bluuebunny/crossref_metadata_2025 huggingface dataset:
Water Stress-Induced Divergence Growth of Picea schrenkiana in the Western Tianshan and Its Forcing Mechanisms
Authors: Li Teng , Liu Yu , Cai Qiufang , Duan Xiangyu , Li Pei , Ren Meng , Ye Yuanda
Published: February 2023

Since the 1950s, divergence problems have reduced the temporal stability of tree rings in response to climate, shaken the foundations of dendroclimatology, and affected the reliability of reconstructed models based on tree rings and the accuracy of historical climate series. Therefore, it is of great importance to investigate divergence problems, which will help us to better understand the growth strategies of trees in response to climate warming and provide a scientific basis for accurate climate reconstruction and simulation of forest dynamics. This paper aims to elucidate the mechanism of divergent growth of Picea schrenkiana at high altitudes in the western Tianshan from three aspects: variations in atmospheric circulation, changes in climatic factors, and the coping strategy of trees with climate change. High spring temperatures accelerate the melting of snow cover. Large amounts of snowmelt initially replenish soil water, leading to the rapid growth of trees, but later, the continuous consumption of snowmelt reduces the available water capacity of soil, resulting in water stress on trees. The pattern of trees’ response to changes in climate ranges from a pure temperature limitation to a collaborative temperature–moisture limitation. Since the 1990s, the weakening of the westerly circulation and the North Atlantic Oscillation has reduced their impacts on the hydroclimate in the Tianshan Mountains of Central Asia. The combination of heat-induced water deficit and a long-term weakening trend in atmospheric circulation has slowed tree growth over the past decade.
Designing for the Metaverse: A Multidisciplinary Laboratory in the Industrial Design Program
Authors: Ricci Marina , Scarcelli Alessandra , Fiorentino Michele
Published: February 2023

The design research and education landscapes are changing. The widespread development and use of technologies such as Mixed Reality (MR) and the diffusion of Head-Mounted Displays (HMDs) available at low cost are causing a shift in design education toward the Metaverse. In this ever-changing scenario, there is a need to rethink design and teaching methods. However, scientific literature lacks the ability to provide contributions that include MR technology education in the industrial design program. We, therefore, present an innovative laboratory with an integrated multidisciplinary approach that starts from the fundamentals of interaction design and aims to teach students how to design next-generation MR interfaces for the Metaverse. The lab combines theory and practice within three courses: Information Design, Information Systems, and Virtual Design and Simulation. Industrial design students follow a precise multidisciplinary method consisting of five steps, from state-of-the-art analysis to the presentation of a final group design of an MR user interface. Thus, we introduce a class case study by presenting the outcomes of a semester project in the field of household appliances. Evaluation of the teaching method is conducted through a semi-structured questionnaire. Preliminary results show positive outcomes from students in terms of acceptance, effectiveness, usefulness, efficiency, and satisfaction with the teaching method adopted for the laboratory.
Comparative Hypothalamic Transcriptome Analysis Reveals Crucial mRNAs, lncRNAs, and circRNAs Affecting Litter Size in Goats
Authors: Dong Shucan , Hou Biwei , Yang Chuang , Li Yaokun , Sun Baoli , Guo Yongqing , Deng Ming , Liu Dewu , Liu Guangbin
Published: February 2023

Litter size is an important indicator to measure the reproductive performance of goats, which is affected by the reproductive function of animals. The hypothalamus, as the regulatory center of the endocrine system, plays an important role in the reproduction of female animals. Here, we performed high-throughput RNA sequencing using hypothalamic tissue from high-fecundity and low-fecundity Leizhou goats to explore critical functional genes associated with litter size. Differentially expressed mRNA, lncRNA, and circRNAs were screened using DESeq and were enriched, and then analyzed by Gene Ontology and Kyoto Encyclopedia of Gene and Genome. Results showed that some of these differentially expressed mRNAs could be enriched in reproductive processes, jak-STAT, prolactin signaling pathway, and other signaling pathways related to reproduction, such as SOCS3. Furthermore, the central proteins POSTN, MFAP5, and DCN from protein–protein interaction may regulate animal reproductive activity by affecting cell proliferation and apoptosis. lncRNA MSTRG.33887.2 as well as circRNAs chicirc_098002, chicirc_072583, and chicirc_053531 may be able to influence animal reproduction by participating in folate metabolism and energy metabolism homeostasis through their respective target genes. Our results expand the molecular mechanism of hypothalamic regulation on animal reproduction.
Effects of Elexacaftor/Tezacaftor/Ivacaftor on Cardiorespiratory Polygraphy Parameters and Respiratory Muscle Strength in Cystic Fibrosis Patients with Severe Lung Disease
Authors: Giallongo Alessandro , Parisi Giuseppe Fabio , Papale Maria , Manti Sara , Mulé Enza , Aloisio Donatella , Terlizzi Vito , Rotolo Novella , Leonardi Salvatore
Published: February 2023

