bluuebunny/crossref_metadata_2025 huggingface dataset:
From First to Second: How Stickler’s Diagnostic Genetics Has Evolved to Match Sequencing Technologies
Authors: Martin Howard , Richards Allan J. , Snead Martin P.
Published: June 2022

Diagnostic genetics within the United Kingdom National Health Service (NHS) has undergone many stepwise improvements in technology since the completion of the human genome project in 2003. Although Sanger sequencing has remained a cornerstone of the diagnostic sequencing arena, the human genome reference sequence has enabled next-generation sequencing (more accurately named ‘second-generation sequencing’), to rapidly surpass it in scale and potential. This mini review discusses such developments from the viewpoint of the Stickler’s higher specialist service, detailing the considerations and improvements to diagnostic sequencing implemented since 2003.
A Novel 90-kbp Deletion of RUNX2 Associated with Cleidocranial Dysplasia
Authors: Zhang Yanli , Duan Xiaohong
Published: June 2022

Cleidocranial dysplasia (CCD) is a rare autosomal dominant skeletal dysplasia caused by runt-related transcription factor 2 (RUNX2) mutations. In addition to the regular missense, small or large fragment deletions are the common mutation types of RUNX2. This study aimed to find the rules of deletions in RUNX2. The clinical information of one Chinese CCD family was collected. Genomic DNA was extracted for whole-exome sequencing (WES). Bioinformatics analyzed the pathogenicity of the variants. Polymerase chain reaction (PCR) and Sanger sequencing were carried out using specific primers. RT-PCR and Q-PCR were also used to detect the mRNA level of RUNX2. The CCD studies related with deletions in RUNX2 from 1999 to 2021 from HGMD and PubMed were collected and analyzed for the relationship between the phenotypes and the length of deleted fragments. The proband presented typical CCD features, including delayed closure of cranial sutures, clavicle dysplasia, abnormal teeth. WES, PCR with specific primers and Sanger sequencing revealed a novel heterozygous 90-kbp deletion in RUNX2 (NG_008020.2 g.103671~193943), which caused a substitution (p.Asn183Ile) and premature termination (p.Asp184*). In addition, the mRNA expression of RUNX2 was decreased by 75.5% in the proband. Herein, 31 types of deletions varying from 2 bp to 800 kbp or covering the whole gene of RUNX2 were compared and the significant phenotypic difference was not found among these deletions. The CCD phenotypes were related with the final effects of RUNX2 mutation instead of the length of deletion. WES has the defects in identifying large indels, and direct PCR with specific primers and Sanger sequencing could make up for the shortcoming.
Genotype-Phenotype Correlations in Neurofibromatosis Type 1: Identification of Novel and Recurrent NF1 Gene Variants and Correlations with Neurocognitive Phenotype
Authors: Napolitano Filomena , Dell’Aquila Milena , Terracciano Chiara , Franzese Giuseppina , Gentile Maria Teresa , Piluso Giulio , Santoro Claudia , Colavito Davide , Patanè Anna , De Blasiis Paolo , Sampaolo Simone , Paladino Simona , Melone Mariarosa Anna Beatrice
Published: June 2022

Neurofibromatosis type 1 (NF1) is one of the most common genetic tumor predisposition syndrome, caused by mutations in the NF1. To date, few genotype-phenotype correlations have been discerned in NF1, due to a highly variable clinical presentation. We aimed to study the molecular spectrum of NF1 and genotype-phenotype correlations in a monocentric study cohort of 85 NF1 patients (20 relatives, 65 sporadic cases). Clinical data were collected at the time of the mutation analysis and reviewed for accuracy in this investigation. An internal phenotypic categorization was applied. The 94% of the patients enrolled showed a severe phenotype with at least one systemic complication and a wide range of associated malignancies. Spine deformities were the most common complications in this cohort. We also reported 66 different NF1 mutations, of which 7 are novel mutations. Correlation analysis identified a slight significant inverse correlation between age at diagnosis and delayed acquisition of psychomotor skills with residual multi-domain cognitive impairment. Odds ratio with 95% confidence interval showed a higher prevalence of learning disabilities in patients carrying frameshift mutations. Overall, our results aim to offer an interesting contribution to studies on the genotype–phenotype of NF1 and in genetic management and counselling.
Evaluation of Expressive Arts Therapy on the Resilience of University Students in COVID-19: A Network Analysis Approach
Authors: Li Yue , Peng Jun
Published: June 2022

