bluuebunny/crossref_metadata_2025_split huggingface dataset:
High-Throughput Screening of Higher Α-Amylase Producing Bacillus Licheniformis by Fluorescence-Activated Droplet Sorting
Authors: Yuan Huiling , Tu Ran , Tong Xinwei , Lin Yuping , Wang Qinhong 
Published: July 2021

Abstract
Backgroundα-Amylases is one of the most important starch degrading enzymes and has the widest range of industrial applications. Bacillus licheniformis has been widely used as a cell factory for industrial production of amylase. However, difficulties in genetic modification of B. licheniformis have limited its widespread use. Directed evolution, based on the combination of random mutagenesis and high throughput screening (HTS), has been proven an effective strategy in strain improvement for increasing the productivity, but it requires a suitable HTS system to screen the desired mutants. Droplet-based microfluidics has emerged as a powerful tool for single-cell screening with ultra-high throughput, however, the accessibility of a droplet microfluidic HTS platform to users having no background in microfluidics is still an issue. ResultsHere, we first developed a microfluidic HTS platform based on fluorescence-activated droplet sorting (FADS) technology. This platform allowed (i) encapsulation of single cells in monodisperse water-in-oil droplets; (ii) cell growth and protein production in droplets; (iii) sorting of droplets based on their fluorescence intensities. To validate the platform, a model selection experiment of a binary mixture of Bacillus strains was performed and a 45.6-fold enrichment was achieved at a sorting rate of 300 droplets per second. Furthermore, we used the platform for the selection of higher α-amylase-producing strains from a library of B. licheniformis strains (a strain already used at industrial-scale). The B. licheniformis mutant library was generated by atmospheric and room temperature plasma (ARTP) mutagenesis. The clones displaying over 50% improvement in α-amylases productivity compared to the wild-type were isolated.ConclusionsWe established an efficient droplet microfluidic platform which consisted of droplet generation, droplet incubation, and sorting of droplets with a throughput of up to 1 × 106 droplets per h. The screening platform was demonstrated by successfully identifying B. licheniformis clones with improved α-amylase production. We believe that the droplet platform could be extended to the development of other industrially valuable strains.
Bifurcation Functional Significance Score as Predictor of Mortality
Authors: Vassilev Dobrin , Mileva Niya , Collet Carlos , Nikolov Pavel , Sokolova Katerina , Karamfiloff Kiril , Naunov Vladimir , Sonck Jeroen , Rigatelli Gianluca , Kassab Ghassan S. , Gil Robert J. 
Published: July 2021

Abstract
Background: Considerable progress has been made in the treatment of coronary bifurcation stenosis. Anatomical characteristics of the vessel and lesion, however, fail to give information about the functional significance of the bifurcation stenosis. To the best of our knowledge, there is no study that systematically establishes the baseline functional significance of coronary stenosis and its effect on procedural and clinical outcomes.Methods: Patients with significant angiographic bifurcation lesions defined as diameter stenosis > 50% in main vessel and/or side branch were included. FFR was performed in main vessel (MV) and side branch (SB) before and after percutaneous coronary intervention (PCI). 169 patients from Fiesta study (derivation cohort) and 555 patients from prospective bifurcation registry (test cohort) were analyzed to validate angiographic prediction score (BFSS) used to determine the potentially functional significance of coronary bifurcation stenosis.Results: Bifurcation functional significance score (including the following parameters - SYNTAX ≥11, SB/MB BARI score, MV %DS ≥ 55%, MB %DS ≥ 65%, lesion length ≥25mm) with a maximum value of 11 was developed. A cut-off value of 8.5 was shown to give the best discriminatory ability – with accuracy 87% (sensitivity 77%, specificity 96%, p<.001). There was also a significant difference in all-cause mortality between patients with BFSS≥8.5 vs. BFSS<8.5 – 17.8% vs. 13.4%, log-rank p=.006, as well as cardiac mortality: BFSS≥8.5 vs. BFSS<8.5 – 14.6% vs. 12.5%, log-rank (p=.042). The cardiac mortality was significantly lower in patients with smaller absolute SB territory, p=0.023.Conclusion: An angiographic score (BFSS) with good discriminatory ability to determine the functional significance of coronary bifurcation stenosis was developed. The value for BFSS≥8.5 can be used as a discriminator to define groups with higher risk for all-cause and cardiac mortality. Also, we found that the smaller side branches pose greater mortality risk.
A Safety Response Mechanism for an Autonomous Moving Robot in a Small Manufacturing Environment using Q-learning Algorithm and Speech Recognition
Authors: KIANGALA Sonia , Wang Zenghui 
Published: July 2021

