bluuebunny/crossref_metadata_2025 huggingface dataset:
Microbial Community Affects Daqu Quality and the Production of Ethanol and Flavor Compounds in Baijiu Fermentation
Authors: Han Pei-Jie , Luo Lu-Jun , Han Ying , Song Liang , Zhen Pan , Han Da-Yong , Wei Yu-Hua , Zhou Xin , Wen Zhang , Qiu Jun-Zhi , Bai Feng-Yan
Published: August 2023

Daqu is a traditional starter for Baijiu fermentation and is produced by spontaneous fermentation of ground and moistened barley or wheat. The quality of Daqu is traditionally evaluated based on physicochemical and subjective sensory parameters without microbiological analysis. Here, we compared the physicochemical characteristics of qualified (QD) and inferior (ID) Daqu, their microbial communities based on plate counting and PacBio SMRT sequencing of rRNA gene libraries, and their impacts on Baijiu fermentation. The results showed that the glucoamylase and α-amylase activities of QD were significantly higher than those of ID. The counts of yeasts and relative abundances of functional microbes, especially the amylolytic bacterium Bacillus licheniformis and fungi Saccharomycopsis fibuligera and Lichtheimia ramosa, were significantly higher in QD than in ID. The laboratory-scale Baijiu fermentation tests showed that the relative abundances of the amylolytic microbes were higher in the QD than the ID fermentation set, resulting in more efficient fermentation, as indicated by more weight loss and higher moisture content in the former. Consequently, more glycerol, acetic acid, ethanol, and other volatile compounds were produced in the QD than in the ID fermentation set. The results suggest that Daqu quality is determined by, and can be evaluated based on, its microbial community.
Slope Failure and Landslide Detection in Huangdao District of Qingdao City Based on an Improved Faster R-CNN Model
Authors: Guan Yong , Yu Lili , Hao Shengyou , Li Linsen , Zhang Xiaotong , Hao Ming
Published: August 2023

To reduce the significant losses caused by slope failures and landslides, it is of great significance to detect and predict these disasters scientifically. This study focused on Huangdao District of Qingdao City in Shandong Province, using the improved Faster R-CNN network to detect slope failures and landslides. This study introduced a multi-scale feature enhancement module into the Faster R-CNN model. The module enhances the network’s perception of different scales of slope failures and landslides by deeply fusing high-resolution weak semantic features with low-resolution strong semantic features. Our experiments show that the improved Faster R-CNN model outperformed the traditional version, and that ResNet50 performed better than VGG16 with an AP value of 90.68%, F1 value of 0.94, recall value of 90.68%, and precision value of 98.17%. While the targets predicted by VGG16 were more dispersed and the false detection rate was higher than that of ResNet50, VGG16 was shown to have an advantage in predicting small-scale slope failures and landslides. The trained Faster R-CNN network model detected geological hazards of slope failure and landslide in Huangdao District, missing only two landslides, thereby demonstrating high detection accuracy. This method can provide an effective technical means for slope failures and landslides target detection and has practical implications.
A Universal Graphical Solution to Calculating Seepage in Excavation of Anisotropic Soils and Non-Limited Scenarios
Authors: Navarro Carrasco Salvador , Jiménez-Valera José Antonio , Alhama Ivan
Published: August 2023

The interaction between groundwater and civil engineering works is a key aspect in geotechnical design. In the case of excavations confined in sheet pile walls, steel sheeting, diaphragm walls, cut-off walls, or cofferdams, this design requires the estimation, among other soil mechanics properties, of the groundwater flowing into the excavation (seepage) caused by piezometry depletion. Numerical methods, graphical solutions, and analytical procedures are the methodologies traditionally used to solve this issue, solutions of which require an understanding of basic soil mechanical properties, hydraulic conditions and structure geometry. In this work, the discriminated non-dimensionalization technique is applied to obtain, for the first time, the dimensionless groups that govern the seepage, in anisotropic conditions, in large-scale scenarios where groundwater flow is not conditioned by impervious bedrock or the length of the back of the wall: π1=ab,π2=kxb2kyc2 and, π3=T/b. Numerical simulations are carried out to check the validity of dimensionless groups and to develop three sets of type curves that relate to these groups. Once the physical and geometrical data are known, the seepage (Q), the characteristic depth (T*) and the characteristic horizontal extension (L*) can be directly and easily calculated from these abacuses. The influence of anisotropy on the characteristic lengths is also addressed.
Identification of Sociodemographic and Clinical Factors Influencing the Feeling of Stigmatization in People with Type 1 Diabetes
Authors: Sińska Beata I. , Kucharska Alicja , Panczyk Mariusz , Matejko Bartłomiej , Traczyk Iwona , Harton Anna , Jaworski Mariusz
Published: August 2023

