bluuebunny/crossref_metadata_2025 huggingface dataset:
The Role of Snack Choices, Body Weight Stereotypes and Smoking Behavior in Assessing Risk Factors for Adolescent Overweight and Obesity
Authors: Raptou Elena
Published: March 2021

This study investigated the relationship of behavioral factors, such as snack choices, obesity stereotypes and smoking with adolescents’ body weight. Individual-level data for 1254 Greek youths were selected via a formal questionnaire. Snack choices seem to be gender specific with girls showing a stronger preference for healthier snacks. Frequent consumption of high-calorie and more filling snacks was found to increase Body Mass Index (BMI) in both genders. Fruit/vegetable snacks were associated with lower body weight in females, whereas cereal/nut snacks had a negative influence in males’ BMI. The majority of participants expressed anti-fat attitudes and more boys than girls assigned positive attributes to lean peers. The endorsement of the thin-ideal was positively associated with the BMI of both adolescent boys and girls. This study also revealed that neglecting potential endogeneity issues can lead to biased estimates of smoking. Gender may be a crucial moderator of smoking–BMI relationships. Male smokers presented a higher obesity risk, whereas female smokers were more likely to be underweight. Nutrition professionals should pay attention to increase the acceptance of healthy snack options. Gender differences in the influence of weight stereotypes and smoking on BMI should be considered in order to enhance the efficacy of obesity prevention interventions.
Fuel Cell Fractional-Order Model via Electrochemical Impedance Spectroscopy
Authors: Caponetto Riccardo , Matera Fabio , Murgano Emanuele , Privitera Emanuela , Xibilia Maria Gabriella
Published: March 2021

The knowledge of the electrochemical processes inside a Fuel Cell (FC) is useful for improving FC diagnostics, and Electrochemical Impedance Spectroscopy (EIS) is one of the most used techniques for electrochemical characterization. This paper aims to propose the identification of a Fractional-Order Transfer Function (FOTF) able to represent the FC behavior in a set of working points. The model was identified by using a data-driven approach. Experimental data were obtained testing a Proton Exchange Membrane Fuel Cell (PEMFC) to measure the cell impedance. A genetic algorithm was firstly used to determine the sets of fractional-order impedance model parameters that best fit the input data in each analyzed working point. Then, a method was proposed to select a single set of parameters, which can represent the system behavior in all the considered working conditions. The comparison with an equivalent circuit model taken from the literature is reported, showing the advantages of the proposed approach.
The Gavorrano Monzogranite (Northern Apennines): An Updated Review of Host Rock Protoliths, Thermal Metamorphism and Tectonic Setting
Authors: Brogi Andrea , Caggianelli Alfredo , Liotta Domenico , Zucchi Martina , Spina Amalia , Capezzuoli Enrico , Casini Alessandra , Buracchi Elena
Published: March 2021

We review and refine the geological setting of an area located nearby the Tyrrhenian seacoast, in the inner zone of the Northern Apennines (southern Tuscany), where a Neogene monzogranite body (estimated in about 3 km long, 1.5 km wide, and 0.7 km thick) emplaced during early Pliocene. This magmatic intrusion, known as the Gavorrano pluton, is partially exposed in a ridge bounded by regional faults delimiting broad structural depressions. A widespread circulation of geothermal fluids accompanied the cooling of the magmatic body and gave rise to an extensive Fe-ore deposit (mainly pyrite) exploited during the past century. The tectonic setting which favoured the emplacement and exhumation of the Gavorrano pluton is strongly debated with fallouts on the comprehension of the Neogene evolution of this sector of the inner Northern Apennines. Data from a new fieldwork dataset, integrated with information from the mining activity, have been integrated to refine the geological setting of the whole crustal sector where the Gavorrano monzogranite was emplaced and exhumed. Our review, implemented by new palynological, petrological and structural data pointed out that: (i) the age of the Palaeozoic phyllite (hosting rocks) is middle-late Permian, thus resulting younger than previously described (i.e., pre-Carboniferous); (ii) the conditions at which the metamorphic aureole developed are estimated at a temperature of c. 660 °C and at a depth lower than c. 6 km; (iii) the tectonic evolution which determined the emplacement and exhumation of the monzogranite is constrained in a transfer zone, in the frame of the extensional tectonics affecting the area continuously since Miocene.
The Impact of COVID-19 on Essential Medicines and Personal Protective Equipment Availability and Prices in Saudi Arabia
Authors: Aljadeed Rana , AlRuthia Yazed , Balkhi Bander , Sales Ibrahim , Alwhaibi Monira , Almohammed Omar , Alotaibi Abdulaziz J. , Alrumaih Ali M. , Asiri Yousif
Published: March 2021

