bluuebunny/crossref_metadata_2025 huggingface dataset:
The curvilinear relationship between team member boundary‐spanning differentiation and creativity
Authors: Yao Jin , Feng Jie , Liu Xinmei
Published: February 2022

Team creativity, as one of the most important sources of organizational competitiveness and continuous innovation, requires rich and non‐redundant information from both internal and external sources. Although an emerging line of research has shed light on the role of team boundary‐spanning activity in promoting creativity, it has solely focused on the mean level of team boundary‐spanning activity and rarely addressed the fact that not all team members engage in boundary‐spanning behaviors to the same extent. In this study, we broaden the existing knowledge by examining when and how the pattern of team boundary spanning, particularly the differentiation in team member boundary‐spanning activities, promotes team creativity. Based on the categorization‐elaboration model, we propose a curvilinear relationship between team member boundary‐spanning differentiation and creativity that is mediated through information sharing. Further, we add an important layer of understanding to the nature of this curvilinear relationship by identifying team goal interdependence as an important moderator. Using multisource data collected from two time points in 70 teams, we have received general support for our hypotheses. As such, we enrich the conversation about the importance of team boundary spanning by answering “Whether team boundary‐spanning differentiation fosters creativity, and when and how?”
Winter anticyclone activities in Siberia and their relationship to the regional temperature anomaly
Authors: Lu Chuhan , Zhang Yumin , Guan Zhaoyong
Published: February 2022

Under global warming, the frequent occurrence of Eurasian extreme cold events in the Northern Hemisphere in recent years has attracted widespread attention. Winter Eurasia near‐surface anticyclone activities are closely related to low temperatures. This study uses a new objective anticyclone identification algorithm based on a Mask region‐based convolutional neural network (Mask R‐CNN) to detect long‐term winter Eurasian anticyclone activities. We found that most tracks of Eurasian anticyclones (30.5%) pass Siberia in winter. Anticyclone tracks over Siberia can be well separated into the eastward moving type, the northwest–southeast type, and the local type, with distinct associated circulation patterns. The energy dispersions of upstream Rossby waves have an important influence on the movement, maintenance and intensification of the three types. In association with anticyclones intensifying in Siberia, surface air temperatures in the mid‐latitudes of Eurasia, especially Siberia, are significantly lower. In particular, the local type is the most active (56.1%) and is significantly correlated with the winter temperature anomaly in most areas of East Asia. Further research indicates that the preceding autumn sea ice condition in the Barents Sea can be used as a precursor factor to modulate the local type anticyclone activity and winter temperature anomaly in East Asia. In years of less ice, a quasi‐meridional wave activity flux originating from the Barents Sea is more active, and the centre SLP of the local type anticyclone is higher with more cold air mass carried, which in turn results in colder weather in Siberia, the Mongolian Plateau and East Asia.
Surgical timing after preoperative chemotherapy is associated with oncologic outcomes in resectable colorectal liver metastases
Authors: Sutton Thomas L. , Wong Liam H. , Walker Brett S. , Dewey Elizabeth N. , Eil Robert L. , Ibewuike Uchechukwu , Chen Emerson Y. , Rocha Flavio G. , Billingsley Kevin G. , Mayo Skye C.
Published: February 2022

Introduction

Preoperative chemotherapy (POC) is often employed for patients with resectable colorectal liver metastasis (CRLM). The time to resection (TTR) following the end of chemotherapy may impact oncologic outcomes; this phenomenon has not been studied in CRLM.

Methods

We queried our institutional cancer database for patients with resected CRLM after POC from 2003 to 2019. TTR was calculated from date of last cytotoxic chemotherapy. Kaplan–Meier analysis and multivariable Cox proportional hazards modeling were used to analyze recurrence‐free survival (RFS) and overall survival (OS).

Results

We identified n = 187 patients. One hundred twenty‐four (66%) patients had a TTR of <2 months, while 63 (33%) had a TTR of ≥2 months. Median follow‐up was 36 months. On Kaplan–Meier analysis, patients with TTR ≥ 2 months had shorter RFS (median 11 vs. 17 months, p = 0.002) and OS (median 44 vs. 62 months, p < 0.001). On multivariable analysis, TTR ≥ 2 months was independently associated with worse RFS (hazard ratio [HR] = 1.54, 95% confidence interval [CI] = 1.06–2.22, p = 0.02) and OS (HR = 1.75, 95% CI = 1.11–2.77, p = 0.01).

Conclusion

TTR ≥ 2 months following POC is independently associated with worse oncologic outcomes in patients with resectable CRLM. We therefore recommend consideration for hepatic resection of CRLM within this window whenever feasible.
Strategies for efficient estimation of soil organic content at the local scale based on a national spectral database
Authors: Li Hongyi , Li Yuheng , Yang Meihua , Chen Songchao , Shi Zhou
Published: February 2022

