bluuebunny/crossref_metadata_2025 huggingface dataset:
Understanding Knowledge and Behaviors Related to CoViD–19 Epidemic in Italian Undergraduate Students: The EPICO Study
Authors: Gallè Francesca , Sabella Elita Anna , Da Molin Giovanna , De Giglio Osvalda , Caggiano Giuseppina , Di Onofrio Valeria , Ferracuti Stefano , Montagna Maria Teresa , Liguori Giorgio , Orsi Giovanni Battista , Napoli Christian
Published: May 2020

Background: On February 2020, the novel coronavirus (2019−nCoV) epidemic began in Italy. In order to contain the spread of the virus, the Italian government adopted emergency measures nationwide, including closure of schools and universities, workplaces and subsequently lockdown. This survey was carried out among Italian undergraduates to explore their level of knowledge about the epidemic and the behaviors they adopted during the lockdown. Methods: An electronic questionnaire was administered to the students attending three Italian universities. Results: A good level of knowledge about the epidemic and its control was registered in the sample, mainly among students attending life sciences degree courses. The majority of the students did not modify their diet and smoking habits, while a great part of the sample reported a decrease in physical activity (PA). Conclusions: Students from life sciences courses showed a higher awareness regarding the infection and the control measures. The lockdown caused an important reduction of PA. Preventive interventions should transform the restrictive measures also as an opportunity to improve lifestyle.
Distribution of the SARS-CoV-2 Pandemic and Its Monthly Forecast Based on Seasonal Climate Patterns
Authors: Scafetta Nicola
Published: May 2020

This paper investigates whether the Severe Acute Respiratory Syndrome CoronaVirus 2 (SARS-CoV-2) pandemic could have been favored by specific weather conditions and other factors. It is found that the 2020 winter weather in the region of Wuhan (Hubei, Central China)—where the virus first broke out in December and spread widely from January to February 2020—was strikingly similar to that of the Northern Italian provinces of Milan, Brescia and Bergamo, where the pandemic broke out from February to March. The statistical analysis was extended to cover the United States of America, which overtook Italy and China as the country with the highest number of confirmed COronaVIrus Disease 19 (COVID-19) cases, and then to the entire world. The found correlation patterns suggest that the COVID-19 lethality significantly worsens (4 times on average) under weather temperatures between 4 °C and 12 °C and relative humidity between 60% and 80%. Possible co-factors such as median population age and air pollution were also investigated suggesting an important influence of the former but not of the latter, at least, on a synoptic scale. Based on these results, specific isotherm world maps were generated to locate, month by month, the world regions that share similar temperature ranges. From February to March, the 4–12 °C isotherm zone extended mostly from Central China toward Iran, Turkey, West-Mediterranean Europe (Italy, Spain and France) up to the United State of America, optimally coinciding with the geographic regions most affected by the pandemic from February to March. It is predicted that in the spring, as the weather gets warm, the pandemic will likely worsen in northern regions (United Kingdom, Germany, East Europe, Russia and North America) while the situation will likely improve in the southern regions (Italy and Spain). However, in autumn, the pandemic could come back and affect the same regions again. The Tropical Zone and the entire Southern Hemisphere, but in restricted colder southern regions, could avoid a strong pandemic because of the sufficiently warm weather during the entire year and because of the lower median age of their population. Google-Earth-Pro interactive-maps covering the entire world are provided as supplementary files.
Emerging Adults and COVID-19: The Role of Individualism-Collectivism on Perceived Risks and Psychological Maladjustment
Authors: Germani Alessandro , Buratta Livia , Delvecchio Elisa , Mazzeschi Claudia
Published: May 2020

The outbreak of the coronavirus disease 2019 (COVID-19) has dramatically changed our habits and routines. Uncertainty, insecurity, instability for the present and future, and reduced autonomy and self-directedness, are common feelings at the time of COVID-19. These aspects are very important during emerging adulthood. In spite of the fact that medical reports suggest that youth are less prone to experience COVID-19 infections, emerging adults might be at higher risk for their psychological adjustment. Emerging adults showed higher concerns about their role as a possible asymptomatic carrier than being positive with COVID-19 themselves. Both worries and concerns about COVID-19 and psychological maladjustment may be related to cultural factors. Individualism, collectivism, equality, and hierarchy seem to be meaningful perspectives to take into account. A total of 1183 Italian emerging adults were asked to fill out an online survey during the second week of the national lockdown in Italy. Results showed they reported an accurate perceived knowledge about COVID-19. At the same time, they showed higher worries and concerns about COVID-19 for their relatives, followed by more general/social worries. The lowest score included worries about COVID-19 related to themselves. State anxiety and stress levels were above the normal cutoff, confirming the challenges that emerging adults are facing during the pandemic. On one hand, emerging adults’ collectivistic orientation was related to higher perceived risks of infection; on the other hand, it predicted lower psychological maladjustment, controlling for socio-demographic variables. The study suggests that to fight the COVID-19 pandemic and decrease levels of psychological maladjustment in emerging adulthood, individuals’ cultural orientation such as the wish of sharing common goals with others, interdependence, and sociability, have to be emphasized and promoted as protective factors.
Robotic Technology in Pediatric Neurorehabilitation. A Pilot Study of Human Factors in an Italian Pediatric Hospital
Authors: Gilardi Francesco , De Falco Federica , Casasanta Daniela , Andellini Martina , Gazzellini Simone , Petrarca Maurizio , Morocutti Andreina , Lettori Donatella , Ritrovato Matteo , Castelli Enrico , Raponi Massimiliano , Magnavita Nicola , Zaffina Salvatore
Published: May 2020

