bluuebunny/crossref_metadata_2025 huggingface dataset:
A Pilot Study of the Efficacy and Economical Sustainability of Acute Coronavirus Disease 2019 Patient Management in an Outpatient Setting
Authors: De Lorenzo Rebecca , Montagna Marco , Bossi Eleonora , Vitali Giordano , Taino Alba , Cilla Marta , Pata Giulia , Lazorova Ludmilla , Pesenti Riccardo , Pomaranzi Chiara , Bussolari Cecilia , Martinenghi Sabina , Bordonaro Nicoletta , Di Napoli Davide , Rizzardini Giuliano , Cogliati Chiara , Morici Nuccia , Rovere-Querini Patrizia
Published: April 2022

Objective

To report a preliminary experience of outpatient management of patients with Coronavirus disease 2019 (COVID-19) through an innovative approach of healthcare delivery.

Patients and Methods

Patients evaluated at the Mild-to-Moderate COVID-19 Outpatient clinics (MMCOs) of San Raffaele University Hospital and Luigi Sacco University Hospital in Milan, Italy, from 1 October 2020 to 31 October 2021 were included. Patients were referred by general practitioners (GPs), Emergency Department (ED) physicians or hospital specialists (HS) in case of moderate COVID-19. A classification and regression tree (CART) model predicting ED referral by MMCO physicians was developed to aid GPs identify those deserving immediate ED admission. Cost-effectiveness analysis was also performed.

Results

A total of 660 patients were included. The majority (70%) was referred by GPs, 21% by the ED and 9% by HS. Patients referred by GPs had more severe disease as assessed by peripheral oxygen saturation (SpO2), ratio of arterial oxygen partial pressure to fractional inspired oxygen (PaO2/FiO2), C-reactive protein (CRP) levels and interstitial involvement at lung ultrasound. Among them, 18% were addressed to the ED following MMCO assessment. CART analysis identified three independent predictors, namely home-measured SpO2, age and body mass index (BMI), that robustly divide patients into risk groups of COVID-19 severity. Home-measured SpO2 < 95% and BMI ≥ 33 Kg/m2 defined the high-risk group. The model yielded an accuracy (95% CI) of 83 (77–88)%. Outpatient management of COVID-19 patients allowed the national healthcare system to spare 1,490,422.05 € when compared with inpatient care.

Conclusion

Mild-to-moderate COVID-19 outpatient clinics were effective and sustainable in managing COVID-19 patients and allowed to alleviate pressure on EDs and hospital wards, favoring effort redirection toward non-COVID-19 patients.
Association Between Trajectory Patterns of Body Mass Index Change Up to 10 Months and Early Gut Microbiota in Preterm Infants
Authors: Qiu Jun , Zhou Changci , Xiang Shiting , Dong Jie , Zhu Qifeng , Yin Jieyun , Lu Xiulan , Xiao Zhenghui
Published: April 2022

Recent research suggests that gut microbiota plays an important role in the occurrence and development of excessive weight and obesity, and the early-life gut microbiota may be correlated with weight gain and later growth. However, the association between neonatal gut microbiota, particularly in preterm infants, and excessive weight and obesity remains unclear. To evaluate the relationship between gut microbiota and body mass index (BMI) growth trajectories in preterm infants, we examined microbial composition by performing 16S rDNA gene sequencing on the fecal samples from 75 preterm infants within 3 months after birth who were hospitalized in the neonatal intensive care unit of Hunan Children’s Hospital from August 1, 2018 to October 31, 2019. Then, we collected their physical growth information during 0–10 months. Latent growth mixture models were used to estimate growth trajectories of infantile BMI, and the relationship between the gut microbiota and the BMI growth trajectories was analyzed. The results demonstrated that there were 63,305 and 61 operational taxonomic units in the higher BMI group (n = 18), the lower BMI group (n = 51), and the BMI catch-up group (n = 6), respectively. There were significant differences in the abundance of the gut microbiota, but no significant differences in the diversity of it between the lower and the higher BMI group. The BMI growth trajectories could not be clearly distinguished because principal component analysis showed that gut microbiota composition among these three groups was similar. The three groups were dominated by Firmicutes and Proteobacteria in gut microbiota composition, and the abundance of Lactobacillus in the higher BMI group was significantly different from the lower BMI group. Further intervention experiments and dynamic monitoring are needed to determine the causal relationship between gut microbiota differences and the BMI change.
Atypical Antibody Dynamics During Human Coronavirus HKU1 Infections
Authors: Sechan Ferdyansyah , Grobben Marloes , Edridge Arthur W. D. , Jebbink Maarten F. , Loens Katherine , Ieven Margareta , Goossens Herman , van Hemert-Glaubitz Susan , van Gils Marit J. , van der Hoek Lia
Published: April 2022

Human coronavirus HKU1 (HCoV-HKU1) is one of the four endemic coronaviruses. It has been suggested that there is a difference in incidence, with PCR-confirmed HCoV-NL63 and HCoV-OC43 infections occurring more commonly, whereas HCoV-HKU1 is the least seen. Lower incidence of HCoV-HKU1 infection has also been observed in serological studies. The current study aimed to investigate antibody dynamics during PCR-confirmed HCoV-HKU1 infections using serum collected during infection and 1 month later. We expressed a new HCoV-HKU1 antigen consisting of both the linker and carboxy-terminal domain of the viral nucleocapsid protein and implemented it in ELISA. We also applied a spike-based Luminex assay on serum samples from PCR-confirmed infections by the four endemic HCoVs. At least half of HCoV-HKU1-infected subjects consistently showed no antibody rise via either assay, and some subjects even exhibited substantial antibody decline. Investigation of self-reported symptoms revealed that HCoV-HKU1-infected subjects rated their illness milder than subjects infected by other HCoVs. In conclusion, HCoV-HKU1 infections reported in this study displayed atypical antibody dynamics and milder symptoms when compared to the other endemic HCoVs.
Achieving Partial Nitritation by Treating Sludge With Free Nitrous Acid: The Potential Role of Quorum Sensing
Authors: Jiang Cancan , Wang Xu , Wang Huacai , Xu Shengjun , Zhang Wei , Meng Qingjie , Zhuang Xuliang
Published: April 2022

