bluuebunny/crossref_metadata_2025 huggingface dataset:
Possible Counterexample of the Riemann Hypothesis
Authors: Vega Frank
Published: January 2022

Under the assumption that the Riemann hypothesis is true, von Koch deduced the improved asymptotic formula $\theta(x) = x + O(\sqrt{x} \times \log^{2} x)$, where $\theta(x)$ is the Chebyshev function. On the contrary, we prove if there exists some real number $x \geq 10^{8}$ such that $\theta(x) > x + \frac{1}{\log \log \log x} \times \sqrt{x} \times \log^{2} x$, then the Riemann hypothesis should be false. Note that, the von Koch asymptotic formula uses the Big $O$ notation, where $f(x) = O(g(x))$ means that there exists a positive real number $M$ and a real number $y$, such that $|f(x)| \leq M \times g(x)$ for all $x \geq y$. However, no matter how big we get the real number $y \geq 10^{8}$, the another positive real number $M$ could always prevail over the value of $\frac{1}{\log \log \log x}$ for sufficiently large numbers $x \geq y$.
Enhanced Mechanical Properties of Polyvinyl Chloride-Based Wood–Plastic Composites With Pretreated Corn Stalk
Authors: Shen Tao , Li Minghui , Zhang Bo , Zhong Lingxia , Lin Xiran , Yang Pengpeng , Li Ming , Zhuang Wei , Zhu Chenjie , Ying Hanjie
Published: January 2022

Wood–plastic composites (WPCs) are a type of environmentally friendly materials widely used in daily life. This paper selected low-value biomass, corn stalk (CS), as the lignocellulosic resource for polyvinyl chloride (PVC)-based WPCs. To depict the relationship between lignocellulosic composition (cellulose, hemicellulose, and lignin) and mechanical performance of WPCs, pretreatments have been optimized to selective removal of lignin using an alkaline-EtOH stewing process and selective removal of hemicellulose using an acid stewing process. The αC sample, in which both lignin and hemicellulose were removed, shows the highest degree of crystallinity (72.60%) as estimated from X-ray diffraction analysis results and fibrous morphology with the highest aspect ratio as seen in scanning electron microscopy images. Compared with PVC/CS, PVC/αC gives a substantial increase in tensile strength and modulus by 37.21 and 21.66% and flexural strength and modulus by 29.98 and 34.88%, respectively. These improvements lie in the reinforcing effect of a fibrous structure and the improved interfacial compatibility as proven by scanning electron microscopy and dynamic mechanical analyzer results. Considering the extracted lignin and hemicellulose can be further developed to valuable biochemicals, the pretreatment to CS adds value to both WPC materials and biorefinery products.
Development of Neuronal Guidance Fibers for Stimulating Electrodes: Basic Construction and Delivery of a Growth Factor
Authors: Wille Inga , Harre Jennifer , Oehmichen Sarah , Lindemann Maren , Menzel Henning , Ehlert Nina , Lenarz Thomas , Warnecke Athanasia , Behrens Peter
Published: January 2022

State-of-the-art treatment for sensorineural hearing loss is based on electrical stimulation of residual spiral ganglion neurons (SGNs) with cochlear implants (CIs). Due to the anatomical gap between the electrode contacts of the CI and the residual afferent fibers of the SGNs, spatial spreading of the stimulation signal hampers focused neuronal stimulation. Also, the efficiency of a CI is limited because SGNs degenerate over time due to loss of trophic support. A promising option to close the anatomical gap is to install fibers as artificial nerve guidance structures on the surface of the implant and install on these fibers drug delivery systems releasing neuroprotective agents. Here, we describe the first steps in this direction. In the present study, suture yarns made of biodegradable polymers (polyglycolide/poly-ε-caprolactone) serve as the basic fiber material. In addition to the unmodified fiber, also fibers modified with amine groups were employed. Cell culture investigations with NIH 3T3 fibroblasts attested good cytocompatibility to both types of fibers. The fibers were then coated with the extracellular matrix component heparan sulfate (HS) as a biomimetic of the extracellular matrix. HS is known to bind, stabilize, modulate, and sustainably release growth factors. Here, we loaded the HS-carrying fibers with the brain-derived neurotrophic factor (BDNF) which is known to act neuroprotectively. Release of this neurotrophic factor from the fibers was followed over a period of 110 days. Cell culture investigations with spiral ganglion cells, using the supernatants from the release studies, showed that the BDNF delivered from the fibers drastically increased the survival rate of SGNs in vitro. Thus, biodegradable polymer fibers with attached HS and loaded with BDNF are suitable for the protection and support of SGNs. Moreover, they present a promising base material for the further development towards a future neuronal guiding scaffold.
IGF2BP2 Induces U251 Glioblastoma Cell Chemoresistance by Inhibiting FOXO1-Mediated PID1 Expression Through Stabilizing lncRNA DANCR
Authors: Han Junfei , Yu Xiaojun , Wang Shanxi , Wang Yingguang , Liu Qikun , Xu Haoran , Wang Xiaosong
Published: January 2022

