bluuebunny/crossref_metadata_2025 huggingface dataset:
Risk Perceptions of Low Nicotine Cigarettes and Alternative Nicotine Products across Priority Smoking Populations
Authors: Denlinger-Apte Rachel L. , Pacek Lauren R. , Ross Jennifer Cornacchione , Bansal-Travers Maansi , Donny Eric C. , Hatsukami Dorothy K. , Carroll Dana Mowls
Published: May 2021

Background: As the U.S. Food and Drug Administration considers a low nicotine product standard for cigarettes, it is important to examine how people who smoke, especially individuals from priority populations disproportionately affected by smoking, perceive low nicotine content (LNC) cigarettes and their relative risk perceptions of alternative nicotine delivery system (ANDS) products, including e-cigarettes and snus, and medicinal nicotine. Methods: Data are from Wave 4 (2016–2017) of the adult Population Assessment of Tobacco Use and Health (PATH) Study. We examined respondents’ absolute risk perceptions about nicotine, LNC cigarettes, ANDS products and medicinal nicotine; their relative risk perceptions of LNC cigarettes and ANDS products compared to conventional cigarettes; and their relative risk perceptions of medicinal nicotine compared to ANDS products. Results: The majority of respondents across priority smoking populations indicated snus, e-cigarettes, and LNC cigarettes were ‘about the same’ level of harmfulness or addictiveness as conventional cigarettes. The majority of respondents indicated e-cigarettes to be ‘about the same’ harmfulness as medicinal nicotine. Conclusions: Our study indicates that adults who smoke cigarettes generally have misperceptions about the harms of nicotine and the relative risks of ANDS products and such misperceptions exist regardless of their racial/ethnic identity, sexual orientation, and gender identity.
Functional Food—Consumer Motivations and Expectations
Authors: Topolska Kinga , Florkiewicz Adam , Filipiak-Florkiewicz Agnieszka
Published: May 2021

This review provides insight into consumer attitudes toward functional food (FF), with the purpose of better understanding the needs and behavior regarding this kind of product. A total of 47 articles were selected for this paper. The available studies from last 20 years differ according to the focus (awareness, attitudes, motivations, willingness, acceptance by consumers) and methodologies used. Several factors, including socio-demographic, cognitive and attitudinal ones, seem to be serve as the basis for the acceptance of functional products. The research papers showed that nutritional knowledge is the most important of these. Older people are more interested in functional products than younger consumers, because of their stronger belief in the health benefits of FF. Moreover, women are more open to compromise between taste and health properties. Claims concerning the disease preventative properties of FF are the most attractive for consumers. This review focuses also on future perspectives for the functional food market. Adequate knowledge and evidence-based communication seem to be the most promising ways to increase consumers’ interest in these kinds of products.
Risk Prediction of Barrett’s Esophagus in a Taiwanese Health Examination Center Based on Regression Models
Authors: Lin Po-Hsiang , Hsieh Jer-Guang , Yu Hsien-Chung , Jeng Jyh-Horng , Hsu Chiao-Lin , Chen Chien-Hua , Wu Pin-Chieh
Published: May 2021

Determining the target population for the screening of Barrett’s esophagus (BE), a precancerous condition of esophageal adenocarcinoma, remains a challenge in Asia. The aim of our study was to develop risk prediction models for BE using logistic regression (LR) and artificial neural network (ANN) methods. Their predictive performances were compared. We retrospectively analyzed 9646 adults aged ≥20 years undergoing upper gastrointestinal endoscopy at a health examinations center in Taiwan. Evaluated by using 10-fold cross-validation, both models exhibited good discriminative power, with comparable area under curve (AUC) for the LR and ANN models (Both AUC were 0.702). Our risk prediction models for BE were developed from individuals with or without clinical indications of upper gastrointestinal endoscopy. The models have the potential to serve as a practical tool for identifying high-risk individuals of BE among the general population for endoscopic screening.
Missing Diagnoses during the COVID-19 Pandemic: A Year in Review
Authors: Pifarré i Arolas Héctor , Vidal-Alaball Josep , Gil Joan , López Francesc , Nicodemo Catia , Saez Marc
Published: May 2021

The COVID-19 pandemic has had major impacts on population health not only through COVID-positive cases, but also via the disruption of healthcare services, which in turn has impacted the diagnosis and treatment of all other diseases during this time. We study changes in all new registered diagnoses in ICD-10 groups during 2020 with respect to a 2019 baseline. We compare new diagnoses in 2019 and 2020 based on administrative records of the public primary health system in Central Catalonia, Spain, which cover over 400,000 patients and 3 million patient visits. We study the ratio of new diagnoses between 2019 and 2020 and find an average decline of 31.1% in new diagnoses, with substantial drops in April (61.1%), May (55.6%), and November (52%). Neoplasms experience the largest decline (49.7%), with heterogeneity in the magnitudes of the declines across different types of cancer diagnoses. While we find evidence of temporal variation in new diagnoses, reductions in diagnoses early in the year are not recouped by the year end. The observed decline in new diagnoses across all diagnosis groups suggest a large number of untreated and undetected cases across conditions. Our findings provide a year-end summary of the impact of the pandemic on healthcare activities and can help guide health authorities to design evidence-based plans to target under-diagnosed conditions in 2021.
Factors Associated with Regular Zumba Practice as Preliminary Results: A Population-Based Approach in Cebu Province, the Philippines
Authors: Yamasaki Junko , Sakisaka Kayako , Mission Parolita A. , Soluta Nasudi G. , delos Santos Norre Jean V. , Palaca Julie Vannie , Pacalioga Retz Pol O.
Published: May 2021