Partial nitritation is increasingly regarded as a promising biological nitrogen removal process owing to lower energy consumption and better nitrogen removal performance compared to the traditional nitrification process, especially for the treatment of low carbon wastewater. Regulating microbial community structure and function in sewage treatment systems, which are mainly determined by quorum sensing (QS), by free nitrous acid (FNA) to establish a partial nitritation process is an efficient and stable method. Plenty of research papers reported that QS systems ubiquitously existed in ammonia oxidizing bacteria (AOB) and nitrite oxidizing bacteria (NOB), and various novel nitrogen removal processes based on partial nitritation were successfully established using FNA. Although the probability that partial nitritation process might be achieved by the regulation of FNA on microbial community structure and function through the QS system was widely recognized and discussed, the potential role of QS in partial nitritation achievement by FNA and the regulation mechanism of FNA on QS system have not been reviewed. This article systematically reviewed the potential role of QS in the establishment of partial nitritation using FNA to regulate activated sludge flora based on the summary and analysis of the published literature for the first time, and future research directions were also proposed.
Development of an Indirect ELISA Kit for Rapid Detection of Varicella-Zoster Virus Antibody by Glycoprotein E
Authors: Niu Yan , Wang Aiping , Zhou Jingming , Liu Hongliang , Chen Yumei , Ding Peiyang , Qi Yanhua , Liang Chao , Zhu Xifang , Zhang Gaiping
Published: April 2022

Varicella-zoster virus (VZV), a highly infectious agent that causes varicella (chickenpox), can also cause zoster (shingles), a disorder that is frequently associated with severe neuralgia. A reliable serological VZV diagnostic assay would be useful for identifying unprotected individuals and for surveilling post-vaccination immunoprotection status. Toward this goal, VZV membrane glycoprotein E (gE), the immunodominant VZV protein, served as target antigen in an indirect ELISA kit developed here to detect anti-VZV antibodies in clinical samples. For target antigen preparation, Chinese hamster ovary (CHO) cells were modified to express and secrete the VZV gE ectodomain, which was subsequently purified and used as coating antigen in an indirect ELISA. Ultimately, the optimal purified gE coating antigen concentration was determined to be 2 μg.ml−1 and the OD450nm detection cutoff value was 0.286. The coefficient of variation (CV) of intra-assay and inter-assay were <10 and 15%, respectively. A comparative test of 66 clinical samples showed that the coincidence rate was 93.9% between the indirect ELISA and a commercial varicella-zoster virus IgG ELISA kit. Thus, the indirect ELISA kit developed here may be useful for achieving rapid, sensitive, and specific detection of anti-VZV antibodies.
Genome-wide Characterization of the JmjC Domain-Containing Histone Demethylase Gene Family Reveals GhJMJ24 and GhJMJ49 Involving in Somatic Embryogenesis Process in Cotton
Authors: Li Yan , Zhu Shouhong , Yao Jinbo , Fang Shengtao , Li Tengyu , Li Bei , Wang Xinyu , Wang Mingyang , Wu Lanxin , Pan Jingwen , Feng Xuemei , Chen Wei , Zhang Yongshan
Published: April 2022

The Jumonji C (JmjC) domain-containing protein family, an important family of histone demethylase in plants, can directly reverse histone methylation and play important roles in various growth and development processes. In the present study, 51 JmjC genes (GhJMJs) were identified by genome-wide analysis in upland cotton (Gossypium hirsutum), which can be categorized into six distinct groups by phylogenetic analysis. Extensive syntenic relationship events were found between G. hirsutum and Theobroma cacao. We have further explored the putative molecular regulatory mechanisms of the JmjC gene family in cotton. GhJMJ24 and GhJMJ49 were both preferentially expressed in embryogenic callus compared to nonembryogenic callus in cotton tissue culture, which might be regulated by transcription factors and microRNAs to some extent. Further experiments indicated that GhJMJ24 and GhJMJ49 might interact with SUVH4, SUVH6, DDM1, CMT3, and CMT1 in the nucleus, potentially in association with demethylation of H3K9me2. Taken together, our results provide a foundation for future research on the biological functions of GhJMJ genes in cotton, especially in somatic embryogenesis in cotton tissue culture, which is crucial for the regeneration of transgenic plants.
Bioinformatics Analysis of Publicly Available Single-Nuclei Transcriptomics Alzheimer’s Disease Datasets Reveals APOE Genotype-Specific Changes Across Cell Types in Two Brain Regions
Authors: Belonwu Stella A. , Li Yaqiao , Bunis Daniel G. , Rao Arjun Arkal , Solsberg Caroline Warly , Oskotsky Tomiko , Taubes Alice L. , Grone Brian , Zalocusky Kelly A. , Fragiadakis Gabriela K. , Huang Yadong , Sirota Marina
Published: April 2022