Background: Cystic fibrosis transmembrane conductance regulator (CFTR) modulators represent targeted therapies directly acting on the CFTR channel. The triple therapy Elexacaftor/Tezacaftor/Ivacaftor (ELX/TEZ/IVA) has been demonstrated to improve lung function and quality of life in cystic fibrosis (CF) patients. However, the effects of ELX/TEZ/IVA on sleep-disordered breathing (SDB) and respiratory muscle strength are poorly studied. The aim of this study was to assess the effects of ELX/TEZ/IVA in patients with CF and severe lung disease on cardiorespiratory polygraphy parameters, maximum inspiratory pressure (MIP) and maximum expiratory pressure (MEP) measures. Methods: patients with CF aged ≥ 12 who started treatment in a compassionate use program were retrospectively studied through the evaluation of nocturnal cardiorespiratory polygraphy parameters, MIP and MEP; and six-minute walk test (6MWT) at baseline and at months 3, 6, and 12 of treatment. Results: Nine patients (mean age 30.3 ± 6.5 years) with severe CF (mean baseline ppFEV1 34.6 ± 5.1%) were evaluated. A significant improvement in nocturnal oxygenation measured by mean SpO2 (92.4 vs. 96.4%, p < 0.05), time spent with SpO2 ≤ 90% (−12.6, −14.6, −15.2 min from baseline at months 3, 6, and 12, respectively, p < 0.05), and respiratory rate (RR) was shown, at month 12 and across the time points compared with baseline, as well as in respiratory muscle strength, although only the change in MEP was significant. Conclusions: We provide further evidence on the efficacy of the CFTR modulators ELX/TEZ/IVA, adding information about their effect on the respiratory muscles’ performance and cardiorespiratory polygraphy parameters in CF patients with severe lung disease.
Impact of pri-let-7a-1 rs10739971 for Gastric Cancer Predisposition in an Amazon Region
Authors: Andrade Roberta Borges , Cohen-Paes Amanda de Nazaré , Leal Diana Feio da Veiga Borges , Pantoja Karla Beatriz Cardias Cereja , Gellen Laura Patrícia Albarello , Carvalho Darlen Cardoso de , Piedade de Souza Tatiane , Fernandes Marianne Rodrigues , Assumpcão Paulo Pimentel de , Burbano Rommel Mario Rodríguez , Santos Sidney Emanuel Batista dos , Santos Ney Pereira Carneiro dos
Published: February 2023

Gastric cancer (GC) is the fifth most common type of cancer and the fourth leading cause of cancer death. In Brazil, GC has a high incidence and mortality rates, and it is highly variable by region. The Amazon region has significant rising rates among all Brazil regions. Only very few studies have evaluated the association between genetic variants and the risk of gastric cancer in the Brazilian Amazon population. Therefore, this study aimed to investigate associations between single nucleotide polymorphisms of miRNA processing genes and the risk for GC in this population. Potentially functional single nucleotide polymorphisms from miRNA processing genes were genotyped in 159 cases and 193 healthy controls by QuantStudio Real Time PCR. According to our findings, the genotype GG of the variant rs10739971 presents a lower risk to the development of GC in comparison to the remaining genotypes (p = 0.000016; OR = 0.055; 95% CI = 0.015–0.206). This is the first study to report the association of pri-let-7a-1 rs10739971 with GC in the Brazilian Amazon population, which is a highly mixed population with a unique genetic constitution that is different from other populations that are studied in the vast majority of scientific research.
Prediction of Work from Home and Musculoskeletal Discomfort: An Investigation of Ergonomic Factors in Work Arrangements and Home Workstation Setups Using the COVID-19 Experience
Authors: Chim Justine M. Y. , Chen Tien Li
Published: February 2023

The COVID-19 pandemic provided an opportunity for office workers to experience work from home (WFH). The aims of this study are to investigate the prevalence rate of musculoskeletal discomfort (MSD) and the work conditions of homeworkers during WFH as well as to evaluate the association and predicted risk of ergonomic factors and MSD. A total of 232 homeworkers completed questionnaires. Chi-square test and logistic regression were used to analyze the association and prediction of work arrangements and home workstation setups and musculoskeletal outcomes. The result showed that 61.2% of homeworkers reported MSD while WFH. Because of the small living spaces in Hong Kong, 51% and 24.6% of homeworkers worked in living/dining areas and bedrooms, respectively, potentially affecting their work and personal life. Additionally, homeworkers adopted a flexible work style, but prolonged computer use while WFH. Homeworkers who used a chair without a backrest or a sofa could predict a significantly higher risk of MSD. The use of a laptop monitor posed about a 2 to 3 times higher risk of suffering from neck, upper back, and lower back discomfort than the use of a desktop monitor. These results provide valuable information to help regulators, employers, homeworkers, and designers create better WFH guidelines, work arrangements, and home settings.
Efficiency Recycling and Utilization of Phosphate from Wastewater Using LDHs-Modified Biochar
Authors: Ding Chunxia , Long Xiuyu , Zeng Guangyong , Ouyang Yu , Lei Bowen , Zeng Rongying , Wang Jing , Zhou Zhi
Published: February 2023

The excessive application of phosphate fertilizers easily causes water eutrophication. Phosphorus recovery by adsorption is regarded as an effective and simple intervention to control water bodies’ eutrophication. In this work, a series of new adsorbents, layered double hydroxides (LDHs)-modified biochar (BC) with different molar ratios of Mg2+ and Fe3+, were synthesized based on waste jute stalk and used for recycling phosphate from wastewater. The prepared LDHs-BC4 (the molar ratio of Mg/Fe is 4:1) has significantly high adsorption performance, and the recovery rate of phosphate is about 10 times higher than that of the pristine jute stalk BC. The maximum adsorption capacity of LDHs-BC4 for phosphate was 10.64 mg-P/g. The main mechanism of phosphate adsorption mainly includes electrostatic attraction, ion exchange, ligand exchange, and intragranular diffusion. Moreover, the phosphate-adsorbed LDHs-BC4 could promote mung bean growth, which indicated the recovery phosphate from wastewater could be used as a fertilizer.
Facile Phase Control and Photocatalytic Performance of BiVO4 Crystals for Methylene Blue Degradation
Authors: Cai Heshan , Cheng Linmei , Chen Huacong , Dou Rongni , Chen Junfeng , Zhao Yuxin , Li Fuhua , Fang Zheng
Published: February 2023

