bluuebunny/crossref_metadata_2025 huggingface dataset:
Effective RNAi-Mediated Silencing of the Mismatch Repair MSH2 Gene Induces Sterility of Tomato Plants but Not an Increase in Meiotic Recombination
Authors: Strelnikova Svetlana R. , Krinitsina Anastasiya A. , Komakhin Roman A.
Published: July 2021

In plant breeding, the ability to manipulate meiotic recombination aids in the efficient construction of new allelic compositions of chromosomes and facilitates gene transfer from wild relatives of crop plants. The DNA mismatch repair system antagonizes meiotic recombination. In this research, a trial was conducted to evaluate transgenic tomato plants carrying an RNA interference (RNAi) construct designed to inhibit the expression of the mismatch repair MSH2 gene. To drive the RNAi construct, we used either a pro-SmAMP2 promoter from Stellaria media ANTIMICROBIAL PEPTIDE2 or a Cauliflower mosaic virus 35S promoter (CaMV35S). The results of real-time PCR showed that, with a 16 h light/8 h dark photoperiod, MSH2-RNAi tomato transgenic plants exhibited MSH2 gene transcript contents ranging from 0% to 3% in the leaves, relative to untransformed controls. However, with this lighting mode, the MSH2-RNAi transgenic plants grew slowly, flowered poorly, and did not form seed sets. During cultivation with a 12 h light/12 h dark photoperiod, MSH2-RNAi transgenic plants exhibited MSH2 gene transcript contents ranging from 3% to 42%, relative to untransformed controls. Under these conditions, F1 hybrid seed sets formed for most of the MSH2-RNAi transgenic plants with the RNAi construct driven by the CaMV35S promoter, and for one transformant with the RNAi construct driven by the pro-SmAMP2 promoter. Under conditions of a 12 h light/12 h dark photoperiod, most of the F1 transgenic hybrids showed MSH2 gene transcript contents ranging from 3% to 34% and formed F2 offspring sets, which made it possible to assess the meiotic recombination frequency. We showed that the effective inhibition of MSH2 in MSH2-RNAi tomato transgenic plants is not associated with an increase in meiotic recombination compared to the control, but it stimulates the sterility of plants. It was established that the expression of the MSH2 gene in tomato plants is about 50 times higher with a 12 h light/12 h dark than with a 16 h light/8 h dark photoperiod. It is discussed that, in Solanum lycopersicum tomato plants, which are not sensitive to the day length for flowering, changing the lighting time may be a means of controlling the meiotic recombination frequency within certain limits.
Extracellular Vesicle-Derived DNA vs. CfDNA as a Biomarker for the Detection of Colon Cancer
Authors: Thakur Kavita , Singh Manu Smriti , Feldstein-Davydova Sara , Hannes Victoria , Hershkovitz Dov , Tsuriel Shlomo
Published: July 2021

Liquid biopsy has emerged as a promising non-invasive way to diagnose tumor and monitor its progression. Different types of liquid biopsies have different advantages and limitations. In the present research, we compared the use of two types of liquid biopsy, extracellular vesicle-derived DNA (EV-DNA) and cell-free DNA (cfDNA) for identifying tumor mutations in patients with colon carcinoma. Method: DNA was extracted from the tumor tissue of 33 patients diagnosed with colon carcinoma. Targeted NGS panel, based on the hotspots panel, was used to identify tumor mutations. Pre-surgery serum and plasma were taken from the patients in which mutation was found in the tumor tissue. Extracellular vesicles were isolated from the serum followed by the extraction of EV-DNA. CfDNA was extracted from the plasma. The mutations found in the tumor were used to detect the circulating tumor DNA using ultra-deep sequencing. We compared the sensitivity of mutation detection and allele frequency obtained in EV-DNA and cfDNA. Results: The sensitivity of mutation detection in EV-DNA and cfDNA was 61.90% and 66.67%, respectively. We obtained almost identical sensitivity of mutation detection in EV-DNA and cfDNA in each of the four stages of colon carcinoma. The total DNA concentration and number mutant copies were higher in cfDNA vs. EV-DNA (p value = 0.002 and 0.003, respectively). Conclusion: Both cfDNA and EV-DNA can serve as tumor biomarkers. The use of EV-DNA did not lead to improved sensitivity or better detection of tumor DNA in the circulation.
Effectiveness of Elexacaftor/Tezacaftor/Ivacaftor Therapy in Three Subjects with the Cystic Fibrosis Genotype Phe508del/Unknown and Advanced Lung Disease
Authors: Terlizzi Vito , Colangelo Carmela , Marsicovetere Giovanni , D’Andria Michele , Francalanci Michela , Innocenti Diletta , Masi Eleonora , Avarello Angelo , Taccetti Giovanni , Amato Felice , Comegna Marika , Castaldo Giuseppe , Salvatore Donatello
Published: July 2021

