bluuebunny/crossref_metadata_2025 huggingface dataset:
TREAT-B Algorithm for Treatment Eligibility Among Chronically Infected Hepatitis B Virus Persons in a Low and a High Endemic Region: A Potential Strategy Towards Virus Elimination by 2030
Authors: Kafeero Hussein Mukasa , Ndagire Dorothy , Ocama Ponsiano , Kato Charles Drago , Wampande Eddie , Kajumbula Henry , Kateete David Patrick , Walusansa Abdul , Kudamba Ali , Ssenku Jamilu E. , Sendagire Hakim
Published: April 2022

Background

Little is known about treatment eligibility in Africa for the hepatitis B virus (TREAT-B) algorithm. We investigated the treatment eligibility among the HBV chronically infected patients in a low and a high endemic region using the TREAT-B algorithm.

Methods

We recruited 227 treatment-naïve HBV-infected hospital attendees from the low and high HBV endemic regions. We assessed the treatment eligibility by testing for HBeAg serostatus and ALT levels. Socio-demographic data were collected with a structured questionnaire. The accessory correlates of treatment eligibility (AST, ALP, ALB, GGT, and TBIL) and the socio-demographic factors were analyzed by both univariate and multinomial logistic regression using the SPSS and Medcalc. The analysis was done at 95% CI and a p < 0.05 was considered statistically significant.

Results

Overall, 56.8% of the participants qualified for treatment at TREAT-B cutoffs of ≥2, with those from the low endemic region (90, 69.8%) having significantly higher treatment eligibility indication than those from the high endemic region (p < 0.05). Alcohol use and household contact with an HBV-infected person were independent socio-demographic factors significantly associated with treatment eligibility for both low and high endemic regions (p < 0.05). However, birth place was only indicated for treatment eligibility among the high endemic participants (p < 0.05). AST, GGT, and total bilirubin were the liver-related parameters significantly associated with treatment eligibility (p < 0.05), with GGT and AST being significantly elevated among the eligible low endemic dwellers compared to high endemic dwellers (p < 0.05).

Conclusion

Using TREAT-B algorithm can be a plausible alternative to the orthodox methods to specify treatment eligibility with the potential to scale up interventions targeting HBV management and elimination.
PD-L1 Testing in Urothelial Carcinoma: Analysis of a Series of 1401 Cases Using Both the 22C3 and SP142 Assays
Authors: Evans Harriet , O’Sullivan Brendan , Hughes Frances , Charles Kathryn , Robertson Lee , Taniere Philippe , Diaz-Cano Salvador
Published: April 2022

Immune checkpoint blockade (ICB) drugs are a novel, effective treatment for advanced urothelial carcinoma. Worldwide, several different ICB drugs are approved, each developed and clinically validated with a specific PD-L1 compound diagnostic assay. As a result, PD-L1 testing workflows in routine practice are complex: requiring multiple assays across two platforms, with each assay having a different method of interpretation. Our service tested 1,401 urothelial carcinoma cases for PD-L1 expression, using both the 22C3 PharmDx assay (required prior to Pembrolizumab therapy) and SP142 assay (required prior to Atezolizumab therapy). Of the 1,401 cases tested, 621 cases (44%) were tested with both the 22C3 PharmDx and SP142 assays, 492 cases (35%) with 22C3 PharmDx only, and 288 cases (21%) with SP142 only. Each assay was used and interpreted according to the manufacturer’s guidelines. The rate of positivity we observed was 26% with the 22C3 assay and 31% with the SP142 assay, similar to the pre-licensing studies for both drugs. The discrepancy observed between the assays was 11%, which reinforces the requirement for utilisation of the correct assay for each agent, and limits potential cross-utility of assays. This aspect must be considered when setting up a PD-L1 testing strategy in laboratories where both Pembrolizumab and Atezolizumab are available for the treatment of urothelial carcinoma but also has broader implications for testing of other cancers where multiple ICB drugs and their respective assays are approved.
Animal Abuse as an Indicator of Domestic Violence: One Health, One Welfare Approach
Authors: Mota-Rojas Daniel , Monsalve Stefany , Lezama-García Karina , Mora-Medina Patricia , Domínguez-Oliva Adriana , Ramírez-Necoechea Ramiro , Garcia Rita de Cassia Maria
Published: April 2022

For years now, the importance of animal cruelty has been gaining recognition in the industrialized cities of the West. Animal cruelty encompasses any act that causes a non-human animal unnecessary pain or suffering, including negligence, abandonment, abuse, torture, bestiality, and even theriocide. This represents a red flag for society as a whole because people who commit such acts can escalate violence and direct it to other individuals. Animal cruelty and interpersonal violence—as well as other socially undesirable conduct such as bullying, antisocial personality disorder, rape, and serial murder—are closely related, so timely diagnoses of either one can help prevent acts of aggression. It is necessary, therefore, to analyze and try to understand whether there are early indicators that may help identify potentially violent individuals. It is well known that kids from homes with actual violence in their homes show a high tendency to reproduce such behaviors with both animals and other people. In conclusion, much research and rethinking of the importance of the veterinarian in detecting animal abuse and cruelty is needed to help detect and prevent cases of interpersonal violence that may arise over time.
Applicability of Weighting Method as Measure for Existing Manholes against Uplifting during Liquefaction
Authors: Nakao Koki , Yamaguchi Hiroo , Hoshino Shigefumi , Inazumi Shinya
Published: April 2022

