bluuebunny/crossref_metadata_2025 huggingface dataset:
A Statistical Analysis of Death Rates in Italy for the Years 2015–2020 and a Comparison with the Casualties Reported from the COVID-19 Pandemic
Authors: Bonifazi Gianluca , Lista Luca , Menasce Dario , Mezzetto Mauro , Oliva Alberto , Pedrini Daniele , Spighi Roberto , Zoccoli Antonio
Published: April 2021

We analyze the data about casualties in Italy in the period 1 January 2015 to 30 September 2020 released by the Italian National Institute of Statistics (ISTAT). The aim of this article was the description of a statistically robust methodology to extract quantitative values for the seasonal excesses of deaths featured by the data, accompanying them with correct estimates of the relative uncertainties. We will describe the advantages of the method adopted with respect to others listed in literature. The data exhibit a clear sinusoidal behavior, whose fit allows for a robust subtraction of the baseline trend of casualties in Italy, with a surplus of mortality in correspondence to the flu epidemics in winter and to the hottest periods in summer. The overall quality of the fit to the data turns out to be very good, an indication of the validity of the chosen model. We discuss the trend of casualties in Italy by different classes of ages and for the different genders. We finally compare the data-subtracted casualties, as reported by ISTAT, with those reported by the Italian Department for Civil Protection (DPC) relative to the deaths directly attributed to the Coronavirus Disease 2019 caused by the SARS-CoV-2 virus (COVID-19), and we point out the differences in the two samples, collected under different assumptions.
The Applicability of the Poincaré Plot in the Analysis of Variability of Reaction Time during Serial Testing
Authors: Iconaru Elena Ioana , Ciucurel Manuela Mihaela , Georgescu Luminita , Tudor Mariana , Ciucurel Constantin
Published: April 2021

(1) Background: This study aims to put into evince the relationship between the variability of the reaction time (RT) during repeated testing, expressed through indicators extracted by the Poincaré plot method, and the age of the participants, their self-reported health (SRH), and level of perceived anxiety. (2) Methods: The study was performed using computerized RT testing software. An observational cross-sectional study was performed on a group of 120 subjects (mean age 42.33 ± 21.12 years), sex ratio men to women 1.14:1. Data were processed through descriptive and inferential statistics. The Poincaré plot method was applied in the analysis of the RT series of data, by calculating the indicators SD1, SD2, SD1/SD2, and area of the fitting ellipse (AFE) (3) Results: We provided evidence of the excellent reliability of the web-based RT serial testing (Cronbach’s Alpha 0.991) with this sample group. Our results showed that age is an important predictor for mean values of RT, while SD1, SD2, and AFE indicators are for SRH (p < 0.01). (4) Conclusions: the variability of RT, expressed by the Poincaré plot indicators, reflects the health status rather than the aging of the subjects and is barely influenced by their level of anxiety.
SARS-CoV-2 Infection among the Dental Staff from Lombardy Region, Italy
Authors: Gallus Silvano , Paroni Luca , Re Dino , Aiuto Riccardo , Battaglia Davide Maria , Crippa Rolando , Carugo Nicolò , Beretta Matteo , Balsano Lorenzo , Paglia Luigi
Published: April 2021

Dentists have been supposed to be among the healthcare workers at greatest risk of SARS-CoV-2 infection. However, scant data are available on the issue. The aim of this study is to quantify the SARS-CoV-2 antibody prevalence and determinants in a sample of dentists, dental hygienists, and other personnel employed among the dental staff in Lombardy region. We used an accurate rapid diagnostic test kit detecting immunoglobulins (Ig) in 504 adults. Of the 499 participants who obtained a valid antibody test, 54 (10.8%) had a SARS-CoV-2 positive test (0.4% IgM+, 1.8% both IgM+ and IgG+, and 8.6% IgG+). A statistically significant association with infection was found for geographic area (compared to Milan, adjusted odds ratio was 2.79, 95% confidence interval, CI: 1.01–7.68 for eastern and 2.82, 95% CI: 1.34–5.94, for southern Lombardy). The clinical staff did not result positive to SARS-CoV-2 more frequently than the administrative staff. This is the first study using antibody test in the dental staff personnel. It shows that the prevalence of SARS-CoV-2 infection in Lombardy region was around 10%, in line with estimates on other healthcare professionals. Despite the close physical contact with the patient, dentists have been able to scrupulously manage and effectively use protective devices.
The Association between Gait Speed and Falls in Community Dwelling Older Adults with and without Mild Cognitive Impairment
Authors: Adam Claire E. , Fitzpatrick Annette L. , Leary Cindy S. , Hajat Anjum , Phelan Elizabeth A. , Park Christina , Semmens Erin O.
Published: April 2021

(1) Background: Falls are common in older adults and result in injuries, loss of independence, and death. Slow gait is associated with falls in older adults, but few studies have assessed the association between gait speed and falls among those with mild cognitive impairment (MCI). (2) Methods: The association between gait speed and falls was assessed in 2705 older adults with and without MCI participating in the Ginkgo Evaluation of Memory Study. Gait speed was measured via a 15-foot walk test and fall history through self-report. We used data collected at the 12-month (2001–2003) and 18-month visits (2002–2004). (3) Results: Participant average age was 78.5 years (sd = 3.2); 45% were female, and 14% had MCI at baseline. The average gait speed was 0.93 m/s (sd = 0.20). Sixteen percent (n = 433) and 18% (n = 498) reported at least one fall at the 12-month and 18-month visits, respectively. Faster gait speed was associated with decreased risk of falling (RR: 0.95, 95% CI: 0.91, 0.99) for every 10 cm/s increase in gait speed adjusted for age, gender, study arm, site, and MCI status. (4) Conclusions: The relationship between gait speed and risk of falling did not vary by MCI status (interaction p-value = 0.78).
Nurses’ Perceptions on the Implementation of a Safe Drug Administration Protocol and Its Effect on Error Notification
Authors: Escandell-Rico Francisco Miguel , Perpiñá-Galvañ Juana , Pérez-Fernández Lucía , Sanjuán-Quiles Ángela , Gómez-Beltrán Piedras Albas , Ramos-Pichardo Juan Diego
Published: April 2021