Background: A large percentage of people with type 1 diabetes experience stigma, which may directly affect diabetes management. Moreover, it may adversely influence the acceptance of the disease and, thus, the treatment process, including compliance with medical and dietary recommendations. Therefore, it is important to seek adequate forms of counteracting the phenomenon of stigmatization. Thus, the aim of the study was to determine the factors influencing the level of perceived stigma by T1D patients, with particular emphasis on sociodemographic factors (including sex, place of residence, and education) and clinical factors related to the course of the disease. Methods: An observational cross-sectional online questionnaire was conducted in a group of 339 people with T1D. The link to the questionnaire was shared via social media. The DSAS-1 questionnaire translated into Polish was used as the research tool. Results: A moderate level of stigmatization was found (49.78 ± 14.54 points). It was significantly lower in people living in small towns compared to rural residents (ß = −0.121, p = 0.038), lower in people in relationships compared to those who are single (ß = −0.175, p = 0.001), in people diagnosed with T1D at an older age (ß = −0.107, p = 0.048), and in those who rated their financial situation as very good vs. bad (ß = −0.314, p < 0.001). It was also found that the level of stigma significantly decreased with age (ß = −0.181, p = 0.001). In addition, a significantly higher stigma perception was uncovered in the group of people with HbA1C > 7% than in the group ≤ 7% (ß = 0.118, p = 0.030). Conclusions: Due to the occurrence of stigma among people with T1D, which can directly affect the management of diabetes, effective and comprehensive efforts should be made to provide support to people with diabetes. It is also necessary to raise awareness among the general public and disseminate knowledge about diabetes, which can reduce stigmatization as a result. Anti-stigma messages should be included in the prevention programs about the potential side effects and risks associated with mistreating people with T1D.
Efficacy of Bendamustine, Pomalidomide, and Dexamethasone (BPD) Regimen in Relapsed/Refractory Extramedullary Myeloma: A Retrospective Single-Centre Study, Real-Life Experience
Authors: Açar İbrahim Halil , Güvenç Birol
Published: August 2023

Background and Objectives: Relapsed/refractory extramedullary myeloma (RREMM) is an uncommon and aggressive subtype of multiple myeloma defined by plasma cell proliferation outside the bone marrow. Therapeutic options for RREMM are limited, and the prognosis is generally unfavorable. This research aimed to assess the effectiveness of the bendamustine, pomalidomide, and dexamethasone (BPD) regimen in patients with RREMM. Material and Methods: We carried out a retrospective investigation of 11 RREMM patients who underwent BPD treatment. The primary endpoint was progression-free survival. The secondary endpoints of the study were two-year survival and overall response rate (ORR). We analyzed the sociodemographic and clinical features of the patients. Results: The average age of the patients was 62 years. They had a median of four prior treatment lines, and eight patients had previously received autologous stem-cell transplantation. After eight BPD treatment cycles, the ORR stood at 54%, with one very good partial response (VGPR), five partial responses (PR), three progressive diseases (PD), and two stable diseases (SD). The median follow-up was 15 months, with a two-year PFS rate of 71.3% and a two-year survival rate of 81.8%. Conclusions: The BPD regimen demonstrated promising effectiveness in RREMM patients, yielding favorable ORR and survival rates. To corroborate these findings and explore additional treatment alternatives for this patient group, larger prospective studies are required.
Combined Effect of Leisure-Time Physical Activity and Sedentary Behavior on Abdominal Obesity in ELSA-Brasil Participants
Authors: de Araújo Maiara do Espírito Santo Cerqueira , da Conceição Chagas de Almeida Maria , Matos Sheila Maria Alvim , de Jesus Mendes da Fonseca Maria , Pitanga Cristiano Penas Seara , Pitanga Francisco José Gondim
Published: August 2023

Introduction: Abdominal obesity is a pattern of obesity that has been considered a public health problem. Physical activity is considered an important factor for the prevention of abdominal obesity. Increased time in sedentary behavior has been associated with negative health outcomes, including abdominal obesity. Objective: The aim of this study was to investigate which combination of leisure-time physical activity and sedentary behavior contributes most to the prevention of abdominal obesity in adults participating in ELSA-Brasil (Longitudinal Study of Adult Health). Methods: The study was cross-sectional and participants from the first follow-up of the ELSA-Brasil cohort (2012–2014) were analyzed. The independent variables were physical activity, assessed by IPAQ, and sedentary behavior, assessed by a standard questionnaire applied in ELSA-Brasil; the dependent variable was abdominal obesity, determined by waist circumference. The covariates analyzed were the following: age, education, binge drinking, smoking and menopause. The associations between the dependent variable and the independent variables were analyzed using logistic regression. The odds ratio with 95 CI% was estimated. Results: For men, the combinations were more significant when they were more physically active and spent less time on the sedentary behaviors analyzed, on both a weekday and a weekend day. For menopausal women, both younger and older, all associations of the combinations between sufficient leisure-time physical activity and little time spent in sedentary behaviors contributed to the prevention of abdominal obesity. In non-menopausal women, positive associations were observed in almost all combinations between leisure-time physical activity and sedentary behaviors, with some results that were not statistically significant among younger women. Conclusions: Our results showed that being sufficiently active and reducing the time spent in sedentary behavior was the combination that contributed the most to the prevention of abdominal obesity, both in men and women.
Longitudinal Examination of Which Married Young Women Use Contraception to Delay a First Pregnancy in Bihar and Uttar Pradesh, India
Authors: Speizer Ilene S. , Zavier A. J. Francis , Calhoun Lisa M. , Hainsworth Gwyn , Guilkey David K.
Published: August 2023

