bluuebunny/crossref_metadata_2025 huggingface dataset:
Long-Term Psychological Effects of COVID-19 Pandemic on Children in Jordan
Authors: Al-Rahamneh Harran , Arafa Lubna , Al Orani Anas , Baqleh Rahaf
Published: July 2021

Millions of children and adolescents have been affected worldwide by quarantine, school closures, and social distancing measures which have been implemented by many countries to control the spread of COVID-19. However, the long-term consequences of such procedures on children’s well-being are not clear. Therefore, this study investigated the psychological impacts of COVID-19 on Jordanian children between the ages of 5–11 years old. A total of 1309 parents with children between the ages of 5 and 11 years old filled in an online survey that included a set of questions to measure their children’s behaviour and emotions before and during the COVID-19 pandemic. Being bored (77.5%), irritable (66%), likely to argue with the rest of the family (60.7%), nervous (54.8%), reluctant (54.2%), and lonely (52.4%) were the most frequently reported symptoms compared to the pre-COVID-19 period. Parents reported that screen use of ≥120 min a day was shown among 48.9% of children and 42% of children did <30 min a day of physical activity. ≤8 h of sleep per night was reported among 42.5% of children compared to pre COVID-19. The results emphasized the importance of developing preventative psychological programs to minimize the impact of the COVID-19 pandemic on children’s psychological well-being.
Effectiveness of Simulation-Based Empathy Enhancement Program for Caregivers (SEE-C) Evaluated by Older Adults Receiving Care
Authors: Lee Kyuwon , Han Areum , Kim Tae Hui
Published: July 2021

The aim of this study was to examine whether a Simulation-based Empathy Enhancement program for Caregivers of the Elderly (SEE-C) was effective in increasing program satisfaction and positive emotional changes of older adults. A total of 100 older adults living alone were randomly assigned to experimental and control groups. The experimental group was interviewed by caregivers who experienced SEE-C while the control group was interviewed by caregivers who did not experience SEE-C. In both elderly groups, post session satisfaction and affective state were assessed using a Session Evaluation Questionnaire (SEQ). Chi-square test and Mann-Whitney U test were conducted. The experimental group (n = 49) reported significantly higher scores than the control group (n = 51) for all three categories of SEQ: session-depth (Mann-Whitney U = 1651.5, p = 0.005), session-smoothness (Mann-Whitney U = 1803.0, p = 0.000), and emotion-positivity (Mann-Whitney U = 1783.0, p = 0.000). However, the experimental group had significantly lower scores for the arousal category of SEQ (Mann-Whitney U = 873.5, p = 0.009). SEE-C could have a positive impact on interviews for elderly care in terms of raising the satisfaction of the interviewee.
Oxidative Stress in Non-Dialysis-Dependent Chronic Kidney Disease Patients
Authors: Tomás-Simó Patricia , D’Marco Luis , Romero-Parra María , Tormos-Muñoz Mari Carmen , Sáez Guillermo , Torregrosa Isidro , Estañ-Capell Nuria , Miguel Alfonso , Gorriz José Luis , Puchades María Jesús
Published: July 2021

Background: Cardiovascular complications are the leading cause of morbidity and mortality at any stage of chronic kidney disease (CKD). Moreover, the high rate of cardiovascular mortality observed in these patients is associated with an accelerated atherosclerosis process that likely starts at the early stages of CKD. Thus, traditional and non-traditional or uremic-related factors represent a link between CKD and cardiovascular risk. Among non-conventional risk factors, particular focus has been placed on anaemia, mineral and bone disorders, inflammation, malnutrition and oxidative stress and, in this regard, connections have been reported between oxidative stress and cardiovascular disease in dialysis patients. Methods: We evaluated the oxidation process in different molecular lines (proteins, lipids and genetic material) in 155 non-dialysis patients at different stages of CKD and 45 healthy controls. To assess oxidative stress status, we analyzed oxidized glutathione (GSSG), reduced glutathione (GSH) and the oxidized/reduced glutathione ratio (GSSG/GSH) and other oxidation indicators, including malondialdehyde (MDA) and 8-oxo-2’-deoxyguanosine (8-oxo-dG). Results: An active grade of oxidative stress was found from the early stages of CKD onwards, which affected all of the molecular lines studied. We observed a heightened oxidative state (indicated by a higher level of oxidized molecules together with decreased levels of antioxidant molecules) as kidney function declined. Furthermore, oxidative stress-related alterations were significantly greater in CKD patients than in the control group. Conclusions: CKD patients exhibit significantly higher oxidative stress than healthy individuals, and these alterations intensify as eGFR declines, showing significant differences between CKD stages. Thus, future research is warranted to provide clearer results in this area.
Effect of Environmental Pollutants PM2.5, CO, NO2, and O3 on the Incidence and Mortality of SARS-CoV-2 Infection in Five Regions of the USA
Authors: Meo Sultan Ayoub , Abukhalaf Abdulelah Adnan , Alessa Omar Mohammed , Alarifi Abdulrahman Saad , Sami Waqas , Klonoff David C.
Published: July 2021