Abstract
The industrial manufacturing sector is currently undergoing a tremendous revolution moving from traditional production processes to intelligent techniques. Under this revolution, known as Industry 4.0 (I40), a robot is no longer a static equipment but an active workforce to the factory production alongside human operators. Safety becomes crucial for humans and robots to ensure a smooth production run in such environments. Operators are subject to frequent safety inductions to react in emergencies but very little is done for robots. Our research proposes a safety response mechanism for a small manufacturing plant, through which an autonomous robot learns the obstacle-free trajectory to the closest safety exit in emergencies. We implement a reinforcement learning (RL) algorithm, Q-learning, to enable the path learning abilities of the robot. After obtaining the robot optimal path selection options with Q-learning, we code the outcome as a rule-based system for the safety response. We also program a speech recognition system for operators to react timeously, with a voice command, to an emergency that requires stopping all plant activities even when they are far away from emergency stops (ESTOPs) button. The factory emergency signal can be given by an ESTOP or a voice command sent directly to the factory central controller: an S7-1200 Siemens programmable logic controller (PLC) in this experiment. We simulate a simple and small manufacturing environment overview to test our safety procedure. Our results show that the safety response mechanism successfully generates paths without obstacles to the closest safety exits from all the factory locations.
Bullatine a Exerted Anti-Inflammatory Effects by Inhibiting JNK/ROS/NF-κB Pathway and Attenuates Systemic Inflammatory Response in LPS-Challenged Mice
Authors: Liu Shuhan , Yan Meichen , Liao Yajin , Cheng Yong 
Published: July 2021

Abstract
Background: The genus Aconitum has rich pharmacological characteristics. Aconiti brachypodi Radix (Xue-shang-yi-zhi-hao) is a dried root of aconitum, which is considered to be analgesic and anti-inflammatory in modern medical and pharmaceutical clinical studies. Bullatine A (BA), a major active ingredient of this plant, has been reported for its significant anti-analgesic effect in previous studies. However, the role of BA in inflammation is unknown. In the current study, we aimed to explore the effect of BA on lipopolysaccharide (LPS)-induced inflammatory response both in vitro and in vivo and its potential anti-inflammatory mechanism.Materials and Methods: The anti-inflammatory effect of BA was evaluated in two different types of LPS-induced macrophages, including BV-2 microglial cells and immortalized murine bone marrow-derived macrophages (iBMDMs), and in acute inflammation mouse models induced by LPS. Immunofluorescence, flow cytometry, quantitative RT-PCR, western blot and Hematoxylin-Eosin staining were used to determine the anti-inflammatory properties of BA.Results: The results showed that BA significantly reduced the mRNA levels of several pro-inflammatory cytokines induced by LPS both in BV-2 cells and iBMDMs. Inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in response to LPS were also decreased by BA. Further investigations indicated BA significantly blocked the phosphorylation of IκB kinase, degradation of the inhibitor IκBa and the nuclear translocation of nuclear factor-κB (NF-κB) p65. BA also reduced c-Jun N-terminal kinases (JNK) phosphorylation and ROS generation in iBMDMs activated with LPS, but had no effect on other mitogen-activated protein kinases (MAPKs) family proteins such as extracellular signal-regulated kinase (ERK) or p38. Furthermore, BA treatment alleviate liver and lung tissue damage, reduce inflammatory cell infiltration, and inhibit the expression of inflammatory cytokines in LPS-challenged mice.Conclusions: This study illustrated that BA has obvious anti-inflammatory effects both in vitro and in vivo, and its underlying anti-inflammatory mechanism may be via inactivating JNK/ROS/NF-κB pathway. Therefore, BA may have a certain therapeutic potential for inflammatory-related diseases.
Determination of Exosome Mitochondrial DNA as a Biomarker of Renal Cancer Aggressiveness
Authors: Arance Elena , Ramírez Viviana , Rubio-Roldan Alejandro , Ocaña-Peinado Francisco M. , Romero-Cachinero Catalina , Vazquez-Alonso Fernando , Martinez-Gonzalez Luis Javier , Alvarez-Cubero Maria Jesus 
Published: July 2021