Early marriage and childbearing put young women and their babies at risk of poor health and well-being. This study uses two rounds of longitudinal data from young women ages 15–19 in 2015–2016 and followed in 2018–2019 to determine factors associated with contraceptive use before a first pregnancy among young, married women in Bihar and Uttar Pradesh, India. Discrete time hazard models were used to analyze time to first use starting from the month of marriage. Overall, use of contraception prior to a first pregnancy was low in this sample (between 12 to 20% used before a first pregnancy). Young women who reported that someone discussed the importance of delaying a first birth at the time of marriage were significantly more likely to have used a method of family planning (FP) before a first pregnancy than those who did not receive this information. Further, women who discussed FP with their husband before a first pregnancy were more likely to use contraception. Finally, among recently married young women, those who experienced pressure to have a child were less likely to use before a first pregnancy. As young women recognize the advantages of delaying a first birth and adopt FP to meet their needs, social norms around early childbearing will slowly adjust and early use to delay a first pregnancy will become more normative.
Defending Others Online: The Influence of Observing Formal and Informal Social Control on One’s Willingness to Defend Cyberhate Victims
Authors: Costello Matthew , Hawdon James , Reichelmann Ashley V. , Oksanen Atte , Blaya Catherine , Llorent Vicente J. , Räsänen Pekka , Zych Izabela
Published: August 2023

This paper examines factors correlated with online self-help—an informal form of social control vis-à-vis intervention—upon witnessing a cyberhate attack. Using online surveys from 18- to 26-year-old respondents in the United States, we explore the roles of various types of online and offline formal and informal social control mechanisms on the enactment of self-help through the use of descriptive statistics and ordinal logistic regression. The results of the multivariate analyses indicate that online collective efficacy is positively related to self-help, as is having close ties to individuals and groups offline and online. Formal online social control, however, is not significantly related to engaging in self-help. Other findings demonstrate that personal encounters with cyberhate affect the likelihood that an individual will intervene when witnessing an attack, and that individuals with high levels of empathy are more likely to intervene to assist others. This work indicates that pro-social online behavior is contagious and can potentially foster online spaces in which harmful behaviors, such as propagating cyberhate, are not condoned.
Smartphone Distraction: Italian Validation of the Smartphone Distraction Scale (SDS)
Authors: Mascia Maria Lidia , Agus Mirian , Tomczyk Łukasz , Bonfiglio Natale Salvatore , Bellini Diego , Penna Maria Pietronilla
Published: August 2023

This work aimed to validate the use of the Smartphone Distraction Scale (SDS) in Italy. The SDS was devised to assess distraction related to smartphone use in adult populations. A cross-sectional study was conducted among n = 609 adults (females = 76.4%; mean age = 30.26; SD age = 9.90). An assessment of the factorial structure of the Italian version was carried out using exploratory factor analysis (EFA) and confirmatory factor analysis (CFA). The four factors identified by Throuvala and colleagues were confirmed (i.e., attention impulsiveness, online vigilance, multitasking and emotion regulation). Reliability was assessed using Cronbach’s alpha coefficient (0.703–0.889). The scale’s scores showed significant linear correlations with validated instruments, including the Mobile Phone Problematic Use Scale (MPPUS)and the Cognitive Failures Questionnaire (CFQ). A multivariate analysis of variance showed significant differences in the means among participants belonging to different age groups (born before 1995 vs. born after 1996). In summary, the good psychometric properties observed led us to assume that this instrument can be applied and used in Italian studies to assess the cognitive dimension of distraction related to the use of smartphones.
The Spatial Mechanism and Predication of Rural Tourism Development in China: A Random Forest Regression Analysis
Authors: Du Xishihui , Wang Zhaoguo , Wang Yan
Published: August 2023

Rural tourism has long been recognized as a significant strategy for promoting rural revitalization in China. Excessive development has had a number of negative consequences for rural tourism. As a result, there is a growing need to optimize the developmental framework of rural tourism in order to ensure its sustainable growth. This study focuses on key tourism villages and employs geostatistical analysis and the random forest methodology to elucidate the spatial mechanisms underlying rural tourism and identify potential areas for its development in China. The research findings reveal several important insights: (1) Key tourism villages exhibit a concentrated spatial distribution, characterized by pronounced regional disparities. (2) The intrinsic characteristics of rural areas and the conditions conducive to tourism development play pivotal roles in shaping rural tourism. Notably, cultural resources, tourism resources, rural accessibility, and tourism potential are identified as the primary influential factors. (3) Predictive modeling using random forest analysis indicates that densely populated areas in the eastern region retain the highest level of suitability for rural tourism. In contrast, the development of rural tourism in western and border regions encounters certain constraints. Additionally, the northern region encompasses larger expanses with high suitability, whereas the southern region is generally moderate. This comprehensive nationwide investigation provides valuable insights into the key aspects of rural tourism development and offers practical guidance for achieving sustainable rural tourism practices in China.
Analysis of Correlation between Anthropization Phenomena and Landscape Values of the Territory: A GIS Framework Based on Spatial Statistics
Authors: García-Ayllón Salvador , Martínez Gloria
Published: August 2023

The evaluation of anthropogenic impacts on the landscape is an issue that has traditionally been carried out from a descriptive or at least somewhat qualitative perspective. However, in recent years, the technological improvements provided by geographic information systems (GIS) and spatial statistics have led to more objective methodological frameworks for analysis based on quantitative approaches. This study proposes an innovative methodological framework for the evaluation of landscape impacts of the usual anthropization phenomena, using a retrospective spatiotemporal analysis based on geostatistical indicators. Various territorial indices have been used to assess the spatiotemporal evolution of fragmentation of the built-up urban fabric, the construction of roads or linear communication works and the changes in land use. These phenomena have been statistically correlated with objective indicators of the landscape’s intrinsic value. The analysis of said spatial statistical correlation has been applied to three different but neighboring environments in the region of Murcia, located in the southeast of Mediterranean Spain, providing interesting results on the objective impact of each of these phenomena on the landscape and depending on the boundary conditions.
A Lighting Consistency Technique for Outdoor Augmented Reality Systems Based on Multi-Source Geo-Information
Authors: Zhu Kunpeng , Liu Shuo , Sun Weichao , Yuan Yixin , Wu Yuang
Published: August 2023