Patient safety and quality of care are fundamental pillars in the health policies of various governments and international organizations. The purpose of this study is to evaluate nurses’ perceptions on the degree of implementation of a protocol for the standardization of care and to measure its influence on notification of adverse events related to the administration of medications. This comparative study used data obtained from questionnaires completed by 180 nurses from medical and surgical units. Our analyses included analysis of variance and regression models. We observe that the responses changed unevenly over time in each group, finding significant differences in all comparisons. The mean response rating was increased at 6 months in the intervention group, and this level was maintained at 12 months. With the new protocol, a total of 246 adverse events and 481 incidents without harm was reported. Thus, actions such as the use of protocols and event notification systems should be implemented to improve quality of care and patient safety.
Effects of Mirror Therapy Combined with EMG-Triggered Functional Electrical Stimulation to Improve on Standing Balance and Gait Ability in Patient with Chronic Stroke
Authors: Kim Dong-Hoon , Jang Sang-Hun
Published: April 2021

This study was performed to evaluate the effects of EMG-triggered functional electrical stimulation on balance and gait ability on patient with Chronic Stroke. A total of 60 chronic stroke patients were divided into mirror treatment and functional electrical (MT-EF) Group, MT group, CON group. Each group performed 60 min a day five times a week for eight weeks. MT-FE group was performed 30 min five times a week for eight weeks in mirror therapy process with EMG-FES. MT group performed 30 min five times a week for eight weeks in mirror therapy process. CON group was performed 30 min five times a week for eight weeks in conservative treatment. To measure the balance ability, Biorescue (COP, LOS), Berg balance scale (BBS) and FRT, and the gait ability test was performed by 10 m walk test. MT-FE group revealed significant differences in COP, LOS, BBS, FRT and 10 m walk test as compared to the MT and CON groups (p < 0.05). Our results showed that MT-FE was more effective on COP, LOS, BBS, FRT and 10 m walk test in patients with chronic stroke. Our results also showed that MT-EF group was more effective on balance and gait ability in patients with chronic stroke. We suggest that this study can be used for intervention data for recovering balance and gait ability in chronic stroke patients.
Burnout and Engagement Dimensions in the Reception System of Illegal Immigration in the Mediterranean Sea. A Qualitative Study on a Sample of Italian Practitioners
Authors: Nonnis Marcello , Agus Mirian , Pirrone Monica Piera , Cuccu Stefania , Pedditzi Maria Luisa , Cortese Claudio Giovanni
Published: April 2021

The present study describes the semantic nature of burnout and engagement in the operators involved in the management of illegal immigration. Semi-structured interviews were conducted on a sample of Italian practitioners (n = 62) of the two levels of the reception system considered: (1) rescue and first aid and (2) reception and integration. Within the framework of the job demands–resources model (JD-R), the interviews deepened the analysis of the positive and negative dimensions of burnout and engagement: exhaustion versus energy, relational deterioration versus relational involvement, professional inefficacy versus professional efficacy and disillusion versus trust. The interviews were analysed using the T-Lab software, through a cluster analysis (bisecting K-means algorithm), which emphasised noteworthy themes. The results show that, in the vast majority of the dimensions considered (for both levels of reception), the same dimensions of engagement of the operators (energy, relational involvement, professional efficacy and trust) are able to lead them into a condition of burnout, with experiences, conversely, of exhaustion, relational deterioration, professional inefficacy and disillusion. These findings expand the knowledge on burnout and engagement in practitioners of illegal immigration, a context characterised by the value of help and welcome.
Occurrence, Potential Sources, and Risk Assessment of Volatile Organic Compounds in the Han River Basin, South Korea
Authors: Im Jong Kwon , Yu Soon Ju , Kim Sujin , Kim Sang Hun , Noh Hye Ran , Kim Moon Kyung
Published: April 2021

Increasing public awareness about the aesthetics and safety of water sources has shifted researchers’ attention to the adverse effects of volatile organic compounds (VOCs) on humans and aquatic organisms. A total of 17 VOCs, including 10 volatile halogenated hydrocarbons and seven volatile non-halogenated hydrocarbons, were investigated at 36 sites of the Han River Basin, which is the largest and most important drinking water source for residents of the Seoul metropolitan area and Gyeonggi province in South Korea. The VOC concentrations ranged from below detection limits to 1.813 µg L−1. The most frequently detected VOC was 1,2-dichloropropane, with a detection frequency of 80.56%, as it is used as a soil fumigant, chemical intermediate, and industrial solvent. In terms of geographical trends, the sampling sites that were under the influence of sewage and industrial wastewater treatment plants were more polluted with VOCs than other areas. This observation was also supported by the results of the principal component analysis. In the present study, the detected concentrations of VOCs were much lower than that of the predicted no-effect concentrations, suggesting low ecological risk in the Han River. However, a lack of available ecotoxicity data and limited comparable studies warrants further studies on these compounds.
Risk Factors for Acute Urticaria in Central California
Authors: Jadhav Rohan , Alcala Emanuel , Sirota Sarah , Capitman John
Published: April 2021