Emerging contaminants, which mainly exist as organic pollutants and pose adverse biological effects, could be removed using photocatalytic degradation, resulting in a low-cost and environmentally friendly solution. Herein, BiVO4 nanoparticles with different morphologies and photocatalytic performances were synthesized by hydrothermal treatment at different residence times. The XRD and SEM results indicate that the crystal phase of BiVO4 gradually transformed from a single tetragonal phase to a single monoclinic crystal phase as the hydrothermal time increased, and with the extension of the hydrothermal time, the morphology of BiVO4 nanoparticles gradually differentiated from a smooth spherical shape to flower-like shapes composed of polyhedrons; the size of the crystals also increased accordingly. Methylene blue (MB), used as a probe of organic pollutants, was degraded under visible light irradiation by all BiVO4 samples to investigate its photocatalytic activities. The experimental results show that the longer the hydrothermal time, the better the photocatalytic performance. The optimum hydrothermal time was 24 h, at which the sample showed the highest photocatalytic activity for MB degradation. This work shows a convenient strategy for control of the crystal phase of BiVO4-based photocatalysts based on the understanding of the crystal morphology evolution mechanism, which will benefit the researchers in designing new BiVO4-based photocatalysts with high efficiency for emerging contaminants’ degradation.
A Comparative Analysis of E-Cigarette and Cigarette Posts on Instagram
Authors: Lim Young-shin , Lee Ji Young
Published: February 2023

E-cigarette use has grown rapidly over the past decade and become a threat to public health. Marketing—especially through social media—has contributed significantly to this growth, which suggests that regulating content in social media will be critical in supporting efforts to reverse this trend. A content analysis was performed to compare 254 e-cigarette posts on Instagram with 228 cigarette posts on the same platform. The majority of e-cigarette posts were from e-cigarette companies (40.9%) and industry people (18.5%), whereas the majority of cigarette posts were from laypeople (76.8%). More e-cigarette posts than cigarette posts appeared to have a marketing intent (56.3% vs. 1.3%), and brand representation in photographs/videos was more frequent in the e-cigarette posts than in the cigarette posts (63.0% vs. 15.8%). Further, compared with the e-cigarette posts, the cigarette posts were more likely to portray daily life (73.2% vs. 41.3%) and humans (80.3% vs. 43.7%) in the photograph/video. The cigarette posts also portrayed smoking much more often than the e-cigarette posts portrayed vaping (67.1% vs. 21.3%). The study findings broaden the field’s understanding of cigarette and e-cigarette content on Instagram and social media, and have implications for monitoring and regulating content for e-cigarettes and cigarettes.
Extracellular Vesicle Treatment Alleviates Neurodevelopmental and Neurodegenerative Pathology in Cortical Spheroid Model of Down Syndrome
Authors: Campbell Natalie Baker , Patel Yesha , Moore Tara L. , Medalla Maria , Zeldich Ella
Published: February 2023

Down syndrome (DS), or trisomy 21, is manifested in a variety of anatomical and cellular abnormalities resulting in intellectual deficits and early onset of Alzheimer’s disease (AD) with no effective treatments available to alleviate the pathologies associated with the disorder. The therapeutic potential of extracellular vesicles (EVs) has emerged recently in relation to various neurological conditions. We have previously demonstrated the therapeutic efficacy of mesenchymal stromal cell-derived EVs (MSC-EVs) in cellular and functional recovery in a rhesus monkey model of cortical injury. In the current study, we evaluated the therapeutic effect of MSC-EVs in a cortical spheroid (CS) model of DS generated from patient-derived induced pluripotent stem cells (iPSCs). Compared to euploid controls, trisomic CS display smaller size, deficient neurogenesis, and AD-related pathological features, such as enhanced cell death and depositions of amyloid beta (Aβ) and hyperphosphorylated tau (p-tau). EV-treated trisomic CS demonstrated preserved size, partial rescue in the production of neurons, significantly decreased levels of Aβ and p-tau, and a reduction in the extent of cell death as compared to the untreated trisomic CS. Together, these results show the efficacy of EVs in mitigating DS and AD-related cellular phenotypes and pathological depositions in human CS.
Accurate Detection of SARS-CoV-2 by Next-Generation Sequencing in Low Viral Load Specimens
Authors: Ilié Marius , Benzaquen Jonathan , Hofman Véronique , Long-Mira Elodie , Lassalle Sandra , Boutros Jacques , Bontoux Christophe , Lespinet-Fabre Virginie , Bordone Olivier , Tanga Virginie , Allegra Maryline , Salah Myriam , Fayada Julien , Leroy Sylvie , Vassallo Matteo , Touitou Irit , Courjon Johan , Contenti Julie , Carles Michel , Marquette Charles-Hugo , Hofman Paul
Published: February 2023

As new SARS-CoV-2 variants emerge, there is an urgent need to increase the efficiency and availability of viral genome sequencing, notably to detect the lineage in samples with a low viral load. SARS-CoV-2 genome next-generation sequencing (NGS) was performed retrospectively in a single center on 175 positive samples from individuals. An automated workflow used the Ion AmpliSeq SARS-CoV-2 Insight Research Assay on the Genexus Sequencer. All samples were collected in the metropolitan area of the city of Nice (France) over a period of 32 weeks (from 19 July 2021 to 11 February 2022). In total, 76% of cases were identified with a low viral load (Ct ≥ 32, and ≤200 copies/µL). The NGS analysis was successful in 91% of cases, among which 57% of cases harbored the Delta variant, and 34% the Omicron BA.1.1 variant. Only 9% of cases had unreadable sequences. There was no significant difference in the viral load in patients infected with the Omicron variant compared to the Delta variant (Ct values, p = 0.0507; copy number, p = 0.252). We show that the NGS analysis of the SARS-CoV-2 genome provides reliable detection of the Delta and Omicron SARS-CoV-2 variants in low viral load samples.
Positive Linear Relationship between Nucleophosmin Protein Expression and the Viral Load in HPV-Associated Oropharyngeal Squamous Cell Carcinoma: A Possible Tool for Stratification of Patients
Authors: D’Agostino Marco , Di Cecco Marco , Marani Carla , Vigili Maurizio Giovanni , Sileno Sara , Volpi Chiara Costanza , Gloghini Annunziata , Avitabile Daniele , Magenta Alessandra , Rahimi Siavash
Published: February 2023