Achieving seamless integration between virtual objects and real scenes has always been an important issue in augmented reality (AR) research. To achieve this, it is necessary to provide virtual objects with real-time and accurate lighting conditions from a real scene. Therefore, the purpose of this study is to realize lighting consistency rendering for real-time AR systems in outdoor environments, aiming to enhance the user’s sense of immersion. In this paper, we propose a lighting consistency technique for real-time AR systems in outdoor environments based on multi-source geographical information (MGI). Specifically, we introduce MGI into the study of lighting consistency and construct a comprehensive database to store and manage the acquired MGI data. Based on this, we proposed a sky radiance model driven using the MGI. Finally, we utilized the sky radiance model along with light sensor data to render the virtual objects in outdoor scenes. The experimental results show that the shadow angular error is reduced to 5.2°, and the system frame rate is increased to 94.26. This means that our method achieves a high level of realism in the fusion of virtual objects and real scenes while ensuring a high frame rate in the system. With this technology, users can conveniently and extensively realize the lighting consistency rendering of real-time AR systems in outdoor scenes using mobile devices.
Significance of Th17 and Treg in Treatment Efficacy and Outcome in Pediatric Acute Lymphoblastic Leukemia
Authors: Krętowska-Grunwald Anna , Sawicka-Żukowska Małgorzata , Kowalska Małgorzata , Basaj Aleksandra , Krawczuk-Rybak Maryna , Moniuszko Marcin , Grubczak Kamil
Published: August 2023

Acute lymphoblastic leukemia represents a malignant proliferation of lymphoid cells blocked at an early stage of cell differentiation. It is the most common cancer occurring in children. Despite favorable prognosis, the survival rate of patients with poor treatment response or relapse remains dismal. The interaction between leukemic cells and the tumor immune microenvironment is pivotal in mediating tumor progression. In this study we evaluated associations between Treg and Th17 lymphocytes and the clinical presentation of ALL pediatric patients to validate their value in monitoring treatment outcome. The peripheral blood and bone marrow aspirates from 35 pediatric patients with ALL and 48 healthy control subjects were selected for the experiment. We demonstrated the numbers of Th17 lymphocytes and Tregs were increased in the bone marrow of ALL patients at the moment of diagnosis compared to the healthy control group, with the latter significantly decreasing during the course of ALL treatment. Patients with lower Th17 were found to demonstrate higher risk of blasts prevalence in bone marrow at day 33. ALL patients with lower WBC demonstrated higher frequency of Tregs. In summary, we identified a significant role of Th17 and Treg lymphocytes in ALL of pediatric patients and their contribution to disease-related parameters.
Genome-Wide Identification and Characterization Analysis of WUSCHEL-Related Homeobox Family in Melon (Cucumis melo L.)
Authors: Tang Lingli , He Yuhua , Liu Bin , Xu Yongyang , Zhao Guangwei
Published: August 2023

WUSCHEL-related homeobox (WOX) proteins are very important in controlling plant development and stress responses. However, the WOX family members and their role in response to abiotic stresses are largely unknown in melon (Cucumis melo L.). In this study, 11 WOX (CmWOX) transcript factors with conserved WUS and homeobox motif were identified and characterized, and subdivided into modern clade, ancient clade and intermediate clade based on bioinformatic and phylogenetic analysis. Evolutionary analysis revealed that the CmWOX family showed protein variations in Arabidopsis, tomato, cucumber, melon and rice. Alignment of protein sequences uncovered that all CmWOXs had the typical homeodomain, which consisted of conserved amino acids. Cis-element analysis showed that CmWOX genes may response to abiotic stress. RNA-seq and qRT-PCR results further revealed that the expression of partially CmWOX genes are associated with cold and drought. CmWOX13a and CmWOX13b were constitutively expressed under abiotic stresses, CmWOX4 may play a role in abiotic processes during plant development. Taken together, this study offers new perspectives on the CmWOX family’s interaction and provides the framework for research on the molecular functions of CmWOX genes.
Primary versus Secondary Elevations in Fundus Autofluorescence
Authors: Parmann Rait , Tsang Stephen H. , Sparrow Janet R.
Published: August 2023

The method of quantitative fundus autofluorescence (qAF) can be used to assess the levels of bisretinoids in retinal pigment epithelium (RPE) cells so as to aid the interpretation and management of a variety of retinal conditions. In this review, we focused on seven retinal diseases to highlight the possible pathways to increased fundus autofluorescence. ABCA4- and RDH12-associated diseases benefit from known mechanisms whereby gene malfunctioning leads to elevated bisretinoid levels in RPE cells. On the other hand, peripherin2/RDS-associated disease (PRPH2/RDS), retinitis pigmentosa (RP), central serous chorioretinopathy (CSC), acute zonal occult outer retinopathy (AZOOR), and ceramide kinase like (CERKL)-associated retinal degeneration all express abnormally high fundus autofluorescence levels without a demonstrated pathophysiological pathway for bisretinoid elevation. We suggest that, while a known link from gene mutation to increased production of bisretinoids (as in ABCA4- and RDH12-associated diseases) causes primary elevation in fundus autofluorescence, a secondary autofluorescence elevation also exists, where an impairment and degeneration of photoreceptor cells by various causes leads to an increase in bisretinoid levels in RPE cells.
Multi-Target Neural Differentiation (MTND) Therapeutic Cocktail to Suppress Brain Tumor
Authors: Hu Xiaoping , Xie Jingdun , Yang Yilin , Qiu Ziyi , Lu Weicheng , Lin Xudong , Xu Bingzhe
Published: August 2023