In recent decades, environmental pollution has become a significant international public problem in developing and developed nations. Various regions of the USA are experiencing illnesses related to environmental pollution. This study aims to investigate the association of four environmental pollutants, including particulate matter (PM2.5), carbon monoxide (CO), Nitrogen dioxide (NO2), and Ozone (O3), with daily cases and deaths resulting from SARS-CoV-2 infection in five regions of the USA, Los Angeles, New Mexico, New York, Ohio, and Florida. The daily basis concentrations of PM2.5, CO, NO2, and O3 were documented from two metrological websites. Data were obtained from the date of the appearance of the first case of (SARS-CoV-2) in the five regions of the USA from 13 March to 31 December 2020. Regionally (Los Angeles, New Mexico, New York, Ohio, and Florida), the number of cases and deaths increased significantly along with increasing levels of PM2.5, CO, NO2 and O3 (p < 0.05), respectively. The Poisson regression results further depicted that, for each 1 unit increase in PM2.5, CO, NO2 and O3 levels, the number of SARS-CoV-2 infections significantly increased by 0.1%, 14.8%, 1.1%, and 0.1%, respectively; for each 1 unit increase in CO, NO2, and O3 levels, the number of deaths significantly increased by 4.2%, 3.4%, and 1.5%, respectively. These empirical estimates demonstrate an association between the environmental pollutants PM2.5, CO, NO2, and O3 and SARS-CoV-2 infections, showing that they contribute to the incidence of daily cases and daily deaths in the five different regions of the USA. These findings can inform health policy decisions about combatting the COVID-19 pandemic outbreak in these USA regions and internationally by supporting a reduction in environmental pollution.
Low Horizontal Force Production Capacity during Sprinting as a Potential Risk Factor of Hamstring Injury in Football
Authors: Edouard Pascal , Lahti Johan , Nagahara Ryu , Samozino Pierre , Navarro Laurent , Guex Kenny , Rossi Jérémy , Brughelli Matt , Mendiguchia Jurdan , Morin Jean-Benoît
Published: July 2021

Clear decreases in horizontal force production capacity during sprint acceleration have been reported after hamstring injuries (HI) in football players. We hypothesized that lower FH0 is associated with a higher HI occurrence in football players. We aimed to analyze the association between sprint running horizontal force production capacities at low (FH0) and high (V0) velocities, and HI occurrence in football. This prospective cohort study included 284 football players over one season. All players performed 30 m field sprints at the beginning and different times during the season. Sprint velocity data were used to compute sprint mechanical properties. Players’ injury data were prospectively collected during the entire season. Cox regression analyses were performed using new HI as the outcome, and horizontal force production capacity (FH0 and V0) was used at the start of the season (model 1) and at each measurement time point within the season (model 2) as explanatory variables, adjusted for individual players’ (model 2) age, geographical group of players, height, body mass, and previous HI, with cumulative hours of football practice as the time scale. A total of 47 new HI (20% of all injuries) were observed in 38 out of 284 players (13%). There were no associations between FH0 and/or V0 values at the start of the season and new HI occurrence during the season (model 1). During the season, a total of 801 measurements were performed, from one to six per player. Lower measured FH0 values were significantly associated with a higher risk of sustaining HI within the weeks following sprint measurement (HR = 2.67 (95% CI: 1.51 to 4.73), p < 0.001) (model 2). In conclusion, low horizontal force production capacities at low velocity during early sprint acceleration (FH0) may be considered as a potential additional factor associated with HI risk in a comprehensive, multifactorial, and individualized approach.
Cocaine-Induced Midline Destructive Lesions (CIMDL): A Real Challenge in Diagnosis
Authors: Di Cosola Michele , Ambrosino Mariateresa , Limongelli Luisa , Favia Gianfranco , Santarelli Andrea , Cortelazzi Roberto , Lo Muzio Lorenzo
Published: July 2021

The prolonged use of intranasal cocaine can destroy the nasal architecture with the erosion of the palate, turbinates, and ethmoid sinuses causing cocaine-induced midline lesions (CIMDL). The CIMDL display a clinical pattern mimicking variable diseases. The aim of this study was to highlight the difficulties in reaching a correct diagnosis through the evaluation of eight new cases. The diagnostic procedures followed in these patients included: detailed medical history, clinical and histological examination, computed tomography and magnetic resonance imaging, laboratory findings (complete blood count, sedimentation rate, antinuclear antibody test, rheumatoid factor, venereal disease research laboratory test, leishmaniasis and fungal serology, antineutrophil cytoplasmic antibodies ANCA test), and chest X-ray. All patients complained of epistaxis, halitosis, nasal scabs and obstruction, decreased sense of smell and/or taste, oro-nasal regurgitation of solids and liquids with recurrent sinus infections, and chronic facial pain. On clinical examination, all patients showed palate perforation with variable nasal structure involvement and presented a strong positivity for ANCA tests with a p-ANCA pattern. The followed protocol for the CIMDL diagnosis allowed for a relatively quick and conclusive diagnosis in all patients. A multidisciplinary approach is mandatory in the management of CIMDL, involving dental professionals, maxillofacial surgeons, and psychologists.
Gender Differences in the Extended Theory of Planned Behaviour on Smoking Cessation Intention in Young Soldiers
Authors: Liu Yi-Chun , Yen Li-Chen , Liaw Fang-Yih , Lin Ming-Han , Chiang Shih-Hung , Lin Fu-Gong , Lai Ching-Huang , Kao Senyeong , Chang Yu-Tien , Wu Chia-Chao , Chiu Yu-Lung
Published: July 2021