We evaluated the effectiveness and safety of elexacaftor/tezacaftor/ivacaftor (ELX/TEZ/IVA) in three subjects carrying the Phe508del/unknown CFTR genotype. An ex vivo analysis on nasal epithelial cells (NEC) indicated a significant improvement of CFTR gating activity after the treatment. Three patients were enrolled in an ELX/TEZ/IVA managed-access program, including subjects with the highest percent predicted Forced Expiratory Volume in the 1st second (ppFEV1) < 40 in the preceding 3 months. Data were collected at baseline and after 8, 12 and 24 weeks of follow-up during treatment. All patients showed a considerable decrease of sweat chloride (i.e., meanly about 60 mmol/L as compared to baseline), relevant improvement of ppFEV1 (i.e., >8) and six-minute walk test, and an increase in body mass index after the first 8 weeks of treatment. No pulmonary exacerbations occurred during the 24 weeks of treatment and all domains of the CF Questionnaire-Revised improved. No safety concerns related to the treatment occurred. This study demonstrates the benefit from the ELX/TEZ/IVA treatment in patients with CF with the Phe508del and one unidentified CFTR variant. The preliminary ex vivo analysis of the drug response on NEC helps to predict the in vivo therapeutic endpoints.
Factors Influencing Physicians’ Continuous Blogging: A Survey
Authors: Chang I-Chiu , Cheng Kuei-Chen , Hsu Hui-Mei , Yen David C.
Published: July 2021

Background: Health information can be more easily transmitted and diffused through the Internet, but questionable online health information often misinforms patients. Physicians have a duty to inform patients how to achieve positive health outcomes. Many physicians often write blogs to provide patients with the right health information. However, most articles available on this subject only describe the blog phenomena without providing a theoretical background and an empirical analysis of doctors using blogs. Methods: This study based on social cognitive theory (SCT) explores the factors influencing physicians’ intention of continuously blogging. A total of 887 physician bloggers were invited to participate in an online survey and 128 valid responses were received. Results: The SCT was proven to be useful in explaining 36.8% of the variation in physicians’ continuous intention to blog. Conclusions: We provide references for platform developers with different strategies to motivate doctors to blog, and the implications and limitations of this study are discussed.
The Evidentiary Value of Late Nineteenth and Early Twentieth Century Postcards for Heritage Studies
Authors: Spennemann Dirk H. R.
Published: July 2021

A postcard ‘craze’ engulfed the developed world and colonial world during the end of the nineteenth and the beginning of the twentieth century, when over 2.3 million picture postcards were mailed in 1904 alone. Formally published picture postcards can provide a rich source of information for heritage studies as they depict landscape scenery, towns, individual buildings and public plantings such as parks. The evidentiary value of late nineteenth and early twentieth century postcards depends on the veracity of the depicted image. While based on photographs, processes of postcard production allowed the publisher to modify the original imagery to improve the messaging entailed in the image. Modes of image manipulation, such as retouching, can sufficiently alter the content of the image to create limitations to using published postcard imagery as a tool for historic landscape and building analysis. This is the first paper to systematically discuss the process of postcard production and the manipulation of images depicted on the view size of picture postcards. It demonstrates that where evidentiary emphasis is placed on postcard images, it is imperative that a systematic search for variants is carried out.
Design and Testing of a Pneumatic Oscillating Chinese Wolfberry Harvester
Authors: Chen Yun , Zhao Jian , Hu Guangrui , Chen Jun
Published: July 2021

A pneumatic oscillating Chinese wolfberry harvester was designed to improve the efficiency of manual harvesting and minimize fruit damage by mechanical harvesting. The shedding model of Chinese wolfberry under pneumatic forces was established based on kinetic analysis, and the flow field characteristics were investigated using computational fluid dynamics (CFD). The position and shape of the fan outlet were determined by comparing the effects of a linear reduction outlet and a Laval reduction outlet on the outlet airflow rate. The size of the oscillating mechanism was optimized with nonlinear constraints to achieve uniform oscillation of the airflow. A three-factor, three-level orthogonal test was conducted on the NingQi 7 wolfberry variety. The flow rate at the outlet, the oscillation frequency, and the outlet distance were the test factors, and the net harvesting rate of ripe fruits, the harvesting rate of green fruits, and the damage rate of ripe fruits were the test indices. The results showed that the net harvesting rate of ripe fruits was 85.21% at an outlet distance of 10 cm, an outlet flow rate of 70 m·s−1, and an oscillation frequency of 2 Hz. This study provides references for the design of pneumatic harvesters for Chinese wolfberry.
Effectiveness of the Functional and Cognitive Occupational Therapy (FaCoT) Intervention for Improving Daily Functioning and Participation of Individuals with Mild Stroke: A Randomized Controlled Trial
Authors: Adamit Tal , Shames Jeffrey , Rand Debbie
Published: July 2021