The prevalence of overweight/obesity in the adult population in the Philippines has doubled in the past 20 years. Zumba exercise has recently been implemented throughout the Philippines. However, there is scarce information on the effects of Zumba on obesity and Zumba participants’ characteristics in the Philippines. This study described the current practice of Zumba in the Philippines, along with the practitioners’ characteristics, and identified factors associated with Zumba participation. In this observational, cross-sectional study, a structured questionnaire was used to survey 10 Zumba locations in September 2019. Anthropometric measurements of participants were assessed. Respondents included 171 women (88.6%) and 22 men (11.4%), with a mean (±standard deviation [SD]) age of 44.1 (±8.9) years. All respondents answered that Zumba was enjoyable, and some answered “very enjoyable”. Determinants of frequent participation were as follows: being older than the mean age of participants, starting Zumba to enjoy dancing, starting Zumba not to lose weight, shopping mall location, and participation fee required. “To enjoy dance” being a motivation for Zumba practice was identified as a determinant of frequent participation rather than “to lose weight.” The element of “enjoyable” may strongly influence the continuation and frequent participation of Zumba exercise in the Philippines.
Disparities in HIV Clinical Stages Progression of Patients at Outpatient Clinics in Democratic Republic of Congo
Authors: Ewetola Raimi , Shah Gulzar H. , Maluantesa Lievain , Etheredge Gina , Waterfield Kristie , Mulenga Astrid , Kilundu Apolinaire
Published: May 2021

Context: In this era of patient-centered care, it is increasingly important for HIV/AIDS care and treatment programs to customize their services according to patients’ clinical stage progression and other risk assessments. To enable such customization of HIV care and treatment delivery, the research evidence explaining factors associated with patients’ clinical stages is needed. Objectives: The primary objective of this study was to produce such scientific evidence by analyzing the most recent data for patients at outpatient clinics in the provinces of Kinshasa and Haut-Katanga and to examine the patient characteristics associated with WHO stages of disease progression. Methods: Using a quantitative retrospective cohort study design, we analyzed data from 49,460 people living with HIV (PLHIV) on antiretroviral therapy (ART) from 241 HIV/AIDS clinics located in Haut-Katanga and Kinshasa provinces of the Democratic Republic of Congo. We performed Chi-square and multinomial logistic regression analyses. Results: A small proportion (i.e., 4.4%) of PLHIV were at WHO’s clinical progression stage 4, whereas 30.7% were at clinical stage 3, another 22.9% at stage 2, and the remaining 41.9% were at stage 1, the least severe stage. After controlling for other demographic and clinical factors included in the model, the likelihood of being at stage 1 rather than stage 3 or 4 was significantly higher (at p ≤ 0.05) for patients with no tuberculosis (TB) than those with TB co-infection (adjusted odds ratio or AOR, 5.73; confidence interval or CI, 4.98–6.59). The odds of being at stage 1 were significantly higher for female patients (AOR, 1.35; CI, 1.29–1.42), and those with the shorter duration on ART (vs. greater than 40.37 months). Patents in rural health zones (AOR, 0.32) and semi-rural health zones (AOR, 0.79) were less likely to be at stage 1, compared to patients in urban health zones. Conclusions: Our study showed that TB co-infection raised the risk for PLHIV to be at the severe stages of clinical progression of HIV. Such variation supports the thesis that customized HIV management approaches and clinical regimens may be imperative for this high-risk population. We also found significant variation in HIV clinical progression stages by geographic location and demographic characteristics. Such variation points to the need for more targeted efforts to address the disparities, as the programs attempt to improve the effectiveness of HIV care and treatment. The intersectionality of vulnerabilities from HIV, TB, and COVID-19-related hardships has elevated the need for customized care and treatment even more in the COVID-19 era.
Effects of Soy Isoflavones on Biochemical Markers of Bone Metabolism in Postmenopausal Women: A Systematic Review and Meta-Analysis of Randomized Controlled Trials
Authors: Kanadys Wiesław , Barańska Agnieszka , Błaszczuk Agata , Polz-Dacewicz Małgorzata , Drop Bartłomiej , Malm Maria , Kanecki Krzysztof
Published: May 2021

This systematic review and meta-analysis of randomized controlled trials was performed to more completely assess potential changes in bone turnover marker levels in postmenopausal women during the intake of soy isoflavones. PubMed (Medline) and EMBASE were searched for relevant studies, and their quality was evaluated according to Cochrane criteria. The levels of markers were evaluated in a total of 1114 women who ingested mean daily doses of 98.2 mg (30.9 to 300) of soy isoflavones for 3 to 24 months, in comparison to those of 1081 subjects who used a placebo. Ten, eighteen, eight, and fourteen comparison studies were finally selected for an estimation of the effects on osteocalcin (OC), bone alkaline phosphatase (BAP), pyridinoline (PYD), and deoxypyridinoline (DPD), respectively. A summary of the results of intervention was as follows: 4.16%, 95% CI: –7.72–16.04, p = 0.49 for OC; 5.50%, 95% CI: –3.81–14.82, p = 0.25 for BAP; –12.09%, 95% CI: –25.37–1.20, p = 0.07 for PYD; and –7.48%, 95% CI: –15.37–0.41, p = 0.06 for DPD. The meta-analysis of the included studies revealed some statistically insignificant observations that soy isoflavones intake is associated with a trend in increased levels of OC and BAP, as well as a trend in reduced levels of PYD and DPD. Soy isoflavones may have a beneficial effect on bone formation markers, but this requires extensive multi-center research.
Palliative Care Professionals’ Message to Others: An Ethnographic Approach
Authors: Reigada Carla , Centeno Carlos , Gonçalves Edna , Arantzamendi Maria
Published: May 2021

