EXCLI Journal https://www.excli.de/index.php/excli <center><img class="img-responsive" src="/public/site/images/lindemann/Lucida_logo_neu_geschrieben.PNG" alt="EXCLI Journal Logo"></center> <p>EXCLI Journal publishes original research reports, authoritative reviews and case reports of experimental and clinical sciences.</p> <div class="toggleBox"><input id="toggleContent" name="toggleContent" type="checkbox"> <label class="open" for="toggleContent">More...</label> <label class="close" for="toggleContent">Less...</label> <div> <p><strong>EXCLI Journal </strong>(eISSN 1611-2156)&nbsp;is particularly keen to keep a broad view of science and technology, and therefore welcomes papers which bridge disciplines and may not suit the narrow specialism of other journals. Although the general emphasis is on biological sciences, studies from the following fields are explicitly encouraged:</p> <p>Immunology, toxicology, ergonomics, neurosciences, psychology, occupational medicine, clinical and preclinical studies, drug development, pharmacology, environmental health, chemistry including analytical chemistry, biochemistry, cell biology, genetics, forensic medicine, oncology and cancer research, proteomics, systems biology, hepatology and gastroenterology, aging research, psychiatric research, behavioral sciences.</p> </div> </div> IfADo - Leibniz Research Centre for Working Environment and Human Factors, Dortmund en-US EXCLI Journal 1611-2156 <p>Authors who publish in this journal agree to the following terms:</p> <ul> <li>The authors keep the copyright and grant the journal the right of first publication under the terms of the Creative Commons Attribution license, <a href="https://creativecommons.org/licenses/by/4.0/" target="_blank" rel="noopener">CC BY 4.0</a>. This licencse permits unrestricted use, distribution and reproduction in any medium, provided that the original work is properly cited.</li> <li>The use of general descriptive names, trade names, trademarks, and so forth in this publication, even if not specifically identified, does not imply that these names are not protected by the relevant laws and regulations.</li> <li>Because the advice and information in this journal are believed to be true and accurate at the time of publication, neither the authors, the editors, nor the publisher accept any legal responsibility for any errors or omissions presented in the publication. The publisher makes no guarantee, express or implied, with respect to the material contained herein.</li> <li>The authors can enter into additional contracts for the non-exclusive distribution of the journal's published version by citing the initial publication in this journal (e.g. publishing in an institutional repository or in a book).</li> </ul> Comparative highlights on MERS-COV, SARS-COV-1, SARS-COV-2, and NEO-COV https://www.excli.de/index.php/excli/article/view/5355 <p>The Severe Acute Respiratory Syndrome (SARS-CoV), Middle East Respiratory Syndrome (MERS-CoV), Neo-CoV, and 2019 novel coronavirus (SARS-CoV-2/COVID-19), are the most notable coronaviruses, infecting the number of people worldwide by targeting the respiratory system. All these viruses are of zoonotic origin, predominantly from bats, which are one of the natural reservoir hosts for coronaviruses. Thus, the major goal of our review article is to compare and contrast the characteristics, and attributes of these coronaviruses. While SARS-CoV-1, MERS-CoV, and COVID-19 have many viral similarities due to their classification, they are not genetically related. COVID-19 shares approximately 79% of its genome with SARS-CoV-1, and about 50% with MERS-CoV. The shared receptor protein, ACE2, is one of the most striking genetic similarities between SARS-CoV-1 and SARS-CoV-2. The SARS-CoV primarily replicates in the epithelial cells of the respiratory system, but it may also affect macrophages, monocytes, activated T cells, and dendritic cells. On the other hand, MERS-CoV, not only infects the epithelial and immune cells, but it may replicate also and lyse them, which is one of the common reasons for MERS's higher mortality rate. The details of infections caused by SARS-CoV-2 and lytic replication mechanisms in host cells are currently mysterious. In this review article, we will discuss the comparative highlights of SARS-CoV-1, MERS-CoV, SARS-CoV-2, and Neo-CoV, concerning their structural features and morphological characteristics, sources of virus origin and their evolutionary transitions, infection mechanism, computational study approaches, pathogenesis and their severity towards several diseases, possible therapeutic approaches, and preventive measures.</p> Rajat Goyal Rupesh Gautam Hitesh Chopra Ankit Kumar Dubey Rajeev K. Singla Rehab A. Rayan Mohammad Amjad Kamal Copyright (c) 2022 Rajat Goyal, Rupesh Gautam, Hitesh Chopra, Ankit Kumar Dubey, Rajeev K. Singla, Rehab A. Rayan, Mohammad Amjad Kamal http://creativecommons.org/licenses/by/4.0 2022-09-29 2022-09-29 21 1245 1272 10.17179/excli2022-5355 COVID-19 and its genomic variants https://www.excli.de/index.php/excli/article/view/5315 <p>Coronavirus disease-19 (COVID-19), caused by a β-coronavirus and its genomic variants, is associated with substantial morbidities and mortalities globally. The COVID-19 virus and its genomic variants enter host cells upon binding to the angiotensin converting enzyme 2 receptors that are expressed in a variety of tissues, but predominantly in the lungs, heart, and blood vessels. Patients afflicted with COVID-19 may be asymptomatic or present with critical symptoms possibly due to diverse lifestyles, immune responses, aging, and underlying medical conditions. Geriatric populations, especially men in comparison to women, with immunocompromised conditions, are most vulnerable to severe COVID-19 associated infections, complications, and mortalities. Notably, whereas immunomodulation, involving nutritional consumption, is essential to protecting an individual from COVID-19, immunosuppression is detrimental to a person with this aggressive disease. As such, immune health is inversely correlated to COVID-19 severity and resulting consequences. Advances in genomic and proteomic technologies have helped us to understand the molecular events underlying symptomatology, transmission and, pathogenesis of COVID-19 and its genomic variants. Accordingly, there has been development of a variety of therapeutic interventions, ranging from mask wearing to vaccination to medication. This review summarizes the current understanding of molecular pathogenesis of COVID-19, effects of comorbidities on COVID-19, and prospective therapeutic strategies for the prevention and treatment of this contagious disease.</p> Pulak R. Manna Zackery C. Gray Malabika Sikdar P. Hemachandra Reddy Copyright (c) 2022 Pulak R. Manna, Zackery C. Gray, Malabika Sikdar, P. Hemachandra Reddy http://creativecommons.org/licenses/by/4.0 2022-09-13 2022-09-13 21 1196 1221 10.17179/exli2022-5315 Berberine https://www.excli.de/index.php/excli/article/view/5234 <p class="Abstract"><span lang="EN-US">Proprotein convertase subtilisin/kexin type 9 (PCSK9) is a secreted protein and a critical regulator of low-density lipoprotein-cholesterol (LDL-C) through inducing degradation of the LDL receptor (LDLR) within the hepatocyte lysosome. PCSK9 deficiency significantly improves the survival rate of cardiovascular disease (CVDs) patients. Up to now, various PCSK9 inhibition approaches have been tested. However, the currently available PCSK9 inhibitors' widespread use is limited due to their inconvenient method of administration and high cost. On the other hand, inhibiting PCSK9 with nutraceuticals is safe and affordable. The plant-derived compound berberine has shown anti-PCSK9 activity in several studies. Berberine is an isoquinoline quaternary alkaloid of phyto origin. Berberine treatment boosts the hepatic expression of LDLRs, while decreasing the expression and secretion of the LDLR modulator PCSK9. The current review presents a collection of <em>in vitro </em>and<em> in vivo</em> studies investigating berberine's effects on PCSK9 mRNA expression, protein level, and function.</span></p> Sarina Ataei Prashant Kesharwani Amirhossein Sahebkar Copyright (c) 2022 Sarina Ataei, Prashant Kesharwani, Amirhossein Sahebkar http://creativecommons.org/licenses/by/4.0 2022-08-17 2022-08-17 21 1099 1110 10.17179/excli2022-5234 Bimanual motor impairments in older adults https://www.excli.de/index.php/excli/article/view/5236 <p>This updated systematic review and meta-analysis further examined potential effects of aging on bimanual movements. Forty-seven qualified studies that compared bimanual motor performances between elderly and younger adults were included in this meta-analysis. Moderator variable analyses additionally determined whether altered bimanual motor performances in older adults were different based on the task types (i.e., symmetry vs. asymmetry vs. complex) or outcome measures (i.e., accuracy vs. variability vs. movement time). The random effects model meta-analysis on 80 comparisons from 47 included studies revealed significant negative overall effects indicating more bimanual movement impairments in the elderly adults than younger adults. Moderator variable analyses found that older adults showed more deficits in asymmetrical bimanual movement tasks than symmetrical and complex tasks, and the bimanual movement impairments in the elderly adults included less accurate, more variable, and greater movement execution time than younger adults. These findings suggest that rehabilitation programs for improving motor actions in older adults are necessary to focus on functional recovery of interlimb motor control including advanced motor performances as well coordination.</p> Nyeonju Kang Do Kyung Ko James H. Cauraugh Copyright (c) 2022 Nyeonju Kang, Do Kyung Ko, James H. Cauraugh http://creativecommons.org/licenses/by/4.0 2022-08-16 2022-08-16 21 1068 1083 10.17179/excli2022-5236 Nanocarriers for delivery of siRNA as gene silencing mediator https://www.excli.de/index.php/excli/article/view/4975 <p class="Abstract"><span lang="EN-US">The term nanocarrier refers to sub-micrometric particles of less than 100 nm, designed to transport, distribute, and release nanotechnology-based drug delivery systems. siRNA therapy is a novel strategy that has great utility for a variety of treatments, however naked siRNA delivery <span style="color: #222222;">has not been an effective strategy, resulting in the necessary use of nanocarriers for delivery. </span>This review aims to highlight the versatility of carriers based on smart drug delivery systems. The nanocarriers based on nanoparticles as siRNA DDS have provided a set of very attractive advantages related to improved physicochemical properties, such as high surface-to-volume ratio, versatility to package siRNA, provide a dual function to both protect extracellular barriers that lead to elimination and overcome intracellular barriers limiting cytosolic delivery, and possible chemical modifications on the nanoparticle surface to improve stability and targeting. <span style="color: #1c1d1e;">Lipid and polymeric nanocarriers have</span> proven to be stable, biocompatible, and effective <em>in vitro</em>,<span style="color: #1c1d1e;"> further exploration of the development of new nanocarriers is needed to obtain safe and biocompatible tools for effective therapy.</span></span></p> Aideé Morales-Becerril Liliana Aranda-Lara Keila Isaac-Olivé Blanca E. Ocampo-García Enrique Morales-Avila Copyright (c) 2022 Aideé Morales-Becerril, Liliana Aranda-Lara, Keila Isaac-Olivé, Blanca E. Ocampo-García, Enrique Morales-Avila http://creativecommons.org/licenses/by/4.0 2022-08-01 2022-08-01 21 1028 1052 10.17179/excli2022-4975 Pivotal role of vitamin D in mitochondrial health, cardiac function, and human reproduction https://www.excli.de/index.php/excli/article/view/4935 <p class="Abstract"><span lang="EN-US">Vitamin D, a secosteroid hormone, appears to have significant beneficial effects on various physiological systems, including the musculoskeletal system. Vitamin D assists in the regulation of numerous critical biological functions and physiological processes in humans, including inflammation, oxidative stress, and mitochondrial respiration, and is also linked to cardiac diseases. It is also reported that vitamin D plays a central role in molecular and cellular mechanisms, which reduce oxidative stress, and tissue damage and regulate cellular health. On the other side, hypovitaminosis D reduces mitochondrial activity and increases oxidative stress and inflammation in the body. Hypervitaminosis D increases the prevalence and severity of cellular damage. It has also been reported that vitamin D is involved in many functions of the reproductive system in human and critically play an important role in the reproductive tissues of women and men. Its role is very well defined, starting from female menarche to menopause, pregnancy, and lactation, and finally in male fertility. Hence, the appropriate amount of vitamin D is necessary to maintain the normal function of cell organelles. Based on recent studies, it is understood that vitamin D is involved in the biological activities of mitochondria in cells, especially in cardiomyocytes. In this review, we emphasized the role of vitamin D in mitochondrial respiration, which could significantly influence heart health and human reproduction. </span></p> Alavala Matta Reddy Mumtaz Iqbal Hitesh Chopra Shaheda Urmi Sunil Junapudi Shabana Bibi Santosh Kumar Gupta Viajaya Nirmala Pangi Inderbir Singh Mohamed M. Abdel-Daim Copyright (c) 2022 Alavala Matta Reddy, Mumtaz Iqbal, Hitesh Chopra, Shaheda Urmi, Sunil Junapudi, Shabana Bibi, Santosh Kumar Gupta, Viajaya Nirmala Pangi, Inderbir Singh, Mohamed M. Abdel-Daim http://creativecommons.org/licenses/by/4.0 2022-07-20 2022-07-20 21 967 990 10.17179/excli2022-4935 Selenium as an important factor in various disease states https://www.excli.de/index.php/excli/article/view/5137 <p class="Abstract"><span lang="EN-US">Selenium (Se) is an element that has a pro-health effect on humans and animals. However, both the deficiency of this element and its excess may prove harmful to the body depending on the chemical form of the selenium, the duration of supplementation, and the human health condition. Many data indicate insufficient coverage of the demand for selenium in humans and animals due to its low content in soils and food products. A balance in the physiological process of the body can be achieved via the proper percentage of organically active minerals in the feed of animals as well as human beings. Selenium is a trace mineral of great importance to the body, required for the maintenance of a variety of its processes; primarily, selenium maintains immune endocrine, metabolic, and cellular homeostasis. Recently, this element has been emerging as a most promising treatment option for various disorders. Therefore, research based on Se has been increasing in recent times. The present review is designed to provide up-to-date information related to Se and its different forms as well as its effects on health.</span></p> Marek Kieliszek Iqra Bano Copyright (c) 2022 Marek Kieliszek, Iqra Bano http://creativecommons.org/licenses/by/4.0 2022-07-05 2022-07-05 21 948 966 10.17179/excli2022-5137 Insights into the mechanisms of non-coding RNAs’ implication in the pathogenesis of Alzheimer’s disease https://www.excli.de/index.php/excli/article/view/5006 <p class="Abstract"><span lang="EN-US">Non-coding RNAs including long non-coding RNAs (lncRNAs) and microRNAs (miRNAs) are implicated in the regulation of gene expression at transcriptional, posttranscriptional, and epigenetic levels. Several studies in cell lines, animal models, and humans, have revealed that non-coding RNAs play crucial roles in the pathogenesis of Alzheimer's disease (AD). Detailed knowledge on their mechanism of implication in the AD pathogenesis can help to develop novel therapeutic and disease management strategies. The two main pathological hallmarks of AD are amyloid plaques resulting from the β-amyloid accumulation, and neurofibrillary tangles (NFT) due to the phosphorylated tau accumulation. Several lncRNAs and miRNAs play crucial roles in both these hallmarks of the AD pathogenesis and other AD-related pathological procedures such as neuronal and synaptic plasticity, neuroinflammation, neuronal differentiation and neuronal apoptosis. In this review, we outlined the non-coding RNAs and further discussed how they are implicated in these AD-related pathological procedures.</span></p> Majid Khodayi-Shahrak Mohammad Khalaj-Kondori Mohammad Ali Hosseinpour Feizi Mahnaz Talebi Copyright (c) 2022 Majid Khodayi-Shahrak, Mohammad Khalaj-Kondori, Mohammad Ali Hosseinpour Feizi, Mahnaz Talebi http://creativecommons.org/licenses/by/4.0 2022-06-30 2022-06-30 21 921 940 10.17179/excli2022-5006 Elevated risk of thrombotic manifestations of SARS-CoV-2 infection in cancer patients https://www.excli.de/index.php/excli/article/view/5073 <p>Coronavirus disease 2019 (COVID-19) results in higher risks of hospitalization or death in older patients and those with multiple comorbidities, including malignancies. Patients with cancer have greater risks of COVID-19 onset and worse prognosis. This excess is mainly explained by thrombotic complications. Indeed, an imbalance in the equilibrium between clot formation and bleeding, increased activation of coagulation, and endothelial dysfunction characterize both COVID-19 patients and those with cancer. With this review, we provide a summary of the pathological mechanisms of coagulation and thrombotic manifestations in these patients and discuss the possible therapeutic implications of these phenomena.</p> Annunziata Stefanile Maria Cellerino Tatiana Koudriavtseva Copyright (c) 2022 Annunziata Stefanile, Maria Cellerino, Tatiana Koudriavtseva http://creativecommons.org/licenses/by/4.0 2022-06-30 2022-06-30 21 906 920 10.17179/excli2022-5073 Effects of curcumin on low-density lipoprotein oxidation https://www.excli.de/index.php/excli/article/view/4878 <p class="Abstract"><span lang="EN-US">Atherosclerosis is the most frequent cause of death globally. Oxidized low-density lipoprotein (ox-LDL) has an essential role in the formation of atherosclerotic plaques and foamy macrophages. Ox-LDL increases the uptake of cholesterol by macrophages and is the major cause of blood flow disruption. Ox-LDL is produced during oxidative stress and treatment with antioxidants could inhibit the production and function of ox-LDL. Curcumin is a potent antioxidant and has a strong track record in the treatment of numerous diseases. Recent studies indicate that Curcumin exerts a lipid-lowering effect, and can modulate the formation of atherosclerotic plaque. The current review focuses upon the role of Curcumin in oxidation of LDL and foam cell formation in atherosclerotic lesions. </span></p> Fatemeh Baratzadeh Alexandra E. Butler Prashant Kesharwani Seyed Adel Moallem Amirhossein Sahebkar Copyright (c) 2022 Fatemeh Baratzadeh, Alexandra E. Butler, Prashant Kesharwani, Seyed Adel Moallem, Amirhossein Sahebkar http://creativecommons.org/licenses/by/4.0 2022-06-22 2022-06-22 21 840 851 10.17179/excli2022-4878 The association of ACE1, ACE2, TMPRSS2, IFITM3 and VDR polymorphisms with COVID-19 severity https://www.excli.de/index.php/excli/article/view/4976 <p class="Abstract"><span lang="EN-US">Genes involved in the regulation of viral recognition and its entry into a host cell have been identified as candidates for genetic association studies on COVID-19 severity. Published findings on the effects of polymorphisms within <em>ACE1</em>, <em>ACE2</em>, <em>TMPRSS2</em>, <em>IFITM3</em> and <em>VDR</em> genes remained inconclusive, so we conducted a systematic review and meta-analysis in order to elucidate their potential involvement in the genetic basis underlying the severity of COVID-19 and/or an outcome of SARS-CoV-2 infection. Identification of potentially eligible studies was based on PubMed, Scopus and Web of Science database search. Relevant studies (n=29) with a total number of 8247 SARS-CoV-2-positive participants were included in qualitative synthesis, while results of 21 studies involving 5939 were pooled in meta-analysis. Minor allele I of rs1799752 located within <em>ACE1</em> was identified as a protective variant against severe COVID-19, while its effect on mortality rate was opposite. Similarly, minor allele A of <em>ACE2</em> polymorphism, rs2285666, was found to associate with a decreased risk of severe COVID-19 (<em>P</em> = 0.003, OR = 0.512, 95 % CI = 0.331-0.793). Statistical significance was also seen for the association between COVID-19 severity and rs12329760 located within <em>TMPRSS2</em>. Our results did not support the supposed association of rs12252 in <em>IFITM3</em> and polymorphisms within <em>VDR</em> with disease severity. We conclude that genetic variants within <em>ACE1</em>, <em>ACE2</em> and <em>TMPRSS2</em> may be potential biomarkers of COVID-19 severity, which needs to be further confirmed in a larger set of studies.