Background: The theory of planned behaviour (TPB) explanation of smoking cessation intentions consists of gender differences. The purpose of this study is to adopt the extended TPB to discuss factors influencing the smoking cessation intentions of young adult volunteer soldiers and to further compare the respective factors for both genders. Methods: This is a cross-sectional study. Data were collected from 139 and 165 male and female volunteer soldiers who smoked, respectively. Research participants completed a self-administered questionnaire that comprised items pertaining to the participants’ demographic characteristics, smoking behaviours, smoking cessation experiences, social environments, and TPB variables. Results: Subjective norms (friends) are a positive key factor for young adult male (β = 0.033, p = 0.012) and female (β = 0.076, p < 0.001) volunteer soldiers’ smoking cessation intentions, and perceived behavioural control is a key factor for male young (β = 0.226, p = 0.040) adult volunteer soldiers’ smoking cessation intention. The extended TPB accounted for 27.9% and 53.2% of the variance in the intention to quit smoking in the male and female volunteer soldiers, respectively. Conclusions: We suggest that smoking cessation strategies can reinforce gender-specific intervention strategies to assist young adult volunteer soldiers in smoking cessation.
In Silico Structural Modeling and Analysis of Interactions of Tremellomycetes Cytochrome P450 Monooxygenases CYP51s with Substrates and Azoles
Authors: Akapo Olufunmilayo Olukemi , Macnar Joanna M. , Kryś Justyna D. , Syed Puleng Rosinah , Syed Khajamohiddin , Gront Dominik
Published: July 2021

Cytochrome P450 monooxygenase CYP51 (sterol 14α-demethylase) is a well-known target of the azole drug fluconazole for treating cryptococcosis, a life-threatening fungal infection in immune-compromised patients in poor countries. Studies indicate that mutations in CYP51 confer fluconazole resistance on cryptococcal species. Despite the importance of CYP51 in these species, few studies on the structural analysis of CYP51 and its interactions with different azole drugs have been reported. We therefore performed in silico structural analysis of 11 CYP51s from cryptococcal species and other Tremellomycetes. Interactions of 11 CYP51s with nine ligands (three substrates and six azoles) performed by Rosetta docking using 10,000 combinations for each of the CYP51-ligand complex (11 CYP51s × 9 ligands = 99 complexes) and hierarchical agglomerative clustering were used for selecting the complexes. A web application for visualization of CYP51s’ interactions with ligands was developed (http://bioshell.pl/azoledocking/). The study results indicated that Tremellomycetes CYP51s have a high preference for itraconazole, corroborating the in vitro effectiveness of itraconazole compared to fluconazole. Amino acids interacting with different ligands were found to be conserved across CYP51s, indicating that the procedure employed in this study is accurate and can be automated for studying P450-ligand interactions to cater for the growing number of P450s.
Bmi1 Augments Proliferation and Survival of Cortical Bone-Derived Stem Cells after Injury through Novel Epigenetic Signaling via Histone 3 Regulation
Authors: Kraus Lindsay , Bryan Chris , Wagner Marcus , Kino Tabito , Gunchenko Melissa , Jalal Wassy , Khan Mohsin , Mohsin Sadia
Published: July 2021

Ischemic heart disease can lead to myocardial infarction (MI), a major cause of morbidity and mortality worldwide. Multiple stem cell types have been safely transferred into failing human hearts, but the overall clinical cardiovascular benefits have been modest. Therefore, there is a dire need to understand the basic biology of stem cells to enhance therapeutic effects. Bmi1 is part of the polycomb repressive complex 1 (PRC1) that is involved in different processes including proliferation, survival and differentiation of stem cells. We isolated cortical bones stem cells (CBSCs) from bone stroma, and they express significantly high levels of Bmi1 compared to mesenchymal stem cells (MSCs) and cardiac-derived stem cells (CDCs). Using lentiviral transduction, Bmi1 was knocked down in the CBSCs to determine the effect of loss of Bmi1 on proliferation and survival potential with or without Bmi1 in CBSCs. Our data show that with the loss of Bmi1, there is a decrease in CBSC ability to proliferate and survive during stress. This loss of functionality is attributed to changes in histone modification, specifically histone 3 lysine 27 (H3K27). Without the proper epigenetic regulation, due to the loss of the polycomb protein in CBSCs, there is a significant decrease in cell cycle proteins, including Cyclin B, E2F, and WEE as well as an increase in DNA damage genes, including ataxia-telangiectasia mutated (ATM) and ATM and Rad3-related (ATR). In conclusion, in the absence of Bmi1, CBSCs lose their proliferative potential, have increased DNA damage and apoptosis, and more cell cycle arrest due to changes in epigenetic modifications. Consequently, Bmi1 plays a critical role in stem cell proliferation and survival through cell cycle regulation, specifically in the CBSCs. This regulation is associated with the histone modification and regulation of Bmi1, therefore indicating a novel mechanism of Bmi1 and the epigenetic regulation of stem cells.
The Slow-Releasing and Mitochondria-Targeted Hydrogen Sulfide (H2S) Delivery Molecule AP39 Induces Brain Tolerance to Ischemia
Authors: Pomierny Bartosz , Krzyżanowska Weronika , Jurczyk Jakub , Skórkowska Alicja , Strach Beata , Szafarz Małgorzata , Przejczowska-Pomierny Katarzyna , Torregrossa Roberta , Whiteman Matthew , Marcinkowska Monika , Pera Joanna , Budziszewska Bogusława
Published: July 2021