This was a questionnaire-based cross-sectional study that explored the impact of the COVID-19 pandemic on the availability of essential medicine and personal protective equipment (PPE) in Saudi Arabia. Purposive sampling technique was used to recruit individuals working in the supply chain departments in different healthcare sectors in Saudi Arabia. One hundred and three pharmaceutical and medical supply chain employees participated in the study. Most of the participants (58.3%) were aged ≥35 years, male (65%), and pharmacists (92.2%). The majority of participants had at least two years of experience in supply chain (77.6%), worked in public hospitals (95.15%), and were mostly working at healthcare institutions located in Riyadh province (59.2%). Approximately 51% of the participants reported shortages of 10 or more essential drugs. Tocilizumab, hydroxychloroquine, lopinavir/ritonavir, ribavirin, dexamethasone, enoxaparin, interferon beta-1b, cisatracurium besylate, prednisolone, hydrocortisone, methimazole, and methylprednisolone were reported to be in shortage by at least 8% of the participants. Almost 70% of the participants reported that the pandemic did not significantly impact the prices of prescription drugs in shortage (e.g., ≥25%). Moreover, about 70% of the participants reported direct purchasing or procurement of drugs in shortage. Surgical masks, face shields, medical gowns, and N95 respirators were reported to be in short supply by 33% or more of the participants. Approximately 53% of the participants reported the prices of PPE in shortage had seen an increase by at least 25% during the pandemic. Although the COVID-19 pandemic has caused a significant disruption in the global pharmaceutical supply chain, its impact was largely manageable in Saudi healthcare institutions. This can be attributable to multiple reasons such as the effective exchange programs between hospitals and the drastic increase in public healthcare spending to ameliorate the negative impact of the pandemic on the healthcare sector.
Implementation of Cancer Plans in the United States: A Review
Authors: Bacchus Michael W. , McKee Bobbie , Gwede Clement K. , Cogle Christopher R.
Published: March 2021

State cancer plans facilitate prioritization and stakeholder engagement in preventing and controlling cancer. Implementation plans further help stakeholders prioritize efforts, reduce redundancy, and find opportunities for work synergies. A review of cancer plan implementations plans was performed in the development of an implementation plan for the Florida Cancer Plan. This review sought to identify, characterize, and summarize the use of implementation plans that support comprehensive cancer control activities. Although 100% of states and territories published a cancer plan and 78% of states provided funding for implementing their state cancer plans, only 32% published an implementation plan. Commonalities and unique features of state cancer plan implementations are presented and discussed. An example implementation plan is provided for states without a plan to model.
Pricing and Reimbursement Pathways of New Orphan Drugs in South Korea: A Longitudinal Comparison
Authors: Lee Jong Hyuk
Published: March 2021

This study aimed to analyze four current pathways affecting the listing and post-listing prices of new orphan drugs (ODs) in South Korea. These mechanisms were: (1) essential OD, (2) pharmacoeconomic evaluation (PE) waiver OD, (3) weighted average price OD, and (4) PE OD. We analyzed the ratio of the listing price of 48 new ODs to the average adjusted price (AAP) of seven advanced countries and examined the change in the post-listing price. Descriptive statistics were used to analyze the listing and post-listing price changes. The mean and median ratios of the listing price of total new OD to AAP were calculated to be 69.4% and 65.4%, respectively. Essential OD showed the highest mean (93.8%) and median (80.8%) ratios. The mean cumulative price discount rate of the new OD was 7.2% in the third year and 5.7% in the fifth year. The rarity of diseases impacts the listing price of OD, but the political effects of the benefits of OD on the post-listing price of these drugs could not be verified. Further research should be conducted to develop measures that facilitate the practical sharing of budget risks and increase patient access to new ODs.
Lessons from a Surgical Center Satellite Warehouse in a Large Brazilian Public Hospital
Authors: Reis Augusto da Cunha , Oliveira Renata Pereira , Ferreira Letícia Ali Figueiredo , Gomes de Souza Cristina
Published: March 2021

Brazilian public hospitals face several operational problems not only related to poor public management practices and their complex nature, but also the economic, and social contexts. Considering this scenario and the fact that efforts in supply management might affect a hospital organization’s excellence, this research aims to identify improvements in the logistic operations at the surgical center satellite warehouse of a Public Hospital located in Brazil. A case study based on an exploratory and qualitative approach was conducted by employing document analysis, semi-structured interviews, and on-site observations. Seven major problems concerning lack of surgical material, the non-definition of crucial logistic parameters, low information flow, surgical supply control, and management problems were pointed and addressed by seven independent but complementary actions that considered the Brazilian healthcare system’s particularities. Given the nature of exploratory research, the results are not exhaustive and cannot be generalized to different contexts. However, they help understand that reducing the waste of the logistics processes makes it possible to improve the attention to the local population that uses public health services.
Taking the Long View for Oceans and Human Health Connection through Community Driven Science
Authors: Varanasi Usha , Trainer Vera L. , Schumacker Ervin Joe
Published: March 2021

The most proactive approach to resolving current health and climate crises will require a long view, focused on establishing and fostering partnerships to identify and eliminate root causes of the disconnect between humans and nature. We describe the lessons learned through a unique scientific partnership that addresses a specific crisis, harmful algal blooms (HABs), along the northeast Pacific Ocean coast, that blends current-day technology with observational knowledge of Indigenous communities. This integrative scientific strategy resulted in creative solutions for forecasting and managing HAB risk in the Pacific Northwest as a part of the US Ocean and Human Health (OHH) program. Specific OHH projects focused on: (1) understanding genetic responses of tribal members to toxins in the marine environment, (2) knowledge sharing by elders during youth camps; (3) establishing an early warning program to alert resource managers of HABs are explicit examples of proactive strategies used to address environmental problems. The research and monitoring projects with tribal communities taught the collaborating non-Indigenous scientists the value of reciprocity, highlighting both the benefits from and protection of oceans that promote our well-being. Effective global oceans and human health initiatives require a collective action that gives equal respect to all voices to promote forward thinking solutions for ocean health.
Bridging or Bonding: An Organizational Framework for Studying Social Capital in Kindergartens
Authors: Vannebo Berit Irene , Ljunggren Elin Birgitte
Published: March 2021