It has been confirmed that manholes may protrude from the road surface due to the liquefaction of the ground during a large earthquake. When the uplifting of a manhole occurs, it may bring about not only dysfunctions, such as the loss of the ability of sewage to flow, but also displacements, such as that of the pipeline connected to the manhole, which makes drainage difficult. However, effective and economic measures against the uplifting of existing manholes are still in the developmental stages. This study proposes a weighting method that increases the self-weight of the manhole as a countermeasure against the uplifting of manholes during an earthquake. The applicability of this weighting method will be clarified by conducting case studies from the viewpoint of the safety factor for manhole uplifting and the quantity of manhole uplifting. As a result, it is shown that, regardless of the shape of the manhole and the groundwater level, the risk of manhole uplifting during liquefaction can be reduced by implementing this weighting method.
Artificial Intelligence in the Management of Barrett’s Esophagus and Early Esophageal Adenocarcinoma
Authors: Dumoulin Franz Ludwig , Rodriguez-Monaco Fabian Dario , Ebigbo Alanna , Steinbrück Ingo
Published: April 2022

Esophageal adenocarcinoma is increasing in incidence and is the most common subtype of esophageal cancer in Western societies. The stepwise progression of Barrett´s metaplasia to high-grade dysplasia and invasive adenocarcinoma provides an opportunity for screening and surveillance. There are important unresolved issues, which include (i) refining the definition of the screening population in order to avoid unnecessary invasive diagnostics, (ii) a more precise prediction of the (very heterogeneous) individual progression risk from metaplasia to invasive cancer in order to better tailor surveillance recommendations, (iii) improvement of the quality of endoscopy in order to reduce the high miss rate for early neoplastic lesions, and (iv) support for the diagnosis of tumor infiltration depth in order to guide treatment decisions. Artificial intelligence (AI) systems might be useful as a support to better solve the above-mentioned issues.
N-Terminus-Mediated Solution Structure of Dimerization Domain of PRC1
Authors: Tan Fei , Xu Jin
Published: April 2022

Microtubule-associated proteins (MAPs) are essential for the accurate division of a cell into two daughter cells. These proteins target specific microtubules to be incorporated into the spindle midzone, which comprises a special array of microtubules that initiate cytokinesis during anaphase. A representative member of the MAPs is Protein Regulator of Cytokinesis 1 (PRC1), which self-multimerizes to cross-link microtubules, the malfunction of which might result in cancerous cells. The importance of PRC1 multimerization makes it a popular target for structural studies. The available crystal structure of PRC1 has low resolution (>3 Å) and accuracy, limiting a better understanding of the structure-related functions of PRC1. Therefore, we used NMR spectroscopy to better determine the structure of the dimerization domain of PRC1. The NMR structure shows that the PRC1 N terminus is crucial to the overall structure integrity, but the crystal structure bespeaks otherwise. We systematically addressed the role of the N terminus by generating a series of mutants in which N-terminal residues methionine (Met1) and arginine (Arg2) were either deleted, extended or substituted with other rationally selected amino acids. Each mutant was subsequently analyzed by NMR spectroscopy or fluorescence thermal shift assays for its structural or thermal stability; we found that N-terminal perturbations indeed affected the overall protein structure and that the solution structure better reflects the conformation of PRC1 under solution conditions. These results reveal that the structure of PRC1 is governed by its N terminus through hydrophobic interactions with other core residues, such hitherto unidentified N-terminal conformations might shed light on the structure–function relationships of PRC1 or other proteins. Therefore, our study is of major importance in terms of identifying a novel structural feature and can further the progress of protein folding and protein engineering.
Mechanical and Conductive Performance of Aged 6xxx Aluminum Alloy during Rotary Swaging
Authors: Jin Hongmei , Guan Renguo , Tie Di
Published: April 2022

Thermomechanical treatment consisting of heat treatment and deformation is an effective processing route for precipitation-hardened 6xxx alloy (Al-Mg-Si-Cu system), and precipitates and dislocations produced during the process can significantly change its mechanical and conductive performance. We therefore investigated the microstructural evolution of precipitates in a representative 6xxx alloy during thermomechanical treatment. When the precipitates encountered the accumulated dislocations, the precipitates were bent and broken into dispersed smaller particles. The strength of the alloy was significantly improved by the proliferation of dislocations and precipitates and desired electrical conductivity was obtained as well. Our results prove that peak aging plus cold rotary swaging is an efficient processing route for simultaneously improving the mechanical and conductive performance of 6xxx alloy.
DNA Barcoding of Fresh and Historical Collections of Lichen-Forming Basidiomycetes in the Genera Cora and Corella (Agaricales: Hygrophoraceae): A Success Story?
Authors: Dal Forno Manuela , Lawrey James D. , Moncada Bibiana , Bungartz Frank , Grube Martin , Schuettpelz Eric , Lücking Robert
Published: April 2022

