bluuebunny/crossref_metadata_2025 huggingface dataset:
Towards the Improved Accuracy of Hepatitis E Diagnosis in Vulnerable and Target Groups: A Global Perspective on the Current State of Knowledge and the Implications for Practice
Authors: Talapko Jasminka , Meštrović Tomislav , Pustijanac Emina , Škrlec Ivana
Published: January 2021

The hepatitis E virus (HEV) is a positive single-stranded, icosahedral, quasi-enveloped RNA virus in the genus Orthohepevirus of the family Hepeviridae. Orthohepevirus A is the most numerous species of the genus Orthohepevirus and consists of eight different HEV genotypes that can cause infection in humans. HEV is a pathogen transmitted via the fecal–oral route, most commonly by consuming fecally contaminated water. A particular danger is the HEV-1 genotype, which poses a very high risk of vertical transmission from the mother to the fetus. Several outbreaks caused by this genotype have been reported, resulting in many premature births, abortions, and also neonatal and maternal deaths. Genotype 3 is more prevalent in Europe; however, due to the openness of the market, i.e., trade-in animals which represent a natural reservoir of HEV (such as pigs), there is a possibility of spreading HEV infections outside endemic areas. This problem is indeed global and requires increased hygiene measures in endemic areas, which entails special care for pregnant women in both endemic and non-endemic regions. As already highlighted, pregnant women could have significant health consequences due to the untimely diagnosis of HEV infection; hence, this is a population that should be targeted with a specific combination of testing approaches to ensure optimal specificity and sensitivity. Until we advance from predominantly supportive treatment in pregnancy and appraise the safety and efficacy of a HEV vaccine in this population, such screening approaches represent the mainstay of our public health endeavors.
Laser Photobiomodulation (PBM)—A Possible New Frontier for the Treatment of Oral Cancer: A Review of In Vitro and In Vivo Studies
Authors: Del Vecchio Alessandro , Tenore Gianluca , Luzi Maria Clotilde , Palaia Gaspare , Mohsen Ahmed , Pergolini Daniele , Romeo Umberto
Published: January 2021

The treatment of oral squamous cell carcinoma (OSCC) is particularly complex due to its aggressive behavior, location, the patient’s age, and its spread at diagnosis. In recent years, photobiomodulation (PBM) has been introduced in different medical fields; however, its application, in patients suffering from OSCC for palliative support or to induce analgesia, has been hotly debated due to the possibility that the cell growth stimuli induced by PBM could lead to a worsening of the lesions. The aim of this study is to review the literature to observe the available data investigating the effect of PBM on cancer cells in vitro and in vivo. A review was conducted on the PubMed and Scopus databases. A total of twelve studies met the inclusion criteria and were therefore included for quality assessment and data extraction. The analysis showed that the clinical use of PBM is still only partially understood and is, therefore, controversial. Some authors stated that it could be contraindicated for clinical use in patients suffering from SCC, while others noted that it could have beneficial effects. According to the data that emerged from this review, it is possible to hypothesize that there are possibilities for PBM to play a beneficial role in treating cancer patients, but further evidence about its clinical efficacy and the identification of protocols and correct dosages is still needed.
Conceptual and Mathematical Modeling of a Coastal Aquifer in Eastern Delta of R. Nestos (N. Greece)
Authors: Gkiougkis Ioannis , Pouliaris Christos , Pliakas Fotios-Konstantinos , Diamantis Ioannis , Kallioras Andreas
Published: January 2021

In this paper, the development of the conceptual and groundwater flow model for the coastal aquifer system of the alluvial plain of River Nestos (N. Greece), that suffers from seawater intrusion due to over-pumping for irrigation, is analyzed. The study area is a typical semi-arid hydrogeologic environment, composed of a multi-layer granular aquifers that covers the eastern coastal delta system of R. Nestos. This study demonstrates the results of a series of field measurements (such as geophysical surveys, hydrochemical and isotopical measurements, hydro-meteorological data, land use, irrigation schemes) that were conducted during the period 2009 to 2014. The synthesis of the above resulted in the development of the conceptual model for this aquifer system, that formed the basis for the application of the mathematical model for simulating groundwater flow. The mathematical modeling was achieved using the finite difference method after the application of the USGS code MODFLOW-2005.
The Cardiometabolic Health Benefits of Sauna Exposure in Individuals with High-Stress Occupations. A Mechanistic Review
Authors: Henderson Kaemmer N. , Killen Lauren G. , O’Neal Eric K. , Waldman Hunter S.
Published: January 2021

Components of the metabolic syndrome (i.e., hypertension, insulin resistance, obesity, atherosclerosis) are a leading cause of death in the United States and result in low-grade chronic inflammation, excessive oxidative stress, and the eventual development of cardiometabolic diseases (CMD). High-stress occupations (HSO: firefighters, police, military personnel, first responders, etc.) increase the risk of developing CMD because they expose individuals to chronic and multiple stressors (i.e., sleep deprivation, poor nutrition habits, lack of physical activity, psychological stress). Interestingly, heat exposure and, more specifically, sauna bathing have been shown to improve multiple markers of CMD, potentially acting as hormetic stressors, at the cellular level and in the whole organism. Therefore, sauna bathing might be a practical and alternative intervention for disease prevention for individuals with HSO. The purpose of this review is to detail the mechanisms and pathways involved in the response to both acute and chronic sauna bathing and collectively present sauna bathing as a potential treatment, in addition to current standard of care, for mitigating CMD to both clinicians and individuals serving in HSO.
Nonmedical Use of Benzodiazepines among Immigrant and Native-Born Adolescents in Spain: National Trends and Related Factors
Authors: Carrasco-Garrido Pilar , Díaz Rodríguez Dania Rocío , Jiménez-Trujillo Isabel , Hernández-Barrera Valentín , Lima Florencio Lidiane , Palacios-Ceña Domingo
Published: January 2021