At least 15–20% of the population in the world suffers from urticaria. Allergy triggers contribute to the development of urticaria. Not much is known about the demographic and environmental risk factors that contribute to the occurrence of acute urticaria. Methods: We utilized emergency department data on acute urticaria-related visits managed by the California Office of Statewide Planning and Operations for 201 zip codes located in southern central California (San Joaquin Valley) collected during the years 2016 and 2017. Census data from the same zip codes were considered as a population at risk. Socioeconomic and environmental parameters using CalEnviroScreen (Office of Environmental Health Hazard Assessment, Sacramento, CA, USA) database for the zip codes were evaluated as risk factors. Results: The incidence rate of acute urticaria in San Joaquin Valley during 2016–2017 was 1.56/1000 persons (n = 14,417 cases). Multivariate Poisson analysis revealed that zip codes with high population density (RR = 2.81), high percentage of farm workers (RR = 1.49), and the composite of those with high and medium percentage of poverty and those with high and medium percentage of non-white residents (RR = 1.59) increased the likelihood of the occurrence of acute urticaria. Conclusion: High population density, farm work, poverty and minority status is associated with a high risk of having acute urticaria.
Prevalence and Predictors of Per- and Polyfluoroalkyl Substances (PFAS) Serum Levels among Members of a Suburban US Volunteer Fire Department
Authors: Graber Judith M. , Black Taylor M. , Shah Nimit N. , Caban-Martinez Alberto J. , Lu Shou-en , Brancard Troy , Yu Chang Ho , Turyk Mary E. , Black Kathleen , Steinberg Michael B. , Fan Zhihua , Burgess Jefferey L.
Published: April 2021

Background: Per-and polyfluoroalkyl substances (PFAS), are ubiquitous pollutants associated with adverse health outcomes. High PFAS levels have been demonstrated among career firefighters; less is known about PFAS levels among volunteer firefighters who comprise two-thirds of US firefighters. Methods: Volunteer fire department members completed a survey and provided blood samples. We calculated geometric means and 95% CIs for PFAS reported by the National Health and Nutrition Examination Survey (NHANES). We compared PFAS distribution and levels among non-Hispanic white adult male study participants to those in the 2015–2016 and 2017–2018 NHANES cycles. We assessed associations between PFAS serum levels and years of firefighting controlling demographics and occupation using linear regression. Results: Participant’s average age was 46.6 years (sd. 17.1). Perfluorododecanoic acid (PFDoA) was detected in almost half study but <3% of NHANES participants; serum levels of PFDoA, perfluorononanoic acid (PFNA), and perfluorodecanoic acid (PFDA) were elevated among participants compared with NHANES. Serum levels of both PFDA and PFDoA were positively associated with years of firefighting. Conclusions: Volunteer firefighters may have a different serum profile and levels of PFAS than the general population. Future work in this area should include volunteer firefighters from other geographic locations and assess sources of PFAS exposure.
Acute Moderate Hypoxia Reduces One-Legged Cycling Performance Despite Compensatory Increase in Peak Cardiac Output: A Pilot Study
Authors: Gatterer Hannes , Menz Verena , Burtscher Martin
Published: April 2021

In severe hypoxia, single-leg peak oxygen uptake (VO2peak) is reduced mainly due to the inability to increase cardiac output (CO). Whether moderate altitude allows CO to increase during single-leg cycling, thereby restoring VO2peak, has not been extensively investigated. Five healthy subjects performed an incremental, maximal, two-legged cycle ergometer test, and on separate days a maximal incremental one-leg cycling test in normoxia and in moderate hypoxia (fraction of inspired oxygen (FiO2) = 15%). Oxygen uptake, heart rate, blood pressure responses, power output, and CO (PhysioFlow) were measured during all tests. Moderate hypoxia lowered single-leg peak power output (154 ± 31 vs. 128 ± 26 watts, p = 0.03) and oxygen uptake (VO2) (36.8 ± 6.6 vs. 33.9 ± 6.9 mL/min/kg, p = 0.04), despite higher peak CO (16.83 ± 3.10 vs. 18.96 ± 3.59 L/min, p = 0.04) and systemic oxygen (O2) delivery (3.37 ± 0.84 vs. 3.47 ± 0.89 L/min, p = 0.04) in hypoxia compared to normoxia. Arterial–venous O2 difference (a–vDO2) was lower in hypoxia (137 ± 21 vs. 112 ± 19 mL/l, p = 0.03). The increases in peak CO from normoxia to hypoxia were negatively correlated with changes in mean arterial pressure (MABP) (p < 0.05). These preliminary data indicate that the rise in CO was not sufficient to prevent single-leg performance loss at moderate altitude and that enhanced baroreceptor activity might limit CO increases in acute hypoxia, likely by reducing sympathetic activation. Since the systemic O2 delivery was enhanced and the calculated a–vDO2 reduced in moderate hypoxia, a potential diffusion limitation cannot be excluded.
Differences between Chinese Adolescent Immigrants and Adolescent Non-Immigrants in Hong Kong: Perceived Psychosocial Attributes, School Environment and Characteristics of Hong Kong Adolescents
Authors: Jiang Hechao , Shek Daniel T. L. , Law Moon Y. M.
Published: April 2021