Alzheimer’s Disease (AD) is a complex neurodegenerative disease that gravely affects patients and imposes an immense burden on caregivers. Apolipoprotein E4 (APOE4) has been identified as the most common genetic risk factor for AD, yet the molecular mechanisms connecting APOE4 to AD are not well understood. Past transcriptomic analyses in AD have revealed APOE genotype-specific transcriptomic differences; however, these differences have not been explored at a single-cell level. To elucidate more complex APOE genotype-specific disease-relevant changes masked by the bulk analysis, we leverage the first two single-nucleus RNA sequencing AD datasets from human brain samples, including nearly 55,000 cells from the prefrontal and entorhinal cortices. In each brain region, we performed a case versus control APOE genotype-stratified differential gene expression analysis and pathway network enrichment in astrocytes, microglia, neurons, oligodendrocytes, and oligodendrocyte progenitor cells. We observed more global transcriptomic changes in APOE4 positive AD cells and identified differences across APOE genotypes primarily in glial cell types. Our findings highlight the differential transcriptomic perturbations of APOE isoforms at a single-cell level in AD pathogenesis and have implications for precision medicine development in the diagnosis and treatment of AD.
Facing Your Fear in Immersive Virtual Reality: Avoidance Behavior in Specific Phobia
Authors: Binder Florian P. , Pöhlchen Dorothee , Zwanzger Peter , Spoormaker Victor I.
Published: April 2022

Specific phobias are the most common anxiety disorder and are characterized by avoidance behavior. Avoidance behavior impacts daily function and is proposed to impair extinction learning. However, despite its prevalence, its objective assessment remains a challenge. To this end, we developed a fully automated experimental procedure using immersive virtual reality. The procedure contained a behavioral search, forced-choice, and an approach task with varying degrees of freedom and task relevance of the stimuli. In this study, we examined the sensitivity and feasibility of these tasks to assess avoidance behavior in patients with specific phobia. We adapted the tasks by replacing the originally conditioned stimuli with a spider and a neutral animal and investigated 31 female participants composed of 15 spider-phobic and 16 non-phobic participants. As the non-phobics were quite heterogeneous in terms of their Fear of Spiders Questionnaire (FSQ) scores, we subdivided them into six “fearfuls” that had elevated FSQ scores, and 10 “non-fearfuls” that had no fear of spiders. The phobics successfully managed to complete the procedure and showed consistent avoidance behavior across all behavioral tasks. Compared to the non-fearfuls, which did not show any avoidance behavior at all, the phobics looked at the spider much more often and clearly directed their body toward it in the search task. In the approach task, they hesitated most when they were close to the spider, and their difficulty to touch the spider was reflected in a strong increase in right hand acceleration changes. The fearfuls showed avoidance behavior depending on the tasks: strongest in the search task and weakest in the approach task. Additionally, we identified subjective valence ratings of the spider as the main influence on both objective avoidance behavior and subjective well-being after exposure, mediating the effect of the FSQ. In summary, the behavioral tasks are well suited to assess avoidance behavior in phobic participants and provide detailed insights into the process of avoidance.
Excitability-Independent Memory Allocation for Repeated Event
Authors: Cho Hye-Yeon , Lee Han-Sol , Jeong Yire , Han Junho , Yoo Miran , Han Jin-Hee
Published: April 2022

How memory is organized in cell ensembles when an event is repeated is not well-understood. Recently, we found that retraining 24 h after the initial fear conditioning (FC) event induces turnover of neurons in the lateral amygdala (LA) that encodes fear memory. Excitability-dependent competition between eligible neurons has been suggested as a rule that governs memory allocation. However, it remains undetermined whether excitability is also involved in the allocation of a repeated event. By increasing excitability in a subset of neurons in the LA before FC, we confirmed that these neurons preferentially participated in encoding fear memory as previously reported. These neurons, however, became unnecessary for memory recall after retraining 24 h following initial FC. Consistently, the initial memory-encoding neurons became less likely to be reactivated during recall. This reorganization in cell ensembles, however, was not induced and memory was co-allocated when retraining occurred 6 h after the initial FC. In 24-h retraining condition, artificially increasing excitability right before retraining failed to drive memory co-allocation. These results suggest a distinct memory allocation mechanism for repeated events distantly separated in time.
Robust Latent Multi-Source Adaptation for Encephalogram-Based Emotion Recognition
Authors: Tao Jianwen , Dan Yufang , Zhou Di , He Songsong
Published: April 2022

In practical encephalogram (EEG)-based machine learning, different subjects can be represented by many different EEG patterns, which would, in some extent, degrade the performance of extant subject-independent classifiers obtained from cross-subjects datasets. To this end, in this paper, we present a robust Latent Multi-source Adaptation (LMA) framework for cross-subject/dataset emotion recognition with EEG signals by uncovering multiple domain-invariant latent subspaces. Specifically, by jointly aligning the statistical and semantic distribution discrepancies between each source and target pair, multiple domain-invariant classifiers can be trained collaboratively in a unified framework. This framework can fully utilize the correlated knowledge among multiple sources with a novel low-rank regularization term. Comprehensive experiments on DEAP and SEED datasets demonstrate the superior or comparable performance of LMA with the state of the art in the EEG-based emotion recognition.
Corticofugal and Brainstem Functions Associated With Medial Olivocochlear Cholinergic Transmission
Authors: Munoz Felipe , Vicencio-Jimenez Sergio , Jorratt Pascal , Delano Paul H. , Terreros Gonzalo
Published: April 2022