Brain tumors have been proved challenging to treat. Here we established a Multi-Target Neural Differentiation (MTND) therapeutic cocktail to achieve effective and safe treatment of brain malignancies by targeting the important hallmarks in brain cancers: poor cell differentiation and compromised cell cycle. In-vitro and in-vivo experiments confirmed the significant therapeutic effect of our MTND therapy. Significantly improved therapeutic effects over current first-line chemo-drugs have been identified in clinical cells, with great inhibition of the growth and migration of tumor cells. Further in-vivo experiments confirmed that sustained MTND treatment showed a 73% reduction of the tumor area. MTND also induced strong expression of phenotypes associated with cell cycle exit/arrest and rapid neural reprograming from clinical glioma cells to glutamatergic and GABAergic expressing cells, which are two key neuronal types involved in many human brain functions, including learning and memory. Collectively, MTND induced multi-targeted genotypic expression changes to achieve direct neural conversion of glioma cells and controlled the cell cycle/tumorigenesis development, helping control tumor cells’ malignant proliferation and making it possible to treat brain malignant tumors effectively and safely. These encouraging results open avenues to developing new therapies for brain malignancies beyond cytotoxic agents, providing more effective medication recommendations with reduced toxicity.
Improved Prostate-Specific Membrane Antigen (PSMA) Stimulation Using a Super Additive Effect of Dutasteride and Lovastatin In Vitro
Authors: Kuzmanov Aleksandar , Salemi Souzan , Schmid Florian A. , Burger Irene A. , Eberli Daniel , Kranzbühler Benedikt
Published: August 2023

Prostate-specific membrane antigen (PSMA)-based imaging improved the detection of primary, recurrent and metastatic prostate cancer. However, in certain patients, a low PSMA surface expression can be a limitation for this promising diagnostic tool. Pharmacological induction of PSMA might be useful to further improve the detection rate of PSMA-based imaging. To achieve this, we tested dutasteride (Duta)—generally used for treatment of benign prostatic enlargement—and lovastatin (Lova)—a compound used to reduce blood lipid concentrations. We aimed to compare the individual effects of Duta and Lova on cell proliferation as well as PSMA expression. In addition, we tested if a combination treatment using lower concentrations of Duta and Lova can further induce PSMA expression. Our results show that a treatment with ≤1 μM Duta and ≥1 μM Lova lead to a significant upregulation of whole and cell surface PSMA expression in LNCaP, C4-2 and VCaP cells. Lower concentrations of Duta and Lova in combination (0.5 μM Duta + 0.5 μM Lova or 0.5 μM Duta + 1 μM Lova) were further capable of enhancing PSMA protein expression compared to a single compound treatment using higher concentrations in all tested cell lines (LNCaP, C4-2 and VCaP).
Short Culture of Bovine Hepatocytes Biopsied from Dairy Cows as a Model for Toxicological Studies—CYP 1A1 Activity Response to Zearalenone Treatment
Authors: Barć Justyna , Flaga Jadwiga , Kozubek Anna , Kowalski Zygmunt Maciej
Published: August 2023

This study presents a simple and cost-effective method for isolating hepatocytes from liver biopsies obtained from healthy and ketotic dairy cows, which can be utilized for studying cellular metabolism, drug toxicity, and hepatocyte-specific gene function and regulation. The expression of hepatocyte marker genes (G6PC, ALB, CYP1A2) was measured and found to be highest at 6 h post-isolation, with a subsequent decrease over time. Cells isolated from ketotic livers exhibited lower expression levels than those from healthy livers. Furthermore, for the functional characterization of ketotic hepatocytes, the cells were exposed to varying doses of zearalenone (ZEA). While doses of 10–50 µM did not affect cell viability, the highest dose of ZEA (100 µM) significantly decreased cell viability, as measured using XTT assay. Additionally, the potential induction of cytochrome P450 A1 (CYP1A1) by ZEA was found. Despite limitations such as a short-term culture, this model provides a useful tool for conducting toxicological research.
The Roles of Hormone Signals Involved in Rhizosphere Pressure Response Induce Corm Expansion in Sagittaria trifolia
Authors: Li Enjiao , Tang Jing , Liu Jiexia , Zhang Zhiping , Hua Bing , Jiang Jiezeng , Miao Minmin
Published: August 2023

Soil is the base for conventional plant growth. The rhizosphere pressure generated from soil compaction shows a dual effect on plant growth in agricultural production. Compacted soil leads to root growth stagnation and causes bending or thickening, thus affecting the growth of aboveground parts of plants. In arrowhead (Sagittaria trifolia L.), the corms derived from the expanded tips of underground stolons are its storage organ. We found that the formation of corms was significantly delayed under hydroponic conditions without rhizosphere pressure originating from soil/sand. In the initial stage of corm expansion, the anatomic structure of arrowhead corm-forming parts harvested from hydroponics and sand culture was observed, and we found that the corm expansion was derived from cell enlargement and starch accumulation. Comparative transcriptome analysis indicated that the corm expansion was closely related to the change in endogenous hormone levels. Endogenous abscisic acid and salicylic acid concentrations were significantly increased in sand-cultured corms. Higher ethylene and jasmonic acid contents were also detected in all arrowhead samples, demonstrating that these hormones may play potential roles in the rhizosphere pressure response and corm expansion. The expression of genes participating in hormone signaling could explain the rising accumulation of certain hormones. Our current results draw an extensive model to reveal the potential regulation mechanism of arrowhead corm expansion promoted by rhizosphere pressure, which will provide important references for further studying the molecular mechanism of rhizosphere pressure modulating the development of underground storage organs in other plants.
Humanized NSG Mouse Models as a Preclinical Tool for Translational Research in Inflammatory Bowel Diseases
Authors: Weß Veronika , Schuster-Winkelmann Paula , Karatekin Yasemin Hazal , Malik Simge , Beigel Florian , Kühn Florian , Gropp Roswitha
Published: August 2023