Although the impact of immigration on adolescent developmental outcomes has received extensive scholarly attention, the impact of internal migration, particularly in the Chinese context, on adolescents’ psychosocial development has not been scientifically investigated. This study examined whether mainland Chinese adolescent immigrants (N = 590) and adolescent non-immigrants (n = 1798) differed on: (a) psychosocial attributes indexed by character traits, well-being, social behavior, and views on child development, (b) perceived school environment, and (c) perceptions of characteristics of Hong Kong adolescents. Consistent with the healthy migration hypothesis, Hong Kong adolescents and mainland Chinese adolescent immigrants did not differ on most of the outcomes; Chinese adolescent immigrants showed higher perceived moral character, empathy, and social trust than did Hong Kong adolescent non-immigrants. Chinese adolescent immigrants also showed more favorable perceptions of the school environment and moral character, social trust and social responsibility of adolescents in Hong Kong. This pioneer Chinese study provides support for the healthy immigration hypothesis (immigration paradox hypothesis) but not the immigration morbidity hypothesis within the specific sociocultural context of Hong Kong in China.
Impact of Individual, Familial and Parental Factors on Adolescent Smoking in Turkey
Authors: Oztekin Coskun , Batra Mehak , Abdelsalam Shady , Sengezer Tijen , Ozkara Adem , Erbas Bircan
Published: April 2021

The burden of adolescent cigarette smoking is substantial. We assess mothers’ and fathers’ attitudes and behaviours on adolescent smoking using a cross-sectional study of n = 707 adolescents. Associations between parental attitudes and behaviours in adolescent smoking were assessed using logistic regression separately for boys and girls. Occasional alcohol use by both parents increased odds of smoking once a day (OR = 2.44, 95% CI 1.26, 4.71, OR = 1.51, 95% CI 0.97, 2.35, respectively). Fathers smoking increased odds for girls (OR = 1.59, 95% CI 1.01, 2.52). A democratic mother decreased odds for boys (OR = 0.32, 95% CI 0.10, 0.93) whereas a protective, demanding mother increased the odds for girls (OR = 8.65, 95% CI 1.38, 54.22). Public health smoking prevention programs could support changing parental behaviours and attitudes in early years to address this burden in countries with authoritarian parenting styles.
A Highly Selective In Vitro JNK3 Inhibitor, FMU200, Restores Mitochondrial Membrane Potential and Reduces Oxidative Stress and Apoptosis in SH-SY5Y Cells
Authors: Rehfeldt Stephanie Cristine Hepp , Laufer Stefan , Goettert Márcia Inês
Published: April 2021

Current treatments for neurodegenerative diseases (ND) are symptomatic and do not affect disease progression. Slowing this progression remains a crucial unmet need for patients and their families. c-Jun N-terminal kinase 3 (JNK3) are related to several ND hallmarks including apoptosis, oxidative stress, excitotoxicity, mitochondrial dysfunction, and neuroinflammation. JNK inhibitors can play an important role in addressing neuroprotection. This research aims to evaluate the neuroprotective, anti-inflammatory, and antioxidant effects of a synthetic compound (FMU200) with known JNK3 inhibitory activity in SH-SY5Y and RAW264.7 cell lines. SH-SY5Y cells were pretreated with FMU200 and cell damage was induced by 6-hydroxydopamine (6-OHDA) or hydrogen peroxide (H2O2). Cell viability and neuroprotective effect were assessed with an MTT assay. Flow cytometric analysis was performed to evaluate cell apoptosis. The H2O2-induced reactive oxygen species (ROS) generation and mitochondrial membrane potential (ΔΨm) were evaluated by DCFDA and JC-1 assays, respectively. The anti-inflammatory effect was determined in LPS-induced RAW264.7 cells by ELISA assay. In undifferentiated SH-SY5Y cells, FMU200 decreased neurotoxicity induced by 6-OHDA in approximately 20%. In RA-differentiated cells, FMU200 diminished cell death in approximately 40% and 90% after 24 and 48 h treatment, respectively. FMU200 reduced both early and late apoptotic cells, decreased ROS levels, restored mitochondrial membrane potential, and downregulated JNK phosphorylation after H2O2 exposure. In LPS-stimulated RAW264.7 cells, FMU200 reduced TNF-α levels after a 3 h treatment. FMU200 protects neuroblastoma SH-SY5Y cells against 6-OHDA- and H2O2-induced apoptosis, which may result from suppressing the JNK pathways. Our findings show that FMU200 can be a useful candidate for the treatment of neurodegenerative disorders.
Analysis of Missense Variants in the Human Histamine Receptor Family Reveals Increased Constitutive Activity of E4106.30×30K Variant in the Histamine H1 Receptor
Authors: Ma Xiaoyuan , Segura Marta Arimont , Zarzycka Barbara , Vischer Henry F. , Leurs Rob
Published: April 2021

The Exome Aggregation Consortium has collected the protein-encoding DNA sequences of almost 61,000 unrelated humans. Analysis of this dataset for G protein-coupled receptor (GPCR) proteins (available at GPCRdb) revealed a total of 463 naturally occurring genetic missense variations in the histamine receptor family. In this research, we have analyzed the distribution of these missense variations in the four histamine receptor subtypes concerning structural segments and sites important for GPCR function. Four missense variants R1273.52×52H, R13934.57×57H, R4096.29×29H, and E4106.30×30K, were selected for the histamine H1 receptor (H1R) that were hypothesized to affect receptor activity by interfering with the interaction pattern of the highly conserved D(E)RY motif, the so-called ionic lock. The E4106.30×30K missense variant displays higher constitutive activity in G protein signaling as compared to wild-type H1R, whereas the opposite was observed for R1273.52×52H, R13934.57×57H, and R4096.29×29H. The E4106.30×30K missense variant displays a higher affinity for the endogenous agonist histamine than wild-type H1R, whereas antagonist affinity was not affected. These data support the hypothesis that the E4106.30×30K mutation shifts the equilibrium towards active conformations. The study of these selected missense variants gives additional insight into the structural basis of H1R activation and, moreover, highlights that missense variants can result in pharmacologically different behavior as compared to wild-type receptors and should consequently be considered in the drug discovery process.
Improved CRISPR/Cas9 Tools for the Rapid Metabolic Engineering of Clostridium acetobutylicum
Authors: Wilding-Steele Tom , Ramette Quentin , Jacottin Paul , Soucaille Philippe
Published: April 2021