Glioma is the most common type of malignant tumor of the nervous system and is characterized by high mortality and poor outcome. This study aims to investigate the mechanism underlying IGF2 mRNA-binding protein 2 (IGF2BP2) and long noncoding RNA DANCR in etoposide resistance of glioblastoma (GBM) cells. Bioinformatics analysis identified the IGF2BP2-related regulators and DANCR target genes, which were subsequently evaluated by RNA pull-down and RIP assays. We exposed GBM cells to etoposide and thus established etoposide-resistant cells. Through functional experiments, we evaluated the interrelationship among IGF2BP2, DANCR, phosphotyrosine interaction domain containing 1 (PID1), and forkhead box protein O1 (FOXO1) and further assessed their impact on the sensitivity of GBM cells to etoposide. IGF2BP2 and DANCR were highly expressed in glioma cells and tissues, whereas PID1 and FOXO1 were poorly expressed. Mechanistically, overexpression of IGF2BP2 promoted DANCR stability and reduced DANCR methylation, whereas silencing of IGF2BP2 reduced survival of GBM cells and etoposide-resistant cells. Besides, DANCR interacted with FOXO1 to promote the ubiquitination of FOXO1. FOXO1 promoted the transcriptional expression of PID1, enhancing the chemotherapy sensitivity of GBM cells, but overexpression of PID1 reversed the impact of IGF2BP2. Collectively, IGF2BP2 inhibits PID1 expression through the DANCR/FOXO1 axis, inducing drug resistance in GBM cells, and promoting glioma progression.
A Prognostic Risk Score Based on Hypoxia-, Immunity-, and Epithelialto-Mesenchymal Transition-Related Genes for the Prognosis and Immunotherapy Response of Lung Adenocarcinoma
Authors: Ouyang Wenhao , Jiang Yupeng , Bu Shiyi , Tang Tiantian , Huang Linjie , Chen Ming , Tan Yujie , Ou Qiyun , Mao Luhui , Mai Yingjie , Yao Herui , Yu Yunfang , Lin Xiaoling
Published: January 2022

Background: Lung adenocarcinoma (LUAD), the most common subtype of non-small cell lung cancer (NSCLC), is associated with poor prognosis. However, current stage-based clinical methods are insufficient for survival prediction and decision-making. This study aimed to establish a novel model for evaluating the risk of LUAD based on hypoxia, immunity, and epithelial-mesenchymal transition (EMT) gene signatures.

Methods: In this study, we used data from TCGA-LUAD for the training cohort and GSE68465 and GSE72094 for the validation cohorts. Immunotherapy datasets GSE135222, GSE126044, and IMvigor210 were obtained from a previous study. Using bioinformatic and machine algorithms, we established a risk model based on hypoxia, immune, and EMT gene signatures, which was then used to divide patients into the high and low risk groups. We analyzed differences in enriched pathways between the two groups, following which we investigated whether the risk score was correlated with stemness scores, genes related to m6A, m5C, m1A and m7G modification, the immune microenvironment, immunotherapy response, and multiple anti-cancer drug sensitivity.

