bluuebunny/crossref_metadata_2025 huggingface dataset:
RNA Extraction from Plant Tissue with Homemade Acid Guanidinium Thiocyanate Phenol Chloroform (AGPC)
Authors: Zepeda Baltasar , Verdonk Julian C.
Published: January 2022

Gene expression studies are a powerful technique to study biological processes, and isolating RNA that is pure, intact, and in sufficient amounts for downstream applications is key. Over the years, the field has moved to the use of commercial kits and ready‐made extraction buffers for RNA isolation. This became particularly problematic during the COVID‐19 crisis when supply chains were affected and when RNA extraction and analysis reagents were suddenly scarce at a time when they were particularly required. Acid guanidinium thiocyanate‐phenol‐chloroform (AGPC) is one of the oldest RNA extraction solutions, in use since 1987. It is known as a ready‐made solution, sold under different brand names, and is typically the most expensive reagent in the RNA extraction process. In this article, we describe how to prepare a low‐cost homemade AGPC solution and provide tips on how to use it for obtaining high‐quality RNA, as well as describe possible modifications for different conditions. The protocol is based on a phase separation, where RNA is maintained in the aqueous phase and DNA and proteins remain in the interphase and organic phase. After cleaning, precipitation, and resuspension steps, the RNA is ready to be quantified and used for downstream applications. By following this protocol, good yields of high‐quality RNA can be obtained from a wide variety of tissues and organisms, and we exemplify the approach here using plant tissues. Some plant tissues contain extra interferents (such as sugars), and for high‐quality RNA isolation from those tissues, an alternate protocol is provided. © 2022 The Authors. Current Protocols published by Wiley Periodicals LLC.

Basic Protocol: RNA isolation with homemade acid guanidinium thiocyanate‐phenol‐chloroform (AGPC)

Alternate Protocol: RNA isolation from high carbohydrate‒containing tissues using an NTES‐AGPC combination
Chromatin Immunoprecipitation in Chloroplasts
Authors: Wang Wenjie , Cheng Lingling , Sun Qianwen
Published: January 2022

Chromatin is the genetic material assembled by nucleic acids (including DNA and RNA) and proteins. The biological functions of chromatin are highly dependent on the interaction between DNA (and/or RNA) and proteins that bind to it. Chromatin immunoprecipitation (ChIP) is a powerful technique for evaluating these interactions and has been widely used to characterize the functions of nuclear proteins. However, its application in identifying plant organellar chromatin‐binding proteins is lagging. This article describes the method for analyzing the association of chloroplast‐localized proteins with the chloroplast genome. © 2022 Wiley Periodicals LLC.

Basic Protocol 1: Chloroplast isolation

Basic Protocol 2: Crosslinking of DNA–Protein complexes

Basic Protocol 3: Chromatin isolation and preparation

Support Protocol: Bead‐antibody complex preparation

Basic Protocol 4: Immunoprecipitation and washes

Basic Protocol 5: DNA preparation

Basic Protocol 6: Analysis of results
EU financial regulations and social impact measurement practices: A comprehensive framework on finance for sustainable development
Authors: Bengo Irene , Boni Leonardo , Sancino Alessandro
Published: January 2022

Sustainability is becoming the main character of the financial industry in Europe, especially after the Sustainable Finance Disclosure Regulation (SFDR) 2019/2088, which came into force on March 10th, 2021. However, despite the top‐down indications for disclosing and reporting sustainability practices provided by this new policy, financial actors still lack a comprehensive framework on how to track and measure their social and environmental contributions within the perimeter of this novel institutional context. This paper discusses the implications for financial actors brought by the SDFR and builds a conceptual link with social impact measurement practices. In particular, the article provides a comprehensive framework that identifies strategic approaches and measurement tools for financial actors for building a more sustainable finance, that is a finance focused on the purest dimension of blended value and more attentive to sustainable development.
Synthesizing Mechanism of the Mn4Ca Cluster Mimicking the Oxygen‐Evolving Center in Photosynthesis
Authors: Chen Changhui , Xu Boran , Yao Ruoqing , Chen Yang , Zhang Chunxi
Published: January 2022

The photosynthetic oxygen‐evolving center (OEC) is a unique Mn4CaO5 cluster that serves as a blueprint to develop superior water‐splitting catalysts for the generation of solar fuels in artificial photosynthesis. It is a great challenge and long‐standing issue to reveal the synthesizing mechanism of this Mn4CaO5 cluster in both natural and artificial photosynthesis. Herein, efforts were made to reveal the synthesizing mechanism of an artificial Mn4CaO4 cluster, a close mimic of the OEC. Four key intermediates were successfully isolated and structurally characterized for the first time. It was demonstrated that the Mn4CaO4 cluster could be formed through a reaction between a thermodynamically stable Mn3CaO4 cluster and an unusual four‐coordinated MnIII ion, followed by stabilization process through binding an organic base (e.g., pyridine) on the “dangling” Mn ion. These findings shed new light on the synthesizing mechanism of the OEC and rational design of new artificial water‐splitting catalysts.
Novel hair snare and genetic methods for non‐invasive bobcat detection
Authors: Rounsville Thomas F. , Rogers Richard E. , Welsh Amy B. , Ryan Christopher W. , Anderson James T.
Published: January 2022

