bluuebunny/crossref_metadata_2025 huggingface dataset:
Soy and Frequent Dairy Consumption with Subsequent Equol Production Reveals Decreased Gut Health in a Cohort of Healthy Puerto Rican Women
Authors: Lacourt-Ventura Mercedes Y. , Vilanova-Cuevas Brayan , Rivera-Rodríguez Delmarie , Rosario-Acevedo Raysa , Miranda Christine , Maldonado-Martínez Gerónimo , Maysonet Johanna , Vargas Darlene , Ruiz Yelitza , Hunter-Mellado Robert , Cubano Luis A. , Dharmawardhane Suranganie , Lampe Johanna W. , Baerga-Ortiz Abel , Godoy-Vitorino Filipa , Martínez-Montemayor Michelle M.
Published: August 2021

The U.S. Hispanic female population has one of the highest breast cancer (BC) incidence and mortality rates, while BC is the leading cause of cancer death in Puerto Rican women. Certain foods may predispose to carcinogenesis. Our previous studies indicate that consuming combined soy isoflavones (genistein, daidzein, and glycitein) promotes tumor metastasis possibly through increased protein synthesis activated by equol, a secondary dietary metabolite. Equol is a bacterial metabolite produced in about 20–60% of the population that harbor and exhibit specific gut microbiota capable of producing it from daidzein. The aim of the current study was to investigate the prevalence of equol production in Puerto Rican women and identify the equol producing microbiota in this understudied population. Herein, we conducted a cross-sectional characterization of equol production in a clinically based sample of eighty healthy 25–50 year old Puerto Rican women. Urine samples were collected and evaluated by GCMS for the presence of soy isoflavones and metabolites to determine the ratio of equol producers to equol non-producers. Furthermore, fecal samples were collected for gut microbiota characterization on a subset of women using next generation sequencing (NGS). We report that 25% of the participants were classified as equol producers. Importantly, the gut microbiota from equol non-producers demonstrated a higher diversity. Our results suggest that healthy women with soy and high dairy consumption with subsequent equol production may result in gut dysbiosis by having reduced quantities (diversity) of healthy bacterial biomarkers, which might be associated to increased diseased outcomes (e.g., cancer, and other diseases).
Perceived Importance of Public Health Risks in Greece: A Nationwide Survey of the Adult Population
Authors: Tzortzi Anna , Kapetanstrataki Melpo , Rachiotis Georgios , Evangelopoulou Vaso , Leventou Eleni , Behrakis Panagiotis
Published: August 2021

The current study is the first to examine the perceptions of the Greek public towards selected health risks and prioritize perceived risk importance and the needs to be addressed for public health promotion. Participants were asked to consider the individual importance of selected risks and the top three most important risks. Data collection took place on February 2020 in a representative sample of the adult Greek population. Differences between groups were assessed with Chi-square tests. Logistic regression models were used to identify perceptions based on participants’ characteristics. Analysis was conducted in Stata 14, and 1976 adults participated in the survey: 48% male and 52% female. Road accidents, cancer and air pollution and environmental protection were considered the top three most important public health risks. Differences were observed between sexes; females were more concerned regarding the importance of the examined public health risks, and between age groups, younger ages considered STDs and contraception more important than older ages. Finally, non-smokers considered exercise and smoking to be more important than smokers. This is the first study to present and grade the public’s perceptions on the importance of public health risks in Greece. Our study’s prioritization of health risks could aid health authorities in improving and promoting the overall public health in Greece.
Older Adults with Hypertension: Prevalence of Falls and Their Associated Factors
Authors: Abu Bakar Atirah Az-Zahra , Abdul Kadir Azidah , Idris Nur Suhaila , Mohd Nawi Siti Nurbaya
Published: August 2021

Falls are prominent health issues among older adults. Among hypertensive older adults, falls may have a detrimental effect on their health and wellbeing. The purpose of this study is to determine the prevalence of falls among hypertensive older adults and to identify the associated factors that contribute to their falls. This was a cross-sectional study conducted among two hundred and sixty-nine hypertensive older adults who were selected via systematic random sampling in two primary health clinics in Kuala Terengganu, Malaysia. Data on their socio-demographic details, their history of falls, medication history and clinical characteristics were collected. Balance and gait were assessed using the Performance Oriented Mobility Assessment (POMA). It was found that 32.2% of participants reported falls within a year. Polypharmacy (adjusted OR 2.513, 95% CI 1.339, 4.718) and diuretics (adjusted OR 2.803, 95% CI 1.418, 5.544) were associated with an increased risk of falls. Meanwhile, a higher POMA score (adjusted OR 0.940, 95% CI 0.886, 0.996) and the number of antihypertensives (adjusted OR 0.473, 95% CI 0.319, 0.700) were associated with a low incidence of falling among hypertensive older adults. Falls are common among hypertensive older adults. Older adults who are taking diuretics and have a polypharmacy treatment plan have a higher incidence of falls. However, older adults taking a higher number of anti-hypertensive medications specifically were not associated with an increased prevalence of falls.
Sagittal Integral Morphotype of Competitive Amateur Athletes and Its Potential Relation with Recurrent Low Back Pain
Authors: Cejudo Antonio , Centenera-Centenera Josep María , Santonja-Medina Fernando
Published: August 2021