Background: Mild stroke can cause subtle cognitive–behavioral symptoms, which although might be hidden, can restrict community reintegration and participation. Cognitive rehabilitation programs exist for stroke but not specifically for mild stroke and the research evidence varies. The Functional and Cognitive Occupational Therapy (FaCoT) intervention was developed specifically for this population. Objective: To examine the effectiveness of FaCoT intervention for improving daily functioning and participation compared with standard care. Method: A single blind randomized controlled trial with assessments pre (T1), post (T2) and 3-month follow-up (T3). Individuals in the FaCoT group received 10 weekly sessions practicing cognitive and behavioral strategies. The Canadian Occupational Performance Measure (COPM) was the primary outcome measure, IADL-questionnaire, Reintegration to Normal Living questionnaire (RNL) were secondary measures. Results: In total, 66 community-dwelling individuals with mild stroke were randomly allocated to FaCoT (n = 33, mean (SD) age 64.6 (8.2), 33% women), or control group (n = 33, mean (SD) age 64.4 (10.8), 45% women). Time X Group interaction effects were found for the COPM performance (F(1.4,90.3) = 11.75, p < 0.000) and satisfaction (F(1.5,96.8) = 15.70, p < 0.000), with large effect size values. Significant between-group effects were found for RNL (F = 10.02, p < 0.002, ɳP2 = 0.13). Most participants in FaCoT achieved a clinically important difference in COPM between T1–T2, T1–T3, and in RNL between T1 to T3 compared with the control group. Conclusions: FaCoT intervention is effective to improve daily functioning, participation and satisfaction of individuals with mild stroke compared with standard care, therefore FaCoT should be implemented in community rehabilitation settings.
Descriptive Elements and Conceptual Structure of Glass Ceiling Research
Authors: Martínez-Fierro Salustiano , Lechuga Sancho María Paula
Published: July 2021

Women make up more than half of the population of every society and are seen as the creators and instructors of the next generation. They are potentially significant human resources in the scientific, social, and cultural development of most countries and, consequently, special attention needs to be paid to the form of their occupational promotion. This paper describes the evolution of research on glass ceiling until July 2020. We compiled a database of 823 articles focused on the field and published in academic journals in the ISI WoS database. Bibliometric methods and techniques were used to describe the evolution of scientific activity, countries, and active institutions, most productive authors, most relevant sources, most influential documents, trend topics, and social structure researched. This determined the state of the art and described the evolution of the literature in this field, and it will help scholars refine existing and initiate new research agendas. A total of 846 documents were identified, and the results showed an upward trend in glass ceiling scientific production. Based on these analyses, possible forms of future research are proposed to advance toward the consolidation of this scientific discipline.
Relationships between Depression, Daily Physical Activity, Physical Fitness, and Daytime Sleepiness among Japanese University Students
Authors: Shimamoto Hideki , Suwa Masataka , Mizuno Koh
Published: July 2021

In Japan and other developed countries, the lifestyles of many—particularly the younger generation—have been disrupted in recent years. This disruption may manifest as a decrease in physical activity and deterioration in the quality and quantity of sleep. Depressive tendencies are also increasing among university students. This study examined the relationship between mental health, physical activity, physical fitness, and daytime sleepiness. Eighty-five undergraduate students participated in this study (52 men and 33 women, aged 18.9 (±1.4) years). Physical activity levels were measured using an accelerometer (Lifecorder, Kenz, Nagoya, Japan) for two weeks. To evaluate their level of physical fitness, maximal oxygen uptake (V˙O2max) was calculated by an indirect method using a cycle ergometer. Depressive tendencies and daytime sleepiness were evaluated using the Patient Health Questionnaire (PHQ-9). The PHQ-9 score was positively correlated with sleepiness (r = 0.35, p = 0.001) and total steps per day (r = 0.39, p < 0.001). Moreover, the PHQ-9 score was positively correlated with V˙O2max (r = 0.25, p = 0.019). The PHQ-9 score was higher in students with good exercise habits or part-time jobs. An important finding was the positive correlation between depression and variables related to physical activity levels. These results suggest that vigorous physical activity, such as exercise and part-time jobs, might be positively associated with depressive tendencies among university students.
A Feeling of Otherness: A Qualitative Research Synthesis Exploring the Lived Experiences of Stigma in Individuals with Inflammatory Bowel Disease
Authors: Muse Kate , Johnson Emma , David Annabel L.
Published: July 2021

Inflammatory bowel disease (IBD) consists of Crohn’s disease and ulcerative colitis, chronic conditions involving inflammation and ulceration of the gastrointestinal tract. Individuals with IBD may be susceptible to experiencing health-related stigma: experienced, perceived, or internalised social exclusion, rejection, blame, or devaluation resulting from negative social judgements based on the disease. This qualitative research synthesis draws together findings from 38 studies describing lived experiences to develop a unified interpretative account of the experience of stigma in IBD. Analysis developed two categories: ‘The IBD journey’ explores the dynamic ways in which having IBD impacted on individuals’ self-identity and ‘a need to be understood’ examines the tension between wanting to be understood whilst feeling their true experiences needed to be hidden from or were misjudged by the social sphere. The overarching concept ‘feeling of otherness’ highlights that, rather than a static, binary experience, individuals moved across a continuum ranging from the excluding experience of feeling stigmatised and othered, to the inclusive experience of integration. Individuals fluctuated along this continuum across different physical, social, and health contexts. Psychological adjustment to IBD, drawing on experience of adaptive coping, and reconnecting with valued others through illness disclosure strengthened stigma resistance during more challenging times.
Automatic COVID-19 Detection Using Exemplar Hybrid Deep Features with X-ray Images
Authors: Barua Prabal Datta , Muhammad Gowdh Nadia Fareeda , Rahmat Kartini , Ramli Norlisah , Ng Wei Lin , Chan Wai Yee , Kuluozturk Mutlu , Dogan Sengul , Baygin Mehmet , Yaman Orhan , Tuncer Turker , Wen Tao , Cheong Kang Hao , Acharya U. Rajendra
Published: July 2021

