bluuebunny/crossref_metadata_2025 huggingface dataset:
The Landscape of Outpatient Palliative Care in Germany: Results from a Retrospective Analysis of 14,792 Patients
Authors: Loosen Sven H. , Krieg Sarah , Eschrich Johannes , Luedde Mark , Krieg Andreas , Schallenburger Manuela , Schwartz Jacqueline , Neukirchen Martin , Luedde Tom , Kostev Karel , Roderburg Christoph
Published: November 2022

Background: Palliative care comprises multiprofessional, integrated, person-centered healthcare services for patients and their families facing problems related to progressive or advanced diseases and limited life expectancy. Although non-oncology patients’ needs are similar to those of tumor patients, they are often underestimated. The purpose of our study was to investigate the actual utilization of palliative care services in Germany, especially in the outpatient setting. Methods: Using the IQVIA Disease Analyzer database, a total of 14,792 outpatients from 805 primary care practices in Germany with documented palliative care and related diagnosis between 2018 and 2021 were analyzed. Proportions of different diagnoses among patients receiving outpatient palliative care were stratified by gender and different age groups. Results: The most common underlying diagnosis for outpatient palliative care was cancer (55%), followed by heart failure (16%) and dementia (8%), with age- and sex-specific differences found in the proportion of diagnoses for utilization. While the relative proportions of cancers decreased with age (87% in the 18- to 50-year-old age group versus 37% in the 80-plus age group), the proportion of palliative care related to heart failure increased in the older population (2% in the 18- to 50-year-old age group versus 25% in the 80-plus age group). Conclusions: This study provides an overview of the situation of outpatient palliative care in Germany and shows age- and gender-specific trends regarding the underlying medical diagnoses. Based on these data, palliative care should be adapted to current demographic developments.
COVID-19 in Pregnancy: Knowledge about the Vaccine and the Effect of the Virus. Reliability and Results of the MAMA-19 Questionnaire
Authors: Mannocci Alice , Scaglione Claudia , Casella Giovanna , Lanzone Antonio , La Torre Giuseppe
Published: November 2022

Background: Fear or mistrust of the vaccine and concern for the well-being of their unborn infants are the main reasons for COVID-19 vaccine hesitancy in pregnant women. The aim of this work was to validate a questionnaire on knowledge about COVID-19 vaccination during pregnancy and to examine the sources of information in a group of new mothers, as well as their effectiveness and intelligibility. Methods: A literature review was carried out to develop a questionnaire of forty-five questions, divided into six sections, called MAMA-19. The assessment of agreement and the interrater reliability was carried out using Cronbach’s analysis and Cohen’s kappa statistic. Data obtained from the questionnaire were analysed using descriptive and univariate statistics. Results: The total alpha values in the two sections about knowledge of vaccination during pregnancy and about the effects of disease and possible post-COVID-19 consequences for the unvaccinated showed sufficient consistency, at 0.860 and 0.725, respectively. Non-vaccinated women thought that receiving the COVID-19 vaccine during pregnancy can lead to malformations in the newborn (60% vs. 40%, p = 0.002) and to an increased risk of foetal growth restriction (61.9% vs. 38.1%, p < 0.001). The percentage of vaccinated women was significantly higher than non-vaccinated when more than one professional was consulted and consistent information was received from them (74.2% vs. 25.8%, p = 0.008). Conclusion: The MAMA-19 questionnaire shows results in line with the literature and valid in the two main sections. It is quick to use for measuring communication effectiveness by healthcare professionals and institutions in the context of the COVID-19 vaccination campaign in the pregnant population. The results evidence that a physician’s recommendation to get vaccinated is the most important factor in maternal decision making, regardless of geographic, social or educational context.
The Factors Influencing Chinese University Teachers’ Intentions for Using the Micro-Lecture in the Post COVID-19 Era
Authors: Xie Qihui
Published: November 2022

The COVID-19 pandemic has produced a far-reaching influence on higher education and the teaching activities of teachers in Chinese universities. The intentions of teachers in universities for using the micro-lecture, one of the educational informationization products, and the influencing factors of the intentions for using micro-lectures, are changing in the post COVID-19 era. This paper, based on the Technical Acceptance Model (TAM) and the Unified Theory of Acceptance and Use of Technology (UTAUT), constructed the research hypotheses for the influence factors of micro-lecture usage intentions of teachers in universities in the post COVID-19 era, and made corresponding verifications through the Structural Equation Model (SEM). As shown by the results therefrom: (1) the micro-lecture usage experience before the outbreak of the COVID-19 pandemic significantly affected the usage intentions of teachers in universities; (2) the perceived usefulness influenced the usage intention directly, but the perceived ease of use did not directly produce influence; (3) policy impact had no significant influence on the perceived usefulness and the perceived ease of use of university teachers for micro-lecture use; (4) social relations and personal innovativeness have significant impacts on perceived usefulness, teaching objectives and micro-lecture characteristics have significant impacts on the perceived ease of use. In this paper, suggestions and opinions on popularizing micro-lecture usage in the post COVID-19 era were put forward on the basis of research conclusions therein.
Effects of Kinesio Taping and Rigid Taping on Gluteus Medius Muscle Activation in Healthy Individuals: A Randomized Controlled Study
Authors: Zaworski Kamil , Baj-Korpak Joanna , Kręgiel-Rosiak Anna , Gawlik Krystyna
Published: November 2022