Soil function degradation threatens the sustainable management of soil resources and soil organic matter (SOM) is a vital and important factor. Powerful measuring tools will become very important, especially in areas where data are poor or absent. The archive: China Soil Visible and Near Infrared (vis–NIR) Spectroscopy Library (CSSL) could help providea solution for less costly and fast measuring of SOM. The aim of this article was to compare SOM prediction performance according to three strategies: i) general global partial least squares regression (PLSR) using CSSL with and without spiking samples; ii) memory‐based learning (MBL) using CSSL with and without spiking samples; and iii) general PLSR using only spiking samples to predict soil organic matter in the target area. When using spiked subsets, we also investigated the prediction performance of the extra‐weighted (several copies) subsets. A series of spiking subsets were randomly selected from the total spiking samples, which were selected by conditioned Latin hypercube sampling (cLHS) from the target sites. We calculated only the mean squared Euclidean distance (msd) between the estimates density function (pds) of the principal components (PCs) of vis–NIR spectroscopy from the validation dataset and spiking subsets and statistically inferred the optimal sampling set size to be 30. Our study showed that global PLSR using CSSL spiked with the statistically optimal local samples can achieve higher predicted performance [with a mean root mean square error (RMSE) of 5.75]. MBL spiked with five extra‐weighted optimal spiking samples achieved the best accuracy with an RMSE of 3.98, an R2 of 0.70, a bias of 0.04, and an LCCC of 0.81. The msd is a simple and effective method to determine an adequate spiking set size using only vis–NIR data. These accurate predictions demonstrated the usefulness of statistically representative spiking and MBL for advanced large soil spectral libraries for SOM determination, which is currently lacking at large soil spectral libraries in use.
Simazine degradation in agroecosystems: Will it be affected by the type and amount of microplastic pollution?
Authors: Zhou Jie , Wen Yuan , Cheng Hongguang , Zang Huadong , Jones Davey L.
Published: February 2022

Plastics and herbicides represent two of the most extensive and persistent anthropogenic contaminants entering agroecosystems. The synergistic interaction of these pollutants on soil health, however, remains poorly understood. For the first time, we investigated the behavior of a common selective triazine herbicide (simazine) in soil containing 0%, 1%, 5%, 10%, and 20% (w/w) of two commonly found microplastics, namely polyethylene (PE) and polyvinyl chloride (PVC). The mineralization of 14C‐labeled simazine in soil decreased with the presence of both PE and PVC as indicated by the lower total 14CO2 loss. Consequently, the half‐life of simazine increased in the presence of microplastics, although there was little difference between microplastic types. The ratio of fungi‐to‐bacteria in the soil increased by 9%–18% after 1% of microplastics addition, while enzyme activities involved in C‐cycling decreased by 20%–46% in soil amended with PVC relative to the control. Further, enzyme activities were negatively correlated with the half‐life of simazine when added with PVC. Therefore, we ascribe the repression in simazine degradation to changes in microbial community structure (increased fungi‐to‐bacteria ratio) and reduced enzyme activities. Overall, microplastics accumulation in soil decreases herbicide breakdown resulting in herbicide residuals remaining, which in turn, may increase their risk of reaching ground or surface waters.
Local winds and encroaching currents drive summertime subsurface blooms over a narrow shelf
Authors: Schlosser Tamara L. , Lucas Andrew J. , Jones Nicole L. , Nash Jonathan D. , Ivey Gregory N.
Published: February 2022

The ability to forecast the biological productivity of the coastal ocean relies on the quantification of the physical processes that deliver nutrients to the euphotic zone. Here we explore these pathways using observations of the coupled biological and physical variability of waters offshore of the east coast of Tasmania in the summertime. The observations include an array of moored autonomous profilers deployed over an 18‐d period—providing continuous, full‐depth measurements of turbulent microstructure, temperature, velocity, and chlorophyll a (Chl a) fluorescence, complemented by shipboard nutrient measurements. Local upwelling was driven by the encroaching East Australian Current (EAC) extension onto the shelf and to a lesser extent the local winds. The interaction of the local winds and the encroaching boundary current was reflected in the shelf nutrient budget and led to a rapid increase in subsurface Chl a. Diffusive vertical fluxes had minimal impact on subsurface Chl a in the mid‐shelf and outer‐shelf. Upwelling‐favorable winds were too weak to drive significant vertical mixing, and mixing associated with the current‐driven Ekman transport was too deep compared to the euphotic zone depth. The observed subsurface Chl a did not reflect the satellite estimates of productivity. Since the EAC extension transports warm, low‐nutrient surface waters from the subtropics, satellite chlorophyll measurements decreased during the same period the depth‐averaged Chl a increased. This seeming paradox illustrated how long duration, full water column sampling can elucidate the coupled biological and physical processes that aid our ongoing effort to forecast the biological state of the coastal ocean.
Simultaneous Synthesis and Self‐Assembly of Bottlebrush Block Copolymers at Room Temperature via Photoinitiated RAFT Dispersion Polymerization
Authors: Liu Dongdong , Yang Shuaiqi , Peng Shaojie , Chen Ying , Zhang Li , Tan Jianbo
Published: February 2022