Cholinergic transmission is essential for survival and reproduction, as it is involved in several physiological responses. In the auditory system, both ascending and descending auditory pathways are modulated by cholinergic transmission, affecting the perception of sounds. The auditory efferent system is a neuronal network comprised of several feedback loops, including corticofugal and brainstem pathways to the cochlear receptor. The auditory efferent system’s -final and mandatory synapses that connect the brain with the cochlear receptor- involve medial olivocochlear neurons and outer hair cells. A unique cholinergic transmission mediates these synapses through α9/α10 nicotinic receptors. To study this receptor, it was generated a strain of mice carrying a null mutation of the Chrna9 gene (α9-KO mice), lacking cholinergic transmission between medial olivocochlear neurons and outer hair cells, providing a unique opportunity to study the role of medial olivocochlear cholinergic transmission in auditory and cognitive functions. In this article, we review behavioral and physiological studies carried out to research auditory efferent function in the context of audition, cognition, and hearing impairments. Auditory studies have shown that hearing thresholds in the α9-KO mice are normal, while more complex auditory functions, such as frequency selectivity and sound localization, are altered. The corticofugal pathways have been studied in α9-KO mice using behavioral tasks, evidencing a reduced capacity to suppress auditory distractors during visual selective attention. Finally, we discuss the evolutionary role of the auditory efferent system detecting vocalizations in noise and its role in auditory disorders, such as the prevention of age-related hearing loss.
Generating Flight Illusions Using Galvanic Vestibular Stimulation in Virtual Reality Flight Simulations
Authors: Pradhan Gaurav N. , Galvan-Garza Raquel , Perez Alison M. , Bogle Jamie , Cevette Michael J.
Published: April 2022

Background

Vestibular flight illusions remain a significant source of concern for aviation training. Most fixed-based simulation training environments, including new virtual reality (VR) technology, lack the ability to recreate vestibular flight illusions as vestibular cues cannot be provided without stimulating the vestibular end organs. Galvanic vestibular stimulation (GVS) has long been used to create vestibular perception. The purpose of this study is to evaluate the ability of GVS to simulate common flight illusions by intentionally providing mismatched GVS during flight simulation scenarios in VR.

Methods

Nineteen participants performed two flight simulation tasks—take off and sustained turn—during two separate VR flight simulation sessions, with and without GVS (control). In the GVS session, specific multi-axis GVS stimulation (i.e., electric currents) was provided to induce approximate somatogravic and Coriolis illusions during the take-off and sustained turn tasks, respectively. The participants used the joystick to self-report their subjective motion perception. The angular joystick movement along the roll, yaw, and pitch axes was used to measure cumulative angular distance and peak angular velocity as continuous variables of motion perception across corresponding axes. Presence and Simulator Sickness Questionnaires were administered at the end of each session.

Results

The magnitude and variability of perceived somatogravic illusion during take-off task in the form of cumulative angular distance (p < 0.001) and peak velocity (p < 0.001) along the pitch-up axis among participants were significantly larger in the GVS session than in the NO GVS session. Similarly, during the sustained turn task, perceived Coriolis illusion in the form of cumulative angular distances (roll: p = 0.005, yaw: p = 0.015, pitch: p = 0.007) and peak velocities (roll: p = 0.003, yaw: p = 0.01, pitch: p = 0.007) across all three axes were significantly larger in the GVS session than in the NO GVS session. Subjective nausea was low overall, but significantly higher in the GVS session than in the NO GVS session (p = 0.026).

Discussion

Our findings demonstrated that intentionally mismatched GVS can significantly affect motion perception and create flight illusion perceptions during fixed-based VR flight simulation. This has the potential to enhance future training paradigms, providing pilots the ability to safely experience, identify, and learn to appropriately respond to flight illusions during ground training.
Effects of Oral Carotenoids on Oxidative Stress: A Systematic Review and Meta-Analysis of Studies in the Recent 20 Years
Authors: Zhuang Chengfei , Yuan Jinping , Du Yimei , Zeng Jing , Sun Yan , Wu Yan , Gao Xing-Hua , Chen Hong-Duo
Published: April 2022

Carotenoids protect organs, tissues, and cells from the damaging action of singlet oxygen, oxygen radicals, and lipid peroxides. This systematic review was sought to evaluate the influence of oral carotenoids on antioxidant/oxidative markers, blood carotenoids levels, and lipid/lipoprotein parameters in human subjects. A comprehensive review of relevant literature was conducted in PubMed, Web of Sciences, and the Cochrane library, from 2000 to December 2020. Randomized controlled trials, case-controlled trials, or controlled trials were identified. A total of eighteen trials were included, with the target populations being healthy subjects in 16 studies, athletes in 1 study, and pregnant women in 1 study. The meta-analysis results showed that carotenoids complex supplementation significantly increased the levels of antioxidative parameters ferric-reducing ability of plasma (FRAP) and oxygen radical absorbance capacity (ORAC) [standardized mean difference (SMD) = 0.468; 95% CI: 0.159–0.776, p = 0.003; SMD = 0.568; 95% CI: 0.190–0.947, p = 0.003] and decreased the blood triglyceride (TG) level (SMD = −0.410, 95% CI: −0.698 to −0.122, p = 0.005). Oral carotenoids supplement significantly increased the blood levels of β-carotene (SMD = 0.490, 95% CI: 0.123–0.858, p = 0.009), α-tocopherol (SMD = 0.752, 95%CI: 0.020–1.485, p = 0.044), and the intaking durations were 8 weeks. The levels of antioxidative enzymes and other lipid/lipoprotein parameters were not different between subjects receiving carotenoids and controls (p > 0.05). In conclusion, our systematic review showed that the carotenoids complex is beneficial for alleviating potential oxidative stress via interacting with free radicals or decreasing blood TG levels. The intaking duration of carotenoids should be 8 weeks to reach enough concentration for function.
Dietary Habits and Self-Reported Health Measures Among Norwegian Adults Adhering to Plant-Based Diets
Authors: Groufh-Jacobsen Synne , Bahr Bugge Annechen , Morseth Marianne Sandsmark , Pedersen Julia Tsuruta , Henjum Sigrun
Published: April 2022

Background

As plant-based diets is increasing, we aimed to investigate dietary habits, dietary motivation and self-reported health of Norwegian adults who adhere to different types of plant-based diets.