The article develops our understanding of social capital by analyzing social capital as an organizational phenomenon. The analysis is based on qualitative data consisting of interviews and documents obtained from six different kindergartens in Norway. Kindergartens are used as a “prism” through which we can understand how social capital is formed—and the mechanisms that shape the development of various forms of networks within welfare organizations. More specifically we look at drop-in kindergartens. The specific purpose of these kindergartens is to provide open and inclusive arenas that promote integration and community. We find that the kindergartens vary in the degree to which they succeed in building bridging forms of networks and communities. Using concepts from organizational theory and Wenger’s (1998) theory of communities of practice, we find that formal organizational factors such as ownership, organizational goals, profiling, location, and educational content impact the formation of bridging forms of social capital. The composition of the user groups and the user groups’ motivation for participating most clearly affect the conditions for community formation. The composition of the user groups is the result of a number of organizational factors and organizational mechanisms. Kindergartens that have a heterogeneous user group, and a user group with a community orientation (Morse 2006), are more successful at creating bridging types of social networks.
Three-Dimensional Inundation Mapping Using UAV Image Segmentation and Digital Surface Model
Authors: Gebrehiwot Asmamaw A , Hashemi-Beni Leila
Published: March 2021

Flood occurrence is increasing due to the expansion of urbanization and extreme weather like hurricanes; hence, research on methods of inundation monitoring and mapping has increased to reduce the severe impacts of flood disasters. This research studies and compares two methods for inundation depth estimation using UAV images and topographic data. The methods consist of three main stages: (1) extracting flooded areas and create 2D inundation polygons using deep learning; (2) reconstructing 3D water surface using the polygons and topographic data; and (3) deriving a water depth map using the 3D reconstructed water surface and a pre-flood DEM. The two methods are different at reconstructing the 3D water surface (stage 2). The first method uses structure from motion (SfM) for creating a point cloud of the area from overlapping UAV images, and the water polygons resulted from stage 1 is applied for water point cloud classification. While the second method reconstructs the water surface by intersecting the water polygons and a pre-flood DEM created using the pre-flood LiDAR data. We evaluate the proposed methods for inundation depth mapping over the Town of Princeville during a flooding event during Hurricane Matthew. The methods are compared and validated using the USGS gauge water level data acquired during the flood event. The RMSEs for water depth using the SfM method and integrated method based on deep learning and DEM were 0.34m and 0.26m, respectively.
Momordica cochinchinensis Aril Ameliorates Diet-Induced Metabolic Dysfunction and Non-Alcoholic Fatty Liver by Modulating Gut Microbiota
Authors: Huang Hsiu-Chen , Chen Chiung-Ju , Lai Yu-Heng , Lin Yu-Chun , Chiou Wei-Chung , Lu Hsu-Feng , Chen Ying-Fang , Chen Yu-Hsin , Huang Cheng
Published: March 2021

Obesity and its associated conditions, such as type 2 diabetes mellitus (T2DM) and nonalcoholic fatty liver disease (NAFLD), are a particular worldwide health problem at present. Momordica cochinchinensis (MC) is consumed widely in Southeast Asia. However, whether it has functional effects on fat-induced metabolic syndrome remains unclear. This study was conducted to examine the prevention effect of Momordica cochinchinensis aril (MCA) on obesity, non-alcoholic fatty liver and insulin resistance in mice. MCA protected the mice against high-fat diet (HFD)-induced body weight gain, hyperlipidemia and hyperglycemia, compared with mice that were not treated. MCA inhibited the expansion of adipose tissue and adipocyte hypertrophy. In addition, the insulin sensitivity-associated index that evaluates insulin function was also significantly restored. MCA also regulated the secretion of adipokines in HFD-induced obese mice. Moreover, hepatic fat accumulation and liver damage were reduced, which suggested that fatty liver was prevented by MCA. Furthermore, MCA supplementation suppressed hepatic lipid accumulation by activation of the AMP-activated protein kinase (AMPK) and peroxisome proliferator-activated receptor-alpha (PPAR-alpha) signaling pathway in the human fatty liver HuS-E/2 cell model. Our data indicate that MCA altered the microbial contents of the gut and modulated microbial dysbiosis in the host, and consequently is involved in the prevention of HFD-induced adiposity, insulin resistance and non-alcoholic fatty liver disease.
Prediction and Activity of a Cationic α-Helix Antimicrobial Peptide ZM-804 from Maize
Authors: Hassan Mohamed F. , Qutb Abdelrahman M. , Dong Wubei
Published: March 2021