Clustered regularly interspaced short palindromic repeats (CRISPR)/Cas (CRISPR-associated proteins)9 tools have revolutionized biology—several highly efficient tools have been constructed that have resulted in the ability to quickly engineer model bacteria, for example, Escherichia coli. However, the use of CRISPR/Cas9 tools has lagged behind in non-model bacteria, hampering engineering efforts. Here, we developed improved CRISPR/Cas9 tools to enable efficient rapid metabolic engineering of the industrially relevant bacterium Clostridium acetobutylicum. Previous efforts to implement a CRISPR/Cas9 system in C. acetobutylicum have been hampered by the lack of tightly controlled inducible systems along with large plasmids resulting in low transformation efficiencies. We successfully integrated the cas9 gene from Streptococcuspyogenes into the genome under control of the xylose inducible system from Clostridium difficile, which we then showed resulted in a tightly controlled system. We then optimized the length of the editing cassette, resulting in a small editing plasmid, which also contained the upp gene in order to rapidly lose the plasmid using the upp/5-fluorouracil counter-selection system. We used this system to perform individual and sequential deletions of ldhA and the ptb-buk operon.
Nicotinamide Phosphoribosyltransferase as a Key Molecule of the Aging/Senescence Process
Authors: Khaidizar Fiqri D. , Bessho Yasumasa , Nakahata Yasukazu
Published: April 2021

Aging is a phenomenon underlined by complex molecular and biochemical changes that occur over time. One of the metabolites that is gaining strong research interest is nicotinamide adenine dinucleotide, NAD+, whose cellular level has been shown to decrease with age in various tissues of model animals and humans. Administration of NAD+ precursors, nicotinamide mononucleotide (NMN) and nicotinamide riboside (NR), to supplement NAD+ production through the NAD+ salvage pathway has been demonstrated to slow down aging processes in mice. Therefore, NAD+ is a critical metabolite now understood to mitigate age-related tissue function decline and prevent age-related diseases in aging animals. In human clinical trials, administration of NAD+ precursors to the elderly is being used to address systemic age-associated physiological decline. Among NAD+ biosynthesis pathways in mammals, the NAD+ salvage pathway is the dominant pathway in most of tissues, and NAMPT is the rate limiting enzyme of this pathway. However, only a few activators of NAMPT, which are supposed to increase NAD+, have been developed so far. In this review, we will focus on the importance of NAD+ and the possible application of an activator of NAMPT to promote successive aging.
Genetic Labeling of Cells Allows Identification and Tracking of Transgenic Platelets in Mice
Authors: Krüger Irena , Reusswig Friedrich , Krott Kim Jürgen , Lersch Celina Fabienne , Spelleken Martina , Elvers Margitta
Published: April 2021

Background: The use of knock-out mouse models is crucial to understand platelet activation and aggregation. Methods: Analysis of the global double fluorescent Cre reporter mouse mT/mG that has been crossbred with the megakaryocyte/platelet specific PF4-Cre mouse. Results: Platelets show bright mT (PF4-Cre negative) and mG (PF4-Cre positive) fluorescence. However, a small proportion of leukocytes was positive for mG fluorescence in PF4-Cre positive mice. In mT/mG;PF4-Cre mice, platelets, and megakaryocytes can be tracked by their specific fluorescence in blood smear, hematopoietic organs and upon thrombus formation. No differences in platelet activation and thrombus formation was observed between mT/mG;PF4-Cre positive and negative mice. Furthermore, hemostasis and in vivo thrombus formation was comparable between genotypes as analyzed by intravital microscopy. Transplantation studies revealed that bone marrow of mT/mG;PF4-Cre mice can be transferred to C57BL/6 mice. Conclusions: The mT/mG Cre reporter mouse is an appropriate model for real-time visualization of platelets, the analysis of cell morphology and the identification of non-recombined platelets. Thus, mT/mG;PF4-Cre mice are important for the analysis of platelet-specific knockout mice. However, a small proportion of leukocytes exhibit mG fluorescence. Therefore, the analysis of platelets beyond hemostasis and thrombosis should be critically evaluated when recombination of immune cells is increased.
Differential Cytotoxicity Mechanisms of Copper Complexed with Disulfiram in Oral Cancer Cells
Authors: Chen Ssu-Yu , Chang Yung-Lung , Liu Shu-Ting , Chen Gunng-Shinng , Lee Shiao-Pieng , Huang Shih-Ming
Published: April 2021