The introduction of robotic neurorehabilitation among the most recent technologies in pediatrics represents a new opportunity to treat pediatric patients. This study aims at evaluating the response of physiotherapists, patients and their parents to this new technology. The study considered the outcomes of technological innovation in physiotherapists (perception of the workload, satisfaction), as well as that in patients and their parents (quality of life, expectations, satisfaction) by comparing the answers to subjective questionnaires of those who made use of the new technology with those who used the traditional therapy. A total of 12 workers, 46 patients and 47 parents were enrolled in the study. Significant differences were recorded in the total workload score of physiotherapists who use the robotic technology compared with the traditional therapy (p < 0.001). Patients reported a higher quality of life and satisfaction after the use of the robotic neurorehabilitation therapy. The parents of patients undergoing the robotic therapy have moderately higher expectations and satisfaction than those undergoing the traditional therapy. In this pilot study, the robotic neurorehabilitation technique involved a significant increase in the patients’ and parents’ expectations. As it frequently happens in the introduction of new technologies, physiotherapists perceived a greater workload. Further studies are needed to verify the results achieved.
A Brief Incubation of Cumulus-Enclosed Mouse Eggs in a Calcium-Free Medium Containing a High Concentration of Calcium-Chelator Markedly Improves Preimplantation Development
Authors: Merico Valeria , Garagna Silvia , Zuccotti Maurizio
Published: May 2020

The presence of cumulus cells (CCs) surrounding ovulated eggs is beneficial to in vitro fertilization and preimplantation development outcomes in several mammalian species. In the mouse, this contribution has a negligible effect on the fertilization rate; however, it is not yet clear whether it has positive effects on preimplantation development. Here, we compared the rates of in vitro fertilization and preimplantation development of ovulated B6C3F1 CC-enclosed vs. CC-free eggs, the latter obtained either after a 5 min treatment in M2 medium containing hyaluronidase or after 5–25 min in M2 medium supplemented with 34.2 mM EDTA (M2-EDTA). We found that, although the maintenance of CCs around ovulated eggs does not increment their developmental rate to blastocyst, the quality of the latter is significantly enhanced. Most importantly, for the first time, we describe a further quantitative and qualitative improvement, on preimplantation development, when CC-enclosed eggs are isolated from the oviducts in M2-EDTA and left in this medium for a total of 5 min prior to sperm insemination. Altogether, our results establish an important advancement in mouse IVF procedures that would be now interesting to test on other mammalian species.
Methodological Issues in the Study of the Development of Pain Responsivity in Preterm Neonates: A Systematic Review
Authors: Menin Damiano , Dondi Marco
Published: May 2020

The study of the development of neonatal pain responses is of key importance, both for research and for clinical reasons, with particular regard to the population of preterm neonates, given the amount of painful procedures they are exposed to on a daily basis. The aim of this work was to systematize our knowledge about the development of pain responses in prematurely born neonates by focusing on some key methodological issues. Studies on the impact of age variables, namely gestational age (GA), postmenstrual age (PMA) and chronological age (CH), on pain responsivity in premature neonates were identified using Medline and Scopus. Studies (N = 42) were categorized based on terminological and methodological approaches towards age variables, and according to output variables considered (facial, nonfacial behavioral, physiological). Distinct multidimensional developmental patterns were found for each age-sampling strategy. Overall, each of the three age variables seems to affect pain responsivity, possibly differently across age windows. Targeted as well as integrated approaches, together with a renewed attention for methodological consistency, are needed to further our knowledge on this topic.
Are There Effective Interventions to Increase Physical Activity in Children and Young People? An Umbrella Review
Authors: Mannocci Alice , D’Egidio Valeria , Backhaus Insa , Federici Antonio , Sinopoli Alessandra , Ramirez Varela Andrea , Villari Paolo , La Torre Giuseppe
Published: May 2020

Background: Obesity and physical inactivity among children and young people are public health concerns. While numerous interventions to promote physical activity are available, little is known about the most effective ones. This study aimed to summarize the existing evidence on interventions that aim to increase physical activity. Methods: A systematic review of reviews was conducted. Systematic reviews and meta-analyses published from January 2010 until November 2017 were identified through PubMed, Scopus and the Cochrane Library. Two reviewers independently assessed titles and abstracts, performed data extraction and quality assessment. Outcomes as level of physical activity and body mass index were collected in order to assess the efficacy of interventions. Results: A total 30 studies examining physical activity interventions met the inclusion criteria, 15 systematic reviews and 15 meta-analyses. Most studies (N = 20) were implemented in the school setting, three were developed in preschool and childcare settings, two in the family context, five in the community setting and one miscellaneous context. Results showed that eight meta-analyses obtained a small increase in physical activity level, out of which five were conducted in the school, two in the family and one in the community setting. Most promising programs had the following characteristics: included physical activity in the school curriculum, were long-term interventions, involved teachers and had the support of families. Conclusion: The majority of interventions to promote physical activity in children and young people were implemented in the school setting and were multicomponent. Further research is needed to investigate nonschool programs.
Ranking of Assets with Respect to Their Exposure to the Landslide Hazard: A GIS Proposal
Authors: Di Felice Paolino
Published: May 2020