Antimicrobial peptides (AMPs) are small molecules consisting of less than fifty residues of amino acids. Plant AMPs establish the first barrier of defense in the innate immune system in response to invading pathogens. The purpose of this study was to isolate new AMPs from the Zea mays L. inbred line B73 and investigate their antimicrobial activities and mechanisms against certain essential plant pathogenic bacteria. In silico, the Collection of Anti-Microbial Peptides (CAMPR3), a computational AMP prediction server, was used to screen a cDNA library for AMPs. A ZM-804 peptide, isolated from the Z. mays L. inbred line B73 cDNA library, was predicted as a new cationic AMP with high prediction values. ZM-804 was tested against eleven pathogens of Gram-negative and Gram-positive bacteria and exhibited high antimicrobial activities as determined by the minimal inhibitory concentrations (MICs) and the minimum bactericidal concentrations (MBCs). A confocal laser scanning microscope observation showed that the ZM-804 AMP targets bacterial cell membranes. SEM and TEM images revealed the disruption and damage of the cell membrane morphology of Clavibacter michiganensis subsp. michiganensis and Pseudomonas syringae pv. tomato (Pst) DC3000 caused by ZM-804. In planta, ZM-804 demonstrated antimicrobial activity and prevented the infection of tomato plants by Pst DC3000. Moreover, four virulent phytopathogenic bacteria were prevented from inducing hypersensitive response (HR) in tobacco leaves in response to low ZM-804 concentrations. ZM-804 exhibits low hemolytic activity against mouse red blood cells (RBCs) and is relatively safe for mammalian cells. In conclusion, the ZM-804 peptide has a strong antibacterial activity and provides an alternative tool for plant disease control. Additionally, the ZM-804 peptide is considered a promising candidate for human and animal drug development.
Loss of Nckx3 Exacerbates Experimental DSS-Induced Colitis in Mice through p53/NF-κB Pathway
Authors: Tran Dinh Nam , Go Seon Myeong , Park Seon-Mi , Jung Eui-Man , Jeung Eui-Bae
Published: March 2021

Inflammatory bowel diseases (IBDs) comprises a range of chronic inflammatory conditions of the intestinal tract. The incidence and prevalence of IBDs are increasing worldwide, but the precise etiology of these diseases is not completely understood. Calcium signaling plays a regulatory role in cellular proliferation. Nckx3, a potassium-dependent Na+/Ca2+ exchanger, is not only expressed in the brain but also in the aortic, uterine, and intestinal tissues, which contain abundant smooth muscle cells. This study investigated the role of Nckx3 in intestinal inflammation. Microarray analyses revealed the upregulation of the innate immune response-associated genes in the duodenum of Nckx3 knockout (KO) mice. The Nckx3 KO mice also showed an increase in IBD- and tumorigenesis-related genes. Using dextran sodium sulfate (DSS)-induced experimental colitis mice models, the Nckx3 KO mice showed severe colitis. Furthermore, the pathways involving p53 and NF-κB signaling were significantly upregulated by the absence of Nckx3. Overall, Nckx3 plays a critical role in the innate immune and immune response and may be central to the pathogenesis of IBD.
Transglutaminases Are Active in Perivascular Adipose Tissue
Authors: Orr Alexis N. , Thompson Janice M. , Lyttle Janae M. , Watts Stephanie W.
Published: March 2021

Transglutaminases (TGs) are crosslinking enzymes best known for their vascular remodeling in hypertension. They require calcium to form an isopeptide bond, connecting a glutamine to a protein bound lysine residue or a free amine donor such as norepinephrine (NE) or serotonin (5-HT). We discovered that perivascular adipose tissue (PVAT) contains significant amounts of these amines, making PVAT an ideal model to test interactions of amines and TGs. We hypothesized that transglutaminases are active in PVAT. Real time RT-PCR determined that Sprague Dawley rat aortic, superior mesenteric artery (SMA), and mesenteric resistance vessel (MR) PVATs express TG2 and blood coagulation Factor-XIII (FXIII) mRNA. Consistent with this, immunohistochemical analyses support that these PVATs all express TG2 and FXIII protein. The activity of TG2 and FXIII was investigated in tissue sections using substrate peptides that label active TGs when in a catalyzing calcium solution. Both TG2 and FXIII were active in rat aortic PVAT, SMAPVAT, and MRPVAT. Western blot analysis determined that the known TG inhibitor cystamine reduced incorporation of experimentally added amine donor 5-(biotinamido)pentylamine (BAP) into MRPVAT. Finally, experimentally added NE competitively inhibited incorporation of BAP into MRPVAT adipocytes. Further studies to determine the identity of amidated proteins will give insight into how these enzymes contribute to functions of PVAT and, ultimately, blood pressure.
Antimetastatic and Antitumor Activities of Orally Administered NAX014 Compound in a Murine Model of HER2-Positive Breast Cancer
Authors: Pierpaoli Elisa , Piacenza Francesco , Fiorillo Gaetano , Lombardi Paolo , Orlando Fiorenza , Salvatore Carmela , Geroni Cristina , Provinciali Mauro
Published: March 2021