Background: Gluteus medius muscle (GMed) dysfunctions may be associated with pain and functional problems in the lumbar spine and lower limbs. The study sought to assess the effects of applying kinesio taping (KT) and rigid taping (RT) on GMed in the dominant leg of healthy individuals. Furthermore, an attempt was made to indicate which of the applied exercises brought about the greatest activation of GMed. Methods: The study included 90 individuals, with an average age of 21.79. The participants were randomly assigned to one of three groups: kinesio tape (KT), rigid tape (RT) and placebo tape (C). GMed activation was assessed using sEMG during the performance of such exercises as glute bridge, unilateral glute bridge, clamshell, pelvic drop and lunge. Each of the participants was examined three times—before taping, immediately after and 48 h after taping. Results: Before taping, the greatest GMed activation on the dominant side was noted in clamshell (54.12 %MVIC), whereas the lowest GMed activation was observed in glute bridge (36.35 %MVIC). The comparison of results obtained before and immediately after taping in all the groups revealed a statistically significant increase in GMed activation (p < 0.05), while the comparison of results achieved before and 48 h after taping showed significant differences in glute bridge in groups KT and RT. In all the groups, the differences in results obtained in the other exercises were not significant. Taking into account each of the applied exercises, at none of the three stages of examination were the differences between the groups significant. Conclusions: Regardless of the type of taping applied (KT, RT, C), a significant increase in GMed activation was noted in all the exercises immediately after taping. At none of the stages of examination were the differences between the groups significant.
Relationship between Helicopter Parenting and Chinese Elementary School Child Procrastination: A Mediated Moderation Model
Authors: Zhang Ronghua , Zhang Huanrong , Guo Xiaofeng , Wang Jiali , Zhao Zhongxiang , Feng Lean
Published: November 2022

Background: The family environment is essential for elementary school children’s development. With smartphone penetration into all aspects of people’s lives, how parenting affects children’s behavior may show new patterns. Objective: This study constructed a mediated moderation model, focusing on the mediating role of child self-control and parental phubbing to clarify the relationship between helicopter parenting (over-parenting) and child procrastination and its mechanisms. Methods: The Smartphone Addiction Scale for Chinese Adults, Brief Self-Control Scale, Over-Parenting Questionnaire, and Short General Procrastination Scale were employed to investigate 562 elementary school-age children and their parents. Results: After data analysis, this study showed the following: (1) helicopter parenting was significantly and positively related to child self-control, child procrastination, and parental smartphone use; (2) child self-control partially mediated the relationship between helicopter parenting and child procrastination; and (3) pathways between helicopter parenting and child self-control were moderated by mother-phubbing behavior. Conclusion: These findings inform parents of their roles in family education.
High-Frequency Intraoral Ultrasound for Preoperative Assessment of Depth of Invasion for Early Tongue Squamous Cell Carcinoma: Radiological–Pathological Correlations
Authors: Caprioli Simone , Casaleggio Alessandro , Tagliafico Alberto Stefano , Conforti Cristina , Borda Fabio , Fiannacca Martina , Filauro Marta , Iandelli Andrea , Marchi Filippo , Parrinello Giampiero , Peretti Giorgio , Cittadini Giuseppe
Published: November 2022

The eighth edition of the TNM classification officially introduced “depth of invasion” (DOI) as a criterion for determining the T stage in tongue squamous cell carcinoma. The DOI is a well-known independent risk factor for nodal metastases. In fact, several experts strongly suggest elective neck dissection for tongue cancer with a DOI > 4 mm due to the high risk of early and occult nodal metastases. Imaging plays a pivotal role in preoperative assessments of the DOI and, hence, in planning the surgical approach. Intraoral ultrasound (IOUS) has been proposed for early-stage SCC of the oral tongue as an alternative to magnetic resonance imaging (MRI) for local staging. The aim of this work is to investigate the accuracy of IOUS in the assessment of the DOI in early oral SCC (CIS, pT1, and pT2). A total of 41 patients with tongue SCCs (CIS-T2) underwent a preoperative high-frequency IOUS. An IOUS was performed using a small-size, high-frequency hockey-stick linear probe. The ultrasonographic DOI (usDOI) was retrospectively compared to the pathological DOI (pDOI) as the standard reference. In patients who underwent a preoperative MRI, their usDOI, magnetic resonance DOI (mriDOI), and pDOI were compared. Specificity and sensitivity for the IOUS to predict a pDOI > 4 mm and to differentiate invasive and noninvasive tumors were also evaluated. A high correlation was found between the pDOI and usDOI, pDOI and mriDOI, and usDOI and mriDOI (Spearman’s ρ = 0.84, p < 0.0001, Spearman’s ρ = 0.79, p < 0.0001, and Spearman’s ρ = 0.91, p < 0.0001, respectively). A Bland–Altman plot showed a high agreement between the usDOI and pDOI, even though a mean systematic error was found between the usDOI and pDOI (0.7 mm), mriDOI and pDOI (1.6 mm), and usDOI and mriDOI (−0.7 mm). The IOUS was accurate at determining the T stage (p < 0.0001). The sensitivity and specificity for the IOUS to predict a pDOI ≥4 mm were 92.31% and 82.14%, respectively, with an AUC of 0.87 (p < 0.0001). The specificity, sensitivity, negative predictive value (NPV), and positive predictive value (PPV) for the IOUS to predict an invasive cancer were 100%, 94.7%, 60%, and 100%, respectively. The AUC was 0.8 (95% CI 0.646–0.908, p < 0.0001). The IOUS was accurate in a preoperative assessment of a pDOI and T stage, and can be proposed as an alternative to MRI in the preoperative staging of tongue SCC.
Does Time Heal Trauma? 18 Month Follow-Up Study of Syrian Refugees’ Mental Health in Iraq’s Kurdistan Region
Authors: Mahmood Harem Nareeman , Ibrahim Hawkar , Ismail Azad Ali , Neuner Frank
Published: November 2022