Most oropharyngeal squamous cell carcinomas (OPSCCs) are human papillomavirus (HPV)-associated, high-risk (HR) cancers that show a better response to chemoradiotherapy and are associated with improved survival. Nucleophosmin (NPM, also called NPM1/B23) is a nucleolar phosphoprotein that plays different roles within the cell, such as ribosomal synthesis, cell cycle regulation, DNA damage repair and centrosome duplication. NPM is also known as an activator of inflammatory pathways. An increase in NPM expression has been observed in vitro in E6/E7 overexpressing cells and is involved in HPV assembly. In this retrospective study, we investigated the relationship between the immunohistochemical (IHC) expression of NPM and HR-HPV viral load, assayed by RNAScope in situ hybridization (ISH), in ten patients with histologically confirmed p16-positive OPSCC. Our findings show that there is a positive correlation between NPM expression and HR-HPV mRNA (Rs = 0.70, p = 0.03), and a linear regression (r2 = 0.55; p = 0.01). These data support the hypothesis that NPM IHC, together with HPV RNAScope, could be used as a predictor of transcriptionally active HPV presence and tumor progression, which is useful for therapy decisions. This study includes a small cohort of patients and, cannot report conclusive findings. Further studies with large series of patients are needed to support our hypothesis.
Exploration of the Core Pathways and Potential Targets of Luteolin Treatment on Late-Onset Depression Based on Cerebrospinal Fluid Proteomics
Authors: Liu Kaige , Li Huizhen , Zeng Ningxi , Li Bozhi , Yao Gaolei , Wu Xiaofeng , Xu Hanfang , Yan Can , Wu Lili
Published: February 2023

Cognitive deficiency is one of the fundamental characteristics of late-onset depression (LOD). Luteolin (LUT) possesses antidepressant, anti-aging, and neuroprotective properties, which can dramatically enhance cognition. The altered composition of cerebrospinal fluid (CSF), which is involved in neuronal plasticity and neurogenesis, directly reflects the physio-pathological status of the central nervous system. It is not well known whether the effect of LUT on LOD is in association with a changed CSF composition. Therefore, this study first established a rat model of LOD and then tested the therapeutic effects of LUT using several behavioral approaches. A gene set enrichment analysis (GSEA) was used to evaluate the CSF proteomics data for KEGG pathway enrichment and Gene Ontology annotation. We combined network pharmacology and differentially expressed proteins to screen for key GSEA–KEGG pathways as well as potential targets for LUT therapy for LOD. Molecular docking was adopted to verify the affinity and binding activity of LUT to these potential targets. The outcomes demonstrated that LUT improved the cognitive and depression-like behaviors in LOD rats. LUT may exert therapeutic effects on LOD through the axon guidance pathway. Five axon guidance molecules—EFNA5, EPHB4, EPHA4, SEMA7A, and NTNG—as well as UNC5B, L1CAM, and DCC, may be candidates for the LUT treatment of LOD.
Anti-Fibrotic Potential of Angiotensin (1-7) in Hemodynamically Overloaded Rat Heart
Authors: Sykora Matus , Kratky Vojtech , Kopkan Libor , Tribulova Narcisa , Szeiffova Bacova Barbara
Published: February 2023

The extracellular matrix (ECM) is a highly dynamic structure controlling the proper functioning of heart muscle. ECM remodeling with enhanced collagen deposition due to hemodynamic overload impairs cardiomyocyte adhesion and electrical coupling that contributes to cardiac mechanical dysfunction and arrhythmias. We aimed to explore ECM and connexin-43 (Cx43) signaling pathways in hemodynamically overloaded rat heart as well as the possible implication of angiotensin (1-7) (Ang (1-7)) to prevent/attenuate adverse myocardial remodeling. Male 8-week-old, normotensive Hannover Spraque–Dawley rats (HSD), hypertensive (mRen-2)27 transgenic rats (TGR) and Ang (1-7) transgenic rats (TGR(A1-7)3292) underwent aortocaval fistula (ACF) to produce volume overload. Five weeks later, biometric and heart tissue analyses were performed. Cardiac hypertrophy in response to volume overload was significantly less pronounced in TGR(A1-7)3292 compared to HSD rats. Moreover, a marker of fibrosis hydroxyproline was increased in both ventricles of volume-overloaded TGR while it was reduced in the Ang (1-7) right heart ventricle. The protein level and activity of MMP-2 were reduced in both ventricles of volume-overloaded TGR/TGR(A1-7)3292 compared to HSD. SMAD2/3 protein levels were decreased in the right ventricle of TGR(A1-7)3292 compared to HSD/TGR in response to volume overload. In parallel, Cx43 and pCx43 implicated in electrical coupling were increased in TGR(A1-7)3292 versus HSD/TGR. It can be concluded that Ang (1-7) exhibits cardio-protective and anti-fibrotic potential in conditions of cardiac volume overload.
Genome-Wide Characterization and Analysis of bHLH Transcription Factors Related to Anthocyanin Biosynthesis in Cinnamomum camphora (‘Gantong 1’)
Authors: Gong Xue , Shen Tengfei , Li Xiuqi , Lin Hanbin , Chen Caihui , Li Huihu , Wu Zhaoxiang , Liu Qiaoli , Xu Meng , Zhang Bo , Zhong Yongda
Published: February 2023