Bottlebrush polymers exhibiting unique properties have attracted considerable attention for applications in many research areas. Herein, the first simultaneous synthesis and self‐assembly of bottlebrush block copolymers at room temperature via photoinitiated polymerization‐induced self‐assembly (photo‐PISA) using multifunctional macromolecular chain transfer agents (macro‐CTAs) is reported. Comparing with linear block copolymers, the bottlebrush block copolymers can promote the formation of higher‐order morphologies (e.g., vesicles) when targeting similar degrees of polymerization (DPs). Moreover, a higher polymerization rate is observed in the case of bottlebrush block copolymers. Gel permeation chromatography (GPC) analysis shows that good polymerization control is maintained when synthesizing bottlebrush block copolymers by photo‐PISA. Finally, the obtained bottlebrush block copolymer vesicles are used as seeds for further chain extension and multicompartment nanoparticles with a sponge internal structure are formed. It is expected that this study will not only expand polymer architectures employed in PISA, but also provide a new strategy to synthesize polymer nanoparticles with unique structures.
A novel FAM83H variant causes familial amelogenesis imperfecta with incomplete penetrance
Authors: Bai Rui‐Qi , He Wen‐Bin , Peng Qian , Shen Su‐Hui , Yu Qian‐Qian , Du Juan , Tan Yue‐Qiu , Wang Yue‐Hong , Liu Bin‐Jie
Published: February 2022

Background

Amelogenesis imperfecta (AI) is known to be a monogenic genetic disease caused by a variety of genes demonstrating a wide spectrum of penetrance. FAM83H is reported to be involved in AI: however, whether FAM83H causes AI with incomplete penetrance is unclear.

Methods

Whole‐exome sequencing was performed on two patients with AI, and putative disease‐related variants were validated by Sanger sequencing. Bioinformatic and in vitro functional analyses were performed to functionally characterize the identified disease‐causing variants.

Results

We identified a novel heterozygous nonsense variant of FAM83H (NM_198488: c.1975G > T, p.Glu659Ter); in vitro functional analysis showed that this mutant produced mislocalized proteins and was deleterious. Surprisingly, the clinical manifestations of each of the six individuals carrying this variant were different, with one carrier appearing to be completely asymptomatic for AI.

Conclusion

Our findings expand the variant spectrum for FAM83H and the phenotypic spectrum for FAM83H‐associated AI and suggest that FAM83H‐mediated AI exhibits incomplete penetrance.
The Carter Center launches MH parity collaborative
Authors: Canady Valerie A.
Published: February 2022

The Carter Center on Feb. 23 announced the launch of the Mental Health Parity Collaborative, a new multistate newsroom partnership focused on exploring access to and inequities in mental health care in the United States, a news release stated. The Rosalynn Carter Center Mental Health Fellowships Program is partnering with the Center for Public Integrity (CPI) to advance coverage of access to mental health care, officials stated.
Automatic inverse treatment planning of Gamma Knife radiosurgery via deep reinforcement learning
Authors: Liu Yingzi , Shen Chenyang , Wang Tonghe , Zhang Jiahan , Yang Xiaofeng , Liu Tian , Kahn Shannon , Shu Hui‐Kuo , Tian Zhen
Published: February 2022

Purpose

Several inverse planning algorithms have been developed for Gamma Knife (GK) radiosurgery to determine a large number of plan parameters by solving an optimization problem, which typically consists of multiple objectives. The priorities among these objectives need to be repetitively adjusted to achieve a clinically good plan for each patient. This study aimed to achieve automatic and intelligent priority tuning by developing a deep reinforcement learning (DRL)‐based method to model the tuning behaviors of human planners.

Methods

We built a priority‐tuning policy network using deep convolutional neural networks. Its input was a vector composed of multiple plan metrics that were used in our institution for GK plan evaluation. The network can determine which tuning action to take based on the observed quality of the intermediate plan. We trained the network using an end‐to‐end DRL framework to approximate the optimal action‐value function. A scoring function was designed to measure the plan quality to calculate the received reward of a tuning action.

Results

Vestibular schwannoma was chosen as the test bed in this study. The number of training, validation and testing cases were 5, 5, and 16, respectively. For these three datasets, the average scores of the initial plans obtained with the same initial priority set were 3.63 ± 1.34, 3.83 ± 0.86 and 4.20 ± 0.78, respectively, while they were improved to 5.28 ± 0.23, 4.97 ± 0.44 and 5.22 ± 0.26 through manual priority tuning by human expert planners. Our network achieved competitive results with 5.42 ± 0.11, 5.10 ± 0. 42, 5.28 ± 0.20, respectively.

Conclusions

Our network can generate GK plans of comparable or slightly higher quality than the plans generated by human planners via manual priority tuning for vestibular schwannoma cases. The network can potentially be incorporated into the clinical workflow as planning assistance to improve GK planning efficiency and help to reduce plan quality variation caused by interplanner variability. We also hope that our method can reduce the workload of GK planners and allow them to spend more time on more challenging cases.
Cattaneo–Christov heat flux model for three‐dimensional magnetohydrodynamic flow of an Eyring Powell fluid over an exponentially stretching surface with convective boundary condition
Authors: Yusuf Tunde A. , Ashraf Muhaammad Bilal , Mabood Fazle
Published: February 2022

The present model concentrates on three‐dimensional steady incompressible flow of an Eyring‐Powell nanofluid past an exponentially stretching sheet with magnetic field. The Cattaneo–Christov heat flux with convective boundary condition is accounted for. Shooting method is the instrumental for obtaining numerical solution of the transformed‐converted system of the mathematical models. Behavior of the determining thermo‐physical parameters on the velocity, temperature, skin friction, heat transfer rate, and finally isotherms are considered. The major relevant outcomes of the current investigation are that increment in Eyring‐Powell parameter uplifts flow velocity, while that peters out the fluid temperature. Enhanced values of the mixed convection parameter weakened the skin friction coefficient while it slightly strengthened the rate of heat transfer.
Systematic Study on the Cytotoxic Potency of Commonly Used Dimeric Metal Precursors in Human Cancer Cell Lines
Authors: Geisler Heiko , Harringer Sophia , Wenisch Dominik , Urban Richard , Jakupec Michael A. , Kandioller Wolfgang , Keppler Bernhard K.
Published: February 2022

