid production in A. oryzae., (© 2022. The Author(s), under exclusive licence to Springer Nature B.V.)

Cancer has been a serious threat and impact on the health and life of human. Phototherapy is considered as a promising therapeutic method to replace the traditional treatment in clinic owing to its noninvasive nature and high efficiency. Photoinitiators have long been used in the field of photopolymerization; however, few studies have been carried out on their potential as anticancer agents under light irradiation. In this study, the effect of a photoinitiator, diphenyl (2, 4, 6-trimethylbenzoyl) phosphine oxide (TPO), on breast cancer is investigated and the related mechanism is elucidated. It is found that TPO has low dark toxicity and significant phototoxicity. TPO can inhibit cell growth and development and promote cell apoptosis through a mitochondrial pathway under light irradiation. Further studies show that cell apoptosis is induced by free radicals produced from the photolysis of TPO to activate JNK phosphorylation. Overall, we identify the antitumor effects of TPO in vitro for the first time, and provides a proof of concept for its application as a novel photolatent therapeutic drug., (© 2022. The Author(s).)

Kojic acid is an industrially important secondary metabolite produced by Aspergillus oryzae. The construction of genetic materials for kojic acid related genes is important for understanding the mechanism of kojic acid synthesis in A. oryzae. However, multigene simultaneous knockout mutants for kojic acid synthesis genes remain limited because A. oryzae is multinuclear and good selectable markers are scarce. Here, we firstly successfully obtained single mutants of kojA, kojR, and kojT by our previously constructed CRISPR/Cas9 system in A. oryzae, which demonstrated the feasibility of the targeting sgRNAs for kojA, kojR, and kojT. Then, the AMA1-based genome-editing system for multiplex gene editing was developed in A. oryzae. In the multiplex gene-editing system, two guide RNA expression cassettes were ligated in tandem and driven by two U6 promoters in the AMA1-based autonomously replicating plasmid with the Cas9-expression cassette. Moreover, the multiplex gene-editing technique was applied to target the kojic acid synthesis genes kojA, kojR, and kojT, and the double and triple mutants within kojA, kojR, and kojT were obtained successfully. Additionally, the selectable marker pyrG was knocked out in the single and triple mutants of kojA, kojR, and kojT to obtain the auxotrophic strains, which can facilitate to introduce a target gene into the single and triple mutants of kojA, kojR, and kojT for investigating their relationship. The multiplex gene-editing system and release of these materials provide a foundation for further kojic acid research and utilization., (© 2022. Institute of Microbiology, Academy of Sciences of the Czech Republic, v.v.i.)

Being one of the most common wood in furniture and decoration, Fraxinus mandshurica Rupr. (F. mandshurica) is a frequently used material for wood products. However, the resulting odor problem has not yet been solved so far. Thus, this study focused on exploring odor-active compounds and investigating the effect of environmental conditions on emission and odor attributes. F. mandshurica was studied via TD-GC-MS/O combined with microchamber. Totally, 15 kinds of key odor-active compounds were detected from F. mandshurica. The main odorous components were found to be aromatics, aldehydes and ketones, esters, alcohols and acids. Six kinds of attributes played a key role in the overall odor character of F. mandshurica. Fragrant was the predominant odor impressions, followed by vinegar and antiseptic solution. The proportion of the odor-active compounds concentration to total concentration decreased with an increase in the temperature and relative humidity, while the ratio of air exchange rate to the loading factor had little effect on this. The total concentration of F. mandshurica increased as the temperature and relative humidity increased, but these decreased as the ratio of air exchange rate to the loading factor increased, while the total odor intensity reduced with the rise in the ratio of air exchange rate to the loading factor. Time was found to be the most effective factor for eliminating emissions. Considering the environmental conditions in this study, 40 °C, 60% relative humidity and a 1.0 m 3 ·m -2  h -1 ratio of air exchange rate to loading factor are recommended to accelerate the emission of F. mandshurica. The optimum condition was found to be higher temperature, higher relative humidity and higher ratio of air exchange rate to loading factor., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

γ-aminobutyric acid (GABA) has various physiological functions and is widely used in medicine, food, and other fields. Glutamate decarboxylase (GAD) is a key enzyme that catalyzes the decarboxylation of L-glutamate to synthesize GABA. However, the industrial application of microorganism-derived GAD is limited by its rapid loss of enzymatic activity with pH approaching neutrality. In this study, a novel glutamate decarboxylase, GAD MSM, from Mycobacterium smegmatis was overexpressed and purified. On the basis of homologous modeling and substrate molecular docking, several GAD MSM mutants were constructed, and their enzymatic properties were analyzed. The results showed that the optimal pH of wild-type GAD MSM is 5.4; at pH 6.2, 22.8% enzymatic activity was retained. The T211I replacement in GAD and C-terminal deletion mutant GAD MSMΔC showed relatively high catalytic activity in a pH range of 5.0-7.0. The V max and K m values of GAD MSMΔC were 14.69 and 5.70, respectively, at pH 5.5, and 9.87 and 6.17, respectively, at pH 7.0. Compared with the wild-type GAD, GAD MSMΔC maintained higher affinity and enzymatic activity of the substrate, maintaining 78.5% of the highest enzymatic activity even at pH 7.0, which is the highest reported activity retention for GAD under neutral pH condition. Therefore, GAD MSMΔC can be used for the transformation of high-yielding strains and industrial production of GABA., (© 2022. The Author(s), under exclusive licence to Springer Nature B.V.)