Ischemic stroke is the third leading cause of death in the world, which accounts for almost 12% of the total deaths worldwide. Despite decades of research, the available and effective pharmacotherapy is limited. Some evidence underlines the beneficial properties of hydrogen sulfide (H2S) donors, such as NaSH, in an animal model of brain ischemia and in in vitro research; however, these data are ambiguous. This study was undertaken to verify the neuroprotective activity of AP39, a slow-releasing mitochondria-targeted H2S delivery molecule. We administered AP39 for 7 days prior to ischemia onset, and the potential to induce brain tolerance to ischemia was verified. To do this, we used the rat model of 90-min middle cerebral artery occlusion (MCAO) and used LC-MS/MS, RT-PCR, LuminexTM assays, Western blot and immunofluorescent double-staining to determine the absolute H2S levels, inflammatory markers, neurotrophic factor signaling pathways and apoptosis marker in the ipsilateral frontal cortex, hippocampus and in the dorsal striatum 24 h after ischemia onset. AP39 (50 nmol/kg) reduced the infarct volume, neurological deficit and reduced the microglia marker (Iba1) expression. AP39 also exerted prominent anti-inflammatory activity in reducing the release of Il-1β, Il-6 and TNFα in brain areas particularly affected by ischemia. Furthermore, AP39 enhanced the pro-survival pathways of neurotrophic factors BDNF-TrkB and NGF-TrkA and reduced the proapoptotic proNGF-p75NTR-sortilin pathway activity. These changes corresponded with reduced levels of cleaved caspase 3. Altogether, AP39 treatment induced adaptative changes within the brain and, by that, developed brain tolerance to ischemia.
TSA Promotes CRISPR/Cas9 Editing Efficiency and Expression of Cell Division-Related Genes from Plant Protoplasts
Authors: Choi Seung Hee , Lee Myoung Hui , Jin Da Mon , Ju Su Ji , Ahn Woo Seok , Jie Eun Yee , Lee Ji Min , Lee Jiyoung , Kim Cha Young , Kim Suk Weon
Published: July 2021

Trichostatin A (TSA) is a representative histone deacetylase (HDAC) inhibitor that modulates epigenetic gene expression by regulation of chromatin remodeling in cells. To investigate whether the regulation of chromatin de-condensation by TSA can affect the increase in the efficiency of Cas9 protein-gRNA ribonucleoprotein (RNP) indel formation from plant cells, genome editing efficiency using lettuce and tobacco protoplasts was examined after several concentrations of TSA treatments (0, 0.1, 1 and 10 μM). RNP delivery from protoplasts was conducted by conventional polyethylene glycol (PEG) transfection protocols. Interestingly, the indel frequency of the SOC1 gene from TSA treatments was about 3.3 to 3.8 times higher than DMSO treatment in lettuce protoplasts. The TSA-mediated increase of indel frequency of the SOC1 gene in lettuce protoplasts occurred in a concentration-dependent manner, although there was not much difference. Similar to lettuce, TSA also increased the indel frequency by 1.5 to 1.8 times in a concentration-dependent manner during PDS genome editing using tobacco protoplasts. The MNase test clearly showed that chromatin accessibility with TSA treatments was higher than that of DMSO treatment. Additionally, TSA treatment significantly increased the level of histone H3 and H4 acetylation from lettuce protoplasts. The qRT-PCR analysis showed that expression of cell division-related genes (LsCYCD1-1, LsCYCD3-2, LsCYCD6-1, and LsCYCU4-1) was increased by TSA treatment. These findings could contribute to increasing the efficiency of CRISPR/Cas9-mediated genome editing. Furthermore, this could be applied for the development of useful genome-edited crops using the CRISPR/Cas9 system with plant protoplasts.
Trp Fluorescence Redshift during HDL Apolipoprotein Denaturation Is Increased in Patients with Coronary Syndrome in Acute Phase: A New Assay to Evaluate HDL Stability
Authors: López-Olmos Victoria , Luna-Luna María , Carreón-Torres Elizabeth , González-Pacheco Héctor , Bautista-Pérez Rocío , Posadas-Sánchez Rosalinda , Fragoso José Manuel , Vargas-Alarcón Gilberto , Pérez-Méndez Óscar
Published: July 2021

High-density lipoproteins’ (HDL) stability is a determinant of their residence times in plasma and consequently an important parameter that influences the beneficial properties of these lipoproteins. Since there are no accessible procedures for this purpose, here, we describe the methodological conditions to assess the stability of the HDL based on the redshift of the fluorescence spectrum of tryptophans contained in the structure of HDL-apolipoproteins during incubation with urea 8M. Along the HDL denaturation kinetics, the main variations of fluorescence were observed at the wavelengths of 330, 344, and 365 nm at room temperature. Therefore, HDL denaturation was estimated using the tryptophan (Trp)-ratio of fluorescence intensity (rfi) at such wavelengths. By setting 100% of the measurable denaturation at 26 h, HDL reached 50% after 8 h of incubation with urea. Then, for further analyses we determined the percentage of HDL denaturation at 8 h as an estimation of the stability of these lipoproteins. To explore the potential usefulness of this test, we analyzed the stability of HDL isolated from the plasma of 24 patients diagnosed with acute coronary syndrome (ACS). These HDL presented significantly higher percentages of denaturation (64.9% (58.7–78.4)) than HDLs of healthy individuals (23.3% (20.3-27.0)). These results indicate that HDL in ACS are less stable than in control subjects. Moreover, the percentage of denaturation of HDL correlated with body mass index and aspartate transaminase plasma activity. Furthermore, apo-I, HDL-cholesterol, HDL-triglycerides, and apo A-I-to-triglycerides ratio correlated with the percentage of HDL denaturation, suggesting that the lipoprotein composition is a main determinant of HDL stability. Finally, the percentage of HDL denaturation is the parameter that predicted the presence of ACS as determined by a machine learning procedure and logistic regression analysis. In conclusion, we established the methodological conditions to assess the stability of HDL by a fluorescence-based method that merits exploration in prospective studies for evaluating the coronary artery disease risk.
Interaction between Angiotensinase Activities in Pituitary and Adrenal Glands of Wistar–Kyoto and Spontaneously Hypertensive Rats under Hypotensive or Hypertensive Treatments
Authors: Segarra Ana B. , Prieto Isabel , Banegas Inmaculada , Martínez-Cañamero Magdalena , Villarejo Ana B. , Domínguez-Vías Germán , de Gasparo Marc , Ramírez-Sánchez Manuel
Published: July 2021

