bluuebunny/crossref_metadata_2025 huggingface dataset:
Increasing Seasonal Influenza Vaccination among University Students: A Systematic Review of Programs Using a Social Marketing Perspective
Authors: Lee Daisy , Rundle-Thiele Sharyn , Wut Tai Ming , Li Gabriel
Published: June 2022

The health and economic consequences of seasonal influenza present great costs to communities. Promoting voluntary uptake of the seasonal influenza vaccine among university students, particularly during the COVID-19 pandemic, can deliver protective effects for both individuals and the wider community. Vaccine uptake will be greatest when more of the social marketing benchmarks are applied. This systematic review summarizes evidence from programs aiming to increase seasonal influenza vaccination among university students. Six major electronic databases for health promotion studies (PubMed, EBSCO, ProQuest, Ovid, Web of Science, and ScienceDirect) were searched in November 2021 to capture peer-reviewed studies reporting field trials that have sought to increase seasonal influenza vaccination in university student populations, without any restrictions regarding the publication period. Following PRISMA guidelines, this paper identified 12 peer-reviewed studies that were conducted in the field in the United States, Australia, and Spain. Three studies were targeted at healthcare students and the rest focused on wider university student populations. Studies were narratively summarized, evidence of social marketing principles were identified, and quantitative outcomes were meta-analyzed. The findings indicate that none of the field studies, even a self-classified social marketing study, had adopted all eight of the social marketing benchmarks in program design and implementation. The two studies that only used promotion, but not other marketing-mix and social marketing principles, reported increases in students’ intention to be vaccinated but not actual behavior. Given that change is more likely when more social benchmarks are applied, this paper identifies activities that can be included in flu vaccine programs to improve flu vaccine uptake rates. The analysis highlights a lack of field studies focusing on increasing rates of vaccination behavior as research outcomes in countries beyond the United States.
The GRANDPACT Project: The Development and Evaluation of an Intergenerational Program for Grandchildren and Their Grandparents to Stimulate Physical Activity and Cognitive Function Using Co-Creation
Authors: Iliano Evelien , Beeckman Melanie , Latomme Julie , Cardon Greet
Published: June 2022

In recent years, increased attention has been devoted to intergenerational physical activity (PA) programs because they may have several benefits for both children and older adults (e.g., the reduction of ageism). An intergenerational PA program focusing on grandchildren and grandparents in a ‘standard’ family setting that combines PA and cognitive function is innovative and may hold potential for promoting PA and improving cognitive functioning in both grandchildren and grandparents. The aim of this study is to describe the protocol of the GRANDPACT (GRANDparents and GRANDchildren improve their Physical Activity and Cognitive functions using co-creaTion) Project, focusing on the development of an intergenerational, cognitively enriched, movement program for grandchildren and grandparents using the theoretical framework of the “Behaviour Change Wheel” in combination with a co-creation approach. Two co-creation trajectories will be organized to develop the program, followed by a pilot study to refine the program and an RCT with a pre-test (at baseline), a post-test (after 24 weeks), and a follow-up (after 36 weeks) to measure the outcomes of co-PA, cognitive functions, psychosocial well-being, and the quality of the family relationship ingrandchildren and grandparents. The outcomes will be measured using accelerometry for PA, the Cambridge Neuropsychological Test Automated Battery (CANTAB) for cognitive functions, and questionnaires for the psychological well-being and quality of the family relationship. Co-development with end-users and stakeholders during both co-creation trajectories is expected to result in an effective, attractive, and feasible program. Co-PA is expected to improve PA, cognitive functioning, psychosocial well-being, and the quality of the family relationships between grandchildren and grandparents.
Assessing Fluorosis Incidence in Areas with Low Fluoride Content in the Drinking Water, Fluorotic Enamel Architecture, and Composition Alterations
Authors: Strużycka Izabela , Olszewska Aneta , Bogusławska-Kapała Agnieszka , Hryhorowicz Szymon , Kaczmarek-Ryś Marta , Grabarek Beniamin Oskar , Staszkiewicz Rafał , Kuciel-Polczak Izabela , Czajka-Jakubowska Agata
Published: June 2022

There is currently no consensus among researchers on the optimal level of fluoride for human growth and health. As drinking water is not the sole source of fluoride for humans, and fluoride can be found in many food sources, this work aimed to determine the incidence and severity of dental fluorosis in Poland, in areas where a low fluoride content characterizes the drinking water, and to assess the impact of fluoride on the enamel composition and microstructure. The dental examination involved 696 patients (aged 15–25 years) who had since birth lived in areas where the fluoride concentration in drinking water did not exceed 0.25 mg/L. The severity of the condition was evaluated using the Dean’s Index. Both healthy teeth and teeth with varying degrees of fluorosis underwent laboratory tests designed to assess the total protein and fluoride content of the enamel. Protein amount was assessed spectrophotometrically while the level of fluoride ions was measured by DX-120 ion chromatography. The clinical study revealed 89 cases (12.8%) of dental fluorosis of varying severity. The enamel of teeth with mild and moderate fluorosis contained a significantly higher protein (p-value < 0.001 and 0.002, respectively) and fluoride level (p < 0.001) than those with no clinical signs of fluorosis. SEM images showed irregularities in the structure of the fluorotic enamel. An excessive fluoride level during amelogenesis leads to adverse changes in the chemical composition of tooth enamel and its structure. Moreover, dental fluorosis present in areas where drinking water is low in fluorides indicates a need to monitor the supply of fluoride from other possible sources, regardless of its content in the water.
The Perceived Restorativeness of Outdoor Spatial Characteristics for High School Adolescents: A Case Study from China
Authors: Bai Xiaoxia , Li Xinxin , Yan Ding
Published: June 2022