As an alternative to traditional verbal counselling, expressive arts therapy has been shown to be an effective method of mental health care, particularly when dealing with stressful public interactions, such as those associated with COVID-19. However, few studies have been conducted to determine the efficacy of expressive arts therapy on the resilience of psychologically exposed university students during COVID-19. Furthermore, since network analysis appears to be a popular approach in psychological research, it has not been used in recent intervention studies for resilience. As a result, the current study utilized a network analysis approach to determine the efficacy of expressive arts therapy on the resilience of university students during the COVID-19 pandemic. A total of 263 students in a comprehensive university in China were selected for the therapy group between March and November 2021. In a pre-post design, students’ resilience was assessed using the Resiliency Scale for University Students (RSUS). The extended Bayesian information criteria (EBIC) and graphical LASSO were used to estimate and define paired resilience networks, and the strength, betweenness, and closeness indices were utilized to determine the centrality of the six facets of resilience. Additionally, we verified the stability and accuracy. It was discovered that significant differences appeared between the paired networks before and after expressive arts therapy. Facets of self-efficacy, self-acceptance and problem-solving in resilience were notably improved after the therapy, with the variable of emotional stability sustained at the mean level. Meanwhile, the network analysis has highlighted the central variable of self-efficacy in the pre-intervention and support from friends in the post-intervention. The connectivity among the components of problem solving, support from friends, and support from family was enhanced, with support from friends playing the role of hub nod in the following network. By utilizing a network analytic approach, expressive arts therapy can be more targeted in intervening in resilience mechanisms. As a proxy for efficacious problem-solving, intervention should be calibrated to the cultivation of social support networks, especially in the support from friends.
Dietary Habits and Lifestyle Behaviors of Saudi Residents during the COVID-19 Pandemic: A Cross-Sectional Study
Authors: Bushnaq Taqwa , Algheshairy Reham M. , Almujaydil Mona S. , Malki Asma Al , Alharbi Hend F. , Barakat Hassan
Published: June 2022

Coronavirus disease 2019 (COVID-19) has continued to spread rapidly worldwide, forcing countries to enact lockdowns and strict social-distancing measures that affect individual and population health. This study investigates the changes in dietary habits and lifestyle behaviors among Saudi residents during the COVID-19 pandemic. A cross-sectional survey was conducted on 786 participants, with a mean age of 30 years. The questionnaire was administered online and sent via social media applications. Dietary habits, physical activity, TV-watching time, sleep patterns, stressors, and socio-demographic data were evaluated. Among the survey respondents, there was an increase in weight gain, food consumption, and decreased physical activity. Respondents who gained weight consumed more snacks, ate larger food portions, and engaged in less physical activity. Furthermore, a high fish, water, and supplements intake was noticed. Although respondents’ intake of fruits and vegetables was low, most individuals reported a negatively changed intake. However, most respondents reported increased consumption of lean meat and sweet foods. Observing changes in dietary and lifestyle habits during the lockdown period may help elaborate on the pandemic’s consequences for population health and assist in formulating and implementing policies for future closures, while supporting vulnerable groups.
The Impacts of Prenatal Mental Health Issues on Birth Outcomes during the COVID-19 Pandemic: A Scoping Review
Authors: Zhao Tianqi , Zuo Hanxiao , Campbell Sandra M. , Jhangri Gian S. , Dobson Keith S. , Li Jessica Yijia , Premji Shahirose S. , Tao Fangbiao , Zhu Beibei , Yamamoto Shelby S.
Published: June 2022

Background: The severity of the COVID-19 pandemic is likely to exacerbate mental health problems during the prenatal period and increase the risk of adverse birth outcomes. This review assessed the published literature related to the impacts of prenatal mental health issues on birth outcomes during the COVID-19 pandemic. Methods: This scoping review was conducted using PROSPERO, Cochrane Library, OVID Medline, Ovid EMBASE, OVID PsycInfo, EBSCO CINAHL, and SCOPUS. The search was conducted using controlled vocabulary and keywords representing the concepts “COVID19”, “mental health” and “birth outcomes”. The main inclusion criteria were peer-reviewed published articles from late 2019 to the end of July 2021. Results and Discussion: After removing duplicates, 642 articles were identified, of which two full texts were included for analysis. Both articles highlighted that pregnant women have experienced increasing prenatal mental health issues during the COVID-19 pandemic and, further, increased the risk of developing adverse births. This scoping review highlighted that there is a lack of research on the impact of prenatal mental health issues on birth outcomes during the pandemic. Conclusion: Given the severity of the COVID-19 pandemic and the burdens of prenatal mental health issues and adverse birth outcomes, there is an urgent need to conduct further research.
Kinematics of Cervical Spine during Rowing Ergometer at Different Stroke Rates in Young Rowers: A Pilot Study
Authors: Giustino Valerio , Zangla Daniele , Messina Giuseppe , Pajaujiene Simona , Feka Kaltrina , Battaglia Giuseppe , Bianco Antonino , Palma Antonio , Patti Antonino
Published: June 2022

Background: Research on biomechanics in rowing has mostly focused on the lumbar spine. However, injuries can also affect other body segments. Thus, the aim of this pilot study was to explore any potential variations in the kinematics of the cervical spine during two different stroke rates on the rowing ergometer in young rowers. Methods: Twelve young rowers of regional or national level were recruited for the study. The experimental protocol consisted of two separate test sessions (i.e., a sequence of 10 consecutive strokes for each test session) at different stroke rates (i.e., 20 and 30 strokes/min) on an indoor rowing ergometer. Kinematics of the cervical spine was assessed using an inertial sensor capable of measuring joint ROM (angle of flexion, angle of extension, total angle of flexion–extension). Results: Although there were no differences in the flexion and total flexion–extension movements between the test sessions, a significant increase in the extension movement was found at the highest stroke rate (p = 0.04, d = 0.66). Conclusion: Young rowers showed changes in cervical ROM according to stroke rate. The lower control of the head during the rowing stroke cycle can lead to a higher compensation resulting in an augmented effort, influencing sports performance, and increasing the risk of injury.
Feeding Practices, Maternal and Neonatal Outcomes in Vaginal Birth after Cesarean and Elective Repeat Cesarean Delivery
Authors: Rudzinski Patryk , Lopuszynska Inga , Pieniak Katarzyna , Stelmach Daria , Kacperczyk-Bartnik Joanna , Romejko-Wolniewicz Ewa
Published: June 2022