Lichens collected worldwide for centuries have resulted in millions of specimens deposited in herbaria that offer the potential to assess species boundaries, phenotypic diversification, ecology, and distribution. The application of molecular approaches to historical collections has been limited due to DNA fragmentation, but high-throughput sequencing offers an opportunity to overcome this barrier. Here, we combined a large dataset of ITS sequences from recently collected material and historical collections, obtained through Sanger, 454, or Illumina Sequencing, to test the performance of ITS barcoding in two genera of lichenized Basidiomycota: Cora and Corella. We attempted to generate new sequence data for 62 fresh specimens (from 2016) and 274 historical collections (collected between 1888 and 1998), for a final dataset of 1325 sequences. We compared various quantitative approaches to delimit species (GMYC, bPTP, ASAP, ABGD) and tested the resolution and accuracy of the ITS fungal barcoding marker by comparison with a six-marker dataset. Finally, we quantitatively compared phylogenetic and phenotypic species delimitation for 87 selected Cora species that have been formally described. Our HTS approach successfully generated ITS sequences for 76% of the historical collections, and our results show that an integrative approach is the gold-standard for understanding diversity in this group.
Electroosmosis-Optimized Thermal Model for Peristaltic Transportation of Thermally Radiative Magnetized Liquid with Nonlinear Convection
Authors: Akbar Yasir , Alotaibi Hammad
Published: April 2022

The present study addresses the heat transfer efficiency and entropy production of electrically conducting kerosene-based liquid led by the combined impact of electroosmosis and peristalsis mechanisms. Effects of nonlinear mixed convection heat transfer, temperature-dependent viscosity, radiative heat flux, electric and magnetic fields, porous medium, heat sink/source, viscous dissipation, and Joule heating are presented. The Debye–Huckel linearization approximation is employed in the electrohydrodynamic problem. Mathematical modeling is conducted within the limitations of δ << 1 and Re → 0. Coupled differential equations after implementing a lubrication approach are numerically solved. The essential characteristics of the production of entropy, the factors influencing it, and the characteristics of heat and fluid in relation to various physical parameters are graphically evaluated by assigning them a growing list of numeric values. This analysis reveals that heat transfer enhances by enhancing nonlinear convection and Joule heating parameters. The irreversibility analysis ensures that the minimization of entropy generation is observed when the parameters of viscosity and radiation are held under control. Fluid velocity can be regulated by adjusting the Helmholtz–Smoluchowski velocity and magnetic field strength.
A Toss without a Coin: Information, Discontinuity, and Mathematics in Quantum Theory
Authors: Plotnitsky Arkady
Published: April 2022

The article argues that—at least in certain interpretations, such as the one assumed in this article under the heading of “reality without realism”—the quantum-theoretical situation appears as follows: While—in terms of probabilistic predictions—connected to and connecting the information obtained in quantum phenomena, the mathematics of quantum theory (QM or QFT), which is continuous, does not represent and is discontinuous with both the emergence of quantum phenomena and the physics of these phenomena, phenomena that are physically discontinuous with each other as well. These phenomena, and thus this information, are described by classical physics. All actually available information (in the mathematical sense of information theory) is classical: it is composed of units, such as bits, that are—or are contained in—entities described by classical physics. On the other hand, classical physics cannot predict this information when it is created, as manifested in measuring instruments, in quantum experiments, while quantum theory can. In this epistemological sense, this information is quantum. The article designates the discontinuity between quantum theory and the emergence of quantum phenomena the “Heisenberg discontinuity”, because it was introduced by W. Heisenberg along with QM, and the discontinuity between QM or QFT and the classical physics of quantum phenomena, the “Bohr discontinuity”, because it was introduced as part of Bohr’s interpretation of quantum phenomena and QM, under the assumption of Heisenberg discontinuity. Combining both discontinuities precludes QM or QFT from being connected to either physical reality, that ultimately responsible for quantum phenomena or that of these phenomena themselves, other than by means of probabilistic predictions concerning the information, classical in character, contained in quantum phenomena. The nature of quantum information is, in this view, defined by this situation. A major implication, discussed in the Conclusion, is the existence and arguably the necessity of two—classical and quantum—or with relativity, three and possibly more essentially different theories in fundamental physics.
A Pragmatic Ensemble Strategy for Missing Values Imputation in Health Records
Authors: Batra Shivani , Khurana Rohan , Khan Mohammad Zubair , Boulila Wadii , Koubaa Anis , Srivastava Prakash
Published: April 2022

Pristine and trustworthy data are required for efficient computer modelling for medical decision-making, yet data in medical care is frequently missing. As a result, missing values may occur not just in training data but also in testing data that might contain a single undiagnosed episode or a participant. This study evaluates different imputation and regression procedures identified based on regressor performance and computational expense to fix the issues of missing values in both training and testing datasets. In the context of healthcare, several procedures are introduced for dealing with missing values. However, there is still a discussion concerning which imputation strategies are better in specific cases. This research proposes an ensemble imputation model that is educated to use a combination of simple mean imputation, k-nearest neighbour imputation, and iterative imputation methods, and then leverages them in a manner where the ideal imputation strategy is opted among them based on attribute correlations on missing value features. We introduce a unique Ensemble Strategy for Missing Value to analyse healthcare data with considerable missing values to identify unbiased and accurate prediction statistical modelling. The performance metrics have been generated using the eXtreme gradient boosting regressor, random forest regressor, and support vector regressor. The current study uses real-world healthcare data to conduct experiments and simulations of data with varying feature-wise missing frequencies indicating that the proposed technique surpasses standard missing value imputation approaches as well as the approach of dropping records holding missing values in terms of accuracy.
Latency-Optimized Design of Data Bus Inversion
Authors: Pae Sung-il , Kwon Kon-Woo
Published: April 2022