COVID-19 and pneumonia detection using medical images is a topic of immense interest in medical and healthcare research. Various advanced medical imaging and machine learning techniques have been presented to detect these respiratory disorders accurately. In this work, we have proposed a novel COVID-19 detection system using an exemplar and hybrid fused deep feature generator with X-ray images. The proposed Exemplar COVID-19FclNet9 comprises three basic steps: exemplar deep feature generation, iterative feature selection and classification. The novelty of this work is the feature extraction using three pre-trained convolutional neural networks (CNNs) in the presented feature extraction phase. The common aspects of these pre-trained CNNs are that they have three fully connected layers, and these networks are AlexNet, VGG16 and VGG19. The fully connected layer of these networks is used to generate deep features using an exemplar structure, and a nine-feature generation method is obtained. The loss values of these feature extractors are computed, and the best three extractors are selected. The features of the top three fully connected features are merged. An iterative selector is used to select the most informative features. The chosen features are classified using a support vector machine (SVM) classifier. The proposed COVID-19FclNet9 applied nine deep feature extraction methods by using three deep networks together. The most appropriate deep feature generation model selection and iterative feature selection have been employed to utilise their advantages together. By using these techniques, the image classification ability of the used three deep networks has been improved. The presented model is developed using four X-ray image corpora (DB1, DB2, DB3 and DB4) with two, three and four classes. The proposed Exemplar COVID-19FclNet9 achieved a classification accuracy of 97.60%, 89.96%, 98.84% and 99.64% using the SVM classifier with 10-fold cross-validation for four datasets, respectively. Our developed Exemplar COVID-19FclNet9 model has achieved high classification accuracy for all four databases and may be deployed for clinical application.
Effects of Visfatin on Intracellular Mechanics and Catabolism in Human Primary Chondrocytes through Glycogen Synthase Kinase 3β Inactivation
Authors: Chang Shun-Fu , Huang Kuo-Chin , Lee Kuan-Han , Chiang Yao-Chang , Lee Wei-Ru , Hsieh Rong-Ze , Su Yu-Ping , Wu Shun-Chi
Published: July 2021

Osteoarthritis (OA) is still a recalcitrant musculoskeletal disease on account of its complex biochemistry and mechanical stimulations. Apart from stimulation by external mechanical forces, the regulation of intracellular mechanics in chondrocytes has also been linked to OA development. Recently, visfatin has received significant attention because of the clinical finding of the positive correlation between its serum/synovial level and OA progression. However, the precise mechanism involved is still unclear. This study determined the effect of visfatin on intracellular mechanics and catabolism in human primary chondrocytes isolated from patients. The intracellular stiffness of chondrocytes was analyzed by the particle-tracking microrheology method. It was shown that visfatin damages the microtubule and microfilament networks to influence intracellular mechanics to decrease the intracellular elasticity and viscosity via glycogen synthase kinase 3β (GSK3β) inactivation induced by p38 signaling. Further, microtubule network destruction in human primary chondrocytes is predominantly responsible for the catabolic effect of visfatin on the cyclooxygenase 2 upregulation. The present study shows a more comprehensive interpretation of OA development induced by visfatin through biochemical and biophysical perspectives. Finally, the role of GSK3β inactivation, and subsequent regulation of intracellular mechanics, might be considered as theranostic targets for future drug development for OA.
Epigenetic Regulation of Breast Cancer Stem Cells Contributing to Carcinogenesis and Therapeutic Implications
Authors: Wu Hsing-Ju , Chu Pei-Yi
Published: July 2021

Globally, breast cancer has remained the most commonly diagnosed cancer and the leading cause of cancer death among women. Breast cancer is a highly heterogeneous and phenotypically diverse group of diseases, which require different selection of treatments. Breast cancer stem cells (BCSCs), a small subset of cancer cells with stem cell-like properties, play essential roles in breast cancer progression, recurrence, metastasis, chemoresistance and treatments. Epigenetics is defined as inheritable changes in gene expression without alteration in DNA sequence. Epigenetic regulation includes DNA methylation and demethylation, as well as histone modifications. Aberrant epigenetic regulation results in carcinogenesis. In this review, the mechanism of epigenetic regulation involved in carcinogenesis, therapeutic resistance and metastasis of BCSCs will be discussed, and finally, the therapies targeting these biomarkers will be presented.
Copresence of High-Risk Human Papillomaviruses and Epstein–Barr Virus in Colorectal Cancer: A Tissue Microarray and Molecular Study from Lebanon
Authors: Nagi Karim , Gupta Ishita , Jurdi Nawaf , Yasmeen Amber , Vranic Semir , Batist Gerald , Moustafa Ala-Eddin Al
Published: July 2021