Over the past 20 years, the use of non‐invasive hair snare surveys in wildlife research and management has become more prevalent. While these tools have been used to answer important research questions, these techniques often fail to gather information on elusive carnivores, such as bobcats (Lynx rufus). Due to the limited success of previous bobcat studies using hair snares which required active rubbing, this technique has largely fallen out of use, in favor of camera trapping. The goal of our study was to construct a novel, passive bobcat hair snare that could be deployed regardless of terrain or vegetation features, which would be effective for use in capture–recapture population estimation at a large spatial scale. This new hair snare was deployed in 1500 10‐km2 cells across West Virginia (USA) between two sampling seasons (2015–2016). Collected hair samples were analyzed with newly developed mitochondrial DNA primers specifically for felids and qPCR to determine species of origin, with enough sensitivity to identify samples as small as two bobcat hairs. Over the two years of the study, a total of 378 bobcat detections were recorded from 42,000 trap nights of sampling, for an overall rate of 0.9 detections/100 trap nights—nearly 2–6 times greater than any previous bobcat hair snare study. While the overall number of recaptured animals was low (n = 9), continued development of this platform should increase its usefulness in capture–recapture studies.
Effects of climate change and pollen supplementation on the reproductive success of two grassland plant species
Authors: Andrzejak Martin , Korell Lotte , Auge Harald , Knight Tiffany M.
Published: January 2022

Climate change has the potential to alter plant reproductive success directly and indirectly through disruptions in animal pollination. Climate models project altered seasonal precipitation patterns, and thus, the effects of climate change on available resources and pollination services will depend on the season. Plants have evolved reproductive strategies to so they are not limited by either pollen or water availability in their reproductive success, and therefore, we expect that the disruption of climate change might cause plants to be more pollen limited in seasons that become wetter than they were historically. In this study, we conducted a pollen supplementation experiment within the Global Change Experiment Facility (GCEF) in Central Germany. The GCEF experimentally manipulates future climate based on a realistic scenario of climate change for the region (drier summers and wetter springs and falls) in a native grassland ecosystem. We quantified seed production of two perennial species Dianthus carthusianorum and Scabiosa ochroleuca in response to pollination treatments (control, supplement), climate treatments (ambient and future) and season (summer and fall). Dianthus carthusianorum produced more seeds in future climate conditions independent of the season, but only when given supplemental pollen. Both species showed an increased reproduction in summer compared with the fall. We did not find evidence for our specific expectation of higher pollen limitation in the future climate and fall season (i.e., no three‐way interaction pollination × season × climate), which might be explained by the high‐drought tolerance and generalized pollination of our focal plant species. We conclude that plant reproductive success has the potential to change with changing climates and that this change will depend on how pollinator services change in the future. We offer many suggestions for future studies that are necessary to understand the context dependence and underlying mechanisms of plant reproductive responses to climate.
Comparing genome scans among species of the stickleback order reveals three different patterns of genetic diversity
Authors: Reeve James , Li Qiushi , Lindtke Dorothea , Yeaman Samuel
Published: January 2022

Comparing genome scans among species is a powerful approach for investigating the patterns left by evolutionary processes. In particular, this offers a way to detect candidate genes that drive convergent evolution. We compared genome scan results to investigate if patterns of genetic diversity and divergence are shared among divergent species within the stickleback order (Gasterosteiformes): the threespine stickleback (Gasterosteus aculeatus), ninespine stickleback (Pungitius pungitus), and tubesnout (Aulorhynchus flavidus). Populations were sampled from the southern and northern edges of each species’ range, to identify patterns associated with latitudinal changes in genetic diversity. Weak correlations in genetic diversity (FSTand expected heterozygosity) and three different patterns in the genomic landscape were found among these species. Additionally, no candidate genes for convergent evolution were detected. This is a counterexample to the growing number of studies that have shown overlapping genetic patterns, demonstrating that genome scan comparisons can be noisy due to the effects of several interacting evolutionary forces.
Intergrading reef communities across discrete seaweed habitats in a temperate–tropical transition zone: Lessons for species reshuffling in a warming ocean
Authors: Mulders Yannick , Filbee‐Dexter Karen , Bell Sahira , Bosch Nestor E. , Pessarrodona Albert , Sahin Defne , Vranken Sofie , Zarco‐Perello Salvador , Wernberg Thomas
Published: January 2022

Temperate reefs are increasingly affected by the direct and indirect effects of climate change. At many of their warm range edges, cool‐water kelps are decreasing, while seaweeds with warm‐water affinities are increasing. These habitat‐forming species provide different ecological functions, and shifts to warm‐affinity seaweeds are expected to modify the structure of associated communities. Predicting the nature of such shifts at the ecosystem level is, however, challenging, as they often occur gradually over large geographical areas. Here, we take advantage of a climatic transition zone, where cool‐affinity (kelp) and warm‐affinity (Sargassum) seaweed forests occur adjacently under similar environmental conditions, to test whether these seaweed habitats support different associated seaweed, invertebrate, coral, and fish assemblages. We found clear differences in associated seaweed assemblages between habitats characterized by kelp and Sargassum abundance, with kelp having higher biomass and seaweed diversity and more cool‐affinity species than Sargassum habitats. The multivariate invertebrate and fish assemblages were not different between habitats, despite a higher diversity of fish species in the Sargassum habitat. No pattern in temperature affinity of the invertebrate or fish assemblages in each habitat was found, and few fish species were exclusive to one habitat or the other. These findings suggest that, as ocean warming continues to replace kelps with Sargassum, the abundance and diversity of associated seaweeds could decrease, whereas fish could increase. Nevertheless, the more tropicalized seaweed habitats may provide a degree of functional redundancy to associated fauna in temperate seaweed habitats.
Tumor hijacks macrophages and microbiota through extracellular vesicles
Authors: Jiang Jipeng , Mei Jie , Ma Yongfu , Jiang Shasha , Zhang Jian , Yi Shaoqiong , Feng Changjiang , Liu Yang , Liu Ying
Published: January 2022