The findings of longitudinal studies on traumatized refugees have shown that factors related to premigration, migration, and post-migration experiences determine changes in mental health over time. The primary aim of this follow-up study was to examine the potential change in the prevalence rates of probable PTSD and depression among Syrian refugees in Iraq. An unselected group of N = 92 Syrian adult refugees was recruited from Arbat camps in Sulaymaniyah Governorate in Iraq’s Kurdistan Region, and then interviewed at two different time points between July 2017 and January 2019. Locally validated instruments were used to assess traumatic events and mental health symptoms. The primary results showed no significant change in the mean scores of PTSD and depression symptoms from the first measurement to the second measurement over the course of 18 months. On the individual level, no reliable change was found for either PTSD or depression symptoms in more than three-quarters of the participants (78.3% and 77.2%, respectively). New adversities and traumatic events that occurred over the 18 months between the interviews were a significant predictor of increasing trauma-related symptoms. After the flight from conflict settings, trauma-related disorders seem to be chronic for the majority of Syrian refugees. Further longitudinal studies are needed in order to identify specific risk factors that lead to maintaining or worsening mental health symptoms over time, and to explore effective therapeutic intervention methods for this traumatized population.
Do Green Finance Policies Foster Environmental, Social, and Governance Performance of Corporate?
Authors: Wang Xingshuai , Elahi Ehsan , Khalid Zainab
Published: November 2022

The green finance policy is crucial for enterprises to participate in environmental governance actively. Taking the “Green Credit Guidelines” issued by China in 2012 as a quasi-natural experiment, this study investigated the impact of green finance policies on corporate environmental, social, and governance (ESG) performance by using a continuous Difference-in-Differences (DID) model based on the data of listed companies from 2006 to 2020. The conclusions are: (1) The green finance policy significantly improves corporate ESG, but the effects vary across enterprises. (2) The policy has encouraged enterprises to develop and adopt green products and technologies. Still, it has not had a positive effect on the treatment of enterprise pollutant emissions because the implementation of the policy makes enterprises pay more attention to front-end risk control than pollution treatment after production. (3) Research results have heterogeneity. The impact of green finance policies on enterprises at different levels of environmental regulation is different. Enterprises in areas with high intensity of environmental regulation are more vulnerable to green credit. The conclusion of this paper helps improve the green finance policy system, enhance the awareness and level of corporate ESG, and strengthen the collaborative governance of policies and enterprises on environmental issues in combination with the mandatory environmental regulations and incentive mechanisms to promote the green development of enterprises and realize the goal of carbon neutrality.
Effects of Yoga Intervention on Functional Movement Patterns and Mindfulness in Collegiate Athletes: A Quasi-Experimental Study
Authors: Xu Dan , Wu Hua , Ruan Hui , Yuan Cunzhu , Gao Junke , Guo Meng
Published: November 2022

High-quality movement patterns and high levels of mindfulness are thought to be beneficial in preventing sports injuries. Yoga is recommended in the field of athlete rehabilitation. This study investigated the effects of yoga intervention on functional movement patterns and mindfulness in collegiate athletes. It is a quasi-experimental study with a pre/post-test control design. The participants were divided into a yoga group and a control group. A Functional Movement Screen and the Mindful Attention Awareness Scale were used to assess participants’ basic movement patterns and mindfulness before and after 12 weeks of yoga intervention (two classes per week, 90 min per class). The results show that the yoga group’s FMS scores improved more compared to the control group [F(1,78) = 29.08, p < 0.001, ŋp2 = 0.27], and that the scores for the deep squat (ŋp2 = 0.4), shoulder mobility (ŋp2 = 0.17), and trunk stability pushup (ŋp2 = 0.36) improved substantially. The dysfunctional score ratio for deep squats (χ2 = 18.57, p < 0.001), shoulder mobility (χ2 = 26.90, p < 0.001), trunk stability pushup (χ2 = 17.07, p < 0.001), and rotatory stability (χ2= 38.29, p <0.001) decreased significantly compared with the control group, but there was no significant improvement in asymmetric movement patterns (χ2 = 0.75, p = 0.39). The mindfulness scores in the yoga group significantly exceeded those of the control group [F(1,78) = 13.56, p < 0.001, ŋp2 = 0.15]. These results suggest that yoga intervention can improve functional movement patterns and mindfulness levels, but further evidence is needed to determine whether yoga could positively influence sports injuries.
Promote Positive Behaviors in Preschoolers by Implementing an Innovative Educational Program for the Training and Development of Social and Emotional Skills (DeCo–S.E.)
Authors: Badau Adela , Trifan Irina-Mihaela
Published: November 2022