Colorectal cancer (CRC) is the third most common cause of cancer-related deaths worldwide. Human papillomaviruses (HPVs) and Epstein–Barr virus (EBV) have been reported to be present in different types of human cancers, including CRCs, where they can play a key role in the onset and/or progression of these cancers. Thus, we herein explored the prevalence of high-risk HPVs and EBV in a cohort of 94 CRC tissue samples and 13 colorectal normal tissues from the Lebanese population using polymerase chain reaction, immunohistochemistry, and tissue microarray methodologies. We found that high-risk HPVs are present in 64%, while EBV is present in 29% of our CRC samples. Additionally, our data showed that high-risk HPV types (16, 18, 35, 58, 51, 45, 52, 31, and 33) are the most frequent in CRC in the Lebanese cohort, respectively. Our data point out that HPVs and EBV are copresent in 28% of the samples. Thus, this study clearly suggests that high-risk HPVs and EBV are present/copresent in CRCs, where they could play an important role in colorectal carcinogenesis. Nevertheless, further investigations using a larger cohort are needed to elucidate the possible cooperation between these oncoviruses in the development of CRC.
Core Histones Are Constituents of the Perinuclear Theca of Murid Spermatozoa: An Assessment of Their Synthesis and Assembly during Spermiogenesis and Function after Gametic Fusion
Authors: Hamilton Lauren E. , Lion Morgan , Aguila Luis , Suzuki João , Acteau Genevieve , Protopapas Nicole , Xu Wei , Sutovsky Peter , Baker Mark , Oko Richard
Published: July 2021

The perinuclear theca (PT) of the eutherian sperm head is a cytoskeletal-like structure that houses proteins involved in important cellular processes during spermiogenesis and fertilization. Building upon our novel discovery of non-nuclear histones in the bovine PT, we sought to investigate whether this PT localization was a conserved feature of eutherian sperm. Employing cell fractionation, immunodetection, mass spectrometry, qPCR, and intracytoplasmic sperm injections (ICSI), we examined the localization, developmental origin, and functional potential of histones from the murid PT. Immunodetection localized histones to the post-acrosomal sheath (PAS) and the perforatorium (PERF) of the PT but showed an absence in the sperm nucleus. MS/MS analysis of selectively extracted PT histones indicated that predominately core histones (i.e., H3, H3.3, H2B, H2A, H2AX, and H4) populate the murid PT. These core histones appear to be de novo-synthesized in round spermatids and assembled via the manchette during spermatid elongation. Mouse ICSI results suggest that early embryonic development is delayed in the absence of PT-derived core histones. Here, we provide evidence that core histones are de novo-synthesized prior to PT assembly and deposited in PT sub-compartments for subsequent involvement in chromatin remodeling of the male pronucleus post-fertilization.
HERON: A Novel Tool Enables Identification of Long, Weakly Enriched Genomic Domains in ChIP-seq Data
Authors: Macioszek Anna , Wilczynski Bartek
Published: July 2021

The explosive development of next-generation sequencing-based technologies has allowed us to take an unprecedented look at many molecular signatures of the non-coding genome. In particular, the ChIP-seq (Chromatin ImmunoPrecipitation followed by sequencing) technique is now very commonly used to assess the proteins associated with different non-coding DNA regions genome-wide. While the analysis of such data related to transcription factor binding is relatively straightforward, many modified histone variants, such as H3K27me3, are very important for the process of gene regulation but are very difficult to interpret. We propose a novel method, called HERON (HiddEn MaRkov mOdel based peak calliNg), for genome-wide data analysis that is able to detect DNA regions enriched for a certain feature, even in difficult settings of weakly enriched long DNA domains. We demonstrate the performance of our method both on simulated and experimental data.
Relative Contribution of Different Mitochondrial Oxidative Phosphorylation Components to the Retinal Pigment Epithelium Barrier Function: Implications for RPE-Related Retinal Diseases
Authors: Guerra Michael H. , Yumnamcha Thangal , Singh Lalit P. , Ibrahim Ahmed S.
Published: July 2021

Disruption of retinal pigment epithelial (RPE) barrier integrity is involved in the pathology of several blinding retinal diseases including age-related macular degeneration (AMD) and diabetic retinopathy (DR), but the underlying causes and pathophysiology are not completely well-defined. Mitochondria dysfunction has often been considered as a potential candidate implicated in such a process. In this study, we aimed to dissect the role of different mitochondrial components; specifically, those of oxidative phosphorylation (OxPhos), in maintaining the barrier functionality of RPE. Electric cell-substrate impedance sensing (ECIS) technology was used to collect multi-frequency electrical impedance data to assess in real-time the barrier formation of the RPE cells. For this purpose, the human retinal pigment epithelial cell line—ARPE-19—was used and treated with varying concentrations of specific mitochondrial inhibitors that target different steps in OxPhos: Rotenone for complex I (the largest protein complex in the electron transport chain (ETC)); oligomycin for ATP synthase; and carbonyl cyanide-p-trifluoromethoxyphenyl hydrazone (FCCP) for uncoupling ATP synthesis from the accompanying ETC. Furthermore, data were modeled using the ECIS-Zθ software to investigate in depth the effects of these inhibitors on three separate barrier parameters: cell–cell interactions (Rb), cell–matrix interactions (α), and the cell membrane capacitance (Cm). The viability of ARPE-19 cells was determined by lactate dehydrogenase (LDH) Cytotoxicity Assay. The ECIS program’s modeling demonstrated that FCCP and thus OxPhos uncoupling disrupt the barrier function in the ARPE-19 cells across all three components of the total resistance (Rb, α, and Cm) in a dose-dependent manner. On the other hand, oligomycin and thus ATP synthase inhibition mostly affects the ARPE-19 cells’ attachment to their substrate evident by a significant decrease in α resistance in a dose-dependent manner, both at the end and throughout the duration of the experiment. On the contrary, rotenone and complex I inhibition mostly affect the ARPE-19 paracellular resistance Rb in a dose-dependent manner compared to basolateral resistance α or Cm. Our results clearly demonstrate differential roles for different mitochondrial components in maintaining RPE cell functionality in which uncoupling of OxPhos is a major contributing factor to the disruption barrier function. Such differences can be used in investigating gene expression as well as for screening of selective agents that improve the OxPhos coupling efficiency to be used in the therapeutic approach for treating RPE-related retinal diseases.
The Effect of Ethanol Treatment on the Quality of a New Table Grape Cultivar It 681–30 Stored at Low Temperature and after a 7-Day Shelf-Life Period at 20 °C: A Molecular Approach
Authors: Romero Irene , Vazquez-Hernandez Maria , Tornel Manuel , Escribano M. Isabel , Merodio Carmen , Sanchez-Ballesta M. Teresa
Published: July 2021