The natural isoquinoline alkaloid Berberine (BBR) has been shown to possess several therapeutic effects, including anticancer activity. Different BBR derivatives have been designed and synthesized in order to obtain new compounds with enhanced anticancer efficacy. We previously showed that intraperitoneal (IP) administration of the BBR-derived NAX014 compound was able to counteract HER-2 overexpressing mammary tumors onset and progression in transgenic mice. However, the IP administration was found to induce organ toxicity at doses higher than 2.5 mg/Kg. In this study, we evaluated the effect of intragastric (IG) administration of 20 mg/kg of NAX014 on both safety and anticancer efficacy in HER-2/neu transgenic mice. Furthermore, cancer cell dissemination and migration, tumor cell senescence and immunological changes were examined. Our results demonstrated that IG NAX014 administration delayed the onset of mammary tumors with no negative effects on health and survival. NAX014 reduced HER-2 overexpressing BC cells migration in vitro and the frequency of lung metastasis in HER-2/neu transgenic mice. A statistically significant increase of senescence-associated p16 expression was observed in tumors from NAX014-treated mice, and the induction of cell senescence was observed in HER-2 overexpressing BC cells after in vitro treatment with NAX014. Although NAX014 did not modulate the presence of tumor-infiltrating lymphocytes, the level of circulating TNF-α and VEGF was found to be reduced in NAX014-treated mice. The overall results address the NAX014 compound as potential tool for therapeutic strategies against HER-2 overexpressing breast cancer.
Label-Free Multiphoton Microscopy: Much More Than Fancy Images
Authors: Borile Giulia , Sandrin Deborah , Filippi Andrea , Anderson Kurt I. , Romanato Filippo
Published: March 2021

Multiphoton microscopy has recently passed the milestone of its first 30 years of activity in biomedical research. The growing interest around this approach has led to a variety of applications from basic research to clinical practice. Moreover, this technique offers the advantage of label-free multiphoton imaging to analyze samples without staining processes and the need for a dedicated system. Here, we review the state of the art of label-free techniques; then, we focus on two-photon autofluorescence as well as second and third harmonic generation, describing physical and technical characteristics. We summarize some successful applications to a plethora of biomedical research fields and samples, underlying the versatility of this technique. A paragraph is dedicated to an overview of sample preparation, which is a crucial step in every microscopy experiment. Afterwards, we provide a detailed review analysis of the main quantitative methods to extract important information and parameters from acquired images using second harmonic generation. Lastly, we discuss advantages, limitations, and future perspectives in label-free multiphoton microscopy.
Link between Lipid Second Messengers and Osmotic Stress in Plants
Authors: Rodas-Junco Beatriz A. , Racagni-Di-Palma Graciela E. , Canul-Chan Michel , Usorach Javier , Hernández-Sotomayor S. M. Teresa
Published: March 2021

Plants are subject to different types of stress, which consequently affect their growth and development. They have developed mechanisms for recognizing and processing an extracellular signal. Second messengers are transient molecules that modulate the physiological responses in plant cells under stress conditions. In this sense, it has been shown in various plant models that membrane lipids are substrates for the generation of second lipid messengers such as phosphoinositide, phosphatidic acid, sphingolipids, and lysophospholipids. In recent years, research on lipid second messengers has been moving toward using genetic and molecular approaches to reveal the molecular setting in which these molecules act in response to osmotic stress. In this sense, these studies have established that second messengers can transiently recruit target proteins to the membrane and, therefore, affect protein conformation, activity, and gene expression. This review summarizes recent advances in responses related to the link between lipid second messengers and osmotic stress in plant cells.
Spectroscopic Signature of Red Blood Cells in a D-Galactose-Induced Accelerated Aging Model
Authors: Blat Aneta , Stepanenko Tetiana , Bulat Katarzyna , Wajda Aleksandra , Dybas Jakub , Mohaissen Tasnim , Alcicek Fatih Celal , Szczesny-Malysiak Ewa , Malek Kamilla , Fedorowicz Andrzej , Marzec Katarzyna M.
Published: March 2021

This work presents a semi-quantitative spectroscopic approach, including FTIR–ATR and Raman spectroscopies, for the biochemical analysis of red blood cells (RBCs) supported by the biochemical, morphological and rheological reference techniques. This multi-modal approach provided the description of the RBC alterations at the molecular level in a model of accelerated aging induced by administration of D-galactose (D-gal), in comparison to natural aging. Such an approach allowed to conclude that most age-related biochemical RBC membrane changes (a decrease in lipid unsaturation and the level of phospholipids, or an increase in acyl chain shortening) as well as alterations in the morphological parameters and RBC deformability are well reflected in the D-gal model of accelerated aging. Similarly, as in natural aging, a decrease in LDL level in blood plasma and no changes in the fraction of glucose, creatinine, total cholesterol, HDL, iron, or triglycerides were observed during the course of accelerated aging. Contrary to natural aging, the D-gal model led to an increase in cholesterol esters and the fraction of total esterified lipids in RBC membranes, and evoked significant changes in the secondary structure of the membrane proteins. Moreover, a significant decrease in the phosphorous level of blood plasma was specific for the D-gal model. On the other hand, natural aging induced stronger changes in the secondary structures of the proteins of the RBCs’ interior. This work proves that research on the aging mechanism, especially in circulation-related diseases, should employ the D-gal model with caution. Nonetheless, the D-gal model enables to imitate age-related rheological alterations in RBCs, although they are partially derived from different changes observed in the RBC membrane at the molecular level.
Spinal Inhibitory Interneurons: Gatekeepers of Sensorimotor Pathways
Authors: Stachowski Nicholas J. , Dougherty Kimberly J.
Published: March 2021