The tumor microenvironment (TME) is a biological system with sophisticated constituents. In addition to tumor cells, tumor‐associated macrophages (TAMs) and microbiota are also dominant components. The phenotypic and functional changes of TAMs are widely considered to be related to most tumor progressions. The chronic colonization of pathogenic microbes and opportunistic pathogens accounts for the generation and development of tumors. As messengers of cell‐to‐cell communication, tumor‐derived extracellular vesicles (TDEVs) can transfer various malignant factors, regulating physiological and pathological changes in the recipients and affecting TAMs and microbes in the TME. Despite the new insights into tumorigenesis and progress brought by the above factors, the crosstalk among tumor cells, macrophages, and microbiota remain elusive, and few studies have focused on how TDEVs act as an intermediary. We reviewed how tumor cells recruit and domesticate macrophages and microbes through extracellular vehicles and how hijacked macrophages and microbiota interact with tumor‐promoting feedback, achieving a reciprocal coexistence under the TME and working together to facilitate tumor progression. It is significant to seek evidence to clarify those specific interactions and reveal therapeutic targets to curb tumor progression and improve prognosis.
Engineered cancer cell membranes: An emerging agent for efficient cancer theranostics
Authors: Guo Yunqi , Wang Zhiqiang , Shi Xiangyang , Shen Mingwu
Published: January 2022

For efficient cancer theranostics, surface modification of nanomaterials plays an important role in improving targeting ability and reducing the non‐specific interactions with normal tissues. Recently, the biomimetic technology represented by coating of cancer cell membranes (CCMs) has been regarded as a promising method to strengthen the biocompatibility and targeting specificity of nanomaterials. Furthermore, the engineered CCMs (ECCMs) integrated with the natural biological properties of CCMs and specific functions from other cells or proteins have offered more possibilities in the field of cancer theranostics. Herein, the recent progresses in the design and preparation of ECCMs are summarized, and the applications of ECCMs in targeting delivery, activation of immunity, and detection of circulating tumor cells are reviewed. Finally, the current challenges and future perspectives with regard to the development of ECCMs are briefly discussed.
Onion (Allium cepa L.) bioactives: Chemistry, pharmacotherapeutic functions, and industrial applications
Authors: Sagar Narashans Alok , Pareek Sunil , Benkeblia Noureddine , Xiao Jianbo
Published: January 2022

Bioactive compounds possess different health benefits. Onion contains various bioactive compounds, such as organosulfur compounds, flavonols, ascorbic acids, and carbohydrate prebiotics, and its by‐products have more content of flavonoids than the bulb. Diallyl monosulfide, diallyl disulfide, diallyl trisulfide, and diallyl tetrasulfide are the major organosulfur compounds, whereas quercetin, kaempferol, anthocyanin, and luteolin are considered as main flavonoids. Ascorbic acid and fructooligosaccharides are also regarded as bioactive compounds. Onion bioactive compounds have the strong antioxidant potential for neutralizing oxidative stress of the cells. These bioactive components are beneficial as anticarcinogenic, antibiotic, anti‐inflammatory, antiplatelet, antidiabetic, and cardioprotective agents along with other nutritional benefits. However, various postharvest practices have an impact on these bioactive compounds, for example, curing mostly enhances the bioactive level and processing temperature generally decreases the concentration of many of them, whereas storage studies suggest an increase of others under optimized conditions. Additionally, conventional extraction techniques showed a negative impact on bioactive compounds of onion, whereas innovative methods yielded a higher amount of bioactive components. There is a need for innovative and integrated procedures in the postharvest sector to maintain or enhance the level of bioactive compounds without compromising the quality of onions. The present review comprehensively describes different bioactive compounds of onion, their chemistry, and their pharmacotherapeutic roles. Moreover, it also explores the effects of various postharvest factors, such as temperature, storage duration, and extraction conditions on the level of the bioactive components. It also suggests industrial applications of onion waste and its bioactive compounds in the food sector.
mGluR5 and GABAA receptor‐specific parametric PET atlas construction—PET/MR data processing pipeline, validation, and application
Authors: Kaulen Nicolas , Rajkumar Ravichandran , Régio Brambilla Cláudia , Mauler Jörg , Ramkiran Shukti , Orth Linda , Sbaihat Hasan , Lang Markus , Wyss Christine , Rota Kops Elena , Scheins Jürgen , Neumaier Bernd , Ermert Johannes , Herzog Hans , Langen Karl‐Joseph , Lerche Christoph , Shah N. Jon , Veselinović Tanja , Neuner Irene
Published: January 2022

The glutamate and γ‐aminobutyric acid neuroreceptor subtypes mGluR5 and GABAA are hypothesized to be involved in the development of a variety of psychiatric diseases. However, detailed information relating to their in vivo distribution is generally unavailable. Maps of such distributions could potentially aid clinical studies by providing a reference for the normal distribution of neuroreceptors and may also be useful as covariates in advanced functional magnetic resonance imaging (MR) studies. In this study, we propose a comprehensive processing pipeline for the construction of standard space, in vivo distributions of non‐displaceable binding potential (BPND), and total distribution volume (VT) based on simultaneously acquired bolus‐infusion positron emission tomography (PET) and MR data. The pipeline was applied to [11C]ABP688‐PET/MR (13 healthy male non‐smokers, 26.6 ± 7.0 years) and [11C]Flumazenil‐PET/MR (10 healthy males, 25.8 ± 3.0 years) data. Activity concentration templates, as well as VT and BPND atlases of mGluR5 and GABAA, were generated from these data. The maps were validated by assessing the percent error δ from warped space to native space in a selection of brain regions. We verified that the average δABP = 3.0 ± 1.0% and δFMZ = 3.8 ± 1.4% were lower than the expected variabilities σ of the tracers (σABP = 4.0%–16.0%, σFMZ = 3.9%–9.5%). An evaluation of PET‐to‐PET registrations based on the new maps showed higher registration accuracy compared to registrations based on the commonly used [15O]H2O‐template distributed with SPM12. Thus, we conclude that the resulting maps can be used for further research and the proposed pipeline is a viable tool for the construction of standardized PET data distributions.
The differences in distant metastatic patterns and their corresponding survival between thyroid cancer subtypes
Authors: Vuong Huy Gia , Le Minh‐Khang , Hassell Lewis , Kondo Tetsuo , Kakudo Kennichi
Published: January 2022