The purpose of our research is to determine emotional and behavioural modelling in an emotionally safe environment in a group of kindergarten preschoolers, following the application of an innovative curriculum project, designed and implemented for the first time, Educational Program for the training and development of social and emotional skills (DeCo-S.E.). Eighteen teachers, involved in the research, were divided into two samples: the experimental group (EG) consisting of 10 preschool teachers who were trained to take up the DeCo-S.E. program, and the control group (GC) including eight preschool teachers who applied the classic educational strategies. The DeCo–S.E. program is aimed at developing social and emotional skills (emotion identification, frustration tolerance), reducing behavior problems, and solving problems with peers as part of the training process. The study also included 142 children in their last year at kindergarten, aged X ± SD 5.87 ± 2.87 years old, divided into two groups: the EG consisting of 74 children to whom the experimental program was implemented and the GC comprising 72 children who did not take part in the training program. In the present study, we applied only the Preschool and Kindergarten Behavior Scales for Teachers (PKBS-2) questionnaire to children in the pre- and post-experimental phases. The results were processed with the statistical software SPSS 22. The analysis of the scores of the questionnaire applied to the children highlighted a significant improvement in EG on both scales. The study reveals the effectiveness of the Development of social and emotional skills programs in preschool children has proven its effectiveness by reducing undesirable/maladaptive behaviours and positively developing socio-emotional skills in preschool children.
Chronic Community Exposure to Environmental Metal Mixtures Is Associated with Selected Cytokines in the Navajo Birth Cohort Study (NBCS)
Authors: Thompson González Nicole , Ong Jennifer , Luo Li , MacKenzie Debra
Published: November 2022

Many tribal populations are characterized by health disparities, including higher rates of infection, metabolic syndrome, and cancer—all of which are mediated by the immune system. Members of the Navajo Nation have suffered chronic low-level exposure to metal mixtures from uranium mine wastes for decades. We suspect that such metal and metalloid exposures lead to adverse health effects via their modulation of immune system function. We examined the relationships between nine key metal and metalloid exposures (in blood and urine) with 11 circulating biomarkers (cytokines and CRP in serum) in 231 pregnant Navajo women participating in the Navajo Birth Cohort Study. Biomonitored levels of uranium and arsenic species were considerably higher in participants than NHANES averages. Each biomarker was associated with a unique set of exposures, and arsenic species were generally immunosuppressive (decreased cellular and humoral stimulating cytokines). Overall, our results suggest that environmental metal and metalloid exposures modulate immune status in pregnant Navajo women, which may impact long-term health outcomes in mothers and their children.
Analysis of Landscape Characteristics and Influencing Factors of Residential Areas on the Qinghai–Tibet Plateau: A Case Study of Tibet, China
Authors: Niu Dingwei , Wang Lucang , Qiao Fuwei , Li Wei
Published: November 2022

The Qinghai–Tibet Plateau is the largest ecological barrier and one of the most vulnerable areas of the ecological environmental system. However, the increasing frequency of human activities in the Qinghai–Tibet Plateau has led to strong interference. Residential areas are the main places in which human activities are carried out and, as such, can effectively reflect the intensity of activities. Based on this, this research takes the Tibet Autonomous Region as the study area and analyzes the distribution characteristics of Tibetan residential areas using Zipf’s law and various landscape indices, as well as discussing the influences of altitude, hydrology, ecological environment, and location on residential area distribution. The obtained results indicate the following: (1) The residential areas in Tibet basically conform to the rank–size principle. The residential areas in central and northwest Tibet are concentrated in size distribution, and the relatively large residential areas are prominent, while the residential areas in the eastern Hengduan mountain region are relatively balanced in size distribution. (2) The landscape index results demonstrate that the counties with an unbalanced distribution of residential areas are mainly concentrated in the northwest of Tibet, while the residential areas in the counties and regions where the administrative stations of each prefecture-level city (or region) are located tend to present a polarization phenomenon, with large patches. The area distribution of residential areas showed a “medium–high–low” pattern from southeast to northwest. The residential areas in eastern Tibet have a high degree of fragmentation and a low degree of aggregation, while the residential areas in northwest Tibet have a low degree of fragmentation and a relatively high degree of aggregation. (3) The residential areas in Tibet are most concentrated in the altitude range of 3000–5000 m above sea level and their water affinity and road–affinity are strong, with the distribution of residential areas within 500 m of roads and water networks accounting for more than one-quarter. The vegetation coverage in the residential areas is low, inconsistent with the surface vegetation coverage rate over the whole of Tibet.
Positive Childhood Experiences Associate with Adult Flourishing Amidst Adversity: A Cross Sectional Survey Study with a National Sample of Young Adults
Authors: Yu Zhiyuan , Wang Lin , Chen Wenyi , Zhang Juan , Bettencourt Amie F.
Published: November 2022

The purpose of this study was to examine the prevalence of PCEs among young adults in Mainland China and the extent to which the cumulative number of PCEs moderates the associations between ACEs and flourishing in adulthood. Between August and November 2020, we used convenience and snowball sampling to recruit 9468 young adults, ages 18–35, enrolled in undergraduate or graduate programs at universities in Mainland China to participate in a survey, which included measures on flourishing, exposure to ACEs and PCEs, and demographic characteristics. Approximately 92% of participants reported experiencing seven to nine PCEs, with harmonious family relationships (96.9%), feeling supported by friends (96.8%) and being treated fairly at school (96.3%) being the most common PCEs reported. Results of the multiple regression indicated that the cumulative number of PCEs statistically significantly moderated the relation between the cumulative number of ACEs and flourishing (interaction term b = −0.060 [−0.071, −0.049], p < 0.001, adjusted R2 = 0.183); as the number of ACEs increased up through eight ACEs, decreases in flourishing were smaller among those with higher numbers of PCEs. PCEs are common among young adults from Mainland China and serve a potential buffering effect against exposure to ACEs.
The Phosphorus Transport in Groundwater from Phosphogypsum-Based Cemented Paste Backfill in a Phosphate Mine: A Numerical Study
Authors: Chen Qiusong , Zhou Huibo , Liu Yikai , Wang Daolin
Published: November 2022