This paper proposes two new encoders for data bus inversion (DBI), which conventionally uses a majority voter to pick a data representation that minimizes switching activities and thus reduces the corresponding energy consumption. The new encoders employ simpler approximate voters comprising only two gate levels, which improve latency more than twice while still achieving switching activity savings by 9% and 11%, respectively. Although the proposed voters are not always accurate, the errors in the voters do not affect the correctness of data movement. We report various metrics, including latencies, areas, and operating powers, regarding five different designs, two proposed designs along with three conventional designs, based on 65-nm process implementations.
FMECA Application in Tomotherapy: Comparison between Classic and Fuzzy Methodologies
Authors: Giardina Mariarosa , Tomarchio Elio , Buffa Pietro , Palagonia Maurizio , Veronese Ivan , Cantone Marie Claire
Published: April 2022

Accident analysis in radiotherapy highlighted the need to increase quality assurance (QA) programs by the identification of failures/errors with very low probability (rare event) but very severe consequences. In this field, a Failure Mode, Effects and Criticality Analysis (FMECA) technique, used in various industrial processes to rank critical events, has been met with much interest. The literature describes different FMECA methods; however, it is necessary to understand if these tools are incisive and effective in the healthcare sector. In this work, comparisons of FMECA methodologies in the risk assessment of patients undergoing treatments performed with helical tomotherapy are reported. Failure modes identified for the phases “treatment planning” and “treatment execution” are classified using the Risk Priority Number (RPN) index. Differences and similarities in the classification of failures/errors of the examined FMECA approaches are highlighted.
Improvement of Oil Valorization Extracted from Fish By-Products Using a Handheld near Infrared Spectrometer Coupled with Chemometrics
Authors: Nieto-Ortega Sonia , Olabarrieta Idoia , Saitua Eduardo , Arana Gorka , Foti Giuseppe , Melado-Herreros Ángela
Published: April 2022

A handheld near infrared (NIR) spectrometer was used for on-site determination of the fatty acids (FAs) composition of industrial fish oils from fish by-products. Partial least square regression (PLSR) models were developed to correlate NIR spectra with the percentage of saturated fatty acids (SFAs), monounsaturated fatty acids (MUFAs), polyunsaturated fatty acids (PUFAs) and, among them, omega-3 (ω-3) and omega-6 (ω-6) FAs. In a first step, the data were divided into calibration validation datasets, obtaining good results regarding R2 values, root mean square error of prediction (RMSEP) and bias. In a second step, all these data were used to create a new calibration, which was uploaded to the handheld device and tested with an external validation set in real time. Evaluation of the external test set for SFAs, MUFAs, PUFAs and ω-3 models showed promising results, with R2 values of 0.98, 0.97, 0.97 and 0.99; RMSEP (%) of 0.94, 1.71, 1.11 and 0.98; and bias (%) values of −0.78, −0.12, −0.80 and −0.67, respectively. However, although ω-6 models achieved a good R2 value (0.95), the obtained RMSEP was considered high (2.08%), and the bias was not acceptable (−1.76%). This was corrected by applying bias and slope correction (BSC), obtaining acceptable values of R2 (0.95), RMSEP (1.09%) and bias (−0.05%). This work goes a step further in the technology readiness level (TRL) of handheld NIR sensor solutions for the fish by-product recovery industry.
Targeted Cancer Therapy via pH-Functionalized Nanoparticles: A Scoping Review of Methods and Outcomes
Authors: Morarasu Stefan , Morarasu Bianca Codrina , Ghiarasim Razvan , Coroaba Adina , Tiron Crina , Iliescu Radu , Dimofte Gabriel-Mihail
Published: April 2022

(1) Background: In recent years, several studies have described various and heterogenous methods to sensitize nanoparticles (NPs) to pH changes; therefore, in this current scoping review, we aimed to map current protocols for pH functionalization of NPs and analyze the outcomes of drug-loaded pH-functionalized NPs (pH-NPs) when delivered in vivo in tumoral tissue. (2) Methods: A systematic search of the PubMed database was performed for all published studies relating to in vivo models of anti-tumor drug delivery via pH-responsive NPs. Data on the type of NPs, the pH sensitization method, the in vivo model, the tumor cell line, the type and name of drug for targeted therapy, the type of in vivo imaging, and the method of delivery and outcomes were extracted in a separate database. (3) Results: One hundred and twenty eligible manuscripts were included. Interestingly, 45.8% of studies (n = 55) used polymers to construct nanoparticles, while others used other types, i.e., mesoporous silica (n = 15), metal (n = 8), lipids (n = 12), etc. The mean acidic pH value used in the current literature is 5.7. When exposed to in vitro acidic environment, without exception, pH-NPs released drugs inversely proportional to the pH value. pH-NPs showed an increase in tumor regression compared to controls, suggesting better targeted drug release. (4) Conclusions: pH-NPs were shown to improve drug delivery and enhance antitumoral effects in various experimental malignant cell lines.
Changes in Motor Performance and BMI of Primary School Children over Time–Influence of the COVID-19 Confinement and Social Burden
Authors: Wessely Stefanie , Ferrari Nina , Friesen David , Grauduszus Martin , Klaudius Marlen , Joisten Christine
Published: April 2022

