bluuebunny/crossref_metadata_2025 huggingface dataset:
Working Hard and Pushing Through: A Thematic Analysis of Humanitarian Migrants’ Experiences in the Australian Workforce
Authors: Cain Patricia , Reid Alison
Published: November 2021

Employment can play an important role for humanitarian migrants in their successful integration into a new country. For humanitarian migrants to Australia, there are no skill or language restrictions imposed on resettlement. Despite the benefits, humanitarian migrants often find themselves in low-status jobs and precarious working conditions. The present study examines perceptions of job quality and exposure to workforce psychosocial risk factors such as job strain, bullying, and discrimination. We conducted semi-structured in-depth interviews with 30 humanitarian migrants from South Sudan, Afghanistan, and Iraq. Thematic analysis of transcripts identified three overarching themes: Uncertainty and Insecurity, Working Hard and Pushing Through, and Positive Attitudes and Actions. Overall, our findings show that despite high levels of education and long-term residency in Australia, many of the participants struggled to find a safe and secure place in the workforce. While some spoke about their work in positive terms, their comments should not be taken as confirmation of a positive work environment. Humanitarian migrants face an uphill battle against oppressive working conditions and underemployment.
Relationship of Binge Drinking with Impairments Affecting Memory and Executive Function among University Students: A Cross-Sectional Study in Northern Spain
Authors: Herrero-Montes Manuel , Alonso-Blanco Cristina , Paz-Zulueta María , Pellico-López Amada , Ruiz-Azcona Laura , Sarabia-Cobo Carmen , Boixadera-Planas Ester , Parás-Bravo Paula
Published: November 2021

Binge drinking (BD) is a common practice among college students. Alcohol consumption may affect brain structures that continue to develop in early adulthood. The aim of this study was to analyze the relationship of BD with impairments affecting memory and executive function among university students. A cross-sectional study was conducted among students (aged 18–30 years) enrolled for the academic year 2018–2019 at the Faculty of Nursing of the University of Cantabria (Spain). Data collection included sociodemographic and academic information, together with alcohol and drug use information, collected by means of a semi-structured questionnaire. A battery of validated tests was used to gather neuropsychological variables. A total of 142 participants were included, of which 88.03% were women. Up to 38.03% were classified as BD. No differences were found in memory tests. Regarding executive function, better performance in the Stroop Color and Word Test was observed in non-BD but the results were not statistically significant. In conclusion, no relationship was found between memory and executive function and BD, although better performance in executive function, specifically inhibitory control, was observed in non-BD.
Relationship between Psychological Status and Health Behaviors during the Coronavirus Disease Pandemic in Japanese Community-Dwelling Older Adults
Authors: Hara Shuichi , Miura Hiroko , Hita Tsuyoshi , Sasaki Sahara , Ito Hidetoshi , Kozaki Yumi , Kawasaki Yoshiko
Published: November 2021

The coronavirus disease (COVID-19) continues to be a widespread pandemic. We investigated the relationship between anxiety/stress and health behaviors during the COVID-19 pandemic in homebound Japanese older adults during January and February 2021. We surveyed 1507 community-dwelling, older Japanese adults using a self-administered questionnaire on primary attributes, including family structure, evaluation of psychological anxiety/stress, and health behaviors. Participants were divided into four anxiety/stress groups based on the frequency of experiencing anxiety/stress, and their association with health behaviors was analyzed using bivariate and multivariate analyses. Responses were received from 469 (31.1%) respondents. In the bivariate analysis, age and family structure were significantly associated with anxiety/stress (p < 0.01). The health behaviors significantly associated with anxiety/stress were walking, balanced eating habits, limited snacking, regular lifestyle, and dental visits. Logistic regression analysis was performed using the variables in the bivariate analysis that showed a significant association with anxiety/stress status as independent variables. Finally, age and dietary habits were significantly associated with anxiety/stress status. No significant associations were found between any other variables. Among older adults living in the rural areas of Japan during the COVID-19 pandemic, anxiety/stress status was significantly associated with age and dietary habits but not with other health behaviors.
Improving the Analysis of E-Cigarette Emissions: Detecting Human “Dry Puff” Conditions in a Laboratory as Validated by a Panel of Experienced Vapers
Authors: Visser Wouter F. , Krüsemann Erna J. Z. , Klerx Walther N. M. , Boer Karin , Weibolt Naomi , Talhout Reinskje
Published: November 2021