Heavy schoolwork and overpopulated classrooms have made high schools overstressed environments. Previous investigations have identified a wide body of naturally restorative elements. However, evidence regarding the relationship between spatial typology and its perceived restorativeness (PR) for adolescents is limited. This paper explores the connection between spatial characteristics and PR by linking their restorative quality to how they are actually used. A high school with multiple types of outdoor spaces is used as a case study and typical spatial characteristics (area, distance, and openness) are quantified. A revised perceived restorativeness scale (RPRS) is exploited to assess the restorative quality of different spaces, and a self-reported questionnaire is used to map the actual usage. The obtained results reveal that: (1) the restorativeness of the selected spaces varied considerably, with a natural garden being more restorative than a built environment; (2) the area and openness were positively correlated to the PR, but the distance was negatively correlated; (3) the theoretical dimensions of “getting away” at high school are primarily psychological, not physical; (4) the actual use of outdoor spaces during breaks does not match the students’ favorite places or their PR. These findings expand our understanding of the role of spatial characteristics in PR in high schools and indicate direct links between campus design and restorative quality.
Manganese and Lead Exposure and Early Puberty Onset in Children Living near a Ferromanganese Alloy Plant
Authors: dos Santos Nathália Ribeiro , Rodrigues Juliana Lima Gomes , Bandeira Matheus de Jesus , Anjos Ana Laura dos Santos , Araújo Cecília Freitas da Silva , Adan Luis Fernando Fernandes , Menezes-Filho José Antonio
Published: June 2022

Manganese (Mn) and lead (Pb) have been associated with the deregulation of the neuroendocrine system, which could potentially favor the appearance of precocious puberty (PP) in environmentally exposed children. This study aims to evaluate the exposure to Mn and Pb and their potential effects in anticipating puberty in school-aged children living near a ferromanganese alloy plant in Bahia, Brazil. Toenail, occipital hair and blood samples were collected from 225 school-aged children. Tanner’s scale was used for pubertal staging. Mn in blood (MnB), toenail (MnTn) and hair (MnH) and blood lead (PbB) levels were measured by graphite furnace atomic absorption spectrometry. Puberty-related hormone concentrations were determined by chemiluminescence. The age at which girls’ breasts began to develop was inversely correlated with weight-for-age, height-for-age and BMI-for-age Z-scores (p < 0.05); pubarche also had similar results. Mn biomarker levels did not present differences among pubertal classification nor among children with potential PP or not. Furthermore, Mn exposure was not associated with the age of onset of sexual characteristics for either girls or boys. However, PbB levels were positively correlated with boys’ pubic hair stages (rho = 0.258; p = 0.009) and associated with the age of onset of girls’ pubarche (β = 0.299, 95%CI = 0.055–0.542; p = 0.017). Testosterone and LH concentrations were statistically higher in boys with an increased PbB (p = 0.09 and p = 0.02, respectively). Prospective studies are needed to better assess the association between exposure to Mn and Pb and the early onset of puberty.
The Function of DNA Demethylase Gene ROS1a Null Mutant on Seed Development in Rice (Oryza Sativa) Using the CRISPR/CAS9 System
Authors: Irshad Faiza , Li Chao , Wu Hao-Yu , Yan Yan , Xu Jian-Hong
Published: June 2022

The endosperm is the main nutrient source in cereals for humans, as it is a highly specialized storage organ for starch, lipids, and proteins, and plays an essential role in seed growth and development. Active DNA demethylation regulates plant developmental processes and is ensured by cytosine methylation (5-meC) DNA glycosylase enzymes. To find out the role of OsROS1a in seed development, the null mutant of OsROS1a was generated using the CRISPR/Cas9 system. The null mutant of OsROS1a was stable and heritable, which affects the major agronomic traits, particularly in rice seeds. The null mutant of OsROS1a showed longer and narrower grains, and seeds were deformed containing an underdeveloped and less-starch-producing endosperm with slightly irregularly shaped embryos. In contrast to the transparent grains of the wild type, the grains of the null mutant of OsROS1a were slightly opaque and rounded starch granules, with uneven shapes, sizes, and surfaces. A total of 723 differential expression genes (DEGs) were detected in the null mutant of OsROS1a by RNA-Seq, of which 290 were downregulated and 433 were upregulated. The gene ontology (GO) terms with the top 20 enrichment factors were visualized for cellular components, biological processes, and molecular functions. The key genes that are enriched for these GO terms include starch synthesis genes (OsSSIIa and OsSSIIIa) and cellulose synthesis genes (CESA2, CESA3, CESA6, and CESA8). Genes encoding polysaccharides and glutelin were found to be downregulated in the mutant endosperm. The glutelins were further verified by SDS-PAGE, suggesting that glutelin genes could be involved in the null mutant of OsROS1a seed phenotype and OsROS1a could have the key role in the regulation of glutelins. Furthermore, 378 differentially alternative splicing (AS) genes were identified in the null mutant of OsROS1a, suggesting that the OsROS1a gene has an impact on AS events. Our findings indicated that the function on rice endosperm development in the null mutant of OsROS1a could be influenced through regulating gene expression and AS, which could provide the base to properly understand the molecular mechanism related to the OsROS1a gene in the regulation of rice seed development.
Tissue Engineering and Regenerative Medicine in Pediatric Urology: Urethral and Urinary Bladder Reconstruction
Authors: Casarin Martina , Morlacco Alessandro , Dal Moro Fabrizio
Published: June 2022