During the COVID-19 pandemic, physical inactivity and being overweight increased in children, especially those from lower socioeconomic backgrounds. Additionally, a decrease in motor performance has been increasingly reported; however, there is a lack of meaningful data on the relationship between these findings and socioeconomic status. Therefore, we examined the BMI, BMI z score, the prevalence of overweight/obesity, and motor performance (based on lateral jumping, the standing long jump, and the 6-min run) among three cohorts from timelines before (2016) and during the pandemic (2020, 2021), using a cohort sequential design. Our analysis revealed that all parameters examined showed a significant negative trend over time. For children with a low social burden, the trend was statistically detectable only for lateral jumping and the 6-min run. For children with a high social burden, the prevalence of overweight/obesity increased from 4.2% to 5.4% between 2016 and 2021. The prevalence of being underweight also increased from 3.4% to 4.3% during this time. Motor performance decreased from 11.3% to 12.3%, except in the standing long jump. Children with a high social burden showed significantly lower results (7.4% to 9.6%). Thus, our data confirm an increase in both measurements of overweight/obesity and underweight, as well as a decrease in motor performance under the pandemic conditions, especially in children from socioeconomically deprived backgrounds. These findings affirm the urgent need for action regarding health promotion measures, especially for children with high social burdens.
Urban Change Detection from Aerial Images Using Convolutional Neural Networks and Transfer Learning
Authors: Fyleris Tautvydas , Kriščiūnas Andrius , Gružauskas Valentas , Čalnerytė Dalia , Barauskas Rimantas
Published: April 2022

Urban change detection is an important part of sustainable urban planning, regional development, and socio-economic analysis, especially in regions with limited access to economic and demographic statistical data. The goal of this research is to create a strategy that enables the extraction of indicators from large-scale orthoimages of different resolution with practically acceptable accuracy after a short training process. Remote sensing data can be used to detect changes in number of buildings, forest areas, and other landscape objects. In this paper, aerial images of a digital raster orthophoto map at scale 1:10,000 of the Republic of Lithuania (ORT10LT) of three periods (2009–2010, 2012–2013, 2015–2017) were analyzed. Because of the developing technologies, the quality of the images differs significantly and should be taken into account while preparing the dataset for training the semantic segmentation model DeepLabv3 with a ResNet50 backbone. In the data preparation step, normalization techniques were used to ensure stability of image quality and contrast. Focal loss for the training metric was selected to deal with the misbalanced dataset. The suggested model training process is based on the transfer learning technique and combines using a model with weights pretrained in ImageNet with learning on coarse and fine-tuning datasets. The coarse dataset consists of images with classes generated automatically from Open Street Map (OSM) data and the fine-tuning dataset was created by manually reviewing the images to ensure that the objects in images match the labels. To highlight the benefits of transfer learning, six different models were trained by combining different steps of the suggested model training process. It is demonstrated that using pretrained weights results in improved performance of the model and the best performance was demonstrated by the model which includes all three steps of the training process (pretrained weights, training on coarse and fine-tuning datasets). Finally, the results obtained with the created machine learning model enable the implementation of different approaches to detect, analyze, and interpret urban changes for policymakers and investors on different levels on a local map, grid, or municipality level.
Absolute Quantification of Nav1.5 Expression by Targeted Mass Spectrometry
Authors: Adams Sarah , Chang Ge , Fouda Mohamed , Kumar Sharwan , Sun Bingyun
Published: April 2022

Nav1.5 is the pore forming α-subunit of the cardiac voltage-gated sodium channel that initiates cardiac action potential and regulates the human heartbeat. A normal level of Nav1.5 is crucial to cardiac function and health. Over- or under-expression of Nav1.5 can cause various cardiac diseases ranging from short PR intervals to Brugada syndromes. An assay that can directly quantify the protein amount in biological samples would be a priori to accurately diagnose and treat Nav1.5-associated cardiac diseases. Due to its large size (>200 KD), multipass transmembrane domains (24 transmembrane passes), and heavy modifications, Nav1.5 poses special quantitation challenges. To date, only the relative quantities of this protein have been measured in biological samples. Here, we describe the first targeted and mass spectrometry (MS)-based quantitative assay that can provide the copy numbers of Nav1.5 in cells with a well-defined lower limit of quantification (LLOQ) and precision. Applying the developed assay, we successfully quantified transiently expressed Nav1.5 in as few as 1.5 million Chinese hamster ovary (CHO) cells. The obtained quantity was 3 ± 2 fmol on the column and 3 ± 2 × 104 copies/cell. To our knowledge, this is the first absolute quantity of Nav1.5 measured in a biological sample.
Inhibition of RhoA and Cdc42 by miR-133a Modulates Retinoic Acid Signalling during Early Development of Posterior Cardiac Tube Segment
Authors: Garcia-Padilla Carlos , Garcia-Lopez Virginio , Aranega Amelia , Franco Diego , Garcia-Martinez Virginio , Lopez-Sanchez Carmen
Published: April 2022