Methods

In 2020, 808 subjects (530 women and 278 men) participated in an online survey, including vegans (8%), lacto-ovo vegetarians (16%), pescatarians (23%), and flexitarians (53%).

Results

Half of the participants reported to consume fruits daily, three quarters consumed vegetables daily, and one quarter consumed whole grain products daily. Half of the participants reported weekly consumption of sweets and salted snacks, and 10% reported daily consumption of sugary drinks. Daily consumption of milk and dairy substitutes were reported by vegans (49%), lacto-ovo vegetarians (33%), pescatarians (32%), and flexitarians (25%). Daily consumption of meat replacement products was reported by vegans (3%), lacto-ovo vegetarians (5%), pescatarians (2%). Reported supplement use was highest among vegans (62%) and lowest among flexitarians (28%). Dietary motivations were driven by environmental (71%), health (64%), and animal welfare (49%) concerns, across all dietary groups. 75% reported their self-perceived health to be good or very good.

Conclusion

We found that Norwegian adults adhering to plant-based diets consumed less fruit, vegetables, and whole grain products, together with more sugar than recommended in the Norwegian dietary guidelines. The participants reported their self-perceived health to be good or very good.
Probiotic Effects of Bacillus licheniformis DSM5749 on Growth Performance and Intestinal Microecological Balance of Laying Hens
Authors: Pan Xue , Cai Yuanli , Kong Linglian , Xiao Chuanpi , Zhu Qidong , Song Zhigang
Published: April 2022

This study was conducted to investigate the effects of Bacillus licheniformis DSM5749 on the production performance and intestinal health in laying hens. A total of 32-week-old laying hens (Hyline Brown) were randomly assigned to two dietary groups (10 replicates of 27 laying hens), namely, basal diet and basal diet complemented with 200 g/t B. licheniformis (3.2 × 109 CFU/kg). The trial lasted for 8 weeks, and samples were collected at the last week. Results revealed that B. licheniformis DSM5749 significantly improved laying performance, including an increase in egg production rate and average daily egg yield, and a decrease in the feed-to-egg ratio during the entire 8-week experimental period (P < 0.05). B. licheniformis DSM5749 increased the levels of superoxide dismutase and glutathione peroxidase in the liver and decreased the IL-1 level in the serum (P < 0.05). In addition, the integrity of intestinal morphology (villus height, crypt depth, and villus height/crypt depth), tight junctions (ZO-1, Claudin-1, and Occludin), and lipase vitality in the intestine were potentiated by B. licheniformis DSM5749 in laying hens (P < 0.05). B. licheniformis DSM5749 decreased the Firmicutes/Bacteroidetes ratio (P < 0.05) in the cecum. Furthermore, B. licheniformis DSM5749 modulated the microbiota in the cecum of the laying hens, increased the relative abundance of beneficial bacteria (e.g., Prevotella) at the genus level and decreased the relative abundance of potential pathogens (e.g., Desulfovibrio). In conclusion, B. licheniformis DSM5749 can improve laying performance, promote intestinal health, affect the composition of cecal microorganisms, and regulate the intestinal micro-ecological balance, making B. licheniformis a good probiotic candidate for application in the laying hens industry.
Integrated Transcriptomic and Metabolomic Analyses Reveal the Mechanisms Underlying Anthocyanin Coloration and Aroma Formation in Purple Fennel
Authors: Zhang Yanjie , Zhao Qing , Feng Youwei , Dong Yuanhang , Zhang Tianjiao , Yang Qiu , Gu Huihui , Huang Jinyong , Li Yan
Published: April 2022

The color and aroma are the significant traits of vegetables and fruits, but the metabolic and molecular mechanisms underlying anthocyanin accumulation and aroma formation remain almost unknown in fennel (Anethum foeniculum L.), which is a crucial vegetable crop and grown widely for aromatic leaves and bulbs. Here, ten major anthocyanins identified and quantified by ultra-high performance liquid chromatography coupled with quadrupole Orbitrap high-resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS) were mainly responsible for the coloration of purple fennel leaf. With the application of GC-MS, it was found that the reduced volatile phenylpropanoids including isoeugenol, trans-isoeugenol, and apiol chiefly account for the characteristic aroma changes of the purple fennel. Moreover, the characteristic anthocyanin coloration and aroma formation in purple fennel were systematically studied with the integrated transcriptomics and metabolomics. The critical genes associated with the biosynthesis and regulation of anthocyanins and volatile phenylpropanoids were isolated and studied carefully in transiently transfected tobacco cells and transgenic tomato plants. Together with the results of UHPLC-Q-Orbitrap HRMS, RT-qPCR, and yeast two hybrid (Y2H), it is proved that the metabolic flux redirection of phenylpropanoid pathway primarily regulated by a functional MYB-bHLH-WD40 complex consisting of AfTT8, AfMYB7, and AfTTG1 accounts for the characteristic anthocyanin coloration and aroma formation in purple fennel leaf. The systematic understanding of the anthocyanin accumulation and aroma formation will assist in the improvement of fennel resource utilization and breeding.
Sparse Angle CBCT Reconstruction Based on Guided Image Filtering
Authors: Xu Siyuan , Yang Bo , Xu Congcong , Tian Jiawei , Liu Yan , Yin Lirong , Liu Shan , Zheng Wenfeng , Liu Chao
Published: April 2022