In the case of pediatric urology there are several congenital conditions, such as hypospadias and neurogenic bladder, which affect, respectively, the urethra and the urinary bladder. In fact, the gold standard consists of a urethroplasty procedure in the case of urethral malformations and enterocystoplasty in the case of urinary bladder disorders. However, both surgical procedures are associated with severe complications, such as fistulas, urethral strictures, and dehiscence of the repair or recurrence of chordee in the case of urethroplasty, and metabolic disturbances, stone formation, urine leakage, and chronic infections in the case of enterocystoplasty. With the aim of overcoming the issue related to the lack of sufficient and appropriate autologous tissue, increasing attention has been focused on tissue engineering. In this review, both the urethral and the urinary bladder reconstruction strategies were summarized, focusing on pediatric applications and evaluating all the biomaterials tested in both animal models and patients. Particular attention was paid to the capability for tissue regeneration in dependence on the eventual presence of seeded cell and growth factor combinations in several types of scaffolds. Moreover, the main critical features needed for urinary tissue engineering have been highlighted and specifically focused on for pediatric application.
Protease-Activated Receptor 1 in Human Carotid Atheroma Is Significantly Related to Iron Metabolism, Plaque Vulnerability, and the Patient’s Age
Authors: Li Wei , Osman Ehab , Forssell Claes , Yuan Xi-Ming
Published: June 2022

(1) Background: Protease-activated receptor 1 (PAR1) has regulatory functions in inflammation, atherogenesis, and atherothrombosis. Chronic iron administration accelerates arterial thrombosis. Intraplaque hemorrhage and hemoglobin catabolism by macrophages are associated with dysregulated iron metabolism and atherosclerotic lesion instability. However, it remains unknown whether expressions of PAR1 in human atherosclerotic lesions are related to plaque severity, accumulation of macrophages, and iron-related proteins. We investigated the expression of PAR1 and its relation to the expression of ferritin and transferrin receptors in human carotid atherosclerotic plaques and then explored potential connections between their expressions, plaque development, and classical risk factors. (2) Methods: Carotid samples from 39 patients (25 males and 14 females) were immunostained with PAR1, macrophages, ferritin, and transferrin receptor. Double immunocytochemistry of PAR1 and ferritin was performed on THP-1 macrophages exposed to iron. (3) Results: PAR1 expression significantly increases with the patient’s age and the progression of human atherosclerotic plaques. Expressions of PAR1 are significantly correlated with the accumulation of CD68-positive macrophages, ferritin, and transferrin receptor 1 (TfR1), and inversely correlated with levels of high-density lipoprotein. In vitro, PAR1 is significantly increased in macrophages exposed to iron, and the expression of PAR1 is colocalized with ferritin expression. (4) Conclusions: PAR1 is significantly related to the progression of human atherosclerotic lesions and the patient’s age. PAR1 is also associated with macrophage infiltration and accumulation of iron metabolic proteins in human atherosclerotic lesions. Cellular iron-mediated induction of PAR1 and its colocalization with ferritin in macrophages may further indicate an important role of cellular iron in atherothrombosis.
Biomarkers of Dementia with Lewy Bodies: Differential Diagnostic with Alzheimer’s Disease
Authors: Bousiges Olivier , Blanc Frédéric
Published: June 2022

Dementia with Lewy Bodies (DLB) is a common form of cognitive neurodegenerative disease. Only one third of patients are correctly diagnosed due to the clinical similarity mainly with Alzheimer’s disease (AD). In this review, we evaluate the interest of different biomarkers: cerebrospinal fluid (CSF), brain MRI, FP-CIT SPECT, MIBG SPECT, PET by focusing more specifically on differential diagnosis between DLB and AD. FP-CIT SPECT is of high interest to discriminate DLB and AD, but not at the prodromal stage (i.e., MCI). MIBG SPECT with decreased cardiac sympathetic activity, perfusion SPECT with occipital hypoperfusion, FDG PET with occipital hypometabolism and cingulate island signs are of interest at the dementia stage but with a lower validity. Brain MRI has shown differences in group study with lower grey matter concentration of the Insula in prodromal DLB, but its interest in clinical routines is not demonstrated. Concerning CSF biomarkers, many studies have already examined the relevance of AD biomarkers but also alpha-synuclein assays in DLB, so we will focus as comprehensively as possible on other biomarkers (especially those that do not appear to be directly related to synucleinopathy) that may be of interest in the differential diagnosis between AD and DLB. Furthermore, we would like to highlight the growing interest in CSF synuclein RT-QuIC, which seems to be an excellent discrimination tool but its application in clinical routine remains to be demonstrated, given the non-automation of the process.
Towards Anion Recognition and Precipitation with Water-Soluble 1,2,4-Selenodiazolium Salts: Combined Structural and Theoretical Study
Authors: Artemjev Alexey A. , Novikov Anton P. , Burkin Gleb M. , Sapronov Alexander A. , Kubasov Alexey S. , Nenajdenko Valentine G. , Khrustalev Victor N. , Borisov Alexander V. , Kirichuk Anatoly A. , Kritchenkov Andreii S. , Gomila Rosa M. , Frontera Antonio , Tskhovrebov Alexander G.
Published: June 2022