This study aimed to examine the protective effects of catalpa on ultrastructure of hippocampal neuron and limb motor function in rats with cerebral ischemia. 90 healthy Sprague-Dawley male rats were randomly divided into control (n = 30) and model (n = 60) groups. Cerebral ischemia and hippocampal neurons were induced by occluding the internal carotid artery and injection of high blood glucose, respectively. Model rats were randomly divided into routine (n = 30) and observational (n = 30) groups. Animals in the routine group received edaravone injection (7 mg/kg/day) for 14 days, while rats in the observation group were treated with catalpol (30 mg/kg/day) for 14 days. Limb motor function score, fine motion execution capability, number of hippocampal neurons retained, and the ultrastructure of hippocampal nerve cells were considered at 3, 7, and 14 days after treatments. A significant difference was observed in the mean scores of limb motor function, fine motor execution ability, and the number of hippocampal neurons retained between groups (p < 0.001). Repetitive treatments with catalpol significantly improved the mean number of hippocampal neurons retained (p < 0.01), limb motor function (p < 0.001), and fine motor execution ability scores (p < 0.01) at 3, 7, and 14 days compared to edaravone. Catalpol treatments also improved the ultrastructure morphology of neuronal cells. Catalpa can effectively improve limb motor function and protect hippocampal neuron function in rats with cerebral ischemia., (© 2021. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Global climate change is a major threat to biodiversity, which may increase the extinction risk of rare species, particularly those like Ostrya rehderiana Chun (Betulaceae) with very few remaining extant wild individuals. We aimed to estimate the potential distribution of O. rehderiana under climate change and to analyze possible relevant climatic factors. Maximum entropy (Maxent) was employed to model the potential distribution of O. rehderiana under present and future climate scenarios. Suitable habitat areas in different periods and the main contributing climate factors were identified using species distribution models. The minimum temperature in winter and precipitation seasonality were the principal climatic factors influencing the establishment of O. rehderiana. The proportion of high potential distribution area in China was 3.91% and would further shrink significantly under changing climate, especially reduce by 97% under high radiative forcing. The extinction risk of O. rehderiana would still be extraordinarily high under future climate scenarios. The Tianmu and Luoxiao Mountains would be the only potential refugia for O. rehderiana in the future. Special conservation efforts are urgently required to rescue extremely endangered species as O. rehderiana. We propose priorities for the conservation region and suggestions for conservation management strategies., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Vermicompost was used as an additive in swine manure composting to investigate the expression of bacterial functional genes on nutrients biotransformation. Three treatments with vermicompost compositions of 10%, 20%, and 30% in swine manure were set up. Raw manure was used as the control. The thermophilic period increased to 12 days, the NH 4 + -N/NO 3 - -N ratio decreased to 0.85, and the germination index (GI) increased to 166% after vermicompost addition. Furthermore, higher relative abundances of Firmicutes were observed in the substrate during the initial stages of experiment. The abundance of the dominant phylum Proteobacteria and its related pathogenic genera Acinetobacter and Stenotrophomonas decreased in the thermophilic stage while the potentially beneficial genera Actinomadura and Chryseolinea increased. The expression of primary functional genes associated with the metabolism of carbohydrates, amino acids, xenobiotics, and fatty acids was enhanced during the thermophilic phase. Besides, most dominant genera showed strengthened correlations with NO 3 - -N and GI, which were the strongest environmental factors for bacterial communities. Network analysis revealed a new metabolic pathway associated with dominant genera Pseudomonas, Acinetobacter, Stenotrophomonas, and Oceanobacter, whose abundance increased with vermicompost addition. Collectively, the results of this study indicate that vermicompost can promote composting efficiency by increasing the potentially beneficial bacteria, decreasing pathogenic bacteria, and enhancing the metabolic capacity of bacterial communities., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