Disulfiram (DSF), an irreversible aldehyde dehydrogenase inhibitor, is being used in anticancer therapy, as its effects in humans are known and less adverse than conventional chemotherapy. We explored the potential mechanism behind the cytotoxicity of DSF-Cu+/Cu2+ complexes in oral epidermoid carcinoma meng-1 (OECM-1) and human gingival epithelial Smulow-Glickman (SG) cells. Exposure to CuCl2 or CuCl slightly but concentration-dependently decreased cell viability, while DSF-Cu+/Cu2+ induced cell death in OECM-1 cells, but not SG cells. DSF-Cu+/Cu2+ also increased the subG1 population and decreased the G1, S, and G2/M populations in OECM-1 cells, but not SG cells, and suppressed cell proliferation in both OECM-1 and SG cells. ALDH enzyme activity was inhibited by CuCl and DSF-Cu+/Cu2+ in SG cells, but not OECM-1 cells. ROS levels and cellular senescence were increased in DSF-Cu+/Cu2+-treated OECM-1 cells, whereas they were suppressed in SG cells. DSF-Cu+/Cu2+ induced mitochondrial fission in OECM-1 cells and reduced mitochondrial membrane potential. CuCl2 increased but DSF- Cu2+ impaired oxygen consumption rates and extracellular acidification rates in OECM-1 cells. CuCl2 stabilized HIF-1α expression under normoxia in OECM-1 cells, and complex with DSF enhanced that effect. Levels of c-Myc protein and its phosphorylation at Tyr58 and Ser62 were increased, while levels of the N-terminal truncated form (Myc-nick) were decreased in DSF-Cu+/Cu2-treated OECM-1 cells. These effects were all suppressed by pretreatment with the ROS scavenger NAC. Overexpression of c-Myc failed to induce HIF-1α expression. These findings provide novel insight into the potential application of DSF-CuCl2 complex as a repurposed agent for OSCC cancer therapy.
In Silico Identification of Small Molecules as New Cdc25 Inhibitors through the Correlation between Chemosensitivity and Protein Expression Pattern
Authors: Lauria Antonino , Martorana Annamaria , La Monica Gabriele , Mannino Salvatore , Mannino Giuseppe , Peri Daniele , Gentile Carla
Published: April 2021

The cell division cycle 25 (Cdc25) protein family plays a crucial role in controlling cell proliferation, making it an excellent target for cancer therapy. In this work, a set of small molecules were identified as Cdc25 modulators by applying a mixed ligand-structure-based approach and taking advantage of the correlation between the chemosensitivity of selected structures and the protein expression pattern of the proposed target. In the first step of the in silico protocol, a set of molecules acting as Cdc25 inhibitors were identified through a new ligand-based protocol and the evaluation of a large database of molecular structures. Subsequently, induced-fit docking (IFD) studies allowed us to further reduce the number of compounds biologically screened. In vitro antiproliferative and enzymatic inhibition assays on the selected compounds led to the identification of new structurally heterogeneous inhibitors of Cdc25 proteins. Among them, J3955, the most active inhibitor, showed concentration-dependent antiproliferative activity against HepG2 cells, with GI50 in the low micromolar range. When J3955 was tested in cell-cycle perturbation experiments, it caused mitotic failure by G2/M-phase cell-cycle arrest. Finally, Western blotting analysis showed an increment of phosphorylated Cdk1 levels in cells exposed to J3955, indicating its specific influence in cellular pathways involving Cdc25 proteins.
Plant Extracellular Vesicles and Nanovesicles: Focus on Secondary Metabolites, Proteins and Lipids with Perspectives on Their Potential and Sources
Authors: Woith Eric , Guerriero Gea , Hausman Jean-Francois , Renaut Jenny , Leclercq Céline C. , Weise Christoph , Legay Sylvain , Weng Alexander , Melzig Matthias F.
Published: April 2021

While human extracellular vesicles (EVs) have attracted a big deal of interest and have been extensively characterized over the last years, plant-derived EVs and nanovesicles have earned less attention and have remained poorly investigated. Although a series of investigations already revealed promising beneficial health effects and drug delivery properties, adequate (pre)clinical studies are rare. This fact might be caused by a lack of sources with appropriate qualities. Our study introduces plant cell suspension culture as a new and well controllable source for plant EVs. Plant cells, cultured in vitro, release EVs into the growth medium which could be harvested for pharmaceutical applications. In this investigation we characterized EVs and nanovesicles from distinct sources. Our findings regarding secondary metabolites indicate that these might not be packaged into EVs in an active manner but enriched in the membrane when lipophilic enough, since apparently lipophilic compounds were associated with nanovesicles while more hydrophilic structures were not consistently found. In addition, protein identification revealed a possible explanation for the mechanism of EV cell wall passage in plants, since cell wall hydrolases like 1,3-β-glucosidases, pectinesterases, polygalacturonases, β-galactosidases and β-xylosidase/α-L-arabinofuranosidase 2-like are present in plant EVs and nanovesicles which might facilitate cell wall transition. Further on, the identified proteins indicate that plant cells secrete EVs using similar mechanisms as animal cells to release exosomes and microvesicles.
Heparan Sulfate Deficiency in Cartilage: Enhanced BMP-Sensitivity, Proteoglycan Production and an Anti-Apoptotic Expression Signature after Loading
Authors: Gerstner Matthias , Severmann Ann-Christine , Chasan Safak , Vortkamp Andrea , Richter Wiltrud
Published: April 2021