Cesarean section rates are constantly rising, and the number of women with a prior cesarean considering a delivery mode for their next labor is increasing. We aimed to compare maternal and neonatal outcomes and feeding method in women undergoing vaginal birth after cesarean (VBAC) versus elective repeat cesarean delivery (ERCD). This was a retrospective cohort study of women with one prior cesarean delivery (CD) and no previous vaginal births, delivering vaginally or by a CD in a single institution between 2016 and 2018. 355 live singleton spontaneous vaginal and cesarean deliveries were included. 121 women delivered vaginally and 234 had a CD. Neonates born by a CD were more likely to have higher birth weight (p < 0.001), higher weight at discharge (p < 0.001), macrosomia (p = 0.030), lose >10% of their body mass (p = 0.001), be mixed-fed (p < 0.001), and be hospitalized longer (p < 0.001). Children born vaginally were more likely to be exclusively breastfed (p < 0.001). Women undergoing VBAC were more likely to deliver preterm (p = 0.006) and post-term (p < 0.001), present with PROM (p < 0.001), have greater PROM latency period (p < 0.001), and experience intrahepatic cholestasis of pregnancy (p = 0.029), postpartum anemia (p < 0.001), and peripartum blood loss >1 L (p = 0.049). The incidence of anemia during pregnancy was higher in the ERCD cohort (p = 0.047). Women undergoing VBAC are more likely to breastfeed their children, perhaps for the same reason they choose the vaginal method of delivery, as vaginal delivery and breastfeeding along with antibiotic use, are the most important factors decreasing the risk for future diseases in their offspring.
The Effects of the Ecological Conservation Redline in China: A Case Study in Anji County
Authors: Zhang Chao , Lin Dayi , Wang Lixia , Hao Haiguang , Li Yuanyuan
Published: June 2022

The Ecological Conservation Redline (ECR) of China plays an important role in avoiding ecological space occupancy and maintaining regional ecological security. Anji County in Zhejiang Province is one of the first regions to implement the ECR in China. This paper takes Anji County as an example to analyze the effects of ECR. To do this, we first set up two scenarios with the CLUE-S model: a normal land-use development scenario (NLDS) and an ECR implementation scenario (ECRS); then we compare the land use of 2010 and 2015 under NLDS and ECRS. Land use, ecosystem services value (ESV), landscape metrics, and ecological product outputs were compared between the entire county and the ECR areas. The results revealed the following: (1) From 2000 to 2015, the ecological land in Anji County decreased by 4.03%, while it decreased by 1.17% in the ECR areas. (2) In the ECR areas, there was less arable land and construction land of the ECRS than in the NLDS, which indicates the ECR impeded the expansion of construction land and arable land in the ECR areas. (3) The ECR areas account for 39% of Anji County but contribute more than 80% to the ESV of the whole county. During 2000–2015, the ESV of the entire county decreased while the ESV of the ECR areas increased. (4) From 2000 to 2015, whereas landscape fragmentation of the entire county increased, that of ECR areas decreased. (5) Since the ECR’s implementation, Anji County has vigorously developed the bamboo industry, ecological agriculture, the tourism industry, and achieved rapid economic development via industrial restructuring and transformation. On the whole, the ECR has neither adversely affected land development nor economic development but instead has promoted the optimization of the land’s spatial development pattern.
Are Australian Universities Perpetuating the Teaching of Racism in Their Undergraduate Nurses in Discrete Aboriginal and Torres Strait Islander Courses? A Critical Race Document Analysis Protocol
Authors: Laccos-Barrett Keera , Brown Angela Elisabeth , West Roianne , Baldock Katherine Lorraine
Published: June 2022

Systemic racism has a profound negative impact on the health outcomes of Australia’s First Nations peoples, hereafter referred to as Aboriginal and Torres Strait Islander peoples, where racism and white privilege have largely become normalised and socially facilitated. A national framework is being mobilised within the tertiary-level nursing curriculum to equip future health professionals with cultural capabilities to ensure culturally safe, equitable health care for Aboriginal and Torres Strait Islander peoples. In 2019, nurses comprised more than half of all registered health professionals in Australia, and current national standards for nursing state that Australian universities should be graduating registered nurses capable of delivering care that is received as culturally safe. It is therefore critical to evaluate where learning objectives within nursing curricula may lead to the reinforcement and teaching of racist ideologies to nursing students. This protocol outlines a framework and methodology that will inform a critical race document analysis to evaluate how learning objectives assert the social construction of “race” as a tool of oppressive segregation. The document analysis will include each discrete Aboriginal and Torres Strait Islander health course within all undergraduate nursing programs at Australian universities. The approach outlined within this protocol is developed according to an Indigenous research paradigm and Colonial Critical Race Theory as both the framework and methodology. The purpose of the framework is a means for improving health professional curriculum by reducing racism as highlighted in nation-wide strategies for curriculum reform.
Tribal Tobacco Use Project II: Planning, Implementation, and Dissemination Using Culturally Relevant Data Collection among American Indian Communities
Authors: Roland Kendra M. , Anderson Madison D. , Carroll Dana M. , Webber Anna G. , Rhodes Kristine L. , Poupart John , Forster Jean L. , Peterson-Hickey Melanie , Pickner Wyatt J.
Published: June 2022