The cytotoxicities of seven dimeric metal species of the general formula [M(arene)Cl2]2, commonly used as precursors for complex synthesis and deemed biologically inactive, are investigated in seven commonly employed human cancer cell lines. Four of these complexes featured a ruthenium(II) core, where p‐cymene, toluene, benzene and indane were used as arenes. Furthermore, the osmium(II) p‐cymene dimer, as well as the Cp* dimers of rhodium(III) and its heavier analogue iridium(III) were included in this work (Cp*=1,2,3,4,5‐pentamethylcyclopentadienide). While the cytotoxic potencies of the ruthenium(II) and osmium(II) dimers are very low (or not even detectable at applicable concentrations), surprising activity, especially in cells from ovarian malignancies (with one or two‐digit micromolar IC50 values), have been found for the rhodium(III) and iridium(III) representatives. This publication is aimed at all researchers using synthetic procedures based on functionalization of these dimeric starting materials to rationalize changes in biological properties, especially cytotoxicity in cancer cells.
Hybrid nanocomposite packaging films from cellulose nanocrystals, zinc sulfide quantum dots reinforced polylactic acid with fluorescent and antibacterial properties
Authors: Shojaeiarani Jamileh , Shirzadifar Alimohammad , Shine Caleb , Reisi Amir Mohammad
Published: February 2022

In this study, nanocomposite thin films from cellulose nanocrystals (CNCs), zinc sulfide (ZnS) quantum dots, and polylactic acid (PLA) with fluorescent and antibacterial properties were constructed via a simple, and fast route. ZnS nanoparticles were added to CNC following the hydrothermal process. The film casting method was used to form thin nanocomposite films. The transmission electron microscope analysis revealed the uniform dispersion and ZnS nanoparticles through CNC suspension and FTIR results confirmed the interaction between –OH group on CNC and ZnS nanoparticles. A noticeable emission peak at 475 nm for CNC‐ZnS signifies the suitability of CNC‐ZnS for the assembly of ZnS QDs. The addition of ZnS‐coated CNC into PLA film resulted in a bright blue fluorescence when exposed to ultraviolet light. The incorporation of 5 wt% CNCs loaded with ZnS quantum dots into PLA notably decreased bacterial growth in Escherichia coli and Salmonella, respectively. Data obtained from dynamic mechanical analysis underlined that the presence of 1 wt% ZnS‐coated CNC in PLA increased the storage modulus of nanocomposite films by 25%. The CNCs loaded with ZnS quantum dots showed effective fluorescent and antimicrobial properties to be used as food packaging material.
Equitable practices in school mental health
Authors: DeBoer Jacob L. , Allouche Sam F. , Vasquez Jennifer I. , Rhodes Judith
Published: February 2022

Given the prevalence and negative outcomes associated with untreated mental health concerns, there is an urgent need for schools to enhance their capacity to prevent, identify, and address these needs. This is particularly important for students from culturally and linguistically diverse backgrounds, who are more likely to be underserved and face barriers to mental health treatment. Current methods of addressing mental health at school are reactive and insufficient, failing to address the needs of all students. Without an explicit focus on dismantling systemic racism and inequity within schools, our society will continue to underserve those already disenfranchised. As such, this paper outlines the challenges as well as the importance of incorporating culture into a conceptualization of universal mental health in schools. We include state‐, district‐, and school‐level solutions in this conceptualization. Specifically, we propose sample solutions to the implementation of equity‐focused, comprehensive mental health practices within a multitiered system of support, focusing predominantly on universal (i.e., tier 1 level) practices.
Bronchoscopy in the management of children from developing countries undergoing congenital heart surgery
Authors: Schnapper Michael , Dalal Ilan , Mandelberg Avigdor , Raucher Sternfeld Alona , Sasson Lior , Armoni Domany Keren
Published: February 2022

Objectives

To study the clinical characteristics and impact of bronchoscopy in children from developing countries, referred for cardiac surgery, through the "Save a Child's Heart" (SACH) organization.

Methods

We performed a retrospective hospital‐chart review of SACH children (0–18 years old) referred between 2006 and 2021 who underwent fiberoptic bronchoscopy. We examined demographics, congenital‐heart‐disease (CHD) types, bronchoscopy's indications and findings, subsequent recommendations, number of ventilation, and intensive‐care‐unit days. The primary outcome was percent changes in management and diagnosis, following the bronchoscopy. We included a control group matched‐for‐age and CHD type, who did not undergo bronchoscopy.