Introduction

This study aimed to systematically elucidate the metastatic patterns and their corresponding survival of each thyroid cancer subtype at time of diagnosis.

Methods

We accessed the Surveillance, Epidemiology, and End Results (SEER) database from 2010 to 2018 to search for primary thyroid cancers with DM at presentation (M1).

Results

We included 2787 M1 thyroid cancers for statistical analyses and the incidence of DM at presentation was 2.4%. Lung was the most common metastatic site for anaplastic thyroid carcinoma (ATC), poorly differentiated thyroid carcinoma (PDTC), papillary thyroid carcinoma (PTC), and oncocytic (Hurthle) cell carcinoma (HCC) whereas bone is the favorable disseminated site of follicular thyroid carcinoma (FTC) and medullary thyroid carcinoma (MTC). Patients with multi‐organ metastases had the worst survival whereas bone metastases were associated with a favorable outcome (p < 0.001).

Conclusion

There are significant differences in DM patterns of thyroid cancer subtypes and their corresponding survival.
Therapeutic efficacy of FASN inhibition in preclinical models of HCC
Authors: Wang Haichuan , Zhou Yi , Xu Hongwei , Wang Xue , Zhang Yi , Shang Runze , O'Farrell Marie , Roessler Stephanie , Sticht Carsten , Stahl Andreas , Evert Matthias , Calvisi Diego F. , Zeng Yong , Chen Xin
Published: January 2022

Background and Aims

Aberrant activation of fatty acid synthase (FASN) is a major metabolic event during the development of HCC. We evaluated the therapeutic efficacy of TVB3664, a FASN inhibitor, either alone or in combination, for HCC treatment.

Approach and Results

The therapeutic efficacy and the molecular pathways targeted by TVB3664, either alone or with tyrosine kinase inhibitors or the checkpoint inhibitor anti–programmed death ligand 1 antibody, were assessed in human HCC cell lines and multiple oncogene‐driven HCC mouse models. RNA sequencing was performed to elucidate the effects of TVB3664 on global gene expression and tumor metabolism. TVB3664 significantly ameliorated the fatty liver phenotype in the aged mice and AKT‐induced hepatic steatosis. TVB3664 monotherapy showed moderate efficacy in NASH‐related murine HCCs, induced by loss of phosphatase and tensin homolog and MET proto‐oncogene, receptor tyrosine kinase (c‐MET) overexpression. TVB3664, in combination with cabozantinib, triggered tumor regression in this murine model but did not improve the responsiveness to immunotherapy. Global gene expression revealed that TVB3664 predominantly modulated metabolic processes, whereas TVB3664 synergized with cabozantinib to down‐regulate multiple cancer‐related pathways, especially the AKT/mammalian target of rapamycin pathway and cell proliferation genes. TVB3664 also improved the therapeutic efficacy of sorafenib and cabozantinib in the FASN‐dependent c‐MYC‐driven HCC model. However, TVB3664 had no efficacy nor synergistic effects in FASN‐independent murine HCC models.

Conclusions

This preclinical study suggests the limited efficacy of targeting FASN as monotherapy for HCC treatment. However, FASN inhibitors could be combined with other drugs for improved effectiveness. These combination therapies could be developed based on the driver oncogenes, supporting precision medicine approaches for HCC treatment.
The DESC catchments: Long‐term monitoring of inland Precambrian shield catchment streamflow and water chemistry in Central Ontario, Canada
Authors: James April L. , Yao Huaxia , McConnell Christopher , Field Timothy , Yang Yinan
Published: January 2022

Since the early 1970s, the Dorset Environmental Science Centre (DESC) research catchments have been home to long‐term monitoring and study of terrestrial headwater catchment processes, their linkages to inland aquatic ecosystems and the influence of both natural variation and human activities. Located on the Precambrian Shield in central Ontario, Canada, covered by mixed Great Lakes‐St. Lawrence forest, the 29 catchments, defined by inflows and outflows to eight lakes, have been monitored for streamflow, meteorology and water chemistry, with long‐term datasets spanning from 1976 to the present. These datasets have provided insights into cold region hydrologic processes such as runoff generation, wetland and groundwater–surface water interactions, snow and ice processes, and catchment linkages to lake nutrient budgets and ecology. The datasets have supported catchment transit time estimates, hydrological modelling and cold region intercomparison studies. Starting with early research efforts driven by concerns over impacts from cottage development and acid deposition on soils, rivers and lakes, the DESC catchment datasets have supported study of impacts of stressors of forest harvesting, calcium depletion, road salt application and climate change. Ongoing monitoring of streamflow, meteorology and water chemistry in the DESC catchments continues to offer unique opportunities for investigation of critical zone processes in Precambrian shield catchments, their model representation and anthropogenic impacts.
Use of external cephalic version in Portuguese public hospitals
Authors: Pinto Luísa , Clode Nuno , Ayres‐de‐Campos Diogo
Published: January 2022

Objective

To evaluate the use of external cephalic version (ECV) in Portuguese public hospitals with maternity services, as well as exploring the main motives for not offering the technique.