American Indians have substantially higher commercial tobacco-related cancer rates when compared to the general population. To effectively combat commercial tobacco-related cancer, it is important that tribal nations obtain current and accurate community-specific data on commercial tobacco use and exposure-related attitudes and behaviors. With the goal to collect, synthesize, and disseminate data on tobacco use, including the role traditional tobacco plays among American Indian people, the American Indian Cancer Foundation (AICAF) and various stakeholders developed and implemented the Tribal Tobacco Use Project II (TTUP II) during 2018–2021. Building upon its predecessor, the Tribal Tobacco Use Project I (TTUP I), TTUP II used principles of community-based participatory research and culturally appropriate methods, such as Reality-Based Research, in partnership with tribal nations. We describe the TTUP II rationale, methods for participant recruitment and data collection, emphasizing the importance of using culturally relevant survey items to disentangle commercial tobacco use from traditional tobacco use. American Indian traditional tobacco is viewed as medicine in these communities with a unique socio-cultural context that must be addressed when engaging in commercial tobacco control efforts in American Indian communities. This approach may be useful to other tribal nations who are interested in conducting culturally relevant tobacco surveillance efforts.
Family Alliance and Intergenerational Transmission of Coparenting in Gay and Heterosexual Single-Father Families through Surrogacy: Associations with Child Attachment Security
Authors: Carone Nicola
Published: June 2022

Parents tend to internalize the coparenting model they experienced during childhood and enact it in their coparenting relationships as adults. These interactive patterns may, in turn, shape their children’s internal working models of attachment relationships. The present study recruited 31 gay and 28 heterosexual single-father families through surrogacy to examine family alliance quality and the mediating role of observed supportive and conflictual coparenting in the association between the coparenting quality single fathers experienced in their families of origin and the attachment security of their children. All single fathers lived in Italy, were cisgender and White, and had a child aged 6–12 years (M = 97.73 months; SD = 20.48; 47.5% girls) who they coparented with nonparental caregivers (i.e., 33 grandparents, 18 babysitters, 8 uncles/aunts). Families did not differ in family alliance dimensions based on fathers’ sexual orientation. Additionally, single fathers who experienced greater coparenting quality in their families of origin demonstrated lower levels of conflictual coparenting, which, in turn, were associated with greater child attachment security. In contrast, observed supportive coparenting did not mediate this relation. The results emphasize the need to reconceptualize the dyadic coparental unit in single-father surrogacy families to include extended family members and nonrelatives.
Epidemiology of Δ8THC-Related Carcinogenesis in USA: A Panel Regression and Causal Inferential Study
Authors: Reece Albert Stuart , Hulse Gary Kenneth
Published: June 2022

The use of Δ8THC is increasing at present across the USA in association with widespread cannabis legalization and the common notion that it is “legal weed”. As genotoxic actions have been described for many cannabinoids, we studied the cancer epidemiology of Δ8THC. Data on 34 cancer types was from the Centers for Disease Control Atlanta Georgia, substance abuse data from the Substance Abuse and Mental Health Services Administration, ethnicity and income data from the U.S. Census Bureau, and cannabinoid concentration data from the Drug Enforcement Agency, were combined and processed in R. Eight cancers (corpus uteri, liver, gastric cardia, breast and post-menopausal breast, anorectum, pancreas, and thyroid) were related to Δ8THC exposure on bivariate testing, and 18 (additionally, stomach, Hodgkins, and Non-Hodgkins lymphomas, ovary, cervix uteri, gall bladder, oropharynx, bladder, lung, esophagus, colorectal cancer, and all cancers (excluding non-melanoma skin cancer)) demonstrated positive average marginal effects on fully adjusted inverse probability weighted interactive panel regression. Many minimum E-Values (mEVs) were infinite. p-values rose from 8.04 × 10−78. Marginal effect calculations revealed that 18 Δ8THC-related cancers are predicted to lead to a further 8.58 cases/100,000 compared to 7.93 for alcoholism and −8.48 for tobacco. Results indicate that between 8 and 20/34 cancer types were associated with Δ8THC exposure, with very high effect sizes (mEVs) and marginal effects after adjustment exceeding tobacco and alcohol, fulfilling the epidemiological criteria of causality and suggesting a cannabinoid class effect. The inclusion of pediatric leukemias and testicular cancer herein demonstrates heritable malignant teratogenesis.
Associative Analysis of Inefficiencies and Station Activity Levels in Emergency Response
Authors: Tiam-Lee Thomas James , Henriques Rui , Manquinho Vasco
Published: June 2022

Emergency medical services (EMS) around the world face the challenging task of allocating resources to efficiently respond to medical emergencies within a geographical area. While several studies have been done to improve various aspects of EMS, such as ambulance dispatch planning and station placement optimization, few works have focused on the assessment of existing rich real-world emergency response data to systematically identify areas of improvement. In this paper, we propose DAPI (data-driven analysis of potential response inefficiencies), a general tool for analyzing inefficiencies in emergency response datasets. DAPI efficiently identifies potential response bottlenecks based on spatial distributions of ambulance responses and statistically assesses them with respect to inferred activity levels of relevant dispatch stations to aid causality analysis. DAPI is applied on a dataset containing all medical emergency responses in mainland Portugal, in which we find statistical evidence that inefficiencies are correlated with high levels of activity of stations closer to an emergency location. We present these findings, along with the associated patterns and geographical clusters, serving as a valuable decision support tool to aid EMS in improving their operations.
An Augmented Geospatial Service Web Based on QoS Constraints and Geospatial Service Semantic Relationships
Authors: Jin Fengying , Li Rui , Liang Jianyuan , Zhang Xianyuan , Xing Huaqiao , Gui Zhipeng , Wu Huayi
Published: June 2022