Athletes have higher thoracic and lumbar curvature in standing than the reference values of the non-athletic population. The sagittal integral morphotype method (SIM) assessment has not previously been applied to competitive amateur athletes (CAA). The propose of the present study was to determine the SIM of CAA treated at a sports-medicine center and to identify spinal misalignments associated with recurrent low back pain (LBP). An observational analysis was developed to describe the SIM in 94 CAA. The thoracic and lumbar curvatures of the CAA were measured in standing, sitting, and trunk forward flexion. Association analysis (Pearson’s chi-square and Cramér’s V tests) was then performed to identify the SIM misalignments associated with LBP. Effect size was analyzed based on Hedges’ g. The most common thoracic SIMs in CAA were total hyperkyphosis (male = 59.02%; female = 42.42%) and static hyperkyphosis (male = 11.48%; female = 6.06%). Hyperlordotic attitude (female = 30.30%; male = 4.92%), static-functional hyperkyphosis (male = 16.39%; female = 3.03%), and structured hyperlordosis (female = 21.21%; male = 1.64%) were the most common lumbar SIMs. Hyperlordotic attitude, static functional lumbar hyperkyphosis, and structured hyperlordosis were associated with LBP in male and female athletes.
Good Character at College: The Combined Role of Second-Order Character Strength Factors and Phronesis Motivation in Undergraduate Academic Outcomes
Authors: Villacís Jorge L. , de la Fuente Jesús , Naval Concepción
Published: August 2021

A renewed interest in the study of character and virtue has recently emerged in the fields of Education and Psychology. The latest research has confirmed the association between virtuous consistent behaviours and academic positive outcomes. However, the motivational dimension of character (the intentions underlying the patterns of observed behaviours) has received little attention. This research aims to extend the knowledge on this topic by examining the predictive relationships between the behavioural and motivational dimensions of character, with reference to academic engagement, career self-doubt and performance of Spanish university students. A total of 183 undergraduates aged 18–30 (142 of whom were women) from the north of Spain completed specific parts of self-report questionnaires, including the Values in Action VIA-72, a Spanish translated and validated version of the Moral Self-Relevance Measure MSR, and the Utrecht Work Engagement Student Scale UWES-S9. The collected data were analysed using Structural Equation Modelling. The behavioural dimension of character (character strength factors of caring, self-control and inquisitiveness) showed positive associations with academic engagement and performance. The motivational dimension of character (phronesis motivation), was negatively related to career self-doubt. For the first time, the present study has provided support for the contribution of both dimensions of character to undergraduate academic outcomes.
Development of a Scale Measuring Emotional Catharsis through Illness Narratives
Authors: Liao Hung-Chang , Wang Ya-huei
Published: August 2021

Objective: This study intended to construct a scale measuring the catharsis effect on medical professionals or students through illness narratives (ECS-IN). Methods: After a systematic literature review and panel discussion, the researchers conducted a pilot study with a sample of seven hundred and eighty-two randomly selected healthcare students and providers in Taiwan to examine psychometric properties using exploratory factor analysis (EFA) and confirmatory factor analysis (CFA) for item derivation and factor extraction. The researchers also tested the validities and reliabilities of the ECS-IN scale to confirm its feasibility. Results: the EFA yielded 29 items and three factors: “emotional identification as self-healing” (12 items; 55.500% of variance explained), “emotional release for compensation” (10 items; 7.465% of variance explained), and “emotional adjustment for intellectual growth” (7 items; 4.839% of variance explained). The CFA yielded an 18-item, three-factor model with satisfactory fit to the data, where the χ2/df ratio = 1.090, Tucker–Lewis index (TLI) = 0.996, comparative fit index (CFI) = 0.997, and root mean square of approximation (RMSEA) = 0.020. The convergent validity and discriminant validities also demonstrated the feasibility of the ECS-IN scale. For the first version of the ECS-IN scale (29 items), the Cronbach’s alphas for the three factors and the overall scale were in the range between 0.912 and 0.971; for the reduced version of the scale (18 items), the Cronbach’s alphas and composite reliabilities were in the range of 0.888–0.946 and 0.890–0.968. Conclusion: The findings proved that the ECS-IN could be a reliable and valid instrument to assess participants’ emotional catharsis through illness narratives.
Application of Multivariate Statistical Techniques and Water Quality Index for the Assessment of Water Quality and Apportionment of Pollution Sources in the Yeongsan River, South Korea
Authors: Mamun Md , An Kwang-Guk
Published: August 2021

This study assessed spatial and temporal variations of water quality to identify and quantify possible pollution sources affecting the Yeongsan River using multivariate statistical techniques (MSTs) and water quality index (WQI) values. A 15 year dataset of 11 water quality variables was used, covering 16 monitoring sites. The nutrient regime, organic matter, suspended solids, ionic contents, algal growth, and total coliform bacteria (TCB) were affected by the summer monsoon and the construction of weirs. Regression analysis showed that the algal growth was more highly regulated by total phosphorus (TP; R2 = 0.37) than total nitrogen (TN, R2 = 0.25) and TN/TP (R2 = 0.01) ratios in the river after weir construction and indicated that the river is a P-limited system. After constructing the weirs, the mean TN/TP ratio in the river was about 40, meaning it is a P-limited system. Cluster analysis was used to classify the sampling sites into highly, moderately, and less polluted sites based on water quality features. Stepwise discriminant analysis showed that pH, dissolved oxygen (DO), TN, biological oxygen demand (BOD), chemical oxygen demand (COD), chlorophyll-a (CHL-a), and TCB are the spatially discriminating parameters, while pH, water temperature, DO, electrical conductivity, total suspended solids, and COD are the most significant for discriminating among the three seasons. The Pearson network analysis showed that nutrients flow with organic matter in the river, while CHL-a showed the highest correlation with COD (r = 0.85), followed by TP (r = 0.49) and TN (r = 0.49). Average WQI values ranged from 55 to 141, indicating poor to unsuitable water quality in the river. The Mann–Kendall test showed increasing trends in COD and CHL-a but decreasing trends for TP, TN, and BOD due to impoundment effects. The principal component analysis combined with factor analysis and positive matrix factorization (PMF) showed that two sewage treatment plants, agricultural activities, and livestock farming adversely impacted river water quality. The PMF model returned greater R2 values for BOD (0.92), COD (0.87), TP (0.93), TN (0.91), CHL-a (0.93), and TCB (0.83), indicating reliable apportionment results. Our results suggest that MSTs and WQI can be effectively used for the simple interpretation of large-scale datasets to determine pollution sources and their spatiotemporal variations. The outcomes of our study may aid policymakers in managing the Yeongsan River.
Extracellular Vesicles as Biological Indicators and Potential Sources of Autologous Therapeutics in Osteoarthritis
Authors: Zhang Xin , Huebner Janet L. , Kraus Virginia Byers
Published: August 2021