Osteoarthritis (OA) represents one major cause of disability worldwide still evading efficient pharmacological or cellular therapies. Severe degeneration of extracellular cartilage matrix precedes the loss of mobility and disabling pain perception in affected joints. Recent studies showed that a reduced heparan sulfate (HS) content protects cartilage from degradation in OA-animal models of joint destabilization but the underlying mechanisms remained unclear. We aimed to clarify whether low HS-content alters the mechano-response of chondrocytes and to uncover pathways relevant for HS-related chondro-protection in response to loading. Tissue-engineered cartilage with HS-deficiency was generated from rib chondrocytes of mice carrying a hypomorphic allele of Exostosin 1 (Ext1), one of the main HS-synthesizing enzymes, and wildtype (WT) littermate controls. Engineered cartilage matured for 2 weeks was exposed to cyclic unconfined compression in a bioreactor. The molecular loading response was determined by transcriptome profiling, bioinformatic data processing, and qPCR. HS-deficient chondrocytes expressed 3–6% of WT Ext1-mRNA levels. Both groups similarly raised Sox9, Col2a1 and Acan levels during maturation. However, HS-deficient chondrocytes synthesized and deposited 50% more GAG/DNA. TGFβ and FGF2-sensitivity of Ext1gt/gt chondrocytes was similar to WT cells but their response to BMP-stimulation was enhanced. Loading induced similar activation of mechano-sensitive ERK and P38-signaling in WT and HS-reduced chondrocytes. Transcriptome analysis reflected regulation of cell migration as major load-induced biological process with similar stimulation of common (Fosl1, Itgα5, Timp1, and Ngf) as well as novel mechano-regulated genes (Inhba and Dhrs9). Remarkably, only Ext1-hypomorphic cartilage responded to loading by an expression signature of negative regulation of apoptosis with pro-apoptotic Bnip3 being selectively down-regulated. HS-deficiency enhanced BMP-sensitivity, GAG-production and fostered an anti-apoptotic expression signature after loading, all of which may protect cartilage from load-induced erosion.
Relationship between NAFLD and Periodontal Disease from the View of Clinical and Basic Research, and Immunological Response
Authors: Hatasa Masahiro , Yoshida Sumiko , Takahashi Hirokazu , Tanaka Kenichi , Kubotsu Yoshihito , Ohsugi Yujin , Katagiri Takaharu , Iwata Takanori , Katagiri Sayaka
Published: April 2021

Periodontal disease is an inflammatory disease caused by pathogenic oral microorganisms that leads to the destruction of alveolar bone and connective tissues around the teeth. Although many studies have shown that periodontal disease is a risk factor for systemic diseases, such as type 2 diabetes and cardiovascular diseases, the relationship between nonalcoholic fatty liver disease (NAFLD) and periodontal disease has not yet been clarified. Thus, the purpose of this review was to reveal the relationship between NAFLD and periodontal disease based on epidemiological studies, basic research, and immunology. Many cross-sectional and prospective epidemiological studies have indicated that periodontal disease is a risk factor for NAFLD. An in vivo animal model revealed that infection with periodontopathic bacteria accelerates the progression of NAFLD accompanied by enhanced steatosis. Moreover, the detection of periodontopathic bacteria in the liver may demonstrate that the bacteria have a direct impact on NAFLD. Furthermore, Porphyromonas gingivalis lipopolysaccharide induces inflammation and accumulation of intracellular lipids in hepatocytes. Th17 may be a key molecule for explaining the relationship between periodontal disease and NAFLD. In this review, we attempted to establish that oral health is essential for systemic health, especially in patients with NAFLD.
Identification of QTLs and a Candidate Gene for Reducing Pre-Harvest Sprouting in Aegilops tauschii–Triticum aestivum Chromosome Segment Substitution Lines
Authors: He Jie , Zhang Dale , Chen Xian , Li Yuge , Hu Minjie , Sun Shaoguang , Su Qing , Su Yarui , Li Suoping
Published: April 2021

Wheat pre-harvest sprouting (PHS) causes serious losses in wheat yield. In this study, precise mapping was carried out in the chromosome segment substitution lines (CSSL) F2 population generated by a direct cross of Zhoumai 18 (PHS-sensitive) and Aegilops tauschii accession T093 (highly PHS-resistant). Three Ae. tauschii-derived quantitative trait loci (QTLs), QDor.3D.1, QDor.3D.2, and QDor.3D.3, were detected on chromosome 3DL using four simple sequence repeats (SSR) markers and 10 developed Kompetitive allele-specific PCR (KASP) markers. Alongside these QTL results, the RNA-Seq and qRT-PCR analysis revealed expression levels of TraesCS3D01G466100 in the QDor.3D.2 region that were significantly higher in CSSLs 495 than in Zhoumai 18 during the seed imbibition treatment. The cDNA sequencing results of TraesCS3D01G466100 showed two single nucleotide polymorphisms (SNPs), resulting in two changed amino acid substitutions between Zhoumai 18 and line 495, and the 148 nt amino acid substitution of TraesCS3D01G466100, derived from Ae. tauschii T093, which may play an important role in the functioning of ubiquitin ligase enzymes 3 (E3) according to the homology protein analysis, which could lead to differential PHS-resistance phenotypes. Taken together, our results may foster a better understanding of the mechanism of PHS resistance and are potentially valuable for marker-assisted selection in practical wheat breeding efforts.
A Consistent and Implicit Rhie–Chow Interpolation for Drag Forces in Coupled Multiphase Solvers
Authors: Hanimann Lucian , Mangani Luca , Darwish Marwan , Casartelli Ernesto , Vogt Damian M.
Published: April 2021

The use of coupled algorithms for single fluid flow simulation has proven its superiority as opposed to segregated algorithms, especially in terms of robustness and performance. In this paper, the coupled approach is extended for the simulation of multi-fluid flows, using a collocated and pressure-based finite volume discretization technique with a Eulerian–Eulerian model. In this context a key ingredient in this method is extending the Rhie–Chow interpolation technique to account for the unique flow coupling that arises from inter-phase drag. The treatment of this inter-fluid coupling and the fashion in which it interacts with the velocity-pressure solution algorithm is presented in detail and its effect on robustness and accuracy is demonstrated using 2D dilute gas–solid flow test case. The results achieved with this technique show substantial improvement in accuracy and performance when compared to a leading commercial code for a transonic nozzle configuration.
Student Thoughts on Virtual Reality in Higher Education—A Survey Questionnaire
Authors: Cicek Igor , Bernik Andrija , Tomicic Igor
Published: April 2021