Results

We performed 82 bronchoscopies in 68 children: 18 (26.5%) preoperatively; 46 (67.6%) postoperatively; and four (5.9%) both. The most prevalent CHDs were Tetralogy‐of‐Fallot (27.9%) and ventricular‐septal‐defect (19.1%). The main indications were persistent atelectasis (41%) and mechanical ventilation/weaning difficulties (27.9%). Bronchoscopic evaluations revealed at least one abnormality in 51/68 (75%) children. The most common findings were external airway compression (23.5%), bronchomalacia (19.1%), and mucus secretions (14.7%). Changes in management were made in 35 (51.4%) cases, with a major change made in 14/35 (40%) children. Compared to the control group, the children undergoing bronchoscopy were both ventilated longer (median 6 vs. 1.5 days, p < 0.0001) and stayed longer in the intensive care unit (median 1.5 vs. 18.5 days, p < 0.0001).

Conclusion

A bronchoscopy is an important tool in the diagnosis and management of the unique group of children from developing countries with CHD referred for cardiac surgery. The results of our study, reveal a more complicated clinical course in children requiring bronchoscopy compared to controls.
Ultra‐Thermostability of Spatially Confined and Fully Protected Perovskite Nanocrystals by In Situ Crystallization
Authors: Li Qinyi , Shen Dongyang , Luo Chengzhao , Zheng Zhishuai , Xia Wenlin , Ma Wenchen , Li Jie , Yang Yixing , Chen Song , Chen Yu
Published: February 2022

Although all‐inorganic perovskite materials present multiple fascinating optical properties, their poor stability undermines their potential application in the field of multi‐color display. Herein, spatially confined CsPbBr3 nanocrystals are in situ crystallized within uniform mesoporous SiO2 nanospheres (MSNs) to regulate their size distribution, passivate their surface defects, shield them from water/oxygen, and more importantly, enhance their thermotolerance. As a result, the remnant PL intensity of the prepared spatially confined perovskite (CsPbBr3) nanocrystals by in situ crystallization within uniform mesoporous SiO2 nanospheres (SCP@MSNs) powders can be maintained over 98% of its initial value even after being immersed in harsh conditions (0.1 m HCl or 0.1 m NaOH) for 60 days. Furthermore, the prepared SCP@MSNs‐PDMS film demonstrates astonishing thermostability by maintaining almost consistent room temperature PL intensities after continuous heating–cooling cycles between 200 and 25 °C, which would greatly improve its processability during potential industrial manufacturing. The fabricated LCD backlit based on SCP@MSNs covers 124% of NTSC standard and 95.6% of Rec. 2020 standard, indicating its great potential in practical display field.
Towards an AI‐driven business development framework: A multi‐case study
Authors: John Meenu Mary , Olsson Helena Holmström , Bosch Jan
Published: February 2022

Artificial intelligence (AI) and the use of machine learning (ML) and deep learning (DL) technologies are becoming increasingly popular in companies. These technologies enable companies to leverage big quantities of data to improve system performance and accelerate business development. However, despite the appeal of ML/DL, there is a lack of systematic and structured methods and processes to help data scientists and other company roles and functions to develop, deploy and evolve models. In this paper, based on multi‐case study research in six companies, we explore practices and challenges practitioners experience in developing ML/DL models as part of large software‐intensive embedded systems. Based on our empirical findings, we derive a conceptual framework in which we identify three high‐level activities that companies perform in parallel with the development, deployment and evolution of models. Within this framework, we outline activities, iterations and triggers that optimize model design as well as roles and company functions. In this way, we provide practitioners with a blueprint for effectively integrating ML/DL model development into the business to achieve better results than other (algorithmic) approaches. In addition, we show how this framework helps companies solve the challenges we have identified and discuss checkpoints for terminating the business case.
Protective effect of nimbolide against streptozotocin induced gestational diabetes mellitus in rats via alteration of inflammatory reaction, oxidative stress, and gut microbiota
Authors: Ma Yifei , Xu Shan , Meng Juan , Li Lu
Published: February 2022

Background

Gestational diabetes mellitus (GDM) is a significant pregnancy‐related condition, which showed effect on the development of fetal. Anti‐inflammatory and antioxidant therapy commonly used for the treatment of GDM. Nimbolide already confirmed their anti‐inflammatory and anti‐oxidant effect against various animal disease model. Our objective in this research is to investigate the protective effect of nimbolide against STZ induced GDM and elucidate the mechanism.

Methods

In this experimental study, pregnant female Wistar rats were used and STZ (40 mg/kg) was used to induce the GDM. Blood glucose level (BGL), body weight and plasma insulin were assessed at regular time (gestational day 0, 9, and 18). Water intake, food intake, fecal and urine output were also estimated. In the female rats, hemoglobin (Hb), glycalated hemoglobin (HbA1c), hepatic glycogen, fructosamine, adiponectin, leptin, lipid, antioxidant and inflammatory cytokines parameters were estimated. In the fetuses, the fetues weight, implementation loss, and fetal weight were estimated. At the completion of the protocol, biochemical parameters were calculated. Gut microbiota was estimated in end of the study.

Results

Nimbolide treatment significantly (p < .001) improved the fetuses level and suppressed the fetal weight and implantation loss. Nimbolide treatment significantly (p < .001) suppressed the BGL and enhanced the body weight, insulin level. Nimbolide treatment suppressed the water intake, food intake, urinary and fecal output. Nimbolide significantly (p < .001) suppressed the fructosamine, leptin and enhanced the adiponectin level. Nimbolide treatment significantly (p < .001) decreased the malonaldehyde (MDA) level and boosted the total antioxidant capacity (TAC), superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione S‐transferase (GST) and catalase (CAT); suppressed the level of TNF‐α, IL‐1β, IL‐6, and boosted the level of IL‐10. Furthermore, nimbolide treatment reversed the gut microbiota alteration induced via STZ in female rats. At the phylum level, the Firmicutes and Bacteroidetes relative abundance was altered via nimbolide treatment. The ratio of F/B boosted in GDM group and nimbolide treatment significantly (p < .001) suppressed. Nimbolide considerably suppressed the firmicutes and enhanced the Bacteroidetes, CAG‐352, Lacnospirace.