Cone-beam Computerized Tomography (CBCT) has the advantages of high ray utilization and detection efficiency, short scan time, high spatial and isotropic resolution. However, the X-rays emitted by CBCT examination are harmful to the human body, so reducing the radiation dose without damaging the reconstruction quality is the key to the reconstruction of CBCT. In this paper, we propose a sparse angle CBCT reconstruction algorithm based on Guided Image FilteringGIF, which combines the classic Simultaneous Algebra Reconstruction Technique(SART) and the Total p-Variation (TpV) minimization. Due to the good edge-preserving ability of SART and noise suppression ability of TpV minimization, the proposed method can suppress noise and artifacts while preserving edge and texture information in reconstructed images. Experimental results based on simulated and real-measured CBCT datasets show the advantages of the proposed method.
Extracellular DNA Traps: Origin, Function and Implications for Anti-Cancer Therapies
Authors: Mamtimin Medina , Pinarci Akif , Han Chao , Braun Attila , Anders Hans-Joachim , Gudermann Thomas , Mammadova-Bach Elmina
Published: April 2022

Extracellular DNA may serve as marker in liquid biopsies to determine individual diagnosis and prognosis in cancer patients. Cell death or active release from various cell types, including immune cells can result in the release of DNA into the extracellular milieu. Neutrophils are important components of the innate immune system, controlling pathogens through phagocytosis and/or the release of neutrophil extracellular traps (NETs). NETs also promote tumor progression and metastasis, by modulating angiogenesis, anti-tumor immunity, blood clotting and inflammation and providing a supportive niche for metastasizing cancer cells. Besides neutrophils, other immune cells such as eosinophils, dendritic cells, monocytes/macrophages, mast cells, basophils and lymphocytes can also form extracellular traps (ETs) during cancer progression, indicating possible multiple origins of extracellular DNA in cancer. In this review, we summarize the pathomechanisms of ET formation generated by different cell types, and analyze these processes in the context of cancer. We also critically discuss potential ET-inhibiting agents, which may open new therapeutic strategies for cancer prevention and treatment.
Effects of Proinflammatory Cytokines on Lacrimal Gland Myoepithelial Cells Contraction
Authors: Garriz Angela , Morokuma Junji , Bowman Maytal , Pagni Sarah , Zoukhri Driss
Published: April 2022

In the lacrimal gland, myoepithelial cells (MEC) express muscle contractile proteins such as alpha smooth muscle actin (SMA) and calponin and therefore can contract to help expel lacrimal fluid. In a previous study, we demonstrated that lacrimal gland MEC express the oxytocin receptor (OXTR) and they contract under oxytocin (OXT) stimulation. Using NOD and MRL/lpr mice (animal models of Sjogren’s syndrome), we reported a decrease in SMA and calponin protein levels plus a decline in acini contraction after stimulation with OXT. It is known that proinflammatory cytokines, such as interleukin-1β (IL-1β), tumor necrosis factor alpha (TNF-α) or interferon gamma (IFN-γ), can affect OXTR expression and signaling capacity and inhibit MEC contraction. The aim of the current study was to investigate if proinflammatory cytokines are implicated in the loss of MEC contractile ability. Thus, lacrimal gland MEC from SMA-GFP transgenic mice were treated with IL-1β (10 ng/ml) for a total of 7 days. At days 0, 2, 4 and 7, GFP intensity, cell size/area, contractile proteins amounts and MEC contraction were assessed. At day 0, control and treated cells showed no differences in GFP intensity and cell size. GFP intensity started to decrease in treated MEC at day 2 (20%; p=0.02), continuing after day 4 (25%; p=0.007) and 7 (30%; p=0.0001). Mean cell area was also reduced at day 2 (34%; p=0.0005), and after 4 (51%; p<0.0001) and 7 days (30%; p=0.0015). The contraction assay at day 2 showed a 70% decrease of contraction in treated MEC (p<0.0001), 73% (p<0.0001) at day 4 and 82% (p=0.0015) at day 7 when compared to control. Levels of contractile proteins were measured on day 7 showing a decrease in SMA and calponin amount in treated MEC compared with the control group (around 30%; p=0.0016 and p=0.0206; respectively). Similar results were observed when TNF-α and IFN-γ were added along with IL-1β. Taken together the present data and those from our previous studies with Sjogren’s syndrome mouse models, they strongly suggest that proinflammatory cytokines affect lacrimal gland MEC contractile ability that may account for the reduced tear secretion associated with Sjogren’s syndrome dry eye disease.
Number of Episodes Can Be Used as a Disease Activity Measure in Familial Mediterranean Fever
Authors: Piskin David , Arici Zehra Serap , Konukbay Dilek , Romano Micol , Makay Balahan , Ayaz Nuray , Bilginer Yelda , Berard Roberta A. , Poyrazoglu Hakan , Kasapcopur Ozgur , Laxer Ronald M. , Speechley Kathy , Demirkaya Erkan
Published: April 2022

Objective

To evaluate the number of episodes in the past 12 months as an indicator of the overall disease activity status in Familial Mediterranean fever (FMF).

Methods

In this cross-sectional study, patients were recruited from tertiary pediatric hospitals. Demographic data, main clinical symptoms of the episodes, treatment modalities, and genetic mutations were recorded. The patients were grouped as no episodes (Group 1), 1–4 episodes (Group 2), and more than 4 episodes (Group 3) according to the number of episodes in the past 12 months. The Pediatric Quality Life Inventory (PedsQL), the Children’s Depression Inventory (CDI), and the Wong-Baker FACES Pain Rating Scale (FACES) scores were compared between groups. Concurrent validity between the number of episodes and the patient-reported outcome measures (PROMs) was assessed using Spearman’s rank correlation coefficient (ρ).