Background: the nonmedical use of prescribed medications among adolescents has increased significantly in recent years. We aimed to identify the patterns of benzodiazepine nonmedical use and its evolution during the decade 2006–2016 among immigrant and native-born adolescent populations. Methods: we used individualized secondary data retrieved from the 2006–2016 Spanish State Survey on Drug Use in Secondary Education (ESTUDES) of the school-aged population. Using logistic multivariate regression models, we estimated the independent effect of each of these variables on nonmedical use. Two models were generated: one for immigrant adolescents and one for native-born adolescents. Results: during the decade 2006–2016, 2.81% of native-born and 3.36% of immigrant adolescent students made nonmedical use of benzodiazepines. Gender and socioeconomic status were found to be related to the nonmedical use of benzodiazepines. Consumption of illegal psychoactive substances, other than marijuana, was the variable of greatest value (aOR = 6.00, 95% CI 3.89–9.27). Perceived risks and drug availability were found to be predictors for the nonmedical use of benzodiazepines in both immigrant and native-born adolescents. Conclusion: in Spain, patterns of benzodiazepine nonmedical use among immigrant and native-born adolescents are similar. The results of this study refute certain stereotypes related to consumption of substances among immigrant adolescents, identifying them as a risk group.
Traffic Crash Characteristics in Shenzhen, China from 2014 to 2016
Authors: Li Guofa , Liao Yuan , Guo Qiangqiang , Shen Caixiong , Lai Weijian
Published: January 2021

Road traffic crashes cause fatalities and injuries of both drivers/passengers in vehicles and pedestrians outside, thus challenge public health especially in big cities in developing countries like China. Previous efforts mainly focus on a specific crash type or causation to examine the crash characteristics in China while lacking the characteristics of various crash types, factors, and the interplay between them. This study investigated the crash characteristics in Shenzhen, one of the biggest four cities in China, based on the police-reported crashes from 2014 to 2016. The descriptive characteristics were reported in detail with respect to each of the crash attributes. Based on the recorded crash locations, the land-use pattern was obtained as one of the attributes for each crash. Then, the relationship between the attributes in motor-vehicle-involved crashes was examined using the Bayesian network analysis. We revealed the distinct crash characteristics observed between the examined levels of each attribute, as well the interplay between the attributes. This study provides an insight into the crash characteristics in Shenzhen, which would help understand the driving behavior of Chinese drivers, identify the traffic safety problems, guide the research focuses on advanced driver assistance systems (ADASs) and traffic management countermeasures in China.
Environmental Impact Assessment in the Former Mining Area of Regoufe (Arouca, Portugal): Contributions to Future Remediation Measures
Authors: Durães Nuno , Portela Luís , Sousa Sara , Patinha Carla , da Silva Eduardo Ferreira
Published: January 2021

The W-Sn Regoufe mine, closed since the 1970s, was once intensively exploited for tungsten concentrates. Throughout its activity, considerable amounts of arsenopyrite-rich mine wastes were produced and, to this day, are still exposed to weathering conditions. Thus, this work aims at assessing soil contamination, using a combination of chemical, physicochemical and mineralogical analyses and sequential selective chemical extraction of the main potentially toxic elements (PTEs) in topsoils. Results show that Regoufe soils are enriched in most of the PTEs associated with the ore assemblage, but As and Cd contents far outstrip both international and national guidelines. The estimated contamination factor reveals that 67% of soil samples are classified as highly to ultra-highly contaminated. Similar distribution patterns, with the main focus around the unsealed mine adits, are observed when spatially projecting the modified degree of contamination (mCd) and arsenic contents. Fe-oxyhydroxides and organic matter demonstrate to have a preponderant role in the retention of Cd and As. In fact, despite the high PTE contents in soils, local surface waters are characterised by low metal(loid) contents and nearly neutral pH, with PTE concentrations below national thresholds for irrigation waters.
Associations of Perfluoroalkyl Substances with Prevalence of Metabolic Syndrome in Highly Exposed Young Adult Community Residents—A Cross-Sectional Study in Veneto Region, Italy
Authors: Zare Jeddi Maryam , Dalla Zuanna Teresa , Barbieri Giulia , Fabricio Aline S. C. , Daprà Francesca , Fletcher Tony , Russo Francesca , Pitter Gisella , Canova Cristina
Published: January 2021