Introduction: E-cigarette product regulation requires accurate analyses of emissions. User behavior, including device power setting selection, should be mimicked closely when generating e-cigarette emissions in a laboratory. Excessively high power settings result in an adverse burnt off-taste, called “dry puff flavor”. This should be avoided because it results in an overestimation of toxicant levels (especially certain carbonyls). This study presents a human volunteer-validated approach to detect excessively high e-cigarette power settings by HPLC-DAD (high-performance liquid chromatography—diode array detection) carbonyl analysis. Methods: Thirteen experienced e-cigarette users evaluated whether the “dry puff flavor” was present at different power settings (10 W–25 W), recording their assessment on a 100-unit visual analog scale (VAS). They assessed e-cigarettes equipped with 1.2 Ω or 1.6 Ω coils containing menthol, vanilla or fruit-flavored e-liquids. In a machine-vaping experiment, emissions from the same liquid/coil/power setting combinations were subjected to HPLC-DAD analysis of dinitrophenol hydrazine (DNPH)-derivatized carbonyls, such as lactaldehyde and formaldehyde. A simple algorithm, based on the cutoff values for each marker, was applied to relate the dry puff flavor (as assessed by the human volunteers) to the laboratory measurements. Results: Eleven carbonyl compounds were found to agree with the human assessments. Based on the amounts of these compounds in the emissions, the dry-puff flavor did match at all combinations of e-liquids and coils examined. Dry-puff flavor was observed at different power levels with the different liquids tested. Conclusions: The described method can detect dry puff conditions and is therefore a useful tool to ensure user-relevant conditions in laboratory analyses of e-cigarette emissions. Implications: This study improves the chemical analysis of e-cigarette emissions. It offers a method to select an appropriate (i.e., user-relevant) power setting for e-cigarettes, which is a critical parameter for emission analysis and therefore important for regulatory purposes and risk assessments. Compared to the approach of using human volunteers to select appropriate power settings for different products by taste, the described method is cheaper, faster, more practical and more ethical.
Oral Health Status in Patients with Inflammatory Bowel Diseases: A Systematic Review
Authors: Nijakowski Kacper , Gruszczyński Dawid , Surdacka Anna
Published: November 2021

Inflammatory bowel diseases (IBD) are chronic disorders that affect the gastrointestinal tract, including the oral cavity. This systematic review was designed to answer the question “Is there a relationship between oral health status and inflammatory bowel diseases?”. Following the inclusion and exclusion criteria, fifteen studies were included (according to PRISMA statement guidelines). Due to their heterogeneity, only six articles about the prevalence of periodontal disease in IBD patients were included in the meta-analysis. Both Crohn’s disease (CD) and ulcerative colitis (UC) patients had an increased odds of periodontitis coincidence compared to the controls, more than 2- and 3-fold, respectively. Moreover, in most studies, patients with IBD were characterized by higher values of caries indices. In conclusion, despite the conducted systematic review, the risk of oral diseases in IBD patients cannot be clearly established due to the possible association of other factors, e.g., sociodemographic or environmental factors.
Educational Aspirations and Expectations of Adolescents in Rural China: Determinants, Mental Health, and Academic Outcomes
Authors: Chen Xiaodi , Hesketh Therese
Published: November 2021

Educational aspirations and expectations of adolescents have implications for educational and psychological outcomes. This study aimed to determine factors associated with educational aspiration–expectation discrepancies and to examine the associations between the discrepancies and psychological and academic outcomes in rural left-behind children and non-left-behind children aged 14–16 in mainland China. Cross-sectional data from a self-report survey were collected in 2020 among 606 rural students (mean age = 14.85 years) in two public middle schools in Songzi county, Hubei Province. Participants filled in questionnaires measuring their socio-demographic information, educational aspirations and expectations, academic performance, parental and friends’ aspirations, academic self-perception, academic self-regulation, depression, and self-esteem. Results showed that more than half of the participants reported that they felt they were not likely to attain the level of education to which they aspired. Parental migration, academic performance, mother’s educational aspirations for children, and close friends’ educational aspirations were the main factors associated with students’ educational aspiration–expectation discrepancies. Both left-behind children and non-left-behind children whose aspirations exceed expectations were more likely to report lower self-esteem, higher depression, lower academic self-perception, and poorer self-regulation than those without a discrepancy. These findings have implications for families, schools, and policymakers through informing the development of interventions that target positive development in rural youth.
Connexin-Based Channel Activity Is Not Specifically Altered by Hepatocarcinogenic Chemicals
Authors: Leroy Kaat , Pieters Alanah , Cooreman Axelle , Van Campenhout Raf , Cogliati Bruno , Vinken Mathieu
Published: November 2021

Connexin-based channels play key roles in cellular communication and can be affected by deleterious chemicals. In this study, the effects of various genotoxic carcinogenic compounds, non-genotoxic carcinogenic compounds and non-carcinogenic compounds on the expression and functionality of connexin-based channels, both gap junctions and connexin hemichannels, were investigated in human hepatoma HepaRG cell cultures. Expression of connexin26, connexin32, and connexin43 was evaluated by means of real-time reverse transcription quantitative polymerase chain reaction analysis, immunoblot analysis and in situ immunostaining. Gap junction functionality was assessed via a scrape loading/dye transfer assay. Opening of connexin hemichannels was monitored by measuring extracellular release of adenosine triphosphate. It was found that both genotoxic and non-genotoxic carcinogenic compounds negatively affect connexin32 expression. However, no specific effects related to chemical type were observed at gap junction or connexin hemichannel functionality level.
Chenodeoxycholic Acid Has Non-Thermogenic, Mitodynamic Anti-Obesity Effects in an In Vitro CRISPR/Cas9 Model of Bile Acid Receptor TGR5 Knockdown
Authors: Teodoro João S. , Machado Ivo F. , Castela Ana C. , Amorim João A. , Jarak Ivana , Carvalho Rui A. , Palmeira Carlos M. , Rolo Anabela P.
Published: November 2021