</span></p> Zorana Dobrijević Dragana Robajac Nikola Gligorijević Miloš Šunderić Ana Penezić Goran Miljuš Olgica Nedić Copyright (c) 2022 Zorana Dobrijević, Dragana Robajac, Nikola Gligorijević, Miloš Šunderić, Ana Penezić, Goran Miljuš, Olgica Nedić http://creativecommons.org/licenses/by/4.0 2022-06-20 2022-06-20 21 818 839 10.17179/excli2022-4976 Recent development of machine learning-based methods for the prediction of defensin family and subfamily https://www.excli.de/index.php/excli/article/view/4913 <p>Nearly all living species comprise of host defense peptides called defensins, that are crucial for innate immunity. These peptides work by activating the immune system which kills the microbes directly or indirectly, thus providing protection to the host. Thus far, numerous preclinical and clinical trials for peptide-based drugs are currently being evaluated. Although, experimental methods can help to precisely identify the defensin peptide family and subfamily, these approaches are often time-consuming and cost-ineffective. On the other hand, machine learning (ML) methods are able to effectively employ protein sequence information without the knowledge of a protein’s three-dimensional structure, thus highlighting their predictive ability for the large-scale identification. To date, several ML methods have been developed for the <em>in silico</em> identification of the defensin peptide family and subfamily. Therefore, summarizing the advantages and disadvantages of the existing methods is urgently needed in order to provide useful suggestions for the development and improvement of new computational models for the identification of the defensin peptide family and subfamily. With this goal in mind, we first provide a comprehensive survey on a collection of six state-of-the-art computational approaches for predicting the defensin peptide family and subfamily. Herein, we cover different important aspects, including the dataset quality, feature encoding methods, feature selection schemes, ML algorithms, cross-validation methods and web server availability/usability. Moreover, we provide our thoughts on the limitations of existing methods and future perspectives for improving the prediction performance and model interpretability. The insights and suggestions gained from this review are anticipated to serve as a valuable guidance for researchers for the development of more robust and useful predictors.</p> Phasit Charoenkwan Nalini Schaduangrat S. M. Hasan Mahmud Orawit Thinnukool Watshara Shoombuatong Copyright (c) 2022 Phasit Charoenkwan, Nalini Schaduangrat, S. M. Hasan Mahmud, Orawit Thinnukool, Watshara Shoombuatong http://creativecommons.org/licenses/by/4.0 2022-05-05 2022-05-05 21 757 771 10.17179/excli2022-4913 Systems redox biology in health and disease https://www.excli.de/index.php/excli/article/view/4793 <p class="Abstract"><span lang="EN-US">Living organisms need to be able to cope with environmental challenges and other stressors and mount adequate responses that are as varied as the spectrum of those challenges. Understanding how the multi-layered biological stress responses become integrated across and between different levels of organization within an organism can provide a different perspective on the nature and inter-relationship of complex systems in health and disease. We here compare two concepts which have been very influential in stress research: Selye’s ‘<em>General Adaptation Syndrome</em>’ and Sies’s ‘<em>Oxidative Stress</em>’ paradigm. We show that both can be embraced within a more general framework of ‘change and response’. The ‘<em>Reactive Species Interactome</em>’ allows each of these to be considered as distinct but complementary aspects of the same system, representative of roles at different levels of organization within a functional hierarchy. The versatile chemistry of sulfur - exemplified by hydrogen sulfide, glutathione and proteinous cysteine thiols - enriched by its interactions with reactive oxygen, nitrogen and sulfur species, would seem to sit at the heart of the ‘Redox Code’ and underpin the ability of complex organisms to cope with stress.</span></p> Martin Feelisch Miriam M. Cortese-Krott Jérôme Santolini Stephen A. Wootton Alan A. Jackson Copyright (c) 2022 Martin Feelisch, Miriam M. Cortese-Krott, Jerome Santolini, Stephen A. Wootton, Alan A. Jackson http://creativecommons.org/licenses/by/4.0 2022-03-21 2022-03-21 21 623 646 10.17179/excli2022-4793 Empirical comparison and analysis of machine learning-based predictors for predicting and analyzing of thermophilic proteins https://www.excli.de/index.php/excli/article/view/4723 <p>Thermophilic proteins (TPPs) are critical for basic research and in the food industry due to their ability to maintain a thermodynamically stable fold at extremely high temperatures. Thus, the expeditious identification of novel TPPs through computational models from protein sequences is very desirable. Over the last few decades, a number of computational methods, especially machine learning (ML)-based methods, for <em>in silico</em> prediction of TPPs have been developed. Therefore, it is desirable to revisit these methods and summarize their advantages and disadvantages in order to further develop new computational approaches to achieve more accurate and improved prediction of TPPs. With this goal in mind, we comprehensively investigate a large collection of fourteen state-of-the-art TPP predictors in terms of their dataset size, feature encoding schemes, feature selection strategies, ML algorithms, evaluation strategies and web server/software usability. To the best of our knowledge, this article represents the first comprehensive review on the development of ML-based methods for in <em>silico</em> prediction of TPPs. Among these TPP predictors, they can be classified into two groups according to the interpretability of ML algorithms employed (i.e., computational black-box methods and computational white-box methods). In order to perform the comparative analysis, we conducted a comparative study on several currently available TPP predictors based on two benchmark datasets. Finally, we provide future perspectives for the design and development of new computational models for TPP prediction. We hope that this comprehensive review will facilitate researchers in selecting an appropriate TPP predictor that is the most suitable one to deal with their purposes and provide useful perspectives for the development of more effective and accurate TPP predictors.</p> Phasit Charoenkwan Nalini Schaduangrat Md Mehedi Hasan Mohammad Ali Moni Pietro Lió Watshara Shoombuatong Copyright (c) 2022 Phasit Charoenkwan, Nalini Schaduangrat, Md Mehedi Hasan, Mohammad Ali Moni, Pietro Lió, Watshara Shoombuatong http://creativecommons.org/licenses/by/4.0 2022-03-02 2022-03-02 21 554 570 10.17179/excli2022-4723 The effect of exercise interventions on Irisin level https://www.excli.de/index.php/excli/article/view/4703 <p>Irisin is a hormone that is offered to be a hopeful remedial target in obesity and type 2 diabetes. It has received striking attention recently, whereas, the interactions between exercise training and irisin are still unclear. Therefore, this systematic review and meta-analysis investigated the impacts of exercise interventions on circulating irisin in adults. A systematic search was conducted in PubMed, CINAHL, MEDLINE, Cochrane, Google Scholar, and Scopus up to July 15, 2021. Twenty-four studies, which assessed a total of 921 participants were included and analyzed using a random-effects model to estimate weighted mean differences (MD) with 95 % confidence intervals (CI). Overall, data revealed that exercise training significantly increased circulating irisin (MD: 0.01, 95 % CI: 0.00, 0.01, p = 0.005), and declined insulin (MD: -2.09, 95 % CI: -2.81, -1.37, p &lt; 0.00001), glucose (MD: -12.89, 95 % CI: -16.52, -9.26, p &lt; 0.00001), and insulin resistance (MD: -0.89, 95 % CI: -1.15, -0.62, p &lt; 0.00001). Subgroup analysis revealed that irisin raised significantly when resistance training (p = 0.04) and combined training (p = 0.002) were applied, and for the type 2 diabetes and prediabetes (p = 0.002 for both) groups. Moreover, subgroup analysis by the type of intervention demonstrated that insulin reduced when aerobic training (p &lt; 0.00001) and combined training (p = 0.0003) were employed, but glucose and HOMA-IR reduced after all three types of exercise training. These findings demonstrate that exercise interventions may produce ameliorations in circulating irisin. Further long-term studies are required to confirm these findings.</p> Gholam Rasul Mohammad Rahimi Keyvan Hejazi Martin Hofmeister Copyright (c) 2022 Gholam Rasul Mohammad Rahimi, Keyvan Hejazi, Martin Hofmeister http://creativecommons.org/licenses/by/4.0 2022-02-25 2022-02-25 21 524 539 10.17179/excli2022-4703 Oxidative stress and visual system https://www.excli.de/index.php/excli/article/view/4663 <p class="Abstract"><span lang="EN-US">Different types of tissues respond differently to the action of oxidative stress. The visual system is very sensitive to oxidative action due to continuous exposure to light. In consideration of the growing interest of scientific studies towards various compounds endowed with antioxidant and anti-inflammatory properties, we performed a review of the literature focusing on the use of some antioxidant molecules for the treatment of conditions affecting the visual system. In this study, we focused on the ability of two antioxidant agents, the small molecule α-lipoic acid (ALA) and the enzyme superoxide dismutase (SOD), to influence the neurodegenerative physiological processes related to aging and oxidative stress affecting the ocular segment. The literature data report that ALA and SOD can protect against neurodegenerative effects both the optic nerve and retina and, if administered together, they are able to lower the levels of oxidative stress, thus preventing neurodegeneration and reducing the apoptotic process.</span></p> Samanta Taurone Massimo Ralli Marco Artico Valentina Noemi Madia Susanna Scarpa Stefania Annarita Nottola Antonio Maconi Marta Betti Pietro Familiari Marcella Nebbioso Roberta Costi Alessandra Micera Copyright (c) 2022 Samanta Taurone, Massimo Ralli, Marco Artico, Valentina Noemi Madia, Susanna Scarpa, Stefania Annarita Nottola, Antonio Maconi, Marta Betti, Pietro Familiari, Marcella Nebbioso, Roberta Costi, Alessandra Micera http://creativecommons.org/licenses/by/4.0 2022-03-01 2022-03-01 21 544 553 10.17179/excli2022-4663 Quantitative aspects of nitric oxide production from nitrate and nitrite https://www.excli.de/index.php/excli/article/view/4727 <p>Nitric oxide (NO) is involved in many physiological and pathological processes in the human body. At least two major pathways produce NO: (1) the <em>L</em>-arginine-NO-oxidative pathway in which NO synthase (NOS) enzymes convert <em>L</em>-arginine to NO; (2) the nitrate-nitrite-NO reductive pathway in which NO is produced from the serial reduction of nitrate and nitrite. The deficiency of NO is involved in the pathophysiology of cardiometabolic disorders. Intervention with foods containing nitrate and nitrite can potentially prevent or treat some chronic diseases, including cardiovascular diseases and diabetes. A better understanding of the NO cycle would help develop effective strategies for preventing or treating the disorders in which NO homeostasis is disturbed. This review summarizes quantitative aspects of NO production, emphasizing the nitrate-nitrite-NO pathway. Available data indicates that total NO production by NOS-dependent <em>L</em>-arginine-NO pathway is about 1000 μmol.day<sup>-1</sup>. Of about 1700 μmol.day<sup>-1 </sup>ingested nitrate, ~25 % is extracted by the salivary glands and of which ~20 % is converted nitrite. It means that about 5 % of ingested nitrate is converted to nitrite in the oral cavity; assuming that all produced nitrite is reduced to NO in the stomach, it can be calculated that contribution of the nitrate-nitrite-NO pathway to the whole-body NO production is about 85 μmol.day<sup>-1 </sup>(1700 ×0.05=85) or approximately 100 μmol.day<sup>-1</sup>. The lower contribution of the nitrate-nitrite-NO pathway does not mean that this pathway has lower importance in the whole-body NO homeostasis. Even in the adequate <em>L</em>-arginine supply, NOS-dependent NO production is insufficient to meet all NO functions, and the nitrate-nitrite-NO pathway must provide the rest. In conclusion, the contribution of the nitrate-nitrite-NO pathway in the whole human body NO production is &lt;10 %, and the nitrate-nitrite-NO pathway is complementary to the NOS-dependent NO production.</p> Asghar Ghasemi Copyright (c) 2022 Asghar Ghasemi http://creativecommons.org/licenses/by/4.0 2022-02-21 2022-02-21 21 470 486 10.17179/excli2022-4727 Evaluating the adverse outcome of subtypes of heart failure with preserved ejection fraction defined by machine learning https://www.excli.de/index.php/excli/article/view/4572 <p>The ability to distinguish clinically meaningful subtypes of heart failure with preserved ejection fraction (HFpEF) has recently been examined by machine learning techniques but studies appear to have produced discordant results. The objective of this study is to synthesize the types of HFpEF by examining their features and relating them to phenotypes with adverse prognosis. A systematic search was conducted using the search terms “Diastolic Heart Failure” OR “heart failure with preserved ejection fraction” OR “heart failure with normal ejection fraction” OR “HFpEF” AND “machine learning” OR “artificial intelligence” OR ‘computational biology’. Ten studies were identified and they varied in their prevalence of ten clinical variables: age, sex, body mass index (BMI) or obesity, hypertension, diabetes mellitus, coronary artery disease, atrial fibrillation, chronic kidney disease, chronic obstructive pulmonary disease or symptom severity (NYHA class or BNP). The clinical findings associated with the different phenotypes in &gt; 85 % of studies were age, hypertension, atrial fibrillation, chronic kidney disease and worse symptoms severity; an adverse outcome was in 65 % to 85 % of studies identified diabetes mellitus and female sex and in less than 65 % of studies was body mass index or obesity, and coronary artery disease. COPD was a relevant factor in only 33 % of studies. Adverse clinical outcome – death or admission to hospital (for heart failure) defined phenogroups with the worst outcome. Combining the 4 studies that calculated the MAGGIC score showed a significant (p&lt;0.05) linear relationship between MAGGIC score and outcome, using the one-year event rate. A new score based on strength of the evidence of the HFpEF studies analyzed here, using 9 variables (eliminating COPD), showed a significant (p&lt;0.009) linear relationship with one-year event rate. Three studies examined biomarkers in detail and the ones most prominently related to outcome or consistently found in the studies were GDF15, FABP4, FGF23, sST2, renin and TNF. The dominant factors that identified phenotypes of HFpEF with adverse outcome were hypertension, atrial fibrillation, chronic kidney disease and worse symptoms severity. A new simplified score, based on clinical factors, was proposed to assess prognosis in HFpEF. Several biomarkers were consistently elevated in phenogroups with adverse outcomes and may indicate the underlying mechanism or pathophysiology specific for phenotypes with an adverse prognosis.</p> Simon W. Rabkin Copyright (c) 2022 Simon W. Rabkin http://creativecommons.org/licenses/by/4.0 2022-02-22 2022-02-22 21 487 518 10.17179/excli2021-4572 Molecular mechanisms linking stress and insulin resistance https://www.excli.de/index.php/excli/article/view/4382 <p class="Abstract"><span lang="EN-US">To date, there is ample evidence to support the strong relationship between stress and insulin resistance. While diabetes mellitus acts as a potent stress inducer, stress may be an upstream event for insulin resistance as well. It is widely recognized that diabetes mellitus is more prevalent among people who have a stressful lifestyle; however, the underlying mechanisms are not well understood. In the current study, we surveyed the scientific literature for possible interactions between stress and insulin resistance and found that stress can impair glucose homeostasis, working through at least six molecular pathways. </span></p> Habib Yaribeygi Mina Maleki Alexandra E. Butler Tannaz Jamialahmadi Amirhossein Sahebkar Copyright (c) 2022 Habib Yaribeygi, Mina Maleki, Alexandra E. Butler, Tannaz Jamialahmadi, Amirhossein Sahebkar http://creativecommons.org/licenses/by/4.0 2022-01-24 2022-01-24 21 317 334 10.17179/excli2021-4382 “Nano-ghosts”: Risk assessment of submicron-sized particles in food biased towards fictional “nano” https://www.excli.de/index.php/excli/article/view/4630 <p>Much confusion has been generated in the safety assessment of food-grade TiO<sub>2</sub> (E171) by the comingling of studies conducted on submicron-sized particles with those examining the toxicity of more minuscule counterparts. As E171 displays a nano-sized tail in its particle distribution (up to 36 % of particles with a diameter &lt; 100 nm), it was thought that potential hazards of this food additive can be extrapolated from studies on thoroughly nanoscale formulations. This simplistic procedure may, however, overestimate the effects of the nano-sized tail of E171 because TiO<sub>2</sub> particles readily aggregate or agglomerate in aqueous suspensions and biological matrices. The resulting larger clusters display a reduced oral bioavailability in comparison to the same material in nano-sized dimensions. Also, even if taken up in trace amounts, the smaller particles likely remain appended to larger particles or clusters and these aggregates or conglomerates may nullify to a great extent their “nano” characteristics. The purpose of this review is, therefore, to reevaluate the literature on the toxicity of TiO<sub>2</sub> particles focusing on studies that are directly relevant for the assessment of E171. The purpose is not to avert a ban on the use of E171 in food, which might well be justified in light of the uncertainties associated with this additive employed solely for its colorant properties. Instead, it will be important to avoid in the future this same bias towards a fictional “nano” hazard, especially when evaluating more innovative engineered particles that confer true benefits for example by enhancing nutritional properties, quality, freshness, traceability or sustainability of food.</p> Hanspeter Naegeli Corina Gsell Copyright (c) 2022 Hanspeter Naegeli, Corina Gsell http://creativecommons.org/licenses/by/4.0 2022-01-13 2022-01-13 21 279 299 10.17179/excli2022-4630 LOX-1: Implications in atherosclerosis and myocardial ischemia https://www.excli.de/index.php/excli/article/view/4532 <p class="Abstract"><span lang="EN-US">Understanding the pathophysiology of atherosclerosis is fundamental to the practice of cardiovascular medicine. Atherosclerosis is a multi-step cascade of accumulation of lipids and downstream changes that lead to a fibro-fatty plaque formation in the arterial intima. Multiple biochemical stimuli, cellular receptors and intra-cellular signals are implicated in this complex mechanism. Lectin-type oxidized LDL receptor-1 or LOX-1 is a type II membrane glycoprotein receptor which has emerged as an important effector of atherosclerosis. Hence, LOX-1 modification and its clinical consequences are of much interest in recent times.