Along with cytokines, extracellular vesicles (EVs) released by immune cells in the joint contribute to osteoarthritis (OA) pathogenesis. By high-resolution flow cytometry, we characterized 18 surface markers and 4 proinflammatory cytokines carried by EVs of various sizes in plasma and synovial fluid (SF) from individuals with knee OA, with a primary focus on immune cells that play a major role in OA pathogenesis. By multiplex immunoassay, we also measured concentrations of cytokines within (endo) and outside (exo) EVs. EVs carrying HLA-DR, -DP and -DQ were the most enriched subpopulations in SF relative to plasma (25–50-fold higher depending on size), suggesting a major contribution to the SF EV pool from infiltrating immune cells in OA joints. In contrast, the CD34+ medium and small EVs, reflecting hematopoietic stem cells, progenitor cells, and endothelial cells, were the most significantly enriched subpopulations in plasma relative to SF (7.3- and 7.7-fold higher). Ratios of EVs derived from neutrophils and lymphocytes were highly correlated between SF and plasma, indicating that plasma EVs could reflect OA severity and serve as systemic biomarkers of OA joint pathogenesis. Select subsets of plasma EVs might also provide next generation autologous biological products for intra-articular therapy of OA joints.
Development of a Bispecific Antibody-Based Platform for Retargeting of Capsid Modified AAV Vectors
Authors: Kuklik Juliane , Michelfelder Stefan , Schiele Felix , Kreuz Sebastian , Lamla Thorsten , Müller Philipp , Park John E.
Published: August 2021

A major limiting factor for systemically delivered gene therapies is the lack of novel tissue specific AAV (Adeno-associated virus) derived vectors. Bispecific antibodies can be used to redirect AAVs to specific target receptors. Here, we demonstrate that the insertion of a short linear epitope “2E3” derived from human proprotein-convertase subtilisin/kexin type 9 (PCSK9) into different surface loops of the VP capsid proteins can be used for AAV de-targeting from its natural receptor(s), combined with a bispecific antibody-mediated retargeting. We chose to target a set of distinct disease relevant membrane proteins—fibroblast activation protein (FAP), which is upregulated on activated fibroblasts within the tumor stroma and in fibrotic tissues, as well as programmed death-ligand 1 (PD-L1), which is strongly upregulated in many cancers. Upon incubation with a bispecific antibody recognizing the 2E3 epitope and FAP or PD-L1, the bispecific antibody/rAAV complex was able to selectively transduce receptor positive cells. In summary, we developed a novel, rationally designed vector retargeting platform that can target AAVs to a new set of cellular receptors in a modular fashion. This versatile platform may serve as a valuable tool to investigate the role of disease relevant cell types and basis for novel gene therapy approaches.
Zebrafish Motile Cilia as a Model for Primary Ciliary Dyskinesia
Authors: Pinto Andreia L. , Rasteiro Margarida , Bota Catarina , Pestana Sara , Sampaio Pedro , Hogg Claire , Burgoyne Thomas , Lopes Susana S.
Published: August 2021

Zebrafish is a vertebrate teleost widely used in many areas of research. As embryos, they develop quickly and provide unique opportunities for research studies owing to their transparency for at least 48 h post fertilization. Zebrafish have many ciliated organs that include primary cilia as well as motile cilia. Using zebrafish as an animal model helps to better understand human diseases such as Primary Ciliary Dyskinesia (PCD), an autosomal recessive disorder that affects cilia motility, currently associated with more than 50 genes. The aim of this study was to validate zebrafish motile cilia, both in mono and multiciliated cells, as organelles for PCD research. For this purpose, we obtained systematic high-resolution data in both the olfactory pit (OP) and the left–right organizer (LRO), a superficial organ and a deep organ embedded in the tail of the embryo, respectively. For the analysis of their axonemal ciliary structure, we used conventional transmission electron microscopy (TEM) and electron tomography (ET). We characterised the wild-type OP cilia and showed, for the first time in zebrafish, the presence of motile cilia (9 + 2) in the periphery of the pit and the presence of immotile cilia (still 9 + 2), with absent outer dynein arms, in the centre of the pit. In addition, we reported that a central pair of microtubules in the LRO motile cilia is common in zebrafish, contrary to mouse embryos, but it is not observed in all LRO cilia from the same embryo. We further showed that the outer dynein arms of the microtubular doublet of both the OP and LRO cilia are structurally similar in dimensions to the human respiratory cilia at the resolution of TEM and ET. We conclude that zebrafish is a good model organism for PCD research but investigators need to be aware of the specific physical differences to correctly interpret their results.
G2-S16 Polyanionic Carbosilane Dendrimer Can Reduce HIV-1 Reservoir Formation by Inhibiting Macrophage Cell to Cell Transmission
Authors: Relaño-Rodríguez Ignacio , Espinar-Buitrago María de la Sierra , Martín-Cañadilla Vanessa , Gómez-Ramírez Rafael , Muñoz-Fernández María Ángeles
Published: August 2021