Methods

A cross‐sectional observational study was conducted involving an online survey with 34 questions, accessed via an email addressed to all Heads of Department of Portuguese state‐owned hospitals with maternity services. In centers where the technique was performed, information was requested on success rates, contraindications for the procedure, and practical aspects related to its use. In centers where ECV was not offered, the underlying reasons for this were queried.

Results

Answers were received from 41 out of the 43 state hospitals with maternity services (95.3%). Sixteen hospitals perform the technique (39%), with reported annual numbers ranging from 3 to 51, and success rates ranging from 25% to 85% (12 respondents). The main reasons for not offering the technique were lack of experience and lack of conditions to perform it safely. Most centers (87.8%) reported that they would welcome hands‐on training in ECV.

Conclusion

ECV is used in a minority of Portuguese state‐owned hospitals. Efforts are needed to achieve a wider implementation of the technique, with a particular focus on simulation‐based training.
Rational design of anti‐inflammatory lipid nanoparticles for mRNA delivery
Authors: Zhang Hanwen , Han Xuexiang , Alameh Mohamad‐Gabriel , Shepherd Sarah J. , Padilla Marshall S. , Xue Lulu , Butowska Kamila , Weissman Drew , Mitchell Michael J.
Published: January 2022

Lipid nanoparticles (LNPs) play a crucial role in delivering messenger RNA (mRNA) therapeutics for clinical applications, including COVID‐19 mRNA vaccines. While mRNA can be chemically modified to become immune‐silent and increase protein expression, LNPs can still trigger innate immune responses and cause inflammation‐related adverse effects. Inflammation can in turn suppress mRNA translation and reduce the therapeutic effect. Dexamethasone (Dex) is a widely used anti‐inflammatory corticosteroid medication that is structurally similar to cholesterol, a key component of LNPs. Here, we developed LNP formulations with anti‐inflammatory properties by partially substituting cholesterol with Dex as a means to reduce inflammation. We demonstrated that Dex‐incorporated LNPs effectively abrogated the induction of tumor necrosis factor alpha (TNF‐ɑ) in vitro and significantly reduced its expression in vivo. Reduction of inflammation using this strategy improved in vivo mRNA expression in mice by 1.5‐fold. Thus, we envision that our Dex‐incorporated LNPs could potentially be used to broadly to reduce the inflammatory responses of LNPs and enhance protein expression of a range of mRNA therapeutics.
Molecularly aligned films of [1]benzothieno[3,2‐b][1]benzothiophene derivatives by solution shearing: Effect of alkyl substitution on morphology and charge transporting property
Authors: Prakoso Suhendro Purbo , Ke Yao‐Jin , Huang Ding‐Chi , Wang Chien‐Lung , Tao Yu‐Tai
Published: January 2022

The self‐assembly behaviors, crystal morphologies, and charge transporting properties of π–conjugated framework can be strongly affected by the functional group/substituent it carries, as well as the way the film is prepared. In this work, thin films of mono‐ and di‐alkylated [1]benzothieno[3,2‐b][1]‐benzothiophene (BTBT) derivatives were prepared on a silicon substrate by solution‐shearing method. Their film morphologies and crystal packing details were examined using polarized optical microscope, grazing incident wide‐angle X‐ray scattering, and atomic force microscope. With 2‐tridecyl[1]benzothieno[3,2‐b][1]‐benzothiophene (C13‐BTBT), a film with well‐aligned molecules packed in face‐to‐face π‐stacking arrangement was obtained, rendering a 1D‐like charge conduction channel. For 2,7‐bis‐tridecyl[1]benzothieno[3,2‐b][1]‐benzothiophene (C13‐BTBT‐C13), well‐aligned molecules packed in a layered‐herringbone arrangement were obtained. With these films as a conducting channel in transistor fabrication, high‐field‐effect mobility was realized for C13‐BTBT‐C13, with a maximum mobility (μh) of 0.645 cm2 /Vs and low anisotropic effect, presumably owing to the 2D growth of the π–conjugated skeleton in LHB packing. This study demonstrated the advantage of 2D semiconducting channels formed from C13‐BTBT‐C13 by low‐cost and scalable solution‐shearing processes, necessary for practical use in industry‐level and future flexible electronics.
Medication Optimization Using Pharmacogenomic Testing in a Complex Mental Health Population Prescribed Psychiatric Polypharmacy
Authors: Wood Amanda Ernst , Agrawal Deepika , Deem Alison P. , Dupper Knoper Terri L. , Merino Rosa F. , Molzof Hylton E. , Maus Laurie E. , Kim Floreen , Lodin Zohra , Lim Sonia
Published: January 2022