The need to protect critical infrastructures (for short called assets within this paper) arises because of the hazards they are exposed to. In this article, the hazard is represented by the landslides. The first part of the paper proposes a scientifically robust method for the identification of the top-N assets that can be modeled as “points” (mainly buildings). The developed method takes into account the slope of the terrain, the runout distance of the landslide and its trajectory. The latter is roughly estimated through the notion of linear regression line. The method is applied to a real case to carry out a preliminary validation of it. In the second part of the paper, it is formalized the problem of computing the ranking of assets that can be modeled as “lines” (e.g., highways, power lines, pipelines, railway lines, and so on, that cross a given territory). The problem is solved in three steps: (a) Segmentation (it “cuts” each route in segments), (b) Sampling (it extracts points from each segment), and (c) Calculation (it associates an exposure value to each extracted point and, then, computes the exposure of the various segments composing the routes). The computation of the exposure for the points is carried out by applying the method of the first part of the paper. Both rankings can be used by the local administrators as a conceptual tool for narrowing down a global problem to smaller, higher exposure, geographic areas where the management of the hazard is crucial.
Optimal Lowest Astronomical Tide Estimation Using Maximum Likelihood Estimator with Multiple Ocean Models Hybridization
Authors: El-Diasty Mohammed
Published: May 2020

Developing an accurate Lowest Astronomical Tide (LAT) in a continuous form is essential for many maritime applications as it can be employed to develop an accurate continuous vertical control datum for hydrographic surveys applications and to produce accurate dynamic electronic navigation charts for safe maritime navigation by mariners. The LAT can be developed in a continuous (surface) using an estimated LAT surface model from the hydrodynamic ocean model along with coastal discrete LAT point values derived from tide gauges data sets to provide the corrected LAT surface model. In this paper, an accurate LAT surface model was developed for the Red Sea case study using a Maximum Likelihood Estimator (MLE) with multiple hydrodynamic ocean models hybridization, namely, WebTide, FES2014, DTU10, and EOT11a models. It was found that the developed optimal hybrid LAT model using MLE with multiple hydrodynamic ocean models hybridization ranges from 0.1 m to 1.63 m, associated with about 2.4 cm of uncertainty at a 95% confidence level in the Red Sea case study area. To validate the accuracy of the developed model, the comparison was made between the optimal hybrid LAT model developed from multiple hydrodynamic ocean models hybridization using the MLE method with the individual LAT models estimated from individual WebTide, FES2014, DTU10, or EOT11a ocean models based on the associated uncertainties estimated at a 95% confidence level. It was found that the optimal hybrid LAT model accuracy is superior to the individual LAT models estimated from individual ocean models with an improvement of about 50% in average, based on the estimated uncertainties. The importance of developing optimal LAT surface model using the MLE method with multiple hydrodynamic ocean models hybridization in this paper with few centimeters level of uncertainty can lead to accurate continuous vertical datum estimation that is essential for many maritime applications.
Suppression of PKCδ/NF-κB Signaling and Apoptosis Induction through Extrinsic/Intrinsic Pathways Are Associated with Magnolol-Inhibited Tumor Progression in Colorectal Cancer In Vitro and In Vivo
Authors: Su Chun-Min , Weng Yueh-Shan , Kuan Lin-Yen , Chen Jiann-Hwa , Hsu Fei-Ting
Published: May 2020

Magnolol is one of the hydroxylated biphenyl compounds from the root and stem bark of Magnolia officinalis, which shown to possess anti-colorectal cancer (CRC) effects. However, the regulatory mechanism of magnolol on apoptosis and NF-κB signaling in human CRC has not been elucidated. Thus, we investigated the inhibitory mechanism of magnolol on human and mouse CRC (HT-29 and CT-26) in vitro and in vivo. Results from reporter gene assay indicated that both magnolol and rottlerin (PKCδ inhibitor) reduced the endogenous NF-κB activity. In addition, indolactam V (PKCδ activator)-induced NF-κB signaling was significantly suppressed with both magnolol and rottlerin treatment. Results from Western blotting also indicated that phosphorylation of PKCδ and NF-κB -related proteins involved in tumor progression were effectively decreased by magnolol treatment. The invasion capacity of CRC cells was also attenuated by both magnolol and rottlerin. Furthermore, magnolol triggered Fas/Fas-L mediated extrinsic apoptosis and mitochondria mediated intrinsic apoptosis were validated by flow cytometry. Most importantly, tumor growth in both HT-29 and CT-26 bearing mice were suppressed by magnolol, but no pathologic change was detected in mice kidney, spleen, and liver. As confirmed by immunohistochemistry (IHC) staining from tumor tissue, PKCδ/NF-κB signaling and downstream proteins expression were decreased, while apoptotic proteins expression was increased in the magnolol treated group. According to these results, we suggest that the induction of apoptosis through extrinsic/intrinsic pathways and the blockage of PKCδ/NF-κB signaling are associated with the magnolol-inhibited progression of CRC.
Soil Microbiome Response to Contamination with Bisphenol A, Bisphenol F and Bisphenol S
Authors: Zaborowska Magdalena , Wyszkowska Jadwiga , Borowik Agata
Published: May 2020