Introduction: Palliative care continues to be misunderstood within the world of healthcare. Palliative care professionals are key agents for promoting a greater understanding of their field. This study aims to examine the messages, both implicit and explicit, that palliative care professionals transmit about themselves and their work within their teams and to other health professionals. Methods: Focused ethnographic secondary analysis, exploring the interactions of palliative care professionals as it happens at everyday work. An inductive thematic analysis was developed from 242 h of observation of the daily work practices of palliative care professionals, focusing on their interactions with others. The data was coded without predefined categories, and the analysis was performed independently by two researchers. Results: Palliative professionals communicate that they are part of an active team working in an organized manner. They value and feel proud of their work. Despite the intensity of their work, these professionals are always available to others, to whom they demonstrate a clear professional identity. They convey their expertise in alleviating suffering, respectful behavior and collaborative ability. Conclusion: Professionals, in their daily work, communicate through their messages the essence of palliative care. It is essential that palliative care professionals perceive themselves as potential influencers and explicitly transmit the reasons for their intervention. Otherwise, others will perpetuate the myths, misunderstandings, and lack of a positive reputation for palliative care.
A Dynamic and Static Context-Aware Attention Network for Trajectory Prediction
Authors: Yu Jian , Zhou Meng , Wang Xin , Pu Guoliang , Cheng Chengqi , Chen Bo
Published: May 2021

Forecasting the motion of surrounding vehicles is necessary for an autonomous driving system applied in complex traffic. Trajectory prediction helps vehicles make more sensible decisions, which provides vehicles with foresight. However, traditional models consider the trajectory prediction as a simple sequence prediction task. The ignorance of inter-vehicle interaction and environment influence degrades these models in real-world datasets. To address this issue, we propose a novel Dynamic and Static Context-aware Attention Network named DSCAN in this paper. The DSCAN utilizes an attention mechanism to dynamically decide which surrounding vehicles are more important at the moment. We also equip the DSCAN with a constraint network to consider the static environment information. We conducted a series of experiments on a real-world dataset, and the experimental results demonstrated the effectiveness of our model. Moreover, the present study suggests that the attention mechanism and static constraints enhance the prediction results.
Subsurface Topographic Modeling Using Geospatial and Data Driven Algorithm
Authors: Abbaszadeh Shahri Abbas , Kheiri Ali , Hamzeh Aliakbar
Published: May 2021

Infrastructures play an important role in urbanization and economic activities but are vulnerable. Due to unavailability of accurate subsurface infrastructure maps, ensuring the sustainability and resilience often are poorly recognized. In the current paper a 3D topographical predictive model using distributed geospatial data incorporated with evolutionary gene expression programming (GEP) was developed and applied on a concrete-face rockfill dam (CFRD) in Guilan province- northern to generate spatial variation of the subsurface bedrock topography. The compared proficiency of the GEP model with geostatistical ordinary kriging (OK) using different analytical indexes showed 82.53% accuracy performance and 9.61% improvement in precisely labeled data. The achievements imply that the retrieved GEP model efficiently can provide accurate enough prediction and consequently meliorate the visualization insights linking the natural and engineering concerns. Accordingly, the generated subsurface bedrock model dedicates great information on stability of structures and hydrogeological properties, thus adopting appropriate foundations.
The Land-Use Change Dynamics Based on the CORINE Data in the Period 1990–2018 in the European Archipelagos of the Macaronesia Region: Azores, Canary Islands, and Madeira
Authors: Castanho Rui Alexandre , Naranjo Gomez José Manuel , Vulevic Ana , Couto Gualter
Published: May 2021

Islands as peripheral and ultra-peripheral are typically highlighted as ecologically sensitive areas to human activities due to the tremendous biological diversity of beings and the future possibility of habitat loss. In this regard, the comprehension of the land occupation dynamics and trends in the ultra-peripheral territories is crucial to attempt long-lasting regional sustainability, as is the island region’s case. Therefore, the present article aims to analyze the trends and dynamics of the land-use changes on the European Archipelagos of the Macaronesia Region over the last three decades, using the CORINE (Coordination of Information on the Environment) data. Some of the obtained results show that about 3.4% of the Azores’ surface is characterized mainly by discontinuous urban fabric, representing 67% of the total urban fabric of the Azores over the last thirty years. Additionally, in Madeira Archipelago, the land is mainly occupied by forest and semi-natural areas, representing almost three-thirds of the territory. A similar scenario is verified in the Canary Islands, where forests and semi-natural areas represent approximately three-quarters of the territory. Once more, this study shows the relevance of the island areas’ unique character, which should be preserved and protected. Therefore, the priorities must be defined and established management strategies that are significant for the well-being of these highly valued areas. Moreover, the study showed that notable changes had occurred in the period 1990–2018 in this landscape. Hence there is a need for appropriate measures to mitigate these negative impacts on the environment.
Double p52Shc/p46Shc Rat Knockout Demonstrates Severe Gait Abnormalities Accompanied by Dilated Cardiomyopathy
Authors: Miller Bradley , Kostrominova Tatiana Y. , Geurts Aron M. , Sorokin Andrey
Published: May 2021