The Nramp (natural resistance-associated macrophage protein) family of genes has been identified and characterized widely in many species. However, the Nramp genes and their characterizations have not been reported for Aspergillus oryzae . Here, only one Nramp gene AoNramp1 in A. oryzae genome was identified. Phylogenetic analysis revealed that AoNramp1 is not clustered with Nramps from yeast genus. Expression analysis showed that the transcript level of AoNramp1 was strongly induced under both Zn/Mn-replete and -deplete conditions. The GUS-staining assay indicated that the expression of AoNramp1 was strongly induced by Zn/Mn. Moreover, the AoNramp1 deletion and overexpression strains were constructed by the CRISPR/Cas9 system and A. oryzae amyB promoter, respectively. Phenotypic analysis showed that overexpression and deletion of AoNramp1 caused growth defects under Zn/Mn-deplete and -replete conditions, including mycelium growth and conidia formation. Together, these findings provide valuable information for further study on the biological roles of AoNramp1 in A. oryzae ., Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-021-02998-z., Competing Interests: Conflict of interestsThe authors declare that they have no conflict of interest., (© King Abdulaziz City for Science and Technology 2021.)

A reasonable prediction of the finished products from waste recycling and reprocessing is of great significance to the sustainable use of limited resources, the reduced pollution caused by waste, and the reflected comprehensive waste utilization (CWU) creativity. To this end, an improved multi-variable grey model was employed to forecast the finished products in China's CWU. Firstly, the degree of grey incidence was applied to select explanatory variables and eliminate multicollinearity between them. Then, compared with the traditional GM(1,N) model, the linear correction term and grey action quantity were added in the proposed improved model, and the response function and parameter estimation method of the improved model were deduced and proved. Thirdly, the finished products from CWU was simulated and predicted by the proposed model. The mean relative simulation percentage error of the improved model was only 0.0001%, in comparison with the ones obtained from the traditional GM(1,N) and the classical GM(1,1), which were 12.1232 and 8.8402%, respectively. Lastly, the results show that the finished products from CWU are mainly affected by the comprehensive utilization of general industrial solid waste and the number of industrial enterprises in CWU, and the future trends from 2020 to 2025 are unstable., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

D-amino acid oxidase activator (DAOA) gene, which plays a crucial role in the process of glutamatergic transmission and mitochondrial function, is frequently linked with the liability for schizophrenia. We aimed to investigate whether the variation of DAOA rs2391191 is associated with alterations in white matter integrity of first-episode schizophrenia (FES) patients; and whether it influences the association between white matter integrity, cognitive function and clinical symptoms of schizophrenia. Forty-six patients with FES and forty-nine healthy controls underwent DTI and were genotyped for DAOA rs2391191. Psychopathological assessments were performed by Brief Psychiatric Rating Scale (BPRS) and Scale for Assessment of Negative Symptoms (SANS). Cognitive function was assessed by MATRICS Consensus Cognitive Battery (MCCB). Schizophrenia patients presented lower fractional anisotropy (FA) and higher radial diffusivity (RD), mainly spreading over the corpus callosum and corona radiata compared with healthy controls. Compared with patients carrying G allele, patients with AA showed lower FA in the body of corpus callosum, and higher RD in the genu of corpus callosum, right superior and anterior corona radiata, and left posterior corona radiata. In patients carrying G allele, FA in body of corpus callosum was positively correlated with working memory, RD in genu of corpus callosum was negatively associated with the speed of processing, working memory, and the composite score of MCCB, while no significant correlations were found in AA homozygotes. In our study, patients with FES presented abnormal white matter integrity in corpus callosum and corona radiata. Furthermore, this abnormality was associated with the genetic variation of DAOA rs2391191, with AA homozygotes showing less white matter integrity in the corpus callosum. Our findings possibly provide further support to the evidence that DAOA regulates the process of glutamatergic neurotransmission and mitochondrial function in the pathophysiological mechanism of schizophrenia., (© 2020. Springer Science+Business Media, LLC, part of Springer Nature.)

The CRISPR/Cas9 system has become a great tool for target gene knock-out in filamentous fungi. It is laborious and time-consuming that identification mutants from a large number of transformants through PCR or enzyme-cut method. Here, we first developed a CRISPR/Cas9 system in Aspergillus oryzae using AMA1-based autonomously replicating plasmid and Cas9 under the control of the Aspergillus nidulans gpdA promoter. By the genome editing technique, we successfully obtained mutations within each target gene in Aspergillus oryzae. Then, we put the protospacer sequence of a target gene and its protospacer adjacent motif (PAM) behind the start codon "ATG" of DsRed, yielding the non‑functional DsRed (nDsRed) reporter gene, and the nDsRed reporter gene could be rescued after successful targeted editing. Moreover, this method was also applied by targeting the kojic acid synthesis gene kojA, and the transformants with DsRed activity were found to harbor targeted mutations in kojA. These results suggest that the nDsRed can be used as a powerful tool to facilitate the identification of mutants generated by CRISPR/Cas9 in Aspergillus oryzae.