The service network is capable of addressing large-scale service composition. However, existing service network works still have several limitations. Prior knowledge, such as expert-defined service chains, is not incorporated into the service network. QoS constraints are less considered in the service network, and thus the generated service chain does not always satisfy the optimal QoS constraints. Additionally, some basic services also require outputs to be used directly as inputs, which the service network cannot provide. To address these limitations, this paper proposes a geospatial service web (GSW) model named SR-QoS-GSW that incorporates service semantic relationships and QoS information. The SR-QoS-GSW model consists of atomic services and composite services that consider QoS, processing services, data services, and relationships among them. A SR-QoS-GSW prototype was developed using 570 atomic services and 27 composite services and evaluated using two case studies—a river network extraction and an urban housing selection. Then, the information entropy and time complexity between SR-QoS-GSW and the existing service network were compared. The results show that geospatial service chains can be created more efficiently by incorporating existing service chains as composite services. Integrating QoS information into the GSW would allow service composition algorithms to generate service chains that satisfy optimal QoS constraints. The outputs of services used as new inputs with additional self-matching relationships also give the service network greater flexibility. Finally, the analysis of the information entropy and time complexity verified the increased diversity and decreased the search space of the SR-QoS-GSW.
Extraction of Continuous and Discrete Spatial Heterogeneities: Fusion Model of Spatially Varying Coefficient Model and Sparse Modelling
Authors: Inoue Ryo , Den Koichiro
Published: June 2022

Geospatial phenomena often have spatial heterogeneity, which is caused by differences in the data generation process from place to place. There are two types of spatial heterogeneity: continuous and discrete, and there has been much discussion about how to analyze one type of spatial heterogeneity. Although geospatial phenomena can have both types of spatial heterogeneities, previous studies have not sufficiently discussed how to consider these two different types of spatial heterogeneity simultaneously and how to detect them separately, which may lead to biased estimates and the wrong interpretation of geospatial phenomena. This study proposes a new approach for the analysis of spatial data with both heterogeneities by combining the eigenvector spatial filtering-based spatially varying coefficient (ESF-SVC) model, which assumes the continuous spatial heterogeneity and generalized lasso (GL) estimation, which assumes discrete spatial heterogeneity and proposes the ESF-GL-SVC model. The performance of ESF-GL-SVC was evaluated through experiments based on a Monte Carlo simulation and confirms that the ESF-GL-SVC showed better performance in estimating coefficients with both types of spatial heterogeneity than the previous two models. The application of the apartment rent data showed that the ESF-GL-SVC outputs the result with the smallest BIC value, and the estimated coefficients depict continuous and discrete spatial heterogeneity in the dataset. Reasonable coefficients were estimated using the ESF-GL-SVC, although some coefficients by ESF-SVC were not.
Data-Driven Identification of Biomarkers for In Situ Monitoring of Drug Treatment in Bladder Cancer Organoids
Authors: Becker Lucas , Fischer Felix , Fleck Julia L. , Harland Niklas , Herkommer Alois , Stenzl Arnulf , Aicher Wilhelm K. , Schenke-Layland Katja , Marzi Julia
Published: June 2022

Three-dimensional (3D) organoid culture recapitulating patient-specific histopathological and molecular diversity offers great promise for precision medicine in cancer. In this study, we established label-free imaging procedures, including Raman microspectroscopy (RMS) and fluorescence lifetime imaging microscopy (FLIM), for in situ cellular analysis and metabolic monitoring of drug treatment efficacy. Primary tumor and urine specimens were utilized to generate bladder cancer organoids, which were further treated with various concentrations of pharmaceutical agents relevant for the treatment of bladder cancer (i.e., cisplatin, venetoclax). Direct cellular response upon drug treatment was monitored by RMS. Raman spectra of treated and untreated bladder cancer organoids were compared using multivariate data analysis to monitor the impact of drugs on subcellular structures such as nuclei and mitochondria based on shifts and intensity changes of specific molecular vibrations. The effects of different drugs on cell metabolism were assessed by the local autofluorophore environment of NADH and FAD, determined by multiexponential fitting of lifetime decays. Data-driven neural network and data validation analyses (k-means clustering) were performed to retrieve additional and non-biased biomarkers for the classification of drug-specific responsiveness. Together, FLIM and RMS allowed for non-invasive and molecular-sensitive monitoring of tumor-drug interactions, providing the potential to determine and optimize patient-specific treatment efficacy.
Lmx1a-Dependent Activation of miR-204/211 Controls the Timing of Nurr1-Mediated Dopaminergic Differentiation
Authors: Pulcrano Salvatore , De Gregorio Roberto , De Sanctis Claudia , Lahti Laura , Perrone-Capano Carla , Ponti Donatella , di Porzio Umberto , Perlmann Thomas , Caiazzo Massimiliano , Volpicelli Floriana , Bellenchi Gian Carlo
Published: June 2022