Despite the fact that many studies have examined the effectiveness of different gaseous postharvest treatments applied at low temperature to maintain table grape quality, the use of ethanol vapor has hardly been investigated. Thus, this work has studied the effectiveness of ethanol vapor-generating sachets in the maintenance of It 681–30 table grape quality, a new cultivar, during storage at low temperature and after the shelf-life period at 20 °C. To this end, various quality assessments have been carried out and the effect of the ethanol treatment on the expression of different genes (phenylpropanoids, transcription factors, PRs, and aquaporins) was determined. The results indicated that the application of ethanol vapor reduced the total decay incidence, weight loss, and the rachis browning index in It 681–30 grapes stored at 0 °C and after the shelf-life period at 20 °C, as compared to non-treated samples. Moreover, the modulation of STS7 and the different PR genes analyzed seems to play a part in the molecular mechanisms activated to cope with fungal attacks during the postharvest of It 681–30 grapes, and particularly during the shelf-life period at 20 °C. Furthermore, the expression of aquaporin transcripts was activated in samples showing higher weight loss. Although further work is needed to elucidate the role of ethanol in table grape quality, the results obtained in this work provide new insight into the transcriptional regulation triggered by ethanol treatment.
Monitoring Human Milk β-Casein Phosphorylation and O-Glycosylation Over Lactation Reveals Distinct Differences between the Proteome and Endogenous Peptidome
Authors: Dingess Kelly A. , Gazi Inge , van den Toorn Henk W. P. , Mank Marko , Stahl Bernd , Reiding Karli R. , Heck Albert J. R.
Published: July 2021

Human milk is a vital biofluid containing a myriad of molecular components to ensure an infant’s best start at a healthy life. One key component of human milk is β-casein, a protein which is not only a structural constituent of casein micelles but also a source of bioactive, often antimicrobial, peptides contributing to milk’s endogenous peptidome. Importantly, post-translational modifications (PTMs) like phosphorylation and glycosylation typically affect the function of proteins and peptides; however, here our understanding of β-casein is critically limited. To uncover the scope of proteoforms and endogenous peptidoforms we utilized mass spectrometry (LC-MS/MS) to achieve in-depth longitudinal profiling of β-casein from human milk, studying two donors across 16 weeks of lactation. We not only observed changes in β-casein’s known protein and endogenous peptide phosphorylation, but also in previously unexplored O-glycosylation. This newly discovered PTM of β-casein may be important as it resides on known β-casein-derived antimicrobial peptide sequences.
mTOR Activity and Autophagy in Senescent Cells, a Complex Partnership
Authors: Cayo Angel , Segovia Raúl , Venturini Whitney , Moore-Carrasco Rodrigo , Valenzuela Claudio , Brown Nelson
Published: July 2021

Cellular senescence is a form of proliferative arrest triggered in response to a wide variety of stimuli and characterized by unique changes in cell morphology and function. Although unable to divide, senescent cells remain metabolically active and acquire the ability to produce and secrete bioactive molecules, some of which have recognized pro-inflammatory and/or pro-tumorigenic actions. As expected, this “senescence-associated secretory phenotype (SASP)” accounts for most of the non-cell-autonomous effects of senescent cells, which can be beneficial or detrimental for tissue homeostasis, depending on the context. It is now evident that many features linked to cellular senescence, including the SASP, reflect complex changes in the activities of mTOR and other metabolic pathways. Indeed, the available evidence indicates that mTOR-dependent signaling is required for the maintenance or implementation of different aspects of cellular senescence. Thus, depending on the cell type and biological context, inhibiting mTOR in cells undergoing senescence can reverse senescence, induce quiescence or cell death, or exacerbate some features of senescent cells while inhibiting others. Interestingly, autophagy—a highly regulated catabolic process—is also commonly upregulated in senescent cells. As mTOR activation leads to repression of autophagy in non-senescent cells (mTOR as an upstream regulator of autophagy), the upregulation of autophagy observed in senescent cells must take place in an mTOR-independent manner. Notably, there is evidence that autophagy provides free amino acids that feed the mTOR complex 1 (mTORC1), which in turn is required to initiate the synthesis of SASP components. Therefore, mTOR activation can follow the induction of autophagy in senescent cells (mTOR as a downstream effector of autophagy). These functional connections suggest the existence of autophagy regulatory pathways in senescent cells that differ from those activated in non-senescence contexts. We envision that untangling these functional connections will be key for the generation of combinatorial anti-cancer therapies involving pro-senescence drugs, mTOR inhibitors, and/or autophagy inhibitors.
Microbiota Signals during the Neonatal Period Forge Life-Long Immune Responses
Authors: Phillips-Farfán Bryan , Gómez-Chávez Fernando , Medina-Torres Edgar Alejandro , Vargas-Villavicencio José Antonio , Carvajal-Aguilera Karla , Camacho Luz
Published: July 2021