Background and Aim: The aim of this study was to evaluate the accuracy of serum markers of liver fibrosis for predicting progression to acute-on-chronic liver failure (ACLF) in patients with acute exacerbation (AE) and severe AE (SAE) of chronic hepatitis B virus (HBV) infection., Methods: The predictive accuracy of aminotransferase-to-platelet ratio index (APRI), Fibrosis-4, Lok index, and Forns index for progression to ACLF was evaluated via receiver operating characteristic (ROC) curve and area under the ROC (AUROC) in 441 and 130 patients with AE and SAE., Results: After admission, 24 (5.8%) and 25 (19.2%) patients with AE and SAE, respectively, progressed to ACLF. The Lok index was one of the independent risk factors associated with progression to ACLF in patients with AE and SAE. The AUROC of Lok index for diagnosing liver cirrhosis was 0.815 (0.774-0.851) in patients with AE and 0.715 (0.629-0.791) in patients with SAE. The AUROC of Lok index for predicting progression to ACLF in patients with AE and SAE was 0.756 (0.711-0.797) and 0.866 (0.795-0.919), respectively. In patients with AE and SAE, the cut-off values of the Lok index for predicting ACLF were higher and lower, respectively, than those for diagnosing liver cirrhosis., Conclusion: The Lok index has predictive accuracy regarding progression to ACLF in patients with AE and SAE. Different thresholds of liver fibrosis are needed for determining progression to ACLF in patients with different severity of liver injury during acute exacerbation of chronic HBV infection., (© 2021. Asian Pacific Association for the Study of the Liver.)

Rab-like 3 (RABL3) is a member of Rab family that is related with several kinds of cancers. However, the functional roles of RABL3 in oral squamous cell carcinoma (OSCC) remain largely unknown. In the current study, we examined the expression levels of RABL3 in OSCC tissues and cell lines. The results showed that RABL3 expression was markedly increased in OSCC tissues and cell lines. Knockdown of RABL3 significantly suppressed the proliferation, migration and invasion of OSCC cells. Overexpression of RABL3 exhibited opposite effects with RABL3 knockdown. In vivo assay demonstrated that knockdown of RABL3 suppressed the tumorigenesis of OSCC. Moreover, RABL3 regulated the activation of focal adhesion kinase (FAK)/protein kinase B (Akt) signaling pathway in OSCC cells. Inhibition of FAK reversed the effects of RABL3 overexpression on cell proliferation, migration and invasion of OSCC cells. In conclusion, these findings demonstrated that RABL3 acted as an oncogene in OSCC, which was attributed to the regulation of FAK/Akt pathway. Thus, RABL3 may be potential therapeutic target for the treatment of OSCC.

Secondary pollution resulting from shoot death is a difficult problem that complicated the application of wetland plants for water purification in northern wetlands. Phalaris arundinacea, a perennial herb with an obviously declining stage, or senescence, is a species that is often selected for water purification in Northern China; however, whether it reduces the secondary pollution risk via nitrogen (N) and phosphorus (P) accumulation during senescence or not remains unclear. To investigate this question, an experiment was conducted with containerized plants during the winter of 2016, after roughly half the leaves on the plants had withered. The experimental observations and analyses were conducted within 0, 2, 4, 6, and 8 weeks of the initiation of senescence. Results revealed that leaves continued to wither and shoot death occurred during weeks 4 to 6 and 8 to 10, respectively. However, no significant differences occurred in fresh biomass or in N and P accumulations of a single plant during senescence. The root biomass, root weight per volume, and total N content increased significantly, while total P content remained stable when leaves withered, respectively. H + -ATPase, a key enzyme for ion transportation, decreased after the leaves withered. However, root activity, evaluated by absorption surface per root volume, remained stable, and percentage of fine root length (diameter < 1 mm) increased significantly during senescence. In conclusion, the root activity and morphology enables P. arundinacea to accumulate N and P during senescence, which makes it a good choice for water purification in northern wetlands.

Glycerol dehydrogenase has been identified and characterized functionally in many species. However, little is known about glycerol dehydrogenase genes and their functions in Aspergillus oryzae. Here, a total of 45 glycerol dehydrogenase genes in Aspergillus oryzae were identified and renamed from AoGld1 to AoGld45 according to their chromosome distribution. They were classified into three groups based on phylogenetic analysis. Synteny analysis revealed that thirteen AoGld genes are conserved among Aspergillus species. Promoter analysis displayed that AoGld3 and AoGld13 harbored multiple binding elements of GATA-type transcription factors and zinc-finger protein msnA that were involved in nitrogen and kojic acid metabolism, respectively. Moreover, the AoGld3 deletion strain Δgld3 was generated by the CRISPR/Cas9 system, which had no visible growth defects compared with the control wild-type strain under the control and osmotic stress treatments. However, disruption of AoGld3 led to the inhibition of kojic acid production, and the expression of kojA, kojR was down-regulated in the Δgld3 strain. Furthermore, when kojA or kojR was overexpressed in the Δgld3 strain, the yield of kojic acid was restored, suggesting that AoGld3 is involved in kojic acid production through affecting the expression of kojR and kojA. Taken together, these findings provide new insights into our understanding of glycerol dehydrogenase and establish foundation for further study of their roles in Aspergillus oryzae.