The development of midbrain dopaminergic (DA) neurons requires a fine temporal and spatial regulation of a very specific gene expression program. Here, we report that during mouse brain development, the microRNA (miR-) 204/211 is present at a high level in a subset of DA precursors expressing the transcription factor Lmx1a, an early determinant for DA-commitment, but not in more mature neurons expressing Th or Pitx3. By combining different in vitro model systems of DA differentiation, we show that the levels of Lmx1a influence the expression of miR-204/211. Using published transcriptomic data, we found a significant enrichment of miR-204/211 target genes in midbrain dopaminergic neurons where Lmx1a was selectively deleted at embryonic stages. We further demonstrated that miR-204/211 controls the timing of the DA differentiation by directly downregulating the expression of Nurr1, a late DA differentiation master gene. Thus, our data indicate the Lmx1a-miR-204/211-Nurr1 axis as a key component in the cascade of events that ultimately lead to mature midbrain dopaminergic neurons differentiation and point to miR-204/211 as the molecular switch regulating the timing of Nurr1 expression.
Silicon Controls Bacterial Wilt Disease in Tomato Plants and Inhibits the Virulence-Related Gene Expression of Ralstonia solanacearum
Authors: Wang Lei , Gao Yang , Jiang Nihao , Yan Jian , Lin Weipeng , Cai Kunzheng
Published: June 2022

Silicon (Si) has a multifunctional role in improving plant growth and enhancing plant disease resistance, but its mechanisms are not fully understood. In this study, we investigated the impacts of silicon application on the control of bacterial wilt and elucidated the molecular mechanisms using transcriptome sequencing. Compared to non-Si treatment, Si application (0.5–2 mM) significantly reduces tomato bacterial wilt index by 46.31–72.23%. However, Si does not influence the growth of R. solanacearum. Si application negatively influences R. solanacearum exopolysaccharide (EPS) synthesis and biofilm formation. Transcriptome analysis showed that Si treatment significantly downregulates the expression of virulence genes’ transcriptional regulator (xpsR), EPS synthesis-related genes (epsD and tek), and type III effectors (HrpB2, SpaO, and EscR) in R. solanacearum. In addition, Si remarkably upregulates the expression of twitch motor-related genes (pilE2, pilE, fimT, and PilX). These findings suggest that silicon-suppressed tomato wilt incidence may be due to the regulation of the virulence-related genes of R. solanacearum by Si. Our research adds new knowledge to the application of Si in the field of disease control.
Friction in Myocardial Anoxia Leads to Negative Excess Entropy Production, Self-Organization, and Dissipative Structures
Authors: Lecarpentier Yves , Claes Victor , Hébert Jean-Louis , Krokidis Xénophon , Schussler Olivier , Vallée Alexandre
Published: June 2022

Contraction of the heart is caused by actin filaments sliding along myosin filaments. This generates a frictional force inducing wear of the contractile apparatus. We postulated that this process could be exacerbated when the heart was submitted to severe anoxia. Anoxia induced dramatic abnormalities in the molecular properties of actin-myosin crossbridges. We applied the formalism of far-from-equilibrium thermodynamics to the left ventricular papillary muscles (LVPMs) of mammalian rat hearts which had been subjected to a prolonged anoxia (3 h). We showed that when subjected to prolonged anoxia, the heart operated far-from-equilibrium as evidenced by the non-linearity between thermodynamic force (F/T: Frictional force/Kelvin temperature) and thermodynamic flow (v0: myofilament sliding velocity). The rate of entropy production (EPR) was the product of (F/T) and v0. The excess entropy production (EEP) was equal to ∂δ2S∂t = ∂FTδvo; (S: entropy). The tribological system remained stable when EEP was positive and became unstable when EEP became negative, thus characterizing instability of the system and reflecting the occurrence of self-organization and possibly dissipative structures. After 3 h anoxia, re-oxygenation induced significant reversibility. About 20% of the myosin heads did not recover despite re-oxygenation. These results may be of importance in the context of heart transplantation where the delay between the time of sampling from the donor and the time of the graft installation in the recipient should be as short as possible.
Variation of Structural and Dynamical Flexibility of Myelin Basic Protein in Response to Guanidinium Chloride
Authors: Haris Luman , Biehl Ralf , Dulle Martin , Radulescu Aurel , Holderer Olaf , Hoffmann Ingo , Stadler Andreas M.
Published: June 2022

Myelin basic protein (MBP) is intrinsically disordered in solution and is considered as a conformationally flexible biomacromolecule. Here, we present a study on perturbation of MBP structure and dynamics by the denaturant guanidinium chloride (GndCl) using small-angle scattering and neutron spin–echo spectroscopy (NSE). A concentration of 0.2 M GndCl causes charge screening in MBP resulting in a compact, but still disordered protein conformation, while GndCl concentrations above 1 M lead to structural expansion and swelling of MBP. NSE data of MBP were analyzed using the Zimm model with internal friction (ZIF) and normal mode (NM) analysis. A significant contribution of internal friction was found in compact states of MBP that approaches a non-vanishing internal friction relaxation time of approximately 40 ns at high GndCl concentrations. NM analysis demonstrates that the relaxation rates of internal modes of MBP remain unaffected by GndCl, while structural expansion due to GndCl results in increased amplitudes of internal motions. Within the model of the Brownian oscillator our observations can be rationalized by a loss of friction within the protein due to structural expansion. Our study highlights the intimate coupling of structural and dynamical plasticity of MBP, and its fundamental difference to the behavior of ideal polymers in solution.
Experimental and Theoretical Study of N2 Adsorption on Hydrogenated Y2C4H− and Dehydrogenated Y2C4− Cluster Anions at Room Temperature
Authors: Gao Min , Ding Yong-Qi , Ma Jia-Bi
Published: June 2022