Background: Studies on the association between perfluoroalkyl substances (PFAS) and metabolic syndrome (MetS) are limited, and results are inconsistent. We aimed to examine the associations between PFAS serum levels and the prevalence of MetS among highly exposed young adults (ages 20–39) residents of a large area of the Veneto Region (North-Eastern Italy) primarily stemming from PFAS water contamination before September 2013. A total of 15,876 eligible young adult residents living in the investigated municipalities were enrolled in the study from January 2017 to July 2019. Methods: MetS was defined by using a modified harmonized definition requiring the presence of 3 of the following: obesity (body mass index ≥30), elevated triglyceride (TG), reduced high-density lipoprotein cholesterol, elevated blood pressure, and hemoglobin A1c ≥ 6.1% or self-reported diabetes mellitus or drug treatment for hyperglycemia. Multivariable generalized additive models were performed to identify the associations between four serum PFAS, including perfluorooctane sulfonic acid (PFOS), perfluorooctanoic acid (PFOA), perfluorohexane sulfonic acid (PFHxS), and perfluorononanoic acid (PFNA), and risk of MetS controlling for potential confounders. Results: A total of 1282 participants (8.1%) met the criteria of MetS with a higher prevalence among men. PFOA, PFHxS, and PFNA were not associated with the risk of MetS, whereas PFOS showed a consistent protective effect against the risk of MetS (OR 0.76, (95% CI: 0.69, 0.85) per ln-PFOS). However, we found statistically significant positive associations between PFAS serum levels and individual components of MetS, mainly elevated blood pressure and elevated TG. Conclusion: Our results did not support a consistent association between PFAS and MetS and conflicting findings were observed for individual components of MetS.
Effects of HIIT and MIIT Suspension Training Programs on Sleep Quality and Fatigue in Older Adults: Randomized Controlled Clinical Trial
Authors: Jiménez-García José Daniel , Hita-Contreras Fidel , de la Torre-Cruz Manuel Jesús , Aibar-Almazán Agustín , Achalandabaso-Ochoa Alexander , Fábrega-Cuadros Raquel , Martínez-Amat Antonio
Published: January 2021

Poor sleep quality lessens general health quality and is related to physical and mental problems. Moreover, fatigue is one of the foremost common complaints in medical care and plays a role in the decreasing quality of life of the older population. For these reasons, the objective of this study was to examine the effect of high- and moderate-intensity interval training programs (HIIT vs. MIIT)—both consisting of twelve weeks of TRX training—on the sleep quality and fatigue levels of the elderly. A randomized controlled clinical trial (NCT03404830) was conducted. A total of 82 subjects were randomized to either a HIIT group (n = 28) that performed a main squat activity with a suspension system, comprising four four-minute intervals between 90–95% of the maximum heart rate (HR), an MIIT group (n = 27) with an intensity of 70% of the maximum HR, and a control group (CG) (n = 27) that continued their daily lifestyle. The two exercise groups trained twice a week for 12 weeks, with each session lasting 45 min. Sleep quality was measured using the Pittsburgh sleep quality index (PSQI), and fatigue was assessed using the fatigue severity scale (FSS). Outcomes were measured before the intervention and after the intervention period. Post-intervention sleep quality measurements revealed a statistically significant interaction regarding group × time (p < 0.005) and fatigue (p = 0.002). Specifically, fatigue decreased in the HIIT group between both measurement moments (p = 0.003). In addition, differences were obtained in the post-intervention measure between the HIIT and MIIT groups (p = 0.013) and HIIT and control (p = 0.029). Our analysis indicates that a population of the elderly showed improvements in sleep quality and fatigue after performing a high-intensity intervention using suspension training (TRX), with markedly better results in the HIIT group.
Regulating and Cultural Ecosystem Services of Urban Green Infrastructure in the Nordic Countries: A Systematic Review
Authors: Amorim Jorge H. , Engardt Magnuz , Johansson Christer , Ribeiro Isabel , Sannebro Magnus
Published: January 2021

In the Nordic countries (Denmark, Finland, Iceland, Norway and Sweden), the Urban Green Infrastructure (UGI) has been traditionally targeted at reducing flood risk. However, other Ecosystem Services (ES) became increasingly relevant in response to the challenges of urbanization and climate change. In total, 90 scientific articles addressing ES considered crucial contributions to the quality of life in cities are reviewed. These are classified as (1) regulating ES that minimize hazards such as heat, floods, air pollution and noise, and (2) cultural ES that promote well-being and health. We conclude that the planning and design of UGI should balance both the provision of ES and their side effects and disservices, aspects that seem to have been only marginally investigated. Climate-sensitive planning practices are critical to guarantee that seasonal climate variability is accounted for at high-latitude regions. Nevertheless, diverging and seemingly inconsistent findings, together with gaps in the understanding of long-term effects, create obstacles for practitioners. Additionally, the limited involvement of end users points to a need of better engagement and communication, which in overall call for more collaborative research. Close relationships and interactions among different ES provided by urban greenery were found, yet few studies attempted an integrated evaluation. We argue that promoting interdisciplinary studies is fundamental to attain a holistic understanding of how plant traits affect the resulting ES; of the synergies between biophysical, physiological and psychological processes; and of the potential disservices of UGI, specifically in Nordic cities.
Access to Healthcare Following Serious Injury: Perspectives of Allied Health Professionals in Urban and Regional Settings
Authors: Keeves Jemma , Braaf Sandra C. , Ekegren Christina L. , Beck Ben , Gabbe Belinda J.
Published: January 2021

Barriers to accessing healthcare exist following serious injury. These issues are not well understood and may have dire consequences for healthcare utilisation and patients’ long-term recovery. The aim of this qualitative study was to explore factors perceived by allied health professionals to affect access to healthcare beyond hospital discharge for people with serious injuries in urban and regional Victoria, Australia. Twenty-five semi-structured interviews were conducted with community-based allied health professionals involved in post-discharge care for people following serious injury across different urban and regional areas. Interview transcripts were analysed using thematic analysis. Many allied health professionals perceived that complex funding systems and health services restrict access in both urban and regional areas. Limited availability of necessary health professionals was consistently reported, which particularly restricted access to mental healthcare. Access to healthcare was also felt to be hindered by a reliance on others for transportation, costs, emotional stress and often lengthy time of travel. Across urban and regional areas, a number of factors limit access to healthcare. Better understanding of health service delivery models and areas for change, including the use of technology and telehealth, may improve equitable access to healthcare.
The Paternal Experience of Fear of Childbirth: An Integrative Review
Authors: Moran Emma , Bradshaw Carmel , Tuohy Teresa , Noonan Maria
Published: January 2021