Abstract
Background: Components of liquid biopsy are promising non-invasive biomarkers for monitoring the renal cell carcinoma (RCC) status. Present study tries to evaluate the role of exosomes and mitochondrial DNA (mtDNA) as promise, novel and stable biomarkers that could improve current ones in RCC. Methods: A total of 140 fractions obtained by ultracentrifugations of whole blood samples stored in EDTA anticoagulant from 28 (13 patients and 15 controls) were included in present study. An exosome collection protocol and exhaustive analysis of all phases obtained by ultracentrifugations (named from B to F) was performed. Subsequently, an analysis of dPCR (digital PCR) using the QuantStudio™ 3D Digital PCR platform and the quantification of mtDNA copy number using by QuantStudioTM 12K Flex Real-Time PCR System (qPCR) was developed. Moreover, Next Generation Sequencing (NGS) analyses were included using MiSeq system (Illumina, CA, USA). Results: F fraction, which contains all exosomes´ data and all mitochondrial markers (hypervariable region 1 (HV1) and apocytochrome b of complex III (MT-CYB)) were statistically identified in dPCR and qPCR (p values < 0.05) when a comparison between cases and controls was performed. Moreover, the studied mitochondrial genes shown a relevant significance in RCC aggressiveness analyses (metastasis and stages 4).Conclusions: To sum up, this is the first time a relation between mtDNA genetic markers in exosome and clinical management of RCC is suggested.
Decadal Variation of the Precipitation Relationship Between June and August over South China and Its Mechanism
Authors: li shuai , Gong Zhiqiang , zhang Shixuan , Yang Jie , Qiao Shaobo , Feng Guolin 
Published: July 2021

Abstract
This paper investigates the characteristics and causes for the interdecadal change in the relationships between early and late summer rainfall over South China (SC). This study finds that the correlations of the precipitation over SC between June and August shift from weakly positive in 1979 – 1995 to obviously negative in 1996-2019. Further analysis demonstrates that the interdecadal variations of monthly SST anomaly (SSTA) and associated air-sea interactions in June and August account for the decadal variations of the precipitation relationships. During the prior period 1979-1995, the tropical West Indian Ocean (WIO) shows a significant positive SSTA in June, which triggers Kevin waves and an anticyclone circulation over the tropical Northwest Pacific (NWP). The warm and wet air transported by the southwest airflow at the north of the anticyclone provides favorable environmental condition to produce more precipitation over SC region in June. In contrast, the SST dipole pattern with the negative SSTAs in the maritime continent (MC) and positive SSTAs in the tropical Central Pacific (CP) is dominant in August. The SST dipole pattern is inconducive to the formation of anticyclone over SC, causing a weak positive precipitation correlation between June and August. During the latter period 1996-2019, the precipitation over SC in June is the same as that in the prior period as there is no significant decadal change in tropical WIO SST and East Asian circulation. However, an opposite phase of the SST dipole anomaly pattern in MC and the tropical CP is dominant in August during the latter period. Accordingly, the positive feedback mechanism of air-sea interaction leads to the enhancement of local convection activities in MC and the meridional Hadley circulations and the NWP subtropical high, leading to a decrease of precipitation over SC in August. Overall, the decadal variation of the SST dipole anomaly pattern in MC and the tropical CP is the key factor affecting the adjustment of the correlations between June and August precipitation in the two periods.
The Rigorous Screening and Ethics of Donor for Faecal Microbiota Transplantation in China
Authors: He Jianquan , He Xingxiang , Ma Yonghui , Yang Luxi , Fang Haiming , Shang Shu , Xia Huping , Lian Guanghui , Tang Hailing , Wang Qizhi , Wang Junping , Lin Zhihui , Wen Jianbo , Liu Yuedong , Zhai Chunbao , Wang Wen , Jiang Xueliang , Xuan Ji , Liu Morong , Lu Shiyun , Li Xuejun , Wang Han , Ouyang Cong , Cao Man , Lin Aiqiang , Zhang Bangzhou , Wu Depei , Chen Ye , xiao chuanxing 
Published: July 2021