The ability to sense and move within an environment are complex functions necessary for the survival of nearly all species. The spinal cord is both the initial entry site for peripheral information and the final output site for motor response, placing spinal circuits as paramount in mediating sensory responses and coordinating movement. This is partly accomplished through the activation of complex spinal microcircuits that gate afferent signals to filter extraneous stimuli from various sensory modalities and determine which signals are transmitted to higher order structures in the CNS and to spinal motor pathways. A mechanistic understanding of how inhibitory interneurons are organized and employed within the spinal cord will provide potential access points for therapeutics targeting inhibitory deficits underlying various pathologies including sensory and movement disorders. Recent studies using transgenic manipulations, neurochemical profiling, and single-cell transcriptomics have identified distinct populations of inhibitory interneurons which express an array of genetic and/or neurochemical markers that constitute functional microcircuits. In this review, we provide an overview of identified neural components that make up inhibitory microcircuits within the dorsal and ventral spinal cord and highlight the importance of inhibitory control of sensorimotor pathways at the spinal level.
Additive Role of Immune System Infiltration and Angiogenesis in Uveal Melanoma Progression
Authors: García-Mulero Sandra , Alonso Maria Henar , del Carpio Luis P. , Sanz-Pamplona Rebeca , Piulats Josep M.
Published: March 2021

Uveal melanoma (UM) is a malignant tumor that arises in the melanocytes of the uveal tract. It is the most frequent eye cancer, and despite new therapeutic approaches, prognosis is still poor, with up to 50% of patients developing metastasis with no efficient treatment options available. In contrast to cutaneous melanoma, UM is considered an “immune-cold” tumor due to the low mutational burden and the unique immunosuppressive microenvironment. To gain insight into the role of the UM microenvironment in regard to prognosis and metastatic progression, we have performed a pool analysis characterizing the UM microenvironment by using a bioinformatic approach. A variety of scores based on gene expression measuring stromal infiltration were calculated and used to assess association with prognosis. As a result, the highest immune and stromal scores were associated with poor prognosis. Specifically, stromal cells (fibroblasts and endothelial cells), T cells CD8+, natural killer (NK) cells, and macrophages M1 and M2 infiltration were associated with poor prognosis. Contrary to other tumors, lymphocytic infiltration is related to poor prognosis. Only B cells were associated with more favorable prognosis. UM samples scoring high in both angiogenesis (Angio) and antigen presentation (AP) pathways showed a poor prognosis suggesting an additive role of both functions. Almost all these tumors exhibited a chromosome 3 monosomy. Finally, an enrichment analysis showed that tumors classified as high Angio-high AP also activated metabolic pathways such as glycolysis or PI3K-AKT-MTOR. In summary, our pool analysis identified a cluster of samples with angiogenic and inflammatory phenotypes exhibiting poor prognosis and metabolic activation. Our analysis showed robust results replicated in a pool analysis merging different datasets from different analytic platforms.
Expression Profiles of Alkaloid-Related Genes across the Organs of Narrow-Leafed Lupin (Lupinus angustifolius L.) and in Response to Anthracnose Infection
Authors: Czepiel Katarzyna , Krajewski Paweł , Wilczura Paulina , Bielecka Patrycja , Święcicki Wojciech , Kroc Magdalena
Published: March 2021

The main restraint obstructing the wider adoption of lupins as protein crops is the presence of bitter and toxic quinolizidine alkaloids (QAs), whose contents might increase under exposure to stressful environmental conditions. A poor understanding of how QAs accumulate hinders the breeding of sweet varieties. Here, we characterize the expression profiles of QA-related genes, along with the alkaloid content, in various organs of sweet and bitter narrow-leafed lupin (NLL, Lupinus angustifolius L.). Special attention is paid to the RAP2-7 transcription factor, a candidate regulator of the QA pathway. We demonstrate the upregulation of RAP2-7 and other QA-related genes, across the aerial organs of a bitter cultivar and the significant correlations between their expression levels, thus supporting the role of RAP2-7 as an important regulatory gene in NLL. Moreover, we showed that the initial steps of QA synthesis might occur independently in all aerial plant organs sharing common regulatory mechanisms. Nonetheless, other regulatory steps might be involved in RAP2-7-triggered QA accumulation, given its expression pattern in leaves. Finally, the examination of QA-related gene expression in plants infected with Colletotrichum lupini evidenced no connection between QA synthesis and anthracnose resistance, in contrast to the important role of polyamines during plant–pathogen interactions.
Changes in the Flower and Leaf Proteome of Common Buckwheat (Fagopyrum esculentum Moench) under High Temperature
Authors: Kopeć Przemysław , Hornyák Marta , Pastuszak Jakub , Szczerba Anna , Rapacz Marcin , Waga Jacek , Płażek Agnieszka
Published: March 2021