The ubiquitously expressed adaptor protein Shc exists in three isoforms p46Shc, p52Shc, and p66Shc, which execute distinctly different actions in cells. The role of p46Shc is insufficiently studied, and the purpose of this study was to further investigate its functional significance. We developed unique rat mutants lacking p52Shc and p46Shc isoforms (p52Shc/46Shc-KO) and carried out histological analysis of skeletal and cardiac muscle of parental and genetically modified rats with impaired gait. p52Shc/46Shc-KO rats demonstrate severe functional abnormalities associated with impaired gait. Our analysis of p52Shc/46Shc-KO rat axons and myelin sheets in cross-sections of the sciatic nerve revealed the presence of significant anomalies. Based on the lack of skeletal muscle fiber atrophy and the presence of sciatic nerve abnormalities, we suggest that the impaired gait in p52Shc/46Shc-KO rats might be due to the sensory feedback from active muscle to the brain locomotor centers. The lack of dystrophin in some heart muscle fibers reflects damage due to dilated cardiomyopathy. Since rats with only p52Shc knockout do not display the phenotype of p52Shc/p46Shc-KO, abnormal locomotion is likely to be caused by p46Shc deletion. Our data suggest a previously unknown role of 46Shc actions and signaling in regulation of gait.
Adenovirus Structure: What Is New?
Authors: Gallardo José , Pérez-Illana Marta , Martín-González Natalia , San Martín Carmen
Published: May 2021

Adenoviruses are large (~950 Å) and complex non-enveloped, dsDNA icosahedral viruses. They have a pseudo-T = 25 triangulation number with at least 12 different proteins composing the virion. These include the major and minor capsid proteins, core proteins, maturation protease, terminal protein, and packaging machinery. Although adenoviruses have been studied for more than 60 years, deciphering their architecture has presented a challenge for structural biology techniques. An outstanding event was the first near-atomic resolution structure of human adenovirus type 5 (HAdV-C5), solved by cryo-electron microscopy (cryo-EM) in 2010. Discovery of new adenovirus types, together with methodological advances in structural biology techniques, in particular cryo-EM, has lately produced a considerable amount of new, high-resolution data on the organization of adenoviruses belonging to different species. In spite of these advances, the organization of the non-icosahedral core is still a great unknown. Nevertheless, alternative techniques such as atomic force microscopy (AFM) are providing interesting glimpses on the role of the core proteins in genome condensation and virion stability. Here we summarize the current knowledge on adenovirus structure, with an emphasis on high-resolution structures obtained since 2010.
Do Autism Spectrum and Autoimmune Disorders Share Predisposition Gene Signature Due to mTOR Signaling Pathway Controlling Expression?
Authors: Trifonova Ekaterina A. , Klimenko Alexandra I. , Mustafin Zakhar S. , Lashin Sergey A. , Kochetov Alex V.
Published: May 2021

Autism spectrum disorder (ASD) is characterized by uncommon genetic heterogeneity and a high heritability concurrently. Most autoimmune disorders (AID), similarly to ASD, are characterized by impressive genetic heterogeneity and heritability. We conducted gene-set analyses and revealed that 584 out of 992 genes (59%) included in a new release of the SFARI Gene database and 439 out of 871 AID-associated genes (50%) could be attributed to one of four groups: 1. FMRP (fragile X mental retardation protein) target genes, 2. mTOR signaling network genes, 3. mTOR-modulated genes, and 4. vitamin D3-sensitive genes. With the exception of FMRP targets, which are obviously associated with the direct involvement of local translation disturbance in the pathological mechanisms of ASD, the remaining categories are represented among AID genes in a very similar percentage as among ASD predisposition genes. Thus, mTOR signaling pathway genes make up 4% of ASD and 3% of AID genes, mTOR-modulated genes—31% of both ASD and AID genes, and vitamin D-sensitive genes—20% of ASD and 23% of AID genes. The network analysis revealed 3124 interactions between 528 out of 729 AID genes for the 0.7 cutoff, so the great majority (up to 67%) of AID genes are related to the mTOR signaling pathway directly or indirectly. Our present research and available published data allow us to hypothesize that both a certain part of ASD and AID comprise a connected set of disorders sharing a common aberrant pathway (mTOR signaling) rather than a vast set of different disorders. Furthermore, an immune subtype of the autism spectrum might be a specific type of autoimmune disorder with an early manifestation of a unique set of predominantly behavioral symptoms.
NO● Represses the Oxygenation of Arachidonoyl PE by 15LOX/PEBP1: Mechanism and Role in Ferroptosis
Authors: Mikulska-Ruminska Karolina , Anthonymuthu Tamil , Levkina Anastasia , Shrivastava Indira , Kapralov Alexandr , Bayır Hülya , Kagan Valerian , Bahar Ivet
Published: May 2021