The choice of the study objective was affected by numerous controversies and concerns around bisphenol F (BPF) and bisphenol S (BPS)—analogues of bisphenol A (BPA). The study focused on the determination and comparison of the scale of the BPA, BPF, and BPS impact on the soil microbiome and its enzymatic activity. The following parameters were determined in soil uncontaminated and contaminated with BPA, BPF, and BPS: the count of eleven groups of microorganisms, colony development (CD) index, microorganism ecophysiological diversity (EP) index, genetic diversity of bacteria and activity of dehydrogenases (Deh), urease (Ure), catalase (Cat), acid phosphatase (Pac), alkaline phosphatase (Pal), arylsulphatase (Aryl) and β-glucosidase (Glu). Bisphenols A, S and F significantly disrupted the soil homeostasis. BPF is regarded as the most toxic, followed by BPS and BPA. BPF and BPS reduced the abundance of Proteobacteria and Acidobacteria and increased that of Actinobacteria. Unique types of bacteria were identified as well as the characteristics of each bisphenol: Lysobacter, Steroidobacter, Variovorax, Mycoplana, for BPA, Caldilinea, Arthrobacter, Cellulosimicrobium and Promicromonospora for BPF and Dactylosporangium Geodermatophilus, Sphingopyxis for BPS. Considering the strength of a negative impact of bisphenols on the soil biochemical activity, they can be arranged as follows: BPS > BPF > BPA. Urease and arylsulphatase proved to be the most susceptible and dehydrogenases the least susceptible to bisphenols pressure, regardless of the study duration.
Dual Deep Sequencing Improves the Accuracy of Low-Frequency Somatic Mutation Detection in Cancer Gene Panel Testing
Authors: Ura Hiroki , Togi Sumihito , Niida Yo
Published: May 2020

Cancer gene panel testing requires accurate detection of somatic mosaic mutations, as the test sample consists of a mixture of cancer cells and normal cells; each minor clone in the tumor also has different somatic mutations. Several studies have shown that the different types of software used for variant calling for next generation sequencing (NGS) can detect low-frequency somatic mutations. However, the accuracy of these somatic variant callers is unknown. We performed cancer gene panel testing in duplicate experiments using three different high-fidelity DNA polymerases in pre-capture amplification steps and analyzed by three different variant callers, Strelka2, Mutect2, and LoFreq. We selected six somatic variants that were detected in both experiments with more than two polymerases and by at least one variant caller. Among them, five single nucleotide variants were verified by CEL nuclease-mediated heteroduplex incision with polyacrylamide gel electrophoresis and silver staining (CHIPS) and Sanger sequencing. In silico analysis indicated that the FBXW7 and MAP3K1 missense mutations cause damage at the protein level. Comparing three somatic variant callers, we found that Strelka2 detected more variants than Mutect2 and LoFreq. We conclude that dual sequencing with Strelka2 analysis is useful for detection of accurate somatic mutations in cancer gene panel testing.
Current Molecular Markers of Melanoma and Treatment Targets
Authors: Yang Kevin , Oak Allen S.W. , Slominski Radomir M. , Brożyna Anna A. , Slominski Andrzej T.
Published: May 2020

Melanoma is a deadly skin cancer that becomes especially difficult to treat after it metastasizes. Timely identification of melanoma is critical for effective therapy, but histopathologic diagnosis can frequently pose a significant challenge to this goal. Therefore, auxiliary diagnostic tools are imperative to facilitating prompt recognition of malignant lesions. Melanoma develops as result of a number of genetic mutations, with UV radiation often acting as a mutagenic risk factor. Novel methods of genetic testing have improved detection of these molecular alterations, which subsequently revealed important information for diagnosis and prognosis. Rapid detection of genetic alterations is also significant for choosing appropriate treatment and developing targeted therapies for melanoma. This review will delve into the understanding of various mutations and the implications they may pose for clinical decision making.
The Role of DNA Degradation in the Estimation of Post-Mortem Interval: A Systematic Review of the Current Literature
Authors: Tozzo Pamela , Scrivano Salvatore , Sanavio Matteo , Caenazzo Luciana
Published: May 2020

The determination of the post-mortal interval (PMI) is an extremely discussed topic in the literature and of deep forensic interest, for which various types of methods have been proposed. The aim of the manuscript is to provide a review of the studies on the post-mortem DNA degradation used for estimating PMI. This review has been performed following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses and the PRISMA Guidelines. Several analytical techniques have been proposed to analyse the post-mortem DNA degradation in order to use it to estimate the PMI. Studies focused mainly on animal models and on particular tissues. The results have been mixed: while on the one hand literature data in this field have confirmed that in the post-mortem several degradation processes involve nucleic acids, on the other hand some fundamental aspects are still little explored: the influence of ante and post-mortem factors on DNA degradation, the feasibility and applicability of a multiparametric mathematical model that takes into account DNA degradation and the definition of one or more target organs in order to standardize the results on human cases under standard conditions.
Exosomal MiRNA Transfer between Retinal Microglia and RPE
Authors: Morris Dorothea R. , Bounds Sarah E. , Liu Huanhuan , Ding Wei-Qun , Chen Yan , Liu Yin , Cai Jiyang
Published: May 2020