The development of animal models reflecting the pathologies of ulcerative colitis (UC) and Crohn’s disease (CD) remains a major challenge. The NOD/SCID/IL2rγnull (NSG) mouse strain, which is immune-compromised, tolerates the engraftment of human peripheral blood mononuclear cells (PBMC) derived from patients with UC (NSG-UC) or CD (NSG-CD). This offers the opportunity to examine the impact of individual immunological background on the development of pathophysiological manifestations. When challenged with ethanol, NSG-UC mice exhibited a strong pro-inflammatory response, including the development of edemas, influx of human T cells, B cells and monocytes into the mucosa and submucosa, and elevated expression of the inflammatory markers CRP and CCL-7. Fibrotic alterations were characterized by an influx of fibroblasts and a thickening of the muscularis mucosae. In contrast, the development of pathological manifestations in NSG-CD mice developed without challenge and was signified by extensive collagen deposition between the muscularis propria and muscularis mucosae, as observed in the areas of strictures in CD patients. Vimentin-expressing fibroblasts supplanting colonic crypts and elevated expression of HGF and TGFß corroborated the remodeling phenotype. In summary, the NSG-UC and NSG-CD models partially reflect these human diseases and are powerful tools to examine the mechanism underlying the inflammatory processes in UC and CD.
Stages of NETosis Development upon Stimulation of Neutrophils with Activators of Different Types
Authors: Inozemtsev Vladimir , Sergunova Viktoria , Vorobjeva Nina , Kozlova Elena , Sherstyukova Ekaterina , Lyapunova Snezhanna , Chernysh Aleksandr
Published: August 2023

Before NETs are released, the neutrophil undergoes structural changes. First, it flattens, accompanied by a change in cell shape and rearrangement of the cytoskeleton. Then, nuclear swelling begins, which ends with the ejection of NETs into the extracellular space. We used widefield and confocal fluorescence microscopy to register morphological and structural changes in neutrophils during activation and NETosis. Different types of activators were used, such as NOX-dependent PMA and calcium ionophore A23187. The measurements were performed in a series of sequential stages. In the first stage (30 s after addition of activators and immediately after stimulation of neutrophils), the response of neutrophils to A23187 and PMA exposure was studied. Subsequently, the characteristics of neutrophils in different phases of activation were examined over a longer period of time (30, 60, 120, 180, and 240 min). The specific features of NETosis development were analyzed separately. During the first 30 s, neutrophils appeared to be heterogeneous in shape and structure of the actin cytoskeleton. Characteristic cell shapes included 30″ type 1 cells, similar in shape to the control, with F-actin concentrated in the center of the cytoplasm, and 30″ type 2 cells, which had flattened (spread) shapes with increased frontal dimensions and F-actin distributed throughout the cell. Later, the development of nuclear swelling, the corresponding changes in neutrophil membranes, and NET release into the extracellular space were evaluated. The conditions determining the initiation of chromatin ejection and two characteristic types of decondensed chromatin ejection were revealed. The results obtained contribute to a better understanding of the biophysical mechanisms of neutrophil activation and NETosis development.
Protein Modelling Highlighted Key Catalytic Sites Involved in Position-Specific Glycosylation of Isoflavonoids
Authors: Sajid Moon , Kaur Parwinder
Published: August 2023

Uridine diphosphate glycosyltransferases (UGTs) are known for promiscuity towards sugar acceptors, a valuable characteristic for host plants but not desirable for heterologous biosynthesis. UGTs characterized for the O-glycosylation of isoflavonoids have shown a variable efficiency, substrate preference, and OH site specificity. Thus, 22 UGTs with reported isoflavonoid O-glycosylation activity were analyzed and ranked for OH site specificity and catalysis efficiency. Multiple-sequence alignment (MSA) showed a 33.2% pairwise identity and 4.5% identical sites among selected UGTs. MSA and phylogenetic analysis highlighted a comparatively higher amino acid substitution rate in the N-terminal domain that likely led to a higher specificity for isoflavonoids. Based on the docking score, OH site specificity, and physical and chemical features of active sites, selected UGTs were divided into three groups. A significantly high pairwise identity (67.4%) and identical sites (31.7%) were seen for group 1 UGTs. The structural and chemical composition of active sites highlighted key amino acids that likely define substrate preference, OH site specificity, and glycosylation efficiency towards selected (iso)flavonoids. In conclusion, physical and chemical parameters of active sites likely control the position-specific glycosylation of isoflavonoids. The present study will help the heterologous biosynthesis of glycosylated isoflavonoids and protein engineering efforts to improve the substrate and site specificity of UGTs.
Novel Mutation Glu98Lys in Cardiac Tropomyosin Alters Its Structure and Impairs Myocardial Relaxation
Authors: Matyushenko Alexander M. , Nefedova Victoria V. , Kochurova Anastasia M. , Kopylova Galina V. , Koubassova Natalia A. , Shestak Anna G. , Yampolskaya Daria S. , Shchepkin Daniil V. , Kleymenov Sergey Y. , Ryabkova Natalia S. , Katrukha Ivan A. , Bershitsky Sergey Y. , Zaklyazminskaya Elena V. , Tsaturyan Andrey K. , Levitsky Dmitrii I.
Published: August 2023