Human immunodeficiency virus (HIV-1) is still a major problem, not only in developing countries but is also re-emerging in several developed countries, thus the development of new compounds able to inhibit the virus, either for prophylaxis or treatment, is still needed. Nanotechnology has provided the science community with several new tools for biomedical applications. G2-S16 is a polyanionic carbosilane dendrimer capable of inhibiting HIV-1 in vitro and in vivo by interacting directly with viral particles. One of the main barriers for HIV-1 eradication is the reservoirs created in primoinfection. These reservoirs, mainly in T cells, are untargetable by actual drugs or immune system. Thus, one approach is inhibiting HIV-1 from reaching these reservoir cells. In this context, macrophages play a main role as they can deliver viral particles to T cells establishing reservoirs. We showed that G2-S16 dendrimer is capable of inhibiting the infection from infected macrophages to healthy T CD4/CD8 lymphocytes by eliminating HIV-1 infectivity inside macrophages, so they are not able to carry infectious particles to other body locations, thus preventing the reservoirs from forming.
Necrostatin-1 Supplementation to Islet Tissue Culture Enhances the In-Vitro Development and Graft Function of Young Porcine Islets
Authors: Lau Hien , Li Shiri , Corrales Nicole , Rodriguez Samuel , Mohammadi Mohammadreza , Alexander Michael , de Vos Paul , Lakey Jonathan
Published: August 2021

Pre-weaned porcine islets (PPIs) represent an unlimited source for islet transplantation but are functionally immature. We previously showed that necrostatin-1 (Nec-1) immediately after islet isolation enhanced the in vitro development of PPIs. Here, we examined the impact of Nec-1 on the in vivo function of PPIs after transplantation in diabetic mice. PPIs were isolated from pancreata of 8–15-day-old, pre-weaned pigs and cultured in media alone, or supplemented with Nec-1 (100 µM) on day 0 or on day 3 of culture (n = 5 for each group). On day 7, islet recovery, viability, oxygen consumption rate, insulin content, cellular composition, insulin secretion capacity, and transplant outcomes were evaluated. While islet viability and oxygen consumption rate remained high throughout 7-day tissue culture, Nec-1 supplementation on day 3 significantly improved islet recovery, insulin content, endocrine composition, GLUT2 expression, differentiation potential, proliferation capacity of endocrine cells, and insulin secretion. Adding Nec-1 on day 3 of tissue culture enhanced the islet recovery, proportion of delta cells, beta-cell differentiation and proliferation, and stimulation index. In vivo, this leads to shorter times to normoglycemia, better glycemic control, and higher circulating insulin. Our findings identify the novel time-dependent effects of Nec-1 supplementation on porcine islet quantity and quality prior to transplantation.
Molecular Pathogenesis and Peripheral Monitoring of Adult Fragile X-Associated Syndromes
Authors: Valor Luis M. , Morales Jorge C. , Hervás-Corpión Irati , Marín Rosario
Published: August 2021

Abnormal trinucleotide expansions cause rare disorders that compromise quality of life and, in some cases, lifespan. In particular, the expansions of the CGG-repeats stretch at the 5’-UTR of the Fragile X Mental Retardation 1 (FMR1) gene have pleiotropic effects that lead to a variety of Fragile X-associated syndromes: the neurodevelopmental Fragile X syndrome (FXS) in children, the late-onset neurodegenerative disorder Fragile X-associated tremor-ataxia syndrome (FXTAS) that mainly affects adult men, the Fragile X-associated primary ovarian insufficiency (FXPOI) in adult women, and a variety of psychiatric and affective disorders that are under the term of Fragile X-associated neuropsychiatric disorders (FXAND). In this review, we will describe the pathological mechanisms of the adult “gain-of-function” syndromes that are mainly caused by the toxic actions of CGG RNA and FMRpolyG peptide. There have been intensive attempts to identify reliable peripheral biomarkers to assess disease progression and onset of specific pathological traits. Mitochondrial dysfunction, altered miRNA expression, endocrine system failure, and impairment of the GABAergic transmission are some of the affectations that are susceptible to be tracked using peripheral blood for monitoring of the motor, cognitive, psychiatric and reproductive impairment of the CGG-expansion carriers. We provided some illustrative examples from our own cohort. Understanding the association between molecular pathogenesis and biomarkers dynamics will improve effective prognosis and clinical management of CGG-expansion carriers.
Transcriptomic Analysis Reveals Key Genes Involved in Oil and Linoleic Acid Biosynthesis during Artemisia sphaerocephala Seed Development
Authors: Nan Shuzhen , Zhang Lijing , Hu Xiaowei , Miao Xiumei , Han Xiaoxu , Fu Hua
Published: August 2021