Stacked phosphogypsum (PG) can not only cause a waste of resources but also has a serious negative impact on the surface environment. Phosphogypsum backfilling (PGB) in the underground goaf is a useful approach to effectively address the PG environmental problems. However, the effects of this approach on the groundwater environment have not been studied. Therefore, the present study aims to assess the spatiotemporal evolution mechanism of total phosphorus (TP) in groundwater to solve the diffusion regular pattern of TP in PGB bodies, as well as to manage and mitigate the impacts of TP on the groundwater system. In this study, leaching toxicity experiments and a numerical groundwater simulation software (GMS10.4) were combined to develop a three-dimensional conceptual model for predicting the groundwater flow and contaminant transport under steady-state conditions in a phosphorus mine in Anhui. The results showed a lower TP concentration than the TP standard concentration (0.2 mg/L) at a source concentration of 0.59 mg/L. However, groundwater TP source concentrations of 1.88 and 2.46 mg/L in the study area were found to exceed the standard concentration for a certain time and areas. In addition, the transport and dispersion of TP are influenced not only by the groundwater flow field, drainage ditches, rivers, and wells but also by the adsorption and attenuation effects of the soil that occur during the transport process, affecting the dispersion distance and distribution of groundwater TP concentrations. The results of the present study can promote the development of groundwater-friendly PGB technology, providing a great significance to the construction of green mines and the promotion of ecological civilization.
Structure- and Content-Dependent Efficiency of Cas9-Assisted DNA Cleavage in Genome-Editing Systems
Authors: Baranova Svetlana V. , Zhdanova Polina V. , Lomzov Alexander A. , Koval Vladimir V. , Chernonosov Alexander A.
Published: November 2022

Genome-editing systems, being some of the key tools of molecular biologists, represent a reasonable hope for progress in the field of personalized medicine. A major problem with such systems is their nonideal accuracy and insufficient selectivity. The selectivity of CRISPR-Cas9 systems can be improved in several ways. One efficient way is the proper selection of the consensus sequence of the DNA to be cleaved. In the present work, we attempted to evaluate the effect of formed non-Watson–Crick pairs in a DNA duplex on the efficiency of DNA cleavage in terms of the influence of the structure of the formed partially complementary pairs. We also studied the effect of the location of such pairs in DNA relative to the PAM (protospacer-adjacent motif) on the cleavage efficiency. We believe that the stabilization of the Cas9-sgRNA complex with a DNA substrate containing noncomplementary pairs is due to loop reorganization in the RuvC domain of the enzyme. In addition, PAM-proximal mismatches in the DNA substrate lower enzyme efficiency because the “seed” region is involved in binding and cleavage, whereas PAM-distal mismatches have no significant impact on target DNA cleavage. Our data suggest that in the case of short duplexes with mismatches, the stages of recognition and binding of dsDNA substrates by the enzyme determine the reaction rate and time rather than the thermodynamic parameters affected by the “unwinding” of DNA. The results will provide a theoretical basis for predicting the efficiency and accuracy of CRISPR-Cas9 systems at cleaving target DNA.
In-Plant Persistence and Systemic Transport of Nicotiana benthamiana Retrozyme RNA
Authors: Lezzhov Alexander A. , Tolstyko Eugene A. , Atabekova Anastasia K. , Chergintsev Denis A. , Morozov Sergey Y. , Solovyev Andrey G.
Published: November 2022

Retrozymes are nonautonomous retrotransposons with hammerhead ribozymes in their long terminal repeats (LTRs). Retrozyme transcripts can be self-cleaved by the LTR ribozyme, circularized, and can undergo RNA-to-RNA replication. Here, we demonstrate that the Nicotiana benthamiana genome contains hundreds of retrozyme loci, of which nine represent full-length retrozymes. The LTR contains a promoter directing retrozyme transcription. Although retrozyme RNA is easily detected in plants, the LTR region is heavily methylated, pointing to its transcriptional silencing, which can be mediated by 24 nucleotide-long retrozyme-specific RNAs identified in N. benthamiana. A transcriptome analysis revealed that half of the retrozyme-specific RNAs in plant leaves have no exact matches to genomic retrozyme loci, containing up to 13% mismatches with the closest genomic sequences, and could arise as a result of many rounds of RNA-to-RNA replication leading to error accumulation. Using a cloned retrozyme copy, we show that retrozyme RNA is capable of replication and systemic transport in plants. The presented data suggest that retrozyme loci in the N. benthamiana genome are transcriptionally inactive, and that circular retrozyme RNA can persist in cells due to its RNA-to-RNA replication and be transported systemically, emphasizing functional and, possibly, evolutionary links of retrozymes to viroids—noncoding circular RNAs that infect plants.
Effect of the pH on the Antibacterial Potential and Cytotoxicity of Different Plasma-Activated Liquids
Authors: Sampaio Aline da Graça , Chiappim William , Milhan Noala Vicensoto Moreira , Botan Neto Benedito , Pessoa Rodrigo , Koga-Ito Cristiane Yumi
Published: November 2022