Background: It is estimated that approximately 13% of expectant fathers experience a pathological and debilitating fear of childbirth. Objective: The aim of this integrative review was to examine and synthesise the current body of research relating to paternal experience of fear of childbirth. Methods: A systematic literature search of five databases—CINAHL, Cochrane Library, MEDLINE, PsycArticles and PsycInfo—identified seventeen papers. Methodological quality of studies was assessed using the Crowe Critical Appraisal Tool. Results: Thematic data analysis identified three themes: the focus of fathers’ childbirth-related fears, the impact of fear of childbirth on health and wellbeing, and fear of childbirth as a private burden. Discussion: Fear of childbirth is a significant and distressing experience for expectant fathers who may benefit from an opportunity to express their childbirth-related fears in an environment where they feel validated and supported. Antenatal education is recommended to enhance fathers’ childbirth-related self-efficacy to reduce fear of childbirth. Conclusions: Fear of childbirth may negatively impact the lives of men and consequently their families. Further investigation into methods and models for identifying and supporting men at risk of or experiencing fear of childbirth is required to improve outcomes for this population of men.
Zoledronate Causes a Systemic Shift of Macrophage Polarization towards M1 In Vivo
Authors: Weber Manuel , Homm Andi , Müller Stefan , Frey Silke , Amann Kerstin , Ries Jutta , Geppert Carol , Preidl Raimund , Möst Tobias , Kämmerer Peer W. , Kesting Marco , Wehrhan Falk
Published: January 2021

Background: Immunomodulatory properties of bisphosphonates (BP) are suggested to contribute to the development of medication-associated osteonecrosis of the jaw (MRONJ). Furthermore, bisphosphonate-derived immune modulation might contribute to the anti-metastatic effect observed in breast cancer patients. Macrophages are potential candidates for the mediation of immunomodulatory effects of bisphosphonates. The study aimed to investigate the influence of bisphosphonates alone and in combination with surgical trauma on systemic macrophage polarization (M1 vs. M2) using an in vivo rat model. Methods: A total of 120 animals were divided into four groups. Groups 2 and 4 were treated with 8 × 40 μg/kg body weight of the BP Zoledronate i.p. (week 0–7). Groups 3 and 4 were exposed to surgical trauma (week 8, tooth extraction + tibia fracture), whereas in Group 1 neither medication nor surgical trauma was applied. After 8, 10, 12 and 16 weeks, skin, lung and spleen were immunohistochemically examined for macrophage polarization via expression analysis of CD68, CD163 and iNOS using a tissue microarray (TMA). Results: A significant shift of macrophage polarization towards M1 was observed in skin, spleen and lung tissue of animals, with and without surgical trauma, treated with BP when compared to those without BP application. Surgical trauma did not cause a significant increase towards M1 polarization. Conclusions: BP application leads to a systemic pro-inflammatory situation in vivo, independent of surgical trauma, as evidenced by the shift in macrophage polarization towards M1 in various somatic tissues. This provides a possible explanation for the clinically observed anti-tumor effect of bisphosphonates and might also contribute to pathogenesis of MRONJ.
The Expression of the Cancer-Associated lncRNA Snhg15 Is Modulated by EphrinA5-Induced Signaling
Authors: Pensold Daniel , Gehrmann Julia , Pitschelatow Georg , Walberg Asa , Braunsteffer Kai , Reichard Julia , Ravaei Amin , Linde Jenice , Lampert Angelika , Costa Ivan G. , Zimmer-Bensch Geraldine
Published: January 2021

The Eph receptor tyrosine kinases and their respective ephrin-ligands are an important family of membrane receptors, being involved in developmental processes such as proliferation, migration, and in the formation of brain cancer such as glioma. Intracellular signaling pathways, which are activated by Eph receptor signaling, are well characterized. In contrast, it is unknown so far whether ephrins modulate the expression of lncRNAs, which would enable the transduction of environmental stimuli into our genome through a great gene regulatory spectrum. Applying a combination of functional in vitro assays, RNA sequencing, and qPCR analysis, we found that the proliferation and migration promoting stimulation of mouse cerebellar granule cells (CB) with ephrinA5 diminishes the expression of the cancer-related lncRNA Snhg15. In a human medulloblastoma cell line (DAOY) ephrinA5 stimulation similarly reduced SNHG15 expression. Computational analysis identified triple-helix-mediated DNA-binding sites of Snhg15 in promoters of genes found up-regulated upon ephrinA5 stimulation and known to be involved in tumorigenic processes. Our findings propose a crucial role of Snhg15 downstream of ephrinA5-induced signaling in regulating gene transcription in the nucleus. These findings could be potentially relevant for the regulation of tumorigenic processes in the context of glioma.
Association of Sleep Architecture and Physiology with Depressive Disorder and Antidepressants Treatment
Authors: Hutka Peter , Krivosova Michaela , Muchova Zuzana , Tonhajzerova Ingrid , Hamrakova Andrea , Mlyncekova Zuzana , Mokry Juraj , Ondrejka Igor
Published: January 2021