In the present study, we analyzed the activity of several aminopeptidases (angiotensinases) involved in the metabolism of various angiotensin peptides, in pituitary and adrenal glands of untreated Wistar–Kyoto (WKY) and spontaneously hypertensive rats (SHR) or treated with the antihypertensive drugs captopril and propranolol or with the L-Arginine hypertensive analogue L-NG-Nitroarginine Methyl Ester (L-NAME). Intra- and inter-gland correlations between angiotensinase activities were also calculated. Membrane-bound alanyl-, cystinyl-, and glutamyl-aminopeptidase activities were determined fluorometrically using aminoacyl-β-naphthylamide as substrates. Depending on the type of angiotensinase analyzed, the results reflect a complex picture showing substantial differences between glands, strains, and treatments. Alanyl-aminopeptidase responsible for the metabolism of Ang III to Ang IV appears to be the most active angiotensinase in both pituitary and adrenals of WKY and particularly in SHR. Independently of treatment, most positive correlations are observed in the pituitary gland of WKY whereas such positive correlations are predominant in adrenals of SHR. Negative inter-gland correlations were observed in control SHR and L-NAME treated WKY. Positive inter-gland correlations were observed in captopril-treated SHR and propranolol-treated WKY. These results may reflect additional mechanisms for increasing or decreasing systolic blood pressure in WKY or SHR.
Microglia and the Aging Brain: Are Geriatric Microglia Linked to Poor Sleep Quality?
Authors: Choudhury Mohammed E. , Miyanishi Kazuya , Takeda Haruna , Tanaka Junya
Published: July 2021

Poor sleep quality and disrupted circadian behavior are a normal part of aging and include excessive daytime sleepiness, increased sleep fragmentation, and decreased total sleep time and sleep quality. Although the neuronal decline underlying the cellular mechanism of poor sleep has been extensively investigated, brain function is not fully dependent on neurons. A recent antemortem autographic study and postmortem RNA sequencing and immunohistochemical studies on aged human brain have investigated the relationship between sleep fragmentation and activation of the innate immune cells of the brain, microglia. In the process of aging, there are marked reductions in the number of brain microglial cells, and the depletion of microglial cells disrupts circadian rhythmicity of brain tissue. We also showed, in a previous study, that pharmacological suppression of microglial function induced sleep abnormalities. However, the mechanism underlying the contribution of microglial cells to sleep homeostasis is only beginning to be understood. This review revisits the impact of aging on the microglial population and activation, as well as microglial contribution to sleep maintenance and response to sleep loss. Most importantly, this review will answer questions such as whether there is any link between senescent microglia and age-related poor quality sleep and how this exacerbates neurodegenerative disease.
Angulin-1 (LSR) Affects Paracellular Water Transport, However Only in Tight Epithelial Cells
Authors: Ayala-Torres Carlos , Krug Susanne M. , Rosenthal Rita , Fromm Michael
Published: July 2021

Water transport in epithelia occurs transcellularly (aquaporins) and paracellularly (claudin-2, claudin-15). Recently, we showed that downregulated tricellulin, a protein of the tricellular tight junction (tTJ, the site where three epithelial cells meet), increased transepithelial water flux. We now check the hypothesis that another tTJ-associated protein, angulin-1 (alias lipolysis-stimulated lipoprotein receptor, LSR) is a direct negative actuator of tTJ water permeability depending on the tightness of the epithelium. For this, a tight and an intermediate-tight epithelial cell line, MDCK C7 and HT-29/B6, were stably transfected with CRISPR/Cas9 and single-guide RNA targeting angulin-1 and morphologically and functionally characterized. Water flux induced by an osmotic gradient using 4-kDa dextran caused water flux to increase in angulin-1 KO clones in MDCK C7 cells, but not in HT-29/B6 cells. In addition, we found that water permeability in HT-29/B6 cells was not modified after either angulin-1 knockout or tricellulin knockdown, which may be related to the presence of other pathways, which reduce the impact of the tTJ pathway. In conclusion, modulation of the tTJ by knockout or knockdown of tTJ proteins affects ion and macromolecule permeability in tight and intermediate-tight epithelial cell lines, while the transepithelial water permeability was affected only in tight cell lines.
Differential Domain Distribution of gnomAD- and Disease-Linked Connexin Missense Variants
Authors: Bai Donglin , Wang Jiayi , Li Tianhe , Chan Ryan , Atalla Mena , Chen Robert C. , Khazaneh Mohammad T. , An Raphael J. , Stathopulos Peter B.
Published: July 2021