We characterized a novel genetic variant c.292G > A (p.E98K) in the TPM1 gene encoding cardiac tropomyosin 1.1 isoform (Tpm1.1), found in a proband with a phenotype of complex cardiomyopathy with conduction dysfunction and slow progressive neuromuscular involvement. To understand the molecular mechanism by which this mutation impairs cardiac function, we produced recombinant Tpm1.1 carrying an E98K substitution and studied how this substitution affects the structure of the Tpm1.1 molecule and its functional properties. The results showed that the E98K substitution in the N-terminal part of the Tpm molecule significantly destabilizes the C-terminal part of Tpm, thus indicating a long-distance destabilizing effect of the substitution on the Tpm coiled-coil structure. The E98K substitution did not noticeably affect Tpm’s affinity for F-actin but significantly impaired Tpm’s regulatory properties. It increased the Ca2+ sensitivity of the sliding velocity of regulated thin filaments over cardiac myosin in an in vitro motility assay and caused an incomplete block of the thin filament sliding at low Ca2+ concentrations. The incomplete motility block in the absence of Ca2+ can be explained by the loosening of the Tpm interaction with troponin I (TnI), thus increasing Tpm mobility on the surface of an actin filament that partially unlocks the myosin binding sites. This hypothesis is supported by the molecular dynamics (MD) simulation that showed that the E98 Tpm residue is involved in hydrogen bonding with the C-terminal part of TnI. Thus, the results allowed us to explain the mechanism by which the E98K Tpm mutation impairs sarcomeric function and myocardial relaxation.
Oxidative Stress Response Kinetics after 60 Minutes at Different (1.4 ATA and 2.5 ATA) Hyperbaric Hyperoxia Exposures
Authors: Leveque Clément , Mrakic Sposta Simona , Theunissen Sigrid , Germonpré Peter , Lambrechts Kate , Vezzoli Alessandra , Bosco Gerardo , Lévénez Morgan , Lafère Pierre , Guerrero François , Balestra Costantino
Published: August 2023

Hyperbaric oxygen therapy (HBOT) is a therapeutical approach based on exposure to pure oxygen in an augmented atmospheric pressure. Although it has been used for years, the exact kinetics of the reactive oxygen species (ROS) between different pressures of hyperbaric oxygen exposure are still not clearly evidenced. In this study, the metabolic responses of hyperbaric hyperoxia exposures for 1 h at 1.4 and 2.5 ATA were investigated. Fourteen healthy non-smoking subjects (2 females and 12 males, age: 37.3 ± 12.7 years old (mean ± SD), height: 176.3 ± 9.9 cm, and weight: 75.8 ± 17.7 kg) volunteered for this study. Blood samples were taken before and at 30 min, 2 h, 24 h, and 48 h after a 1 h hyperbaric hyperoxic exposure. The level of oxidation was evaluated by the rate of ROS production, nitric oxide metabolites (NOx), and the levels of isoprostane. Antioxidant reactions were assessed through measuring superoxide dismutase (SOD), catalase (CAT), cysteinylglycine, and glutathione (GSH). The inflammatory response was measured using interleukine-6, neopterin, and creatinine. A short (60 min) period of mild (1.4 ATA) and high (2.5 ATA) hyperbaric hyperoxia leads to a similar significant increase in the production of ROS and antioxidant reactions. Immunomodulation and inflammatory responses, on the contrary, respond proportionally to the hyperbaric oxygen dose. Further research is warranted on the dose and the inter-dose recovery time to optimize the potential therapeutic benefits of this promising intervention.
Personalized CFTR Modulator Therapy for G85E and N1303K Homozygous Patients with Cystic Fibrosis
Authors: Graeber Simon Y. , Balázs Anita , Ziegahn Niklas , Rubil Tihomir , Vitzthum Constanze , Piehler Linus , Drescher Marika , Seidel Kathrin , Rohrbach Alexander , Röhmel Jobst , Thee Stephanie , Duerr Julia , Mall Marcus A. , Stahl Mirjam
Published: August 2023

CFTR modulator therapy with elexacaftor/tezacaftor/ivacaftor (ETI) has been approved for people with CF and at least one F508del allele in Europe. In the US, the ETI label has been expanded to 177 rare CFTR mutations responsive in Fischer rat thyroid cells, including G85E, but not N1303K. However, knowledge on the effect of ETI on G85E or N1303K CFTR function remains limited. In vitro effects of ETI were measured in primary human nasal epithelial cultures (pHNECs) of a G85E homozygous patient and an N1303K homozygous patient. Effects of ETI therapy in vivo in these patients were assessed using clinical outcomes, including multiple breath washout and lung MRI, and the CFTR biomarkers sweat chloride concentration (SCC), nasal potential difference (NPD) and intestinal current measurement (ICM), before and after initiation of ETI. ETI increased CFTR-mediated chloride transport in G85E/G85E and N1303K/N1303K pHNECs. In the G85E/G85E and the N1303K/N1303K patient, we observed an improvement in lung function, SCC, and CFTR function in the respiratory and rectal epithelium after initiation of ETI. The approach of combining preclinical in vitro testing with subsequent in vivo verification can facilitate access to CFTR modulator therapy and enhance precision medicine for patients carrying rare CFTR mutations.
Sustainable Solution for Plastic Pollution: Upcycling Waste Polypropylene Masks for Effective Oil-Spill Management
Authors: Saleem Junaid , Moghal Zubair Khalid Baig , Shakoor Rana Abdul , McKay Gordon
Published: August 2023