Abstract
Background: Faecal microbiota transplantation (FMT) is a safe and effective therapy for recurrent Clostridium difficile infections and some gastrointestinal and systemic diseases. However, the risks of FMT and the selection process of suitable donors remain insufficiently characterized. The eligibility rate for screening, underlying microbial basis, and core ethical issues of stool donors for FMT are yet to be elucidated in China. Results：The potential stool donors were screened from December 2017 to December 2019 with the help of an online survey, clinical assessments, and stool and blood testing. Bioinformatics analyses were performed, and the composition and stability of gut microbiota in stool obtained from eligible donors were dynamically observed using metagenomics. Meanwhile, we build a donor microbial evaluation index (DoMEI) for stool donor screening. In the screening process, we also focused on ethical principles and requirements. Of the 2071 participants, 66 donors were selected via the screening process (3.19% success rate). Although there were significant differences in gut microbiota among donors, we found that the changes in the gut microbiota of the same donor were typically more stable than those between donors over time. Conclusions: The DoMEI provides a potential reference index for stool donor re-evaluation. In this retrospective study, we summarised the donor recruitment and screening procedure for FMT in China. Based on the latest advances in this field, we carried out a rigorous faecal microbiota donor screening process.
Combined Hormone Receptor and Human Epidermal Growth Factor Receptor-2 Expression Reflects Inter-and Intra-tumor Heterogeneity in Hormone Receptor-Positive/Human Epidermal Growth Factor Receptor-2-Positive Breast Cancer
Authors: Ju Jie , Du Feng , Gao Song-Lin , Si Yi-Ran , Hu Nan-Lin , Liu Dong-Xu , Wang Xue , Yue Jian , Zheng Fang-Chao , Kang Yi-Kun , Yang Zi-Xuan , Ma Fei , Xu Bing-He , Yuan Peng 
Published: July 2021

Abstract
BackgroundHormone receptor-positive and human epidermal growth factor receptor-2-positive (HR+/HER2+) breast cancer comprise approximately 5-10% of all invasive breast cancers. However, the lack of knowledge regarding the complexity of tumor heterogeneity in HR+/HER2+ disease remains a barrier to more accurate therapies. This study aimed to describe the tumor heterogeneity of HR+/HER2+ breast cancer and to establish a novel indicator to identify the HER2-enriched-like subtype in patients with HR+/HER2+ breast cancer.MethodsFirst, a comprehensive analysis was performed on HR+/HER2+ breast cancer samples from the TCGA (n=141) and METABRIC (n=104) databases. We determined the distribution of PAM50 intrinsic subtypes within the two cohorts and compared the somatic mutational profile and RNA expression features between HER2-enriched and non-HER2-enriched subtypes. We then performed multiplex immunofluorescence to evaluate HER2 and estrogen receptor (ER) expression simultaneously in the third cohort, enrolling 43 cases of early HR+/HER2+ breast cancer from Cancer Hospital, Chinese Academy of Medical Sciences (CAMS).ResultsAll four main intrinsic subtypes were identified in HR+/HER2+ breast cancer, of which the luminal-B subtype was the most common, followed by the HER2-enriched and luminal-A subtypes. Significantly increased TP53 and ERBB3 and decreased PIK3CA somatic mutation frequency were observed in the HER2-enriched subtype compared with the non-HER2-enriched subtype. In addition,the HER2-enriched subtype was characterized by significantly higher ERBB2 and lower ESR1 expression. We then constructed a marker termed rH/E to reflect the relative expression of ERBB2 to ESR1 in each patient.rH/E discriminates the HER2-enriched subtype from the non-HER2 subtypes better than the expression of ERBB2 or ESR1 alone. In the CAMS cohort, we observed four subtypes of tumor cells: ER+/HER2-,ER+/HER2+,ER-/HER2+, and ER-/HER2-. Tumor cell diversity was common, with 86% of patients having all four subtypes of tumor cells. Moreover, rH/E showed a significant prognostic association in the CAMS cohort.ConclusionsThis study furthers our understanding of the complexity of tumor heterogeneity in HR+/HER2+breast cancer, and suggests that the combined analysis of ERBB2 and ESR1 expression may contribute to identifying patients with specific subtypes in this population.
Assessment of the Relationship Between Labial Gingival Thickness and Underlying Bone Thickness in Maxillary Anterior Teeth by Two Digital Techniques
Authors: Wang Linhong , Chen Jianping , Ruan Yan , Luo Yunxiao , Yang Fan 
Published: July 2021