The synthesis and structural characterization of a series of supramolecular complexes of bicyclic cationic pyridine-fused 1,2,4-selenodiazoles with various anions is reported. The binding of trifluoroacetate, tetrachloroaurate, tetraphenylborate, perrhenate, and pertechnetate anions in the solid state is regarded. All the anions interact with selenodiazolium cations exclusively via a pair of “chelating” Se⋯O and H⋯O non-covalent interactions, which make them an attractive, novel, non-classical supramolecular recognition unit or a synthon. Trifluoroacetate salts were conveniently generated via novel oxidation reaction of 2,2′-dipyridyl diselenide with bis(trifluoroacetoxy)iodo)benzene in the presence of corresponding nitriles. Isolation and structural characterization of transient 2-pyridylselenyl trifluoroacetate was achieved. X-ray analysis has demonstrated that the latter forms dimers in the solid state featuring very short and strong Se⋯O and Se⋯N ChB contacts. 1,2,4-Selenodiazolium trifluoroacetates or halides show good solubility in water. In contrast, (AuCl4)−, (ReO4)−, or (TcO4)− derivatives immediately precipitate from aqueous solutions. Structural features of these supramolecular complexes in the solid state are discussed. The nature and energies of the non-covalent interactions in novel assembles were studied by the theoretical methods. To the best of our knowledge, this is the first study that regards perrhenate and pertechnetate as acceptors in ChB interactions. The results presented here will be useful for further developments in anion recognition and precipitation involving cationic 1,2,4-selenodiazoles.
Trastuzumab and Doxorubicin Sequential Administration Increases Oxidative Stress and Phosphorylation of Connexin 43 on Ser368
Authors: Pecoraro Michela , Marzocco Stefania , Franceschelli Silvia , Popolo Ada
Published: June 2022

Human epidermal growth factor receptor-2 (HER2) is overexpressed in up to 30% of breast cancer cases, causing a more aggressive tumour growth and poor prognosis. Trastuzumab, the humanized antibody targeted to HER2, increased the life expectancy of patients, but severe cardiotoxicity emerged as a long-term adverse effect. Clinical evidence highlights that Trastuzumab-induced cardiotoxicity drastically increases in association with Doxorubicin; however, the exact mechanisms involved remain incompletely understood. In order to analyse the molecular mechanisms involved and the possible adaptative responses to Trastuzumab and Doxorubicin treatment, in this study, H9c2 cardiomyoblasts were used. Results showed that Trastuzumab and Doxorubicin sequential administration in cardiomyoblast increased cytosolic and mitochondrial ROS production, intracellular calcium dysregulation, mitochondrial membrane depolarization, and the consequent apoptosis, induced by both Trastuzumab and Doxorubicin alone. Furthermore, in these conditions, we observed increased levels of Connexin43 phosphorylated on Ser368 (pCx43). Since phosphorylation on Ser368 decreases gap junction intracellular communication, thus reducing the spread of death signals to adjacent cells, we hypothesized that the increase in pCx43 could be an adaptative response implemented by cells to defend neighbouring cells by Trastuzumab and Doxorubicin sequential administration. However, the other side of the coin is the resulting conduction abnormalities.
Chronic Hyperkaliemia in Chronic Kidney Disease: An Old Concern with New Answers
Authors: Borrelli Silvio , Matarazzo Ida , Lembo Eugenio , Peccarino Laura , Annoiato Claudia , Scognamiglio Maria Rosaria , Foderini Andrea , Ruotolo Chiara , Franculli Aldo , Capozzi Federica , Yavorskiy Pavlo , Merheb Fatme , Provenzano Michele , La Manna Gaetano , De Nicola Luca , Minutolo Roberto , Garofalo Carlo
Published: June 2022

Increasing potassium intake ameliorates blood pressure (BP) and cardiovascular (CV) prognoses in the general population; therefore the World Health Organization recommends a high-potassium diet (90–120 mEq/day). Hyperkalaemia is a rare condition in healthy individuals due to the ability of the kidneys to effectively excrete dietary potassium load in urine, while an increase in serum K+ is prevalent in patients with chronic kidney disease (CKD). Hyperkalaemia prevalence increases in more advanced CKD stages, and is associated with a poor prognosis. This scenario generates controversy on the correct nutritional approach to hyperkalaemia in CKD patients, considering the unproven link between potassium intake and serum K+ levels. Another concern is that drug-induced hyperkalaemia leads to the down-titration or withdrawal of renin-angiotensin system inhibitors (RASI) and mineralocorticoids receptors antagonists (MRA) in patients with CKD, depriving these patients of central therapeutic interventions aimed at delaying CKD progression and decreasing CV mortality. The new K+-binder drugs (Patiromer and Sodium-Zirconium Cyclosilicate) have proven to be adequate and safe therapeutic options to control serum K+ in CKD patients, enabling RASI and MRA therapy, and possibly, a more liberal intake of fruit and vegetables.
Whole-Genome Identification and Analysis of Multiple Gene Families Reveal Candidate Genes for Theasaponin Biosynthesis in Camellia oleifera
Authors: Yang Liying , Gu Yiyang , Zhou Junqin , Yuan Ping , Jiang Nan , Wu Zelong , Tan Xiaofeng
Published: June 2022