Bile acids (BA) have shown promising effects in animal models of obesity. However, the said effects are thought to rely on a thermogenic effect, which is questionably present in humans. A previous work has shown that the BA chenodeoxycholic acid (CDCA) can revert obesity and accelerate metabolism in animal and cell culture models. Thus, the aim of this study was to understand if this obesity reduction is indeed thermogenically-dependent. A CRISPR/Cas9 model of TGR5 (BA receptor) knockdown in 3T3-L1 adipocytes was developed to diminish thermogenic effects. Various parameters were assessed, including mitochondrial bioenergetics by Seahorse flux analysis, oxidative stress and membrane potential by fluorometry, intermediary metabolism by NMR, protein content assessment by Western Blot, gene expression by qPCR, and confocal microscopy evaluation of mitophagy. CDCA was still capable, for the most part, of reversing the harmful effects of cellular obesity, elevating mitophagy and leading to the reduction of harmed mitochondria within the cells, boosting mitochondrial activity, and thus energy consumption. In summary, CDCA has a non-thermogenic, obesity reducing capacity that hinges on a healthy mitochondrial population, explaining at least some of these effects and opening avenues of human treatment for metabolic diseases.
Ethanolamine Is a New Anti-Prion Compound
Authors: Uchiyama Keiji , Hara Hideyuki , Chida Junji , Pasiana Agriani Dini , Imamura Morikazu , Mori Tsuyoshi , Takatsuki Hanae , Atarashi Ryuichiro , Sakaguchi Suehiro
Published: November 2021

Prion diseases are a group of fatal neurodegenerative disorders caused by accumulation of proteinaceous infectious particles, or prions, which mainly consist of the abnormally folded, amyloidogenic prion protein, designated PrPSc. PrPSc is produced through conformational conversion of the cellular isoform of prion protein, PrPC, in the brain. To date, no effective therapies for prion diseases have been developed. In this study, we incidentally noticed that mouse neuroblastoma N2a cells persistently infected with 22L scrapie prions, termed N2aC24L1-3 cells, reduced PrPSc levels when cultured in advanced Dulbecco’s modified eagle medium (DMEM) but not in classic DMEM. PrPC levels remained unchanged in prion-uninfected parent N2aC24 cells cultured in advanced DMEM. These results suggest that advanced DMEM may contain an anti-prion compound(s). We then successfully identified ethanolamine in advanced DMEM has an anti-prion activity. Ethanolamine reduced PrPSc levels in N2aC24L1-3 cells, but not PrPC levels in N2aC24 cells. Also, oral administration of ethanolamine through drinking water delayed prion disease in mice intracerebrally inoculated with RML scrapie prions. These results suggest that ethanolamine could be a new anti-prion compound.
An Engineered Thermostable Laccase with Great Ability to Decolorize and Detoxify Malachite Green
Authors: Mao Guotao , Wang Kai , Wang Fangyuan , Li Hao , Zhang Hongsen , Xie Hui , Wang Zhimin , Wang Fengqin , Song Andong
Published: November 2021

Laccases can catalyze the remediation of hazardous synthetic dyes in an eco-friendly manner, and thermostable laccases are advantageous to treat high-temperature dyeing wastewater. A novel laccase from Geothermobacter hydrogeniphilus (Ghlac) was cloned and expressed in Escherichia coli. Ghlac containing 263 residues was characterized as a functional laccase of the DUF152 family. By structural and biochemical analyses, the conserved residues H78, C119, and H136 were identified to bind with one copper atom to fulfill the laccase activity. In order to make it more suitable for industrial use, Ghlac variant Mut2 with enhanced thermostability was designed. The half-lives of Mut2 at 50 °C and 60 °C were 80.6 h and 9.8 h, respectively. Mut2 was stable at pH values ranging from 4.0 to 8.0 and showed a high tolerance for organic solvents such as ethanol, acetone, and dimethyl sulfoxide. In addition, Mut2 decolorized approximately 100% of 100 mg/L of malachite green dye in 3 h at 70 °C. Furthermore, Mut2 eliminated the toxicity of malachite green to bacteria and Zea mays. In summary, the thermostable laccase Ghlac Mut2 could effectively decolorize and detoxify malachite green at high temperatures, showing great potential to remediate the dyeing wastewater.
Direct and Indirect Effect of TGFβ on Treg Transendothelial Recruitment in HCC Tissue Microenvironment
Authors: Dituri Francesco , Mancarella Serena , Serino Grazia , Chaoul Nada , Lupo Luigi Giovanni , Villa Erica , Fabregat Isabel , Giannelli Gianluigi
Published: November 2021

The balance between anti-tumor and tumor-promoting immune cells, such as CD4+ Th1 and regulatory T cells (Tregs), respectively, is assumed to dictate the progression of hepatocellular carcinoma (HCC). The transforming growth factor beta (TGFβ) markedly shapes the HCC microenvironment, regulating the activation state of multiple leukocyte subsets and driving the differentiation of cancer associated fibroblasts (CAFs). The fibrotic (desmoplastic) reaction in HCC tissue strongly depends on CAFs activity. In this study, we attempted to assess the role of TGFβ on transendothelial migration of Th1-oriented and Treg-oriented CD4+ T cells via a direct or indirect, CAF-mediated mechanisms, respectively. We found that the blockage of TGFβ receptor I-dependent signaling in Tregs resulted in impaired transendothelial migration (TEM) of these cells. Interestingly, the secretome of TGFβ-treated CAFs inhibited the TEM of Tregs but not Th1 cells, in comparison to the secretome of untreated CAFs. In addition, we found a significant inverse correlation between alpha-SMA and FoxP3 (marker of Tregs) mRNA expression in a microarray analysis involving 78 HCCs, thus suggesting that TGFβ-activated stromal cells may counteract the trafficking of Tregs into the tumor. The apparent dual behavior of TGFβ as both pro- and anti-tumorigenic cytokines may add a further level of complexity to the mechanisms that regulate the interactions among cancerous, stromal, and immune cells within HCC, as well as other solid tumors, and contribute to better manipulation of the TGFβ signaling as a therapeutic target in HCC patients.
Autophagy-Related Chemoprotection against Sorafenib in Human Hepatocarcinoma: Role of FOXO3 Upregulation and Modulation by Regorafenib
Authors: Fondevila Flavia , Méndez-Blanco Carolina , Fernández-Palanca Paula , Payo-Serafín Tania , van Pelt Jos , Verslype Chris , González-Gallego Javier , Mauriz José L.
Published: November 2021