Membrane proteins are the major component of plasma membranes, and they play crucial roles in all organisms. To understand the influence of the presence of KcsA channel on cell membrane electroporation induced by picosecond pulse trains (psPT), in this paper, the electroporation of KcsA membrane protein system and bare lipid bilayer system (POPC) with the applied psPT are simulated using molecular dynamics (MD) method. First, we find that the average pore formation time of the KcsA system is longer than the bare system with the applied psPT. In the KcsA system, water protrusions appear more slowly. Then, the system size effects of psPT in the MD simulations are investigated. When the system size decreases, the average pore formation time of small KcsA membrane protein system is shorter than the bare system with the applied psPT. It is found that the psPT makes the protein fluctuation of small system increase greatly; meanwhile the instability of protein disturbs the water and then affects the water protrusion appearance time. Furthermore, it shows that the protein fluctuation of constant electric field is smaller than that of psPT and no field, and protein fluctuation increases with the psPT repetition frequency increasing.

Gut microbiota can affect multiple brain functions and cause behavioral alterations through the microbiota-gut-brain axis. In our previous study, we found that the absence of gut microbiota can influence the expression of microRNAs and mRNAs in the hippocampal region of the germ-free (GF) mice. Long non-coding RNAs (lncRNAs) are increasingly being recognized as an important functional transcriptional regulator in the brain. In the present study, we aim to identify possible biological pathways and functional networks for lncRNA-associated transcript of the gut microbiota in relation to the brain function. The profiles of lncRNA and mRNA from specific pathogen-free (SPF), colonized GF (CGF), and GF mice were generated using the Agilent Mouse LncRNA Array v2.0. Differentially expressed (DE) lncRNAs and mRNAs were identified, and lncRNA target genes were also predicted. Ingenuity pathway analysis (IPA) was performed to analyze related signaling pathways and biological functions associated with these dysregulated mRNAs and target genes. Validation with quantitative real-time PCR was performed on several key genes. Compared with SPF mice a total of 2230 DE lncRNAs were found in GF mice. Among these, 1355 were upregulated and 875 were downregulated. After comparing the target genes of DE lncRNAs with mRNA datasets, 669 overlapping genes were identified. IPA core analyses revealed that most of these genes were highly associated with cardiac hypertrophy, nuclear factors of activated T cells (NFAT) gonadotropin-releasing hormone (GnRH), calcium, and cAMP-response element-binding protein (CREB) signaling pathways. Additionally, mRNA expression levels of APP, CASP9, IGFBP2, PTGDS, and TGFBR2 genes that are involved in central nervous system functions were significantly changed in the GF mouse hippocampus. Through this study, for the first time, we describe the effect of gut microbiota on the hippocampal lncRNA regulation. This will help in enhancing the overall knowledge about microbiota-gut-brain axis.

Key Message: The qHTB1-1 QTL, controlling heat tolerance at the booting stage in rice, was fine mapped to a 47.1 kb region containing eight candidate genes. Two positional candidate genes showed significant changes in expression levels under heat stress. High-temperature stress at the booting stage has the potential to significantly limit rice production. An interspecific advanced backcrossed population between the Oryza sativa L. cultivar R53 and the wild Oryza rufipogon Griff accession HHT4 was used as the source material to develop a set of chromosome segment introgression lines to elucidate the genetic mechanism of the qHTB1-1 QTL in heat tolerance. A single-chromosome-segment introgression line, IL01-15, was used to develop secondary populations for the mapping of qHTB1-1 on chromosome 1 for heat tolerance at the booting stage. Using the BC 5 F 2 , BC 5 F 3 , and BC 5 F 4 populations, we first confirmed qHTB1-1 and validated the phenotypic effect. The qHTB1-1 QTL explained 13.1%, 16.9%, and 17.8% of the phenotypic variance observed in the BC 5 F 2 , BC 5 F 3 , and BC 5 F 4 generations, respectively. Using homozygous recombinants screened from larger BC 6 F 2 and BC 6 F 3 populations, qHTB1-1 was fine mapped within a 47.1 kb region between markers RM11633 and RM11642. Eight putative predicted genes were annotated in the region, and six genes were predicted to encode expressed proteins. The expression patterns of these six genes demonstrated that LOC&#95;Os01g53160 and LOC&#95;Os01g53220 were highly induced by heat stress in IL01-15 compared to R53. Sequence comparison of the gene-coding regions of LOC&#95;Os01g53160 and LOC&#95;Os01g53220 between R53 and IL01-15 revealed one synonymous and two nonsynonymous SNPs in exons, respectively. Our results provide a basis for identifying the genes underlying qHTB1-1 and indicate that markers linked to the qHTB1-1 locus can be used to improve the heat tolerance of rice at the booting stage by marker-assisted selection.