The use of polypharmacy has become significantly more common over the past two decades, increasing the risk of drug–drug interactions and adverse drug reactions. Pharmacogenomic (PGx) assays have the purported benefit of being able to predict an individual's response to a specific medication based on genetic markers, which may facilitate the development of optimized medication regimens for patients prescribed polypharmacy. This 12‐week pilot study examined the impact of the PGx results on the clinical management of Veterans who were prescribed psychiatric polypharmacy. Psychiatric medication providers were given access to the PGx assay results, including notification of drug–drug–gene interactions computed from an algorithm decision tool, to assist with medication management decisions. Veteran outpatients (N = 53) prescribed polypharmacy (mean = 13.15 medications) were enrolled into the study. In 92.4% of cases, providers changed medications at baseline, with 83% of providers indicating that they changed their original medication plan based on the PGx results. Clinical improvement over the 12‐week treatment phase was seen in depression (F(1.63, 45) = 5.45, P = .01, η2 = .11) and mental health quality of life (F(2.00, 45) = 4.16, P < .05, η2 = .16). Adverse drug effects were unchanged or improved over time. Rates of polypharmacy remained unchanged. The results suggest that medication changes based on the PGx assay may be beneficial in a complex patient population prescribed polypharmacy.
Impact of protocol‐based physiotherapy on insulin sensitivity and peripheral glucose metabolism in critically ill patients
Authors: Carbon Niklas M. , Engelhardt Lilian J. , Wollersheim Tobias , Grunow Julius J. , Spies Claudia D. , Märdian Sven , Mai Knut , Spranger Joachim , Weber‐Carstens Steffen
Published: January 2022

Background

The impact of physiotherapy on insulin sensitivity and peripheral glucose metabolism in critically ill patients is not well understood.

Methods

This pooled analysis investigates the impact of different physiotherapeutic strategies on insulin sensitivity in critically ill patients. We pooled data from two previous trials in adult patients with sequential organ failure assessment score (SOFA)≥ 9 within 72 h of intensive care unit (ICU) admission, who received hyperinsulinaemic euglycaemic (HE) clamps. Patients were divided into three groups: standard physiotherapy (sPT, n = 22), protocol‐based physiotherapy (pPT, n = 8), and pPT with added muscle activating measures (pPT+, n = 20). Insulin sensitivity index (ISI) was determined by HE clamp. Muscle metabolites lactate, pyruvate, and glycerol were measured in the M. vastus lateralis via microdialysis during the HE clamp. Histochemical visualization of glucose transporter‐4 (GLUT4) translocation was performed in surgically extracted muscle biopsies. All data are reported as median (25th/75th percentile) (trial registry: ISRCTN77569430 and ISRCTN19392591/ethics approval: Charité‐EA2/061/06 and Charité‐EA2/041/10).

Results

Fifty critically ill patients (admission SOFA 13) showed markedly decreased ISIs on Day 17 (interquartile range) 0.029 (0.022/0.048) (mg/min/kg)/(mU/L) compared with healthy controls 0.103 (0.087/0.111), P < 0.001. ISI correlated with muscle strength measured by medical research council (MRC) score at first awakening (r = 0.383, P = 0.026) and at ICU discharge (r = 0.503, P = 0.002). Different physiotherapeutic strategies showed no effect on the ISI [sPT 0.029 (0.019/0.053) (mg/min/kg)/(mU/L) vs. pPT 0.026 (0.023/0.041) (mg/min/kg)/(mU/L) vs. pPT+ 0.029 (0.023/0.042) (mg/min/kg)/(mU/L); P = 0.919]. Regardless of the physiotherapeutic strategy metabolic flexibility was reduced. Relative change of lactate/pyruvate ratio during HE clamp is as follows: sPT 0.09 (−0.13/0.27) vs. pPT 0.07 (−0.16/0.31) vs. pPT+ −0.06 (−0.19/0.16), P = 0.729, and relative change of glycerol concentration: sPT −0.39 (−0.8/−0.12) vs. pPT −0.21 (−0.33/0.07) vs. pPT+ −0.21 (−0.44/−0.03), P = 0.257. The majority of ICU patients showed abnormal localization of GLUT4 with membranous GLUT4 distribution in 37.5% (3 of 8) of ICU patients receiving sPT, in 42.9% (3 of 7) of ICU patients receiving pPT, and in 53.8% (7 of 13) of ICU patients receiving pPT+ (no statistical testing possible).

Conclusions

Our data suggest that a higher duration of muscle activating measures had no impact on insulin sensitivity or metabolic flexibility in critically ill patients with sepsis‐related multiple organ failure.
Uptake of community‐ versus clinic‐based antiretroviral therapy dispensing in the Central Chronic Medication Dispensing and Distribution program in South Africa
Authors: Bassett Ingrid V. , Yan Joyce , Govere Sabina , Khumalo Anele , Ngobese Nompumelelo , Shazi Zinhle , Nzuza Mpilonhle , Bunda Bridget A. , Wara Nafisa J. , Stuckwisch Ashley , Zionts Dani , Dube Nduduzo , Tshabalala Sandile , Bogart Laura M. , Parker Robert A.
Published: January 2022

Introduction

South Africa's government‐led Central Chronic Medication Dispensing and Distribution (CCMDD) program offers people living with HIV the option to collect antiretroviral therapy at their choice of community‐ or clinic‐based pick‐up points intended to increase convenience and decongest clinics. To understand CCMDD pick‐up point use among people living with HIV, we evaluated factors associated with uptake of a community‐ versus clinic‐based pick‐up point at CCMDD enrolment.

Methods

We collected baseline data from October 2018 to March 2020 on adults (≥18 years) who met CCMDD clinical eligibility criteria (non‐pregnant, on antiretroviral therapy for ≥1 year and virologically suppressed) as part of an observational cohort in seven public clinics in KwaZulu‐Natal. We identified factors associated with community‐based pick‐up point uptake and fit a multivariable logistic regression model, including age, gender, employment status, self‐perceived barriers to care, self‐efficacy, HIV‐related discrimination, and perceived benefits and challenges of CCMDD.