</span></p> Tanya Sharma Francesco Romeo Jawahar L. Mehta Copyright (c) 2022 Tanya Sharma, Francesco Romeo, Jawahar L. Mehta http://creativecommons.org/licenses/by/4.0 2022-01-13 2022-01-13 21 273 278 10.17179/excli2021-4532 Prognostic significance and therapeutic potentials of immune checkpoints in osteosarcoma https://www.excli.de/index.php/excli/article/view/4094 <p class="Abstract"><span lang="EN-US">Although there exist manifold strategies for cancer treatment, researchers are obliged to develop novel treatments based on the challenges that arise. One of these recent treatment approaches is cancer immunotherapy, which enjoys various types of strategies itself. However, one of the most significant methods, in this regard, is employing immune checkpoint proteins (ICPs). Bone sarcomas have several subtypes, with the most common ones being chordoma, chondrosarcoma, Ewing sarcoma, and osteosarcoma. Although many aggressive treatment approaches, including radiotherapy, chemotherapy, and surgical resection, have been employed over the last decades, significantly improved outcomes have not been observed for Ewing sarcoma or osteosarcoma patients. Additionally, chordoma and chdrosarcoma resist against both radiation and chemotherapy. Accordingly, elucidating how recent therapies could affect bone sarcomas is necessary. Checkpoint inhibitors have attracted great attention for the treatment of several cancer types, including bone sarcoma. Herein, the recent advances of current immune checkpoint targets, such as anti-PD-1/PD-L1 and anti-CTLA-4 blockade, for the treatment of bone sarcoma have been reviewed. </span></p> Vafa Meftahpour Ali Aghebati-Maleki Ali Fotouhi Elham Safarzadeh Leili Aghebati-Maleki Copyright (c) 2022 Vafa Meftahpour, Ali Aghebati-Maleki, Ali Fotouhi, Elham Safarzadeh, Leili Aghebati-Maleki http://creativecommons.org/licenses/by/4.0 2022-01-12 2022-01-12 21 250 268 10.17179/excli2021-4094 The effects of low-toxic herbicide Roundup and glyphosate on mitochondria https://www.excli.de/index.php/excli/article/view/4478 <p>The effects of pesticides on the health of non-target living organisms in agricultural areas are critically important aspects for their safe use. Their release into the environment is an inevitable aspect for predicting and evaluation of the risk of their application. Roundup, a glyphosate-based herbicide, has been designed as an effective pesticide against weeds and now is the most widely used agrochemicals around the world due to its highly specific action of the biosynthesis of certain amino acids in plants. Despite it is claimed to be low toxic for not-target organisms, due to its broad application Roundup and products of its degradation were detected in organisms of diverse animals and humans. In this review, we describe animal and human studies of general adverse effects of Roundup and its principal substance glyphosate with focus on endocrine disruption, oxidative stress and behavioral disorders. At mechanistic level, we focus on the potential toxicity of the herbicide Roundup and glyphosate as effectors of bioenergetic functions of mitochondria. Their effects on mitochondrial membrane potential and oxidative phosphorylation are among described to date critical components responsible for its toxicity. Finally, we discuss general molecular mechanisms potentially involved in the interaction between glyphosate and mitochondria which to some extent are associated with generation of reactive oxygen species.</p> Olha M. Strilbytska Sviatoslav A. Tsiumpala Ivanna I. Kozachyshyn Tetiana Strutynska Nadia Burdyliuk Volodymyr Lushchak Oleh Lushchak Copyright (c) 2022 Olha M. Strilbytska, Sviatoslav A. Tsiumpala, Ivanna I. Kozachyshyn, Tetiana Strutynska, Nadia Burdyliuk, Volodymyr Lushchak, Oleh Lushchak http://creativecommons.org/licenses/by/4.0 2022-01-10 2022-01-10 21 183 196 10.17179/excli2021-4478 Epidemiology and biology of early onset colorectal cancer https://www.excli.de/index.php/excli/article/view/4456 <p>Colorectal cancer (CRC) is the third leading cause of cancer-related mortality in men or women in the United States. Average-risk screening that begins at age 50 years has reduced incidence and mortality of CRC in those over 50 years of age, whereas CRC incidence in those under age 50 years (early onset colorectal cancer (eoCRC)) has recently and dramatically increased. In this review, we summarize the recent literature including risk factors for eoCRC, differences in clinicopathologic presentation and outcomes in eoCRC, and emerging evidence regarding the molecular pathways that are altered in eoCRC compared to later onset CRC (loCRC). Epidemiologic studies of eoCRC show predominance in distal colon and rectum, and association with several modifiable risk factors, including diabetes, obesity, diet, sedentary time, alcohol consumption and smoking. Data regarding potential risk factors of prior antibiotic exposure and microbiome alterations or direct carcinogen exposure are still emerging. Aggressive clinicopathologic features of eoCRC at presentation may be due to delay in diagnosis or more aggressive tumor biology. EoCRC outcomes are similar to loCRC when matched for stage, but overall mortality is greater due to higher frequency of advanced disease at a younger presentation, with more life-years lost. There are only few molecular evaluations of eoCRC to date, with findings of potential increase in <em>TP53</em> and <em>CTNNB1</em> somatic mutation and decrease in <em>APC</em>, <em>KRAS</em> and <em>BRAF</em> somatic mutation, compared to loCRC. Other findings include LINE-1 hypomethylation, absence of microsatellite instability (MSI-H), presence of chromosomal instability (CIN) or microsatellite and chromosomal stability (MACS). These studies are only now emerging and have not yet identified a specific molecular signature defining eoCRC. Further research evaluating genetic and molecular differences as well as environmental triggers for eoCRCs should provide a clearer understanding to inform targeted screening for pre-symptomatic at-risk younger individuals.</p> Anand Venugopal John M. Carethers Copyright (c) 2022 Anand Venugopal, John M. Carethers http://creativecommons.org/licenses/by/4.0 2022-01-07 2022-01-07 21 162 182 10.17179/excli2021-4456 Potential dual inhibitors of PCSK-9 and HMG-R from natural sources in cardiovascular risk management https://www.excli.de/index.php/excli/article/view/4453 <p class="Abstract"><span lang="EN-US">Atherosclerotic cardiovascular disease (ASCVD) stands amongst the leading causes of mortality worldwide and has attracted the attention of world’s leading pharmaceutical companies in order to tackle such mortalities. The low-density lipoprotein-cholesterol (LDL-C) is considered the most prominent biomarker for the assessment of ASCVD risk. Distinct inhibitors of 3-hydroxy-3-methyl-glutaryl-CoA reductase (HMG-R), the chief hepatic cholesterogenic enzyme, are being used since last seven decades to manage hypercholesterolemia. On the other hand, discovery and the association of proprotein convertase subtilisin/kexin type-9 (PCSK-9) with increased ASCVD risk have established PCSK-9 as a novel therapeutic target in cardiovascular medicine. PCSK-9 is well reckoned to facilitate the LDL-receptor (LDL-R) degradation and compromised LDL-C clearance leading to the arterial atherosclerotic plaque formation. The currently available HMG-R inhibitors (statins) and PCSK-9 inhibitors (siRNA, anti-sense oligonucleotides, and monoclonal antibodies) have shown great promises in achieving LDL-C lowering goals, however, their life long prescriptions have raised significant concerns. These deficits associated with the synthetic HMG-R and PCSK-9 inhibitors called for the discovery of alternative therapeutic candidates with potential dual HMG-R and PCSK-9 inhibitory activities from natural origins. Therefore, this report firstly describes the mechanistic insights into the cholesterol homeostasis through HMG-R, PCSK-9, and LDL-R functionality and then compiles the pharmacological effects of natural secondary metabolites with special emphasis on their dual HMG-R and PCSK-9 inhibitory action. In conclusion, various natural products exhibit atheroprotective effects via targeting HMG-R and PCSK-9 activities and lipoprotein metabolism, however, further clinical assessments are still warranted prior their approval for ASCVD risk management in hypercholesterolemic patients.</span></p> Mohd Waiz Sahir Sultan Alvi M. Salman Khan Copyright (c) 2022 Mohd Waiz, Sahir Sultan Alvi, M. Salman Khan http://creativecommons.org/licenses/by/4.0 2022-01-05 2022-01-05 21 47 76 10.17179/excli2021-4453 Large-scale comparative review and assessment of computational methods for phage virion proteins identification https://www.excli.de/index.php/excli/article/view/4411 <p>Phage virion proteins (PVPs) are effective at recognizing and binding to host cell receptors while having no deleterious effects on human or animal cells. Understanding their functional mechanisms is regarded as a critical goal that will aid in rational antibacterial drug discovery and development. Although high-throughput experimental methods for identifying PVPs are considered the gold standard for exploring crucial PVP features, these procedures are frequently time-consuming and labor-intensive. Thusfar, more than ten sequence-based predictors have been established for the <em>in silico</em> identification of PVPs in conjunction with traditional experimental approaches. As a result, a revised and more thorough assessment is extremely desirable. With this purpose in mind, we first conduct a thorough survey and evaluation of a vast array of 13 state-of-the-art PVP predictors. Among these PVP predictors, they can be classified into three groups according to the types of machine learning (ML) algorithms employed (i.e. traditional ML-based methods, ensemble-based methods and deep learning-based methods). Subsequently, we explored which factors are important for building more accurate and stable predictors and this included training/independent datasets, feature encoding algorithms, feature selection methods, core algorithms, performance evaluation metrics/strategies and web servers. Finally, we provide insights and future perspectives for the design and development of new and more effective computational approaches for the detection and characterization of PVPs.</p> Muhammad Kabir Chanin Nantasenamat Sakawrat Kanthawong Phasit Charoenkwan Watshara Shoombuatong Copyright (c) 2022 Muhammad Kabir , Chanin Nantasenamat, Sakawrat Kanthawong, Phasit Charoenkwan, Watshara Shoombuatong http://creativecommons.org/licenses/by/4.0 2022-01-03 2022-01-03 21 11 29 10.17179/excli2021-4411 Unilateral hand force control impairments in older women https://www.excli.de/index.php/excli/article/view/5362 <p>Older women may experience deficits in sensorimotor control at their upper limb because of aging progress compromising the motor system. This study aimed to investigate whether younger and older women differ in sensorimotor capabilities assessed by unilateral force control performances at a lower targeted force level. Twenty-one older and 21 younger women performed isometric unilateral force control tasks at 10 % of maximum voluntary contraction for each hand, respectively. Purdue Pegboard Test (PPT) was used to measure unilateral hand dexterity. Five force control variables (i.e., maximal and submaximal force, force error, variability, and regularity) and PPT scores were analyzed in two-way mixed ANOVAs (Group × Hand Condition), respectively. The absolute force power was analyzed in three-way mixed ANOVA (Group × Hand Condition × Frequency Band). The findings revealed that older women produced less maximal and submaximal unilateral forces than in younger women. Greater variability, regularity, and force frequency oscillations below 4 Hz were observed in older women as compared with those in younger women. Force error in the dominant hand was greater in older women than those in younger women. Finally, older women showed lower PPT scores than younger women. These findings suggested that older women may have deficits in unilateral force control capabilities as well as motor dexterity.</p> Hanall Lee Young-Min Park Nyeonju Kang Copyright (c) 2022 Hanall Lee, Young-Min Park, Nyeonju Kang http://creativecommons.org/licenses/by/4.0 2022-09-22 2022-09-22 21 1231 1244 10.17179/excli2022-5362 Intraspecific differences in Androctunus crassicauda venom and envenomation symptoms https://www.excli.de/index.php/excli/article/view/5078 <p>Envenomation by <em>Androctunus crassicauda</em> is very frequent in Iran, especially in the south-west<em>. </em>This scorpion is one of the six scorpions whose venom is used to prepare anti-venom. Using HPLC, we discovered venom components of <em>A. crassicauda</em> varies from one specimen to another depending on geographical location, and this result is confirmed by those first found in various symptoms of <em>A. crassicauda</em> sting in envenomed persons from two separate geographical places (north and south of Khuzestan province). There was a significant relationship between symptoms and location of envenomation by <em>A. crassicauda</em>. Muscle spasm was more dominant in envenomed people from Northern cities, and venom chromatogram analysis showed the presence of at least six main sharp peaks in Northern <em>A. crassicauda</em> rather than Southern<em> A. crassicauda</em>. It shows intraspecific differences in venom of <em>A. crassicauda</em> that must be considered in treatment of stung people from different geographical locations as well as in the preparation of anti-venom.</p> Mehdi Safdarian Babak Vazirianzadeh Ahmad Ghorbani Narges Pashmforoosh Masoumeh Baradaran Copyright (c) 2022 Mehdi Safdarian, Babak Vazirianzadeh, Ahmad Ghorbani, Narges Pashmforoosh, Masoumeh Baradaran http://creativecommons.org/licenses/by/4.0 2022-09-21 2022-09-21 21 1222 1230 10.17179/excli2022-5078 Correlation of miR-24-3p and miR-595 expression with CCL3, CCL4, IL1-β, TNFαIP3, and NF-κBIα genes in PBMCs of patients with coronary artery disease https://www.excli.de/index.php/excli/article/view/5266 <p>Inflammation has been well recognized to play an important role in developing coronary artery disease (CAD). By regulating essential genes in this pathway post-transcriptionally, MicroRNAs (miRNAs) may help or hinder the development of atherosclerotic lesions. The aim of this study was to investigate the expression of miR-24-3p, miR-595, <em>CCL3</em>, <em>CCL4</em>, <em>IL-1β</em>, <em>TNFαIP3</em>, and <em>NF-κBIα</em> in the peripheral blood mononuclear cells (PBMCs) of CAD and control groups and to examine whether any correlation exists between the expression of miRs and genes in CAD group. A total of 168 subjects (84 CAD subjects and 84 control subjects) were examined in this research. Expression levels of miR-24-3p, miR-595, CCL3, CCL4, IL-1β<em>, TNFαIP3</em>, and <em>NF-κBIα</em> in PBMCs were measured using the real-time PCR technique. A comparison of the CAD group with the control group indicated significantly increased expression levels of <em>CCL3</em>, <em>CCL4,</em> and <em>IL-1β </em>(Fold Change (FC)=4, <em>P</em>=0.009; FC=2.9, <em>P</em>=0.01; FC=1.8, <em>P</em>=0.019, respectively) and remarkably reduced expression levels of <em>TNFαIP3</em> and <em>NF-κBIα </em>(FC=-1.4, <em>P</em>=0.03 and FC=-5.9, <em>P</em>=0.001, respectively). Moreover, the expression levels of miR-24-3p downregulated (FC=-2.5, <em>P</em>=0.005) and miR-595 upregulated (FC=1.9, <em>P</em>=0.009) in the CAD group. There was a statistical correlation between the number of clogged arteries with expression levels of miR-24-3p, miR-595, <em>CCL3</em>, <em>CCL4</em>, <em>IL-1β</em>, <em>TNFαIP3</em>, and <em>NF-κBIα</em> in the CAD group. Also, there was a statistical correlation between expression levels of miR-24-3p and miR-595 with <em>CCL3</em>, <em>CCL4</em>, <em>IL-1β</em>, <em>TNFαIP3</em>, and <em>NF-κBIα</em> gene expression in the CAD group. In CAD patients, decreased expression of miR-24-3p and increased expression of miR-595 may aid the progression of atherosclerotic plaques by regulating <em>CCL3</em>, <em>CCL4</em>, <em>IL-1β</em>, <em>TNFαIP3</em>, and <em>NF-κBIα</em> gene expression.</p> Shakiba Kazemian Reza Ahmadi Gordon A. Ferns Ali Rafiei Fatemeh Azadegan-Dehkordi Arsalan Khaledifar Mina Mohammad-Rezaei Nader Bagheri Copyright (c) 2022 Shakiba Kazemian, Reza Ahmadi, Gordon A. Ferns, Ali Rafiei, Fatemeh Azadegan-Dehkordi, Arsalan Khaledifar, Mina Mohammad-Rezaei, Nader Bagheri http://creativecommons.org/licenses/by/4.0 2022-09-08 2022-09-08 21 1184 1195 10.17179/excli2022-5266 Association between expression of ZBP1, AIM2, and MDA5 genes and severity of COVID-19 https://www.excli.de/index.php/excli/article/view/5141 <p class="Abstract"><span lang="EN-US">Antiviral and inflammatory responses following the detection of the virus genome by nucleic acid sensors play a vital role in the pathogenesis and outcome of diseases. In this study, we investigated the ZBP<span style="color: black;">1, </span>AIM2,<span style="color: black;"> and MDA5 </span>expression levels in COVID-19 patients with different intensities of the disease. 75 quantitative Real-Time PCR (qRT-PCR)-confirmed COVID-19 patients were included consecutively and divided into 3 groups of mild, severe, and critical based on the severity of the disease. Also, 25 healthy volunteer subjects were included. PBMCs were collected from the whole blood, and RNA was extracted using commercial kit. The expression of ZBP1, AIM2, and MDA5 genes was investigated using qRT-PCR technique. The mean age of the patients and healthy volunteers was 52.73±13.78 and 49.120±12.490, respectively. In each group, 13 out of 25 participants were male. The expression levels of ZBP1 (P=0.001), AIM2 (P=0.001), and MDA5 (P= 0.003) transcript were significantly higher in COVID-19 patients than the control group. <a name="_Hlk98580051"></a>The results also revealed that the expression levels of ZBP1, AIM2, and MDA5 were significantly higher in the critical and severe COVID-19 patients compared to those with mild disease (P&lt;0.05). Moreover, regarding the gender, the expression levels of AIM2 and MDA5 were significantly elevated in male severe (P=0.04 and P=0.003, respectively) and critical (P=0.005 and P=0.0004, respectively) patients than the female ones. The results indicated that ZBP1, AIM2, and MDA5 genes might have an important role in the severity of COVID-19 disease. Moreover, the severity of COVID-19 disease in male and female patients might be related to AIM2, and MDA5 expression levels. More studies are recommended to be conducted to clarify this issue. </span></p> Nasir Arefinia Amin Ramezani Mehrdad Farokhnia Ali Mohammad Arab Zadeh Ramin Yaghobi Jamal Sarvari Copyright (c) 2022 Nasir Arefinia, Amin Ramezani, Mehrdad Farokhnia, Ali Mohammad Arab Zadeh, Ramin Yaghobi, Jamal Sarvari http://creativecommons.org/licenses/by/4.0 2022-09-01 2022-09-01 21 1171 1183 10.17179/excli2022-5141 Long-term inorganic nitrate administration protects against ovariectomy-induced osteoporosis in rats https://www.excli.de/index.php/excli/article/view/5082 <p>The risk of osteoporotic fractures increases in women after menopause. This study aims at determining the effects of long-term inorganic nitrate administration against ovariectomy-induced osteoporosis in rats. Rats were divided into 4 groups (n=6/group): Control, control+nitrate, ovariectomized (OVX), and OVX+nitrate. Sodium nitrate (100 mg/L in drinking water) was administered for 9 months. Trabecular bone quality in the proximal tibia was measured using a Micro-Computed Tomography (micro-CT) scanner at months 0, 1, 3, and 9. Levels of nitric oxide (NO) metabolites (NOx) and oxidative stress indices, and mRNA expression of endothelial NO synthase (eNOS) were measured at month 9 in the proximal tibia. Compared to controls, OVX rats had lower NOx levels by 47 %, eNOS mRNA expression by 55 %, catalase activity (CAT) by 45 %, total antioxidant capacity (TAC) by 70 %, and higher malondialdehyde (MDA) levels by 327 % in the bone tissue at month 9. OVX rats, compared to controls, had lower bone volume/tissue volume (BV/TV), trabecular number (Tb.N.), and trabecular thickness (Tb.Th.) by 32 %, 58 %, and 17 %, respectively, and higher trabecular separation (Tb.Sp.) by 123 %, at month 9. Nitrate administration to control rats increased TAC by 46 % in the bone tissue at month 9 but did not significantly affect other parameters in serum and bone tissue. Nitrate in OVX rats significantly increased NOx levels by 86 %, eNOS expression by 2.14-fold, CAT activity by 75 %, TAC by 170 %, and decreased MDA levels by 36 % at month 9 in the bone tissue. Nitrate-treated OVX rats at month 9 had higher BV/TV (42 %) and Tb.N. (61 %) and lower Tb.Sp. (15 %). Long-term inorganic nitrate administration at a low dose has protective effects against OVX-induced osteoporosis in rats; this effect is associated with increasing eNOS-derived NO and decreasing oxidative stress in the bone tissue.