Conclusion

Based on the findings, we may conclude that nimbolide protects the pregnant rats from GDM via alteration of inflammation, oxidative stress, and gut microbiota.
In Vitro Models for the Evaluation of Antisense Oligonucleotides in Skin
Authors: Bremer Jeroen , van den Akker Peter C.
Published: February 2022

The genodermatosis dystrophic epidermolysis bullosa (DEB) is caused by mutations in the COL7A1 gene which encodes type VII collagen (C7). In the cutaneous basement membrane zone, C7 secures attachment of the epidermal basal keratinocyte to the papillary dermis by means of anchoring fibril formation. The complete absence of these anchoring fibrils leads to severe blistering of skin and mucosa upon the slightest friction and early mortality. To date, although preclinical advances toward therapy are promising, treatment for the disease is merely symptomatic. Therefore, research into novel therapeutics is warranted.

Antisense oligonucleotide (ASO)-mediated exon skipping is such a therapy. Clinical examination of naturally occurring exon skipping suggested that this mechanism could most likely benefit the most severely affected patients. The severe form of DEB is caused by biallelic null mutations. Exon skipping aims to bind an ASO to the mutated exon of the pre-mRNA in the cell nucleus. Thereby, the ASO inhibits the recognition of the mutated exon by the splicing machinery, and as a result, the mutated exon is spliced out from the mRNA with its surrounding introns, i.e., it is skipped. Here, we describe in vitro methods to evaluate ASO-mediated exon skipping in a preclinical setting.
In Vitro Delivery of PMOs in Myoblasts by Electroporation
Authors: Goossens Remko , Aartsma-Rus Annemieke
Published: February 2022

Antisense oligonucleotides (AONs) are small synthetic molecules of therapeutic interest for a variety of human disease. Their ability to bind mRNA and affect its splicing gives AONs potential use for exon skipping therapies aimed at restoring the dystrophin transcript reading frame for Duchenne muscular dystrophy (DMD) patients. The neutrally charged phosphorodiamidate morpholino oligomers (PMOs) are a stable and relatively nontoxic AON modification. To assess exon skipping efficiency in vitro, it is important to deliver them to target cells. Here, we describe a method for the delivery of PMOs to myoblasts by electroporation. The described protocol for the Amaxa 4D X unit nucleofector system allows efficient processing of 16 samples in one nucleocuvette strip, aiding in high-throughput PMO efficacy screens.
Rapid Determination of MBNL1 Protein Levels by Quantitative Dot Blot for the Evaluation of Antisense Oligonucleotides in Myotonic Dystrophy Myoblasts
Authors: Moreno Nerea , González-Martínez Irene , Artero Rubén , Cerro-Herreros Estefanía
Published: February 2022

Western blot assays are not adequate for high-throughput screening of protein expression because it is an expensive and time-consuming technique. Here we demonstrate that quantitative dot blots in plate format are a better option to determine the absolute contents of a given protein in less than 48 h. The method was optimized for the detection of the Muscleblind-like 1 protein in patient-derived myoblasts treated with a collection of more than 100 experimental oligonucleotides.
Evaluation of Exon Skipping and Dystrophin Restoration in In Vitro Models of Duchenne Muscular Dystrophy
Authors: López-Martínez Andrea , Soblechero-Martín Patricia , Arechavala-Gomeza Virginia
Published: February 2022

Several exon skipping antisense oligonucleotides (eteplirsen, golodirsen, viltolarsen, and casimersen) have been approved for the treatment of Duchenne muscular dystrophy, but many more are in development targeting an array of different DMD exons. Preclinical screening of the new oligonucleotide sequences is routinely performed using patient-derived cell cultures, and evaluation of their efficacy may be performed at RNA and/or protein level. While several methods to assess exon skipping and dystrophin expression in cell culture have been developed, the choice of methodology often depends on the availability of specific research equipment.

In this chapter, we describe and indicate the relevant bibliography of all the methods that may be used in this evaluation and describe in detail the protocols routinely followed at our institution, one to evaluate the efficacy of skipping at RNA level (nested PCR) and the other the restoration of protein expression (myoblot), which provide good results using equipment largely available to most research laboratories.
Generation of Human iPSC-Derived Myotubes to Investigate RNA-Based Therapies In Vitro
Authors: Herrero-Hernandez Pablo , Bergsma Atze J. , Pijnappel W. W. M. Pim
Published: February 2022