In this study, different plasma-activated liquids were evaluated for their antimicrobial effects against Escherichia coli, as well as for their cytotoxicity on mammalian cells. The PALs were prepared from distilled (DIS), deionized (DI), filtered (FIL), and tap (TAP) water. Additionally, 0.9% NaCl saline solution (SAL) was plasma-activated. These PALs were prepared using 5 L/min air gliding arc plasma jet for up to 60.0 min of exposure. Subsequently, the physicochemical properties, such as, the oxidation-reduction potential (ORP), the pH, the conductivity, and the total dissolved solids (TDS) were characterized by a water multiparameter. The PALs obtained showed a drastic decrease in the pH with increasing plasma exposure time, in contrast, the conductivity and TDS increased. In a general trend, the UV-vis analyses identified a higher production of the following reactive species of nitrogen and oxygen (RONS), HNO2, H2O2, NO3−, and NO2−. Except for the plasma-activated filtered water (PAW-FIL), where there was a change in the position of NO2− and NO3− at some pHs, The higher production of HNO2 and H2O2-reactive species was observed at a low pH. Finally, the standardized suspensions of Escherichia coli were exposed to PAL for up to 60.0 min. The plasma-activated deionized water (PAW-DI pH 2.5), plasma-activated distilled water (PAW-DIS pH 2.5 and 3), and plasma-activated tap water (PAW-TAP 3.5) showed the best antimicrobial effects at exposure times of 3.0, 10.0, and 30.0 min, respectively. The MTT analysis demonstrated low toxicity of all of the PAL samples. Our results indicate that the plasma activation of different liquids using the gliding arc system can generate specific physicochemical conditions that produce excellent antibacterial effects for E. coli with a safe application, thus bringing future contributions to creating new antimicrobial protocols.
Conservation of Importin α Function in Apicomplexans: Ivermectin and GW5074 Target Plasmodium falciparum Importin α and Inhibit Parasite Growth in Culture
Authors: Walunj Sujata B. , Wang Chunxiao , Wagstaff Kylie M. , Patankar Swati , Jans David A.
Published: November 2022

Signal-dependent transport into and out of the nucleus mediated by members of the importin (IMP) superfamily of nuclear transporters is critical to the eukaryotic function and a point of therapeutic intervention with the potential to limit disease progression and pathogenic outcomes. Although the apicomplexan parasites Plasmodium falciparum and Toxoplasma gondii both retain unique IMPα genes that are essential, a detailed analysis of their properties has not been performed. As a first step to validate apicomplexan IMPα as a target, we set out to compare the properties of P. falciparum and T. gondii IMPα (PfIMPα and TgIMPα, respectively) to those of mammalian IMPα, as exemplified by Mus musculus IMPα (MmIMPα). Close similarities were evident, with all three showing high-affinity binding to modular nuclear localisation signals (NLSs) from apicomplexans as well as Simian virus SV40 large tumour antigen (T-ag). PfIMPα and TgIMPα were also capable of binding to mammalian IMPβ1 (MmIMPβ1) with high affinity; strikingly, NLS binding by PfIMPα and TgIMPα could be inhibited by the mammalian IMPα targeting small molecules ivermectin and GW5074 through direct binding to PfIMPα and TgIMPα to perturb the α-helical structure. Importantly, GW5074 could be shown for the first time to resemble ivermectin in being able to limit growth of P. falciparum. The results confirm apicomplexan IMPα as a viable target for the development of therapeutics, with agents targeting it worthy of further consideration as an antimalarial.
Molecular Structure, Vibrational Spectrum and Conformational Properties of 4-(4-Tritylphenoxy)phthalonitrile-Precursor for Synthesis of Phthalocyanines with Bulky Substituent
Authors: Tverdova Natalia V. , Giricheva Nina I. , Maizlish Vladimir E. , Galanin Nikolay E. , Girichev Georgiy V.
Published: November 2022

By DFT method with B3LYP, PBE, CAM-B3LYP, and B97D functionals, it was found that the molecule 4-(4-tritylphenoxy)phthalonitrile (TPPN) has four conformers. The geometric structure, vibrational frequencies, electronic characteristics, and thermodynamic functions of conformers, as well as the structure and energy of transition states, were determined. IR spectrum of TPPN film contains vibrational bands belonging to different conformers. The assignment of bands was performed basing the distribution of normal vibration energy on internal coordinates. A synchronous electron diffraction/mass spectrometric experiment was performed to determine the structure of conformers in a saturated TPPN vapor. The elemental composition of the ions recorded in the mass spectrum indicates the thermal stability of TPPN at least up to T = 200 °C. The difference in the structure of tetrasubstituted metal phthalocyanines, which can be synthesized from different TPPN conformers, has been shown.
Exhausted but Not Senescent T Lymphocytes Predominate in Lupus Nephritis Patients
Authors: Lioulios Georgios , Mitsoglou Zoi , Fylaktou Asimina , Xochelli Aliki , Christodoulou Michalis , Stai Stamatia , Moysidou Eleni , Konstantouli Afroditi , Nikolaidou Vasiliki , Papagianni Aikaterini , Stangou Maria
Published: November 2022