The microbiota regulates immunological development during early human life, with long-term effects on health and disease. Microbial products include short-chain fatty acids (SCFAs), formyl peptides (FPs), polysaccharide A (PSA), polyamines (PAs), sphingolipids (SLPs) and aryl hydrocarbon receptor (AhR) ligands. Anti-inflammatory SCFAs are produced by Actinobacteria, Bacteroidetes, Firmicutes, Spirochaetes and Verrucomicrobia by undigested-carbohydrate fermentation. Thus, fiber amount and type determine their occurrence. FPs bind receptors from the pattern recognition family, those from commensal bacteria induce a different response than those from pathogens. PSA is a capsular polysaccharide from B. fragilis stimulating immunoregulatory protein expression, promoting IL-2, STAT1 and STAT4 gene expression, affecting cytokine production and response modulation. PAs interact with neonatal immunity, contribute to gut maturation, modulate the gut–brain axis and regulate host immunity. SLPs are composed of a sphingoid attached to a fatty acid. Prokaryotic SLPs are mostly found in anaerobes. SLPs are involved in proliferation, apoptosis and immune regulation as signaling molecules. The AhR is a transcription factor regulating development, reproduction and metabolism. AhR binds many ligands due to its promiscuous binding site. It participates in immune tolerance, involving lymphocytes and antigen-presenting cells during early development in exposed humans.
Growth Performance, Waste Reduction Efficiency and Nutritional Composition of Black Soldier Fly (Hermetia illucens) Larvae and Prepupae Reared on Coconut Endosperm and Soybean Curd Residue with or without Supplementation
Authors: Pliantiangtam Nichaphon , Chundang Pipatpong , Kovitvadhi Attawit
Published: July 2021

Black soldier fly (BSF, Hermetia illucens) larvae are considered as insects with a high potential to convert organic waste into high-value products. The objective of this study was to investigate the growth performance, waste reduction efficiency, and nutritional composition of BSF reared on different ratios of coconut endosperm (C) and soybean curd residue (S), with or without supplementation, compared to standard diets (Gainesville: G and starter chicken diet: CK). Seven-day-old larvae were randomly divided into eight experimental groups (G, CK, and three different ratios of C and S with or without supplementation) with three replicates with an equal weight of larvae. The supplement contained calcium, phosphorus, amino acids, and a mineral–vitamin premix which was formulated to correlate with CK. Each replicate was terminated, measured, and evaluated when 40% of larvae had reached prepupal stage. The highest larval weight gain was presented in BSF fed CK, followed by those fed coconut endosperm and soybean curd residue at a ratio of 20:80 (C20S80), and coconut endosperm and soybean curd residue at a ratio of 50:50 (C50S50) without supplementation (numbers after C and S represent their percentage in the formulation; p < 0.001). Harvesting was delayed in the BSF fed C80S20 with and without supplementation (p < 0.001). The number of total larvae and prepupae was not significantly different between groups (p > 0.05). The greatest waste reduction efficiency was observed in the G, C50S50, and C20S80 groups without supplementation (p < 0.001). All groups with supplementation had a higher proportion of ash in both larvae and prepupae compared to non-supplemented groups (p < 0.001), but lower growth performance. The highest percentage of crude protein in larvae was presented in the Gainesville and C20S80 groups followed by the CK and C50S50 groups (p < 0.001). Equal proportions of C and S without supplementation are suggested as a rearing substrate. However, growth performance was lower than for CK; therefore, further studies could investigate cost-efficient techniques to promote this parameter.
Abundant Citizen Science Data Reveal That the Peacock Butterfly Aglais io Recently Became Bivoltine in Belgium
Authors: Herremans Marc , Gielen Karin , Van Kerckhoven Jos , Vanormelingen Pieter , Veraghtert Wim , Swinnen Kristijn R.R. , Maes Dirk
Published: July 2021

The peacock butterfly is abundant and widespread in Europe. It is generally believed to be univoltine (one generation per year): adults born in summer overwinter and reappear again in spring to reproduce. However, recent flight patterns in western Europe mostly show three peaks during the year: a first one in spring (overwintering butterflies), a second one in early summer (offspring of the spring generation), and a third one in autumn. It was thus far unclear whether this autumn flight peak was a second new generation or consisted of butterflies flying again in autumn after a summer rest (aestivation). The life cycle of one of Europe’s most common butterflies is therefore still surprisingly inadequately understood. We used hundreds of thousands of observations and thousands of pictures submitted by naturalists from the public to the online portal observation.orgin Belgium and analyzed relations between flight patterns, condition (wear), reproductive cycles, peak abundances, and phenology to clarify the current life history. We demonstrate that peacocks have shifted towards two new generations per year in recent decades. Mass citizen science data in online portals has become increasingly important in tracking the response of biodiversity to rapid environmental changes such as climate change.
Contralateral C7 Nerve Transfer for Stroke Recovery: New Frontier for Peripheral Nerve Surgery
Authors: Alawieh Ali M. , Au Yong Nicholas , Boulis Nicholas M.
Published: July 2021