Cinnamomum camphora is one of the most commonly used tree species in landscaping. Improving its ornamental traits, particularly bark and leaf colors, is one of the key breeding goals. The basic helix–loop–helix (bHLH) transcription factors (TFs) are crucial in controlling anthocyanin biosynthesis in many plants. However, their role in C. camphora remains largely unknown. In this study, we identified 150 bHLH TFs (CcbHLHs) using natural mutant C. camphora ‘Gantong 1’, which has unusual bark and leaf colors. Phylogenetic analysis revealed that 150 CcbHLHs were divided into 26 subfamilies which shared similar gene structures and conserved motifs. According to the protein homology analysis, we identified four candidate CcbHLHs that were highly conserved compared to the TT8 protein in A. thaliana. These TFs are potentially involved in anthocyanin biosynthesis in C. camphora. RNA-seq analysis revealed specific expression patterns of CcbHLHs in different tissue types. Furthermore, we verified expression patterns of seven CcbHLHs (CcbHLH001, CcbHLH015, CcbHLH017, CcbHLH022, CcbHLH101, CcbHLH118, and CcbHLH134) in various tissue types at different growth stages using qRT-PCR. This study opens a new avenue for subsequent research on anthocyanin biosynthesis regulated by CcbHLH TFs in C. camphora.
circINSR Inhibits Adipogenic Differentiation of Adipose-Derived Stromal Vascular Fractions through the miR-152/MEOX2 Axis in Sheep
Authors: Zhao Bishi , Zhang Hanyue , Zhao Dan , Liang Yu , Qiao Liying , Liu Jianhua , Pan Yangyang , Yang Kaijie , Liu Wenzhong
Published: February 2023

Adipose tissue plays a crucial role in energy metabolism. Several studies have shown that circular RNA (circRNA) is involved in the regulation of fat development and lipid metabolism. However, little is known about their involvement in the adipogenic differentiation of ovine stromal vascular fractions (SVFs). Here, based on previous sequencing data and bioinformatics analysis, a novel circINSR was identified in sheep, which acts as a sponge to promote miR-152 in inhibiting the adipogenic differentiation of ovine SVFs. The interactions between circINSR and miR-152 were examined using bioinformatics, luciferase assays, and RNA immunoprecipitation. Of note, we found that circINSR was involved in adipogenic differentiation via the miR-152/mesenchyme homeobox 2 (MEOX2) pathway. MEOX2 inhibited adipogenic differentiation of ovine SVFs and miR-152 inhibited the expression of MEOX2. In other words, circINSR directly isolates miR-152 in the cytoplasm and inhibits its ability to promote adipogenic differentiation of ovine SVFs. In summary, this study revealed the role of circINSR in the adipogenic differentiation of ovine SVFs and its regulatory mechanisms, providing a reference for further interpretation of the development of ovine fat and its regulatory mechanisms.
Ejaculatory Abstinence Affects the Sperm Quality in Normozoospermic Men—How Does the Seminal Bacteriome Respond?
Authors: Tvrdá Eva , Ďuračka Michal , Benko Filip , Kováčik Anton , Lovíšek Daniel , Gálová Eliška , Žiarovská Jana , Schwarzová Marianna , Kačániová Miroslava
Published: February 2023

This study was designed to describe bacterial profiles of ejaculates collected following a long and short ejaculatory abstinence set in the context of changes in the conventional, oxidative, and immunological characteristics of semen. Two specimens were collected in succession from normozoospermic men (n = 51) following 2 days and 2 h, respectively. Semen samples were processed and analyzed according to the World Health Organization (WHO) 2021 guidelines. Afterwards, sperm DNA fragmentation, mitochondrial function, levels of reactive oxygen species (ROS), total antioxidant capacity, and oxidative damage to sperm lipids and proteins were evaluated in each specimen. Selected cytokine levels were quantified using the ELISA method. Bacterial identification by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry revealed that samples collected following two days of abstinence presented with a higher bacterial load and diversity, and a greater prevalence of potentially uropathogenic bacteria including Escherichia coli, Staphylococcus aureus and Enterococcus faecalis. Only staphylococci and Escherichia coli remained present in specimens obtained after 2 h of abstinence. Whilst all samples accomplished the criteria set by WHO, a significantly higher motility (p < 0.05), membrane integrity (p < 0.05), mitochondrial membrane potential (p < 0.05), and DNA integrity (p < 0.0001) were detected following 2 h of ejaculatory abstinence. On the other hand, significantly higher ROS levels (p < 0.001), protein oxidation (p < 0.001), and lipid peroxidation (p < 0.01) accompanied by significantly higher concentrations of tumor necrosis factor alpha (p < 0.05), interleukin-6 (p < 0.01), and interferon gamma (p < 0.05) were observed in specimens collected after two days of abstinence. It may be summarized that shorter ejaculatory abstinence does not compromise sperm quality in normozoospermic men, while it contributes to a decreased occurrence of bacteria in semen which is accompanied by a lower probability of damage to spermatozoa by ROS or pro-inflammatory cytokines.
Osteoblastic Cell Behavior and Gene Expression Related to Bone Metabolism on Different Titanium Surfaces
Authors: Velasco-Ortega Eugenio , Fos-Parra Isabel , Cabanillas-Balsera Daniel , Gil Javier , Ortiz-García Iván , Giner Mercè , Bocio-Núñez Jesús , Montoya-García María-José , Jiménez-Guerra Álvaro
Published: February 2023