It is well known that multiple microRNAs play crucial roles in cardiovascular development, including miR-133a. Additionally, retinoic acid regulates atrial marker expression. In order to analyse the role of miR-133a as a modulator of retinoic acid signalling during the posterior segment of heart tube formation, we performed functional experiments with miR-133a and retinoic acid by means of microinjections into the posterior cardiac precursors of both primitive endocardial tubes in chick embryos. Subsequently, we subjected embryos to whole mount in situ hybridisation, immunohistochemistry and qPCR analysis. Our results demonstrate that miR-133a represses RhoA and Cdc42, as well as Raldh2/Aldh1a2, and the specific atrial markers Tbx5 and AMHC1, which play a key role during differentiation. Furthermore, we observed that miR-133a upregulates p21 and downregulates cyclin A by repressing RhoA and Cdc42, respectively, thus functioning as a cell proliferation inhibitor. Additionally, retinoic acid represses miR-133a, while it increases Raldh2, Tbx5 and AMHC1. Given that RhoA and Cdc42 are involved in Raldh2 expression and that they are modulated by miR-133a, which is influenced by retinoic acid signalling, our results suggest the presence of a negative feedback mechanism between miR-133a and retinoic acid during early development of the posterior cardiac tube segment. Despite additional unexplored factors being possible contributors to this negative feedback mechanism, miR-133a might also be considered as a potential therapeutic tool for the diagnosis, therapy and prognosis of cardiac diseases.
Screening of Relevant Metabolism-Disrupting Chemicals on Pancreatic β-Cells: Evaluation of Murine and Human In Vitro Models
Authors: Al-Abdulla Ruba , Ferrero Hilda , Soriano Sergi , Boronat-Belda Talía , Alonso-Magdalena Paloma
Published: April 2022

Endocrine-disrupting chemicals (EDCs) are chemical substances that can interfere with the normal function of the endocrine system. EDCs are ubiquitous and can be found in a variety of consumer products such as food packaging materials, personal care and household products, plastic additives, and flame retardants. Over the last decade, the impact of EDCs on human health has been widely acknowledged as they have been associated with different endocrine diseases. Among them, a subset called metabolism-disrupting chemicals (MDCs) is able to promote metabolic changes that can lead to the development of metabolic disorders such as diabetes, obesity, hepatic steatosis, and metabolic syndrome, among others. Despite this, today, there are still no definitive and standardized in vitro tools to support the metabolic risk assessment of existing and emerging MDCs for regulatory purposes. Here, we evaluated the following two different pancreatic cell-based in vitro systems: the murine pancreatic β-cell line MIN6 as well as the human pancreatic β-cell line EndoC-βH1. Both were challenged with the following range of relevant concentrations of seven well-known EDCs: (bisphenol-A (BPA), bisphenol-S (BPS), bisphenol-F (BPF), perfluorooctanesulfonic acid (PFOS), di(2-ethylhexyl) phthalate (DEHP), cadmium chloride (CdCl2), and dichlorodiphenyldichloroethylene (DDE)). The screening revealed that most of the tested chemicals have detectable, deleterious effects on glucose-stimulated insulin release, insulin content, electrical activity, gene expression, and/or viability. Our data provide new molecular information on the direct effects of the selected chemicals on key aspects of pancreatic β-cell function, such as the stimulus-secretion coupling and ion channel activity. In addition, we found that, in general, the sensitivity and responses were comparable to those from other in vivo studies reported in the literature. Overall, our results suggest that both systems can serve as effective tools for the rapid screening of potential MDC effects on pancreatic β-cell physiology as well as for deciphering and better understanding the molecular mechanisms that underlie their action.
Drosophila as a Model System for Studying of the Evolution and Functional Specialization of the Y Chromosome
Authors: Kotov Alexei A. , Bazylev Sergei S. , Adashev Vladimir E. , Shatskikh Aleksei S. , Olenina Ludmila V.
Published: April 2022

The Y chromosome is one of the sex chromosomes found in males of animals of different taxa, including insects and mammals. Among all chromosomes, the Y chromosome is characterized by a unique chromatin landscape undergoing dynamic evolutionary change. Being entirely heterochromatic, the Y chromosome as a rule preserves few functional genes, but is enriched in tandem repeats and transposons. Due to difficulties in the assembly of the highly repetitive Y chromosome sequence, deep analyses of Y chromosome evolution, structure, and functions are limited to a few species, one of them being Drosophila melanogaster. Despite Y chromosomes exhibiting high structural divergence between even closely related species, Y-linked genes have evolved convergently and are mainly associated with spermatogenesis-related activities. This indicates that male-specific selection is a dominant force shaping evolution of Y chromosomes across species. This review presents our analysis of current knowledge concerning Y chromosome functions, focusing on recent findings in Drosophila. Here we dissect the experimental and bioinformatics data about the Y chromosome accumulated to date in Drosophila species, providing comparative analysis with mammals, and discussing the relevance of our analysis to a wide range of eukaryotic organisms, including humans.
Metal Coordination Enhances Chalcogen Bonds: CSD Survey and Theoretical Calculations
Authors: Frontera Antonio , Bauza Antonio
Published: April 2022