Camellia oleifera is an economically important oilseed tree. Seed meals of C. oleifera have a long history of use as biocontrol agents in shrimp farming and as cleaning agents in peoples’ daily lives due to the presence of theasaponins, the triterpene saponins from the genus Camellia. To characterize the biosynthetic pathway of theasaponins in C. oleifera, members of gene families involved in triterpenoid biosynthetic pathways were identified and subjected to phylogenetic analysis with corresponding members in Arabidopsis thaliana, Camellia sinensis, Actinidia chinensis, Panax ginseng, and Medicago truncatula. In total, 143 triterpenoid backbone biosynthetic genes, 1169 CYP450s, and 1019 UGTs were identified in C. oleifera. The expression profiles of triterpenoid backbone biosynthetic genes were analyzed in different tissue and seed developmental stages of C. oleifera. The results suggested that MVA is the main pathway for triterpenoid backbone biosynthesis. Moreover, the candidate genes for theasaponin biosynthesis were identified by WGCNA and qRT-PCR analysis; these included 11 CYP450s, 14 UGTs, and eight transcription factors. Our results provide valuable information for further research investigating the biosynthetic and regulatory network of theasaponins.
Neuroprotective Effect of Bcl-2 on Lipopolysaccharide-Induced Neuroinflammation in Cortical Neural Stem Cells
Authors: Park Shin-Young , Han Joong-Soo
Published: June 2022

Neuroinflammation is involved in the pathogenesis of neurodegenerative diseases due to increased levels of pro-inflammatory cytokines in the central nervous system (CNS). Chronic neuroinflammation induced by neurotoxic molecules accelerates neuronal damage. B-cell lymphoma 2 (Bcl-2) is generally accepted to be an important anti-apoptotic factor. However, the role of Bcl-2 in neuroprotection against neuroinflammation remains to be determined. The purpose of this study was to investigate the neuroprotective effect of Bcl-2 on lipopolysaccharide (LPS)-induced neuroinflammation in cortical neural stem cells (NSCs). LPS decreased mRNA and protein levels of Tuj-1, a neuron marker, and also suppressed neurite outgrowth, indicating that LPS results in inhibition of neuronal differentiation of NSCs. Furthermore, LPS treatment inhibited Bcl-2 expression during neuronal differentiation; inhibition of neuronal differentiation by LPS was rescued by Bcl-2 overexpression. LPS-induced pro-inflammatory cytokines, including interleukin (IL)-6 and tumor necrosis factor alpha (TNF-α), were decreased by Bcl-2 overexpression. Conversely, Bcl-2 siRNA increased the LPS-induced levels of IL-6 and TNF-α, and decreased neuronal differentiation of NSCs, raising the possibility that Bcl-2 mediates neuronal differentiation by inhibiting the LPS-induced inflammatory response in NSC. These results suggest that Bcl-2 has a neuroprotective effect by inhibiting the LPS-induced inflammatory response in NSCs.
Targeting of Glycosaminoglycans in Genetic and Inflammatory Airway Disease
Authors: Caird Robin , Williamson Michael , Yusuf Azeez , Gogoi Debananda , Casey Michelle , McElvaney Noel G. , Reeves Emer P.
Published: June 2022

In the lung, glycosaminoglycans (GAGs) are dispersed in the extracellular matrix (ECM) occupying the interstitial space between the capillary endothelium and the alveolar epithelium, in the sub-epithelial tissue and in airway secretions. In addition to playing key structural roles, GAGs contribute to a number of physiologic processes ranging from cell differentiation, cell adhesion and wound healing. Cytokine and chemokine–GAG interactions are also involved in presentation of inflammatory molecules to respective receptors leading to immune cell migration and airway infiltration. More recently, pathophysiological roles of GAGs have been described. This review aims to discuss the biological roles and molecular interactions of GAGs, and their impact in the pathology of chronic airway diseases, such as cystic fibrosis and chronic obstructive pulmonary disease. Moreover, the role of GAGs in respiratory disease has been heightened by the current COVID-19 pandemic. This review underlines the essential need for continued research aimed at exploring the contribution of GAGs in the development of inflammation, to provide a better understanding of their biological impact, as well as leads in the development of new therapeutic agents.
Molecular Effects of Mutations in Human Genetic Diseases
Authors: Leonardi Emanuela , Savojardo Castrense , Minervini Giovanni
Published: June 2022

Next-generation sequencing (NGS) has enormously improved the identification of disease-candidate genetic variants [...]
Physiological, Agronomical, and Proteomic Studies Reveal Crucial Players in Rice Nitrogen Use Efficiency under Low Nitrogen Supply
Authors: Tantray Aadil Yousuf , Hazzazi Yehia , Ahmad Altaf
Published: June 2022