Early acquisition of sorafenib resistance is responsible for the dismal prognosis of advanced hepatocarcinoma (HCC). Autophagy, a catabolic process involved in liver homeostasis, has been associated with chemosensitivity modulation. Forkhead box O3 (FOXO3) is a transcription factor linked to HCC pathogenesis whose role on autophagy-related sorafenib resistance remains controversial. Here, we unraveled the linkage between autophagy and sorafenib resistance in HCC, focusing on the implication of FOXO3 and its potential modulation by regorafenib. We worked with two HepG2-derived sorafenib-resistant HCC in vitro models (HepG2S1 and HepG2S3) and checked HCC patient data from the UALCAN database. Resistant cells displayed an enhanced basal autophagic flux compared to HepG2, showing higher autophagolysosome content and autophagy markers levels. Pharmacological inhibition of autophagy boosted HepG2S1 and HepG2S3 apoptosis and subG1 cells, but reduced viability, indicating the cytoprotective role of autophagy. HCC samples displayed higher FOXO3 levels, being associated with shorter survival and autophagic genes expression. Consistently, chemoresistant in vitro models showed significant FOXO3 upregulation. FOXO3 knockdown suppressed autophagy and caused resistant cell death, demonstrating that overactivation of such pro-survival autophagy during sorafenib resistance is FOXO3-dependent; a cytoprotective mechanism that the second-line drug regorafenib successfully abolished. Therefore, targeting FOXO3-mediated autophagy could significantly improve the clinical efficacy of sorafenib.
Benzyl Isothiocyanate, a Vegetable-Derived Compound, Induces Apoptosis via ROS Accumulation and DNA Damage in Canine Lymphoma and Leukemia Cells
Authors: Henklewska Marta , Pawlak Aleksandra , Li Rong-Fang , Yi Jine , Zbyryt Iwona , Obmińska-Mrukowicz Bożena
Published: November 2021

Treatment of neoplastic diseases in companion animals is one of the most important problems of modern veterinary medicine. Given the growing interest in substances of natural origin as potential anti-cancer drugs, our goal was to examine the effectiveness of benzyl isothiocyanate (BITC), a compound found in cruciferous vegetables, against canine lymphoma and leukemia. These are the one of the most common canine cancer types, and chemotherapy is the only treatment option. The study involved established cell lines originating from various hematopoietic malignancies: CLBL-1, GL-1, CLB70 and CNK-89, immortalized noncancerous cell lines: MDCK and NIH-3T3 and canine peripheral blood mononuclear cells (PBMCs). The cytotoxic activity of BITC, apoptosis induction, caspase activity and ROS generation were evaluated by flow cytometry. H2AX phosphorylation was assessed by western blot. The study showed that the compound was especially active against B lymphocyte-derived malignant cells. Their death resulted from caspase-dependent apoptosis. BITC induced ROS accumulation, and glutathione precursor N-acetyl-l-cysteine reversed the effect of the compound, thus proving the role of oxidative stress in BITC activity. In addition, exposure to the compound induced DNA damage in the tested cells. This is the first study that provides information on the activity of BITC in canine hematopoietic malignancies and suggests that the compound may be particularly useful in B-cell neoplasms treatment.
Wnt-Signaling Regulated by Glucocorticoid-Induced miRNAs
Authors: Butz Henriett , Mészáros Katalin , Likó István , Patocs Attila
Published: November 2021

Glucocorticoids (GCs) are pleiotropic hormones which regulate innumerable physiological processes. Their comprehensive effects are due to the diversity of signaling mechanism networks. MiRNAs, small, non-coding RNAs contribute to the fine tuning of signaling pathways and reciprocal regulation between GCs and miRNAs has been suggested. Our aim was to investigate the expressional change and potential function of GC mediated miRNAs. The miRNA expression profile was measured in three models: human adrenocortical adenoma vs. normal tissue, steroid-producing H295R cells and in hormonally inactive HeLa cells before and after dexamethasone treatment. The gene expression profile in 82 control and 57 GC-affected samples was evaluated in GC producing and six different GC target tissue types. Tissue-specific target prediction (TSTP) was applied to identify the most relevant miRNA−mRNA interactions. Glucocorticoid treatment resulted in cell type-dependent miRNA expression changes. However, 19.5% of the influenced signaling pathways were common in all three experiments, of which the Wnt-signaling pathway seemed to be the most affected. Transcriptome data and TSTP showed similar results, as the Wnt pathway was significantly altered in both the GC-producing adrenal gland and all investigated GC target tissue types. In different cell types, different miRNAs led to the regulation of similar pathways. Wnt signaling may be one of the most important signaling pathways affected by hypercortisolism. It is, at least in part, regulated by miRNAs that mediate the glucocorticoid effect. Our findings on GC producing and GC target tissues suggest that the alteration of Wnt signaling (together with other pathways) may be responsible for the leading symptoms observed in Cushing’s syndrome.
Surface Engineering Strategies to Enhance the In Situ Performance of Medical Devices Including Atomic Scale Engineering
Authors: Sultana Afreen , Zare Mina , Luo Hongrong , Ramakrishna Seeram
Published: November 2021