The retinal pigment epithelium (RPE), the outermost layer of the retina, provides essential support to both the neural retina and choroid. Additionally, the RPE is highly active in modulating functions of immune cells such as microglia, which migrate to the subretinal compartment during aging and age-related degeneration. Recently, studies have highlighted the important roles of microRNA (miRNA) in the coordination of general tissue maintenance as well as in chronic inflammatory conditions. In this study, we analyzed the miRNA profiles in extracellular vesicles (EVs) released by the RPE, and identified and validated miRNA species whose expression levels showed age-dependent changes in the EVs. Using co-culture of RPE and retinal microglia, we further demonstrated that miR-21 was transferred between the two types of cells, and the increased miR-21 in microglia influenced the expression of genes downstream of the p53 pathway. These findings suggest that exosome-mediated miRNA transfer is a signaling mechanism that contributes to the regulation of microglia function in the aging retina.
Complete Mitochondrial Genomes of Two Toxin-Accumulated Nassariids (Neogastropoda: Nassariidae: Nassarius) and Their Implication for Phylogeny
Authors: Yang Yi , Liu Hongyue , Qi Lu , Kong Lingfeng , Li Qi
Published: May 2020

The Indo-Pacific nassariids (genus Nassarius) possesses the highest diversity within the family Nassariidae. However, the previous shell or radula-based classification of Nassarius is quite confusing due to the homoplasy of certain morphological characteristics. The toxin accumulators Nassarius glans and Nassarius siquijorensis are widely distributed in the subtidal regions of the Indo-Pacific Ocean. In spite of their biological significance, the phylogenetic positions of N. glans and N. siquijorensis are still undetermined. In the present study, the complete mitochondrial genomes of N. glans and N. siquijorensis were sequenced. The present mitochondrial genomes were 15,296 and 15,337 bp in length, respectively, showing negative AT skews and positive GC skews as well as a bias of AT rich on the heavy strand. They contained 13 protein coding genes, 22 transfer RNA genes, two ribosomal RNA genes, and several noncoding regions, and their gene order was identical to most caenogastropods. Based on the nucleotide sequences combining 13 protein coding genes and two rRNA genes, a well-supported phylogeny of Nassarius was reconstructed, and several morphological synapomorphies were observed corresponding to the phylogenetic framework. In addition, the sister group relationship between N. variciferus and the remaining toxin-accumulated nassariids was determined, suggesting that the phylogeny might be related to their diet. The divergence time estimation analysis revealed a correlation between speciation events of nassariids and glacial cycles during the Pliocene-Pleistocene epoch.
The MKK-Dependent Phosphorylation of p38α Is Augmented by Arginine Methylation on Arg49/Arg149 during Erythroid Differentiation
Authors: Liu Mei-Yin , Hua Wei-Kai , Chen Chi-Ju , Lin Wey-Jinq
Published: May 2020

The activation of p38 mitogen-activated protein kinases (MAPKs) through a phosphorylation cascade is the canonical mode of regulation. Here, we report a novel activation mechanism for p38α. We show that Arg49 and Arg149 of p38α are methylated by protein arginine methyltransferase 1 (PRMT1). The non-methylation mutations of Lys49/Lys149 abolish the promotive effect of p38α on erythroid differentiation. MAPK kinase 3 (MKK3) is identified as the major p38α upstream kinase and MKK3-mediated activation of the R49/149K mutant p38α is greatly reduced. This is due to a profound reduction in the interaction of p38α and MKK3. PRMT1 can enhance both the methylation level of p38α and its interaction with MKK3. However, the phosphorylation of p38α by MKK3 is not a prerequisite for methylation. MAPK-activated protein kinase 2 (MAPKAPK2) is identified as a p38α downstream effector in the PRMT1-mediated promotion of erythroid differentiation. The interaction of MAPKAPK2 with p38α is also significantly reduced in the R49/149K mutant. Together, this study unveils a novel regulatory mechanism of p38α activation via protein arginine methylation on R49/R149 by PRMT1, which impacts partner interaction and thus promotes erythroid differentiation. This study provides a new insight into the complexity of the regulation of the versatile p38α signaling and suggests new directions in intervening p38α signaling.
2-(3-Hydroxy-5-phosphonooxymethyl-2-methyl-4-pyridyl)-1,3-thiazolidine-4-carboxylic Acid, Novel Metabolite of Pyridoxal 5′-Phosphate and Cysteine Is Present in Human Plasma—Chromatographic Investigations
Authors: Piechocka Justyna , Wrońska Monika , Głowacka Iwona E. , Głowacki Rafał
Published: May 2020

It is well-established that aminothiols, to which cysteine (Cys) belongs, are highly reactive towards aldehydes in an aqueous environment, forming substituted thiazolidine carboxylic acids. This report provides evidence that formation of the product containing a thiazolidine ring through non-enzymatic condensation of Cys and an active form of vitamin B6 pyridoxal 5′-phosphate (PLP) occurs in vivo in humans. To prove this point, a new method, based on a gas chromatography coupled with mass spectrometry (GC-MS), has been designed to identify and quantify Cys and PLP adduct, 2-(3-hydroxy-5-phosphonooxymethyl-2-methyl-4-pyridyl)-1,3-thiazolidine-4-carboxylic acid (HPPTCA) in human plasma. The GC-MS assay relies on sample deproteinization by ultrafiltration over cut-off membranes and preconcentration by drying under vacuum, followed by treatment of the residue with derivatization mixture containing anhydrous pyridine, N-trimethylsilyl-N-methyl trifluoroacetamide (MSTFA) and trimethylchlorosilane (TMCS). The method quantifies HPPTCA in a linear range from 1 to 20 µmol L−1, where the lowest standard on the calibration curve refers to the limit of quantification (LOQ). The validity of the method was demonstrated. Furthermore, the method was successfully applied to plasma samples donated by apparently healthy volunteers and breast cancer patients. The GC-MS assay provides a new tool that will hopefully facilitate studies on the role of HPPTCA in living systems.
The Zinc-Finger Domain Containing Protein ZC4H2 Interacts with TRPV4, Enhancing Channel Activity and Turnover at the Plasma Membrane
Authors: Vangeel Laura , Janssens Annelies , Lemmens Irma , Lievens Sam , Tavernier Jan , Voets Thomas
Published: May 2020