Artemisia sphaerocephala seeds are rich in polysaccharides and linoleic acid (C18:2), which have been widely used as traditional medicine and to improve food quality. The accumulation patterns and molecular regulatory mechanisms of polysaccharides during A. sphaerocephala seed development have been studied. However, the related research on seed oil and C18:2 remain unclear. For this study, A. sphaerocephala seeds at seven different development stages at 10, 20, 30, 40, 50, 60, and 70 days after flowering (designated as S1~S7), respectively, were employed as experimental samples, the accumulation patterns of oil and fatty acids (FA) and the underlying molecular regulatory mechanisms were analyzed. The results revealed that oil content increased from 10.1% to 20.0% in the early stages of seed development (S1~S2), and up to 32.0% in mature seeds, of which C18:2 accounted for 80.6% of the total FA. FA and triacylglycerol biosynthesis-related genes jointly involved in the rapid accumulation of oil in S1~S2. Weighted gene co-expression network analysis showed that transcription factors FUS3 and bHLH played a critical role in the seed oil biosynthesis. The perfect harmonization of the high expression of FAD2 with the extremely low expression of FAD3 regulated the accumulation of C18:2. This study uncovered the gene involved in oil biosynthesis and molecular regulatory mechanisms of high C18:2 accumulation in A. sphaerocephala seeds; thus, advancing research into unsaturated fatty acid metabolism in plants while generating valuable genetic resources for optimal C18:2 breeding.
Melatonin and Glycine Reduce Uterus Ischemia/Reperfusion Injury in a Rat Model of Warm Ischemia
Authors: Zitkute Viktorija , Kvietkauskas Mindaugas , Maskoliunaite Vygante , Leber Bettina , Ramasauskaite Diana , Strupas Kestutis , Stiegler Philipp , Schemmer Peter
Published: August 2021

Ischemia/reperfusion injury (IRI) remains a significant problem to be solved in uterus transplantation (UTx). Melatonin and glycine have been shown to possess direct cytoprotective activities, mainly due to their antioxidative and anti-inflammatory properties. The aim of this study was to investigate the protective effects of melatonin and glycine and their combination on IRI in a rat model of warm ischemia. In this study, Sprague-Dawley rats were assigned to eight groups, including sham and IRI (n = 80). Melatonin and glycine alone or their combination were administered prior to 1 h of uterus ischemia followed by 1 h of reperfusion. Melatonin (50 mg/kg) was administered via gavage 2 h before IRI and glycine in an enriched diet for 5 days prior to intervention. Uterus IRI was estimated by histology, including immunohistochemistry, and biochemical tissue analyses. Histology revealed that uterus IRI was significantly attenuated by pretreatment with melatonin (p = 0.019) and glycine (p = 0.044) alone as well as their combination (p = 0.003). Uterus IRI led to increased myeloperoxidase expression, which was significantly reduced by melatonin (p = 0.004), glycine (p < 0.001) or their combination (p < 0.001). The decline in superoxide dismutase activity was significantly reduced in the melatonin (p = 0.027), glycine (p = 0.038) and combined treatment groups (p = 0.015) when compared to the IRI control group. In conclusion, melatonin, glycine and their combination significantly reduced oxidative stress-induced cell damage after IRI in a small animal warm ischemia model, and, therefore, clinical studies are required to evaluate the protective effects of these well-characterized substances in uterus IRI.
Bovine Satellite Cells Isolated after 2 and 5 Days of Tissue Storage Maintain the Proliferative and Myogenic Capacity Needed for Cultured Meat Production
Authors: Skrivergaard Stig , Rasmussen Martin Krøyer , Therkildsen Margrethe , Young Jette Feveile
Published: August 2021

Cultured meat is an emerging alternative food technology which aims to deliver a more ethical, sustainable, and healthy muscle-tissue-derived food item compared to conventional meat. As start-up companies are rapidly forming and accelerating this technology, many aspects of this multi-faceted science have still not been investigated in academia. In this study, we investigated if bovine satellite cells with the ability to proliferate and undergo myogenic differentiation could be isolated after extended tissue storage, for the purpose of increasing the practicality for cultured meat production. Proliferation of bovine satellite cells isolated on the day of arrival or after 2 and 5 days of tissue storage were analyzed by metabolic and DNA-based assays, while their myogenic characteristics were investigated using RT-qPCR and immunofluorescence. Extended tissue storage up to 5 days did not negatively affect proliferation nor the ability to undergo fusion and create myosin heavy chain-positive myotubes. The expression patterns of myogenic and muscle-specific genes were also not affected after tissue storage. In fact, the data indicated a positive trend in terms of myogenic potential after tissue storage, although it was non-significant. These results suggest that the timeframe of which viable myogenic satellite cells can be isolated and used for cultured meat production can be greatly extended by proper tissue storage.
High Concentration of an ISS-N1-Targeting Antisense Oligonucleotide Causes Massive Perturbation of the Transcriptome
Authors: Ottesen Eric William , Luo Diou , Singh Natalia Nikolaevna , Singh Ravindra Narayan
Published: August 2021