The use of Polypropylene PP in disposable items such as face masks, gloves, and personal protective equipment has increased exponentially during and after the COVID-19 pandemic, contributing significantly to microplastics and nanoplastics in the environment. Upcycling of waste PP provides a useful alternative to traditional thermal and mechanical recycling techniques. It transforms waste PP into useful products, minimizing its impact on the environment. Herein, we synthesized an oil-sorbent pouch using waste PP, which comprises superposed microporous and fibrous thin films of PP using spin coating. The pouch exhibited super-fast uptake kinetics and reached its saturation in fewer than five minutes with a high oil uptake value of 85 g/g. Moreover, it displayed high reusability and was found to be effective in absorbing oil up to seven times when mechanically squeezed between each cycle, demonstrating robust oil-sorption capabilities. This approach offers a potential solution for managing plastic waste while promoting a circular economy.
Tetracyclic 1,4-Naphthoquinone Thioglucoside Conjugate U-556 Blocks the Purinergic P2X7 Receptor in Macrophages and Exhibits Anti-Inflammatory Activity In Vivo
Authors: Kozlovskiy Sergei , Pislyagin Evgeny , Menchinskaya Ekaterina , Chingizova Ekaterina , Kaluzhskiy Leonid , Ivanov Alexis S. , Likhatskaya Galina , Agafonova Irina , Sabutski Yuri , Polonik Sergey , Manzhulo Igor , Aminin Dmitry
Published: August 2023

P2X7 receptors (P2X7Rs) are ligand-gated ion channels that play a significant role in inflammation and are considered a potential therapeutic target for some inflammatory diseases. We have previously shown that a number of synthetic 1,4-naphthoquinones are capable of blocking P2X7Rs in neuronal and macrophage cells. In the present investigation, we have demonstrated the ability of the tetracyclic quinone-thioglucoside conjugate U-556, derived from 1,4-naphthoquinone thioglucoside, to inhibit ATP-induced Ca2+ influx and YO-PRO-1 dye uptake, which indicates blocking P2X7R in RAW 264.7 macrophages. This process was accompanied by the inhibition of ATP-induced reactive oxygen species production in macrophages, as well as the macrophage survival strengthening under ATP toxic effects. Nevertheless, U-556 had no noticeable antioxidant capacity. Naphthoquinone-thioglucoside conjugate U-556 binding to the extracellular part of the P2X7R was confirmed by SPR analysis, and the kinetic characteristics of this complex formation were established. Computer modeling predicted that U-556 binds the P2X7R allosteric binding site, topographically similar to that of the specific A438079 blocker. The study of biological activity in in vivo experiments shows that tetracylic conjugate significantly reduces inflammation provoked by carrageenan. The data obtained points out that the observed physiological effects of U-556 may be due to its ability to block the functioning of the P2X7R.
Shadow of a Pandemic: Persistence of Prenatal SARS-CoV-2 Antibodies in Newborn Blood Spots
Authors: Sciortino Stanley , Graham Steve , Fillman Toki , Kandasamy Hari , Cooley Robin , Hanson Carl , Eckert Valorie , Tang Hao , Yang Juan , Seftel David , Tsai Cheng-ting , Robinson Peter
Published: August 2023

To investigate COVID-19 surveillance among pregnant women, the California Genetic Disease Screening Program conducted a screening performance and seroprevalence evaluation of maternal SARS-CoV-2 antibodies detected in banked newborn dried blood spots (DBS). We obtained seropositive results for 2890 newborn DBS from cohorts in 2020 and 2021 using Enable Bioscience’s Antibody Detection by Agglutination-PCR (ADAP) assay for SARS-CoV-2 antibodies. To infer maternal infection, we linked 312 women with a known laboratory-confirmed COVID-19 episode with their newborn’s DBS SARS-CoV-2 antibody result. Among 2890 newborns, we detected 453 (15.7%) with SARS-CoV-2 antibodies in their DBS. Monthly snapshot statewide seroprevalence among neonates was 12.2% (95% CI 10.3–14.1%, n =1156) in December 2020 and 33.3% (95% CI 29.1–37.4%, n = 26) in March 2021. The longest time recorded from COVID-19 infection to a seropositive neonatal result was 11.7 months among the 312 mothers who had an available SARS-CoV-2 PCR test result. Approximately 94% (153/163) of DBS were seropositive when a known maternal infection occurred earlier than 19 days before birth. The estimated relative sensitivity of DBS to identify prevalent maternal infection was 85.1%, specificity 98.5% and PPV 99.2% (n = 312); the sensitivity was lowest during the December 2021 surge when many infections occurred within 19 days of birth. Fifty pre-pandemic specimens (100% seronegative) and 23 twin-pair results (100% concordant) support an intrinsic specificity and PPV of ADAP approaching 100%. Maternal infection surveillance is limited by a time lag prior to delivery, especially during pandemic surges.