Previously, AoamyA, the alpha amylase-encoding gene from Aspergillus oryzae, was heterologously expressed in Monascus ruber CICC41233 to promote starch hydrolysis and increase the production of Monascus pigments. The target of this study is to screen the effective alpha-amylases from M. ruber for starch fast degradation and investigated for Monascus pigments production. The 13 types of predicted alpha-amylases in the M. ruber NRRL1597 genome were divided into four classes based on EC number and into five groups based on the glycoside hydrolase sub-family. The predicted alpha-amylases MrAMY1 (protein ID 440333) and MrAMY2 (protein ID 324551) showed the closest match with AOamyA by phylogenetic analysis. The genes encoding alpha-amylase, Mramy1 and Mramy2, were cloned from M. ruber CICC41233. However, the gene sequence of Mramy1 from M. ruber CICC41233 differed from that of M.ruber NRRL1597 in the length of the intron sequence. Furthermore, the Mramy1- overexpressed strain M.ruber 440333-6A completely degraded the starch of rice grain in 2 d; in contrast, starch (40.32 mg/mL) remained when rice grain was incubated with the Mramy2- overexpressed strain, M. ruber 324551-D even after 2 d, while 45.43 mg/mL and 10.48 mg/mL starch remained after 2 d and 6 d, respectively, in wild type M. ruber CICC41233. Compared to that of M. ruber CICC41233, the total Monascus pigments and ethanol-soluble pigments in M.ruber 440333-6A increased by 71.69% and 119.33% after 6d, respectively; however, it decreased by 21.40%and 26.58% after 6d, respectively, in M. ruber 324551-D. This study demonstrated that alpha-amylase MrAMY1 was superior to MrAMY2, as it effectively degraded the starch of rice grain and enhanced Monascus pigments production., Competing Interests: Conflict of interestThe authors declare that they have no competing interests., (© King Abdulaziz City for Science and Technology 2020.)

Bactrocera cucurbitae (Coquillett) (Diptera: Tephritidae) is an important pest of vegetables in Asia, the Middle East, Africa, and Hawaii. High temperature can significantly influence B. cucurbitae reproduction. The effect of short-term high-temperature exposure on proteins that affect oviposition was analyzed by proteomics. Among six key target genes for oviposition, the expression of Vitellogenin-1, Vitellogenin-2, and Vitellogenin receptor was similar in B. cucurbitae exposed to higher temperature compared to controls. However, levels of Vitellogenin-3 were reduced. Juvenile hormone (Jh)-inducible protein was downregulated and then upregulated, while the expression of Jh-epoxide hydrolase-2 showed the opposite Jh-inducible protein trend. Therefore, short-term high-temperature stress can cause differential expression of proteins related to oviposition in B. cucurbitae, which in turn further triggers the hormesis of oviposition. High-temperature conditions have become more frequent because of climate warming and are predicted to continue. The data indicate that climate effects on insect reproduction pose a significant threat to agriculture in a world of increasing population.

To investigate the relationship between starch hydrolysis and Monascus pigments (MPs) production, the α-amylase gene ( AOamyA ) from Aspergillus oryzae was heterologously expressed in Monascus ruber CICC41233, and we obtained a positive transformant named Monascus ruber Amy9. In M. ruber Amy9, the α-amylase activities were 6.65- and 4.26-fold higher at 72 h and 144 h, respectively, than those in the parent strain with the glucose as solo carbon medium. Surprisingly, in the MPs fermentation medium with rice powder as solo material, M. ruber Amy9 completely degraded starch at 48 h, while 43.93 and 7.29 mg/mL starch remained at 48 and 144 h, respectively, in the parent strain. Monascus ruber Amy9 accelerated starch hydrolysis, which enhanced biomass and also increased total MPs by 132% after 144 h. Compared with M. ruber CICC41233, the relative gene expression levels, as determined by a quantitative real-time polymerase chain reaction analysis, of acl2 encoding ATP-citrate lyase subunit 2, pks encoding polyketide synthase, and fasB encoding the fatty acid synthase beta subunit increased by 33.14, 145.18, and 32.15%, respectively, after 144 h in M. ruber Amy9. The up-regulated expression of these key genes in MPs synthesis contributed to the large increase in MPs production. This interesting work provided us with a new idea and a new target for the study of the MPs production., Competing Interests: Compliance with ethical standardsThe author declares that they have no conflict of interest.