Intronic splicing silencer N1 (ISS-N1) located within Survival Motor Neuron 2 (SMN2) intron 7 is the target of a therapeutic antisense oligonucleotide (ASO), nusinersen (Spinraza), which is currently being used for the treatment of spinal muscular atrophy (SMA), a leading genetic disease associated with infant mortality. The discovery of ISS-N1 as a promising therapeutic target was enabled in part by Anti-N1, a 20-mer ASO that restored SMN2 exon 7 inclusion by annealing to ISS-N1. Here, we analyzed the transcriptome of SMA patient cells treated with 100 nM of Anti-N1 for 30 h. Such concentrations are routinely used to demonstrate the efficacy of an ASO. While 100 nM of Anti-N1 substantially stimulated SMN2 exon 7 inclusion, it also caused massive perturbations in the transcriptome and triggered widespread aberrant splicing, affecting expression of essential genes associated with multiple cellular processes such as transcription, splicing, translation, cell signaling, cell cycle, macromolecular trafficking, cytoskeletal dynamics, and innate immunity. We validated our findings with quantitative and semiquantitative PCR of 39 candidate genes associated with diverse pathways. We also showed a substantial reduction in off-target effects with shorter ISS-N1-targeting ASOs. Our findings are significant for implementing better ASO design and dosing regimens of ASO-based drugs.
Hyaluronan/Diethylaminoethyl Chitosan Polyelectrolyte Complexes as Carriers for Improved Colistin Delivery
Authors: Dubashynskaya Natallia V. , Raik Sergei V. , Dubrovskii Yaroslav A. , Demyanova Elena V. , Shcherbakova Elena S. , Poshina Daria N. , Shasherina Anna Y. , Anufrikov Yuri A. , Skorik Yury A.
Published: August 2021

Improving the therapeutic characteristics of antibiotics is an effective strategy for controlling the growth of multidrug-resistant Gram-negative microorganisms. The purpose of this study was to develop a colistin (CT) delivery system based on hyaluronic acid (HA) and the water-soluble cationic chitosan derivative, diethylaminoethyl chitosan (DEAECS). The CT delivery system was a polyelectrolyte complex (PEC) obtained by interpolymeric interactions between the HA polyanion and the DEAECS polycation, with simultaneous inclusion of positively charged CT molecules into the resulting complex. The developed PEC had a hydrodynamic diameter of 210–250 nm and a negative surface charge (ζ-potential = −19 mV); the encapsulation and loading efficiencies were 100 and 16.7%, respectively. The developed CT delivery systems were characterized by modified release (30–40% and 85–90% of CT released in 15 and 60 min, respectively) compared to pure CT (100% CT released in 15 min). In vitro experiments showed that the encapsulation of CT in polysaccharide carriers did not reduce its antimicrobial activity, as the minimum inhibitory concentrations against Pseudomonas aeruginosa of both encapsulated CT and pure CT were 1 μg/mL.
Chilling Requirement Validation and Physiological and Molecular Responses of the Bud Endodormancy Release in Paeonia lactiflora ‘Meiju’
Authors: Zhang Runlong , Wang Xiaobin , Shi Xiaohua , Shao Lingmei , Xu Tong , Xia Yiping , Li Danqing , Zhang Jiaping
Published: August 2021

The introduction of herbaceous peony (Paeonia lactiflora Pall.) in low-latitude areas is of great significance to expand the landscape application of this world-famous ornamental. With the hazards of climate warming, warm winters occurs frequently, which makes many excellent northern herbaceous peony cultivars unable to meet their chilling requirements (CR) and leads to their poor growth and flowering in southern China. Exploring the endodormancy release mechanism of underground buds is crucial for improving low-CR cultivar screening and breeding. A systematic study was conducted on P. lactiflora ‘Meiju’, a screened cultivar with a typical low-CR trait introduced from northern China, at the morphological, physiological and molecular levels. The CR value of ‘Meiju’ was further verified as 677.5 CUs based on the UT model and morphological observation. As a kind of signal transducer, reactive oxygen species (ROS) released a signal to enter dormancy, which led to corresponding changes in carbohydrate and hormone metabolism in buds, thus promoting underground buds to acquire strong cold resistance and enter endodormancy. The expression of important genes related to ABA metabolism, such as NCED3, PP2C, CBF4 and ABF2, reached peaks at the critical stage of endodormancy release (9 January) and then decreased rapidly; the expression of the GA2ox8 gene related to GA synthesis increased significantly in the early stage of endodormancy release and decreased rapidly after the release of ecodormancy (23 January). Cytological observation showed that the period when the sugar and starch contents decreased and the ABA/GA ratio decreased was when ‘Meiju’ bud endodormancy was released. This study reveals the endodormancy regulation mechanism of ‘Meiju’ buds with the low-CR trait, which lays a theoretical foundation for breeding new herbaceous peony cultivars with the low-CR trait.
LncRNA MIR31HG Drives Oncogenicity by Inhibiting the Limb-Bud and Heart Development Gene (LBH) during Oral Carcinoma
Authors: Chang Kuo-Wei , Hung Wan-Wen , Chou Chung-Hsien , Tu Hsi-Feng , Chang Shi-Rou , Liu Ying-Chieh , Liu Chung-Ji , Lin Shu-Chun
Published: August 2021

The miR-31 host gene (MIR31HG) encodes a long non-coding RNA (LncRNA) that harbors miR-31 in its intron 2; miR-31 promotes malignant neoplastic progression. Overexpression of MIR31HG and of miR-31 occurs during oral squamous cell carcinoma (OSCC). However, the downstream effectors modulated by MIR31HG during OSCC pathogenesis remain unclear. The present study identifies up-regulation of MIR31HG expression during the potentially premalignant disorder stage of oral carcinogenesis. The potential of MIR31HG to enhance oncogenicity and to activate Wnt and FAK was identified when there was exogenous MIR31HG expression in OSCC cells. Furthermore, OSCC cell subclones with MIR31HG deleted were established using a Crispr/Cas9 strategy. RNA sequencing data obtained from cells expressing MIR31HG, cells with MIR31HG deleted and cells with miR-31 deleted identified 17 candidate genes that seem to be modulated by MIR31HG in OSCC cells. A TCGA database algorithm pinpointed MMP1, BMP2 and Limb-Bud and Heart development (LBH) as effector genes controlled by MIR31HG during OSCC. Exogenous LBH expression decreases tumor cell invasiveness, while knockdown of LBH reverses the oncogenic suppression present in MIR31HG deletion subclones. The study provides novel insights demonstrating the contribution of the MIR31HG-LBH cascade to oral carcinogenesis.
Photodynamic Inactivation of an Endodontic Bacteria Using Diode Laser and Indocyanine Green-Loaded Nanosphere
Authors: Higuchi Naoya , Hayashi Jun-ichiro , Fujita Masanori , Iwamura Yuki , Sasaki Yasuyuki , Goto Ryoma , Ohno Tasuku , Nishida Eisaku , Yamamoto Genta , Kikuchi Takeshi , Mitani Akio , Fukuda Mitsuo
Published: August 2021