This paper explores the benefits of using Virtual Reality (VR) technologies in higher education. The theoretical part investigates the classical education system and its features in order to compare advantages of using VR systems in education. VR technologies and its current state in industry and in education were explored in addition to which branches of higher education use these systems. A survey was conducted through an online questionnaire where respondents (N = 55) gave their opinion on VR and the implementation of VR technologies in education. Three hypotheses related to the use of VR technology, student interest, and learning outcomes as well as the effectiveness, immersiveness and the effect of VR systems on the users were tested through 27 questions.
Realising a Push Button Modality for Video-Based Forensics
Authors: Zawali Bako , Ikuesan Richard A. , Kebande Victor R. , Furnell Steven , A-Dhaqm Arafat
Published: April 2021

Complexity and sophistication among multimedia-based tools have made it easy for perpetrators to conduct digital crimes such as counterfeiting, modification, and alteration without being detected. It may not be easy to verify the integrity of video content that, for example, has been manipulated digitally. To address this perennial investigative challenge, this paper proposes the integration of a forensically sound push button forensic modality (PBFM) model for the investigation of the MP4 video file format as a step towards automated video forensic investigation. An open-source multimedia forensic tool was developed based on the proposed PBFM model. A comprehensive evaluation of the efficiency of the tool against file alteration showed that the tool was capable of identifying falsified files, which satisfied the underlying assertion of the PBFM model. Furthermore, the outcome can be used as a complementary process for enhancing the evidence admissibility of MP4 video for forensic investigation.
A Bizarre Planthopper Nymph (Hemiptera: Fulgoroidea) from Mid-Cretaceous Kachin Amber
Authors: Luo Cihang , Wang Bo , Jarzembowski Edmund A.
Published: April 2021

The fossil record of adult planthoppers is comparatively rich, but nymphs are rare and not well studied. Here, we describe a bizarre armoured planthopper nymph, Spinonympha shcherbakovi gen. et sp. nov., in mid-Cretaceous Kachin amber. The new genus is characterized by its large size, body armed with spines and tubercles, extremely long rostrum reaching well beyond the apex of the abdomen; profemur and mesofemur subcylindrical, covered with setae; protibia and mesotibia subquadrangular, densely covered with setae; protarsus and mesotarsus with two segments, tarsomere II longer and wider than I; metatrochanter swollen, metafemur subcylindrical, covered with short setae; metatibia subquadrangular, densely covered with short setae, without lateral spine and pectens without setae; metatarsus with three segments, and metatarsomere III extremely small. The fossil nymph cannot be attributed to any known planthopper family, but can be excluded from many families due to its large size and leg structure. The armoured body was probably developed for defence, and the extremely long rostrum indicates that, in the past, feeding on trees with thick and rough bark was more widespread than today. These features indicate that the new specimen represents a new armoured morphotype of planthopper nymph from the fossil record.
Molecular Characteristics of Fat Body Protein 1 in the Oriental Fruit Fly, Bactrocera dorsalis
Authors: Yu Yao-Chih , Lu Hsuan , Chiang Yi-Cheng , Tsai Cheng-Lung , Zuo Yu-Han , Chen Mei-Er
Published: April 2021

Bactrocera dorsails fat body protein 1 (Bdfbp1) cDNA was cloned (GenBank accession no. MT514270), and the complete 3,749-bp cDNA encoded a 1,152-amino acid protein. The phylogenetic relationship of dipteran fbp1s was analyzed. The sequence XP_028900815 from the insect genome project for Zeugodacus cucurbitae (LOC105219342) was proposed that two fbp1 genes were present in the sequence. The developmental transcriptional expression profiles were determined. In the larval stages, Bdfbp1 mRNA had significantly higher expression in the late third instar larvae compared with first, second, and early third instar larvae. In the pupal stages, the highest expression of Bdfbp1 mRNA was found in the newly pupated pupae and then decreased with age. In the fat body of female adults, Bdfbp1 was highly expressed in newly emerged samples and decreased rapidly over the following three days. In the fat body of male adults, Bdfbp1 was highly expressed in newly eclosed samples. RNAi treatment decreased the expression level of Bdfbp1 without statistical difference. However, RNAi treatment significantly decreased the rate of eclosion. These results suggest that Bdfbp1 may function as a storage protein and be associated with adult eclosion.
Vaginal Probiotics for Reproductive Health and Related Dysbiosis: Systematic Review and Meta-Analysis
Authors: López-Moreno Ana , Aguilera Margarita
Published: April 2021

The use of probiotics in reproductive-related dysbiosis is an area of continuous progress due to the growing interest from clinicians and patients suffering from recurrent reproductive microbiota disorders. An imbalance in the natural colonization sites related to reproductive health—vaginal, cervicovaginal, endometrial, and pregnancy-related altered microbiota—could play a decisive role in reproductive outcomes. Oral and vaginal administrations are in continuous discussion regarding the clinical effects pursued, but the oral route is used and studied more often despite the need for further transference to the colonization site. The aim of the present review was to retrieve the standardized protocols of vaginal probiotics commonly used for investigating their microbiota modulation capacities. Most of the studies selected focused on treating bacterial vaginosis (BV) as the most common dysbiosis; a few studies focused on vulvovaginal candidiasis (VVC) and on pretreatment during in vitro fertilization (IVF). Vaginal probiotic doses administered were similar to oral probiotics protocols, ranging from ≥107 CFU/day to 2.5 × 1010 CFU/day, but were highly variable regarding the treatment duration timing. Moderate vaginal microbiota modulation was achieved; the relative abundance of abnormal microbiota decreased and Lactobacillus species increased.