Decades of intense scientific research investigations clearly suggest that only a subset of a large number of metals, ceramics, polymers, composites, and nanomaterials are suitable as biomaterials for a growing number of biomedical devices and biomedical uses. However, biomaterials are prone to microbial infection due to Escherichia coli (E. coli), Staphylococcus aureus (S. aureus), Staphylococcus epidermidis (S. epidermidis), hepatitis, tuberculosis, human immunodeficiency virus (HIV), and many more. Hence, a range of surface engineering strategies are devised in order to achieve desired biocompatibility and antimicrobial performance in situ. Surface engineering strategies are a group of techniques that alter or modify the surface properties of the material in order to obtain a product with desired functionalities. There are two categories of surface engineering methods: conventional surface engineering methods (such as coating, bioactive coating, plasma spray coating, hydrothermal, lithography, shot peening, and electrophoretic deposition) and emerging surface engineering methods (laser treatment, robot laser treatment, electrospinning, electrospray, additive manufacturing, and radio frequency magnetron sputtering technique). Atomic-scale engineering, such as chemical vapor deposition, atomic layer etching, plasma immersion ion deposition, and atomic layer deposition, is a subsection of emerging technology that has demonstrated improved control and flexibility at finer length scales than compared to the conventional methods. With the advancements in technologies and the demand for even better control of biomaterial surfaces, research efforts in recent years are aimed at the atomic scale and molecular scale while incorporating functional agents in order to elicit optimal in situ performance. The functional agents include synthetic materials (monolithic ZnO, quaternary ammonium salts, silver nano-clusters, titanium dioxide, and graphene) and natural materials (chitosan, totarol, botanical extracts, and nisin). This review highlights the various strategies of surface engineering of biomaterial including their functional mechanism, applications, and shortcomings. Additionally, this review article emphasizes atomic scale engineering of biomaterials for fabricating antimicrobial biomaterials and explores their challenges.
Time-Dependent Image Restoration of Low-SNR Live-Cell Ca2 Fluorescence Microscopy Data
Authors: Woelk Lena-Marie , Kannabiran  Sukanya A. , Brock  Valerie J. , Gee  Christine E. , Lohr  Christian , Guse  Andreas H. , Diercks  Björn-Philipp , Werner René
Published: November 2021

Live-cell Ca2+ fluorescence microscopy is a cornerstone of cellular signaling analysis and imaging. The demand for high spatial and temporal imaging resolution is, however, intrinsically linked to a low signal-to-noise ratio (SNR) of the acquired spatio-temporal image data, which impedes on the subsequent image analysis. Advanced deconvolution and image restoration algorithms can partly mitigate the corresponding problems but are usually defined only for static images. Frame-by-frame application to spatio-temporal image data neglects inter-frame contextual relationships and temporal consistency of the imaged biological processes. Here, we propose a variational approach to time-dependent image restoration built on entropy-based regularization specifically suited to process low- and lowest-SNR fluorescence microscopy data. The advantage of the presented approach is demonstrated by means of four datasets: synthetic data for in-depth evaluation of the algorithm behavior; two datasets acquired for analysis of initial Ca2+ microdomains in T-cells; finally, to illustrate the transferability of the methodical concept to different applications, one dataset depicting spontaneous Ca2+ signaling in jGCaMP7b-expressing astrocytes. To foster re-use and reproducibility, the source code is made publicly available.
Anabolic Steroids-Driven Regulation of Porcine Ovarian Putative Stem Cells Favors the Onset of Their Neoplastic Transformation
Authors: Gorczyca Gabriela , Wartalski Kamil , Wiater Jerzy , Samiec Marcin , Tabarowski Zbigniew , Duda Małgorzata
Published: November 2021

Nandrolone (Ndn) and boldenone (Bdn), the synthetic testosterone analogues with strong anabolic effects, despite being recognized as potentially carcinogenic compounds, are commonly abused by athletes and bodybuilders, which includes women, worldwide. This study tested the hypothesis that different doses of Ndn and Bdn can initiate neoplastic transformation of porcine ovarian putative stem cells (poPSCs). Immunomagnetically isolated poPSCs were expanded ex vivo in the presence of Ndn or Bdn, for 7 and 14 days. Results show that pharmacological doses of both Ndn and Bdn, already after 7 days of poPSCs culture, caused a significant increase of selected, stemness-related markers of cancer cells: CD44 and CD133. Notably, Ndn also negatively affected poPSCs growth not only by suppressing their proliferation and mitochondrial respiration but also by inducing apoptosis. This observation shows, for the first time, that chronic exposure to Ndn or Bdn represents a precondition that might enhance risk of poPSCs neoplastic transformation. These studies carried out to accomplish detailed molecular characterization of the ex vivo expanded poPSCs and their potentially cancerous derivatives (PCDs) might be helpful to determine their suitability as nuclear donor cells (NDCs) for further investigations focused on cloning by somatic cell nuclear transfer (SCNT). Such investigations might also be indispensable to estimate the capabilities of nuclear genomes inherited from poPSCs and their PCDs to be epigenetically reprogrammed (dedifferentiated) in cloned pig embryos generated by SCNT. This might open up new possibilities for biomedical research aimed at more comprehensively recognizing genetic and epigenetic mechanisms underlying not only tumorigenesis but also reversal/retardation of pro-tumorigenic intracellular events.
Fertility-Sparing Approach in Women Affected by Stage I and Low-Grade Endometrial Carcinoma: An Updated Overview
Authors: Gullo Giuseppe , Etrusco Andrea , Cucinella Gaspare , Perino Antonino , Chiantera Vito , Laganà Antonio Simone , Tomaiuolo Rossella , Vitagliano Amerigo , Giampaolino Pierluigi , Noventa Marco , Andrisani Alessandra , Buzzaccarini Giovanni
Published: November 2021