Alternative pre-mRNAsplicing can be cell-type specific and results in the generation of different protein isoforms from a single gene. Deregulation of canonical pre-mRNAsplicing by disease-associated variants can result in genetic disorders. Antisense oligonucleotides (AONs) offer an attractive solution to modulate endogenous gene expression through alteration of pre-mRNAsplicing events. Relevant in vitro models are crucial for appropriate evaluation of splicing modifying drugs. In this chapter, we describe how to investigate the splicing modulating activity of AONs in an in vitro skeletal muscle model, applied to Pompe disease. We also provide a detailed description of methods to visualize and analyze gene expression in differentiated skeletal muscle cells for the analysis of muscledifferentiation and splicing outcome. The methodology described here is relevant to develop treatment options using AONs for other genetic muscle diseases as well, including Duchenne muscular dystrophy, myotonic dystrophy, and facioscapulohumeral muscular dystrophy.
Eye on a Dish Models to Evaluate Splicing Modulation
Authors: Hau Kwan-Leong , Lane Amelia , Guarascio Rosellina , Cheetham Michael E.
Published: February 2022

Inherited retinal dystrophies, such as Leber congenital amaurosis, Stargardt disease, and retinitis pigmentosa, are characterized by photoreceptor dysfunction and death and currently have few treatment options. Recent technological advances in induced pluripotent stem cell (iPSC) technology and differentiation methods mean that human photoreceptors can now be studied in vitro. For example, retinal organoids provide a platform to study the development of the human retina and mechanisms of diseases in the dish, as well as being a potential source for cell transplantation. Here, we describe differentiation protocols for 3D cultures that produce retinal organoids containing photoreceptors with rudimentary outer segments. These protocols can be used as a model to understand retinal disease mechanisms and test potential therapies, including antisense oligonucleotides (AONs) to alter gene expression or RNA processing. This “retina in a dish” model is well suited for use with AONs, as the organoids recapitulate patient mutations in the correct genomic and cellular context, to test potential efficacy and examine off-target effects on the translational path to the clinic.
Antisense RNA Therapeutics: A Brief Overview
Authors: Arechavala-Gomeza Virginia , Garanto Alejandro
Published: February 2022

Nucleic acid therapeutics is a growing field aiming to treat human conditions that has gained special attention due to the successful development of mRNA vaccines against SARS-CoV-2. Another type of nucleic acid therapeutics is antisense oligonucleotides, versatile tools that can be used in multiple ways to target pre-mRNA and mRNA. While some years ago these molecules were just considered a useful research tool and a curiosity in the clinical market, this has rapidly changed. These molecules are promising strategies for personalized treatments for rare genetic diseases and they are in development for very common disorders too. In this chapter, we provide a brief description of the different mechanisms of action of these RNA therapeutic molecules, with clear examples at preclinical and clinical stages.
Delivery of Antisense Oligonucleotides to the Mouse Retina
Authors: Garanto Alejandro
Published: February 2022

The eye is the organ in charge of vision and, given its properties, has become an excellent organ to test genetic therapies, including antisense oligonucleotide (AON) technology. In fact, the first AON receiving FDA and EMA approval was meant to treat an eye condition. Currently, dozens of clinical trials are being conducted for a variety of subtypes of inherited retinal disease. Although most of them are based on gene augmentation therapies, a phase 3 and two phase 1/2 clinical trials using AONs are ongoing. Since the retina is a layered structure of nondividing cells, obtaining human retinal tissue and expanding it in the lab is not possible, unless induced pluripotent stem cell technology is used. Mouse models have helped to elucidate the function of many genes, and the retinal structure is quite similar to that of humans. Thus, drug delivery to the mouse eye can provide valuable information for further optimization of therapies. In this chapter, the protocol for intravitreal injections of AONs is described in detail.
Preclinical Evaluation of the Renal Toxicity of Oligonucleotide Therapeutics in Mice
Authors: Echevarría Lucía , Goyenvalle Aurelie
Published: February 2022

Antisense oligonucleotides (ASO) therapeutics hold great promise for the treatment of numerous diseases, and several ASO drugs have now reached market approval, confirming the potential of this approach. However, some candidates have also failed, due to limited biodistribution/uptake and poor safety profile. In pursuit of better delivery and higher cellular uptake, ASO are being optimized, and new chemistries are developed or conjugated with various ligands. While these developments may lead to candidates with higher potency, it is important to keep the safety aspects in sight and screen for potential toxicity in early phases of preclinical development to avoid subsequent failure in clinical development. Our understanding of ASO-mediated toxicity keeps improving with increased preclinical and clinical data available. In this chapter, we will focus on the assessment of renal toxicity in mice and describe methods to measure the levels of general urinary biomarkers as well as acute kidney injury biomarkers following ASO treatment.
Determination of Optimum Ratio of Cationic Polymers and Small Interfering RNA with Agarose Gel Retardation Assay
Authors: Aydin Omer , Kanarya Dilek , Yilmaz Ummugulsum , Tunç Cansu Ümran
Published: February 2022