Twenty-one human genes encode connexins, a family of homologous proteins making gap junction (GJ) channels, which mediate direct intercellular communication to synchronize tissue/organ activities. Genetic variants in more than half of the connexin genes are associated with dozens of different Mendelian inherited diseases. With rapid advances in DNA sequencing technology, more variants are being identified not only in families and individuals with diseases but also in people in the general population without any apparent linkage to Mendelian inherited diseases. Nevertheless, it remains challenging to classify the pathogenicity of a newly identified connexin variant. Here, we analyzed the disease- and Genome Aggregation Database (gnomAD, as a proxy of the general population)-linked variants in the coding region of the four disease-linked α connexin genes. We found that the most abundant and position-sensitive missense variants showed distinct domain distribution preference between disease- and gnomAD-linked variants. Plotting missense variants on topological and structural models revealed that disease-linked missense variants are highly enriched on the structurally stable/resolved domains, especially the pore-lining domains, while the gnomAD-linked missense variants are highly enriched in the structurally unstable/unresolved domains, especially the carboxyl terminus. In addition, disease-linked variants tend to be on highly conserved residues and those positions show evolutionary co-variation, while the gnomAD-linked missense variants are likely on less conserved residue positions and on positions without co-variation. Collectively, the revealed distribution patterns of disease- and gnomAD-linked missense variants further our understanding of the GJ structure–biological function relationship, which is valuable for classifying the pathogenicity of newly identified connexin variants.
Definition of the Binding Architecture to a Target Promoter of HP1043, the Essential Master Regulator of Helicobacter pylori
Authors: Zannoni Annamaria , Pelliciari Simone , Musiani Francesco , Chiappori Federica , Roncarati Davide , Scarlato Vincenzo
Published: July 2021

HP1043 is an essential orphan response regulator of Helicobacter pylori orchestrating multiple crucial cellular processes. Classified as a member of the OmpR/PhoB family of two-component systems, HP1043 exhibits a highly degenerate receiver domain and evolved to function independently of phosphorylation. Here, we investigated the HP1043 binding mode to a target sequence in the hp1227 promoter (Php1227). Scanning mutagenesis of HP1043 DNA-binding domain and consensus sequence led to the identification of residues relevant for the interaction of the protein with a target DNA. These determinants were used as restraints to guide a data-driven protein-DNA docking. Results suggested that, differently from most other response regulators of the same family, HP1043 binds in a head-to-head conformation to the Php1227 target promoter. HP1043 interacts with DNA largely through charged residues and contacts with both major and minor grooves of the DNA are required for a stable binding. Computational alanine scanning on molecular dynamics trajectory was performed to corroborate our findings. Additionally, in vitro transcription assays confirmed that HP1043 positively stimulates the activity of RNA polymerase.
Mesenchymal Stem Cell-Based Therapy as an Alternative to the Treatment of Acute Respiratory Distress Syndrome: Current Evidence and Future Perspectives
Authors: Fernández-Francos Silvia , Eiro Noemi , González-Galiano Natalia , Vizoso Francisco J.
Published: July 2021

Acute respiratory distress syndrome (ARDS) represents a current challenge for medicine due to its incidence, morbidity and mortality and, also, the absence of an optimal treatment. The COVID-19 outbreak only increased the urgent demand for an affordable, safe and effective treatment for this process. Early clinical trials suggest the therapeutic usefulness of mesenchymal stem cells (MSCs) in acute lung injury (ALI) and ARDS. MSC-based therapies show antimicrobial, anti-inflammatory, regenerative, angiogenic, antifibrotic, anti-oxidative stress and anti-apoptotic actions, which can thwart the physiopathological mechanisms engaged in ARDS. In addition, MSC secretome and their derived products, especially exosomes, may reproduce the therapeutic effects of MSC in lung injury. This last strategy of treatment could avoid several safety issues potentially associated with the transplantation of living and proliferative cell populations and may be formulated in different forms. However, the following diverse limitations must be addressed: (i) selection of the optimal MSC, bearing in mind both the heterogeneity among donors and across different histological origins, (ii) massive obtention of these biological products through genetic manipulations of the most appropriate MSC, (iii) bioreactors that allow their growth in 3D, (iv) ideal culture conditions and (v) adequate functional testing of these obtaining biological products before their clinical application.
Skin Mast Cell-Driven Ceramides Drive Early Apoptosis in Pre-Symptomatic Eczema in Mice
Authors: Robida Piper A. , Chumanevich Alena P. , Gandy Alexa Orr , Fuseler John W. , Nagarkatti Prakash , Nagarkatti Mitzi , Oskeritzian Carole A.
Published: July 2021

Atopic dermatitis (AD or eczema) is the most common chronic inflammatory skin disorder worldwide. Ceramides (Cer) maintain skin barrier functions, which are disrupted in lesional skin of AD patients. However, Cer status during the pre-lesional phase of AD is not well defined. Using a variation of human AD-like preclinical model consisting of a 7-day topical exposure to ovalbumin (OVA), or control, we observed elevation of Cer C16 and C24. Skin mRNA quantification of enzymes involved in Cer metabolism [Cer synthases (CerS) and ceramidases (Asah1/Asah2)], which revealed augmented CerS 4, 5 and 6 and Asah1. Given the overall pro-apoptotic nature of Cer, local apoptosis was assessed, then quantified using novel morphometric measurements of cleaved caspase (Casp)-3-restricted immunofluorescence signal in skin samples. Apoptosis was induced in response to OVA. Because apoptosis may occur downstream of endoplasmic reticulum (ER) stress, we measured markers of ER stress-induced apoptosis and found elevated skin-associated CHOP protein upon OVA treatment. We previously substantiated the importance of mast cells (MC) in initiating early skin inflammation. OVA-induced Cer increase and local apoptosis were prevented in MC-deficient mice; however, they were restored following MC reconstitution. We propose that the MC/Cer axis is an essential pathogenic feature of pre-lesional AD, whose targeting may prevent disease development.
Chronic Prostatitis/Chronic Pelvic Pain Syndrome Leads to Impaired Semen Parameters, Increased Sperm DNA Fragmentation and Unfavorable Changes of Sperm Protamine mRNA Ratio
Authors: Berg Elena , Houska Petr , Nesheim Nils , Schuppe Hans-Christian , Pilatz Adrian , Fijak Monika , Manthey Marc , Steger Klaus , Wagenlehner Florian , Schagdarsurengin Undraga
Published: July 2021