The adsorption of atmospheric dinitrogen (N2) on transition metal sites is an important topic in chemistry, which is regarded as the prerequisite for the activation of robust N≡N bonds in biological and industrial fields. Metal hydride bonds play an important part in the adsorption of N2, while the role of hydrogen has not been comprehensively studied. Herein, we report the N2 adsorption on the well-defined Y2C4H0,1− cluster anions under mild conditions by using mass spectrometry and density functional theory calculations. The mass spectrometry results reveal that the reactivity of N2 adsorption on Y2C4H− is 50 times higher than that on Y2C4− clusters. Further analysis reveals the important role of the H atom: (1) the presence of the H atom modifies the charge distribution of the Y2C4H− anion; (2) the approach of N2 to Y2C4H− is more favorable kinetically compared to that to Y2C4−; and (3) a natural charge analysis shows that two Y atoms and one Y atom are the major electron donors in the Y2C4− and Y2C4H− anion clusters, respectively. This work provides new clues to the rational design of TM-based catalysts by efficiently doping hydrogen atoms to modulate the reactivity towards N2.
Immunorthodontics: Role of HIF-1α in the Regulation of (Peptidoglycan-Induced) PD-L1 Expression in Cementoblasts under Compressive Force
Authors: Yong Jiawen , Gröger Sabine , Meyle Joerg , Ruf Sabine
Published: June 2022

Patients with periodontitis undergoing orthodontic therapy may suffer from undesired dental root resorption. The purpose of this in vitro study was to investigate the molecular mechanisms resulting in PD-L1 expression of cementoblasts in response to infection with Porphyromonas gingivalis (P. gingivalis) peptidoglycan (PGN) and compressive force (CF), and its interaction with hypoxia-inducible factor (HIF)-1α molecule: The cementoblast (OCCM-30) cells were kinetically infected with various concentrations of P. gingivalis PGN in the presence and absence of CF. Western blotting and RT-qPCR were performed to examine the protein expression of PD-L1 and HIF-1α as well as their gene expression. Immunofluorescence was applied to visualize the localization of these proteins within cells. An HIF-1α inhibitor was added for further investigation of necroptosis by flow cytometry analysis. Releases of soluble GAS-6 were measured by ELISA. P. gingivalis PGN dose dependently stimulated PD-L1 upregulation in cementoblasts at protein and mRNA levels. CF combined with P. gingivalis PGN had synergistic effects on the induction of PD-L1. Blockade of HIF-1α inhibited the P. gingivalis PGN-inducible PD-L1 protein expression under compression, indicating an HIF-1α dependent regulation of PD-L1 induction. Concomitantly, an HIF-1α inhibitor decreased the GAS-6 release in the presence of CF and P. gingivalis PGN co-stimulation. The data suggest that PGN of P. gingivalis participates in PD-L1 up-regulation in cementoblasts. Additionally, the influence of compressive force on P. gingivalis PGN-induced PD-L1 expression occurs in HIF-1α dependently. In this regard, HIF-1α may play roles in the immune response of cementoblasts via immune-inhibitory PD-L1. Our results underline the importance of molecular mechanisms involved in bacteria-induced periodontics and root resorption.
Combined Therapy Using Human Corneal Stromal Stem Cells and Quiescent Keratocytes to Prevent Corneal Scarring after Injury
Authors: Jhanji Vishal , Santra Mithun , Riau Andri K. , Geary Moira L. , Yang Tianbing , Rubin Elizabeth , Yusoff Nur Zahirah Binte M. , Dhaliwal Deepinder K. , Mehta Jodhbir S. , Yam Gary Hin-Fai
Published: June 2022

Corneal blindness due to scarring is conventionally treated by corneal transplantation, but the shortage of donor materials has been a major issue affecting the global success of treatment. Pre-clinical and clinical studies have shown that cell-based therapies using either corneal stromal stem cells (CSSC) or corneal stromal keratocytes (CSK) suppress corneal scarring at lower levels. Further treatments or strategies are required to improve the treatment efficacy. This study examined a combined cell-based treatment using CSSC and CSK in a mouse model of anterior stromal injury. We hypothesize that the immuno-regulatory nature of CSSC is effective to control tissue inflammation and delay the onset of fibrosis, and a subsequent intrastromal CSK treatment deposited collagens and stromal specific proteoglycans to recover a native stromal matrix. Using optimized cell doses, our results showed that the effect of CSSC treatment for suppressing corneal opacities was augmented by an additional intrastromal CSK injection, resulting in better corneal clarity. These in vivo effects were substantiated by a further downregulated expression of stromal fibrosis genes and the restoration of stromal fibrillar organization and regularity. Hence, a combined treatment of CSSC and CSK could achieve a higher clinical efficacy and restore corneal transparency, when compared to a single CSSC treatment.
Immunohistochemical Profile of Parathyroid Tumours: A Comprehensive Review
Authors: Uljanovs Romans , Sinkarevs Stanislavs , Strumfs Boriss , Vidusa Liga , Merkurjeva Kristine , Strumfa Ilze
Published: June 2022