Results and Discussion

Among 1521 participants, 67% were females, with median age 36 years (IQR 30–44). Uptake of a community‐based pick‐up point was associated with younger age (aOR 1.18 per 10‐year decrease, 95% CI 1.05–1.33), being employed ≥40 hours per week (aOR 1.42, 95% CI 1.10–1.83) versus being unemployed, no self‐perceived barriers to care (aOR 1.42, 95% CI 1.09–1.86) and scoring between 36 and 39 (aOR 1.44, 95% CI 1.03–2.01) or 40 (aOR 1.91, 95% CI 1.39–2.63) versus 10–35 on the self‐efficacy scale, where higher scores indicate greater self‐efficacy. Additional factors included more convenient pick‐up point location (aOR 2.32, 95% CI 1.77–3.04) or hours (aOR 5.09, 95% CI 3.71–6.98) as perceived benefits of CCMDD, and lack of in‐clinic follow‐up after a missed collection date as a perceived challenge of CCMDD (aOR 4.37, 95% CI 2.30–8.31).

Conclusions

Uptake of community‐based pick‐up was associated with younger age, full‐time employment, and systemic and structural factors of living with HIV (no self‐perceived barriers to care and high self‐efficacy), as well as perceptions of CCMDD (convenient pick‐up point location and hours, lack of in‐clinic follow‐up). Strategies to facilitate community‐based pick‐up point uptake should be tailored to patients’ age, employment, self‐perceived barriers to care and self‐efficacy to maximize the impact of CCMDD in decongesting clinics.
Opposing roles for sMAdCAM and IL-15 in COVID-19 associated cellular immune pathology
Authors: Singh Amit Kumar , Kasarpalkar Nandini , Bhowmick Shilpa , Paradkar Gaurav , Talreja Mayur , Shah Karan , Tiwari Abhishek , Palav Harsha , Kaginkar Snehal , Kulkarni Rajiv , Patil Ashwini , Kalsurkar Varsha , Agrawal Sachee , Shastri Jayanthi , Dere Rajesh , Bharmal Ramesh , Mahale Smita D , Bhor Vikrant M , Patel Vainav
Published: January 2022

Immune cell dysregulation and lymphopenia characterize COVID-19 pathology in moderate to severe disease. While underlying inflammatory factors have been extensively studied, homeostatic and mucosal migratory signatures remain largely unexplored as causative factors. In this study, we evaluated the association of circulating IL-6, soluble mucosal addressin cell adhesion molecule (sMAdCAM), and IL-15 with cellular dysfunction characterizing mild and hypoxemic stages of COVID-19. A cohort of SARS-CoV-2 infected individuals (n = 130) at various stages of disease progression together with healthy controls (n = 16) were recruited from COVID Care Centres (CCCs) across Mumbai, India. Multiparametric flow cytometry was used to perform in-depth immune subset characterization and to measure plasma IL-6 levels. sMAdCAM, IL-15 levels were quantified using ELISA. Distinct depletion profiles, with relative sparing of CD8 effector memory and CD4+ regulatory T cells, were observed in hypoxemic disease within the lymphocyte compartment. An apparent increase in the frequency of intermediate monocytes characterized both mild as well as hypoxemic disease. IL-6 levels inversely correlated with those of sMAdCAM and both markers showed converse associations with observed lympho-depletion suggesting opposing roles in pathogenesis. Interestingly, IL-15, a key cytokine involved in lymphocyte activation and homeostasis, was detected in symptomatic individuals but not in healthy controls or asymptomatic cases. Further, plasma IL-15 levels negatively correlated with T, B, and NK count suggesting a compensatory production of this cytokine in response to the profound lymphopenia. Finally, higher levels of plasma IL-15 and IL-6, but not sMAdCAM, were associated with a longer duration of hospitalization.
Interaction mechanism of pelargonidin against tyrosinase by multi‐spectroscopy and molecular docking
Authors: Tao Yanzhou , Chen Rongda , Fan Yangyang , Liu Guiming , Wang Meizi , Wang Suqing , Li Li
Published: January 2022

The interaction mechanism of pelargonidin (PG) with tyrosinase was investigated by multi‐spectroscopy and molecular docking. As a result, PG had strong inhibitory activity on tyrosinase with the IC50 value of 41.94 × 10−6 mol·L−1. The inhibition type of PG against tyrosinase was determined as a mixed‐mode. Meanwhile, the fluorescence of tyrosinase was quenched statically by PG, and accompanied by non‐radiative energy transfer. The three‐dimensional (3‐D) fluorescence, ultraviolet‐visible spectroscopy (UV‐Vis) and circular dichroism spectroscopies (CD) indicated that PG decreased the hydrophobicity of the micro‐environment around tryptophan (Trp) and tyrosine (Tyr), which resulted in the conformational change of tyrosinase. In addition, fluorescence and molecular docking analysis indicated that PG bound to tyrosinase via hydrogen bonds (H‐bonds) and van der Waals force (vdW force). We herein recommended that PG might be a potential candidate drug for the treatment of melanin‐related diseases.
Inhibitory activities of Ranunculus japonicus Thunb. ethanol extract against hepatitis B virus
Authors: Luo Minhui , Chen Zhuohang , Liu Miaoya , Liang Qian , Han Ruilian , Liang Zongsuo , Ye Zhuoming , Liu Kuancheng
Published: January 2022