Sleep problems are frequently associated with the principal diagnostic criteria for many mental disorders. Alterations in the sleep of depressive patients are of high clinical significance because continuous sleep problems raise the chance of relapse, recurrence, or suicide, as well as the need for augmenting medications. Most antidepressants have been proven to influence the sleep architecture. While some classes of antidepressants improve sleep, others may cause sleep impairment. The successful treatment of depressive disorder also requires an understanding of the effects of antidepressants on sleep. This article briefly reviews the physiology of sleep and the typical alterations in the sleep architecture in depressive patients and updates the different effects of the majority of antidepressants including novel drugs in clinical practice on sleep. The summary of the updated scientific findings of the relationship between depression and sleep disturbances could be clinically beneficial in choosing the best medication for depressive patients with concurrent sleep disorders.
The Puzzling Problem of Cardiolipin Membrane-Cytochrome c Interactions: A Combined Infrared and Fluorescence Study
Authors: Ripanti Francesca , Di Venere Almerinda , Cestelli Guidi Mariangela , Romani Martina , Filabozzi Alessandra , Carbonaro Marina , Piro Maria Cristina , Sinibaldi Federica , Nucara Alessandro , Mei Giampiero
Published: January 2021

The interaction of cytochrome c (cyt c) with natural and synthetic membranes is known to be a complex phenomenon, involving both protein and lipid conformational changes. In this paper, we combined infrared and fluorescence spectroscopy to study the structural transformation occurring to the lipid network of cardiolipin-containing large unilamellar vesicles (LUVs). The data, collected at increasing protein/lipid ratio, demonstrate the existence of a multi-phase process, which is characterized by: (i) the interaction of cyt c with the lipid polar heads; (ii) the lipid anchorage of the protein on the membrane surface; and (iii) a long-distance order/disorder transition of the cardiolipin acyl chains. Such effects have been quantitatively interpreted introducing specific order parameters and discussed in the frame of the models on cyt c activity reported in literature.
KV1.5–KVβ1.3 Recycling Is PKC-Dependent
Authors: Macias Alvaro , de la Cruz Alicia , Peraza Diego A. , Benito-Bueno Angela de , Gonzalez Teresa , Valenzuela Carmen
Published: January 2021

KV1.5 channel function is modified by different regulatory subunits. KVβ1.3 subunits assemble with KV1.5 channels and induce a fast and incomplete inactivation. Inhibition of PKC abolishes the KVβ1.3-induced fast inactivation, decreases the amplitude of the current KV1.5–KVβ1.3 and modifies their pharmacology likely due to changes in the traffic of KV1.5–KVβ1.3 channels in a PKC-dependent manner. In order to analyze this hypothesis, HEK293 cells were transfected with KV1.5–KVβ1.3 channels, and currents were recorded by whole-cell configuration of the patch-clamp technique. The presence of KV1.5 in the membrane was analyzed by biotinylation techniques, live cell imaging and confocal microscopy approaches. PKC inhibition resulted in a decrease of 33 ± 7% of channels in the cell surface due to reduced recycling to the plasma membrane, as was confirmed by confocal microscopy. Live cell imaging indicated that PKC inhibition almost abolished the recycling of the KV1.5–KVβ1.3 channels, generating an accumulation of channels into the cytoplasm. All these results suggest that the trafficking regulation of KV1.5–KVβ1.3 channels is dependent on phosphorylation by PKC and, therefore, they could represent a clinically relevant issue, mainly in those diseases that exhibit modifications in PKC activity.
Expression of Thomsen–Friedenreich Antigen in Colorectal Cancer and Association with Microsatellite Instability
Authors: Leão Beatriz , Wen Xiaogang , Duarte Henrique O. , Gullo Irene , Gonçalves Gilza , Pontes Patrícia , Castelli Claudia , Diniz Francisca , Mereiter Stefan , Gomes Joana , Carneiro Fátima , Reis Celso A.
Published: January 2021

Microsatellite instability (MSI) is a molecular phenotype due to a deficient DNA mismatch repair (dMMR). In colorectal cancer (CRC), dMMR/MSI is associated with several clinical and histopathological features, influences prognosis, and is a predictive factor of response to therapy. In daily practice, dMMR/MSI profiles are identified by immunohistochemistry and/or multiplex PCR. The Thomsen–Friedenreich (TF) antigen was previously found to be a potential single marker to identify MSI-high gastric cancers. Therefore, in this study, we aimed to disclose a possible association between TF expression and MSI status in CRC. Furthermore, we evaluated the relationship between TF expression and other clinicopathological features, including patient survival. We evaluated the expression of the TF antigen in a cohort of 25 MSI-high and 71 microsatellite stable (MSS) CRCs. No association was observed between the expression of the TF antigen and MSI-high status in CRC. The survival analysis revealed that patients with MSI-high CRC showed improved survival when the TF antigen was expressed. This finding holds promise as it indicates the potential use of the TF antigen as a biomarker of better prognosis in MSI-high CRCs that should be validated in an independent and larger CRC cohort.
HDAC6-Selective Inhibitor Overcomes Bortezomib Resistance in Multiple Myeloma
Authors: Lee Sang Wu , Yeon Soo-Keun , Kim Go Woon , Lee Dong Hoon , Jeon Yu Hyun , Yoo Jung , Kim So Yeon , Kwon So Hee
Published: January 2021