The surface topography of titanium dental implants has a great influence on osseointegration. In this work, we try to determine the osteoblastic behavior and gene expression of cells with different titanium surfaces and relate them to the physicochemical properties of the surface. For this purpose, we have used commercial titanium discs of grade 3: as-received corresponds to machined titanium without any surface treatment (MA), chemically acid etched (AE), treated via sand blasting with Al2O3 particles (SB) and a sand-blasting treatment with acid etching (SB+AE). The surfaces have been observed using scanning electron microscopy (SEM) and the roughness, wettability and surface energy with dispersive and polar components have been characterized. Osteoblastic cultures were performed with SaOS-2 osteoblastic cells determining cell viability as well as alkaline phosphatase levels for 3 and 21 days, and osteoblastic gene expression was determined. The roughness values of the MA discs was 0.02 μm, which increases to 0.3 μm with acid attack and becomes the maximum for the sand-blasted samples, reaching values of 1.2 μm for SB and SB+AE. The hydrophilic behavior of the MA and AE samples with contact angles of 63° and 65° is superior to that of the rougher samples, being 75° for SB and 82° for SB+AE. In all cases, they show good hydrophilicity. GB and GB+AE surfaces present a higher polar component in the surface energy values, 11.96 and 13.18 mJ/m2, respectively, than AE and MA, 6.64 and 9.79 mJ/m2, respectively. The osteoblastic cell viability values at three days do not show statistically significant differences between the four surfaces. However, the viability of the SB and SB+AE surfaces at 21 days is much higher than that of the AE and MA samples. From the alkaline phosphatase studies, higher values were observed for those treated with sand blasting with and without acid etching compared to the other two surfaces, indicating a greater activity in osteoblastic differentiation. In all cases except in the Osterix (Ostx) —osteoblast-specific transcription factor—a decrease in gene expression is observed in relation to the MA samples (control). The most important increase was observed for the SB+AE condition. A decrease in the gene expression of Osteoprotegerine (OPG), Runt-related transcription factor 2 (Runx2), Receptor Activator of NF-κB Ligand (RANKL) and Alkaline Phosphatase (Alp) genes was observed in the AE surface.
Converging Role for REEP1/SPG31 in Oxidative Stress
Authors: Naef Valentina , Meschini Maria C. , Tessa Alessandra , Morani Federica , Corsinovi Debora , Ogi Asahi , Marchese Maria , Ori Michela , Santorelli Filippo M. , Doccini Stefano
Published: February 2023

Mutations in the receptor expression-enhancing protein 1 gene (REEP1) are associated with hereditary spastic paraplegia type 31 (SPG31), a neurological disorder characterized by length-dependent degeneration of upper motor neuron axons. Mitochondrial dysfunctions have been observed in patients harboring pathogenic variants in REEP1, suggesting a key role of bioenergetics in disease-related manifestations. Nevertheless, the regulation of mitochondrial function in SPG31 remains unclear. To elucidate the pathophysiology underlying REEP1 deficiency, we analyzed in vitro the impact of two different mutations on mitochondrial metabolism. Together with mitochondrial morphology abnormalities, loss-of-REEP1 expression highlighted a reduced ATP production with increased susceptibility to oxidative stress. Furthermore, to translate these findings from in vitro to preclinical models, we knocked down REEP1 in zebrafish. Zebrafish larvae showed a significant defect in motor axon outgrowth leading to motor impairment, mitochondrial dysfunction, and reactive oxygen species accumulation. Protective antioxidant agents such as resveratrol rescued free radical overproduction and ameliorated the SPG31 phenotype both in vitro and in vivo. Together, our findings offer new opportunities to counteract neurodegeneration in SPG31.
Proximate Analysis of Moringa oleifera Leaves and the Antimicrobial Activities of Successive Leaf Ethanolic and Aqueous Extracts Compared with Green Chemically Synthesized Ag-NPs and Crude Aqueous Extract against Some Pathogens
Authors: Ahmed Mostafa , Marrez Diaa Attia , Abdelmoeen Nadia Mohamed , Mahmoud Ebtesam Abdelmoneem , Abdel-Shakur Ali Mohamed , Decsi Kincső , Tóth Zoltán
Published: February 2023

Research on the use of different parts of the Moringa oleifera plant as a nutritional and pharmaceutical resource for human and animals has increased in recent years. This study aimed to investigate the chemical composition and the TPCs and TFCs of Moringa leaves, the antimicrobial activities of Moringa successive ethanolic, aqueous, crude aqueous extracts, and green-chemically synthesized characterized Ag-NPs. The results indicated that the ethanolic extract recorded the highest activity against E. coli. On the other side, the aqueous extract showed higher activity, and its effects ranged from 0.03 to 0.33 mg/mL against different strains. The MIC values of Moringa Ag-NPs against different pathogenic bacteria ranged from 0.05 mg/mL to 0.13 mg/mL, and the activity of the crude aqueous extract ranged from 0.15 to 0.83 mg/mL. For the antifungal activity, the ethanolic extract recorded the highest activity at 0.04 mg/mL, and the lowest activity was recorded at 0.42 mg/mL. However, the aqueous extract showed effects ranging from 0.42 to 1.17 mg/mL. Moringa Ag-NPs showed higher activity against the different fungal strains than the crude aqueous extract, and they ranged from 0.25 to 0.83 mg/mL. The MIC values of the Moringa crude aqueous extract ranged from 0.74 to 3.33 mg/mL. Moringa Ag-NPs and their crude aqueous extract may be utilized to boost antimicrobial attributes.
Molecular Research on Reproductive Toxicity
Authors: Scudiero Rosaria
Published: February 2023

Fertility rates in animals have shown a progressive decrease in recent decades, and reproductive toxicity is considered an important regulatory endpoint in health hazard assessment [...]
Insertion of an Amphipathic Linker in a Tetrapodal Tryptophan Derivative Leads to a Novel and Highly Potent Entry Inhibitor of Enterovirus A71 Clinical Isolates
Authors: Martí-Marí Olaia , Abdelnabi Rana , Schols Dominique , Neyts Johan , Camarasa María-José , Gago Federico , San-Félix Ana
Published: February 2023