We recently discovered an anti-ferroptotic mechanism inherent to M1 macrophages whereby high levels of NO● suppressed ferroptosis via inhibition of hydroperoxy-eicosatetraenoyl-phosphatidylethanolamine (HpETE-PE) production by 15-lipoxygenase (15LOX) complexed with PE-binding protein 1 (PEBP1). However, the mechanism of NO● interference with 15LOX/PEBP1 activity remained unclear. Here, we use a biochemical model of recombinant 15LOX-2 complexed with PEBP1, LC-MS redox lipidomics, and structure-based modeling and simulations to uncover the mechanism through which NO● suppresses ETE-PE oxidation. Our study reveals that O2 and NO● use the same entry pores and channels connecting to 15LOX-2 catalytic site, resulting in a competition for the catalytic site. We identified residues that direct O2 and NO● to the catalytic site, as well as those stabilizing the esterified ETE-PE phospholipid tail. The functional significance of these residues is supported by in silico saturation mutagenesis. We detected nitrosylated PE species in a biochemical system consisting of 15LOX-2/PEBP1 and NO● donor and in RAW264.7 M2 macrophages treated with ferroptosis-inducer RSL3 in the presence of NO●, in further support of the ability of NO● to diffuse to, and react at, the 15LOX-2 catalytic site. The results provide first insights into the molecular mechanism of repression of the ferroptotic Hp-ETE-PE production by NO●.
PSII Activity Was Inhibited at Flowering Stage with Developing Black Bracts of Oat
Authors: Liu Bei , Zhang Di , Sun Ming , Li Manli , Ma Xiqing , Jia Shangang , Mao Peisheng
Published: May 2021

The color of bracts generally turns yellow or black from green during cereal grain development. However, the impact of these phenotypic changes on photosynthetic physiology during black bract formation remains unclear. Two oat cultivars (Avena sativa L.), ‘Triple Crown’ and ‘Qinghai 444’, with yellow and black bracts, respectively, were found to both have green bracts at the heading stage, but started to turn black at the flowering stage and become blackened at the milk stage for ‘Qinghai 444’. Their photosynthetic characteristics were analyzed and compared, and the key genes, proteins and regulatory pathways affecting photosynthetic physiology were determined in ‘Triple Crown’ and ‘Qinghai 444’ bracts. The results show that the actual PSII photochemical efficiency and PSII electron transfer rate of ‘Qinghai 444’ bracts had no significant changes at the heading and milk stages but decreased significantly (p < 0.05) at the flowering stage compared with ‘Triple Crown’. The chlorophyll content decreased, the LHCII involved in the assembly of supercomplexes in the thylakoid membrane was inhibited, and the expression of Lhcb1 and Lhcb5 was downregulated at the flowering stage. During this critical stage, the expression of Bh4 and C4H was upregulated, and the biosynthetic pathway of p-coumaric acid using tyrosine and phenylalanine as precursors was also enhanced. Moreover, the key upregulated genes (CHS, CHI and F3H) of anthocyanin biosynthesis might complement the impaired PSII activity until recovered at the milk stage. These findings provide a new insight into how photosynthesis alters during the process of oat bract color transition to black.
The Multifunctional Role of EMP3 in the Regulation of Membrane Receptors Associated with IDH-Wild-Type Glioblastoma
Authors: Martija Antoni Andreu , Pusch Stefan
Published: May 2021

Epithelial membrane protein 3 (EMP3) is a tetraspan membrane protein overexpressed in isocitrate dehydrogenase-wild-type (IDH-wt) glioblastoma (GBM). Several studies reported high EMP3 levels as a poor prognostic factor in GBM patients. Experimental findings based on glioma and non-glioma models have demonstrated the role of EMP3 in the regulation of several membrane proteins known to drive IDH-wt GBM. In this review, we summarize what is currently known about EMP3 biology. We discuss the regulatory effects that EMP3 exerts on a variety of oncogenic receptors and discuss how these mechanisms may relate to IDH-wt GBM. Lastly, we enumerate the open questions towards EMP3 function in IDH-wt GBM.
Soluble Prokaryotic Overexpression and Purification of Human GM-CSF Using the Protein Disulfide Isomerase b′a′ Domain
Authors: Nguyen Thi Kieu Oanh , Vu Thi Luong , Nguyen Minh Quan , Ta Huynh Kim Khanh , Park Kyoung Sun , Kim Soo Hyeon , Kim Chong Jai , Jang Yeon Jin , Choe Han
Published: May 2021

Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a member of the colony-stimulating factor (CSF) family, which functions to enhance the proliferation and differentiation of hematopoietic stem cells and other hematopoietic lineages such as neutrophils, dendritic cells, or macrophages. These proteins have thus generated considerable interest in clinical therapy research. A current obstacle to the prokaryotic production of human GM-CSF (hGM-CSF) is its low solubility when overexpressed and subsequent complex refolding processes. In our present study, the solubility of hGM-CSF was examined when combined with three N-terminal fusion tags in five E. coli strains at three different expression temperatures. In the five E. coli strains BL21 (DE3), ClearColi BL21 (DE3), LOBSTR, SHuffle T7 and Origami2 (DE3), the hexahistidine-tagged hGM-CSF showed the best expression but was insoluble in all cases at each examined temperature. Tagging with the maltose-binding protein (MBP) and the b′a′ domain of protein disulfide isomerase (PDIb′a′) greatly improved the soluble overexpression of hGM-CSF at 30 °C and 18 °C. The solubility was not improved using the Origami2 (DE3) and SHuffle T7 strains that have been engineered for disulfide bond formation. Two conventional chromatographic steps were used to purify hGM-CSF from the overexpressed PDIb′a′-hGM-CSF produced in ClearColi BL21 (DE3). In the experiment, 0.65 mg of hGM-CSF was isolated from a 0.5 L flask culture of these E. coli and showed a 98% purity by SDS-PAGE analysis and silver staining. The bioactivity of this purified hGM-CSF was measured at an EC50 of 16.4 ± 2 pM by a CCK8 assay in TF-1 human erythroleukemia cells.
ICAN (TRPM4) Contributes to the Intrinsic Excitability of Prefrontal Cortex Layer 2/3 Pyramidal Neurons
Authors: Riquelme Denise , Peralta Francisco , Navarro Franco , Moreno Claudio , Leiva-Salcedo Elias
Published: May 2021