Apical periodontitis, an inflammatory lesion causing bone resorption around the apex of teeth, is treated by eradicating infectious bacteria from the root canal. However, it has a high recurrence rate and often requires retreatment. We investigated the bactericidal effect of antimicrobial photodynamic therapy (aPDT)/photodynamic antimicrobial chemotherapy (PACT) using indocyanine green (ICG)-loaded nanospheres coated with chitosan and a diode laser on a biofilm of Enterococcus faecalis, a pathogen of refractory apical periodontitis. Biofilm of E. faecalis was cultured in a porcine infected root canal model. ICG solution was injected into the root canal, which was then irradiated with a laser (810 nm wavelength) from outside the root canal. The bactericidal effect was evaluated by colony counts and scanning electron microscopy. The result of the colony counts showed a maximum 1.89 log reduction after irradiation at 2.1 W for 5 min. The temperature rise during aPDT/PACT was confirmed to be within a safe range. Furthermore, the light energy transmittance through the root was at a peak approximately 1 min after the start of irradiation, indicating that most of the ICG in the root canal was consumed. This study shows that aPDT/PACT can suppress E. faecalis in infected root canals with high efficiency.
An IoT-Based Mobile System for Safety Monitoring of Lone Workers
Authors: Battistoni Pietro , Sebillo Monica , Vitiello Giuliana
Published: August 2021

The European Agency for Safety and Health at Work considers Smart Personal Protective Equipment as “Intelligent Protection For The Future”. It mainly consists of electronic components that collect data about their use, the workers who wear them, and the working environment. This paper proposes a distributed solution of Smart Personal Protective Equipment for the safety monitoring of Lone Workers by adopting low-cost electronic devices. In addition to the same hazards as anyone else, Lone Workers need additional and specific systems due to the higher risk they run on a work site. To this end, the Edge-Computing paradigm can be adopted to deploy an architecture embedding wearable devices, which alerts safety managers when workers do not wear the prescribed Personal Protective Equipment and supports a fast rescue when a worker seeks help or an accidental fall is automatically detected. The proposed system is a work-in-progress which provides an architecture design to accommodate different requirements, namely the deployment difficulties at temporary and large working sites, the maintenance and connectivity recurring cost issues, the respect for the workers’ privacy, and the simplicity of use for workers and their supervisors.
Utility of the SARC-F Questionnaire for Sarcopenia Screening in Patients with Chronic Liver Disease: A Multicenter Cross-Sectional Study in Japan
Authors: Hanai Tatsunori , Hiraoka Atsushi , Shiraki Makoto , Sugimoto Ryosuke , Taniki Nobuhito , Hiramatsu Akira , Nakamoto Nobuhiro , Iwasa Motoh , Chayama Kazuaki , Shimizu Masahito
Published: August 2021

Diagnosing sarcopenia is challenging. This multicenter cross-sectional study aimed to evaluate the utility of the SARC-F score system for identifying sarcopenia in patients with chronic liver disease (CLD). We enrolled 717 patients from five participating centers who completed the SARC-F between November 2019 and March 2021. Sarcopenia was diagnosed based on the Japan Society of Hepatology Working Group on Sarcopenia in Liver Disease Consensus. Muscle strength was estimated using a grip dynamometer, and muscle mass was assessed using computed tomography or bioelectrical impedance analysis. The association between SARC-F and sarcopenia was analyzed using a logistic regression model. The optimal SARC-F cutoff value for identifying sarcopenia was determined using receiver operating characteristic (ROC) curve analysis. Of the 676 eligible patients, 15% were diagnosed with sarcopenia. The SARC-F distribution was 0 points in 63% of patients, 1 point in 17%, 2 points in 7%, 3 points in 4%, and ≥4 points in 8%. The SARC-F items of “Strength” (odds ratio (OR), 1.98; 95% confidence interval (CI), 1.03–3.80) and “Falls” (OR, 2.44; 95% CI, 1.48–4.03) were significantly associated with sarcopenia. The SARC-F value of 1 point showed a higher discriminative ability for identifying sarcopenia than the 4 points that are conventionally used (p < 0.001), with an area under the ROC curve of 0.68, sensitivity of 0.65, specificity of 0.68, positive predictive value of 0.27, and negative predictive value of 0.92. SARC-F is useful for identifying patients with CLD who are at risk of sarcopenia.
Risk of Endometrial Cancer in Women with Diabetes: A Population-Based Retrospective Cohort Study
Authors: Zabuliene Lina , Kaceniene Augustė , Steponaviciene Laura , Linkeviciute-Ulinskiene Donata , Stukas Rimantas , Arlauskas Rokas , Vanseviciute-Petkeviciene Rasa , Smailyte Giedre
Published: August 2021