Although multiple myeloma (MM) patients benefit from standard bortezomib (BTZ) chemotherapy, they develop drug resistance, resulting in relapse. We investigated whether histone deacetylase 6 (HDAC6) inhibitor A452 overcomes bortezomib resistance in MM. We show that HDAC6-selective inhibitor A452 significantly decreases the activation of BTZ-resistant markers, such as extracellular signal-regulated kinases (ERK) and nuclear factor kappa B (NF-κB), in acquired BTZ-resistant MM cells. Combination treatment of A452 and BTZ or carfilzomib (CFZ) synergistically reduces BTZ-resistant markers. Additionally, A452 synergizes with BTZ or CFZ to inhibit the activation of NF-κB and signal transducer and activator of transcription 3 (STAT3), resulting in decreased expressions of low-molecular-mass polypeptide 2 (LMP2) and LMP7. Furthermore, combining A452 with BTZ or CFZ leads to synergistic cancer cell growth inhibition, viability decreases, and apoptosis induction in the BTZ-resistant MM cells. Overall, the synergistic effect of A452 with CFZ is more potent than that of A452 with BTZ in BTZ-resistant U266 cells. Thus, our findings reveal the HDAC6-selective inhibitor as a promising therapy for BTZ-chemoresistant MM.
An Adult Mouse Model of Dilated Cardiomyopathy Caused by Inducible Cardiac-Specific Bis Deletion
Authors: Yun Hye Hyeon , Jung Soon Young , Park Bong Woo , Ko Ji Seung , Yoo Kyunghyun , Yeo Jiyoung , Kim Hong Lim , Park Hun Jun , Youn Ho Joong , Lee Jeong Hwa
Published: January 2021

BCL-2 interacting cell death suppressor (BIS) is a multifunctional protein that has been implicated in cancer and myopathy. Various mutations of the BIS gene have been identified as causative of cardiac dysfunction in some dilated cardiomyopathy (DCM) patients. This was recently verified in cardiac-specific knock-out (KO) mice. In this study, we developed tamoxifen-inducible cardiomyocyte-specific BIS-KO (Bis-iCKO) mice to assess the role of BIS in the adult heart using the Cre-loxP strategy. The disruption of the Bis gene led to impaired ventricular function and subsequent heart failure due to DCM, characterized by reduced left ventricular contractility and dilatation that were observed using serial echocardiography and histology. The development of DCM was confirmed by alterations in Z-disk integrity and increased expression of several mRNAs associated with heart failure and remodeling. Furthermore, aggregation of desmin was correlated with loss of small heat shock protein in the Bis-iCKO mice, indicating that BIS plays an essential role in the quality control of cardiac proteins, as has been suggested in constitutive cardiac-specific KO mice. Our cardiac-specific BIS-KO mice may be a useful model for developing therapeutic interventions for DCM, especially late-onset DCM, based on the distinct phenotypes and rapid progressions.
BBS Proteins Affect Ciliogenesis and Are Essential for Hedgehog Signaling, but Not for Formation of iPSC-Derived RPE-65 Expressing RPE-Like Cells
Authors: Hey Caroline Amalie Brunbjerg , Larsen Lasse Jonsgaard , Tümer Zeynep , Brøndum-Nielsen Karen , Grønskov Karen , Hjortshøj Tina Duelund , Møller Lisbeth Birk
Published: January 2021

Bardet-Biedl syndrome (BBS) is a ciliopathy characterized by retinal dystrophy, renal cysts, obesity and polydactyly. BBS genes have been implicated in ciliogenesis, hedgehog signaling and retinal pigment epithelium maturation. BBS1 and BBS5 are members of the BBSome, implicated in cilia transport of proteins, and BBS10 is a member of the chaperonin-complex, mediating BBSome assembly. In this study, involvement of BBS1, BBS5 and BBS10 in ciliogenesis and hedgehog signaling were investigated in BBS-defective patient fibroblasts as well as in RPE-hTERT cells following siRNA-mediated knockdown of the BBS genes. Furthermore, the ability of BBS1-defective induced pluripotent stem-cells (iPSCs) to differentiate into RPE cells was assessed. We report that cells lacking functional BBS5 or BBS10 have a reduced number of primary cilia, whereas cells lacking functional BBS1 display shorter primary cilia compared to wild-type cells. Hedgehog signaling was substantially impaired and Smoothened, a component of hedgehog signaling, was trapped inside the cilia of the BBS-defective cells, even in the absence of Smoothened agonist. Preliminary results demonstrated the ability of BBS1-defective iPSC to differentiate into RPE-65 expressing RPE-like cells. The BBS1−/−-defective RPE-like cells were less pigmented, compared to RPE-like cells differentiated from control iPSCs, indicating an impact of BBS1 on RPE maturation.
Imprinted Genes and Multiple Sclerosis: What Do We Know?
Authors: Baulina Natalia , Kiselev Ivan , Favorova Olga
Published: January 2021

Multiple sclerosis (MS) is a chronic autoimmune neurodegenerative disease of the central nervous system that arises from interplay between non-genetic and genetic risk factors. The epigenetics functions as a link between these factors, affecting gene expression in response to external influence, and therefore should be extensively studied to improve the knowledge of MS molecular mechanisms. Among others, the epigenetic mechanisms underlie the establishment of parent-of-origin effects that appear as phenotypic differences depending on whether the allele was inherited from the mother or father. The most well described manifestation of parent-of-origin effects is genomic imprinting that causes monoallelic gene expression. It becomes more obvious that disturbances in imprinted genes at the least affecting their expression do occur in MS and may be involved in its pathogenesis. In this review we will focus on the potential role of imprinted genes in MS pathogenesis.
Folinate Supplementation Ameliorates Methotrexate Induced Mitochondrial Formate Depletion In Vitro and In Vivo
Authors: Sou Nga-Lai , Huang Yu-Hsuan , Chen Der-Yuan , Chen Yi-Ming , Tang Feng-Yao , Ko Hsin-An , Fan Yi-Hsuan , Lin Yi-Ying , Wang Yi-Cheng , Chih Hui-Ming , Shane Barry , Huang Wen-Nan , Chiang En-Pei Isabel
Published: January 2021