Abstract
The aim of this study is to noninvasively assess the relationship between the labial gingival thickness (GT) and the underlying bone thickness (BT) of the maxillary anterior teeth by two digital techniques. GT and BT were measured at 2、4 and 6 mm apical to the cemento-enamel junction (CEJ) by two digital techniques: M1- cone-beam computed tomography (CBCT) and M2- digital intraoral scanning (DIS) combined with CBCT (30 healthy participants, 172 maxillary anterior teeth). Pearson's correlation coefficient was calculated to assess the correlation between GT and BT. A significant negative correlation was identified between GT and BT at 2、4 mm apical to the CEJ for central incisors (CI), lateral incisors (LI), and canines (CA) both by M1 and M2, but not at 4 mm apical to the CEJ for CA (M2) or at 6 mm apical to the CEJ for all (M1, M2). The labial BT was demonstrated < 1 mm in most cases (85% of the CI; 97% of the LI; and 90% of the CA). Within the limitation of this study, it is concluded that the GT and BT seems to be negatively correlated in most cases, which should be cautious clinically.
Prognostic Modeling of Patients with Metastatic Melanoma Based on Tumor Immune Microenvironment Characteristics
Authors: Liu Jing , Ye Ting , Zhang Xue fang , Dong Yong jian , Zhang Wen feng , Wu Feng lin , Bo Hua ben , Shao Hong wei , Zhang Rong xin , Shen Han 
Published: July 2021

Abstract
Most of the malignant melanomas are already in the middle and advanced stages when they are diagnosed, which is often accompanied by the metastasis and spread of other organs.Besides, the prognosis of patients is bleak. The characteristics of the local immune microenvironment in metastatic melanoma have important implications for both tumor progression and tumor treatment. In this study, data on patients with metastatic melanoma from the TCGA and GEO datasets were selected for immune, stromal, and estimate scores, and overlapping differentially expressed genes (DEGs) were screened. A nine-IRGs prognostic model (ALOX5AP, ARHGAP15, CCL8, FCER1G, GBP4, HCK, MMP9, RARRES2 and TRIM22) was established by univariate COX regression, LASSO and multivariate COX regression. Receiver operating characteristic (ROC) curves were used to test the predictive accuracy of the model. Immune infiltration was analyzed by using CIBERSORT, Xcell and ssGSEA in high-risk and low-risk groups. The immune infiltration of the high-risk group was significantly lower than that of the low-risk group. Immune checkpoint analysis revealed that the expression of PDCD1, CTLA4, TIGIT, CD274, HAVR2 and LAG3 were significantly different in groups with different levels of risk scores. WGCNA analysis found that the yellow-green module contained seven genes (ALOX5AP, FCER1G, GBP4, HCK, MMP9, RARRES2 and TRIM22) from the nine-IRG prognostic model, of which the yellow-green module had the highest correlation with risk scores. The results of GO and KEGG suggested that the genes in the yellow-green module were mainly enriched in immune-related biological processes. Finally, we analyzed the prognostic ability and expression characteristics of ALOX5AP, ARHGAP15, CCL8, FCER1G, GBP4, HCK, MMP9, RARRES2 and TRIM22 in metastatic melanoma. Overall, a prognostic model for metastatic melanoma based on the characteristics of the tumor immune microenvironment was established, which was helpful for further studies.It could function well in helping people to understand the characteristics of the immune microenvironment in metastatic melanoma and to find possible therapeutic targets.
Visualisation and measurement of a laminar boundary layer on the surface of an isothermal section-triangular roof
Authors: Zhai Haoyu , Torres Juan F. , Zhao Yongling , Xu Feng 
Published: July 2021

Abstract
Natural convection in air over a heated section-triangular roof with a fixed aspect ratio of 0.1 is experimentally investigated. The development of the flow over the roof subject to a range of temperatures is measured by digital interferometry and thermocouples. The experiments present distinct images of the thermal boundary layer, which changes from a quasi-steady to an unsteady state as the surface temperature of the triangular roof increases. Contrary to numerical simulations previously published, the observed flow becomes unsteady, which is very likely influenced by uncontrolled perturbations at the critical Rayleigh number where a pitchfork bifurcation of a steady flow is theoretically expected.