Excessive use of nitrogenous fertilizers to enhance rice productivity has become a significant source of nitrogen (N) pollution and reduced sustainable agriculture. However, little information about the physiology of different growth stages, agronomic traits, and associated genetic bases of N use efficiency (NUE) are available at low-N supply. Two rice (Oryza sativa L.) cultivars were grown with optimum N (120 kg ha−1) and low N (60 kg ha−1) supply. Six growth stages were analyzed to measure the growth and physiological traits, as well as the differential proteomic profiles, of the rice cultivars. Cultivar Panvel outclassed Nagina 22 at low-N supply and exhibited improved growth and physiology at most of the growth stages and agronomic efficiency due to higher N uptake and utilization at low-N supply. On average, photosynthetic rate, chlorophyll content, plant biomass, leaf N content, and grain yield were decreased in cultivar Nagina 22 than Panvel was 8%, 11%, 21%, 19%, and 22%, respectively, under low-N supply. Furthermore, proteome analyses revealed that many proteins were upregulated and downregulated at the different growth stages under low-N supply. These proteins are associated with N and carbon metabolism and other physiological processes. This supports the genotypic differences in photosynthesis, N assimilation, energy stabilization, and rice-protein yield. Our study suggests that enhancing NUE at low-N supply demands distinct modifications in N metabolism and physiological assimilation. The NUE may be regulated by key identified differentially expressed proteins. These proteins might be the targets for improving crop NUE at low-N supply.
Comparing α-Quartz-Induced Cytotoxicity and Interleukin-8 Release in Pulmonary Mono- and Co-Cultures Exposed under Submerged and Air-Liquid Interface Conditions
Authors: Friesen Alexandra , Fritsch-Decker Susanne , Hufnagel Matthias , Mülhopt Sonja , Stapf Dieter , Hartwig Andrea , Weiss Carsten
Published: June 2022

The occupational exposure to particles such as crystalline quartz and its impact on the respiratory tract have been studied extensively in recent years. For hazard assessment, the development of physiologically more relevant in-vitro models, i.e., air-liquid interface (ALI) cell cultures, has greatly progressed. Within this study, pulmonary culture models employing A549 and differentiated THP-1 cells as mono-and co-cultures were investigated. The different cultures were exposed to α-quartz particles (Min-U-Sil5) with doses ranging from 15 to 66 µg/cm2 under submerged and ALI conditions and cytotoxicity as well as cytokine release were analyzed. No cytotoxicity was observed after ALI exposure. Contrarily, Min-U-Sil5 was cytotoxic at the highest dose in both submerged mono- and co-cultures. A concentration-dependent release of interleukin-8 was shown for both exposure types, which was overall stronger in co-cultures. Our findings showed considerable differences in the toxicological responses between ALI and submerged exposure and between mono- and co-cultures. A substantial influence of the presence or absence of serum in cell culture media was noted as well. Within this study, the submerged culture was revealed to be more sensitive. This shows the importance of considering different culture and exposure models and highlights the relevance of communication between different cell types for toxicological investigations.
Comparison of Biological Features of Wild European Rabbit Mesenchymal Stem Cells Derived from Different Tissues
Authors: Calle Alexandra , Zamora-Ceballos María , Bárcena Juan , Blanco Esther , Ramírez Miguel Ángel
Published: June 2022

Although the European rabbit is an “endangered” species and a notorious biological model, the analysis and comparative characterization of new tissue sources of rabbit mesenchymal stem cells (rMSCs) have not been well addressed. Here, we report for the first time the isolation and characterization of rMSCs derived from an animal belonging to a natural rabbit population within the native region of the species. New rMSC lines were isolated from different tissues: oral mucosa (rOM-MSC), dermal skin (rDS-MSC), subcutaneous adipose tissue (rSCA-MSC), ovarian adipose tissue (rOA-MSC), oviduct (rO-MSC), and mammary gland (rMG-MSC). The six rMSC lines showed plastic adhesion with fibroblast-like morphology and were all shown to be positive for CD44 and CD29 expression (characteristic markers of MSCs), and negative for CD34 or CD45 expression. In terms of pluripotency features, all rMSC lines expressed NANOG, OCT4, and SOX2. Furthermore, all rMSC lines cultured under osteogenic, chondrogenic, and adipogenic conditions showed differentiation capacity. In conclusion, this study describes the isolation and characterization of new rabbit cell lines from different tissue origins, with a clear mesenchymal pattern. We show that rMSC do not exhibit differences in terms of morphological features, expression of the cell surface, and intracellular markers of pluripotency and in vitro differentiation capacities, attributable to their tissue of origin.
A NAC Transcription Factor from ‘Sea Rice 86′ Enhances Salt Tolerance by Promoting Hydrogen Sulfide Production in Rice Seedlings
Authors: Sun Yan , Song Kaiqiang , Guo Miaomiao , Wu Hao , Ji Xuan , Hou Lixia , Liu Xin , Lu Songchong
Published: June 2022

Soil salinity severely threatens plant growth and crop performance. Hydrogen sulfide (H2S), a plant signal molecule, has been implicated in the regulation of plant responses to salinity stress. However, it is unclear how the transcriptional network regulates H2S biosynthesis during salt stress response. In this study, we identify a rice NAC (NAM, ATAF and CUC) transcription factor, OsNAC35-like (OsNACL35), from a salt-tolerant cultivar ‘Sea Rice 86′ (SR86) and further show that it may have improved salt tolerance via enhanced H2S production. The expression of OsNACL35 was significantly upregulated by high salinity and hydrogen peroxide (H2O2). The OsNACL35 protein was localized predominantly in the nucleus and was found to have transactivation activity in yeast. The overexpression of OsNACL35 (OsNACL35-OE) in japonica cultivar Nipponbare ramatically increased resistance to salinity stress, whereas its dominant-negative constructs (SUPERMAN repression domain, SRDX) conferred hypersensitivity to salt stress in the transgenic lines at the vegetative stage. Moreover, the quantitative real-time PCR analysis showed that many stress-associated genes were differentially expressed in the OsNACL35-OE and OsNACL35-SRDX lines. Interestingly, the ectopic expression of OsNACL35 triggered a sharp increase in H2S content by upregulating the expression of a H2S biosynthetic gene, OsDCD1, upon salinity stress. Furthermore, the dual luciferase and yeast one-hybrid assays indicated that OsNACL35 directly upregulated the expression of OsDCD1 by binding to the promoter sequence of OsDCD1. Taken together, our observations illustrate that OsNACL35 acts as a positive regulator that links H2S production to salt stress tolerance, which may hold promising utility in breeding salt-tolerant rice cultivar.
New Insights into the Determinants of Specificity in Human Type I Arginase: Generation of a Mutant That Is Only Active with Agmatine as Substrate
Authors: Orellana María-Soledad , Jaña Gonzalo A. , Figueroa Maximiliano , Martínez-Oyanedel José , Medina Fabiola E. , Tarifeño-Saldivia Estefanía , Gatica Marcell , García-Robles María Ángeles , Carvajal Nelson , Uribe Elena
Published: June 2022