A hydrothermal method for synthesizing ultralong and thin copper nanowires (CuNWs) with average diameter of 35 nm and average length of 100 μm is demonstrated in this paper. The concerning raw materials include copric (II) chloride dihydrate (CuCl 2 ·2H 2 O), octadecylamine (ODA), and ascorbic acid, which are all very cheap and nontoxic. The effect of different reaction time and different molar ratios to the reaction products were researched. The CuNWs prepared by the hydrothermal method were applied to fabricate CuNW transparent conductive electrode (TCE), which exhibited excellent conductivity-transmittance performance with low sheet resistance of 26.23 [Formula: see text] and high transparency at 550 nm of 89.06% (excluding Polyethylene terephthalate (PET) substrate). The electrode fabrication process was carried out at room temperature, and there was no need for post-treatment. In order to decrease roughness and protect CuNW TCEs against being oxidized, we fabricated CuNW/poly(methyl methacrylate) (PMMA) hybrid TCEs (HTCEs) using PMMA solution. The CuNW/PMMA HTCEs exhibited low surface roughness and chemical stability as compared with CuNW TCEs.

The High osmolarity glycerol (HOG) gene family plays crucial roles in various developmental and physiological processes in fungi, such as the permeability of cell membrane, chlamydospore formation and stress signaling. Although the function of HOG genes has been investigated in Saccharomyces cerevisiae and some filamentous fungi, a comprehensive analysis of HOG gene family has not been performed in Aspergillus oryzae, a fungi mainly used for the production of soy sauce. In this study, we identified and corrected a total of 90 HOG genes from the A. oryzae genome. According to the phylogenetic relationship, they were divided into four discrete groups (Group A-D) comprising of 16, 24, 30 and 20 proteins, respectively. Six conserved motifs and exon-intron structures were examined among all HOG proteins to reveal the diversity of AoHOG genes. Based on transcriptome technology, the expression patterns of AoHOG genes across all developmental stages was identified, suggesting that the AoHOG gene family mainly functions in the logarithmic phase of development. The expression profiles of AoHOG genes under different concentrations of salt stress indicated that AoHOG genes are extensively involved in salt stress response, with possibly different mechanisms. The genome-wide identification, evolutionary, gene structures and expression analyses of AoHOG genes provide a comprehensive overview of this gene family as well as their potential involvements in development and stress responses. Our results will facilitate further research on HOG gene family regarding their physiological and biochemical functions.

The present study verified whether acyl-coenzyme A (acyl-CoA)-binding protein (ACBP) affected the production of Monascus pigments (MPs) in Monascus ruber CICC41233 (MrACBP). Phylogenetic analysis revealed that the cloned Mracbp gene, which encoded the MrACBP protein, exhibited the closest match (99% confidence level) to the gene from Penicilliopsis zonata . The MrACBP and maltose-binding protein (MBP) were simultaneously expressed in Escherichia coli Rosetta DE3 in the form of a fusion protein. The microscale thermophoresis binding assay revealed that the purified MBP-MrACBP exhibited a higher affinity for myristoyl-CoA (Kd = 88.16 nM) than for palmitoyl-CoA (Kd = 136.07 nM) and octanoyl-CoA (Kd = 270.9 nM). Further, the Mracbp gene was homologously overexpressed in M. ruber CICC41233, and a positive transformant M. ruber ACBP5 was isolated. The fatty acid myristic acid in M. ruber ACBP5 was lower than that in the parent strain M. ruber CICC41233. However, when compared with the parent strain, the production of total MPs, water-soluble pigment, and ethanol-soluble pigment in M. ruber ACBP5 increased by 11.67, 9.80, and 12.70%, respectively, after 6 days. The relative gene expression level, as determined by a quantitative real-time polymerase chain reaction analysis, of the key genes acbp , pks , mppr1 , fasA , and fasB increased by 4.03-, 3.58-, 1.67-, 2.11-, and 2.62-fold after 6 days. These data demonstrate the binding preference of MrACBP for myristoyl-CoA, and its influence on MPs production., Competing Interests: Compliance with ethical standardsThe authors declare no conflicts of interest.