Endometrial cancer (EC) is a deleterious condition which strongly affects a woman’s quality of life. Although aggressive interventions should be considered to treat high-grade EC, a conservative approach should be taken into consideration for women wishing to conceive. In this scenario, we present an overview about the EC fertility-sparing approach state of art. Type I EC at low stage is the only histological type which can be addressed with a fertility-sparing approach. Moreover, no myometrium and/or adnexal invasion should be seen, and lymph-vascular space should not be involved. Regarding the pharmaceutical target, progestins, in particular medroxyprogesterone acetate (MPA) or megestrol acetate (MA), are the most employed agent in conservative treatment of early-stage EC. The metformin usage and hysteroscopic assessment is still under debate, despite promising results. Particularly strict and imperious attention should be given to the follow-up and psychological wellbeing of women, especially because of the double detrimental impairment: both EC and EC-related infertility consequences.
Periodontitis and Gestational Diabetes Mellitus: A Potential Inflammatory Vicious Cycle
Authors: Bendek María José , Canedo-Marroquín Gisela , Realini Ornella , Retamal Ignacio N. , Hernández Marcela , Hoare Anilei , Busso Dolores , Monteiro Lara J. , Illanes Sebastián E. , Chaparro Alejandra
Published: November 2021

Periodontitis is a chronic inflammatory immune disease associated with a dysbiotic state, influenced by keystone bacterial species responsible for disrupting the periodontal tissue homeostasis. Furthermore, the severity of periodontitis is determined by the interaction between the immune cell response in front of periodontitis-associated species, which leads to the destruction of supporting periodontal tissues and tooth loss in a susceptible host. The persistent bacterial challenge induces modifications in the permeability and ulceration of the sulcular epithelium, which facilitates the systemic translocation of periodontitis-associated bacteria into distant tissues and organs. This stimulates the secretion of pro-inflammatory molecules and a chronic activation of immune cells, contributing to a systemic pro-inflammatory status that has been linked with a higher risk of several systemic diseases, such as type 2 diabetes mellitus (T2DM) and gestational diabetes mellitus (GDM). Although periodontitis and GDM share the common feature of systemic inflammation, the molecular mechanistic link of this association has not been completely clarified. This review aims to examine the potential biological mechanisms involved in the association between periodontitis and GDM, highlighting the contribution of both diseases to systemic inflammation and the role of new molecular participants, such as extracellular vesicles and non-coding RNAs, which could act as novel molecular intercellular linkers between periodontal and placental tissues.
CRISPRi-Guided Metabolic Flux Engineering for Enhanced Protopanaxadiol Production in Saccharomyces cerevisiae
Authors: Lim Soo-Hwan , Baek Jong-In , Jeon Byeong-Min , Seo Jung-Woo , Kim Min-Sung , Byun Ji-Young , Park Soo-Hoon , Kim Su-Jin , Lee Ju-Young , Lee Jun-Hyoung , Kim Sun-Chang
Published: November 2021

Protopanaxadiol (PPD), an aglycon found in several dammarene-type ginsenosides, has high potency as a pharmaceutical. Nevertheless, application of these ginsenosides has been limited because of the high production cost due to the rare content of PPD in Panax ginseng and a long cultivation time (4–6 years). For the biological mass production of the PPD, de novo biosynthetic pathways for PPD were introduced in Saccharomyces cerevisiae and the metabolic flux toward the target molecule was restructured to avoid competition for carbon sources between native metabolic pathways and de novo biosynthetic pathways producing PPD in S. cerevisiae. Here, we report a CRISPRi (clustered regularly interspaced short palindromic repeats interference)-based customized metabolic flux system which downregulates the lanosterol (a competing metabolite of dammarenediol-II (DD-II)) synthase in S. cerevisiae. With the CRISPRi-mediated suppression of lanosterol synthase and diversion of lanosterol to DD-II and PPD in S. cerevisiae, we increased PPD production 14.4-fold in shake-flask fermentation and 5.7-fold in a long-term batch-fed fermentation.
Development of In Vitro and In Vivo Evaluation Systems for Vitamin D Derivatives and Their Application to Drug Discovery
Authors: Yasuda Kaori , Nishikawa Miyu , Mano Hiroki , Takano Masashi , Kittaka Atsushi , Ikushiro Shinichi , Sakaki Toshiyuki
Published: November 2021