(1) Background: Antifolate methotrexate (MTX) is the most common disease-modifying antirheumatic drug (DMARD) for treating human rheumatoid arthritis (RA). The mitochondrial-produced formate is essential for folate-mediated one carbon (1C) metabolism. The impacts of MTX on formate homeostasis in unknown, and rigorously controlled kinetic studies can greatly help in this regard. (2) Methods: Combining animal model (8-week old female C57BL/6JNarl mice, n = 18), cell models, stable isotopic tracer studies with gas chromatography/mass spectrometry (GC/MS) platforms, we systematically investigated how MTX interferes with the partitioning of mitochondrial and cytosolic formate metabolism. (3) Results: MTX significantly reduced de novo deoxythymidylate (dTMP) and methionine biosyntheses from mitochondrial-derived formate in cells, mouse liver, and bone marrow, supporting our postulation that MTX depletes mitochondrial 1C supply. Furthermore, MTX inhibited formate generation from mitochondria glycine cleavage system (GCS) both in vitro and in vivo. Folinate selectively rescued 1C metabolic pathways in a tissue-, cellular compartment-, and pathway-specific manner: folinate effectively reversed the inhibition of mitochondrial formate-dependent 1C metabolism in mouse bone marrow (dTMP, methionine, and GCS) and cells (dTMP and GCS) but not methionine synthesis in liver/liver-derived cells. Folinate failed to fully recover hepatic mitochondrial-formate utilization for methionine synthesis, suggesting that the efficacy of clinical folinate rescue in MTX therapy on hepatic methionine metabolism is poor. (4) Conclusion: Conducting studies in mouse and cell models, we demonstrate novel findings that MTX specifically depletes mitochondrial 1C supply that can be ameliorated by folinate supplementation except for hepatic transmethylation. These results imply that clinical use of low-dose MTX may particularly impede 1C metabolism via depletion of mitochondrial formate. The MTX induced systematic and tissue-specific formate depletion needs to be addressed more carefully, and the efficacy of folinate with respect to protecting against such depletion deserves to be evaluated in medical practice.
A Pipeline towards the Biochemical Characterization of the Arabidopsis GT14 Family
Authors: Xuan Lingling , Zhang Jie , Lu Weitai , Gluza Pawel , Ebert Berit , Kotake Toshihisa , Lu Mengzhu , Zhang Yuan , Clausen Mads H. , Johnson Kim L. , Doblin Monika S. , Heazlewood Joshua L. , Bacic Antony , Song Lili , Zeng Wei
Published: January 2021

Glycosyltransferases (GTs) catalyze the synthesis of glycosidic linkages and are essential in the biosynthesis of glycans, glycoconjugates (glycolipids and glycoproteins), and glycosides. Plant genomes generally encode many more GTs than animal genomes due to the synthesis of a cell wall and a wide variety of glycosylated secondary metabolites. The Arabidopsis thaliana genome is predicted to encode over 573 GTs that are currently classified into 42 diverse families. The biochemical functions of most of these GTs are still unknown. In this study, we updated the JBEI Arabidopsis GT clone collection by cloning an additional 105 GT cDNAs, 508 in total (89%), into Gateway-compatible vectors for downstream characterization. We further established a functional analysis pipeline using transient expression in tobacco (Nicotiana benthamiana) followed by enzymatic assays, fractionation of enzymatic products by reversed-phase HPLC (RP-HPLC) and characterization by mass spectrometry (MS). Using the GT14 family as an exemplar, we outline a strategy for identifying effective substrates of GT enzymes. By addition of UDP-GlcA as donor and the synthetic acceptors galactose-nitrobenzodiazole (Gal-NBD), β-1,6-galactotetraose (β-1,6-Gal4) and β-1,3-galactopentose (β-1,3-Gal5) to microsomes expressing individual GT14 enzymes, we verified the β-glucuronosyltransferase (GlcAT) activity of three members of this family (AtGlcAT14A, B, and E). In addition, a new family member (AT4G27480, 248) was shown to possess significantly higher activity than other GT14 enzymes. Our data indicate a likely role in arabinogalactan-protein (AGP) biosynthesis for these GT14 members. Together, the updated Arabidopsis GT clone collection and the biochemical analysis pipeline present an efficient means to identify and characterize novel GT catalytic activities.
Cancer-Targeted Controlled Delivery of Chemotherapeutic Anthracycline Derivatives Using Apoferritin Nanocage Carriers
Authors: Kurzątkowska Katarzyna , Pazos Manuel A. , Herschkowitz Jason I. , Hepel Maria
Published: January 2021