Arginase catalyzes the hydrolysis of L-arginine into L-ornithine and urea. This enzyme has several analogies with agmatinase, which catalyzes the hydrolysis of agmatine into putrescine and urea. However, this contrasts with the highlighted specificity that each one presents for their respective substrate. A comparison of available crystal structures for arginases reveals an important difference in the extension of two loops located in the entrance of the active site. The first, denominated loop A (I129-L140) contains the residues that interact with the alpha carboxyl group or arginine of arginase, and the loop B (D181-P184) contains the residues that interact with the alpha amino group of arginine. In this work, to determine the importance of these loops in the specificity of arginase, single, double, and triple arginase mutants in these loops were constructed, as well as chimeras between type I human arginase and E. coli agmatinase. In previous studies, the substitution of N130D in arginase (in loop A) generated a species capable of hydrolyzing arginine and agmatine. Now, the specificity of arginase is completely altered, generating a chimeric species that is only active with agmatine as a substrate, by substituting I129T, N130Y, and T131A together with the elimination of residues P132, L133, and T134. In addition, Quantum Mechanic/Molecular Mechanic (QM/MM) calculations were carried out to study the accommodation of the substrates in in the active site of this chimera. With these results it is concluded that this loop is decisive to discriminate the type of substrate susceptible to be hydrolyzed by arginase. Evidence was also obtained to define the loop B as a structural determinant for substrate affinity. Concretely, the double mutation D181T and V182E generate an enzyme with an essentially unaltered kcat value, but with a significantly increased Km value for arginine and a significant decrease in affinity for its product ornithine.
Ischemic Brain Neurodegeneration
Authors: Pluta Ryszard
Published: June 2022

Aging is an inevitable phenomenon experienced by animals and humans, and its intensity varies from person to person [...]
Editorial to the Special Issue: “Dysregulation of Human Molecular and Metabolic Mechanisms Resulting in Oxidative Stress and Damage Generation in the Space Environment”
Authors: Goodwin Thomas J. , Christofidou-Solomidou Melpo
Published: June 2022

Commercial space industries are emergent, bolstered by new exciting rocket systems, orbital and landing vehicles, the creation of multi-country orbital platforms, satellite technology, the renewed promise of low Earth orbit (LEO) business opportunities, as well as promised planetary exploration [...]
Neurosecretory Protein GL Promotes Normotopic Fat Accumulation in Male ICR Mice
Authors: Narimatsu Yuki , Matsuura Daichi , Iwakoshi-Ukena Eiko , Furumitsu Megumi , Ukena Kazuyoshi
Published: June 2022

Neurosecretory protein GL (NPGL) is a small secretory protein identified in the hypothalamus of birds and mammals. We recently reported that NPGL exerts obesogenic effects in obesity-prone C57BL6/J mice. However, whether NPGL elicits adiposity in different mouse strains is poorly understood. In this study, we generated transgenic mice overexpressing Npgl using the ICR strain (Npgl Tg mice) to elucidate the obesogenic effects of NPGL in different strains. Npgl Tg mice showed increased white adipose tissue (WAT) mass. Although the mass of brown adipose tissue (BAT) was slightly altered in Npgl Tg mice, hypertrophy of lipid droplets was also observed in BAT. In contrast, fat accumulation was not induced in the liver, with the upregulation of mRNAs related to hepatic lipolysis. These results support the hypothesis that NPGL causes obesity in several strains and species. This report highlights the pivotal role of NPGL in fat accumulation in adipose tissues and contributes to the elucidation of the biological mechanisms underlying obesity and metabolic diseases in heterogeneous populations.
EGFR-Mutated Non-Small Cell Lung Cancer and Resistance to Immunotherapy: Role of the Tumor Microenvironment
Authors: Madeddu Clelia , Donisi Clelia , Liscia Nicole , Lai Eleonora , Scartozzi Mario , Macciò Antonio
Published: June 2022