We have developed an in vitro system to easily examine the affinity for vitamin D receptor (VDR) and CYP24A1-mediated metabolism as two methods of assessing vitamin D derivatives. Vitamin D derivatives with high VDR affinity and resistance to CYP24A1-mediated metabolism could be good therapeutic agents. This system can effectively select vitamin D derivatives with these useful properties. We have also developed an in vivo system including a Cyp27b1-gene-deficient rat (a type I rickets model), a Vdr-gene-deficient rat (a type II rickets model), and a rat with a mutant Vdr (R270L) (another type II rickets model) using a genome editing method. For Cyp27b1-gene-deficient and Vdr mutant (R270L) rats, amelioration of rickets symptoms can be used as an index of the efficacy of vitamin D derivatives. Vdr-gene-deficient rats can be used to assess the activities of vitamin D derivatives specialized for actions not mediated by VDR. One of our original vitamin D derivatives, which displays high affinity VDR binding and resistance to CYP24A1-dependent metabolism, has shown good therapeutic effects in Vdr (R270L) rats, although further analysis is needed.
Long-Chain and Very Long-Chain Ceramides Mediate Doxorubicin-Induced Toxicity and Fibrosis
Authors: Kretzschmar Tom , Bekhite Mohamed M. , Wu Jasmine M. F. , Haase Daniela , Förster Martin , Müller Tina , Nietzsche Sandor , Westermann Martin , Franz Marcus , Gräler Markus H. , Schulze P. Christian
Published: November 2021

Doxorubicin (Dox) is a chemotherapeutic agent with cardiotoxicity associated with profibrotic effects. Dox increases ceramide levels with pro-inflammatory effects, cell death, and fibrosis. The purpose of our study was to identify the underlying ceramide signaling pathways. We aimed to characterize the downstream effects on cell survival, metabolism, and fibrosis. Human fibroblasts (hFSF) were treated with 0.7 µM of Dox or transgenically overexpressed ceramide synthase 2 (FLAG-CerS2). Furthermore, cells were pre-treated with MitoTempo (MT) (2 h, 20 µM) or Fumonisin B1 (FuB) (4 h, 100 µM). Protein expression was measured by Western blot or immunofluorescence (IF). Ceramide levels were determined with mass spectroscopy (MS). Visualizations were conducted using laser scanning microscopy (LSM) or electron microscopy. Mitochondrial activity was measured using seahorse analysis. Dox and CerS2 overexpression increased CerS2 protein expression. Coherently, ceramides were elevated with the highest peak for C24:0. Ceramide- induced mitochondrial ROS production was reduced with MT or FuB preincubation. Mitochondrial homeostasis was reduced and accompanied by reduced ATP production. Our data show that the increase in pro-inflammatory ceramides is an essential contributor to Dox side-effects. The accumulation of ceramides resulted in a lipotoxic shift and subsequently mitochondrial structural and functional damage, which was partially reversible following inhibition of ceramide synthesis.
Recent Advances in Diabetic Kidney Diseases: From Kidney Injury to Kidney Fibrosis
Authors: Hung Peir-Haur , Hsu Yung-Chien , Chen Tsung-Hsien , Lin Chun-Liang
Published: November 2021

Diabetic kidney disease (DKD) is the leading cause of chronic kidney disease and end-stage renal disease. The natural history of DKD includes glomerular hyperfiltration, progressive albuminuria, declining estimated glomerular filtration rate, and, ultimately, kidney failure. It is known that DKD is associated with metabolic changes caused by hyperglycemia, resulting in glomerular hypertrophy, glomerulosclerosis, and tubulointerstitial inflammation and fibrosis. Hyperglycemia is also known to cause programmed epigenetic modification. However, the detailed mechanisms involved in the onset and progression of DKD remain elusive. In this review, we discuss recent advances regarding the pathogenic mechanisms involved in DKD.
Bio-Performance of Hydrothermally and Plasma-Treated Titanium: The New Generation of Vascular Stents
Authors: Benčina Metka , Rawat Niharika , Lakota Katja , Sodin-Šemrl Snežna , Iglič Aleš , Junkar Ita
Published: November 2021

The research presented herein follows an urgent global need for the development of novel surface engineering techniques that would allow the fabrication of next-generation cardiovascular stents, which would drastically reduce cardiovascular diseases (CVD). The combination of hydrothermal treatment (HT) and treatment with highly reactive oxygen plasma (P) allowed for the formation of an oxygen-rich nanostructured surface. The morphology, surface roughness, chemical composition and wettability of the newly prepared oxide layer on the Ti substrate were characterized by scanning electron microscopy (SEM) with energy-dispersive X-ray analysis (EDX), atomic force microscopy (AFM), X-ray photoelectron spectroscopy (XPS) and water contact angle (WCA) analysis. The alteration of surface characteristics influenced the material’s bio-performance; platelet aggregation and activation was reduced on surfaces treated by hydrothermal treatment, as well as after plasma treatment. Moreover, it was shown that surfaces treated by both treatment procedures (HT and P) promoted the adhesion and proliferation of vascular endothelial cells, while at the same time inhibiting the adhesion and proliferation of vascular smooth muscle cells. The combination of both techniques presents a novel approach for the fabrication of vascular implants, with superior characteristics.
GntR-like SCO3932 Protein Provides a Link between Actinomycete Integrative and Conjugative Elements and Secondary Metabolism
Authors: Pawlik Krzysztof J. , Zelkowski Mateusz , Biernacki Mateusz , Litwinska Katarzyna , Jaworski Pawel , Kotowska Magdalena
Published: November 2021