</p> Nasibeh Yousefzadeh Sajad Jeddi Khosrow Kashfi Asghar Ghasemi Copyright (c) 2022 Nasibeh Yousefzadeh, Sajad Jeddi, Khosrow Kashfi, Asghar Ghasemi http://creativecommons.org/licenses/by/4.0 2022-08-24 2022-08-24 21 1151 1166 10.17179/excli2022-5082 Expression of connexins and pannexins in diseased human liver https://www.excli.de/index.php/excli/article/view/5163 <p class="Abstract"><span lang="EN-US">Connexin proteins can form hexameric hemichannels and gap junctions that mediate paracrine and direct intercellular communication, respectively. Gap junction activity is crucial for the maintenance of hepatic homeostasis, while connexin hemichannels become particularly active in liver disease, such as hepatitis, fibrosis, cholestasis or even hepatocellular carcinoma. Channels consisting of connexin-like proteins named pannexins have been directly linked to liver inflammation and cell death. The goal of the present study was to characterize the expression and subcellular localization of connexins and pannexins in liver of patients suffering from various chronic and neoplastic liver diseases. Specifically, real-time quantitative reverse transcription polymerase chain reaction, immunoblotting and immunohistochemistry analyses were performed on human liver biopsies. It was found that pannexin1 and pannexin2 gene expression are correlated to a certain degree, as is pannexin1 protein expression with connexin32 and connexin43 protein expression. Furthermore, this study is the first to detect pannexin3 in human patient liver biopsies <em>via</em> both immunoblot and immunohistochemistry.</span></p> Kaat Leroy Vânia Vilas-Boas Eva Gijbels Bart Vanderborght Lindsey Devisscher Bruno Cogliati Bert Van Den Bossche Isabelle Colle Mathieu Vinken Copyright (c) 2022 Kaat Leroy, Vânia Vilas-Boas, Eva Gijbels, Bart Vanderborght, Lindsey Devisscher, Bruno Cogliati, Bert Van Den Bossche, Isabelle Colle, Mathieu Vinken http://creativecommons.org/licenses/by/4.0 2022-08-22 2022-08-22 21 1111 1129 10.17179/excli2022-5163 COVID-19 lockdown https://www.excli.de/index.php/excli/article/view/4986 <p>The COVID-19 outbreak resulted in the shutdown of athletic training facilities. Although the effects of these restrictions on daily activity and sleep patterns have been widely analyzed, the employed tools often lacked accuracy, and were based on subjective measures. This study assessed the effects of home confinement on objective physical activity (PA), physiological and sleep parameters in active individuals. Sixteen male elite fitness coaches (age: 29±3 years; height: 183±6 cm; body mass: 82±5 kg, body mass index: 24.7±1.8 kg/m<sup>2</sup>) participated in this retrospective study. One-way analysis of variance was conducted to analyze selected PA, physiological and sleep parameters collected by smartwatch (Garmin Fenix 6 pro, USA) data during four consecutive months [<em>i.e</em>., pre-confinement, 1<sup>st</sup> and 2<sup>nd</sup> months of confinement, and post-confinement, year 2020]. Ramadan intermittent fasting (RIF) month occurred during the 2<sup>nd</sup> month of confinement. Compared to pre-confinement, significant changes were registered for almost all parameters during the 1<sup>st</sup> and/or the 2<sup>nd</sup> month of confinements (p&lt;0.001), with <strong><em>(i)</em></strong> higher values for resting heart rate, sleep latency, and total, light and rapid eye movements sleep times (% change=7-523 %), and <strong><em>(ii)</em></strong> lower values for PA parameters, calories/day spent, average and highest respiratory rates, and deep sleep time during the home confinement period (% change=5-36 %). During the post-confinement month, all parameters regained pre-confinement values. In conclusion, home confinement-induced detraining negatively influenced the objective measurements of cardiorespiratory and sleep parameters among fitness coaches with a deeper effect during the 2<sup>nd</sup> month of home confinement, possibly due to the effect of RIF.</p> Ismail Dergaa Achraf Ammar Amine Souissi Mohamed Saifeddin Fessi Khaled Trabelsi Jordan M. Glenn Amine Ghram Morteza Taheri Khadijeh Irandoust Hamdi Chtourou Mohamed Romdhani Helmi Ben Saad Karim Chamari Copyright (c) 2022 Ismail Dergaa, Achraf Ammar, Amine Souissi, Mohamed Saifeddin Fessi, Khaled Trabelsi, Jordan M. Glenn, Amine Ghram, Morteza Taheri, Khadijeh Irandoust, Hamdi Chtourou, Mohamed Romdhani, Helmi Ben Saad, Karim Chamari http://creativecommons.org/licenses/by/4.0 2022-08-16 2022-08-16 21 1084 1098 10.17179/excli2022-4986 Okadaic acid influences xenobiotic metabolism in HepaRG cells https://www.excli.de/index.php/excli/article/view/5033 <p class="Abstract"><span lang="EN-US">Okadaic acid (OA) is an algae-produced lipophilic marine biotoxin that accumulates in the fatty tissue of filter-feeding shellfish. Ingestion of contaminated shellfish leads to the diarrheic shellfish poisoning syndrome. Furthermore, several other effects of OA like genotoxicity, liver toxicity and tumor-promoting properties have been observed, probably linked to the phosphatase-inhibiting properties of the toxin. It has been shown that at high doses OA can disrupt the physical barrier of the intestinal epithelium. As the intestine and the liver do not only constitute a physical, but also a metabolic barrier against xenobiotic exposure, we here investigated the impact of OA on the expression of cytochrome P450 (CYP) enzymes and transporter proteins in human HepaRG cells liver cells <em>in vitro </em>at non-cytotoxic concentrations. The interplay of OA with known CYP inducers was also studied. Data show that the expression of various xenobiotic-metabolizing CYPs was downregulated after exposure to OA. Moreover, OA was able to counteract the activation of CYPs by their inducers. A number of transporters were also mainly downregulated. Overall, we demonstrate that OA has a significant effect on xenobiotic metabolism barrier in liver cells, highlighting the possibility for interactions of OA exposure with the metabolism of drugs and xenobiotics.</span></p> Leonie T. D. Wuerger Helen S. Hammer Ute Hofmann Felicia Kudiabor Holger Sieg Albert Braeuning Copyright (c) 2022 Leonie T. D. Wuerger, Helen S. Hammer, Ute Hofmann, Felicia Kudiabor, Holger Sieg, Albert Braeuning http://creativecommons.org/licenses/by/4.0 2022-08-01 2022-08-01 21 1053 1065 10.17179/excli2022-5033 Inorganic arsenic contamination and the health of children living near an inactive mining site: northern Thailand https://www.excli.de/index.php/excli/article/view/4922 <p>Arsenic toxicity is a global health problem affecting millions of people. Contamination is caused by arsenic from natural geological sources leaching into aquifers, contaminating drinking water, and may also be caused by mining and other industrial processes. Acute arsenic poisoning is associated with nausea, vomiting, abdominal pain, and severe diarrhea. Chronic arsenic toxicity results in multisystemic diseases leading to central nervous system (CNS) impairments such as cognitive or intellectual deficits in children. Over the past ten years, arsenic contamination has been reported in northern Thailand. The Ministry of Public Health; Thailand, Forensic Science Institute Thammasat University, and the Research Center to Promote Safety and Prevent Injuries in Children at the Ramathibodi Hospital compiled a report on the health impact of the population within a 10 kilometer radius around a mine tailing in the Phichit, Phitsanulok, and Phetchabun Provinces of Thailand. It showed that more than 30 % of children (aged 8–13 years) had higher than normal arsenic contamination levels based on the Agency for Toxic Substances and Disease Registry (ATSDR). After the publication of that report, the mine was temporarily closed in 2016. Based on this data, this research aimed to follow arsenic contamination after the mining operation had stopped operation for three years. The study showed that 4.5 % of school aged children had levels of inorganic arsenic in their urine, higher than the normal range (ATSDR), showing clearly that inorganic arsenic contamination is still above the normal range in children living near an inactive mining site<strong>. </strong>Therefore, monitoring heavy metal contamination in Thailand and the health effects on vulnerable children who live near mines during regular operation or after being temporarily suspended can prevent and mitigate possible health impacts.</p> Sarun Kunwittaya Nootchanart Ruksee Thirata Khamnong Athiwat Jiawiwatkul Nonthasruang Kleebpung Vasunun Chumchua Adisak Plitponkarnpim Chutikorn Nopparat Kannika Permpoonputtana Copyright (c) 2022 Sarun Kunwittaya, Nootchanart Ruksee, Thirata Khamnong, Athiwat Jiawiwatkul, Nonthasruang Kleebpung, Vasunun Chumchua, Adisak Plitponkarnpim, Chutikorn Nopparat, Kannika Permpoonputtana http://creativecommons.org/licenses/by/4.0 2022-07-26 2022-07-26 21 1007 1014 10.17179/excli2022-4922 Ameliorative effects of high intensity interval training and Lactobacillus rhamnosus GG against tetracycline-induced fatty liver in rats https://www.excli.de/index.php/excli/article/view/4791 <p class="Abstract"><span lang="EN-US">Exercise training and probiotics have been suggested as a treatment for the prevention of chronic liver damage such as non-alcoholic fatty liver disease (NAFLD). <em>Lactobacillus rhamnosus Gorbach‐Goldin (LGG)</em> is one of the most widely used probiotic strains that decreases liver damage. Thus, this study aims to consider the ameliorative effects of high intensity interval training (HIIT) and <em>LGG</em> against tetracycline-induced fatty liver in rats. Eighty male Wistar rats were randomly divided into 8 groups of (n=10 each group): control, LGG, HIIT, LGG+HIIT, NAFLD, NAFLD+LGG, NAFLD+HIIT, and NAFLD+LGG+HIIT. The rats are treated by intraperitoneal injection with 140 mg/kg<sup>−1</sup> tetracycline, an antibiotic previously known to induce steatosis. The exercise training groups performed HIIT 5 days/week for 5 weeks. 10<sup>7</sup> colony-forming units (cfu) of LGG were gavaged for LGG groups 5 days/week for 5 weeks. Probiotic supplementation in combination with interval training significantly decreased <em>tissue inhibitor of matrix metalloproteinases-1 (TIMP-1)</em> mRNA and <em>matrix metalloproteinase-2</em> <em>(MMP-2)</em> mRNA in the liver (p&lt;0.05), while the levels of <em>lysosomal acid lipase</em> <em>(LIPA)</em> mRNA was significantly increased compared to NAFLD group. Also, compared with NAFLD group, NAFLD+LGG, NAFLD+HIIT and NAFLD+LGG+HIIT groups showed a significant decrease in hepatic <em>monocyte chemoattractant protein-1</em> <em>(MCP-1)</em>. Compared to LGG and LGG+HIIT groups, all NAFLD groups showed a significant decrease in <em>apolipoprotein C3 (APOC3)</em> in liver tissue (p&lt;0.05). The results suggested that interval exercise with LGG supplementation minimizes cell destruction and inflammation in liver tissue due to NAFLD by improving gene expression profiles.</span></p> Hamideh Mahmoodzadeh Hosseini Hossein Shirvani Fariba Aghaei Ehsan Arabzadeh Martin Hofmeister Copyright (c) 2022 Hamideh Mahmoodzadeh Hosseini, Hossein Shirvani, Fariba Aghaei, Ehsan Arabzadeh, Martin Hofmeister http://creativecommons.org/licenses/by/4.0 2022-07-21 2022-07-21 21 991 1006 10.17179/excli2022-4791 Tryptophan recovery index as a new biomarker for fitness https://www.excli.de/index.php/excli/article/view/4889 <p class="Abstract"><span lang="EN-US">The maximal oxygen uptake (VO<sub>2</sub>max) and maximal power output (P<sub>max</sub>) are commonly used parameters to evaluate the endurance fitness status. A connection between exercise and the kynurenine pathway (KP), which describes the metabolism of unused tryptophan, has already been reported. However, a potential association of the KP with endurance fitness levels remains unknown. In this study, adolescent competitive athletes performed an exhaustive incremental exercise test. Blood samples were taken before, directly after, and 30 minutes after the end of exercise. Tryptophan (Trp), kynurenine (Kyn) and kynurenic acid (KA) serum levels were determined by high-performance liquid chromatography (HPLC). Forty-four male and 27 female athletes (median age: 16 years) were recruited. During exhaustive exercise tests, Trp initially declined and then increased 30 minutes after discontinuing exercise. Similar findings were observed for Kyn, whereas KA levels behaved inversely. After incremental exhaustive exercise the relative increase of Trp concentrations, termed the tryptophan-recovery-index (TRI), showed a highly significant positive correlation with VO<sub>2</sub>max and P<sub>max </sub>(r=0.468 and 0.491, p-values &lt;0.001). There was a significant gender-difference with higher levels of all metabolites at all measured time points in male participants. In the present study, a highly significant correlation was found between the TRI and the maximal oxygen uptake in well-trained athletes. The implementation of TRI can therefore be suggested as a biomarker for physical fitness. </span></p> Alexander Pichler Andreas Meinitzer Dietmar Enko Peter Schober Georg Singer Christoph Castellani Markus Herrmann Sandra J. Holasek Holger Till Jana Maria Windhaber Copyright (c) 2022 Alexander Pichler, Andreas Meinitzer, Dietmar Enko, Peter Schober, Georg Singer, Christoph Castellani, Markus Herrmann, Sandra J. Holasek, Holger Till, Jana Maria Windhaber http://creativecommons.org/licenses/by/4.0 2022-06-24 2022-06-24 21 888 896 10.17179/excli2022-4889 Transient receptor potential channel involvement in antinociceptive effect of citral in orofacial acute and chronic pain models https://www.excli.de/index.php/excli/article/view/5042 <p class="Abstract"><span lang="EN-US">This study aimed to test for the possible antinociceptive effect of the naturally occurring terpene citral in rodent models of acute and chronic orofacial pain and to test for the possible involvement of transient receptor potential (TRP) channels in this effect. Acute nociceptive behavior was induced in one series of experiments by administering formalin, cinnamaldehyde, menthol or capsaicin to the upper lip. Nociceptive behavior was assessed by orofacial rubbing, and the effects of pre-treatment with citral (0.1, 0.3 or 1.0 mg/Kg) or vehicle (control) were tested on the behavior. Nociceptive behavior was also induced by formalin injected into the temporomandibular joint or mustard oil injected into the masseter muscle, preceded by citral or vehicle (control) treatment. The chronic pain model involved infraorbital nerve transection (IONX) that induced mechanical hypersensitivity which was assessed by von Frey hair stimulation of the upper lip. Motor activity was also evaluated. Docking experiments were performed using TRPV1 and TRPM8 channels. Citral but not vehicle produced significant (p&lt;0.01, ANOVA) antinociception on all the acute nociceptive behaviors, and these effects were attenuated by TRPV1 antagonist capsazepine, TRPM3 antagonist mefenamic acid and by TRPM8 desensitization, but not by ruthenium red and TRPA1 antagonist HC-030031. The IONX animals developed facial mechanical hypersensitivity that was significantly reduced by citral but not by vehicle. The docking experiments revealed that citral may interact with TRPV1 and TRPM8 channels. These results indicate the potential use of citral as an inhibitor of orofacial nociception in both acute and chronic pain states through TRPV1, TRPM3 and TRPM8 channels.</span></p> Sacha Aubrey Alves Rodrigues Santos Marina de Barros Mamede Vidal Damasceno Francisco Ernani Alves Magalhães Barry John Sessle Breytiner Amaro de Oliveira Francisco Lucas Alves Batista Antônio Eufrásio Vieira-Neto Adriana Rolim Campos Copyright (c) 2022 Sacha Aubrey Alves Rodrigues Santos, Marina de Barros Mamede Vidal Damasceno, Francisco Ernani Alves Magalhães, Barry John Sessle, Breytiner Amaro de Oliveira, Francisco Lucas Alves Batista, Antônio Eufrásio Vieira-Neto, Adriana Rolim Campos http://creativecommons.org/licenses/by/4.0 2022-06-24 2022-06-24 21 869 887 10.17179/excli2022-5042 Integrated bioinformatics analysis reveals that EZH2-rich domains promote transcriptional repression in cervical cancer https://www.excli.de/index.php/excli/article/view/5029 <p>Cervical cancer is the third female cancer most common worldwide. The carcinogenic process involves an alteration of the mechanisms associated with transcription. Several studies have reported an oncogenic role of the polycomb complex subunit, EZH2. However, the role of EZH2 in cervical cancer is unknown. Hence, the objective of this study was to determine the role of EZH2 in transcriptional regulation in cervical cancer. The EZH2 expression and the methylation status of its promoter were analyzed in The Cancer Genome Atlas. The EZH2 enrichment profile was analyzed using chromatin immunoprecipitation with massively parallel DNA sequencing data provided by ENCODE project. The chromatin compartments were identified in the 4D Nucleome Data Portal. The functional annotation was examined in Enrichr. We report that EZH2 expression is increased in cervical cancer which is associated with hypomethylation of its promoter. EZH2 is enriched at promoter and distal intergenic regions. We identified that EZH2 defines chromatin domains enriched with H3K27me3 within repressive compartments in the HeLa-S3 cell line. Additionally, high EZH2 expression is associated with the repression of the senescent phenotype in cervical cancer patients. Our results suggest the participation of EZH2 in the generation of domains with a silencer function in cervical cancer, which regulate the expression of genes associated with cellular senescence.</p> Eric Genaro Salmerón-Bárcenas Ana Elvira Zacapala-Gómez Julio Ortiz-Ortiz Francisco Israel Torres-Rojas Pedro Antonio Avila-Lopez Copyright (c) 2022 Eric Genaro Salmerón-Bárcenas, Ana Elvira Zacapala-Gómez, Julio Ortiz-Ortiz, Francisco Israel Torres-Rojas, Pedro Antonio Avila-Lopez http://creativecommons.org/licenses/by/4.0 2022-06-23 2022-06-23 21 852 868 10.17179/excli2022-5029 Immunomodulatory components of Trichinella spiralis excretory-secretory products with lactose-binding specificity https://www.excli.de/index.php/excli/article/view/4954 <p class="Abstract"><span lang="EN-US">The immunomodulatory potential of <em>Trichinella spiralis</em> muscle larvae excretory-secretory products (ES L1) has been well documented <em>in vitro </em>on dendritic cells (DCs) and in animal models of autoimmune diseases. ES L1 products possess the potential to induce tolerogenic DCs and consequently trigger regulatory mechanisms that maintain immune homeostasis. The use of ES L1 as a potential treatment for various inflammatory disorders proved to be beneficial in animal models, although the precise immunomodulatory factors have not yet been identified. This study aimed at the isolation and characterization of ES L1 components that possess galectin family member properties. Galectin-1-like proteins (TsGal-1-like) were isolated from ES L1 based on the assumption of the existence of a lactose-specific carbohydrate-recognition domain and were recognized by anti-galectin-1 antibodies in Western blot. This TsGal-1-like isolate, similar to galectin-1, induced DCs with tolerogenic properties and hence, the capacity to polarize T cell response towards a regulatory type. This was reflected by a significantly increased percentage of CD4<sup>+</sup>CD25<sup>+</sup>Foxp3<sup>+</sup> regulatory T cells and significantly increased expression of IL-10 and TGF-β within this cell population. Proteomic analysis of TsGal-1-like isolate by mass spectrometry identified nineteen proteins, seven with annotated function after blast analysis against a database for <em>T. spiralis</em> and the UniProt database. To our surprise, none of the identified proteins possesses homology with known galectin family members. Nevertheless, the isolated components of ES L1 possess certain galectin-1 properties, such as specific lactose binding and the potential to elicit a regulatory immune response, so it would be worth further investigating the structure of sugar binding within isolated proteins and its biological significance.