Nanomaterials have aroused attention in the recent years for their high potential for gene delivery applications. Most of the nanoformulations used in gene delivery are positively charged to carry negatively charged oligonucleotides. However, excessive positively charged carriers are cytotoxic. Therefore, the complexed oligonucleotide/nanoparticles should be well-examined before the application. In that manner, agarose gel electrophoresis, which is a basic method utilized for separation, identification, and purification of nucleic acid molecules because of its poriferous nature, is one of the strategies to determine the most efficient complexation rate. When the electric field is applied, RNA fragments can migrate through anode due to the negatively charged phosphate backbone. Because RNA has a uniform mass/charge ratio, RNA molecules run in agarose gel proportional according to their size and molecular weight. In this chapter, the determination of complexation efficiency between cationic polymer carriers and small interfering RNA (siRNA) cargos by using agarose gel electrophoresis is described. siRNA/cationic polymer carrier complexes are placed in an electric field and the charged molecules move through the counter-charged electrodes due to the phenomenon of electrostatic attraction. Nucleic acid cargos are loaded to cationic carriers via the electrostatic interaction between positively charged amine groups (N) of the carrier and negatively charged phosphate groups (P) of RNA. The N/P ratio determines the loading efficiency of the cationic polymer carrier. In here, the determination of N/P ratio, where the most efficient complexation occurs, by exposure to the electric field with a gel retardation assay is explained.
Development and Use of Cellular Systems to Assess and Correct Splicing Defects
Authors: Suárez-Herrera Nuria , Tomkiewicz Tomasz Z. , Garanto Alejandro , Collin Rob W. J.
Published: February 2022

A significant proportion of mutations underlying genetic disorders affect pre-mRNA splicing, generally causing partial or total skipping of exons, and/or inclusion of pseudoexons. These changes often lead to the formation of aberrant transcripts that can induce nonsense-mediated decay, and a subsequent lack of functional protein. For some genetic disorders, including inherited retinal diseases (IRDs), reproducing splicing dynamics in vitro is a challenge due to the specific environment provided by, e.g. the retinal tissue, cells of which cannot be easily obtained and/or cultured. Here, we describe how to engineer splicing vectors, validate the reliability and reproducibility of alternative cellular systems, assess pre-mRNA splicing defects involved in IRD, and finally correct those by using antisense oligonucleotide-based strategies.
Introduction: The Pandemic of Argumentation
Authors: Oswald Steve , Lewiński Marcin , Greco Sara , Villata Serena
Published: February 2022

This chapter introduces the volume and contextualises its scope, which covers communicative aspects of the current COVID-19 pandemic as well as the epidemic of misinformation from the perspective of argumentation theory. Argumentation theory is uniquely placed to understand and account for the challenges of public reason as expressed through argumentative discourse. The book thus focuses on the extent to which the forms, norms and functions of public argumentation have changed in the face of the COVID-19 pandemic. This question is investigated along descriptive, normative and prescriptive research lines at the core of the COST Action project CA 17132: European network for Argumentation and Public PoLicY analysis (APPLY). Contributions are divided into three groups, which (i) examine various features and aspects of public and institutional discourse about the COVID-19 pandemic, (ii) scrutinise the way health policies have been discussed, debated, attacked and defended in the public sphere, and (iii) consider a range of measures meant to improve the quality of public discourse, and public deliberation in particular, in such a way that concrete proposals for argumentative literacy are brought to light.
The UK Government’s “Balancing Act” in the Pandemic: Rational Decision-Making from an Argumentative Perspective
Authors: Fairclough Isabela
Published: February 2022

This chapter looks at how the “balance” between lives, livelihoods and other concerns was talked about in four main newspapers in the UK, between March 2020 and March 2021, in assessing the UK government’s performance. Different arguments were made for opposite conclusions, favouring either strict and prolonged lockdowns or, on the contrary, a speedy exit from lockdown and a resumption of normal life. From the point of view of argumentation theory, the empirical data suggests that what is being balanced or weighed together in pro/con argumentation by two opposite parties are not as much the costs and benefits of one’s own proposal, but the costs of one proposal against the costs of its alternative (a “cost-cost” analysis). Rather than defending their own proposal by arguing that the benefits outweigh the costs, each side is criticizing the opponent’s proposal by claiming that the costs of their proposal are more unacceptable than the costs of their own. An implicit minimax strategy (minimize costs in a worst-case scenario) was applied in different ways, depending on how the consequences were assessed, and how this assessment changed over time. The debate over lockdown illustrated an interesting type of pro/con argument, typical to crisis situations, in which all the intended “benefits” were in fact avoided “costs”, and contrasted a medical/epidemiological perspective with a political perspective on the best course of action.
Justification of Decision-Making in Response to COVID-19 Socio-Scientific Dilemmas
Authors: Dalyot Keren , Rozenblum Yael , Baram-Tsabari Ayelet
Published: February 2022

Argumentation skills are important for informed decision-making, especially in everyday life when engaging with science. The onset of the COVID-19 pandemic is an ideal opportunity to study laypeople’s use of argumentation skills when engaging with a scientific issue daily, while making relevant decisions that affect their families and society. This study frames the pandemic as a Socio-Scientific Issue (SSI)—a scientific issue with links to several social science disciplines (economics, politics, and sociology). The current study explores decision making and argumentation in the context of COVID-19 among the Israeli public as well as the connection between demographic characteristics, scientific knowledge and education and the quality of their argumentations. An online survey to examine responses to 2 specifically designed social dilemmas was conducted in April 2020 (n = 439). Our findings suggest that laypeople tend to use justifications that were classified as ‘scientific argumentation’ but we could not demonstrate a connection between demographic characteristics, scientific knowledge and decision making. We did find a positive connection between peoples’ perception of control over the situation and their compliance with the official guidelines. As a relevant Socio-Scientific Issue (SSI), COVID-19 stretched to the limit the need for public argumentation with changing scientific and medical information.