Common buckwheat (Fagopyrum esculentum Moench), a pseudocereal crop, produces a large number of flowers, but this does not guarantee high seed yields. This species demonstrates strong abortion of flowers and embryos. High temperatures during the generative growth phase result in an increase in the degeneration of embryo sacs. The aim of this study was to investigate proteomic changes in flowers and leaves of two common buckwheat accessions with different degrees of heat tolerance, Panda and PA15. Two-dimensional gel electrophoresis and mass spectrometry techniques were used to analyze the proteome profiles. Analyses were conducted for flower buds, open flowers capable of fertilization, and wilted flowers, as well as donor leaves, i.e., those growing closest to the inflorescences. High temperature up-regulated the expression of 182 proteins. The proteomic response to heat stress differed between the accessions and among their organs. In the Panda accession, we observed a change in abundance of 17, 13, 28, and 11 proteins, in buds, open and wilted flowers, and leaves, respectively. However, in the PA15 accession there were 34, 21, 63, and 21 such proteins, respectively. Fifteen heat-affected proteins were common to both accessions. The indole-3-glycerol phosphate synthase chloroplastic-like isoform X2 accumulated in the open flowers of the heat-sensitive cultivar Panda in response to high temperature, and may be a candidate protein as a marker of heat sensitivity in buckwheat plants.
Generation and Characterization of a Polyclonal Human Reference Antibody to Measure Anti-Drug Antibody Titers in Patients with Fabry Disease
Authors: Lenders Malte , Scharnetzki David , Heidari Ali , Di Iorio Daniele , Wegner Seraphine Valeska , Brand Eva
Published: March 2021

Male patients with Fabry disease (FD) are at high risk for the formation of antibodies to recombinant α-galactosidase A (AGAL), used for enzyme replacement therapy. Due to the rapid disease progression, the identification of patients at risk is highly warranted. However, currently suitable references and standardized protocols for anti-drug antibodies (ADA) determination do not exist. Here we generate a comprehensive patient-derived antibody mixture as a reference, allowing ELISA-based quantification of antibody titers from individual blood samples. Serum samples of 22 male patients with FD and ADAs against AGAL were pooled and purified by immune adsorption. ADA-affinities against agalsidase-α, agalsidase-β and Moss-AGAL were measured by quartz crystal microbalance with dissipation monitoring (QCM-D). AGAL-specific immune adsorption generated a polyclonal ADA mixture showing a concentration-dependent binding and inhibition of AGAL. Titers in raw sera and from purified total IgGs (r2 = 0.9063 and r2 = 0.8952, both p < 0.0001) correlated with the individual inhibitory capacities of ADAs. QCM-D measurements demonstrated comparable affinities of the reference antibody for agalsidase-α, agalsidase-β and Moss-AGAL (KD: 1.94 ± 0.11 µM, 2.46 ± 0.21 µM, and 1.33 ± 0.09 µM, respectively). The reference antibody allows the ELISA-based ADA titer determination and quantification of absolute concentrations. Furthermore, ADAs from patients with FD have comparable affinities to agalsidase-α, agalsidase-β and Moss-AGAL.
Interactions of SARS-CoV-2 with the Blood–Brain Barrier
Authors: Erickson Michelle A. , Rhea Elizabeth M. , Knopp Rachel C. , Banks William A.
Published: March 2021

Emerging data indicate that neurological complications occur as a consequence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. The blood–brain barrier (BBB) is a critical interface that regulates entry of circulating molecules into the CNS, and is regulated by signals that arise from the brain and blood compartments. In this review, we discuss mechanisms by which SARS-CoV-2 interactions with the BBB may contribute to neurological dysfunction associated with coronavirus disease of 2019 (COVID-19), which is caused by SARS-CoV-2. We consider aspects of peripheral disease, such as hypoxia and systemic inflammatory response syndrome/cytokine storm, as well as CNS infection and mechanisms of viral entry into the brain. We also discuss the contribution of risk factors for developing severe COVID-19 to BBB dysfunction that could increase viral entry or otherwise damage the brain.
microRNA-mRNA Profile of Skeletal Muscle Differentiation and Relevance to Congenital Myotonic Dystrophy
Authors: Morton Sarah U. , Sefton Christopher R. , Zhang Huanqing , Dai Manhong , Turner David L. , Uhler Michael D. , Agrawal Pankaj B.
Published: March 2021

microRNAs (miRNAs) regulate messenger RNA (mRNA) abundance and translation during key developmental processes including muscle differentiation. Assessment of miRNA targets can provide insight into muscle biology and gene expression profiles altered by disease. mRNA and miRNA libraries were generated from C2C12 myoblasts during differentiation, and predicted miRNA targets were identified based on presence of miRNA binding sites and reciprocal expression. Seventeen miRNAs were differentially expressed at all time intervals (comparing days 0, 2, and 5) of differentiation. mRNA targets of differentially expressed miRNAs were enriched for functions related to calcium signaling and sarcomere formation. To evaluate this relationship in a disease state, we evaluated the miRNAs differentially expressed in human congenital myotonic dystrophy (CMD) myoblasts and compared with normal control. Seventy-four miRNAs were differentially expressed during healthy human myocyte maturation, of which only 12 were also up- or downregulated in CMD patient cells. The 62 miRNAs that were only differentially expressed in healthy cells were compared with differentiating C2C12 cells. Eighteen of the 62 were conserved in mouse and up- or down-regulated during mouse myoblast differentiation, and their C2C12 targets were enriched for functions related to muscle differentiation and contraction.
From Pathogenesis to Therapy in Knee Osteoarthritis: Bench-to-Bedside
Authors: Rezuş Elena , Burlui Alexandra , Cardoneanu Anca , Macovei Luana Andreea , Tamba Bogdan Ionel , Rezuş Ciprian
Published: March 2021