AL-471, the leading exponent of a class of potent HIV and enterovirus A71 (EV-A71) entry inhibitors discovered in our research group, contains four l-tryptophan (Trp) units bearing an aromatic isophthalic acid directly attached to the C2 position of each indole ring. Starting from AL-471, we (i) replaced l-Trp with d-Trp, (ii) inserted a flexible linker between C2 and the isophthalic acid, and (iii) substituted a nonaromatic carboxylic acid for the terminal isophthalic acid. Truncated analogues lacking the Trp motif were also synthesized. Our findings indicate that the antiviral activity seems to be largely independent of the stereochemistry (l- or d-) of the Trp fragment and also that both the Trp unit and the distal isophthalic moiety are essential for antiviral activity. The most potent derivative, 23 (AL-534), with the C2 shortest alkyl urea linkage (three methylenes), showed subnanomolar potency against different EV-71 clinical isolates. This finding was only observed before with the early dendrimer prototype AL-385 (12 l-Trp units) but remained unprecedented for the reduced-size prototype AL-471. Molecular modeling showed the feasibility of high-affinity binding of the novel l-Trp-decorated branches of 23 (AL-534) to an alternative site on the VP1 protein that harbors significant sequence variation among EV-71 strains.
Assessment of the Impact of Physical Activity on the Musculoskeletal System in Early Degenerative Knee Joint Lesions in an Animal Model
Authors: Jarecki Jaromir , Polkowska Izabela , Kazimierczak Waldemar , Wójciak Magdalena , Sowa Ireneusz , Dresler Sławomir , Blicharski Tomasz
Published: February 2023

Osteoarthritis (OA) is one of the most prevalent diseases of the osteoarticular system. Progressive destruction of joints is accompanied by development of pathological changes in the muscle tissue, i.e., weakening, atrophy, and remodelling (sarcopenia). The aim of the present study is to assess the impact of physical activity on the musculoskeletal system in an animal model of early degenerative lesions in the knee joint. The study involved 30 male Wistar rats. The animals were allocated to three subgroups of 10 animals each. Each animal from the three subgroups received sodium iodoacetate by injection into the patellar ligament of the right knee joint, whereas saline was administered through the patellar ligament in the left knee joint. The rats in the first group were stimulated to exercise on a treadmill. The animals in the second group were allowed to lead a natural lifestyle (no treadmill stimulation). In the third group, all parts of the right hind limb muscle were injected with Clostridium botulinum toxin type A. The study demonstrated that, compared to the active rats, bone density in the immobilised rats decreased, as indicated by the densitometric assessment of the whole body and the examination of rats’ hind limbs and knee joints alone. This clearly evidenced the impact of physical activity on bone mineralisation. The weight of both fat and muscle tissues in the physically inactive rats was reduced. Additionally, the adipose tissue had higher weight in the entire right hind limbs, where monoiodoacetic acid was administered to the knee joint. The animal model clearly showed the importance of physical activity in the early stages of OA, as it slows down the process of joint destruction, bone atrophy, and muscle wasting, whereas physical inactivity contributes to progression of generalised changes in the musculoskeletal system.
Seasonal Changes in Serum Metabolites in Multiple Sclerosis Relapse
Authors: Martynova Ekaterina , Khaibullin Timur , Salafutdinov Ilnur , Markelova Maria , Laikov Alexander , Lopukhov Leonid , Liu Rongzeng , Sahay Kritika , Goyal Mehendi , Baranwal Manoj , Rizvanov Albert A , Khaiboullina Svetlana
Published: February 2023

Multiple sclerosis (MS) is a debilitating chronic disease of unknown etiology. There are limited treatment options due to an incomplete understanding of disease pathology. The disease is shown to have seasonal exacerbation of clinical symptoms. The mechanisms of such seasonal worsening of symptoms remains unknown. In this study, we applied targeted metabolomics analysis of serum samples using LC-MC/MC to determine seasonal changes in metabolites throughout the four seasons. We also analyzed seasonal serum cytokine alterations in patients with relapsed MS. For the first time, we can demonstrate seasonal changes in various metabolites in MS compared to the control. More metabolites were affected in MS in the fall season followed by spring, while summer MS was characterized by the smallest number of affected metabolites. Ceramides were activated in all seasons, suggesting their central role in the disease pathogenesis. Substantial changes in glucose metabolite levels were found in MS, indicating a potential shift to glycolysis. An increased serum level of quinolinic acid was demonstrated in winter MS. Histidine pathways were affected, suggesting their role in relapse of MS in the spring and fall. We also found that spring and fall seasons had a higher number of overlapping metabolites affected in MS. This could be explained by patients having a relapse of symptoms during these two seasons.
Ectopic Expression of PvHMA2.1 Enhances Cadmium Tolerance in Arabidopsis thaliana
Authors: Zang Hui , He Junyi , Zhang Qi , Li Xue , Wang Tingting , Bi Xiaojing , Zhang Yunwei
Published: February 2023

Cadmium (Cd) in soil inhibits plant growth and development and even harms human health through food chain transmission. Switchgrass (Panicum virgatum L.), a perennial C4 biofuel crop, is considered an ideal plant for phytoremediation due to its high efficiency in removing Cd and other heavy metals from contaminated soil. The key to understanding the mechanisms of switchgrass Cd tolerance is to identify the genes involved in Cd transport. Heavy-metal ATPases (HMAs) play pivotal roles in heavy metal transport, including Cd, in Arabidopsis thaliana and Oryza sativa, but little is known about the functions of their orthologs in switchgrass. Therefore, we identified 22 HMAs in switchgrass, which were distributed on 12 chromosomes and divided into 4 groups using a phylogenetic analysis. Then, we focused on PvHMA2.1, which is one of the orthologs of the rice Cd transporter OsHMA2. We found that PvHMA2.1 was widely expressed in roots, internodes, leaves, spikelets, and inflorescences, and was significantly induced in the shoots of switchgrass under Cd treatment. Moreover, PvHMA2.1 was found to have seven transmembrane domains and localized at the cell plasma membrane, indicating that it is a potential transporter. The ectopic expression of PvHMA2.1 alleviated the reduction in primary root length and the loss of fresh weight of Arabidopsis seedlings under Cd treatment, suggesting that PvHMA2.1 enhanced Cd tolerance in Arabidopsis. The higher levels of relative water content and chlorophyll content of the transgenic lines under Cd treatment reflected that PvHMA2.1 maintained water retention capacity and alleviated photosynthesis inhibition under Cd stress in Arabidopsis. The roots of the PvHMA2.1 ectopically expressed lines accumulated less Cd compared to the WT, while no significant differences were found in the Cd contents of the shoots between the transgenic lines and the WT under Cd treatment, suggesting that PvHMA2.1 reduced Cd absorption from the environment through the roots in Arabidopsis. Taken together, our results showed that PvHMA2.1 enhanced Cd tolerance in Arabidopsis, providing a promising target that could be engineered in switchgrass to repair Cd-contaminated soil.
Nanomicellar Formulations Loaded with Histamine and Paclitaxel as a New Strategy to Improve Chemotherapy for Breast Cancer
Authors: Nicoud Melisa B. , Ospital Ignacio A. , Táquez Delgado Mónica A. , Riedel Jennifer , Fuentes Pedro , Bernabeu Ezequiel , Rubinstein Mara R. , Lauretta Paolo , Martínez Vivot Rocío , Aguilar María de los Ángeles , Salgueiro María J. , Speisky Daniela , Moretton Marcela A. , Chiappetta Diego A. , Medina Vanina A.
Published: February 2023