Lung cancer is a leading cause of cancer-related deaths worldwide. About 10–30% of patients with non-small cell lung cancer (NSCLC) harbor mutations of the EGFR gene. The Tumor Microenvironment (TME) of patients with NSCLC harboring EGFR mutations displays peculiar characteristics and may modulate the antitumor immune response. EGFR activation increases PD-L1 expression in tumor cells, inducing T cell apoptosis and immune escape. EGFR-Tyrosine Kinase Inhibitors (TKIs) strengthen MHC class I and II antigen presentation in response to IFN-γ, boost CD8+ T-cells levels and DCs, eliminate FOXP3+ Tregs, inhibit macrophage polarization into the M2 phenotype, and decrease PD-L1 expression in cancer cells. Thus, targeted therapy blocks specific signaling pathways, whereas immunotherapy stimulates the immune system to attack tumor cells evading immune surveillance. A combination of TKIs and immunotherapy may have suboptimal synergistic effects. However, data are controversial because activated EGFR signaling allows NSCLC cells to use multiple strategies to create an immunosuppressive TME, including recruitment of Tumor-Associated Macrophages and Tregs and the production of inhibitory cytokines and metabolites. Therefore, these mechanisms should be characterized and targeted by a combined pharmacological approach that also concerns disease stage, cancer-related inflammation with related systemic symptoms, and the general status of the patients to overcome the single-drug resistance development.
Targeted Analysis of HSP70 Isoforms in Human Spermatozoa in the Context of Capacitation and Motility
Authors: Grassi Sarah , Bisconti Marie , Martinet Baptiste , Arcolia Vanessa , Simon Jean-François , Wattiez Ruddy , Leroy Baptiste , Hennebert Elise
Published: June 2022

HSP70s constitute a family of chaperones, some isoforms of which appear to play a role in sperm function. Notably, global proteomic studies analyzing proteins deregulated in asthenozoospermia, a main cause of male infertility characterized by low sperm motility, showed the dysregulation of some HSP70 isoforms. However, to date, no clear trend has been established since the variations in the abundance of HSP70 isoforms differed between studies. The HSPA2 isoform has been reported to play a key role in fertilization, but its dysregulation and possible relocation during capacitation, a maturation process making the spermatozoon capable of fertilizing an oocyte, is debated in the literature. The aim of the present study was to investigate the fate of all sperm HSP70 isoforms during capacitation and in relation to sperm motility. Using Multiple-Reaction Monitoring (MRM) mass spectrometry, we showed that the relative abundance of all detected isoforms was stable between non-capacitated and capacitated spermatozoa. Immunofluorescence using two different antibodies also demonstrated the stability of HSP70 isoform localization during capacitation. We also investigated spermatozoa purified from 20 sperm samples displaying various levels of total and progressive sperm motility. We showed that the abundance of HSP70 isoforms is not correlated to sperm total or progressive motility.
The Role of Heparan Sulfate in CCL26-Induced Eosinophil Chemotaxis
Authors: Pum Alexandra , Ennemoser Maria , Gerlza Tanja , Kungl Andreas J.
Published: June 2022

Proinflammatory chemokine ligand 26 (CCL26, eotaxin-3) mediates transendothelial cell migration of eosinophils by binding and activating the G-protein-coupled (GPC) chemokine receptor 3 on the surface of eosinophilic cells. Here we have investigated the role of glycosaminoglycans (GAGs) as potential co-receptors in the process of CCL26-induced eosinophil chemotaxis. For this purpose, we have first identified the GAG-binding site of CCL26 by a site-directed mutagenesis approach in the form of an alanine screening. A panel of GAG-binding-deficient mutants has been designed, generated, and analyzed with respect to their binding affinities to heparan sulphate (HS) by isothermal fluorescence titration studies. This showed that basic amino acids in the α-helical part of CCL26 are strongly involved in GAG-binding. In chemotaxis experiments, we found that decreased GAG-binding affinity correlated with decreased chemotactic activity, which indicates an involvement of GAGs in eosinophil migration. This was further proven by the negative impact of heparinase III treatment and, independently, by the incubation of eosinophils with an anti heparan sulfate antibody. We finally investigated eosinophils’ proteoglycan (PG) expression patterns by real-time PCR, which revealed the highest expression level for serglycin. Including an anti-serglycin antibody in CCL26-induced eosinophil migration experiments reduced the chemotaxis of these immune cells, thereby proving the dependence of eosinophil mobilization on the proteoglycan serglycin.
Enterococcus and COVID-19: The Emergence of a Perfect Storm?
Authors: Toc Dan Alexandru , Mihaila Razvan Marian , Botan Alexandru , Bobohalma Carina Nicoleta , Risteiu Giulia Andreea , Simut-Cacuci Bogdan Nicolae , Steorobelea Bianca , Troanca Stefan , Junie Lia Monica
Published: June 2022

(1) Background: Based on the uncontrolled use of antibiotics and the lack of worldwide-accepted healthcare policies, the COVID-19 pandemic has provided the best premises for the emergence of life-threatening infections. Based on changes described in the intestinal microbiome, showing an increased number of Enterococcus bacteria and increased intestinal permeability due to viral infection, infections with Enterococcus have taken the spotlight in the healthcare setting; (2) Methods: We conducted a brief review in order to analyze the relationship between the two pathogens: the SARS-CoV-2 virus and the Enterococcus bacterial genus. We searched in PubMed, the Cochrane Library electronic database and MedNar and included twenty-one studies based on relevance; (3) Results: The existing studies show a statistically significant difference in the composition of the intestinal microbiome, favoring Enterococcus genus, when compared to a control group. Changes also seem to persist over a period of time, suggesting possible implications for long COVID. Regarding bloodstream infections, Enterococcus is statistically significantly isolated more often when compared to the pre-COVID-19 era, and to a control group of non-COVID-19 patients. (4) Conclusions: The intimate synergy between COVID-19 and Enterococcus has the potential to pose a real threat to human healthcare, and more extensive research is needed to explore the relationship between these two pathogens.