The aim of this study was to examine the association between type 2 diabetes (T2DM), use of glucose-lowering medications and endometrial cancer (EC) risk. Methods: The risk of EC incidence among women with T2DM in Lithuania was assessed using a retrospective cohort study design. Female patients who were registered with T2DM between 1 January 2000 and 31 December 2012 were identified in the National Health Insurance Fund database. EC cases (ICD-10 code C54) were identified from the Lithuanian Cancer Registry. Standardized incidence ratios (SIRs) were calculated by dividing the observed numbers of EC among patients with T2DM by the expected number of EC, calculated using national rates. Results: A total of 77,708 diabetic women were included in the analysis, and 995 cases of EC were identified. A significantly increased EC risk in diabetic women was found as compared to the general population (SIR = 1.69, 95% CI 1.59–1.80). The greatest EC risk was found among younger patients at T2DM diagnosis, and the risk declined gradually with increasing age but persisted in being significantly increased among all age groups. The risk for EC increased with increasing duration of diabetes, and the highest EC risk was observed more than 10 years after T2DM diagnosis. A significantly higher EC risk than expected from the general population was found in all patient groups by glucose-lowering medication combinations. The lowest EC risk was observed in diabetic women who were users of “oral only” (without metformin) (SIR = 1.42, 95% CI 1.10–1.83) and “metformin only” (SIR = 1.69, 95% CI 1.49–1.92) medications. A two times greater EC risk was observed among the remaining glucose-lowering medication categories. In contrast, use of insulin only was not related to a higher EC incidence risk (SIR = 0.45, 95% CI 0.23–0.86); however, the risk estimation was based on nine cases. Conclusions: Our study shows a significantly increased EC risk in diabetic women as compared to the general population. In this study, a significantly higher EC risk was found in all patient groups by glucose-lowering medication combinations, except for insulin only users.
Customized Efficient Neural Network for COVID-19 Infected Region Identification in CT Images
Authors: Stefano Alessandro , Comelli Albert
Published: August 2021

Background: In the field of biomedical imaging, radiomics is a promising approach that aims to provide quantitative features from images. It is highly dependent on accurate identification and delineation of the volume of interest to avoid mistakes in the implementation of the texture-based prediction model. In this context, we present a customized deep learning approach aimed at addressing the real-time, and fully automated identification and segmentation of COVID-19 infected regions in computed tomography images. Methods: In a previous study, we adopted ENET, originally used for image segmentation tasks in self-driving cars, for whole parenchyma segmentation in patients with idiopathic pulmonary fibrosis which has several similarities to COVID-19 disease. To automatically identify and segment COVID-19 infected areas, a customized ENET, namely C-ENET, was implemented and its performance compared to the original ENET and some state-of-the-art deep learning architectures. Results: The experimental results demonstrate the effectiveness of our approach. Considering the performance obtained in terms of similarity of the result of the segmentation to the gold standard (dice similarity coefficient ~75%), our proposed methodology can be used for the identification and delineation of COVID-19 infected areas without any supervision of a radiologist, in order to obtain a volume of interest independent from the user. Conclusions: We demonstrated that the proposed customized deep learning model can be applied to rapidly identify, and segment COVID-19 infected regions to subsequently extract useful information for assessing disease severity through radiomics analyses.
Imaging with Coherent X-rays: From the Early Synchrotron Tests to SYNAPSE
Authors: Margaritondo Giorgio , Hwu Yeukuang
Published: August 2021

The high longitudinal and lateral coherence of synchrotron X-rays sources radically transformed radiography. Before them, the image contrast was almost only based on absorption. Coherent synchrotron sources transformed radiography into a multi-faceted tool that can extract information also from “phase” effects. Here, we report a very simple description of the new techniques, presenting them to potential new users without requiring a sophisticated background in advanced physics. We then illustrate the impact of such techniques with a number of examples. Finally, we present the international collaboration SYNAPSE (Synchrotrons for Neuroscience—an Asia-Pacific Strategic Enterprise), which targets the use of phase-contrast radiography to map one full human brain in a few years.
The Plant Salicylic Acid Signalling Pathway Regulates the Infection of a Biotrophic Pathogen in Grasses Associated with an Epichloë Endophyte
Authors: Kou Ming-Zhu , Bastías Daniel A. , Christensen Michael J. , Zhong Rui , Nan Zhi-Biao , Zhang Xing-Xu
Published: August 2021

The study of the contribution of the plant defence hormones, salicylic acid (SA) and jasmonic acid (JA), in the resistance against pathogens of plants associated with Epichloë fungal endophytes has been scanty. We hypothesised that Epichloë spp., capable of inducing host plant SA-dependent defences, would increase the levels of plant resistance against biotrophic pathogens. Plants of Achnatherum inebrians, with and without the fungal endophyte Epichloë gansuensis, were inoculated with the biotrophic fungal pathogen Blumeria graminis. We measured the status of plant defences (associated with SA and JA signalling pathways) and the levels of resistance to the pathogen. Plants associated with the endophyte showed less disease symptoms caused by the biotrophic pathogen than plants without the endophyte. In agreement with our hypothesis, the Epichloë endophyte increased the plant production of SA and enhanced the expression levels of plant genes of synthesis and response to the SA hormone. The elevated expression of SA-related genes coding for putative plant enzymes with anti-fungal activities promoted by the endophyte may explain the enhanced resistance to the pathogen. The present study highlights that interaction between the plant immune system and Epichloë fungal endophytes can contribute significantly to the resistance of endophyte-symbiotic plants against pathogens.