Lupus nephritis (LN), a chronic inflammatory disease, is characterized by the substantial disruption of immune homeostasis. This study examines its effects on the T lymphocyte phenotype and, particularly, its senescence- and exhaustion-related immune alterations. T cell subpopulations were determined with flow cytometry in 30 LN patients and 20 healthy controls (HCs), according to the expression of senescence- (CD45RA, CCR7, CD31, CD28, CD57), and exhaustion- (PD1) related markers. The immune phenotype was associated with disease activity and renal histology. LN patients were characterized by pronounced lymphopenia, mainly affecting the CD4 compartment, with a concurrent reduction in the naïve, central and effector memory subsets compared to the HCs. In the CD8 compartment, the naïve subsets were significantly lower than that of the HCs, but a shift in the T cells occurred towards the central memory population. CD4+PD1+ and CD8+PD1+ cells were increased in the LN patients compared to the HCs. However, in CD4 T cells, the increase was limited to CD45RA+, whereas in CD8 T cells, both CD45RA+ and CD45RA− subsets were affected. Disease activity was correlated with CD4+PD1+ and highly differentiated CD4+CD28-CD57+ cells. Histology was only associated with CD4 T cell disturbances, with stage IV presenting reduced naïve and increased senescent subsets. Exhausted T lymphocyte subpopulations predominate within LN patients, while the T cell phenotype varies depending on disease activity.
Allosteric Inhibition of Neutral Sphingomyelinase 2 (nSMase2) by DPTIP: From Antiflaviviral Activity to Deciphering Its Binding Site through In Silico Studies and Experimental Validation
Authors: Álvarez-Fernández Hadrián , Mingo-Casas Patricia , Blázquez Ana-Belén , Caridi Flavia , Saiz Juan Carlos , Pérez-Pérez María-Jesús , Martín-Acebes Miguel A. , Priego Eva-María
Published: November 2022

Flavivirus comprises globally emerging and re-emerging pathogens such as Zika virus (ZIKV), Dengue virus (DENV), and West Nile virus (WNV), among others. Although some vaccines are available, there is an unmet medical need as no effective antiviral treatment has been approved for flaviviral infections. The development of host-directed antivirals (HDAs) targeting host factors that are essential for viral replication cycle offers the opportunity for the development of broad-spectrum antivirals. In the case of flaviviruses, recent studies have revealed that neutral sphingomyelinase 2, (nSMase2), involved in lipid metabolism, plays a key role in WNV and ZIKV infection. As a proof of concept, we have determined the antiviral activity of the non-competitive nSMase2 inhibitor DPTIP against WNV and ZIKV virus. DPTIP showed potent antiviral activity with EC50 values of 0.26 µM and 1.56 µM for WNV and ZIKV, respectively. In order to unravel the allosteric binding site of DPTIP in nSMase2 and the details of the interaction, computational studies have been carried out. These studies have revealed that DPTIP could block the DK switch in nSMase2. Moreover, the analysis of the residues contributing to the binding identified His463 as a crucial residue. Interestingly, the inhibitory activity of DPTIP on the H463A mutant protein supported our hypothesis. Thus, an allosteric cavity in nSMase2 has been identified that can be exploited for the development of new inhibitors with anti-flaviviral activity.
Carbon-Coated ZnS-FeS2 Heterostructure as an Anode Material for Lithium-Ion Battery Applications
Authors: Naveenkumar Perumal , Maniyazagan Munisamy , Kang Nayoung , Yang Hyeon-Woo , Kang Woo-Seung , Kim Sun-Jae
Published: November 2022

The construction of carbon-coated heterostructures of bimetallic sulfide is an effective technique to improve the electrochemical activity of anode materials in lithium-ion batteries. In this work, the carbon-coated heterostructured ZnS-FeS2 is prepared by a two-step hydrothermal method. The crystallinity and nature of carbon-coating are confirmed by the investigation of XRD and Raman spectroscopy techniques. The nanoparticle morphology of ZnS and plate-like morphology of FeS2 is established by TEM images. The chemical composition of heterostructure ZnS-FeS2@C is discovered by an XPS study. The CV results have disclosed the charge storage mechanism, which depends on the capacitive and diffusion process. The BET surface area (37.95 m2g−1) and lower Rct value (137 Ω) of ZnS-FeS2@C are beneficial to attain higher lithium-ion storage performance. It delivered a discharge capacity of 821 mAh g−1 in the 500th continuous cycle @ A g−1, with a coulombic efficiency of around 100%, which is higher than the ZnS-FeS2 heterostructure (512 mAh g−1). The proposed strategy can improve the electrochemical performance and stability of lithium-ion batteries, and can be helpful in finding highly effective anode materials for energy storage devices.
Genome-Wide Identification of the Highly Conserved INDETERMINATE DOMAIN (IDD) Zinc Finger Gene Family in Moso Bamboo (Phyllostachys edulis)
Authors: Guo Xiaoqin , Zhou Minshu , Chen Jiaoyu , Shao Mingxia , Zou Longhai , Ying Yeqing , Liu Shenkui
Published: November 2022

INDETERMINATE DOMAIN (IDD) proteins, a family of transcription factors unique to plants, function in multiple developmental processes. Although the IDD gene family has been identified in many plants, little is known about it in moso bamboo. In this present study, we identified 32 PheIDD family genes in moso bamboo and randomly sequenced the full-length open reading frames (ORFs) of ten PheIDDs. All PheIDDs shared a highly conserved IDD domain that contained two canonical C2H2-ZFs, two C2HC-ZFs, and a nuclear localization signal. Collinearity analysis showed that segmental duplication events played an important role in expansion of the PheIDD gene family. Synteny analysis indicated that 30 PheIDD genes were orthologous to those of rice (Oryza sativa). Thirty PheIDDs were expressed at low levels, and most PheIDDs exhibited characteristic organ-specific expression patterns. Despite their diverse expression patterns in response to exogenous plant hormones, 8 and 22 PheIDDs responded rapidly to IAA and 6-BA treatments, respectively. The expression levels of 23 PheIDDs were closely related to the outgrowth of aboveground branches and 20 PheIDDs were closely related to the awakening of underground dormant buds. In addition, we found that the PheIDD21 gene generated two products by alternative splicing. Both isoforms interacted with PheDELLA and PheSCL3. Furthermore, both isoforms could bind to the cis-elements of three genes (PH02Gene17121, PH02Gene35441, PH02Gene11386). Taken together, our work provides valuable information for studying the molecular breeding mechanism of lateral organ development in moso bamboo.
Structure of AQEE-30 of VGF Neuropeptide in Membrane-Mimicking Environments
Authors: Park One-Sung , Bang Jeong-Kyu , Cheong Chaejoon , Jeon Young-Ho
Published: November 2022