Background: Chronic Prostatitis/Chronic Pelvic Pain Syndrome (CP/CPPS) is a frequent disease affecting men of every age and accounting for a great number of consultations at urology departments. Previous studies suggested a negative impact of CP/CPPS on fertility. As increasing attention has been attributed to additional aspects, such as sperm DNA integrity and sperm protein alterations, besides the WHO standard semen analysis when assessing male fertility, in this prospective study, we aimed to further characterize the fertility status in CP/CPPS patients with a focus on these parameters. Methods: Sperm DNA fragmentation measured by sperm chromatin structure assay (SCSA) and protamine 1 to protamine 2 mRNA ratio assessed by RT-qPCR were analyzed along with conventional ejaculate parameters and inflammatory markers in 41 CP/CPPS patients and 22 healthy volunteers. Results: We found significant differences between the groups concerning multiple conventional ejaculate parameters. A significant increase in sperm DNA fragmentation was shown in CP/CPPS patients with association to other sperm parameters. The majority of CP/CPPS patients exhibited protamine mRNA ratios out of the range of regular fertility. Conclusions: This is a pioneering study with a strong practical orientation revealing that CP/CPPS leads to increased sperm DNA damage and changes in sperm protamine levels, emphasizing an unfavorable impact of CP/CPPS on fertility.
Effect of Kinesin-5 Tail Domain on Motor Dynamics for Antiparallel Microtubule Sliding
Authors: Liu Yuying , Wang Yao , Wang Pengye , Xie Ping
Published: July 2021

Kinesin-5 motor consists of two pairs of heads and tail domains, which are situated at the opposite ends of a common stalk. The two pairs of heads can bind to two antiparallel microtubules (MTs) and move on the two MTs independently towards the plus ends, sliding apart the two MTs, which is responsible for chromosome segregation during mitosis. Prior experimental data showed that the tails of kinesin-5 Eg5 can modulate the dynamics of single motors and are critical for multiple motors to generate high steady forces to slide apart two antiparallel MTs. To understand the molecular mechanism of the tails modulating the ability of Eg5 motors, based on our proposed model the dynamics of the single Eg5 with the tails and that without the tails moving on single MTs is studied analytically and compared. Furthermore, the dynamics of antiparallel MT sliding by multiple Eg5 motors with the tails and that without the tails is studied numerically and compared. Both the analytical results for single motors and the numerical results for multiple motors are consistent with the available experimental data.
Reversing the Psychiatric Effects of Neurodevelopmental Cannabinoid Exposure: Exploring Pharmacotherapeutic Interventions for Symptom Improvement
Authors: De Felice Marta , Laviolette Steven R.
Published: July 2021

Neurodevelopmental exposure to psychoactive compounds in cannabis, specifically THC, is associated with a variety of long-term psychopathological outcomes. This increased risk includes a higher prevalence of schizophrenia, mood and anxiety disorders, and cognitive impairments. Clinical and pre-clinical research continues to identify a wide array of underlying neuropathophysiological sequelae and mechanisms that may underlie THC-related psychiatric risk vulnerability, particularly following adolescent cannabis exposure. A common theme among these studies is the ability of developmental THC exposure to induce long-term adaptations in the mesocorticolimbic system which resemble pathological endophenotypes associated with these disorders. This narrative review will summarize recent clinical and pre-clinical evidence that has elucidated these THC-induced developmental risk factors and examine how specific pharmacotherapeutic interventions may serve to reverse or perhaps prevent these cannabis-related risk outcomes.
Delayed Double Treatment with Adult-Sourced Adipose-Derived Mesenchymal Stem Cells Increases Striatal Medium-Spiny Neuronal Number, Decreases Striatal Microglial Number, and Has No Subventricular Proliferative Effect, after Acute Neonatal Hypoxia-Ischemia in Male Rats
Authors: Basham Haylee K. , Aghoghovwia Benjamin E. , Papaioannou Panagiotis , Seo Steve , Oorschot Dorothy E.
Published: July 2021