In this study the ability of metal coordinated Chalcogen (Ch) atoms to undergo Chalcogen bonding (ChB) interactions has been evaluated at the PBE0-D3/def2-TZVP level of theory. An initial CSD (Cambridge Structural Database) inspection revealed the presence of square planar Pd/Pt coordination complexes where divalent Ch atoms (Se/Te) were used as ligands. Interestingly, the coordination to the metal center enhanced the σ-hole donor ability of the Ch atom, which participates in ChBs with neighboring units present in the X-ray crystal structure, therefore dictating the solid state architecture. The X-ray analyses were complemented with a computational study (PBE0-D3/def2-TZVP level of theory), which shed light into the strength and directionality of the ChBs studied herein. Owing to the new possibilities that metal coordination offers to enhance or modulate the σ-hole donor ability of Chs, we believe that the findings presented herein are of remarkable importance for supramolecular chemists as well as for those scientists working in the field of solid state chemistry.
Autophagy Signaling by Neural-Induced Human Adipose Tissue-Derived Stem Cell-Conditioned Medium during Rotenone-Induced Toxicity in SH-SY5Y Cells
Authors: Ramalingam Mahesh , Jeong Han-Seong , Hwang Jinsu , Cho Hyong-Ho , Kim Byeong C. , Kim Eungpil , Jang Sujeong
Published: April 2022

Rotenone (ROT) inhibits mitochondrial complex I, leading to reactive oxygen species formation, which causes neurodegeneration and alpha-synuclein (α-syn) aggregation and, consequently, Parkinson’s disease. We previously found that a neurogenic differentiated human adipose tissue-derived stem cell-conditioned medium (NI-hADSC-CM) was protective against ROT-induced toxicity in SH-SY5Y cells. In the present study, ROT significantly decreased the phospho (p)-mTORC1/total (t)-mTOR, p-mTORC2/t-mTOR, and p-/t-ULK1 ratios and the ATG13 level by increasing the DEPTOR level and p-/t-AMPK ratio. Moreover, ROT increased the p-/t-Akt ratio and glycogen synthase kinase-3β (GSK3β) activity by decreasing the p-/t-ERK1/2 ratios and beclin-1 level. ROT also promoted the lipidation of LC3B-I to LC3B-II by inducing autophagosome formation in Triton X-100-soluble and -insoluble cell lysate fractions. Additionally, the levels of ATG3, 5, 7, and 12 were decreased, along with those of lysosomal LAMP1, LAMP2, and TFEB, leading to lysosomal dysfunction. However, NI-hADSC-CM treatment increased the p-mTORC1, p-mTORC2, p-ULK1, p-Akt, p-ERK1/2, ATG13, and beclin-1 levels and decreased the p-AMPK level and GSK3β activity in response to ROT-induced toxicity. Additionally, NI-hADSC-CM restored the LC3B-I level, increased the p62 level, and normalized the ATG and lysosomal protein amounts to control levels. Autophagy array revealed that the secreted proteins in NI-hADSC-CM could be crucial in the neuroprotection. Taken together, our results showed that the neuroprotective effects of NI-hADSC-CM on the autophagy signaling pathways could alleviate the aggregation of α-syn in Parkinson’s disease and other neurodegenerative disorders.
Neural Network Aided Detection of Huntington Disease
Authors: Alfonso Perez Gerardo , Caballero Villarraso Javier
Published: April 2022

Huntington Disease (HD) is a degenerative neurological disease that causes a significant impact on the quality of life of the patient and eventually death. In this paper we present an approach to create a biomarker using as an input DNA CpG methylation data to identify HD patients. DNA CpG methylation is a well-known epigenetic marker for disease state. Technological advances have made it possible to quickly analyze hundreds of thousands of CpGs. This large amount of information might introduce noise as potentially not all DNA CpG methylation levels will be related to the presence of the illness. In this paper, we were able to reduce the number of CpGs considered from hundreds of thousands to 237 using a non-linear approach. It will be shown that using only these 237 CpGs and non-linear techniques such as artificial neural networks makes it possible to accurately differentiate between control and HD patients. An underlying assumption in this paper is that there are no indications suggesting that the process is linear and therefore non-linear techniques, such as artificial neural networks, are a valid tool to analyze this complex disease. The proposed approach is able to accurately distinguish between control and HD patients using DNA CpG methylation data as an input and non-linear forecasting techniques. It should be noted that the dataset analyzed is relatively small. However, the results seem relatively consistent and the analysis can be repeated with larger data-sets as they become available.
Beneficial Effects of Adjusted Perfusion and Defibrillation Strategies on Rhythm Control within Controlled Automated Reperfusion of the Whole Body (CARL) for Refractory Out-of-Hospital Cardiac Arrest
Authors: Brixius Sam Joé , Pooth Jan-Steffen , Haberstroh Jörg , Damjanovic Domagoj , Scherer Christian , Greiner Philipp , Benk Christoph , Beyersdorf Friedhelm , Trummer Georg
Published: April 2022