</span></p> Natasa Ilic Zanka Bojic-Trbojevic Britta Lundström-Stadelmann Danica Cujic Ivana Mitic Alisa Gruden-Movsesijan Copyright (c) 2022 Natasa Ilic, Zanka Bojic-Trbojevic, Britta Lundström-Stadelmann, Danica Cujic, Ivana Mitic, Alisa Gruden-Movsesijan http://creativecommons.org/licenses/by/4.0 2022-06-03 2022-06-03 21 793 813 10.17179/excli2022-4954 Plasma lncRNA profiling identified BC200 and NEAT1 lncRNAs as potential blood-based biomarkers for late-onset Alzheimer’s disease https://www.excli.de/index.php/excli/article/view/4764 <p>Long non-coding RNAs (lncRNA) play critical roles in pathogenesis of neurodegenerative diseases. Human plasma carries lncRNAs that are stable in the blood, and their disease-specific profile have made them valuable biomarkers for some diseases. This study reports screening of the plasma levels of 90 lncRNAs in patients with Alzheimer disease (AD) to find out plasma-based AD biomarkers. Total RNA was isolated from plasma samples of 50 AD and 50 matched healthy controls. The plasma samples of 10 advanced AD patients and 10 matched healthy controls were screened for expression levels of 90 lncRNAs using Human LncRNA Profiler qPCR Array Kit (SBI). Based on the profiling results, lncRNAs BC200, NDM29, NEAT1, FAS-AS1 and GAS5-AS1 were selected for further analysis in all samples and their biomarker potency was evaluated by ROC curve analysis. We further surveyed RNAseq data by <em>in silico</em> analysis. We found that the NEAT1 and BC200 levels in the plasma of the AD patients were significantly higher compared with the control group (P=0.0021, p= 0.02, respectively). ROC curve analysis showed that the plasma level of NEAT1 and BC200 discriminated AD patients from healthy controls with sensitivity of 72 % and 60 %, and specificity of 84 % and 91 % respectively. Moreover, NEAT1 discriminated MCI (60 % sensitivity and 91 % specificity) and advanced-AD patients from healthy controls (73 % sensitivity and 71 % specificity). Besides, plasma level of BC200 discriminated the pre-clinical subjects from healthy controls with 83 % sensitivity and 66 % specificity. A positive correlation was also observed between plasma levels of BC200 with the age patients (r = 0.34, p=0.02). In silico RNAseq data analysis showed that a total of 33 lncRNAs were up-regulated but 13 lncRNAs were down-regulated significantly in AD patients compared with the healthy controls. In conclusion, this study elucidated that the plasma levels of lncRNAs NEAT1 and BC200 might be considered as potential blood-based biomarkers for AD development and progression.</p> Majid Khodayi Mohammad Khalaj-Kondori Mohammad Ali Hoseinpour Feizi Mortaza Jabarpour Bonyadi Mahnaz Talebi Copyright (c) 2022 Majid Khodayi, Mohammad Khalaj-Kondori, Mohammad Ali Hoseinpour Feizi, Mortaza Jabarpour Bonyadi, Mahnaz Talebi http://creativecommons.org/licenses/by/4.0 2022-05-09 2022-05-09 21 772 785 10.17179/excli2022-4764 Ergonomic assessment of postal workers’ pain symptoms and musculoskeletal risks related to parcel processing activity for delivery https://www.excli.de/index.php/excli/article/view/4857 <p class="Abstract"><span lang="EN-US">The parcel delivery activity is carried out all over the world and workers in this sector have suffered from musculoskeletal disorders (MSDs) due to the strong demand for work generated by the recent increase in e-commerce. This study aimed to evaluate postal workers' pain symptoms, movements and identify MSDs risks related to the parcel processing activity for delivery, proposing preventive measures. A sample of thirty-two workers was evaluated with the application of sociodemographic and Nordic questionnaires and electrical bioimpedance. The motion capture sensors were used to evaluate right/left shoulder joints, segment C7-T1 (Cervical) and segment L5-S1 (Lumbar) of three postal workers (percentiles of anthropometric data: 5, 50, and 95) during four real work activities that are part of the parcel processing. The analyzed workers presented musculoskeletal complaints in practically all body regions, with a greater prevalence in shoulders, hands, lower back, and knees. According to the Body Mass Index (BMI), they were on average overweight (27.8 ± 3.7 kg/m<sup>2</sup>). In the movement analysis, we identified risks related to cervical protrusion, anterior trunk flexion, and shoulder flexion, in addition to repetitive movements. In some activities, the higher stature showed an increase in lumbar and cervical anterior flexion. The set of evaluations showed that the activity of processing orders for delivery offers musculoskeletal risks. We identify that ergonomic adaptations are necessary to adapt the heights of the work environment to the statures of the postal workers. Relevance to industry: The activity of processing orders for delivery is carried out practically all over the world generating jobs and income for its employees. Nonetheless, there are still situations of ergonomic disadvantage that can generate musculoskeletal risks. The findings elucidate ergonomic risks and provide useful information for future ergonomic interventions in the postal/delivery workplace environment.</span></p> Lincoln Silva Nélson Costa Carolina Schutz Rosa Irandir Izaquiel Paulo Natália Mattos da Silva Cesar Giracca Sabrina da Silveira Simões Adriana do Nascimento Aquini Giselle Merino Eugenio Andrés Díaz Merino Copyright (c) 2022 Lincoln Silva, Nélson Costa, Carolina Schutz Rosa, Irandir Izaquiel Paulo, Natália Mattos da Silva, Cesar Giracca, Sabrina da Silveira Simões, Adriana do Nascimento Aquini, Giselle Merino, Eugenio Andrés Díaz Merino http://creativecommons.org/licenses/by/4.0 2022-04-27 2022-04-27 21 744 756 10.17179/excli2022-4857 Cytotoxic capability and the associated proteomic profile of cell-free coelomic fluid extracts from the edible sea cucumber Holothuria tubulosa on HepG2 liver cancer cells https://www.excli.de/index.php/excli/article/view/4825 <p class="Abstract"><span lang="EN-US">Hepatocellular carcinoma (HCC) is an aggressive cancer histotype and one of the most common types of cancer worldwide. The identification of compounds that might intervene to restrain neoplastic cell growth appears imperative due to its elevated overall mortality. The marine environment represents a reservoir rich in bioactive compounds in terms of primary and secondary metabolites produced by aquatic animals, mainly invertebrates. In the present study, we determined whether the water-soluble cell-free extract of the coelomic fluid (CFE) of the edible sea cucumber <em>Holothuria tubulosa</em> could play an anti-HCC role <em>in vitro </em>by analyzing the viability and locomotory behavior, cell cycle distribution, apoptosis and autophagy modulation, mitochondrial function and cell redox state of HepG2 HCC cells. We showed that CFE causes an early block in the cell cycle at the G<sub>2</sub>/M phase, which is coupled to oxidative stress promotion, autophagosome depletion and mitochondrial dysfunction ultimately leading to apoptotic death. We also performed a proteomic analysis of CFE identifying a number of proteins that are seemingly responsible for anti-cancer effects. In conclusion, <em>H. tubulosa</em>’s CFE merits further investigation to develop novel promising anti-HCC prevention and/or treatment agents and also beneficial supplements for formulation of functional foods and food packaging material.</span></p> Claudio Luparello Rossella Branni Giulia Abruscato Valentina Lazzara Laszlo Drahos Vincenzo Arizza Manuela Mauro Vita Di Stefano Mirella Vazzana Copyright (c) 2022 Claudio Luparello, Rossella Branni, Giulia Abruscato, Valentina Lazzara, Laszlo Drahos, Vincenzo Arizza, Manuela Mauro, Vita Di Stefano, Mirella Vazzana http://creativecommons.org/licenses/by/4.0 2022-04-25 2022-04-25 21 722 743 10.17179/excli2022-4825 Transplantation of SDF-1α-loaded liver extracellular matrix repopulated with autologous cells attenuated liver fibrosis in a rat model https://www.excli.de/index.php/excli/article/view/4761 <p>Cell-based therapy and tissue engineering are promising substitutes for liver transplantation to cure end-stage liver disorders. However, the limited sources for healthy and functional cells and poor engraftment rate are main challenges to the cell-based therapy approach. On the other hand, feasibility of production and size of bioengineered tissues are primary bottlenecks in tissue engineering. Here, we induce regeneration in a rat fibrotic liver model by transplanting a natural bioengineered scaffold with a native microenvironment repopulated with autologous stem/progenitor cells. In the main experimental group, a 1 mm<sup>3</sup> stromal derived factor-1α (SDF-1α; S) loaded scaffold from decellularized liver extracellular matrix (LEM) was transplanted (Tx) into a fibrotic liver and the endogenous stem/progenitor cells were mobilized via granulocyte colony stimulating factor (G-CSF; G) therapy. Four weeks after transplantation, changes in liver fibrosis and necrosis, efficacy of cell engraftment and differentiation, vasculogenesis, and liver function recovery were assessed in this (LEM-TxSG) group and compared to the other groups. We found significant reduction in liver fibrosis stage in the LEM-TxSG, LEM-TxS and LEM-TxG groups compared to the control (fibrotic) group. Liver necrosis grade, and alanine transaminase (ALT) and aspartate transaminase (AST) levels dramatically reduced in all experimental groups compared to the control group. However, the number of engrafted cells into the transplanted scaffold and ratio of albumin (Alb) positive cells per total incorporated cells were considerably higher in the LEM-TxSG group compared to the LEM-Tx, LEM-TxS and LEM-TxG groups. Serum Alb levels increased in the LEM-Tx, LEM-TxS, and LEM-TxG groups, and was highest in the LEM-TxSG group, which was significantly more than the fibrotic group. Small vessel formation in the LEM-TxSG group was significantly higher than the LEM-Tx and LEM-TxS groups. Totally, these findings support application of the <em>in vivo</em> tissue engineering approach as a possible novel therapeutic strategy for liver fibrosis.</p> Mostafa Najar-Asl Hossein Bahadoran Mohammad-Hossein Asadi Mona Saheli Mohammad-Hassan Asghari Niloofar Sodeifi Mohammad Kazemi Ashtiani Massoud Vosough Hossein Baharvand Abbas Piryaei Copyright (c) 2022 Mostafa Najar-Asl, Hossein Bahadoran, Mohammad-Hossein Asadi, Mona Saheli, Mohammad-Hassan Asghari, Niloofar Sodeifi, Mohammad Kazemi Ashtiani, Massoud Vosough, Hossein Baharvand, Abbas Piryaei http://creativecommons.org/licenses/by/4.0 2022-04-22 2022-04-22 21 704 721 10.17179/excli2022-4761 Medical history, medication use and physical activity in adults in their eighth and ninth decade of life in the Hertfordshire Cohort Study https://www.excli.de/index.php/excli/article/view/4874 <p>While there are many known health benefits to maintained physical activity levels in late adulthood, there have been very few studies that have considered relationships between morbidity profile and physical activity in the eighth decade of life. We studied 1097 participants, 555 men and 542 women from the Hertfordshire Cohort Study, a UK community based sample. Validated questionnaire based data were used to relate self-reported physical activity (PA) levels to medical history, and medication use. Regression analyses were adjusted for age, BMI, smoker status, alcohol consumption. The mean (SD) age of participants in the study was 80.2 (2.7) years for men and 80.2 (2.6) for women. A higher proportion of men (33.7 %) than women (24 %) were in the high activity score group. 20.8 % of female participants and 22.6 % male participants reported having no comorbid disease; 10.5 % men and 8.4 % women were taking no medication. Higher number of chronic conditions was associated with lower levels of PA [men (OR 0.73, 95 % CI 0.63-0.84, p&lt;0.001); women (OR 0.74, 95 % CI 0.64-0.86, p&lt;0.001)] as was being prescribed a higher number of medications [men (OR 0.88, 95 % CI 0.84-0.93, p&lt;0.001); women (OR 0.86, 95 % CI 0.82-0.91, p&lt;0.001)]. All these associations remained robust following adjustments. Strong relationships were seen in both sexes between PA and taking medication for disorders of the central nervous system and gastrointestinal system, with relationships generally stronger in men. We have observed relationships between comorbid medical history and medication use with physical activity in a cohort of community dwelling older adults. These highlight the need to consider medical history when considering how best to optimize PA in older adults.</p> Gregorio Bevilacqua Jean Zhang Camille Parsons Faidra Laskou Nicholas Fuggle Cyrus Cooper Elaine Dennison Copyright (c) 2022 Gregorio Bevilacqua, Jean Zhang, Camille Parsons, Faidra Laskou, Nicholas Fuggle, Cyrus Cooper, Elaine Dennison http://creativecommons.org/licenses/by/4.0 2022-04-19 2022-04-19 21 695 703 10.17179/excli2022-4874 The “identikit” of subject with obesity and COVID-19 vaccine breakthrough https://www.excli.de/index.php/excli/article/view/4864 <p class="Abstract"><span lang="EN-US">The mRNA coronavirus disease 2019 (COVID-19) vaccines were highly effective in the prevention of symptomatic COVID-19, hospitalization, severe disease, and death. However, a minority of vaccinated individuals might become infected and experience significant morbidity. Risk factors of COVID-19 vaccine breakthrough in obesity have not been elucidated. Thus, we aimed to portray the subjects with obesity developing COVID-19 vaccine breakthrough despite vaccination. Coronavirus 2019 (COVID-19) mRNA vaccines have been highly effective in preventing symptomatic COVID-19, hospitalization, severe illness and death. However, a minority of vaccinated individuals may become infected and experience considerable morbidity. The risk factors for COVID-19 vaccine breakthrough in obesity have not been elucidated. Therefore, we aimed to depict individuals with obesity who develop COVID-19 vaccine breakthrough despite vaccination. An online questionnaire was distributed to respondents via a snowball sampling method among subjects with obesity belonging to Italian Associations for people living with obesity aged 18 years and above. Two hundred and thirty-five respondents (44.5±14 years; BMI: 33.3±7.2 kg/m<sup>2</sup>) were included in the study. COVID-19 vaccine breakthrough was noted in 34 % of respondents. A higher prevalence of grade III obesity was detected in subjects with COVID-19 vaccine breakthrough compared to subjects that did not (27.5 % <em>vs</em> 13.5 %; p=0.014). In addition, a significant lower prevalence of respondents that completed third dose were found in respondents with COVID-19 vaccine breakthrough compared with respondents that did not develop it (33.8 % <em>vs </em>72.9 %; p&lt;0.001). After stratifying respondents with COVID-19 vaccine breakthrough according to the completed doses of vaccine, we found that, although no differences were detected in terms of clinical manifestations of COVID-19, there was a significant higher prevalence of type 2 diabetes and hypertension in respondents that completed third doses compared to respondents that completed first and second doses. In conclusion, COVID-19 vaccine breakthrough was more common in subjects with grade III obesity. The presence of type 2 diabetes and hypertension could counteract the immune potentiating effects of vaccine booster against COVID-19.</span></p> Giovanna Muscogiuri Luigi Barrea Ludovica Verde Claudia Vetrani Silvia Savastano Annamaria Colao Copyright (c) 2022 Giovanna Muscogiuri, Luigi Barrea, Ludovica Verde, Claudia Vetrani, Silvia Savastano, Annamaria Colao http://creativecommons.org/licenses/by/4.0 2022-04-08 2022-04-08 21 687 694 10.17179/excli2022-4864 The diaryl-imidazopyridazine anti-plasmodial compound, MMV652103, exhibits anti-breast cancer activity https://www.excli.de/index.php/excli/article/view/4323 <p class="Abstract"><span lang="EN-US">Breast cancer is the most common malignancy in women worldwide and it remains a global health burden, in part, due to poor response and tolerance to current therapeutics. Drug r<span style="color: black;">epurposing, which seeks to </span>identify new indications for existing and investigational drugs,<span style="color: black;"> has become an exciting strategy to address these challenges</span>. Here we describe the anti-breast cancer activity of a <span style="color: black;">diaryl-imidazopyridazine compound, <a name="_Hlk81473567"></a>MMV652103, which was previously identified for its anti-plasmodial activity. We demonstrate that MMV652103 potently inhibits the oncogenic PI4KB and PIK3C2G lipid kinases, is selectively cytotoxic to MCF7 and T47D estrogen receptor positive breast cancer cells and inhibits their ability to survive and migrate. The underlying mechanisms involved included the induction of reactive oxygen species and activation of the DNA damage and p38 MAPK stress signaling pathways. This was associated with a G1 cell cycle arrest and an increase in levels of the cyclin-dependent kinase inhibitor p21 and activation of apoptotic and autophagic cell death pathways. Lastly, MMV652103 significantly reduced the weight and metastases of MCF7 induced tumors in an <em>in vivo</em> chick embryo model and displayed a favorable safety profile. These findings position MMV652103 as a promising chemotherapeutic in the treatment of oestrogen receptor positive breast cancers.</span></span></p> Alexis Neumann-Mufweba Serah Kimani Saif Feroz Khan Kelly Chibale Sharon Prince Copyright (c) 2022 Alexis Neumann-Mufweba, Serah Kimani, Saif Feroz Khan, Kelly Chibale, Sharon Prince http://creativecommons.org/licenses/by/4.0 2022-04-04 2022-04-04 21 656 679 10.17179/excli2021-4323 Analyzing the interaction of human ACE2 and RBD of spike protein of SARS-CoV-2 in perspective of Omicron variant https://www.excli.de/index.php/excli/article/view/4721 <p class="Abstract"><span lang="EN-US">The newly identified Omicron (B.1.1.529) variant of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) has steered concerns across the world due to the possession of a large number of mutations leading to high infectivity and vaccine escape potential. The Omicron variant houses 32 mutations in spike (S) protein alone. The viral infectivity is determined mainly by the ability of S protein Receptor Binding Domain (RBD) to bind to the human Angiotensin I Converting Enzyme 2 (hACE2) receptor. In this paper, the interaction of the RBDs of SARS-CoV-2 variants with hACE2 was analyzed by using protein-protein docking and compared with the novel Omicron variant. Our findings reveal that the Omicron RBD interacts strongly with hACE2 receptor via unique amino acid residues as compared to the Wuhan and many other variants. However, the interacting residues of RBD are found to be the same in Lamda (C.37) variant. This unique binding of Omicron RBD with hACE2 suggests an increased potential of infectivity and vaccine evasion potential of the new variant. The evolutionary drive of the SARS-CoV-2 may not be exclusively driven by RBD variants but surely provides for the platform for emergence of new variants.</span></p> Arijit Samanta Syed Sahajada Mahafujul Alam Safdar Ali Mehboob Hoque Copyright (c) 2022 Arijit Samanta, Syed Sahajada Mahafujul Alam, Safdar Ali, Mehboob Hoque http://creativecommons.org/licenses/by/4.0 2022-03-10 2022-03-10 21 610 620 10.17179/excli2022-4721 Low-toxic herbicides Roundup and Atrazine disturb free radical processes in Daphnia in environmentally relevant concentrations https://www.excli.de/index.php/excli/article/view/4690 <p class="Abstract"><span lang="EN-US">The use of glyphosate-based Roundup and triazine herbicide Atrazine has increased markedly in last decades. Thus, it is important to evaluate toxic effects of these herbicides to non-targeted organisms such as zooplankton to understand their safety toward aquatic ecosystems. In the current study, we performed Daphnia toxicity tests based on lethality to identify LC<sub>50</sub> that provides acute aquatic toxicity classification criteria. LC<sub>50</sub> for Roundup exposure for 24 hours was found to be 0.022 mg/L and 48 hours - 0.0008 mg/L. Atrazine showed LC<sub>50</sub> at concentrations of 40 mg/L and 7 mg/L for 24 and 48 hours, respectively. We demonstrated that exposure to ecologically relevant concentrations of Roundup or Atrazine decreases lipid peroxidation and protein thiol levels, however caused increase in carbonyl protein and low-molecular-weight thiols content. Moreover, the herbicide treatments caused increase of superoxide dismutase activity. Our data suggest that at very low concentrations Roundup and Atrazine disturb free radical processes in <em>D. magna</em>.