Immunohistochemistry remains an indispensable tool in diagnostic surgical pathology. In parathyroid tumours, it has four main applications: to detect (1) loss of parafibromin; (2) other manifestations of an aberrant immunophenotype hinting towards carcinoma; (3) histogenesis of a neck mass and (4) pathogenetic events, including features of tumour microenvironment and immune landscape. Parafibromin stain is mandatory to identify the new entity of parafibromin-deficient parathyroid neoplasm, defined in the WHO classification (2022). Loss of parafibromin indicates a greater probability of malignant course and should trigger the search for inherited or somatic CDC73 mutations. Aberrant immunophenotype is characterised by a set of markers that are lost (parafibromin), down-regulated (e.g., APC protein, p27 protein, calcium-sensing receptor) or up-regulated (e.g., proliferation activity by Ki-67 exceeding 5%) in parathyroid carcinoma compared to benign parathyroid disease. Aberrant immunophenotype is not the final proof of malignancy but should prompt the search for the definitive criteria for carcinoma. Histogenetic studies can be necessary for differential diagnosis between thyroid vs. parathyroid origin of cervical or intrathyroidal mass; detection of parathyroid hormone (PTH), chromogranin A, TTF-1, calcitonin or CD56 can be helpful. Finally, immunohistochemistry is useful in pathogenetic studies due to its ability to highlight both the presence and the tissue location of certain proteins. The main markers and challenges (technological variations, heterogeneity) are discussed here in the light of the current WHO classification (2022) of parathyroid tumours.
Interaction Analysis between the Arabidopsis Transcription Repressor VAL1 and Transcription Coregulators SIN3-LIKEs (SNLs)
Authors: Chen Chuanyou , Gong Xia , Li Yan , Li Haitao , Zhang Haitao , Liu Li , Liang Dacheng , Yuan Wenya
Published: June 2022

VIVIPAROUS1/ABSCISIC ACID INSENSITIVE3-LIKE1 (VAL1) encodes a DNA-binding B3 domain protein and plays essential roles in seed maturation and flowering transition by repressing genes through epigenetic silencing in Arabidopsis. SWI-INDEPENDENT3 (SIN3)-LIKEs (SNLs), which encode scaffold proteins for the assembly of histone deacetylase complexes and have six SIN3 homologues (SNL1–SNL6) in Arabidopsis thaliana, directly repress gene expression to regulate seed maturation and flowering transition. However, it remains unclear whether VAL1 and SNLs work together in repressing the expression of related genes. In this study, yeast two-hybrid and firefly luciferase complementation imaging assays revealed that VAL1 interacts with SNLs, which can be attributed to its own zinc-finger CW (conserved Cys (C) and Trp (W) residues) domain and the PAH (Paired Amphipathic Helices) domains of SNLs. Furthermore, pull-down experiments confirmed that the CW domain of VAL1 interacts with both intact protein and the PAH domains of SNLs proteins, and the co-immunoprecipitation assays also confirmed the interaction between VAL1 and SNLs. In addition, quantitative real-time PCR (qRT-PCR) analysis showed that VAL1 and SNLs were expressed in seedlings, and transient expression assays showed that VAL1 and SNLs were localized in the nucleus. Considered together, these results reveal that VAL1 physically interacts with SNLs both in vitro and in vivo, and suggest that VAL1 and SNLs may work together to repress the expression of genes related to seed maturation and flowering transition in Arabidopsis.
Biocontrol Potential of Endophytic Plant-Growth-Promoting Bacteria against Phytopathogenic Viruses: Molecular Interaction with the Host Plant and Comparison with Chitosan
Authors: Shahzad Gul-i-Rayna , Passera Alessandro , Maldera Giusva , Casati Paola , Marcello Iriti , Bianco Piero Attilio
Published: June 2022

Endophytic plant-growth-promoting bacteria (ePGPB) are interesting tools for pest management strategies. However, the molecular interactions underlying specific biocontrol effects, particularly against phytopathogenic viruses, remain unexplored. Herein, we investigated the antiviral effects and triggers of induced systemic resistance mediated by four ePGPB (Paraburkholderia fungorum strain R8, Paenibacillus pasadenensis strain R16, Pantoea agglomerans strain 255-7, and Pseudomonas syringae strain 260-02) against four viruses (Cymbidium Ring Spot Virus—CymRSV; Cucumber Mosaic Virus—CMV; Potato Virus X—PVX; and Potato Virus Y—PVY) on Nicotiana benthamiana plants under controlled conditions and compared them with a chitosan-based resistance inducer product. Our studies indicated that ePGPB- and chitosan-treated plants presented well-defined biocontrol efficacy against CymRSV and CMV, unlike PVX and PVY. They exhibited significant reductions in symptom severity while promoting plant height compared to nontreated, virus-infected controls. However, these phenotypic traits showed no association with relative virus quantification. Moreover, the tested defense-related genes (Enhanced Disease Susceptibility-1 (EDS1), Non-expressor of Pathogenesis-related genes-1 (NPR1), and Pathogenesis-related protein-2B (PR2B)) implied the involvement of a salicylic-acid-related defense pathway triggered by EDS1 gene upregulation.