Triple negative breast cancer (TNBC) is the most aggressive breast cancer subtype. Currently, paclitaxel (PTX) represents the first-line therapy for TNBC; however it presents a hydrophobic behavior and produces severe adverse effects. The aim of this work is to improve the therapeutic index of PTX through the design and characterization of novel nanomicellar polymeric formulations composed of a biocompatible copolymer Soluplus® (S), surface-decorated with glucose (GS), and co-loaded either with histamine (HA, 5 mg/mL) and/or PTX (4 mg/mL). Their micellar size, evaluated by dynamic light scattering, showed a hydrodynamic diameter between 70 and 90 nm for loaded nanoformulations with a unimodal size distribution. Cytotoxicity and apoptosis assays were performed to assess their efficacy in vitro in human MDA-MB-231 and murine 4T1 TNBC cells rendering optimal antitumor efficacy in both cell lines for the nanoformulations with both drugs. In a model of TNBC developed in BALB/c mice with 4T1 cells, we found that all loaded micellar systems reduced tumor volume and that both HA and HA-PTX-loaded SG micelles reduced tumor weight and neovascularization compared with the empty micelles. We conclude that HA-PTX co-loaded micelles in addition to HA-loaded formulations present promising potential as nano-drug delivery systems for cancer chemotherapy.
Salinity-Induced Cytosolic Alkaline Shifts in Arabidopsis Roots Require the SOS Pathway
Authors: Rombolá-Caldentey Belén , Andrés Zaida , Waadt Rainer , Quintero Francisco J. , Schumacher Karin , Pardo José M.
Published: February 2023

Plants have evolved elaborate mechanisms to sense, respond to and overcome the detrimental effects of high soil salinity. The role of calcium transients in salinity stress signaling is well established, but the physiological significance of concurrent salinity-induced changes in cytosolic pH remains largely undefined. Here, we analyzed the response of Arabidopsis roots expressing the genetically encoded ratiometric pH-sensor pHGFP fused to marker proteins for the recruitment of the sensor to the cytosolic side of the tonoplast (pHGFP-VTI11) and the plasma membrane (pHGFP-LTI6b). Salinity elicited a rapid alkalinization of cytosolic pH (pHcyt) in the meristematic and elongation zone of wild-type roots. The pH-shift near the plasma membrane preceded that at the tonoplast. In pH-maps transversal to the root axis, the epidermis and cortex had cells with a more alkaline pHcyt relative to cells in the stele in control conditions. Conversely, seedlings treated with 100 mM NaCl exhibited an increased pHcyt in cells of the vasculature relative to the external layers of the root, and this response occurred in both reporter lines. These pHcyt changes were substantially reduced in mutant roots lacking a functional SOS3/CBL4 protein, suggesting that the operation of the SOS pathway mediated the dynamics of pHcyt in response to salinity.
Solute Carrier Family 29A1 Mediates In Vitro Resistance to Azacitidine in Acute Myeloid Leukemia Cell Lines
Authors: Kutyna Monika M. , Loone Sophie , Saunders Verity A. , White Deborah L. , Kok Chung H. , Hiwase Devendra K.
Published: February 2023

Azacitidine (AZA) is commonly used hypomethylating agent for higher risk myelodysplastic syndromes and acute myeloid leukemia (AML). Although some patients achieve remission, eventually most patients fail AZA therapy. Comprehensive analysis of intracellular uptake and retention (IUR) of carbon-labeled AZA (14C-AZA), gene expression, transporter pump activity with or without inhibitors, and cytotoxicity in naïve and resistant cell lines provided insight into the mechanism of AZA resistance. AML cell lines were exposed to increasing concentrations of AZA to create resistant clones. 14C-AZA IUR was significantly lower in MOLM-13- (1.65 ± 0.08 ng vs. 5.79 ± 0.18 ng; p < 0.0001) and SKM-1- (1.10 ± 0.08 vs. 5.08 ± 0.26 ng; p < 0.0001) resistant cells compared to respective parental cells. Importantly, 14C-AZA IUR progressively reduced with downregulation of SLC29A1 expression in MOLM-13- and SKM-1-resistant cells. Furthermore, nitrobenzyl mercaptopurine riboside, an SLC29A inhibitor, reduced 14C-AZA IUR in MOLM-13 (5.79 ± 0.18 vs. 2.07 ± 0.23, p < 0.0001) and SKM-1-naive cells (5.08 ± 2.59 vs. 1.39 ± 0.19, p = 0.0002) and reduced efficacy of AZA. As the expression of cellular efflux pumps such as ABCB1 and ABCG2 did not change in AZA-resistant cells, they are unlikely contribute to AZA resistance. Therefore, the current study provides a causal link between in vitro AZA resistance and downregulation of cellular influx transporter SLC29A1.