Perinatal hypoxia-ischemia (HI) is a major cause of striatal injury. Delayed post-treatment with adult-sourced bone marrow-derived mesenchymal stem cells (BMSCs) increased the absolute number of striatal medium-spiny neurons (MSNs) following perinatal HI-induced brain injury. Yet extraction of BMSCs is more invasive and difficult compared to extraction of adipose-derived mesenchymal stem cells (AD-MSCs), which are easily sourced from subcutaneous tissue. Adult-sourced AD-MSCs are also superior to BMSCs in the treatment of adult ischemic stroke. Therefore, we investigated whether delayed post-treatment with adult-sourced AD-MSCs increased the absolute number of striatal MSNs following perinatal HI-induced brain injury. This included investigation of the location of injected AD-MSCs within the brain, which were widespread in the dorsolateral subventricular zone (dlSVZ) at 1 day after their injection. Cells extracted from adult rat tissue were verified to be stem cells by their adherence to tissue culture plastic and their expression of specific ‘cluster of differentiation’ (CD) markers. They were verified to be AD-MSCs by their ability to differentiate into adipocytes and osteocytes in vitro. Postnatal day (PN) 7/8, male Sprague-Dawley rats were exposed to either HI right-sided brain injury or no HI injury. The HI rats were either untreated (HI + Diluent), single stem cell-treated (HI + MSCs×1), or double stem cell-treated (HI + MSCs×2). Control rats that were matched-for-weight and litter had no HI injury and were treated with diluent (Uninjured + Diluent). Treatment with AD-MSCs or diluent occurred either 7 days, or 7 and 9 days, after HI. There was a significant increase in the absolute number of striatal dopamine and cyclic AMP-regulated phosphoprotein (DARPP-32)-positive MSNs in the double stem cell-treated (HI + MSCs×2) group and the normal control group compared to the HI + Diluent group at PN21. We therefore investigated two potential mechanisms for this effect of double-treatment with AD-MSCs. Specifically, did AD-MSCs: (i) increase the proliferation of cells within the dlSVZ, and (ii) decrease the microglial response in the dlSVZ and striatum? It was found that a primary repair mechanism triggered by double treatment with AD-MSCs involved significantly decreased striatal inflammation. The results may lead to the development of clinically effective and less invasive stem cell therapies for neonatal HI brain injury.
Critical Roles of Lysophospholipid Receptors in Activation of Neuroglia and Their Neuroinflammatory Responses
Authors: Gaire Bhakta Prasad , Choi Ji-Woong
Published: July 2021

Activation of microglia and/or astrocytes often releases proinflammatory molecules as critical pathogenic mediators that can promote neuroinflammation and secondary brain damages in diverse diseases of the central nervous system (CNS). Therefore, controlling the activation of glial cells and their neuroinflammatory responses has been considered as a potential therapeutic strategy for treating neuroinflammatory diseases. Recently, receptor-mediated lysophospholipid signaling, sphingosine 1-phosphate (S1P) receptor- and lysophosphatidic acid (LPA) receptor-mediated signaling in particular, has drawn scientific interest because of its critical roles in pathogenies of diverse neurological diseases such as neuropathic pain, systemic sclerosis, spinal cord injury, multiple sclerosis, cerebral ischemia, traumatic brain injury, hypoxia, hydrocephalus, and neuropsychiatric disorders. Activation of microglia and/or astrocytes is a common pathogenic event shared by most of these CNS disorders, indicating that lysophospholipid receptors could influence glial activation. In fact, many studies have reported that several S1P and LPA receptors can influence glial activation during the pathogenesis of cerebral ischemia and multiple sclerosis. This review aims to provide a comprehensive framework about the roles of S1P and LPA receptors in the activation of microglia and/or astrocytes and their neuroinflammatory responses in CNS diseases.
Macroalgal Defense against Competitors and Herbivores
Authors: Budzałek Gracjana , Śliwińska-Wilczewska Sylwia , Wiśniewska Kinga , Wochna Agnieszka , Bubak Iwona , Latała Adam , Wiktor Józef Maria
Published: July 2021

Macroalgae are the source of many harmful allelopathic compounds, which are synthesized as a defense strategy against competitors and herbivores. Therefore, it can be predicted that certain species reduce aquaculture performance. Herein, the allelopathic ability of 123 different taxa of green, red, and brown algae have been summarized based on literature reports. Research on macroalgae and their allelopathic effects on other animal organisms was conducted primarily in Australia, Mexico, and the United States. Nevertheless, there are also several scientific reports in this field from South America and Asia; the study areas in the latter continents coincide with areas where aquaculture is highly developed and widely practiced. Therefore, the allelopathic activity of macroalgae on coexisting animals is an issue that is worth careful investigation. In this work, we characterize the distribution of allelopathic macroalgae and compare them with aquaculture locations, describe the methods for the study of macroalgal allelopathy, present the taxonomic position of allelopathic macroalgae and their impact on coexisting aquatic competitors (Cnidaria) and herbivores (Annelida, Echinodermata, Arthropoda, Mollusca, and Chordata), and compile information on allelopathic compounds produced by different macroalgae species. This work gathers the current knowledge on the phenomenon of macroalgal allelopathy and their allelochemicals affecting aquatic animal (competitors and predators) worldwide and it provides future research directions for this topic.
Current Approaches for Advancement in Understanding the Molecular Mechanisms of Mycotoxin Biosynthesis
Authors: Gallo Antonia , Perrone Giancarlo
Published: July 2021

Filamentous fungi are able to synthesise a remarkable range of secondary metabolites, which play various key roles in the interaction between fungi and the rest of the biosphere, determining their ecological fitness. Many of them can have a beneficial activity to be exploited, as well as negative impact on human and animal health, as in the case of mycotoxins contaminating large quantities of food, feed, and agricultural products worldwide and posing serious health and economic risks. The elucidation of the molecular aspects of mycotoxin biosynthesis has been greatly sped up over the past decade due to the advent of next-generation sequencing technologies, which greatly reduced the cost of genome sequencing and related omic analyses. Here, we briefly highlight the recent progress in the use and integration of omic approaches for the study of mycotoxins biosynthesis. Particular attention has been paid to genomics and transcriptomic approaches for the identification and characterisation of biosynthetic gene clusters of mycotoxins and the understanding of the regulatory pathways activated in response to physiological and environmental factors leading to their production. The latest innovations in genome-editing technology have also provided a more powerful tool for the complete explanation of regulatory and biosynthesis pathways. Finally, we address the crucial issue of the interpretation of the combined omics data on the biology of the mycotoxigenic fungi. They are rapidly expanding and require the development of resources for more efficient integration, as well as the completeness and the availability of intertwined data for the research community.