The interactions of chemotherapeutic drugs with nanocage protein apoferritin (APO) are the key features in the effective encapsulation and release of highly toxic drugs in APO-based controlled drug delivery systems. The encapsulation enables mitigating the drugs’ side effects, collateral damage to healthy cells, and adverse immune reactions. Herein, the interactions of anthracycline drugs with APO were studied to assess the effect of drug lipophilicity on their encapsulation excess n and in vitro activity. Anthracycline drugs, including doxorubicin (DOX), epirubicin (EPI), daunorubicin (DAU), and idarubicin (IDA), with lipophilicity P from 0.8 to 15, were investigated. We have found that in addition to hydrogen-bonded supramolecular ensemble formation with n = 24, there are two other competing contributions that enable increasing n under strong polar interactions (APO(DOX)) or under strong hydrophobic interactions (APO(IDA) of the highest efficacy). The encapsulation/release processes were investigated using UV-Vis, fluorescence, circular dichroism, and FTIR spectroscopies. The in vitro cytotoxicity/growth inhibition tests and flow cytometry corroborate high apoptotic activity of APO(drugs) against targeted MDA-MB-231 adenocarcinoma and HeLa cells, and low activity against healthy MCF10A cells, demonstrating targeting ability of nanodrugs. A model for molecular interactions between anthracyclines and APO nanocarriers was developed, and the relationships derived compared with experimental results.
Brain-Derived Extracellular Vesicles in Health and Disease: A Methodological Perspective
Authors: Brenna Santra , Krisp Christoph , Altmeppen Hermann Clemens , Magnus Tim , Puig Berta
Published: January 2021

Extracellular vesicles (EVs) are double membrane structures released by presumably all cell types that transport and deliver lipids, proteins, and genetic material to near or distant recipient cells, thereby affecting their phenotype. The basic knowledge of their functions in healthy and diseased brain is still murky and many questions about their biology are unsolved. In neurological diseases, EVs are regarded as attractive biomarkers and as therapeutic tools due to their ability to cross the blood–brain barrier (BBB). EVs have been successfully isolated from conditioned media of primary brain cells and cerebrospinal fluid (CSF), but protocols allowing for the direct study of pathophysiological events mediated or influenced by EVs isolated from brain have only recently been published. This review aims to give a brief overview of the current knowledge of EVs’ functions in the central nervous system (CNS) and the current protocols to isolate brain-derived EVs (BDEVs) used in different publications. By comparing the proteomic analysis of some of these publications, we also assess the influence of the isolation method on the protein content of BDEVs.
Artemisia annua L. Polyphenol-Induced Cell Death Is ROS-Independently Enhanced by Inhibition of JNK in HCT116 Colorectal Cancer Cells
Authors: Jung Eun Joo , Paramanantham Anjugam , Kim Hye Jung , Shin Sung Chul , Kim Gon Sup , Jung Jin-Myung , Ryu Chung Ho , Hong Soon Chan , Chung Ky Hyun , Kim Choong Won , Lee Won Sup
Published: January 2021

c-Jun N-terminal kinase (JNK) is activated by chemotherapeutic reagents including natural plant polyphenols, and cell fate is determined by activated phospho-JNK as survival or death depending on stimuli and cell types. The purpose of this study was to elucidate the role of JNK on the anticancer effects of the Korean plant Artemisia annua L. (pKAL) polyphenols in p53 wild-type HCT116 human colorectal cancer cells. Cell morphology, protein expression levels, apoptosis/necrosis, reactive oxygen species (ROS), acidic vesicles, and granularity/DNA content were analyzed by phase-contrast microscopy; Western blot; and flow cytometry of annexin V/propidium iodide (PI)-, dichlorofluorescein (DCF)-, acridine orange (AO)-, and side scatter pulse height (SSC-H)/DNA content (PI)-stained cells. The results showed that pKAL induced morphological changes and necrosis or late apoptosis, which were associated with loss of plasma membrane/Golgi integrity, increased acidic vesicles and intracellular granularity, and decreased DNA content through downregulation of protein kinase B (Akt)/β-catenin/cyclophilin A/Golgi matrix protein 130 (GM130) and upregulation of phosphorylation of H2AX at Ser-139 (γ-H2AX)/p53/p21/Bak cleavage/phospho-JNK/p62/microtubule-associated protein 1 light chain 3B (LC3B)-I. Moreover, JNK inhibition by SP600125 enhanced ROS-independently pKAL-induced cell death through downregulation of p62 and upregulation of p53/p21/Bak cleavage despite a reduced state of DNA damage marker γ-H2AX. These findings indicate that phospho-JNK activated by pKAL inhibits p53-dependent cell death signaling and enhances DNA damage signaling, but cell fate is determined by phospho-JNK as survival rather than death in p53 wild-type HCT116 cells.
Moonlighting Proteins Shine New Light on Molecular Signaling Niches
Authors: Turek Ilona , Irving Helen
Published: January 2021

Plants as sessile organisms face daily environmental challenges and have developed highly nuanced signaling systems to enable suitable growth, development, defense, or stalling responses. Moonlighting proteins have multiple tasks and contribute to cellular signaling cascades where they produce additional variables adding to the complexity or fuzziness of biological systems. Here we examine roles of moonlighting kinases that also generate 3′,5′-cyclic guanosine monophosphate (cGMP) in plants. These proteins include receptor like kinases and lipid kinases. Their guanylate cyclase activity potentiates the development of localized cGMP-enriched nanodomains or niches surrounding the kinase and its interactome. These nanodomains contribute to allosteric regulation of kinase and other molecules in the immediate complex directly or indirectly modulating signal cascades. Effects include downregulation of kinase activity, modulation of other members of the protein complexes such as cyclic nucleotide gated channels and potential triggering of cGMP-dependent degradation cascades terminating signaling. The additional layers of information provided by the moonlighting kinases are discussed in terms of how they may be used to provide a layer of fuzziness to effectively modulate cellular signaling cascades.
Acknowledgment to Reviewers of Instruments in 2020
Authors:   Instruments Editorial Office
Published: January 2021

Peer review is the driving force of journal development, and reviewers are gatekeepers who ensure that Instruments maintains its standards for the high quality of its published papers [...]