</span></p> Viktor Husak Tetiana Strutynska Nadia Burdyliuk Anzhelika Pitukh Volodymyr Bubalo Halyna Falsfushynska Olha Strilbytska Oleh Lushchak Copyright (c) 2022 Viktor Husak, Tetiana Strutynska, Nadia Burdyliuk, Anzhelika Pitukh, Volodymyr Bubalo, Halyna Falsfushynska, Olha Strilbytska, Oleh Lushchak http://creativecommons.org/licenses/by/4.0 2022-03-09 2022-03-09 21 595 609 10.17179/excli2022-4690 Analysis of hematological parameters in rheumatoid arthritis patients receiving biological therapy https://www.excli.de/index.php/excli/article/view/4702 <p class="Abstract"><span lang="EN-US">Administration of biological therapy (BT) in rheumatoid arthritis (RA) patients is often associated with hematological complications, which result in switching among therapies. Thus, there is an instant need for suitable screening parameters that will help to individualize the therapy and minimize the onset of adverse effects. We analyzed the hematological profile of 99 RA patients receiving TNFα (Adalimumab - ADA, Golimumab - GOL, Etanercept - ETA) or IL-6 receptor (Tocilizumab - TCZ) inhibitors in order to find possible indicators to improve personalization of RA therapy. BTs significantly affect the levels of observed hematological parameters. In contrast to TNF-α inhibitors, TCZ normalized almost all monitored hematological parameters to values of healthy donors. Only GOL from the TNF-α inhibitors studied, was able to normalize neutrophil counts, as well as platelet indicators. Importantly, effects on the blood parameters (e.g. lymphocytes or platelet count) differ even within the same therapeutic group (anti-TNFα). Variable effects of individual biological agents in RA treatment point to importance to evaluate the patient’s hematological profile to improve the selection of suitable BT. It will help to personalize the administration of BT and prevent unnecessary switching from an effective therapy just because of provocation of avoidable hematological complications.</span></p> Jana Pereckova Silvia Martiniakova Juraj Payer Martin Falk Zdenko Killinger Tomas Perecko Copyright (c) 2022 Jana Pereckova, Silvia Martiniakova, Juraj Payer, Martin Falk, Zdenko Killinger, Tomas Perecko http://creativecommons.org/licenses/by/4.0 2022-03-08 2022-03-08 21 580 594 10.17179/excli2022-4702 Hypoxia-inducible factor (HIF)-3α2 serves as an endothelial cell fate executor during chronic hypoxia https://www.excli.de/index.php/excli/article/view/4622 <p>The adaptive response to hypoxia involves the transcriptional induction of three transcription factors called hypoxia inducible factor alpha 1, 2 and 3 (HIF-1α, HIF-2α, and HIF-3α) which dimerize with constitutively expressed beta chains that together form the HIF-1, -2 and -3 transcription factors. During normoxic conditions, the alpha chain is expressed at low levels since its stability is regulated by prolyl-hydroxylation that promotes subsequent ubiquitination and degradation. During hypoxic conditions, however, the prolyl hydroxylases are less active, and the alpha chain accumulates through elevated protein stability and the elevated induction of expression. Two of the three HIFs isoforms present in mammals, HIF-1 and HIF-2, are well characterized and have overlapping functions that promote cell survival, whereas HIF-3’s role remains less clear. The HIF-3 response is complicated because the <em>HIF3A</em> gene can utilize different promotors and alternate splicing sites that result in a number of different HIF-3α isoforms. Here, using human umbilical vein endothelial cells (HUVECs), we demonstrate that one of the isoforms of HIF-3α, isoform 2 (HIF-3α2) accumulates at a late stage of hypoxia and induces the expression of DNA damage inducible transcript 3 (<em>DDIT4</em>), a gene known to promote apoptosis. We also demonstrate that caspase 3/7 activity is elevated, supporting that the role of the HIF-3α2 isoform is to promote apoptosis. Furthermore, we provide evidence that HIF-3α2 is also expressed in seven other primary endothelial cell types, suggesting that this may be a common feature of HIF-3α2 in endothelial cells. </p> Maciej Jaśkiewicz Adrianna Moszyńska Marcin Serocki Jarosław Króliczewski Sylwia Bartoszewska James F. Collawn Rafal Bartoszewski Copyright (c) 2022 Maciej Jaśkiewicz, Adrianna Moszyńska, Marcin Serocki, Jarosław Króliczewski, Sylwia Bartoszewska, James F. Collawn, Rafal Bartoszewski http://creativecommons.org/licenses/by/4.0 2022-02-21 2022-02-21 21 454 469 10.17179/excli2021-4622 IKBKE-driven TPL2 and MEK1 phosphorylations sustain constitutive ERK1/2 activation in tumor cells https://www.excli.de/index.php/excli/article/view/4578 <p class="Abstract"><span lang="EN-US">IKBKE have been associated with numerous cancers. As a result, IKBKE have emerged as potential target for cancer therapy. Accumulating evidence support that IKBKE orchestrate tumor cell survival in cancers. Here we evaluated the possible link between IKBKE and ERK phosphorylation. The effects of IKBKE silencing on MAPK activation in tumor vs. normal cells were evaluated via WB and RT-PCR. Ectopically expressed IKBKE, TPL2 or MEK1 constructs were used to examine the possible interactions among them via co-IP. <em>In vitro</em> kinase assays were performed to understand nature of the observed interactions. In tumors, IKBKE regulates MEK/ERK constitutive activations <em>in vitro </em>and<em> in vivo</em>. IKBKE and TPL2 physically interact and this interaction leads to TPL2 phosphorylation. We describe here a novel regulatory link between IKBKE and constitutive ERK1/2 activation in tumor cells. This new circuitry may be relevant for tumor cell survival in various malignancies.</span></p> Serkan Ismail Göktuna Copyright (c) 2022 Serkan İsmail Göktuna http://creativecommons.org/licenses/by/4.0 2022-02-18 2022-02-18 21 436 453 10.17179/excli2021-4578 Helicobacter pylori infection and lactose intolerance increase expiratory hydrogen https://www.excli.de/index.php/excli/article/view/4508 <p>Infection with <em>Helicobacter pylori</em> (<em>H.pylori</em>) may cause dyspepsia and/or unexplained functional nonspecific, gastrointestinal complaints of the irritable bowel syndrome (IBS) spectrum. Hitherto, in <em>H. pylori</em> infected patients with symptoms of the IBS spectrum the occurrence of additional food intolerance/malabsorption is not evaluated. We used a retrospective analysis of charts from 548 patients who presented with gastrointestinal complaints of the irritable bowel syndrome spectrum. An enzyme-linked IgA immunosorbent assay or histologic evaluation of gastric mucosa were used to detect <em>H. pylori </em>infection. A hydrogen breath (H<sub>2</sub>) test was performed to evaluate fructose malabsorption (FM) and lactose intolerance (LIT). Serum diamine oxidase value of &lt;10 U/ml and a response to a histamine-reduced diet was used to identify histamine intolerance (HIT). We found 293 patients infected with<em> H. pylori</em>, within these were 58 <em>H. pylori </em>patients with LIT, 23 <em>H. pylori </em>LIT patients with FM and 46 <em>H. pylori </em>LIT patients with HIT. Additionally, 13 <em>H. pylori,</em> lactose- and histamine intolerance patients also had FM. The Kruskal Wallis test and pairwise comparison were used to analyze differences of the area under the curve of expiratory hydrogen. In lactose H<sub>2</sub> breath tests compared with LIT-only patients, LIT with <em>H. pylori</em>, LIT and <em>H. pylori</em> with HIT, LIT and <em>H. pylori</em> with FM showed significantly higher exhaled H<sub>2</sub> levels (p=0.022). Pairwise comparison demonstrated <em>H. pylori</em> infected patients with LIT exhaled more H<sub>2</sub> compared to LIT-only (p=0.029). <em>H. pylori </em>with lactose- and histamine intolerance, and <em>H. pylori </em>with lactose-, histamine intolerance and FM compared to <em>H. pylori</em>-only patients indicated a significantly higher occurrence of stomach pain during lactose H<sub>2</sub> breath tests (p=0.012 and p=0.005, respectively). We demonstrate that LIT patients with high expiratory H<sub>2</sub> levels in lactose breath tests may have <em>H. pylori</em> infection and possibly additional food intolerance/malabsorption. Subsequently, besides <em>H. pylori </em>eradication, a dietician is necessary for an individually tailored reduction- or exclusion diet of symptom triggering food components.</p> Wolfgang J. Schnedl Nathalie Meier-Allard Michael Schenk Sonja Lackner Dietmar Enko Harald Mangge Sandra J. Holasek Copyright (c) 2022 Wolfgang J. Schnedl, Nathalie Meier-Allard, Michael Schenk, Sonja Lackner, Dietmar Enko, Harald Mangge, Sandra J. Holasek http://creativecommons.org/licenses/by/4.0 2022-02-17 2022-02-17 21 426 435 10.17179/excli2021-4508 Molecular docking and mouse modeling suggest CMKLR1 and INSR as targets for improving PCOS phenotypes by minocycline https://www.excli.de/index.php/excli/article/view/4534 <p class="Abstract"><span lang="EN-US">Polycystic ovary syndrome (PCOS) is the most common cause of women’s infertility. Some inflammatory pathways play a pivotal role in the pathogenesis of PCOS. This study aimed to investigate the possible beneficial effects of minocycline on chemokine-like receptor 1 (CMKLR1) and Insulin Receptor (INSR) in a PCOS model. A molecular docking study was implemented using Molecular Operating Environment (MOE) software. The PCOS was induced in NMRI mice (mean body weight 14.47±0.23) by 28 days estradiol valerate injection (2 mg/kg/day). The mice were then divided into six groups (n=8 per group, mean body weight 17.77± 0.26): control (received normal saline), PCOS model, control for minocycline, minocycline treated PCOS (50 mg/kg), letrozole treated PCOS (0.5 mg/kg), and metformin-treated PCOS (300 mg/kg). Serum FSH, LH, estradiol (E2), and testosterone were detected by ELISA. The ovarian tissues were stained by hematoxylin and eosin. The CMKLR1 and INSR expression levels were determined by Real-time-PCR. The molecular docking studies showed scores of -10.92 and -9.30 kcal/mol, respectively, for minocycline with CMKLR1 and INSR. Estradiol valerate treatment led to a significant increase in E2, graffian follicle, and decrease in corpus luteum (CL) numbers (P&lt;0.05), while minocycline treatment improved these PCOS features. The minocycline treatment significantly decreased the CMKLR1 expression and increased the INSR expression (P&lt;0.05) while the CMKLR1 expression was increased in PCOS model. Minocycline may improve ovulation in PCOS model by returning E2 to a normal level and increasing CL number (ovulation signs). These beneficial outcomes may be related to the changes in CMKLR1 and INSR gene expression involved in glucose metabolism and inflammation.</span></p> Mahdie Kian Elham Hosseini Tooba Abdizadeh Taimour Langaee Azadeh Khajouei Sorayya Ghasemi Copyright (c) 2022 Mahdie Kian, Elham Hosseini, Tooba Abdizadeh, Taimour Langaee, Azadeh Khajouei, Sorayya Ghasemi http://creativecommons.org/licenses/by/4.0 2022-02-16 2022-02-16 21 400 414 10.17179/excli2021-4534 Phlomis fruticosa L. exerts in vitro antineurodegenerative and antioxidant activities and induces prooxidant effect in glioblastoma cell line https://www.excli.de/index.php/excli/article/view/4487 <p>Despite the significant advances in drug development we are witnessing the inability of health systems to combat both neurodegenerative diseases and cancers, especially glioblastoma. Hence, natural products are comprehensively studied in order to provide novel therapeutic options. This study aimed to explore anti-neurodegenerative and anti-glioblastoma potential of extract of <em>Phlomis fruticosa </em>L. using <em>in vitro</em> model systems. It was found that the methanol extract of <em>P. fruticosa</em> was able to efficiently reduce activities of enzymes linked to neurodegenerative disease including acetylcholinesterase, butyrylcholinesterase and tyrosinase. Furthermore, <em>P. fruticosa</em> extract has shown excellent antioxidant potential, as evidenced by six different methods. Analysis of cytotoxic effect of <em>P. fruticosa</em> extract on A172 glioblastoma cell line revealed that the concentration of the extract necessary for 50 % inhibition of A172 growth (IC<sub>50</sub>) was 710 μg/mL. The extract did not induce changes in proliferation and morphology of A172 glioblastoma cells. On the other side, production of ROS was increased in A172 cells treated with the extract. Observed cytotoxic effect of <em>P. fruticosa</em> extract might be based on increase in ROS generation upon treatment. Quantitative chemical analysis revealed the presence of twelve different polyphenols with the <em>cis</em> 3-<em>O</em>-caffeoylquinic acid being the most abundant. This study provided scientific evidence for further exploration of <em>P. fruticosa</em> as a promising natural anti-neurodegenerative therapeutic option.</p> Dejan Stojković Danijela Drakulić Maria Inês Dias Gokhan Zengin Lillian Barros Marija Ivanov Uroš Gašić Nemanja Rajčević Milena Stevanović Isabel C. F. R. Ferreira Marina Soković Copyright (c) 2022 Dejan Stojković, Danijela Drakulić, Maria Inês Dias, Gokhan Zengin, Lillian Barros, Marija Ivanov, Uroš Gašić, Nemanja Rajčević, Milena Stevanović, Isabel C. F. R. Ferreira, Marina Soković http://creativecommons.org/licenses/by/4.0 2022-02-14 2022-02-14 21 387 399 10.17179/excli2021-4487 Synthesis of acetamidosulfonamide derivatives with antioxidative and QSAR studies https://www.excli.de/index.php/excli/article/view/4590 <p>A series of sixteen acetamidosulfonamide derivatives (<strong>1</strong>-<strong>16</strong>) have been synthesized and investigated for their antioxidant (radical scavenging and superoxide dismutase (SOD)) and antimicrobial activities. Most compounds exhibited antioxidant activities in which compound <strong>15</strong> displayed the most potent radical scavenging and SOD activities. Quantitative structure-activity relationship (QSAR) has been studied using multiple linear regression. The constructed QSAR models displayed high correlation coefficient (Q<sup>2</sup>/<sub><em>Loo-CV</em></sub>= 0.9708 and 0.8753 for RSA and SOD activities, respectively), but low root mean square error (<em>RMSE<sub>LOO-CV </sub>= </em>0.5105 and 1.3571 for RSA and SOD activities, respectively). The structure-activity relationship showed that an ethylene group connected to pyridine ring provided significant antioxidant activities. The QSAR models give insight into the rational designed of eighty new sulfonamides with various electron donating and withdrawing groups. The top five new designed sulfonamides with nitro group are potential antioxidants to be further developed for medicinal applications.</p> Apilak Worachartcheewan Somchai Pisutjaroenpong Ratchanok Pingaew Supaluk Prachayasittikul Suphakit Siriwong Somsak Ruchirawat Virapong Prachayasittikul Copyright (c) 2022 Apilak Worachartcheewan, Somchai Pisutjaroenpong, Ratchanok Pingaew, Supaluk Prachayasittikul, Suphakit Siriwong, Somsak Ruchirawat, Virapong Prachayasittikul http://creativecommons.org/licenses/by/4.0 2022-02-01 2022-02-01 21 360 379 10.17179/excli2021-4590 How to keep up with the analysis of classic and emerging neurotoxins: Age-resolved fitness tests in the animal model Caenorhabditis elegans https://www.excli.de/index.php/excli/article/view/4626 <p>The global chemical inventory includes neurotoxins that are mostly interrogated concerning the biological response in developing organisms. Effects of pollutants on adults receive less attention, although vulnerabilities can be expected throughout the entire life span in young, middle-aged and old individuals. We use the animal model <em>Caenorhabditis elegans</em> to systematically quantify neurological outcomes by application of an age-resolved method. Adult hermaphrodite worms were exposed to pollutants or non-chemical stressors such as temperature in liquid culture on microtiter plates and locomotion fitness was analyzed in a whole-life approach. Cultivation at 15, 20 or 25 °C showed that worms held at 15 °C displayed an enhanced level of fitness concerning swimming movements until middle age (11-days-old) and then a decline. In contrast, <em>C. elegans</em> cultivated at ≥ 20 °C continually reduced their swimming movements with increasing age. Here, we provide a step-by-step protocol to investigate the health span of adult <em>C. elegans</em> that may serve as a platform for automation and data collection. Consistent with this, more neurotoxins can be investigated with respect to vulnerable age-groups as well as contributing non-chemical environmental factors such as temperature.</p> Indra Hering Dang Tri Le Anna von Mikecz Copyright (c) 2022 Indra Hering, Dang Tri Le, Anna von Mikecz http://creativecommons.org/licenses/by/4.0 2022-01-31 2022-01-31 21 344 353 10.17179/excli2021-4626 Ultrasound–guided posterior quadratus lumborum block for postoperative pain control after minimally invasive radical prostatectomy https://www.excli.de/index.php/excli/article/view/4615 <p class="Abstract"><span lang="EN-US">A minimally invasive approach to radical prostatectomy offers improved ambulation and discharge times. Postoperative pain control is one of the key factors that facilitates rapid recovery. With the aim to assure adequate analgesia and minimize the use of opioids, application of truncal nerve blocks has been proposed in a number of endoscopic procedures. The aim of this double-blind, placebo-controlled study was to evaluate the efficacy of bilateral posterior quadratus lumborum block (pQLB) in alleviating pain and reducing postoperative opioid demand in patients following endoscopic extraperitoneal and laparoscopic prostatectomy. We enrolled 50 patients who were diagnosed with prostate cancer and scheduled for prostatectomy. They were randomized to receive preoperative, ultrasound-guided pQLB with the use of either 30 ml of 0.375 % ropivacaine (ropivacaine group) or 30 ml of 0.9 % NaCl (placebo group). Our primary endpoint was opioid consumption in the first 24 hours after surgery. Secondary endpoints were pain intensity at predefined timepoints and the incidence of nausea and vomiting and pruritus. No differences were detected between the ropivacaine and placebo groups in intravenous oxycodone consumption during the first 24 hours after surgery. Similarly, there were no differences in pain intensity at any of the timepoints assessed. The rate of nausea and vomiting was equal in both groups and pruritus was not observed. Application of bilateral pQLB does not reduce opioid consumption after minimally invasive prostatectomy. </span></p> Bartosz Horosz Katarzyna Białowolska Anna Kociuba Jakub Dobruch Małgorzata Malec-Milewska Copyright (c) 2022 Bartosz Horosz, Katarzyna Białowolska, Anna Kociuba, Jakub Dobruch, Małgorzata Malec-Milewska http://creativecommons.org/licenses/by/4.0 2022-01-27 2022-01-27 21 335 343 10.17179/excli2021-4615 Pupil size as an indicator of cognitive activity in mild Alzheimer’s disease https://www.excli.de/index.php/excli/article/view/4568 <p>It is well established that pupil activity indexes cognitive processing. For instance, research has consistently demonstrated that the pupil reacts to working memory span task performance. However, little is known about pupil reaction to cognitive processing in Alzheimer’s Disease (AD). We thus investigated whether span tasks can modulate pupil size in patients with AD. We invited 24 patients with AD and 24 healthy older adults to perform backward and forward spans, as well as to count aloud in a control condition, while their pupil activity was recorded with eye tracking glasses. In patients with AD, analysis demonstrated larger pupil size during backward spans (<em>M</em> = 2.12, <em>SD</em> = .39) than during forward spans (<em>M</em> = 1.98, <em>SD</em> = .36) [<em>t</em>(23) = 3.22, <em>p </em>= .004], larger pupil size during forward spans than during counting (<em>M</em> = 1.67, <em>SD</em> = .33) [<em>t</em>(23) = 4.75, <em>p </em>&lt; .001], as well as larger pupil size during backward spans than during counting [<em>t</em>(23) = 10.60, <em>p </em>&lt; .001]. In control participants, analysis demonstrated larger pupil size during backward spans (<em>M</em> = 3.36, <em>SD</em> = .49) than during forward spans (<em>M</em> = 2.85, <em>SD</em> = .68) [<em>t</em>(23) = 5.82, <em>p </em>&lt; .001], larger pupil size during forward spans than during counting (<em>M</em> = 2.09, <em>SD</em> = .62) [<em>t</em>(23) = 5.42, &lt; .001], as well as larger pupil size during backward spans than during counting [<em>t</em>(23) = 9.70, <em>p </em>&lt; .001]. Results also demonstrated a significant interaction effect between groups and conditions [<em>F</em>(2,92) = 16.63, <em>p </em>&lt; .001]; in other words, patients with AD have shown fewer variations on the pupil size across the conditions compared to the control participants. The larger pupil size during backward spans, compared with forward spans or counting, can be attributed to the high cognitive load of backward spans. The modulation of pupil size, as observed across backward/forward spans and counting, can possibly be attributed to sympathetic/adrenergic and parasympathetic/cholinergic activities. Our study demonstrates the value of pupillometry as a potential biomarker of cognitive processing in AD.