Pyramidal neurons in the medial prefrontal cortical layer 2/3 are an essential contributor to the cellular basis of working memory; thus, changes in their intrinsic excitability critically affect medial prefrontal cortex (mPFC) functional properties. Transient Receptor Potential Melastatin 4 (TRPM4), a calcium-activated nonselective cation channel (CAN), regulates the membrane potential in a calcium-dependent manner. In this study, we uncovered the role of TRPM4 in regulating the intrinsic excitability plasticity of pyramidal neurons in the mouse mPFC layer of 2/3 using a combination of conventional and nystatin perforated whole-cell recordings. Interestingly, we found that TRPM4 is open at resting membrane potential, and its inhibition increases input resistance and hyperpolarizes membrane potential. After high-frequency stimulation, pyramidal neurons increase a calcium-activated non-selective cation current, increase the action potential firing, and the amplitude of the afterdepolarization, these effects depend on intracellular calcium. Furthermore, pharmacological inhibition or genetic silencing of TRPM4 reduces the firing rate and the afterdepolarization after high frequency stimulation. Together, these results show that TRPM4 plays a significant role in the excitability of mPFC layer 2/3 pyramidal neurons by modulating neuronal excitability in a calcium-dependent manner.
Pannexin 1 Transgenic Mice: Human Diseases and Sleep-Wake Function Revision
Authors: Battulin Nariman , Kovalzon Vladimir M. , Korablev Alexey , Serova Irina , Kiryukhina Oxana O. , Pechkova Marta G. , Bogotskoy Kirill A. , Tarasova Olga S. , Panchin Yuri
Published: May 2021

In humans and other vertebrates pannexin protein family was discovered by homology to invertebrate gap junction proteins. Several biological functions were attributed to three vertebrate pannexins members. Six clinically significant independent variants of the PANX1 gene lead to human infertility and oocyte development defects, and the Arg217His variant was associated with pronounced symptoms of primary ovarian failure, severe intellectual disability, sensorineural hearing loss, and kyphosis. At the same time, only mild phenotypes were observed in Panx1 knockout mice. In addition, a passenger mutation was identified in a popular line of Panx1 knockout mice, questioning even those effects. Using CRISPR/Cas9, we created a new line of Panx1 knockout mice and a new line of mice with the clinically significant Panx1 substitution (Arg217His). In both cases, we observed no significant changes in mouse size, weight, or fertility. In addition, we attempted to reproduce a previous study on sleep/wake and locomotor activity functions in Panx1 knockout mice and found that previously reported effects were probably not caused by the Panx1 knockout itself. We consider that the pathological role of Arg217His substitution in Panx1, and some Panx1 functions in general calls for a re-evaluation.
Intermittent Hypobaric Hypoxic Preconditioning Provides Neuroprotection by Increasing Antioxidant Activity, Erythropoietin Expression and Preventing Apoptosis and Astrogliosis in the Brain of Adult Rats Exposed to Acute Severe Hypoxia
Authors: Coimbra-Costa Débora , Garzón Fernando , Alva Norma , Pinto Tiago C. C. , Aguado Fernando , Torrella Joan Ramon , Carbonell Teresa , Rama Ramón
Published: May 2021

Background: Exposure to intermittent hypoxia has been demonstrated to be an efficient tool for hypoxic preconditioning, preventing damage to cells and demonstrating therapeutic benefits. We aimed to evaluate the effects of respiratory intermittent hypobaric hypoxia (IHH) to avoid brain injury caused by exposure to acute severe hypoxia (ASH). Methods: biomarkers of oxidative damage, mitochondrial apoptosis, and transcriptional factors in response to hypoxia were assessed by Western blot and immunohistochemistry in brain tissue. Four groups of rats were used: (1) normoxic (NOR), (2) exposed to ASH (FiO2 7% for 6 h), (3) exposed to IHH for 3 h per day over 8 days at 460 mmHg, and (4) ASH preconditioned after IHH. Results: ASH animals underwent increased oxidative-stress-related parameters, an upregulation in apoptotic proteins and had astrocytes with phenotype forms compatible with severe diffuse reactive astrogliosis. These effects were attenuated and even prevented when the animals were preconditioned with IHH. These changes paralleled the inhibition of NF-κB expression and the increase of erythropoietin (EPO) levels in the brain. Conclusions: IHH exerted neuroprotection against ASH-induced oxidative injury by preventing oxidative stress and inhibiting the apoptotic cascade, which was associated with NF-κB downregulation and EPO upregulation.
Viral Interactions with Adaptor-Protein Complexes: A Ubiquitous Trait among Viral Species
Authors: Strazic Geljic Ivana , Kucan Brlic Paola , Musak Lucija , Karner Dubravka , Ambriović-Ristov Andreja , Jonjic Stipan , Schu Peter , Rovis Tihana Lenac
Published: May 2021

Numerous viruses hijack cellular protein trafficking pathways to mediate cell entry or to rearrange membrane structures thereby promoting viral replication and antagonizing the immune response. Adaptor protein complexes (AP), which mediate protein sorting in endocytic and secretory transport pathways, are one of the conserved viral targets with many viruses possessing AP-interacting motifs. We present here different mechanisms of viral interference with AP complexes and the functional consequences that allow for efficient viral propagation and evasion of host immune defense. The ubiquity of this phenomenon is evidenced by the fact that there are representatives for AP interference in all major viral families, covered in this review. The best described examples are interactions of human immunodeficiency virus and human herpesviruses with AP complexes. Several other viruses, like Ebola, Nipah, and SARS-CoV-2, are pointed out as high priority disease-causative agents supporting the need for deeper understanding of virus-AP interplay which can be exploited in the design of novel antiviral therapies.
Annexins and Membrane Repair Dysfunctions in Muscular Dystrophies
Authors: Croissant Coralie , Carmeille Romain , Brévart Charlotte , Bouter Anthony
Published: May 2021