The chronic hepatitis B virus (HBV) infection is a worldwide public health problem, which cannot be cured by current therapeutics due to the persistence of viral CCC DNA in the infected hepatocytes. Screening from medicinal herbs for anti‐HBV activities showed that the ethanol extract from Ranunculus japonicus Thunb. could decrease the production of HBV e antigen (HBeAg). Further study showed that the extract had no effect on core protein expression but significantly reduced the efficiency of viral capsid assembly. The levels of viral pgRNA and total core DNA were not affected significantly. However, the ratio of RC DNA/SS DNA decreased, indicating that the conversion of RC DNA from SS DNA was delayed by the extract. More interestingly, though similar levels of RC DNA were accumulated, the CCC DNA level and its formation efficiency were reduced significantly, which was also consistent with the decreased level of HBeAg, indicating that R. japonicus Thunb. extract could inhibit the CCC DNA formation. Together, this study found that R. japonicus Thunb. extract could inhibit HBV replication at multiple steps, especially showed significant inhibitory effects on capsid assembly and CCC DNA formation.
The intersection of fracture healing and infection: Orthopaedics research society workshop 2021
Authors: Hellwinkel Justin E. , Working Zachary M , Certain Laura , García Andrés J. , Wenke Joseph C. , Bahney Chelsea S.
Published: January 2022

Infection is a common cause of impaired fracture healing. In the clinical setting, definitive fracture treatment and infection are often treated separately and sequentially, by different clinical specialties. The ability to treat infection while promoting fracture healing will greatly reduce the cost, number of procedures, and patient morbidity associated with infected fractures. In order to develop new therapies, scientists and engineers must understand the clinical need, current standards of care, pathologic effects of infection on fractures, available preclinical models, and novel technologies. One of the main causes of poor fracture healing is infection; unfortunately, bone regeneration and infection research are typically approached independently and viewed as two separate disciplines. Here, we aim to bring these two groups together in an educational workshop to promote research into the basic and translational science that will address the clinical challenge of delayed fracture healing due to infection. Statement of clinical significance: Infection and nonunion are each feared outcomes in fracture care, and infection is a significant driver of nonunion. The impact of nonunions on patie[Q2]nt well‐being is substantial. Outcome data suggests a long bone nonunion is as impactful on health‐related quality of life measures as a diagnosis of type 1 diabetes and fracture‐related infection has been shown to significantly l[Q3]ower a patient's quality of life for over 4 years.  Although they frequently are associated with one another, the treatment approaches for infections and nonunions are not always complimentary and cannot be performed simultaneously without accepting tradeoffs. Furthermore, different clinical specialties are often required to address the problem, the orthopedic surgeon treating the fracture and an infectious disease specialist addressing the sources of infection. A sequential approach that optimizes treatment parameters requires more time, more surgeries, and thus confers increased morbidity to the patient. The ability to solve fracture healing and infection clearance simultaneously in a contaminated defect would benefit both the patient and the health care system.
DL‐EPR and AFM study of sensitization of a 17%Cr multiphase stainless steel
Authors: Tavares Sérgio S. M. , de Sampaio Marcelo T. G. , Perez Geronimo , de Almeida Brígida B. , Ponzio Eduardo A.
Published: January 2022

The corrosion behavior of a 17Cr multiphase martensitic stainless steel quenched (1000°C, water cooling) and tempered at different temperatures was determined using the double‐loop electrochemical potentiokinetic reactivation (DL‐EPR) test. The specimen tempered at 550°C showed the highest degree of sensitization. For the specimen treated at 600°C, a healing process was observed, followed by a second sensitization at 625°C coincident with an increase in the austenite volume fraction. This phase was quantified using the magnetic method and an increase in this phase was observed with the tempering temperature. The DL‐EPR results were correlated with surface aspects obtained by atomic force microscopy.
Toward MR‐only proton therapy planning for pediatric brain tumors: Synthesis of relative proton stopping power images with multiple sequence MRI and development of an online quality assurance tool
Authors: Wang Chuang , Uh Jinsoo , Patni Tushar , Merchant Thomas , Li Yimei , Hua Chia‐ho , Acharya Sahaja
Published: January 2022

Purpose

To generate synthetic relative proton stopping power (sRPSP) images from magnetic resonance imaging (MRI) sequence(s) and develop an online quality assurance (QA) tool for sRPSP to facilitate safe integration of magnetic resonance (MR)‐only proton planning into clinical practice.

Materials and methods

Planning computed tomography (CT) and MR images of 195 pediatric brain tumor patients were utilized (training: 150, testing: 45). Seventeen consistent‐cycle generative adversarial network (ccGAN) models were trained separately using paired CT‐converted RPSP and MRI datasets to transform a subject's MRI into sRPSP. T1‐weighted (T1W), T2‐weighted (T2W), and FLAIR MRI were permutated to form 17 combinations, with or without preprocessing, for determining the optimal training sequence(s). For evaluation, sRPSP images were converted to synthetic CT (sCT) and compared to the real CT in terms of mean absolute error (MAE) in Hounsfield units (HU). For QA, sCT was deformed and compared to a reference template built from training dataset to produce a flag map, highlighting pixels that deviate by >100 HU and fall outside the mean ± standard deviation reference intensity. The gamma intensity analysis (10%/3 mm) of the deformed sCT against the QA template on the intensity difference was investigated as a surrogate of sCT accuracy.

Results

The sRPSP images generated from a single T1W or T2W sequence outperformed that generated from multi‐MRI sequences in terms of MAE (all p < 0.05). Preprocessing with N4 bias and histogram matching reduced MAE of T2W MRI‐based sCT (54 ± 21 HU vs. 42 ± 13 HU, p = 0.002). The gamma intensity analysis of sCT against the QA template was highly correlated with the MAE of sCT against the real CT in the testing cohort (r = ‐0.89 for T1W sCT; r = ‐0.93 for T2W sCT).

Conclusion

Accurate sRPSP images can be generated from T1W/T2W MRI for proton planning. A QA tool highlights regions of inaccuracy, flagging problematic cases unsuitable for clinical use.