The Ca2+-permeable Transient Receptor Potential channel vanilloid subfamily member 4 (TRPV4) is involved in a broad range of physiological processes, including the regulation of systemic osmotic pressure, bone resorption, vascular tone, and bladder function. Mutations in the TRPV4 gene are the cause of a spectrum of inherited diseases (or TRPV4-pathies), which include skeletal dysplasias, arthropathies, and neuropathies. There is little understanding of the pathophysiological mechanisms underlying these variable disease phenotypes, but it has been hypothesized that disease-causing mutations affect interaction with regulatory proteins. Here, we performed a mammalian protein–protein interaction trap (MAPPIT) screen to identify proteins that interact with the cytosolic N terminus of human TRPV4, a region containing the majority of disease-causing mutations. We discovered the zinc-finger domain-containing protein ZC4H2 as a TRPV4-interacting protein. In heterologous expression experiments, we found that ZC4H2 increases both the basal activity of human TRPV4 as well as Ca2+ responses evoked by ligands or hypotonic cell swelling. Using total internal reflection fluorescence (TIRF) microscopy, we further showed that ZC4H2 accelerates TRPV4 turnover at the plasma membrane. Overall, these data demonstrate that ZC4H2 is a positive modulator of TRPV4, and suggest a link between TRPV4 and ZC4H2-associated rare disorders, which have several neuromuscular symptoms in common with TRPV4-pathies.
Serum Exosomal miRNAs for Grading Hepatic Fibrosis Due to Schistosomiasis
Authors: Cai Pengfei , Mu Yi , Olveda Remigio M. , Ross Allen G. , Olveda David U. , McManus Donald P.
Published: May 2020

Chronic infection with Schistosoma japonicum or Schistosoma mansoni results in hepatic fibrosis of the human host. The staging of fibrosis is crucial for prognosis and to determine the need for treatment of patients with schistosomiasis. This study aimed to determine whether there is a correlation between the levels of serum exosomal micro-ribonucleic acids (miRNAs) (exomiRs) and fibrosis progression in schistosomiasis. Reference gene (RG) validation was initially carried out for the analysis of serum exomiRs expression in staging liver fibrosis caused by schistosome infection. The expression levels of liver fibrosis-associated exomiRs in serum were determined in a murine schistosomiasis model and in a cohort of Filipino schistosomiasis japonica patients (n = 104) with different liver fibrosis grades. Of twelve RG candidates validated, miR-103a-3p and miR-425-5p were determined to be the most stable genes in the murine schistosomiasis model and subjects from the schistosomiasis-endemic area, respectively. The temporal expression profiles of nine fibrosis-associated serum exomiRs, as well as their correlations with the liver pathologies, were determined in C57BL/6 mice during S. japonicum infection. The serum levels of three exomiRs (miR-92a-3p, miR-146a-5p and miR-532-5p) were able to distinguish subjects with fibrosis grades I-III from those with no fibrosis, but only the serum level of exosomal miR-146a-5p showed potential for distinguishing patients with mild (grades 0–I) versus severe fibrosis (grades II–III). The current data imply that serum exomiRs can be a supplementary tool for grading liver fibrosis in hepatosplenic schistosomiasis with moderate accuracy.
Polyphenols with Anti-Amyloid β Aggregation Show Potential Risk of Toxicity Via Pro-Oxidant Properties
Authors: Kobayashi Hatasu , Murata Mariko , Kawanishi Shosuke , Oikawa Shinji
Published: May 2020

Alzheimer’s disease (AD) is the most common form of dementia among older people. Amyloid β (Aβ) aggregation has been the focus for a therapeutic target for the treatment of AD. Naturally occurring polyphenols have an inhibitory effect on Aβ aggregation and have attracted a lot of attention for the development of treatment strategies which could mitigate the symptoms of AD. However, considerable evidence has shown that the pro-oxidant mechanisms of polyphenols could have a deleterious effect. Our group has established an assay system to evaluate the pro-oxidant characteristics of chemical compounds, based on their reactivity with DNA. In this review, we have summarized the anti-Aβ aggregation and pro-oxidant properties of polyphenols. These findings could contribute to understanding the mechanism underlying the potential risk of polyphenols. We would like to emphasize the importance of assessing the pro-oxidant properties of polyphenols from a safety point of view.
Carbonic Anhydrase Sensitivity to Pesticides: Perspectives for Biomarker Development
Authors: Lionetto Maria Giulia , Caricato Roberto , Giordano Maria Elena
Published: May 2020