Survival and neurological outcomes after out-of-hospital cardiac arrest (OHCA) remain low. The further development of prehospital extracorporeal resuscitation (ECPR) towards Controlled Automated Reperfusion of the Whole Body (CARL) has the potential to improve survival and outcome in these patients. In CARL therapy, pulsatile, high blood-flow reperfusion is performed combined with several modified reperfusion parameters and adjusted defibrillation strategies. We aimed to investigate whether pulsatile, high-flow reperfusion is feasible in refractory OHCA and whether the CARL approach improves heart-rhythm control during ECPR. In a reality-based porcine model of refractory OHCA, 20 pigs underwent prehospital CARL or conventional ECPR. Significantly higher pulsatile blood-flow proved to be feasible, and critical hypotension was consistently prevented via CARL. In the CARL group, spontaneous rhythm conversions were observed using a modified priming solution. Applying potassium-induced secondary cardioplegia proved to be a safe and effective method for sustained rhythm conversion. Moreover, significantly fewer defibrillation attempts were needed, and cardiac arrhythmias were reduced during reperfusion via CARL. Prehospital CARL therapy thus not only proved to be feasible after prolonged OHCA, but it turned out to be superior to conventional ECPR regarding rhythm control.
Identification of Filamentous Fungi by MALDI-TOF Mass Spectrometry: Evaluation of Three Different Sample Preparation Methods and Validation of an In-House Species Cutoff
Authors: Honsig Claudia , Selitsch Brigitte , Hollenstein Marlene , Vossen Matthias G. , Spettel Kathrin , Willinger Birgit
Published: April 2022

Invasive infections caused by filamentous fungi constitute a leading cause of morbidity and mortality in immunocompromised patients. Rapid and reliable identification of filamentous fungi is essential for the early initiation of appropriate treatment. In the present study, 230 filamentous fungi isolates identified by conventional methods were investigated using MALDI-TOF MS (Bruker Daltonics, Bremen, Germany) in combination with the Filamentous Fungi Library 3.0 provided by the manufacturer. Three different sample preparation methods were applied as recommended by the manufacturer and identification rates were compared using the criteria provided by the manufacturer. Application of the more time-consuming sample preparation methods clearly improved identification at the species level. Thus, the identification rate increased from 48.9% using the simplest method to 76.1% with the most laborious procedure. Misidentifications did not occur. Furthermore, the reliability of an in-house threshold for species identification was investigated. The reduced threshold increased the rate of isolates correctly identified at the species level by up to 86.4%. As no misidentification was made at the genus level and only one misidentification of minor significance occurred at the species level, this threshold could be validated for routine use in our laboratory. In conclusion, regarding the high identification rates achieved, this commercial platform proved suitable for implementation in routine diagnosis.
Systemic Infection by Non-albicans Candida Species Affects the Development of a Murine Model of Multiple Sclerosis
Authors: Fraga-Silva Thais Fernanda de Campos , Munhoz-Alves Natália , Mimura Luiza Ayumi Nishiyama , Oliveira Larissa Ragozo Cardoso de , Figueiredo-Godoi Lívia Mara Alves , Garcia Maíra Terra , Oliveira Evelyn Silva , Ishikawa Larissa Lumi Watanabe , Zorzella-Pezavento Sofia Fernanda Gonçalves , Bonato Vânia Luiza Deperon , Junqueira Juliana Campos , Bagagli Eduardo , Sartori Alexandrina
Published: April 2022

Candidiasis may affect the central nervous system (CNS), and although Candida albicans is predominant, non-albicans Candida species can also be associated with CNS infections. Some studies have suggested that Candida infections could increase the odds of multiple sclerosis (MS) development. In this context, we investigated whether systemic infection by non-albicans Candida species would affect, clinically or immunologically, the severity of experimental autoimmune encephalomyelitis (EAE), which is an animal model used to study MS. For this, a strain of C. glabrata, C. krusei, and C. parapsilosis was selected and characterized using different in vitro and in vivo models. In these analysis, all the strains exhibited the ability to form biofilms, produce proteolytic enzymes, and cause systemic infections in Galleria mellonella, with C. glabrata being the most virulent species. Next, C57BL/6 mice were infected with strains of C. glabrata, C. krusei, or C. parapsilosis, and 3 days later were immunized with myelin oligodendrocyte glycoprotein to develop EAE. Mice from EAE groups previously infected with C. glabrata and C. krusei developed more severe and more prevalent paralysis, while mice from the EAE group infected with C. parapsilosis developed a disease comparable to non-infected EAE mice. Disease aggravation by C. glabrata and C. krusei strains was concomitant to increased IL-17 and IFN-γ production by splenic cells stimulated with fungi-derived antigens and with increased percentage of T lymphocytes and myeloid cells in the CNS. Analysis of interaction with BV-2 microglial cell line also revealed differences among these strains, in which C. krusei was the strongest activator of microglia concerning the expression of MHC II and CD40 and pro-inflammatory cytokine production. Altogether, these results indicated that the three non-albicans Candida strains were similarly able to reach the CNS but distinct in terms of their effect over EAE development. Whereas C. glabrata and C. Krusei aggravated the development of EAE, C. parapsilosis did not affect its severity. Disease worsening was partially associated to virulence factors in C. glabrata and to a strong activation of microglia in C. krusei infection. In conclusion, systemic infections by non-albicans Candida strains exerted influence on the experimental autoimmune encephalomyelitis in both immunological and clinical aspects, emphasizing their possible relevance in MS development.