AQEE-30 is one of the VGF peptides, which are derived from the VGF polypeptide precursor, and related to various physiological phenomena including neuroprotective effects in Huntington′s disease and amyotrophic lateral sclerosis (ALS). Although various functions of AQEE-30 have been reported so far, the structure of this peptide has not been reported yet. In this study, the structure of human AQEE-30 was investigated in hexafluoroisopropanol (HFIP) and dodecyl phosphocholine (DPC) micelle solutions, using circular dichroism (CD) and nuclear magnetic resonance (NMR) spectroscopy. CD results showed that AQEE-30 had a partial helical structure in aqueous buffer, and the helical structure was stabilized in the HFIP and DPC micelle solutions. The 3D structures determined by NMR spectroscopy showed that AQEE-30 adopted mainly α-helical structure in both the HFIP and DPC micelle solutions. The surface of AQEE-30 showed that it was predominantly negatively charged. The residues from 601 to 611 in both the HFIP and DPC micelle solutions showed amphiphilicity with four negatively charged residues, glutamate. The C-terminal consecutive arginine residues formed a partial positively charged surface. These results suggest an α-helical active structure of AQEE-30 in the cell-membrane environment.
Generation of the First Human In Vitro Model for McArdle Disease Based on iPSC Technology
Authors: Ortuño-Costela María del Carmen , Cerrada Victoria , Moreno-Izquierdo Ana , García-Consuegra Inés , Laberthonnière Camille , Delourme Mégane , Garesse Rafael , Arenas Joaquín , Fuster García Carla , García García Gema , Millán José María , Magdinier Frédérique , Gallardo María Esther
Published: November 2022

McArdle disease is a rare autosomal recessive disorder caused by mutations in the PYGM gene. This gene encodes for the skeletal muscle isoform of glycogen phosphorylase (myophosphorylase), the first enzyme in glycogenolysis. Patients with this disorder are unable to obtain energy from their glycogen stored in skeletal muscle, prompting an exercise intolerance. Currently, there is no treatment for this disease, and the lack of suitable in vitro human models has prevented the search for therapies against it. In this article, we have established the first human iPSC-based model for McArdle disease. For the generation of this model, induced pluripotent stem cells (iPSCs) from a patient with McArdle disease (harbouring the homozygous mutation c.148C>T; p.R50* in the PYGM gene) were differentiated into myogenic cells able to contract spontaneously in the presence of motor neurons and generate calcium transients, a proof of their maturity and functionality. Additionally, an isogenic skeletal muscle model of McArdle disease was created. As a proof-of-concept, we have tested in this model the rescue of PYGM expression by two different read-through compounds (PTC124 and RTC13). The developed model will be very useful as a platform for testing drugs or compounds with potential pharmacological activity.
New Effective Method of Lactococcus Genome Editing Using Guide RNA-Directed Transposition
Authors: Pechenov Pavel Yu , Garagulya Danil A. , Stanovov Daniil S. , Letarov Andrey V.
Published: November 2022

Lactococcus lactis is an important industrial microorganism and a widely used model object for research in the field of lactic acid bacteria (LAB) biology. The development of new L. lactis and related LAB strains with improved properties, including phage-resistant strains for dairy fermentation, LAB-based vaccines or strains with altered genotypes for research purposes, are hindered by the lack of genome-editing tools that allow for the easy and straightforward incorporation of a significant amount of the novel genetic material, such as large genes or operons, into the chromosomes of these bacteria. We recently employed a suggested system based on the CRISPR–Cas-associated transposon for the editing of the L. lactis genome. After the in-depth redesign of the system, we were able to achieve the stable incorporation of the fragments that were sized up to 10 kbp into the L. lactis beta-galactosidase gene. The efficiency of editing under the optimized conditions were 2 × 10−4 and 4 × 10−5 for 1 kbp and 10 kbp, respectively, which are sufficient for fast and easy modifications if a positive selection marker can be used.
Towards Zebrafish Models of CNS Channelopathies
Authors: Kolesnikova Tatiana O. , Demin Konstantin A. , Costa Fabiano V. , Zabegalov Konstantin N. , de Abreu Murilo S. , Gerasimova Elena V. , Kalueff Allan V.
Published: November 2022

Channelopathies are a large group of systemic disorders whose pathogenesis is associated with dysfunctional ion channels. Aberrant transmembrane transport of K+, Na+, Ca2+ and Cl− by these channels in the brain induces central nervous system (CNS) channelopathies, most commonly including epilepsy, but also migraine, as well as various movement and psychiatric disorders. Animal models are a useful tool for studying pathogenesis of a wide range of brain disorders, including channelopathies. Complementing multiple well-established rodent models, the zebrafish (Danio rerio) has become a popular translational model organism for neurobiology, psychopharmacology and toxicology research, and for probing mechanisms underlying CNS pathogenesis. Here, we discuss current prospects and challenges of developing genetic, pharmacological and other experimental models of major CNS channelopathies based on zebrafish.