Muscular dystrophies constitute a group of genetic disorders that cause weakness and progressive loss of skeletal muscle mass. Among them, Miyoshi muscular dystrophy 1 (MMD1), limb girdle muscular dystrophy type R2 (LGMDR2/2B), and LGMDR12 (2L) are characterized by mutation in gene encoding key membrane-repair protein, which leads to severe dysfunctions in sarcolemma repair. Cell membrane disruption is a physiological event induced by mechanical stress, such as muscle contraction and stretching. Like many eukaryotic cells, muscle fibers possess a protein machinery ensuring fast resealing of damaged plasma membrane. Members of the annexins A (ANXA) family belong to this protein machinery. ANXA are small soluble proteins, twelve in number in humans, which share the property of binding to membranes exposing negatively-charged phospholipids in the presence of calcium (Ca2+). Many ANXA have been reported to participate in membrane repair of varied cell types and species, including human skeletal muscle cells in which they may play a collective role in protection and repair of the sarcolemma. Here, we discuss the participation of ANXA in membrane repair of healthy skeletal muscle cells and how dysregulation of ANXA expression may impact the clinical severity of muscular dystrophies.
Regulation of Inflammatory Reaction in Health and Disease
Authors: Fioranelli Massimo , Roccia Maria Grazia , Flavin Dana , Cota Linda
Published: May 2021

Inflammation is a key mechanism for the clearance of infective agents and other inflammatory triggers and is pivotal for the repairing processes of the affected tissues. Inflammation is a multistep process driven by a great number of mediators which regulate specific aspects of the inflammatory response, in agreement with a well-defined chronobiological program. A great number of inflammation-related diseases show a deeply altered immune chronobiology (e.g., COVID-19-related cytokines storm). This aspect highlights the need for a deeper understanding of the inflammatory phenomenon. It is fundamental to study inflammation as a multilevel phenomenon. Of particular interest is the low-grade chronic inflammation, which is an etiological factor of many chronic diseases. Nowadays, the therapeutic approach to low grade chronic inflammation is one of the great challenges of traditional pharmacology. Currently, no drugs specifically designed for the treatment of chronic inflammatory forms are available. Today, bioregulatory systems medicine (BrSM) and low dose medicine (LDM), two pharmacological paradigms grounded in systems medicine, potentially represent new tools for the treatment of inflammation-related diseases. Scientific research has assessed the effectiveness and safety of both these therapeutic approaches, in particular for the management of chronic inflammatory conditions and chronic immunological dysregulations.
Clinical Implications of Exosomes: Targeted Drug Delivery for Cancer Treatment
Authors: Massey Andrew E. , Malik Shabnam , Sikander Mohammad , Doxtater Kyle A. , Tripathi Manish K. , Khan Sheema , Yallapu Murali M. , Jaggi Meena , Chauhan Subhash C. , Hafeez Bilal B.
Published: May 2021

Exosomes are nanoscale vesicles generated by cells for intercellular communication. Due to their composition, significant research has been conducted to transform these particles into specific delivery systems for various disease states. In this review, we discuss the common isolation and loading methods of exosomes, some of the major roles of exosomes in the tumor microenvironment, as well as discuss recent applications of exosomes as drug delivery vessels and the resulting clinical implications.
Cold Atmospheric Plasma Promotes Regeneration-Associated Cell Functions of Murine Cementoblasts In Vitro
Authors: Eggers Benedikt , Marciniak Jana , Deschner James , Stope Matthias Bernhard , Mustea Alexander , Kramer Franz-Josef , Nokhbehsaim Marjan
Published: May 2021

The aim of the study was to examine the efficacy of cold atmospheric plasma (CAP) on the mineralization and cell proliferation of murine dental cementoblasts. Cells were treated with CAP and enamel matrix derivates (EMD). Gene expression of alkaline phosphatase (ALP), bone gamma-carboxyglutamate protein (BGLAP), periostin (POSTN), osteopontin (OPN), osterix (OSX), collagen type I alpha 1 chain (COL1A1), dentin matrix acidic phosphoprotein (DMP)1, RUNX family transcription factor (RUNX)2, and marker of proliferation Ki-67 (KI67) was quantified by real-time PCR. Protein expression was analyzed by immunocytochemistry and ELISA. ALP activity was determined by ALP assay. Von Kossa and alizarin red staining were used to display mineralization. Cell viability was analyzed by XTT assay, and morphological characterization was performed by DAPI/phalloidin staining. Cell migration was quantified with an established scratch assay. CAP and EMD upregulated both mRNA and protein synthesis of ALP, POSTN, and OPN. Additionally, DMP1 and COL1A1 were upregulated at both gene and protein levels. In addition to upregulated RUNX2 mRNA levels, treated cells mineralized more intensively. Moreover, CAP treatment resulted in an upregulation of KI67, higher cell viability, and improved cell migration. Our study shows that CAP appears to have stimulatory effects on regeneration-associated cell functions in cementoblasts.
Regeneration in Spinal Disease: Therapeutic Role of Hypoxia-Inducible Factor-1 Alpha in Regeneration of Degenerative Intervertebral Disc
Authors: Kim Jin-Woo , Jeon Neunghan , Shin Dong-Eun , Lee So-Young , Kim Myongwhan , Han Dong Hun , Shin Jae Yeon , Lee Soonchul
Published: May 2021