Results

A total of 239 patients were included. There were 74 patients (31%) in Group 1, 99 (41.4%) in Group 2, and 66 (27.6%) in Group 3. Groups were similar according to age, age at diagnosis, gender, consanguinity, family history, history of amyloidosis, clinical symptoms, and in terms of allele frequency (p > 0.05). According to PROMs completed by parents, moderate correlations were found between the number of episodes and the PedsQL score (ρ = −0.48; 95% CI = −0.58 to −0.35, p < 0.001) and between the number of episodes and the Wong-Baker FACES score (ρ = 0.47, 95% CI = 0.35–0.57, p < 0.001).

Conclusion

The number of episodes was positively and moderately correlated with patient- and parent-reported outcomes in our cohort. The number of episodes in patients with FMF can be used as a single measure to assess disease activity.
Machine Learning-Based Prediction Method for Tremors Induced by Tacrolimus in the Treatment of Nephrotic Syndrome
Authors: Shao Bing , Qu Youyang , Zhang Wei , Zhan Haihe , Li Zerong , Han Xingyu , Ma Mengchao , Du Zhimin
Published: April 2022

Tremors have been reported even with a low dose of tacrolimus in patients with nephrotic syndrome and are responsible for hampering the day-to-day work of young active patients with nephrotic syndrome. This study proposes a neural network model based on seven variables to predict the development of tremors following tacrolimus. The sensitivity and specificity of this algorithm are high. A total of 252 patients were included in this study, out of which 39 (15.5%) experienced tremors, 181 patients (including 32 patients who experienced tremors) were randomly assigned to a training dataset, and the remaining were assigned to an external validation set. We used a recursive feature elimination algorithm to train the training dataset, in turn, through 10-fold cross-validation. The classification performance of the classifer was then used as the evaluation criterion for these subsets to find the subset of optimal features. A neural network was used as a classification algorithm to accurately predict tremors using the subset of optimal features. This model was subsequently tested in the validation dataset. The subset of optimal features contained seven variables (creatinine, D-dimer, total protein, calcium ion, platelet distribution width, serum kalium, and fibrinogen), and the highest accuracy obtained was 0.8288. The neural network model based on these seven variables obtained an area under the curve (AUC) value of 0.9726, an accuracy of 0.9345, a sensitivity of 0.9712, and a specificity of 0.7586 in the training set. Meanwhile, the external validation achieved an accuracy of 0.8214, a sensitivity of 0.8378, and a specificity of 0.7000 in the validation dataset. This model was capable of predicting tremors caused by tacrolimus with an excellent degree of accuracy, which can be beneficial in the treatment of nephrotic syndrome patients.
Astragaloside IV Alleviates Intestinal Barrier Dysfunction via the AKT-GSK3β-β-Catenin Pathway in Peritoneal Dialysis
Authors: He Jiaqi , Wang Mengling , Yang Licai , Xin Hong , Bian Fan , Jiang Gengru , Zhang Xuemei
Published: April 2022

Background and aims: Long-term peritoneal dialysis (PD) causes intestinal dysfunction, including constipation, diarrhea, or enteric peritonitis. However, the etiology and pathogenesis of these complications are still unclear and there are no specific drugs available in the clinic. This study aims to determine whether Astragaloside IV (AS IV) has therapeutic value on PD-induced intestinal epithelial barrier dysfunction in vivo and in vitro.

Methods: We established two different long-term PD treatment mice models by intraperitoneally injecting 4.25% dextrose-containing peritoneal dialysis fluid (PDF) in uremia mice and normal mice, which were served as controls. In addition, PDF was applied to T84 cells in vitro. The therapeutic effects of AS IV on PD-induced intestinal dysfunction were then examined by histopathological staining, transmission electron microscopy, western blotting, and reverse transcription polymerase chain reaction. The protein levels of protein kinase B (AKT), glycogen synthase kinase 3β (GSK-3β) and β-catenin were examined after administration of AS IV.

Results: In the present study, AS IV maintained the intestinal crypt, microvilli and desmosome structures in an orderly arrangement and improved intestinal epithelial permeability with the up-regulation of tight junction proteins in vivo. Furthermore, AS IV protected T84 cells from PD-induced damage by improving cell viability, promoting wound healing, and increasing the expression of tight junction proteins. Additionally, AS IV treatment significantly increased the levels of phosphorylation of AKT, inhibited the activity GSK-3β, and ultimately resulted in the nuclear translocation and accumulation of β-catenin.

Conclusion: These findings provide novel insight into the AS IV-mediated protection of the intestinal epithelial barrier from damage via the AKT-GSK3β-β-catenin signal axis during peritoneal dialysis.
Baohuoside I Inhibits Osteoclastogenesis and Protects Against Ovariectomy-Induced Bone Loss
Authors: Ma Min , Fan Ao-yuan , Liu Zheng , Yang Li-qing , Huang Jun-ming , Pang Zhi-ying , Yin Feng
Published: April 2022