Carbonic anhydrase (CA) is a widespread metalloenzyme playing a pivotal role in several physiological processes. Many studies have demonstrated the in vitro and in vivo sensitivity of CA to the exposure to several classes of pesticides in both humans and wildlife. This review aims to analyze and to discuss the literature available in this field, providing a comprehensive view useful to foresee perspectives for the development of novel CA-based pesticide biomarkers. The analysis of the available data highlighted the ability of several pesticide molecules to interact directly with the enzyme in humans and wildlife and to inhibit CA activity in vitro and in vivo, with possible alterations of key physiological functions. The analysis disclosed key areas of further research and, at the same time, identified some perspectives for the development of novel CA-based sensitive biomarkers to pesticide exposure, suitable to be used in several fields from human biomonitoring in occupational and environmental medicine to environmental monitoring on non-target species.
Safety and Effectiveness of Copaiba Oleoresin (C. reticulata Ducke) on Inflammation and Tissue Repair of Oral Wounds in Rats
Authors: Alvarenga María Olimpia Paz , Bittencourt Leonardo Oliveira , Mendes Paulo Fernando Santos , Ribeiro Julia Turra , Lameira Osmar Alves , Monteiro Marta Chagas , Barboza Carlos Augusto Galvão , Martins Manoela Domingues , Lima Rafael Rodrigues
Published: May 2020

In traditional communities of the Brazilian Amazon, the copaiba oleoresin (C. reticulata Ducke) is widely known for its therapeutic activity, especially its wound healing and anti-inflammatory actions. Our study aimed to evaluate these effects in oral lesions and the safety of the dosage proposed. A punch biopsy wound was induced on the ventral surface of the tongue of forty-five male Wistar rats under anesthesia. Animals were randomly allocated to one of three groups based on the treatment: control, corticoid and copaiba. A daily dose of each treatment and vehicle was administrated by oral gavage for three consecutive days. Sample collections took place on the third, seventh and 15th days post-wounding for clinical and histopathological analyses. Blood was collected on the third and seventh days for kidneys and liver function tests. Semi-quantitative analyses were performed based on scores of inflammation and reepithelization. Tissue collagen deposition was detected by PicroSirius red staining. Copaiba-treated wounds revealed a smaller wound area, decreased of acute inflammatory reaction and enhanced reepithelization. The levels of kidney and liver function tests did not reveal presence of damage post-treatments. Our findings suggest that copaiba oleoresin is a safe and effective alternative therapy for inflammation and tissue repair of oral wounds in this animal model.
C-Fiber Loss as a Possible Cause of Neuropathic Pain in Schwannomatosis
Authors: Farschtschi Said , Mainka Tina , Glatzel Markus , Hannekum Anna-Lena , Hauck Michael , Gelderblom Mathias , Hagel Christian , Friedrich Reinhard , Schuhmann Martin , Schulz Alexander , Morrison Helen , Kehrer-Sawatzki Hildegard , Luhmann Jan , Gerloff Christian , Bendszus Martin , Bäumer Philipp , Mautner Victor-Felix
Published: May 2020

Schwannomatosis is the third form of neurofibromatosis and characterized by the occurrence of multiple schwannomas. The most prominent symptom is chronic pain. We aimed to test whether pain in schwannomatosis might be caused by small-fiber neuropathy. Twenty patients with schwannomatosis underwent neurological examination and nerve conduction studies. Levels of pain perception as well as anxiety and depression were assessed by established questionnaires. Quantitative sensory testing (QST) and laser-evoked potentials (LEP) were performed on patients and controls. Whole-body magnetic resonance imaging (wbMRI) and magnetic resonance neurography (MRN) were performed to quantify tumors and fascicular nerve lesions; skin biopsies were performed to determine intra-epidermal nerve fiber density (IENFD). All patients suffered from chronic pain without further neurological deficits. The questionnaires indicated neuropathic symptoms with significant impact on quality of life. Peripheral nerve tumors were detected in all patients by wbMRI. MRN showed additional multiple fascicular nerve lesions in 16/18 patients. LEP showed significant faster latencies compared to normal controls. Finally, IENFD was significantly reduced in 13/14 patients. Our study therefore indicates the presence of small-fiber neuropathy, predominantly of unmyelinated C-fibers. Fascicular nerve lesions are characteristic disease features that are associated with faster LEP latencies and decreased IENFD. Together these methods may facilitate differential diagnosis of schwannomatosis.
Long-Term Lead Exposure Since Adolescence Causes Proteomic and Morphological Alterations in the Cerebellum Associated with Motor Deficits in Adult Rats
Authors: Leão Luana Ketlen Reis , Bittencourt Leonardo Oliveira , Oliveira Ana Carolina , Nascimento Priscila Cunha , Miranda Giza Hellen Nonato , Ferreira Railson Oliveira , Nabiça Mariane , Dantas Kelly , Dionizio Aline , Cartágenes Sabrina , Buzalaf Marília Afonso Rabelo , Crespo-Lopez Maria Elena , Maia Cristiane S F , Lima Rafael Rodrigues
Published: May 2020