Purpose: Current markerless registration methods for neurosurgical robotics use the facial surface to match the robot space with the image space, and acquisition of the facial surface usually requires manual interaction and constrains the patient to a supine position. To overcome these drawbacks, we propose a registration method that is automatic and does not constrain patient position., Methods: An optical camera attached to the robot end effector captures images around the patient's head from multiple views. Then, high coverage of the head surface is reconstructed from the images through multi-view stereo vision. Since the acquired head surface point cloud contains color information, a specific mark that is manually drawn on the patient's head prior to the capture procedure can be extracted to automatically accomplish coarse registration rather than using facial anatomic landmarks. Then, fine registration is achieved by registering the high coverage of the head surface without relying solely on the facial region, thus eliminating patient position constraints., Results: The head surface was acquired by the camera with a good repeatability accuracy. The average target registration error of 8 different patient positions measured with targets inside a head phantom was [Formula: see text], while the mean surface registration error was [Formula: see text]., Conclusion: The method proposed in this paper achieves automatic markerless registration in multiple patient positions and guarantees registration accuracy inside the head. This method provides a new approach for establishing the spatial relationship between the image space and the robot space.

The goal of this study was to enhance the production of xylooligosaccharides (XOs) and reduce the production of xylose. We investigated β-xylosidases, which were key enzymes in the hydrolysis of xylan into xylose, in Trichoderma orientalis EU7-22. The binary vector pUR5750G/ bxl :: hph was constructed to knock out the β - xyl1 gene (encoding β-xylosidases) in T. orientalis EU7-22 by homologous integration, producing the mutant strain T. orientalis Bxyl-1. Xylanase activity for strain Bxyl-1 was 452.42 IU/mL, which increased by only 0.07% compared to that of parental strain EU7-22, whereas β-xylosidase activity was 0.06 IU/mL, representing a 91.89% decrease. When xylanase (200 IU/g xylan), produced by T. orientalis EU7-22 and T. orientalis Bxyl-1, was used to hydrolyze beechwood xylan, in contrast to the parental strain, the XOs were enhanced by 83.27%, whereas xylose decreased by 45.80% after 36 h in T. orientalis Bxyl-1. Based on these results, T. orientalis Bxyl-1 has great potential for application in the production of XOs from lignocellulosic biomass., Competing Interests: Compliance with ethical standardsThe authors declare no conflicts of interest.

The role of the transcription factor creA-mediating carbon catabolite repression in Trichoderma orientalis EU7-22 was investigated for cellulase and hemicellulase production. The binary vector pUR5750G/creA::hph was constructed to knock out creA by homologous integration, generating the ΔcreA mutant Trichoderma orientalis CF1D. For strain CF1D, the filter paper activities (FPA), endoglucanase activities (CMC), cellobiohydrolase activity(CBH), β-glucosidase activity (BG), xylanase activity (XYN), and extracellular protein concentration were 1.45-, 1.15-, 1.71-, 2.51-, 2.72, and 1.95-fold higher in inducing medium and were 6.41-, 7.50-, 10.27-, 11.79-, 9.25-, and 3.77-fold higher in glucose repressing medium, respectively, than those in the parent strain after 4 days. SDS-PAGE demonstrated that the extracellular proteins were largely secreted in the mutant CF1D. Quantitative reverse-transcription polymerase chain reaction indicated that the expressions of cbh1, cbh2, eg1, eg2, bgl1, xyn1, and xyn2 were significantly increasing for the mutant CF1D not only in the inducing medium but also in the repressing medium. Those results indicated that creA was a valid target gene in strain engineering for improved enzyme production in T. orientalis.

Purpose: Using existing stereoelectroencephalography (SEEG) electrode implantation surgical robot systems, it is difficult to intuitively validate registration accuracy and display the electrode entry points (EPs) and the anatomical structure around the electrode trajectories in the patient space to the surgeon. This paper proposes a prototype system that can realize video see-through augmented reality (VAR) and spatial augmented reality (SAR) for SEEG implantation. The system helps the surgeon quickly and intuitively confirm the registration accuracy, locate EPs and visualize the internal anatomical structure in the image space and patient space., Methods: We designed and developed a projector-camera system (PCS) attached to the distal flange of a robot arm. First, system calibration is performed. Second, the PCS is used to obtain the point clouds of the surface of the patient's head, which are utilized for patient-to-image registration. Finally, VAR is produced by merging the real-time video of the patient and the preoperative three-dimensional (3D) operational planning model. In addition, SAR is implemented by projecting the planning electrode trajectories and local anatomical structure onto the patient's scalp., Results: The error of registration, the electrode EPs and the target points are evaluated on a phantom. The fiducial registration error is [Formula: see text] mm (max 1.22 mm), and the target registration error is [Formula: see text] mm (max 1.18 mm). The projection overlay error is [Formula: see text] mm, and the TP error after the pre-warped projection is [Formula: see text] mm. The TP error caused by a surgeon's viewpoint deviation is also evaluated., Conclusion: The presented system can help surgeons quickly verify registration accuracy during SEEG procedures and can provide accurate EP locations and internal structural information to the surgeon. With more intuitive surgical information, the surgeon may have more confidence and be able to perform surgeries with better outcomes.