Osteoarthritis (OA) is currently the most widespread musculoskeletal condition and primarily affects weight-bearing joints such as the knees and hips. Importantly, knee OA remains a multifactorial whole-joint disease, the appearance and progression of which involves the alteration of articular cartilage as well as the synovium, subchondral bone, ligaments, and muscles through intricate pathomechanisms. Whereas it was initially depicted as a predominantly aging-related and mechanically driven condition given its clear association with old age, high body mass index (BMI), and joint malalignment, more recent research identified and described a plethora of further factors contributing to knee OA pathogenesis. However, the pathogenic intricacies between the molecular pathways involved in OA prompted the study of certain drugs for more than one therapeutic target (amelioration of cartilage and bone changes, and synovial inflammation). Most clinical studies regarding knee OA focus mainly on improvement in pain and joint function and thus do not provide sufficient evidence on the possible disease-modifying properties of the tested drugs. Currently, there is an unmet need for further research regarding OA pathogenesis as well as the introduction and exhaustive testing of potential disease-modifying pharmacotherapies in order to structure an effective treatment plan for these patients.
Control of the Autophagy Pathway in Osteoarthritis: Key Regulators, Therapeutic Targets and Therapeutic Strategies
Authors: Valenti Maria Teresa , Dalle Carbonare Luca , Zipeto Donato , Mottes Monica
Published: March 2021

Autophagy is involved in different degenerative diseases and it may control epigenetic modifications, metabolic processes, stem cells differentiation as well as apoptosis. Autophagy plays a key role in maintaining the homeostasis of cartilage, the tissue produced by chondrocytes; its impairment has been associated to cartilage dysfunctions such as osteoarthritis (OA). Due to their location in a reduced oxygen context, both differentiating and mature chondrocytes are at risk of premature apoptosis, which can be prevented by autophagy. AutophagomiRNAs, which regulate the autophagic process, have been found differentially expressed in OA. AutophagomiRNAs, as well as other regulatory molecules, may also be useful as therapeutic targets. In this review, we describe and discuss the role of autophagy in OA, focusing mainly on the control of autophagomiRNAs in OA pathogenesis and their potential therapeutic applications.
PredNTS: Improved and Robust Prediction of Nitrotyrosine Sites by Integrating Multiple Sequence Features
Authors: Nilamyani Andi Nur , Auliah Firda Nurul , Moni Mohammad Ali , Shoombuatong Watshara , Hasan Md Mehedi , Kurata Hiroyuki
Published: March 2021

Nitrotyrosine, which is generated by numerous reactive nitrogen species, is a type of protein post-translational modification. Identification of site-specific nitration modification on tyrosine is a prerequisite to understanding the molecular function of nitrated proteins. Thanks to the progress of machine learning, computational prediction can play a vital role before the biological experimentation. Herein, we developed a computational predictor PredNTS by integrating multiple sequence features including K-mer, composition of k-spaced amino acid pairs (CKSAAP), AAindex, and binary encoding schemes. The important features were selected by the recursive feature elimination approach using a random forest classifier. Finally, we linearly combined the successive random forest (RF) probability scores generated by the different, single encoding-employing RF models. The resultant PredNTS predictor achieved an area under a curve (AUC) of 0.910 using five-fold cross validation. It outperformed the existing predictors on a comprehensive and independent dataset. Furthermore, we investigated several machine learning algorithms to demonstrate the superiority of the employed RF algorithm. The PredNTS is a useful computational resource for the prediction of nitrotyrosine sites. The web-application with the curated datasets of the PredNTS is publicly available.
Roles of Brassinosteroids in Mitigating Heat Stress Damage in Cereal Crops
Authors: Kothari Aishwarya , Lachowiec Jennifer
Published: March 2021

Heat stress causes huge losses in the yield of cereal crops. Temperature influences the rate of plant metabolic and developmental processes that ultimately determine the production of grains, with high temperatures causing a reduction in grain yield and quality. To ensure continued food security, the tolerance of high temperature is rapidly becoming necessary. Brassinosteroids (BR) are a class of plant hormones that impact tolerance to various biotic and abiotic stresses and regulate cereal growth and fertility. Fine-tuning the action of BR has the potential to increase cereals’ tolerance and acclimation to heat stress and maintain yields. Mechanistically, exogenous applications of BR protect yields through amplifying responses to heat stress and rescuing the expression of growth promoters. Varied BR compounds and differential signaling mechanisms across cereals point to a diversity of mechanisms that can be leveraged to mitigate heat stress. Further, hormone transport and BR interaction with other molecules in plants may be critical to utilizing BR as protective agrochemicals against heat stress. Understanding the interplay between heat stress responses, growth processes and hormone signaling may lead us to a comprehensive dogma of how to tune BR application for optimizing cereal growth under challenging environments in the field.