</p> Mohamad El Haj Guillaume Chapelet Ahmed A. Moustafa Claire Boutoleau-Bretonnière Copyright (c) 2022 Mohamad El Haj, Guillaume Chapelet, Ahmed A. Moustafa, Claire Boutoleau-Bretonnière http://creativecommons.org/licenses/by/4.0 2022-01-17 2022-01-17 21 307 316 10.17179/excli2021-4568 Inconsistent eating time is associated with obesity https://www.excli.de/index.php/excli/article/view/4324 <p>Obesity is characterized by an accumulation of redundant body fat linked to metabolic dysregulation and low-grade systemic inflammation. Lifestyle choices are imperative determining factors of obesity. The contemporary lifestyle is associated with behaviors that disrupt circadian rhythms, impacting metabolic homeostasis. Our animal and human studies suggest that circadian phenotypes could be related to the risk of metabolic dysregulation and obesity. The purpose of this study is to examine the role of inconsistent eating habits on body weight in adults. Individuals who presented for colon cancer screening were enrolled. Subjects received structured questionnaires to capture 7-day eating and sleeping times in a week prospectively. Bodyweight and height were extracted from medical records, and Body Mass Index (BMI) was calculated. Inconsistent eating times were defined as an average difference of &gt;2 hours between the largest meal on weekdays and weekends. Forty-nine of the 61 (80.3 %) individuals enrolled in the study completed the questionnaires. The mean age and standard deviation (SD) were 60.8 (7.9), and 27 (55.1 %) were male. Subjects with inconsistent eating times had a significantly higher BMI (33.8 ± 3.6 SD, n = 9) than subjects who did not (27.5 ± 6.5 SD, n = 40; p = 0.001). The highest BMI was observed in subjects who ate inconsistently and late (35.8 ± 4.6 SD). In this cross-sectional study, time of eating habits was associated with BMI. Controlled cohort studies are needed to determine the potential link between eating time and the risk of obesity in the long term.</p> Darbaz Adnan Jonathan Trinh Faraz Bishehsari Copyright (c) 2022 Darbaz Adnan, Jonathan Trinh, Faraz Bishehsari http://creativecommons.org/licenses/by/4.0 2022-01-14 2022-01-14 21 300 306 10.17179/excli2021-4324 Enhancing the efficacy of albendazole for liver cancer treatment using mesoporous silica nanoparticles https://www.excli.de/index.php/excli/article/view/4491 <p class="Abstract"><span lang="EN-US">The present study aimed to synthesize albendazole (ABZ)-loaded Mobil Composition of Matter No. 41 (MCM-41 NPs) to increase the efficacy of the drug against liver cancer. ABZ was loaded into MCM-41 NPs, and after <em>in vitro</em> characterization, such as size, size distribution, zeta potential, morphology, chemical composition, thermal profile, drug release, surface and pore volume, and pore size, their biological effects were evaluated using 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) cell migration assays. The results demonstrated that monodispersed and spherical NPs with a size of 220 ± 11.5 and 293 ± 8.7 nm, for MCM-41 NPs and ABZ-loaded MCM-41 NPs, respectively, and drug loading efficiency of 30 % were synthesized. ABZ was loaded physically into MCM-41 NPs, leading to a decrease in surface volume, pore size, and pore volume. Also, MCM-41 NPs could increase the cytotoxicity effects of ABZ by 2.9-fold (IC<sub>50</sub> = 23 and 7.9 µM for ABZ and ABZ-loaded MCM-41 NPs, respectively). In addition, both ABZ and ABZ-loaded MCM-41 NPs could restrain the cell migration by 12 %. Overall, the results of the present study suggest evaluating the potency of MCM-41 NPs, as a potent nanoplatform, for ABZ delivery <em>in vivo</em> environment.</span></p> Mohsen Ghaferi Warda Zahra Azim Akbarzadeh Hasan Ebrahimi Shahmabadi Seyed Ebrahim Alavi Copyright (c) 2022 Copyright will be assigned automatically to Mohsen Ghaferi, Warda Zahra, Azim Akbarzadeh, Hasan Ebrahimi Shahmabadi, Seyed Ebrahim Alavi when this is published http://creativecommons.org/licenses/by/4.0 2022-01-11 2022-01-11 21 236 249 10.17179/excli2021-4491 Preclinical safety profile of a liver-localized mitochondrial uncoupler https://www.excli.de/index.php/excli/article/view/4414 <p class="Abstract"><span lang="EN-US">Mitochondrial uncouplers (mUncouplers) are known to exhibit a variety of toxic effects in animals. Here we report a safety profile of an mUncoupler, OPC-163493, recently synthesized at Otsuka Pharmaceutical Co, Ltd, and its development as a therapeutic agent for treating diabetes. To understand the acute and subchronic toxicity of OPC-163493, single and repeated oral dose studies in rats, dogs, and monkeys were performed. In the rat studies, rigor mortis and increased body temperatures were observed in the high dose group. Focal necrosis, fatty change, and granular eosinophilic cytoplasm of the hepatocytes were also observed in the high dose group. In the dog studies, gastrointestinal manifestations were observed with decreased body weight and decreased food consumption in the high dose group. Necrotizing arteritis was observed in multiple organs as well as meningitis with hemorrhage in the brain. In the monkey studies, vomiting, decreased food consumption, and decreased locomotor activity were observed in the high dose group. Degeneration of the proximal convoluted tubules and the straight tubular epithelium, regeneration of the proximal tubular epithelium, and degeneration of the collecting tubular epithelium were observed. The target organs of OPC-163493 were liver, blood vessels, and kidney in rats, dogs, and monkeys, respectively. In rats, dogs, and monkeys, safety ratios were 100:1, 13:1, and 20:1, respectively, in terms of total exposure (AUC<sub>24h</sub>). These safety ratios showed clear separation between exposure to OPC-163493 in animals at NOAEL and the exposure at the effective dose in ZDF rats. This information should contribute to the drug development of new and effective mUncoupler candidates.</span></p> Yuki Inoue Junichi Kino Nobuya Ishiharada Makoto Sato Suguru Hatanaka Hiroyuki Yokoi Takahiro Shimada Seiji Sato Takashi Okamoto Naohide Kanemoto Copyright (c) 2022 Yuki Inoue, Junichi Kino, Nobuya Ishiharada, Makoto Sato, Suguru Hatanaka, Hiroyuki Yokoi, Takahiro Shimada, Seiji Sato, Takashi Okamoto, Naohide Kanemoto http://creativecommons.org/licenses/by/4.0 2022-01-11 2022-01-11 21 213 235 10.17179/excli2021-4414 Cyclophosphamide and epirubicin induce high apoptosis in microglia cells while epirubicin provokes DNA damage and microglial activation at sub-lethal concentrations https://www.excli.de/index.php/excli/article/view/4160 <p class="Abstract"><span lang="EN-US">Chemotherapy Related Cognitive Impairment (CRCI), also called chemobrain, diminishes cancer patient’s life quality. Breast cancer (BC) patients have been described to be importantly affected, however, the mechanism leading to CRCI has not been fully elucidated. Recent research proposes microglia as the main architect of CRCI, thus dysregulations in these cells could trigger CRCI. The aim of this research was to evaluate the effects of two drugs commonly used against breast cancer, cyclophosphamide (CTX) and epirubicin (EPI), on the microglia cell line SIM-A9, using the BC cell line, 4T1, as a control. Our results show that CTX and EPI decrease microglia-cell viability and increase cell death on a concentration-dependent manner, being 5 and 2 times more cytotoxic to microglia cell line than to breast cancer 4T1cells, respectively. Both chemotherapies induce cell cycle arrest and a significant increase in p53, p16<span style="background: white;"> and γ-H2AX in breast cancer and microglia cells. Furthermore, mitochondrial membrane potential (<span style="color: #222222;">ΔΨ</span></span><span style="color: #222222;">m)</span> diminishes as cell death increases, and both chemotherapies induce reactive oxygen species (ROS) production on SIM-A9 and 4T1. Moreover, caspase activation increases with treatments and its pharmacological blockade inhibits CTX and EPI induced-cell death. Finally, low concentrations of CTX and EPI induce <span style="background: white;">γ-H2AX, and EPI induces cytokine release, NO production and Iba-1 overexpression.</span> These findings indicate that microglia cells are more sensitive to CTX and EPI than BC cells and undergo DNA damage and cell cycle arrest at very low concentrations, moreover EPI induces microglia activation and a pro-inflammatory profile.</span></p> Rafael de la Hoz-Camacho Ana Luisa Rivera-Lazarin Jose Manuel Vazquez-Guillen Diana Caballero-Hernandez Edgar Mendoza-Gamboa Ana Carolina Martinez-Torres Cristina Rodriguez-Padilla Copyright (c) 2022 Rafael de la Hoz-Camacho, Ana Luisa Rivera-Lazarin, Jose Manuel Vazquez-Guillen, Diana Caballero-Hernandez, Edgar Mendoza-Gamboa, Ana Carolina Martinez-Torres, Cristina Rodriguez-Padilla http://creativecommons.org/licenses/by/4.0 2022-01-10 2022-01-10 21 197 212 10.17179/excli2021-4160 Continous, non-invasive monitoring of oxygen consumption in a parallelized microfluidic in vitro system provides novel insight into the response to nutrients and drugs of primary human hepatocytes https://www.excli.de/index.php/excli/article/view/4351 <p class="Abstract"><span lang="EN-US">Oxygen plays a fundamental role in cellular energy metabolism, differentiation and cell biology in general. Consequently, <em>in vitro</em> oxygen sensing can be used to assess cell vitality and detect specific mechanisms of toxicity. In 2D <em>in vitro</em> models currently used, the oxygen supply provided by diffusion is generally too low, especially for cells having a high oxygen demand. In organ-on-chip systems, a more physiologic oxygen supply can be generated by establishing unidirectional perfusion. We established oxygen sensors <em>in</em> an easy-to-use and parallelized organ-on-chip system. We demonstrated the applicability of this system by analyzing the influence of fructose (40 mM, 80 mM), ammonium chloride (100 mM) and Na-diclofenac (50 µM, 150 µM, 450 µM, 1500 µM) on primary human hepatocytes (PHH). Fructose treatment for two hours showed an immediate drop of oxygen consumption (OC) with subsequent increase to nearly initial levels. Treatment with 80 mM glucose, 20 mM lactate or 20 mM glycerol did not result in any changes in OC which demonstrates a specific effect of fructose. Application of ammonium chloride for two hours did not show any immediate effects on OC, but qualitatively changed the cellular response to FCCP treatment. Na-diclofenac treatment for 24 hours led to a decrease of the maximal respiration and reserve capacity. We also demonstrated the stability of our system by repeatedly treating cells with 40 mM fructose, which led to similar cell responses on the same day as well as on subsequent days. In conclusion, our system enables in depth analysis of cellular respiration after substrate treatment in an unidirectional perfused organ-on-chip system.</span></p> <p>in an unidirectional perfused organ-on-chip system.</p> Marius Busche Dominik Rabl Jan Fischer Christian Schmees Torsten Mayr Rolf Gebhardt Martin Stelzle Copyright (c) 2022 Marius Busche, Dominik Rabl, Jan Fischer, Christian Schmees, Torsten Mayr, Rolf Gebhardt, Martin Stelzle http://creativecommons.org/licenses/by/4.0 2022-01-07 2022-01-07 21 144 161 10.17179/excli2021-4351 Hypermethylation of RAD9A intron 2 in childhood cancer patients, leukemia and tumor cell lines suggest a role for oncogenic transformation https://www.excli.de/index.php/excli/article/view/4482 <p>Most childhood cancers occur sporadically and cannot be explained by an inherited mutation or an unhealthy lifestyle. However, risk factors might trigger the oncogenic transformation of cells. Among other regulatory signals, hypermethylation of <em>RAD9A</em> intron 2 is responsible for the increased expression of RAD9A protein, which may play a role in oncogenic transformation. Here, we analyzed the <em>RAD9A</em> intron 2 methylation in primary fibroblasts of 20 patients with primary cancer in childhood and second primary cancer (2N) later in life, 20 matched patients with only one primary cancer in childhood (1N) and 20 matched cancer-free controls (0N), using bisulfite pyrosequencing and deep bisulfite sequencing (DBS). Four 1N patients and one 2N patient displayed elevated mean methylation levels (³10 %) of <em>RAD9A</em>. DBS revealed ³2 % hypermethylated alleles of <em>RAD9A,</em> indicative for constitutive mosaic epimutations. Bone marrow samples of NHL and AML tumor patients (n=74), EBV (Epstein Barr Virus) lymphoblasts (n=6), tumor cell lines (n=5) and FaDu subclones (n=13) were analyzed to substantiate our findings. We find a broad spectrum of tumor entities with an aberrant methylation of <em>RAD9A</em>. We detected a significant difference in mean methylation of <em>RAD9A</em> for NHL versus AML patients (p ≤0.025). Molecular karyotyping of AML samples during therapy with hypermethylated <em>RAD9A</em> showed an evolving duplication of 1.8 kb on Chr16p13.3 including the PKD1 gene. Radiation, colony formation assays, cell proliferation, PCR and molecular karyotyping SNP-array experiments using generated FaDu subclones suggest that hypermethylation of <em>RAD9A</em> intron 2 is associated with genomic imbalances in regions with tumor-relevant genes and survival of the cells. In conclusion, this is the very first study of <em>RAD9A</em> intron 2 methylation in childhood cancer and Leukemia. <em>RAD9A</em> epimutations may have an impact on leukemia and tumorigenesis and can potentially serve as a biomarker.</p> Danuta Galetzka Julia Böck Lukas Wagner Marcus Dittrich Olesja Sinizyn Marco Ludwig Heidi Rossmann Claudia Spix Markus Radsak Peter Scholz-Kreisel Johanna Mirsch Matthias Linke Walburgis Brenner Manuela Marron Alicia Poplawski Thomas Haaf Heinz Schmidberger Dirk Prawitt Copyright (c) 2022 Danuta Galetzka, Julia Böck, Lukas Wagner, Marcus Dittrich, Olesja Sinizyn , Marco Ludwig, Heidi Rossmann, Claudia Spix, Markus Radsak, Peter Scholz-Kreisel, Johanna Mirsch, Matthias Linke, Walburgis Brenner, Manuela Marron, Alicia Poplawski, Thomas Haaf, Heinz Schmidberger, Dirk Prawitt http://creativecommons.org/licenses/by/4.0 2022-01-07 2022-01-07 21 117 143 10.17179/excli2021-4482 Prevalence and predictors of adequate treatment of overt hypothyroidism – a population-based study https://www.excli.de/index.php/excli/article/view/4291 <p>The aim of this study is to evaluate the adequacy of treatment, and to identify factors influencing treatment of hypothyroidism. Patients newly diagnosed with overt hypothyroidism (<em>n</em>=345) were identified via a register linked to a laboratory database. In selected periods with staff available, 165 patients were invited, and 113 (68.5 %) accepted participating in a comprehensive program including blood tests and completion of questionnaires. We performed a longitudinal follow-up on thyroid function tests 10 years after the diagnosis. Time to reach a serum TSH level of 0.2-10 mU/L (termed as clinically acceptable) and biochemical normalization (TSH: 0.2-5.0 mU/L), respectively, were analyzed using Kaplan Meier survival analysis. Predictors for longer duration to reach the normal TSH range were identified using cox proportional hazards regression. Only 67.7 % of the patients were in the euthyroid range on the long term after diagnosis of overt hypothyroidism (2 years: 59.4 %; 10 years: 67.7 %). Median time to the first normal TSH was 8.9 months (95 % CI: 7.6-10.2 months). The factors associated with longer duration until normalization of TSH after multivariate analysis were age (HR 0.79 per 10 years; 95 % CI: 0.66-0.94; <em>P </em>= &lt;0.01), smoking (HR 0.47; 95 % CI: 0.26-0.83; <em>P </em>= &lt;0.01), serum TSH at diagnosis (HR 0.96 per 10 mU/L; 95 % CI: 0.93-0.99; <em>P </em>= 0.02) and BMI (HR 0.96 per kg/m<sup>2</sup>; 95 % CI: 0.91-0.99; <em>P </em>= 0.03). A considerable number of hypothyroid patients remained inadequately treated. When treating hypothyroid patients, special attention should be addressed to those patients who never or lately obtain euthyroid status.</p> Julie Lindgård Nielsen Jesper Karmisholt Inge Bülow Pedersen Allan Carlé Copyright (c) 2022 Julie Lindgård Nielsen, Jesper Karmisholt, Inge Bülow Pedersen, Allan Carlé http://creativecommons.org/licenses/by/4.0 2022-01-06 2022-01-06 21 104 116 10.17179/excli2021-4291 The COVID-19 vaccination acceptance/hesitancy rate and its determinants among healthcare workers of 91 Countries https://www.excli.de/index.php/excli/article/view/4439 <p>The aim of this study was to investigate the COVID-19 vaccination acceptance rate and its determinants among healthcare workers in a multicenter study. This was a cross-sectional multi-center survey conducted from February 5 to April 29, 2021<em>.</em> The questionnaire consisted of 26 items in 6 subscales. The English version of the questionnaire was translated into seven languages and distributed through Google Forms using snowball sampling; a colleague in each country was responsible for the forward and backward translation, and also the distribution of the questionnaire. A forward stepwise logistic regression was utilized to explore the variables and questionnaire factors tied to the intention to COVID-19 vaccination. 4630 participants from 91 countries completed the questionnaire. According to the United Nations Development Program 2020, 43.6 % of participants were from low Human Development Index (HDI) regions, 48.3 % high and very high, and 8.1 % from medium. The overall vaccination hesitancy rate was 37 %. Three out of six factors of the questionnaire were significantly related to intention to the vaccination. While ‘Perceived benefits of the COVID-19 vaccination’ (OR: 3.82, p-value&lt;0.001) and ‘Prosocial norms’ (OR: 5.18, p-value&lt;0.001) were associated with vaccination acceptance, ‘The vaccine safety/cost concerns’ with OR: 3.52, p-value&lt;0.001 was tied to vaccination hesitancy. Medical doctors and pharmacists were more willing to take the vaccine in comparison to others. Importantly, HDI with OR: 12.28, 95 % CI: 6.10-24.72 was a strong positive determinant of COVID-19 vaccination acceptance. This study highlighted the vaccination hesitancy rate of 37 % in our sample among HCWs. Increasing awareness regarding vaccination benefits, confronting the misinformation, and strengthening the prosocial norms would be the primary domains for maximizing the vaccination coverage. The study also showed that the HDI is strongly associated with the vaccination acceptance/hesitancy, in a way that those living in low HDI contexts are more hesitant to receive the vaccine.</p> Mehrdad Askarian Aleksandr Semenov Ferran Llopis Francesca Rubulotta Gorana Dragovac Natalia Pshenichnaya Ojan Assadian Yvon Ruch Zahra Shayan Cristobal Padilla Fortunatti Daniel Lucey Abdullah Almohaizeie Abu Hena Mostafa Kamal Adenike Ogunshe Aidos Konkayev Asim Beg Enzo Primerano Fatma Amer Hema Prakash Kumari Pilli Ivan Hung Folusakin Ayoade Jean Yves Lefrant Joanna Zajkowska Jordi Rello Momin Kazi Mohammad Hossein Taghrir Stijn Blot Stephen Leib Parisa Hosseinpour Hamidreza Hosseinpour Amirhossein Erfani Roham Borazjani Hossein Akbarialiabad Masoud Najafi Ardalan Askarian Hakan Erdem Copyright (c) 2022 Mehrdad Askarian, Aleksandr Semenov, Ferran Llopis, Francesca Rubulotta, Gorana Dragovac, Natalia Pshenichnaya, Ojan Assadian, Yvon Ruch, Zahra Shayan, Cristobal Padilla Fortunatti, Daniel Lucey, Abdullah Almohaizeie, Abu Hena Mostafa Kamal, Adenike Ogunshe, Aidos Konkayev, Asim Beg, Enzo Primerano, Fatma Amer, Hema Prakash Kumari Pilli, Ivan Hung, Folusakin Ayoade, Jean Yves Lefrant, Joanna Zajkowska, Jordi Rello, Momin Kazi, Mohammad Hossein Taghrir, Stijn Blot, Stephen Leib, Parisa Hosseinpour, Hamidreza Hosseinpour, Amirhossein Erfani, Roham Borazjani, Hossein Akbarialiabad, Masoud Najafi, Ardalan Askarian, Hakan Erdem http://creativecommons.org/licenses/by/4.0 2022-01-06 2022-01-06 21 93 103 10.17179/excli2021-4439 Feeding to satiation induces mild oxidative/carbonyl stress in the brain of young mice https://www.excli.de/index.php/excli/article/view/4347 <p>Intermittent fasting as a dietary intervention can prevent overweight and obesity in adult organisms. Nevertheless, information regarding consequences of intermittent fasting for redox status and reactive metabolite-mediated processes that are crucial for the normal functioning of organisms is limited. Since the information on effects of intermittent fasting on parameters of oxidative/carbonyl stress in the brains of young mice was absent, the present study addressed these questions using an every-other-day fasting (EODF) protocol. The levels of carbonyl proteins were ~28 %, 22 % and 18 % lower in the cerebral cortex of EODF males and females and middle parts of the brain of EODF males, respectively, as compared to their <em>ad libitum</em> fed counterparts. Lipid peroxides and α-dicarbonyl compounds were lower only in the cortex and medulla part of EODF male brain. The EODF regimen resulted in higher total non-specific antioxidant capacity in different parts of male brain and a tendency to be higher this parameter in females. At the same time, EODF regimen had no effect on the activities of the defensive antioxidant enzymes, namely superoxide dismutase, catalase, glutathione-S-transferase, glutathione peroxidase, glyoxylase 1 and glucose-6-phosphate dehydrogenase in the cortex of both sexes, but even decreased activities of these enzymes in medulla and middle part of the brain. In general, the results suggest that in the brain of young mice <em>ad libitum</em> feeding induces mild oxidative/carbonyl stress which may be partially alleviated by the EODF regimen. The effect of EODF regimen is more pronounced in the medulla part than in the cortex.