Streptomyces bacteria produce a plethora of secondary metabolites including the majority of medically important antibiotics. The onset of secondary metabolism is correlated with morphological differentiation and controlled by a complex regulatory network involving numerous regulatory proteins. Control over these pathways at the molecular level has a medical and industrial importance. Here we describe a GntR-like DNA binding transcription factor SCO3932, encoded within an actinomycete integrative and conjugative element, which is involved in the secondary metabolite biosynthesis regulation. Affinity chromatography, electrophoresis mobility shift assay, footprinting and chromatin immunoprecipitation experiments revealed, both in vitro and in vivo, SCO3932 binding capability to its own promoter region shared with the neighboring gene SCO3933, as well as promoters of polyketide metabolite genes, such as cpkD, a coelimycin biosynthetic gene, and actII-orf4—an activator of actinorhodin biosynthesis. Increased activity of SCO3932 target promoters, as a result of SCO3932 overproduction, indicates an activatory role of this protein in Streptomyces coelicolor A3(2) metabolite synthesis pathways.
Early-Life Origins of Metabolic Syndrome: Mechanisms and Preventive Aspects
Authors: Hsu Chien-Ning , Hou Chih-Yao , Hsu Wei-Hsuan , Tain You-Lin
Published: November 2021

One of the leading global public-health burdens is metabolic syndrome (MetS), despite the many advances in pharmacotherapies. MetS, now known as “developmental origins of health and disease” (DOHaD), can have its origins in early life. Offspring MetS can be programmed by various adverse early-life conditions, such as nutrition imbalance, maternal conditions or diseases, maternal chemical exposure, and medication use. Conversely, early interventions have shown potential to revoke programming processes to prevent MetS of developmental origins, namely reprogramming. In this review, we summarize what is currently known about adverse environmental insults implicated in MetS of developmental origins, including the fundamental underlying mechanisms. We also describe animal models that have been developed to study the developmental programming of MetS. This review extends previous research reviews by addressing implementation of reprogramming strategies to prevent the programming of MetS. These mechanism-targeted strategies include antioxidants, melatonin, resveratrol, probiotics/prebiotics, and amino acids. Much work remains to be accomplished to determine the insults that could induce MetS, to identify the mechanisms behind MetS programming, and to develop potential reprogramming strategies for clinical translation.
Editorial for the Special Issue “Molecular Bases of Senescence”
Authors: Fanò-Illic Giorgio , Fulle Stefania , Mecocci Patrizia
Published: November 2021

The increasing life expectancy of populations worldwide represents the most evident success of the last century thanks to varying interacting social and medical achievements [...]
Differential Clearance of Aβ Species from the Brain by Brain Lymphatic Endothelial Cells in Zebrafish
Authors: Jeong Yun-Mi , Lee Jae-Geun , Cho Hyun-Ju , Lee Wang Sik , Jeong Jinyoung , Lee Jeong-Soo
Published: November 2021

The failure of amyloid beta (Aβ) clearance is a major cause of Alzheimer’s disease, and the brain lymphatic systems play a crucial role in clearing toxic proteins. Recently, brain lymphatic endothelial cells (BLECs), a non-lumenized lymphatic cell in the vertebrate brain, was identified, but Aβ clearance via this novel cell is not fully understood. We established an in vivo zebrafish model using fluorescently labeled Aβ42 to investigate the role of BLECs in Aβ clearance. We discovered the efficient clearance of monomeric Aβ42 (mAβ42) compared to oligomeric Aβ42 (oAβ42), which was illustrated by the selective uptake of mAβ42 by BLECs and peripheral transport. The genetic depletion, pharmacological inhibition via the blocking of the mannose receptor, or the laser ablation of BLECs resulted in the defective clearance of mAβ42. The treatment with an Aβ disaggregating agent facilitated the internalization of oAβ42 into BLECs and improved the peripheral transport. Our findings reveal a new role of BLECs in the differential clearance of mAβ42 from the brain and provide a novel therapeutic strategy based on promoting Aβ clearance.
Melatonin-Mediated Colonic Microbiota Metabolite Butyrate Prevents Acute Sleep Deprivation-Induced Colitis in Mice
Authors: Gao Ting , Wang Zixu , Dong Yulan , Cao Jing , Chen Yaoxing
Published: November 2021

Radical cure colitis is a severe public health threat worldwide. Our previous studies have confirmed that melatonin can effectively improve gut microbiota disorder and mucosal injury caused by sleep deprivation (SD). The present study further explored the mechanism whereby exogenous melatonin prevented SD-induced colitis. 16S rRNA high-throughput sequencing and metabolomics analysis were used to explore the correlation between SD-induced colitis and intestinal microbiota and metabolite composition in mice. Fecal microbiota transplantation (FMT) and melatonin or butyrate supplementation tests verified the core role of gut microbiota in melatonin-alleviating SD-induced colitis. Further, in vitro tests studied the modulatory mechanism of metabolite butyrate. The results demonstrated that SD leads to reductions in plasma melatonin levels and colonic Card9 expression and consequent occurrence of colitis and gut microbiota disorder, especially the downregulation of Faecalibacterium and butyrate levels. The FMT from SD-mice to normal mice could restore SD-like colitis, while butyrate supplementation to SD-mice inhibited the occurrence of colitis, but with no change in the plasma melatonin level in both treatments. However, melatonin supplementation reversed all inductions in SD-mice. In intestinal epithelial cells, the inflammatory ameliorative effect of butyrate was blocked with pretreatments of HDAC3 agonist and HIF-1α antagonist but was mimicked by GSK-3β and p-P65 antagonists. Therefore, the administration of MLT may be a better therapy for SD-induced colitis relative to butyrate. A feasible mechanism would involve that melatonin up-regulated the Faecalibacterium population and production of its metabolite butyrate and MCT1 expression and inhibited HDAC3 in the colon, which would allow p-GSK-3β/β-catenin/HIF-1α activation and NF-κB/NLRP3 suppression to up-regulate Card9 expression and suppress inflammation response.