Lead (Pb) is an environmental contaminant that presents a high risk for human health. We aimed to investigate the possible alterations triggered by the exposure to Pb acetate for a long period in motor performance and the possible relationship with biochemical, proteomic and morphological alterations in the cerebellum of rats. Male Wistar rats were exposed for 55 days, at 50 mg/Kg of Pb acetate, and the control animals received distilled water. Open field (OF) and rotarod tests; biochemistry parameters (MDA and nitrite); staining/immunostaining of Purkinje cells (PC), mature neurons (MN), myelin sheath (MS) and synaptic vesicles (SYN) and proteomic profile were analyzed. Pb deposition on the cerebellum area and this study drove to exploratory and locomotion deficits and a decrease in the number of PC, MN, SYN and MS staining/immunostaining. The levels of MDA and nitrite remained unchanged. The proteomic profile showed alterations in proteins responsible for neurotransmitters release, as well as receptor function and second messengers signaling, and also proteins involved in the process of apoptosis. Thus, we conclude that the long-term exposure to low Pb dose promoted locomotion and histological tracings, associated with alterations in the process of cell signaling, as well as death by apoptosis.
Family Caregiver Strain and Challenges When Caring for Orthopedic Patients: A Systematic Review
Authors: Longo Umile Giuseppe , Matarese Maria , Arcangeli Valeria , Alciati Viviana , Candela Vincenzo , Facchinetti Gabriella , Marchetti Anna , De Marinis Maria Grazia , Denaro Vincenzo
Published: May 2020

Background: Caregivers represent the core of patients’ care in hospital structures, in the process of care and self-care after discharge. We aim to identify the factors that affect the strain of caring for orthopedic patients and how these factors are related to the quality of life of caregivers. We also want to evaluate the role of caregivers in orthopedic disease, focusing attention on the patient–caregiver dyad. Methods: A comprehensive search on PubMed, Cochrane, CINAHL and Embase databases was conducted. This review was reported following PRISMA statement guidance. Studies were selected, according to inclusion and exclusion criteria, about patient–caregiver dyads. For quality assessment, we used the MINORS and the Cochrane Risk of BIAS assessment tool. Results: 28 studies were included in the systematic review; in these studies, 3034 dyads were analyzed. Caregivers were not always able to bear the difficulties of care. An improvement in strain was observed after behavioral interventions from health-care team members; Conclusions: The role of the caregiver can lead to a deterioration of physical, cognitive and mental conditions. The use of behavioral interventions increased quality of life, reducing the strain in caregivers of orthopedic patients. For this reason, it is important to consider the impact that orthopedic disease has on the strain of the caregiver and to address this topic.
Initial Steps for the Development of a Phage-Mediated Gene Replacement Therapy Using CRISPR-Cas9 Technology
Authors: Yang Zhou Jordi , Suwan Keittisak , Hajitou Amin
Published: May 2020

p53 gene (TP53) replacement therapy has shown promising results in cancer gene therapy. However, it has been hampered, mostly because of the gene delivery vector of choice. CRISPR-Cas9 technology (clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9) can knock out the mutated TP53 (mutTP53), but due to its large size, many viral vectors are not suitable or require implemented strategies that lower the therapeutic efficiency. Here, we introduced a bacteriophage or phage-based vector with the ability to target cancer cells and aimed to investigate the feasibility of using this vector to deliver CRISPR-Cas9 transgene in human lung adenocarcinoma cells. First, we produced a tumour-targeted bacteriophage carrying a CRISPR-Cas9 transgene cassette. Next, we investigated any negative impact on vector titers via quantitative polymerase chain reaction (qPCR) and colony-forming agar plate. Last, we combined Western blot analysis and immunofluorescence staining to prove cell transduction in vitro. We showed that the tumour-targeted bacteriophage can package a large-size vector genome, ~10 kb, containing the CRISPR-Cas9 sequence without any negative impact on the active or total number of bacteriophage particles. Then, we detected expression of the Cas9 in human lung adenocarcinoma cells in a targeted and efficient manner. Finally, we proved loss of p53 protein expression when a p53 gRNA was incorporated into the CRISPR-Cas9 phage DNA construct. These proof-of-concept findings support the use of engineered bacteriophage for TP53 replacement therapy in lung cancer.
Epigenetic Regulation of Alternative mRNA Splicing in Dilated Cardiomyopathy
Authors: Gi Weng-Tein , Haas Jan , Sedaghat-Hamedani Farbod , Kayvanpour Elham , Tappu Rewati , Lehmann David Hermann , Shirvani Samani Omid , Wisdom Michael , Keller Andreas , Katus Hugo A. , Meder Benjamin
Published: May 2020

In recent years, the genetic architecture of dilated cardiomyopathy (DCM) has been more thoroughly elucidated. However, there is still insufficient knowledge on the modifiers and regulatory principles that lead to the failure of myocardial function. The current study investigates the association of epigenome-wide DNA methylation and alternative splicing, both of which are important regulatory principles in DCM. We analyzed screening and replication cohorts of cases and controls and identified distinct transcriptomic patterns in the myocardium that differ significantly, and we identified a strong association of intronic DNA methylation and flanking exons usage (p < 2 × 10−16). By combining differential exon usage (DEU) and differential methylation regions (DMR), we found a significant change of regulation in important sarcomeric and other DCM-associated pathways. Interestingly, inverse regulation of Titin antisense non-coding RNA transcript splicing and DNA methylation of a locus reciprocal to TTN substantiate these findings and indicate an additional role for non-protein-coding transcripts. In summary, this study highlights for the first time the close interrelationship between genetic imprinting by DNA methylation and the transport of this epigenetic information towards the dynamic mRNA splicing landscape. This expands our knowledge of the genome–environment interaction in DCM besides simple gene expression regulation.