Ischemic stroke remains a major cause of disability in the United States and worldwide. Following the large-scale implementation of stroke thrombectomy and the optimization of treatment protocols for acute stroke, the reduction in stroke-associated mortality has resulted in an increased proportion of stroke survivors, many of whom have moderate to severe disability. To date, the treatment of subacute and chronic stroke has remained a challenge. Several approaches, involving pharmacological interventions to promote neuroplasticity, brain stimulation strategies and rehabilitative interventions, are currently being explored at different stages of the translational spectrum, yet level 1 evidence is still limited. In a recent landmark study, surgical intervention using contralateral C7 nerve transfer, an approach used to treat brachial plexus injury, was implemented in patients with chronic stroke, demonstrating an added benefit to standard rehabilitation strategies, leading to improved motor performance and reduced spasticity. The procedure involved the transfer of the C7 nerve root and middle trunk from the uninjured extremity to the injured extremity using a short conduit that allows for faster regeneration and innervation of the injured upper extremity via the ipsilateral (contralesional) hemisphere. In this work, we review the rationale for using contralateral C7 nerve transfer in stroke, describe the surgical intervention with associated variations and limitations, and discuss the current evidence for the efficacy of this technique in ischemic stroke research.
Automated Segmentation of the Human Abdominal Vascular System Using a Hybrid Approach Combining Expert System and Supervised Deep Learning
Authors: Lareyre Fabien , Adam Cédric , Carrier Marion , Raffort Juliette
Published: July 2021

Background: Computed tomography angiography (CTA) is one of the most commonly used imaging technique for the management of vascular diseases. Here, we aimed to develop a hybrid method combining a feature-based expert system with a supervised deep learning (DL) algorithm to enable a fully automatic segmentation of the abdominal vascular tree. Methods: We proposed an algorithm based on the hybridization of a data-driven convolutional neural network and a knowledge-based model dedicated to vascular system segmentation. By using two distinct datasets of CTA from patients to evaluate independence to training dataset, the accuracy of the hybrid method for lumen and thrombus segmentation was evaluated compared to the feature-based expert system alone and to the ground truth provided by a human expert. Results: The hybrid approach demonstrated a better accuracy for lumen segmentation compared to the expert system alone (volume similarity: 0.8128 vs. 0.7912, p = 0.0006 and Dice similarity coefficient: 0.8266 vs. 0.7942, p < 0.0001). The accuracy for thrombus segmentation was also enhanced using the hybrid approach (volume similarity: 0.9404 vs. 0.9185, p = 0.0027 and Dice similarity coefficient: 0.8918 vs. 0.8654, p < 0.0001). Conclusions: By enabling a robust and fully automatic segmentation, the method could be used to develop real-time decision support to help in the management of vascular diseases.
The Role of Dendritic Cells during Physiological and Pathological Dentinogenesis
Authors: Quispe-Salcedo Angela , Ohshima Hayato
Published: July 2021

The dental pulp is a soft connective tissue of ectomesenchymal origin that harbors distinct cell populations, capable of interacting with each other to maintain the vitality of the tooth. After tooth injuries, a sequence of complex biological events takes place in the pulpal tissue to restore its homeostasis. The pulpal response begins with establishing an inflammatory reaction that leads to the formation of a matrix of reactionary or reparative dentin, according to the nature of the exogenous stimuli. Using several in vivo designs, antigen-presenting cells, including macrophages and dendritic cells (DCs), are identified in the pulpal tissue before tertiary dentin deposition under the afflicted area. However, the precise nature of this phenomenon and its relationship to inherent pulp cells are not yet clarified. This literature review aims to discuss the role of pulpal DCs and their relationship to progenitor/stem cells, odontoblasts or odontoblast-like cells, and other immunocompetent cells during physiological and pathological dentinogenesis. The concept of “dentin-pulp immunology” is proposed for understanding the crosstalk among these cell types after tooth injuries, and the possibility of immune-based therapies is introduced to accelerate pulpal healing after exogenous stimuli.
Intestinal Microbial Metabolites in Ankylosing Spondylitis
Authors: Scalise Giuseppe , Ciancio Antonio , Mauro Daniele , Ciccia Francesco
Published: July 2021

Ankylosing spondylitis (AS) is a chronic inflammatory disease characterized by inflammation of axial joints and the pelvis. It is known that intestinal dysbiosis may exert direct pathogenic effects on gut homeostasis and may act as a triggering factor for the host innate immune system to activate and cause inflammation in extraintestinal sites in the so-called “gut-joint axis”, contributing to AS pathogenesis. However, although the intestinal microbiota’s influence on the clinical manifestation of AS is widely accepted, the mechanisms mediating the cross-talk between the intestinal lumen and the immune system are still not completely defined. Recent evidence suggests that the metabolism of microbial species may be a source of metabolites and small molecules participating in the complex network existing between bacteria and host cells. These findings may give inputs for further research of novel pharmacological targets and pave the way to applying dietary interventions to prevent the onset and ameliorate the clinical presentation of the disease. In this review, we discuss the role of some of the biological mediators of microbial origin, with a particular focus on short-chain fatty acids, tryptophan and vitamin B derivatives, and their role in barrier integrity and type 3 immunity in the context of AS.