The intervertebral disc (IVD) is a complex joint structure comprising three primary components—namely, nucleus pulposus (NP), annulus fibrosus (AF), and cartilaginous endplate (CEP). The IVD retrieves oxygen from the surrounding vertebral body through CEP by diffusion and likely generates ATP via anaerobic glycolysis. IVD degeneration is characterized by a cascade of cellular, compositional, structural changes. With advanced age, pronounced changes occur in the composition of the disc extracellular matrix (ECM). NP and AF cells in the IVD possess poor regenerative capacity compared with that of other tissues. Hypoxia-inducible factor (HIF) is a master transcription factor that initiates a coordinated cellular cascade in response to a low oxygen tension environment, including the regulation of numerous enzymes in response to hypoxia. HIF-1α is essential for NP development and homeostasis and is involved in various processes of IVD degeneration process, promotes ECM in NP, maintains the metabolic activities of NP, and regulates dystrophic mineralization of NP, as well as angiogenesis, autophagy, and apoptosis during IVD degeneration. HIF-1α may, therefore, represent a diagnostic tool for early IVD degeneration and a therapeutic target for inhibiting IVD degeneration
The Taxonomic History of Ochlerotatus Lynch Arribálzaga, 1891 (Diptera: Culicidae)
Authors: Ferreira de Freitas Lílian , Bartholomay Lyric C.
Published: May 2021

A review of all taxonomic actions within the subgenus Ochlerotatus Lynch Arribálzaga, 1891 (Diptera: Culicidae) sensu Reinert et al. (2008) is provided. In particular, the complex historical taxonomic treatment of the type species of this group is dissected and explained in detail. Additionally, current challenges with the definition of the subgenus and its constituents are discussed, as are the requisite steps for a successful revision of the taxon. Going forward, we conclude that a taxonomic revision of the species should include a neotype designation for Ochlerotatus scapularis (Rondani, 1848) from topotypical material. Additionally, we provide a review of the characters and taxa that need to be re-evaluated and well-described in order to stabilize the taxonomy of the subgenus. This effort represents a key step towards a stable nomenclature of the Tribe Aedini.
Morphological, Ecological, and Molecular Divergence of Conogethes pinicolalis from C. punctiferalis (Lepidoptera: Crambidae)
Authors: Jeong Na-Ra , Kim Min-Jee , Kim Sung-Soo , Choi Sei-Woong , Kim Ik-Soo
Published: May 2021

Conogethes pinicolalis has long been considered as a Pinaceae-feeding type of the yellow peach moth, C. punctiferalis, in Korea. In this study, the divergence of C. pinicolalis from the fruit-feeding moth C. punctiferalis was analyzed in terms of morphology, ecology, and genetics. C. pinicolalis differs from C. punctiferalis in several morphological features. Through field observation, we confirmed that pine trees are the host plants for the first generation of C. pinicolalis larvae, in contrast to fruit-feeding C. punctiferalis larvae. We successfully reared C. pinicolalis larvae to adults by providing them pine needles as a diet. From a genetic perspective, the sequences of mitochondrial COI of these two species substantially diverged by an average of 5.46%; moreover, phylogenetic analysis clearly assigned each species to an independent clade. On the other hand, nuclear EF1α showed a lower sequence divergence (2.10%) than COI. Overall, EF1α-based phylogenetic analysis confirmed each species as an independent clade, but a few haplotypes of EF1α indicated incomplete lineage sorting between these two species. In conclusion, our results demonstrate that C. pinicolalis is an independent species according to general taxonomic criteria; however, analysis of the EF1α sequence revealed a short divergence time.
Costs and Benefits of Wax Production in the Larvae of the Ladybeetle Scymnus nubilus
Authors: Pacheco Paulo , Borges Isabel , Branco Beatriz , Lucas Eric , Soares António Onofre
Published: May 2021

BACKGROUND: Larvae of the minute aphidophagous Scymnus nubilus Mulsant (Coleoptera: Coccinellidae) are common predators in apple orchards, covered by a wax layer that might act as a defense mechanism against natural enemies. However, the costs and benefits of protection conferred by wax remain to be assessed. We tested the following hypothesis: there is a trade-off in wax producing ladybeetles between the protection conferred by wax cover and the physiological or behavioral costs associated with its production. We predict that: (1) wax production is an efficient defensive mechanism (against intraguild predation), (2) wax production is associated with detrimental physiological (growth, reproduction) or behavioral effects (behavioral compensation: increased biomass consumption). RESULTS: Tests were carried out in the laboratory with wax and waxless larvae of S. nubilus, with and without lacewing larvae of Chrysoperla agilis (Neuroptera: Chrysopidae) being used as a potential intraguild predator of the coccinellid. Waxless individuals were more susceptible to intraguild predation by lacewing larvae. Adults originating from waxless larvae were lighter than the ones originating from wax larvae, suggesting a metabolic cost resulting from a constant need of wax production. Body-weight gain and conversion efficiency were lower in waxless larvae. Biomass consumption was similar, showing that waxless larvae did not compensate for the physiological cost by eating more aphid biomass. CONCLUSION: The results indicate the potential existence of a trade-off between growth and protection associated with wax production.