Bone-resorbing osteoclasts are essential for skeletal remodelling, and the hyperactive formation and function of osteoclasts are common in bone metabolic diseases, especially postmenopausal osteoporosis. Therefore, regulating the osteoclast differentiation is a major therapeutic target in osteoporosis treatment. Icariin has shown potential osteoprotective effects. However, existing studies have reported limited bioavailability of icariin, and the material basis of icariin for anti-osteoporosis is attributed to its metabolites in the body. Here, we compared the effects of icariin and its metabolites (icariside I, baohuoside I, and icaritin) on osteoclastogenesis by high-content screening followed by TRAP staining and identified baohuoside I (BS) with an optimal effect. Then, we evaluated the effects of BS on osteoclast differentiation and bone resorptive activity in both in vivo and in vitro experiments. In an in vitro study, BS inhibited osteoclast formation and bone resorption function in a dose-dependent manner, and the elevated osteoclastic-related genes induced by RANKL, such as NFATc1, cathepsin K, RANK, and TRAP, were also attenuated following BS treatment. In an in vivo study, OVX-induced bone loss could be prevented by BS through interrupting the osteoclast formation and activity in mice. Furthermore, mechanistic investigation demonstrated that BS inhibited osteoclast differentiation by ameliorating the activation of the MAPK and NF-kB pathways and reducing the expression of uPAR. Our study demonstrated that baohuoside I could inhibit osteoclast differentiation and protect bone loss following ovariectomy.
Immunophenotyping of Acute Inflammatory Exacerbations of Lung Injury Driven by Mutant Surfactant Protein-C: A Role for Inflammatory Eosinophils
Authors: Nguyen Jacklyn , Armstrong Brittnie S. , Cowman Sophie , Tomer Yaniv , Veerabhadraiah Shivakumar R. , Beers Michael F. , Venosa Alessandro
Published: April 2022

Acute inflammatory exacerbations (AIEs) represent immune-driven deteriorations of many chronic lung conditions, including COPD, asthma, and pulmonary fibrosis (PF). The first line of therapy is represented by broad-spectrum immunomodulation. Among the several inflammatory populations mobilizing during AIEs, eosinophils have been identified as promising indicators of an active inflammatory exacerbation. To better study the eosinophil-parenchymal crosstalk during AIE-PF, this work leverages a clinically relevant model of inflammatory exacerbations triggered by inducible expression of a mutation in the alveolar epithelial type 2 cell Surfactant Protein-C gene [SP-CI73T]. Unbiased single-cell sequencing analysis of controls and SP-CI73T mutants at a time coordinated with peak eosinophilia (14 days) defined heightened inflammatory activation, chemotaxis, and survival signaling (IL-6, IL-4/13, STAT3, Glucocorticoid Receptor, mTOR, and MYC) in eosinophils. To study the impact of eosinophils in inflammatory exacerbations, the SP-CI73T line was crossed with eosinophil lineage deficient mice (GATA1Δdbl) to produce the SP-CI73TGATA1KO line. Time course analysis (7–42 days) demonstrated improved lung histology, survival, and reduced inflammation in SP-CI73TGATA1KO cohorts. Spectral flow cytometry of tissue digests confirmed eosinophil depletion in GATA1KO mice and the absence of a compensatory shift in neutrophils and immature monocyte recruitment. Eosinophil deletion resulted in progressive monocyte-derived macrophage accumulation (14 days post-injury), combined with declines in CD3+CD4+ lymphocyte and B220+ B cell abundance. Histochemical analysis revealed atypical inflammatory cell activation in SP-CI73TGATA1KO mice, with reduced numbers of Arg-1+ and iNOS+ cells, but increases in tgfb1 mRNA expression in bronchoalveolar lavage cells and tissue. Dexamethasone treatment (1 mg/kg daily, i.p.) was utilized to investigate corticosteroid efficacy in highly eosinophilic exacerbations induced by mutant SP-CI73T. Dexamethasone successfully reduced total and eosinophil (CD11b+SigF+CD11c−) counts at 14 days and was linked to reduced evidence of structural damage and perivascular infiltrate. Together, these results illustrate the deleterious role of eosinophils in inflammatory events preceding lung fibrosis and demonstrate the efficacy of corticosteroid treatment in highly eosinophilic exacerbations induced by mutant SP-CI73T.
Discovery of METTL3 Small Molecule Inhibitors by Virtual Screening of Natural Products
Authors: Du Yue , Yuan Yongliang , Xu Le , Zhao Fang , Wang Wenbin , Xu Yiping , Tian Xin
Published: April 2022

N6-Methyladenosine (m6A) is the most prevalent mRNA modification in mammalian cells that is mainly catalyzed by the methyltransferase complex of methyltransferase-like 3 and methyltransferase-like 14 (METTL3-METTL14). Many lines of evidence suggest that METTL3 plays important roles in several diseases such as cancers and viral infection. In the present study, 1,042 natural products from commercially available sources were chosen to establish a screening library, and docking-based high-throughput screening was performed to discover potential METTL3 inhibitors. The selected compounds were then further validated by an in vitro methyltransferase inhibition assay in which m6A content was determined by LC-MS/MS. A cellular assay of the inhibition of m6A methylation was performed to determine the METTL3 inhibitory activity of the selected compound. CCK-8 assay was applied to evaluate the effects of the selected compound on tumor cell viability. Additionally, binding mode analysis, molecular dynamics (MD) simulation, and binding free energy analysis were performed to study the process and characteristics of inhibitor binding. Finally, quercetin was identified as a METTL3 inhibitor with an IC50 value of 2.73 μM. The cellular assay of m6A methylation inhibition showed that quercetin decreased m6A level in a dose-dependent manner in MIA PaCa-2 pancreatic cancer cells. CCK-8 assay showed quercetin efficiently inhibited the proliferation of MIA PaCa-2 and Huh7 tumor cells, with IC50 values 73.51 ± 11.22 μM and 99.97 ± 7.03 μM, respectively. Molecular docking studies revealed that quercetin filled the pocket of the adenosine moiety of SAM but not the pocket of the SAM methionine in the METTL3 protein, and hydrogen bonds, hydrophobic interactions, and pi-stacking were formed. The values of the root mean square deviation (RMSD), the root mean square fluctuations (RMSF), and binding free energy suggested that quercetin can efficiently bind to the pocket of the METTL3 protein and form a stable protein-ligand complex. The present study is the first to identify METTL3 inhibitors from natural products, thus providing a basis for subsequent research and facilitating the development of METTL3-targeting drugs for diseases.