</p> Oksana V. Kuzniak Oksana M. Sorochynska Maria M. Bayliak Andrii Ya. Klonovskyi Yulia V. Vasylyk Halyna M. Semchyshyn Kenneth B. Storey Olga Garaschuk Volodymyr I. Lushchak Copyright (c) 2022 Oksana V. Kuzniak, Oksana M. Sorochynska, Maria M. Bayliak, Andrii Ya. Klonovskyi, Yulia V. Vasylyk, Halyna M. Semchyshyn, Kenneth B. Storey, Olga Garaschuk, Volodymyr I. Lushchak http://creativecommons.org/licenses/by/4.0 2022-01-05 2022-01-05 21 77 92 10.17179/excli2021-4347 Risk factors, time to onset and recurrence of delirium in a mixed medical-surgical ICU population https://www.excli.de/index.php/excli/article/view/4381 <p>A retrospective secondary analysis of 4,200 patients was collected from two academic medical centers. Delirium was assessed using the Confusion Assessment Method for the Intensive Care Unit (CAM-ICU) in all patients. Univariate and multivariate Cox models, logistic regression analysis, and Chi-square Automatic Interaction Detector (CHAID) decision tree modeling were used to explore delirium risk factors. Increased delirium risk was associated with exposed only to artificial light (AL) hazard ratio (HR) 1.84 (95 % CI: 1.66-2.044, <em>P</em>&lt;0.001), physical restraint application 1.11 (95 % CI: 1.001-1.226, <em>P</em>=0.049), and high nursing care requirements (&gt;8 hours per 8-hour shift) 1.18 (95 % CI: 1.048-1.338, <em>P</em>=0.007). Delirium incidence was inversely associated with greater family engagement 0.092 (95 % CI: 0.014-0.596, <em>P</em>=0.012), low staff burnout and anticipated turnover scores 0.093 (95 % CI: 0.014-0.600, <em>P</em>=0.013), non-ICU length-of-stay (LOS)&lt;15 days 0.725 (95 % CI: 0.655-0.804, <em>P</em>&lt;0.001), and ICU LOS ≤15 days 0.509 (95 % CI: 0.456-0.567, <em>P</em>&lt;0.001). CHAID modeling indicated that AL exposure and age &lt;65 years were associated with a high risk of delirium incidence, whereas SOFA score ≤11, APACHE IV score &gt;15 and natural light (NL) exposure were associated with moderate risk, and female sex was associated with low risk. More rapid time to delirium onset correlated with baseline sleep disturbance (<em>P</em>=0.049), high nursing care requirements (<em>P</em>=0.019), and prolonged ICU and non-ICU hospital LOS (<em>P</em>&lt;0.001). Delirium recurrence correlated with age &gt;65 years (HR 2.198; 95 % CI: 1.101-4.388, <em>P</em>=0.026) and high nursing care requirements (HR 1.978, 95 % CI: 1.096-3.569), with CHAID modeling identifying AL exposure (<em>P</em>&lt;0.001) and age &gt;65 years (<em>P</em>=0.032) as predictive variables. Development of ICU delirium correlated with application of physical restraints, high nursing care requirements, prolonged ICU and non-ICU LOS, exposure exclusively to AL (rather than natural), less family engagement, and greater staff burnout and anticipated turnover scores. ICU delirium occurred more rapidly in patients with baseline sleep disturbance, and recurrence correlated with the presence of delirium on ICU admission, exclusive AL exposure, and high nursing care requirements.</p> Farshid Rahimibashar Andrew C. Miller Mahmood Salesi Motahareh Bagheri Amir Vahedian-Azimi Sara Ashtari Keivan Gohari Moghadam Amirhossein Sahebkar Copyright (c) 2022 Farshid Rahimibashar, Andrew C. Miller, Mahmood Salesi, Motahareh Bagheri, Amir Vahedian-Azimi, Sara Ashtari, Keivan Gohari Moghadam, Amirhossein Sahebkar http://creativecommons.org/licenses/by/4.0 2022-01-04 2022-01-04 21 30 46 10.17179/excli2021-4381 Sex-specific differences in trimethylamine N-oxide (TMAO) concentrations before and after cardiac rehabilitation in acute myocardial infarction patients https://www.excli.de/index.php/excli/article/view/4366 <p class="Abstract"><span lang="EN-US">Trimethylamine N-oxide (TMAO) is a biomarker of cardiovascular risk and may enhance the progression of atherosclerosis. <a name="_Hlk62595083"></a>The aim of the study was to determine whether there are sex-specific differences in TMAO concentrations before and after cardiac rehabilitation in acute myocardial infarction (AMI) patients. A total of 56 participants [45/56 (80.4 %) males, 11/56 (19.6 %) females] were drawn from AMI inpatients hospitalized at the Division of Cardiology, Medical University of Graz, Austria. For the assessment of TMAO, serum samples were collected within the first day after hospital admission due to AMI and at the start and end of cardiac rehabilitation. Shortly after hospital admission due to AMI, females had significantly higher TMAO blood concentrations than males. These initially high TMAO levels remained almost unchanged in the female AMI patients until the start of cardiac rehabilitation and only reached the lower TMAO concentrations <a name="_Hlk73174718"></a>observed in the male patients after rehabilitation [female patients: TMAO (acute myocardial infarction) = 5.93 μmol/L (SE = 1.835); TMAO (start of rehabilitation) = 5.68 μmol/L (SE = 1.217); TMAO (end of rehabilitation) = 3.89 μmol/L (SE = 0.554); male patients: TMAO (acute myocardial infarction) = 3.02 μmol/L (SE = 0.255), TMAO (start of rehabilitation) = 3.91 μmol/L (SE = 0.346), TMAO (end of rehabilitation) = 4.04 μmol/L (SE = 0.363)]. After AMI, women might be at higher cardiovascular risk due to persistently higher levels of TMAO. High TMAO levels in women might decrease after cardiac rehabilitation due to cardiac rehabilitation-associated lifestyle modifications. These lifestyle modifications after AMI might also prevent increases in TMAO concentrations in men.</span></p> Andreas Baranyi Andreas Meinitzer Dirk von Lewinski Hans-Bernd Rothenhäusler Omid Amouzadeh-Ghadikolai Hanns Harpf Leonhard Harpf Heimo Traninger Ronald Hödl Birgit M. Harb Barbara Obermayer-Pietsch Melanie Schweinzer Celine K. Braun Dietmar Enko Copyright (c) 2022 Andreas Baranyi, Andreas Meinitzer, Dirk von Lewinski, Hans-Bernd Rothenhäusler, Omid Amouzadeh-Ghadikolai, Hanns Harpf, Leonhard Harpf, Heimo Traninger, Ronald Hödl, Birgit M. Harb, Barbara Obermayer-Pietsch, Melanie Schweinzer, Celine K. Braun, Dietmar Enko http://creativecommons.org/licenses/by/4.0 2022-01-03 2022-01-03 21 1 10 10.17179/excli2021-4366 Using R in regulatory toxicology https://www.excli.de/index.php/excli/article/view/5097 <p>Statistical analyses are an essential part of regulatory toxicological evaluations. While projects would be ideally monitored by both toxicologists and statisticians, this is often not possible in practice. Hence, toxicologists should be trained in some common statistical approaches but also need a tool for statistical evaluations. Due to transparency needed in regulatory processes and standard tests that can be evaluated with template approaches, the freely available open-source statistical software <strong>R</strong> may be suitable. <strong>R</strong> is a well-established software in the statistical community. The principal input method is via software code, which is both benefit and weakness of the tool. It is increasingly used by regulating authorities globally and can be easily extended by software packages, e.g., for new statistical functions and features. This manuscript outlines how <strong>R</strong> can be used in regulatory toxicology, allowing toxicologists to perform all regulatory required data evaluations in a single software solution. Practical applications are shown in case studies on simulated and experimental data. The examples cover a) Dunnett testing of treatment groups against a common control and in relation to a biological relevance threshold, assessing the test’s assumptions and plotting the results; b) dose-response analysis and benchmark dose derivation for chronic kidney inflammation as a function of Pyridine; and c) graphical/exploratory data analysis of previously published developmental neurotoxicity data for Chlorpyrifos.</p> Felix M. Kluxen Signe M. Jensen Copyright (c) 2022 Felix M. Kluxen, Signe M. Jensen http://creativecommons.org/licenses/by/4.0 2022-08-22 2022-08-22 21 1130 1150 10.17179/excli2022-5097 Clinical presentation vs endoscopy for an early diagnosis of eosinophilic esophagitis https://www.excli.de/index.php/excli/article/view/4838 <p class="Abstract"><span lang="EN-US">Eosinophilic esophagitis (EoE) is a type-2 mediated, chronic inflammatory disease showing an increase of both incidence and prevalence. Early diagnosis is mandatory, to prevent fibrostenotic complication of the disease. Due to the low sensitivity of the classic endoscopic features of the disease, a strong clinical suspicion should drive the decision to collect mucosal biopsies of the esophagus. We describe the case of an atopic patient suffering from dysphagia with normal esophageal mucosa and frank histological hallmarks of the disease. </span></p> Michele Di Stefano Giacomo Grandi Vera Bonaso Elisabetta Pagani Gino Roberto Corazza Antonio Di Sabatino Copyright (c) 2022 Michele Di Stefano, Giacomo Grandi, Vera Bonaso, Elisabetta Pagani, Gino Roberto Corazza, Antonio Di Sabatino http://creativecommons.org/licenses/by/4.0 2022-03-24 2022-03-24 21 651 655 10.17179/excli2022-4838 Use of intranasal esketamine in a girl with treatment-resistant depression and autism spectrum disorders https://www.excli.de/index.php/excli/article/view/4694 <p class="Abstract"><span lang="EN-US">Major depression is a common comorbidity in autism spectrum disorder (ASD), often difficult to identify and to treat. Autistic subjects are more at risk for suicidal thoughts and behaviors compared to typically developing peers. Unfortunately, ASD individuals are more frequently treatment-resistant and often show side-effects which reduce efficacy. Intranasal esketamine has been recently approved as an add-on medication for treatment-resistant depression (TRD), but it has never been used in ASD with comorbid major depression. Of note, a pilot study of intranasal ketamine has shown no effect on social withdrawal in ASD without depression. The present case report describes the first girl with ASD and comorbid TRD treated with intranasal esketamine.</span></p> Miriam Olivola Vincenzo Arienti Nicola Bassetti Serena Civardi Natascia Brondino Copyright (c) 2022 Miriam Olivola, Vincenzo Arienti, Nicola Bassetti, Serena Civardi, Natascia Brondino http://creativecommons.org/licenses/by/4.0 2022-02-28 2022-02-28 21 540 543 10.17179/excli2022-4694 Kinetics of humoral immune response in patients with asymptomatic or mild COVID-19 https://www.excli.de/index.php/excli/article/view/5337 Nathanielly de Lima Silva Danilo Nobre Joyceane Alves de Oliveira Márcia Santos Rezende Joyce Thayane da Conceição dos Santos Adriano Antunes de Souza Araújo Lucindo José Quintans-Júnior Rafael Ciro Marques Cavalcante Luiz Carlos de Souza Ferreira Paulo Ricardo Martins-Filho Dulce Marta Schimieguel Copyright (c) 2022 Nathanielly de Lima Silva, Danilo Nobre, Joyceane Alves de Oliveira, Márcia Santos Rezende, Joyce Thayane da Conceição dos Santos, Adriano Antunes de Souza Araújo, Lucindo José Quintans-Júnior, Rafael Ciro Marques Cavalcante, Luiz Carlos de Souza Ferreira, Paulo Ricardo Martins-Filho, Dulce Marta Schimieguel http://creativecommons.org/licenses/by/4.0 2022-08-26 2022-08-26 21 1167 1170 10.17179/excli2022-5337 Comment to the Letter to the Editor entitled “Lithium in drinking water and suicide risk” by Tomoyuki Kawada https://www.excli.de/index.php/excli/article/view/5227 Antonina Luca Maria Luca Copyright (c) 2022 Antonina Luca, Maria Luca http://creativecommons.org/licenses/by/4.0 2022-08-03 2022-08-03 21 1066 1067 10.17179/excli2022-5227 Recent insights into the biological functions of baicalin https://www.excli.de/index.php/excli/article/view/5184 Priscilla Nadalin Jae Kwang Kim Tae Won Kim Sang Un Park Copyright (c) 2022 Priscilla Nadalin, Jae Kwang Kim, Tae Won Kim, Sang Un Park http://creativecommons.org/licenses/by/4.0 2022-08-01 2022-08-01 21 1019 1027 10.17179/excli2022-5184 Delayed skin reactions after the second dose of mRNA vaccines against SARS-CoV-2 https://www.excli.de/index.php/excli/article/view/5216 Paulo Ricardo Martins-Filho Martha Débora Lira Tenório Copyright (c) 2022 Paulo Ricardo Martins-Filho, Martha Débora Lira Tenório http://creativecommons.org/licenses/by/4.0 2022-07-27 2022-07-27 21 1015 1018 10.17179/excli2022-5216 Interventions on soy isoflavone molecules to improve their therapeutic potential for prostate cancer treatment https://www.excli.de/index.php/excli/article/view/5130 Vladimir Ajdžanović Branka Šošić-Jurjević Vesna Vodnik Branko Filipović Copyright (c) 2022 Vladimir Ajdžanović, Branka Šošić-Jurjević, Vesna Vodnik, Branko Filipović http://creativecommons.org/licenses/by/4.0 2022-07-01 2022-07-01 21 941 947 10.17179/excli2022-5130 Need to focus on inhibitory activity of benzimidazole analogues against indolamine 2,3-dioxygenase-1 (IDO-1) https://www.excli.de/index.php/excli/article/view/4988 Akshansh Sharma Rajiv Tonk Ravi Shekhar Sushil Dohare Deepak Kumar Copyright (c) 2022 Akshansh Sharma, Rajiv Tonk, Ravi Shekhar, Sushil Dohare, Deepak Kumar http://creativecommons.org/licenses/by/4.0 2022-06-28 2022-06-28 21 904 905 10.17179/excli2022-4988 Current update on the protective effect of epicatechin in neurodegenerative diseases https://www.excli.de/index.php/excli/article/view/5034 Riya Thapa Gaurav Gupta Piyush Dave Santosh Kumar Singh Abhay Raizaday Waleed Hassan Almalki Govind Vyas Sachin Kumar Singh Kamal Dua Yogendra Singh Copyright (c) 2022 Riya Thapa, Gaurav Gupta, Piyush Dave, Santosh Kumar Singh, Abhay Raizaday, Waleed Hassan Almalki, Govind Vyas, Sachin Kumar Singh, Kamal Dua, Yogendra Singh http://creativecommons.org/licenses/by/4.0 2022-06-24 2022-06-24 21 897 903 10.17179/excli2022-5034 Introducing a new index for selecting genetic polymorphisms for association studies https://www.excli.de/index.php/excli/article/view/5004 Nafiseh Omidpanah Mostafa Saadat Copyright (c) 2022 Nafiseh Omidpanah, Mostafa Saadat http://creativecommons.org/licenses/by/4.0 2022-06-10 2022-06-10 21 814 817 10.17179/excli2022-5004 A revolution in health: Opportunities and challenges of the Metaverse https://www.excli.de/index.php/excli/article/view/5017 Milad Ahmadi Marzaleh Mahmoudreza Peyravi Fatemeh Shaygani Copyright (c) 2022 Milad Ahmadi Marzaleh, Mahmoudreza Peyravi, Fatemeh Shaygani http://creativecommons.org/licenses/by/4.0 2022-05-31 2022-05-31 21 791 792 10.17179/excli2022-5017 What are risk factors for COVID-19 vaccine breakthrough infections in patients with previous history of bariatric surgery? https://www.excli.de/index.php/excli/article/view/4873 Gerardo Sarno Pietro Calabrese Luigi Schiavo Francesco Izzo Vincenzo Pilone Copyright (c) 2022 Gerardo Sarno, Pietro Calabrese, Luigi Schiavo, Francesco Izzo, Vincenzo Pilone http://creativecommons.org/licenses/by/4.0 2022-05-12 2022-05-12 21 786 790 10.17179/excli2022-4873 Current update on anticancer effects of icariin https://www.excli.de/index.php/excli/article/view/4848 Mukta Gupta Yachana Mishra Vijay Mishra Murtaza M. Tambuwala Copyright (c) 2022 Mukta Gupta, Yachana Mishra, Vijay Mishra, Murtaza M. Tambuwala http://creativecommons.org/licenses/by/4.0 2022-04-06 2022-04-06 21 680 686 10.17179/excli2022-4848 HDAC11: A novel inflammatory biomarker in Huntington’s disease https://www.excli.de/index.php/excli/article/view/4741 Vishal Kumar Simranjit Kaur Lakshay Kapil Charan Singh Arti Singh Copyright (c) 2022 Vishal Kumar, Simranjit Kaur, Lakshay Kapil, Charan Singh, Arti Singh http://creativecommons.org/licenses/by/4.0 2022-03-24 2022-03-24 21 647 650 10.17179/excli2022-4741 Poorer and more densely populated regions have lower vaccination capability against COVID-19 https://www.excli.de/index.php/excli/article/view/4798 Paulo Ricardo Martins-Filho Ricardo Ruan Rocha Santana Victor Santana Santos Lorena G. Barberia Copyright (c) 2022 Paulo Ricardo Martins-Filho, Ricardo Ruan Rocha Santana, Victor Santana Santos, Lorena G. Barberia http://creativecommons.org/licenses/by/4.0 2022-03-21 2022-03-21 21 621 622 10.17179/excli2022-4798 Current update on the protective effect of naringin in inflammatory lung diseases https://www.excli.de/index.php/excli/article/view/4752 Gaurav Gupta Waleed Hassan Almalki Imran Kazmi Neeraj Kumar Fuloria Shivkanya Fuloria Vetriselvan Subramaniyan Mahendran Sekar Sachin Kumar Singh Dinesh Kumar Chellappan Kamal Dua Copyright (c) 2022 Gaurav Gupta, Waleed Hassan Almalki, Imran Kazmi, Neeraj Kumar Fuloria, Shivkanya Fuloria, Vetriselvan Subramaniyan, Mahendran Sekar, Sachin Kumar Singh, Dinesh Kumar Chellappan, Kamal Dua http://creativecommons.org/licenses/by/4.0 2022-03-07 2022-03-07 21 573 579 10.17179/excli2022-4752 Lithium in drinking water and suicide risk https://www.excli.de/index.php/excli/article/view/4689 Tomoyuki Kawada Copyright (c) 2022 Tomoyuki Kawada http://creativecommons.org/licenses/by/4.0 2022-03-03 2022-03-03 21 571 572 10.17179/excli2022-4689 Toll-like receptors, innate immune system, and lung diseases https://www.excli.de/index.php/excli/article/view/4688 Vyoma K. Patel Keshav R. Paudel Shakti D. Shukla Gang Liu Brian G. Oliver Philip M. Hansbro Kamal Dua Copyright (c) 2022 Vyoma K. Patel, Keshav R. Paudel, Shakti D. Shukla, Gang Liu, Brian G. Oliver, Philip M. Hansbro, Kamal Dua http://creativecommons.org/licenses/by/4.0 2022-02-25 2022-02-25 21 519 523 10.17179/excli2022-4688 Recent insights into the biological and pharmacological activity of lycopene https://www.excli.de/index.php/excli/article/view/4714 Jae Kwang Kim Sang Un Park Copyright (c) 2022 Jae Kwang Kim, Sang Un Park http://creativecommons.org/licenses/by/4.0 2022-02-16 2022-02-16 21 415 425 10.17179/excli2022-4714 Metabolic syndrome in childhood cancer survivors https://www.excli.de/index.php/excli/article/view/3916 Amena Firoz Muhammad Haris Copyright (c) 2022 Amena Firoz, Muhammad Haris http://creativecommons.org/licenses/by/4.0 2022-02-10 2022-02-10 21 380 386 10.17179/excli2021-3916 Preventive role of Sapindus species in different neurological and metabolic disorders https://www.excli.de/index.php/excli/article/view/4625 Sarita Rawat Gaurav Gupta Anurag Mishra Sachchidanand Pathak Lakshmi Thangavelu Sachin Kumar Singh Niraj Kumar Jha Deepak Kumar Poonam Negi Avvaru Praveen Kumar Dinesh Kumar Chellappan Kamal Dua Copyright (c) 2022 Sarita Rawat, Gaurav Gupta, Anurag Mishra, Sachchidanand Pathak, Lakshmi Thangavelu, Sachin Kumar Singh, Niraj Kumar Jha, Deepak Kumar, Poonam Negi, Avvaru Praveen Kumar, Dinesh Kumar Chellappan, Kamal Dua http://creativecommons.org/licenses/by/4.0 2022-01-31 2022-01-31 21 354 359 10.17179/excli2021-4625 Seroprevalence of SARS-CoV-2 antibodies in radio and television workers https://www.excli.de/index.php/excli/article/view/4545 Paulo Ricardo Martins-Filho Danilo Nobre da Silva Danillo Menezes dos Santos Márcia Santos Rezende Jessica Paloma Rosa Silva Josafá Bonifácio da Silva Neto Dulce Marta Schimieguel Lucindo José Quintans-Júnior Jullyana de Souza Siqueira Quintans Paula Santos Nunes Adriano Antunes de Souza Araújo Copyright (c) 2022 Paulo Ricardo Martins-Filho, Danilo Nobre da Silva, Danillo Menezes dos Santos, Márcia Santos Rezende, Jessica Paloma Rosa Silva, Josafá Bonifácio da Silva Neto, Dulce Marta Schimieguel, Lucindo José Quintans-Júnior, Jullyana de Souza Siqueira Quintans, Paula Santos Nunes, Adriano Antunes de Souza Araújo http://creativecommons.org/licenses/by/4.0 2022-01-12 2022-01-12 21 269 272 10.17179/excli2021-4545