Results: Overall survival differed significantly between the high-risk and low-risk groups (HR = 4.26). The AUCs for predicting 1-, 3-, and 5-year survival were 0.763, 0.766, and 0.728, respectively. In the GSE68465 dataset, the HR was 2.03, while the AUCs for predicting 1-, 3-, and 5-year survival were 0.69, 0.651, and 0.618, respectively. The corresponding values in the GSE72094 dataset were an HR of 2.36 and AUCs of 0.653, 0.662, and 0.749, respectively. The risk score model could independently predict OS in patients with LUAD, and highly correlated with stemness scores and numerous m6A, m5C, m1A and m7G modification-related genes. Furthermore, the risk model was significantly correlated with multiple immune microenvironment characteristics. In the GSE135222 dataset, the HR was 4.26 and the AUC was 0.702. Evaluation of the GSE126044 and IMvigor210 cohorts indicated that PD-1/PD-LI inhibitor treatment may be indicated in patients with low risk scores, while anti-cancer therapy with various drugs may be indicated in patients with high risk scores.

Conclusion: Our novel risk model developed based on hypoxia, immune, and EMT gene signatures can aid in predicting clinical prognosis and guiding treatment in patients with LUAD.
USP25 Expression in Peripheral Blood Mononuclear Cells Is Associated With Bone Mineral Density in Women
Authors: Shen Jianlin , Fu Bowen , Wu Yanjiao , Yang Yang , Lin Xiaoning , Lin Haibin , Liu Huan , Huang Wenhua
Published: January 2022

Osteoporosis is the most common metabolic bone disease in postmenopausal women. As precursors of osteoclasts, peripheral blood mononuclear cells are accessible and considered suitable models for studying osteoporosis pathology. Ubiquitination is a crucial protein degradation system in bone metabolism. The aim of this study was to identify potential ubiquitination-related genes in PBMCs that are related to osteoporosis pathogenesis. Therefore, we performed an integrated analysis of osteoporosis-related microarray datasets. With the obtained ubiquitination-related gene set, weighted gene coexpression network analysis was performed. The results showed that genes in the turquoise module were correlated with menopause, and 48 genes were identified as hub genes. A differential expression analysis revealed 43 differentially expressed genes between pre- and postmenopausal samples. After integrating the information on differentially expressed menopause-related genes, we found that several members of the ubiquitin-specific protease (USP) family (USP1, USP7, USP9X, USP16, and USP25) were highly expressed in samples from postmenopausal female and that, USP25 expression was significantly higher in low-BMD samples than in high-BMD samples among samples from premenopausal subjects (p = 0.0013) and among all samples (p = 0.013). Finally, we verified the protein expression of USP25 in PBMCs by performing Western blot analysis, which yielded results consistent with the aforementioned results. Moreover, by assessing GTEx datasets, we found that USP25 expression was highly correlated with TRAF6 expression in whole blood (p < 0.001). We also tested the protein expression levels of TRAF6 in PBMCs and found that it was positively correlated with USP25 expression (p = 0.036). Our results reveal that the ubiquitin-specific protease family may play important roles in menopause and that USP25 is related to osteoporosis pathogenesis.
Cation-Chloride Cotransporters, Na/K Pump, and Channels in Cell Water/Ionic Balance Regulation Under Hyperosmolar Conditions: In Silico and Experimental Studies of Opposite RVI and AVD Responses of U937 Cells to Hyperosmolar Media
Authors: Yurinskaya Valentina E. , Vereninov Alexey A.
Published: January 2022

Studying the transport of monovalent ions across the cell membrane in living cells is complicated by the strong interdependence of fluxes through parallel pathways and requires therefore computational analysis of the entire electrochemical system of the cell. Current paper shows how to calculate changes in the cell water balance and ion fluxes caused by changes in the membrane channels and transporters during a normal regulatory increase in cell volume in response to osmotic cell shrinkage (RVI) followed by a decrease in cell volume associated with apoptosis (AVD). Our recently developed software is used as a computational analysis tool and the established human lymphoid cells U937 are taken as an example of proliferating animal cells. It is found that, in contrast to countless statements in the literature that cell volume restoration requires the activation of certain ion channels and transporters, the cellular responses such as RVI and AVD can occur in an electrochemical system like U937 cells without any changes in the state of membrane channels or transporters. These responses depend on the types of chloride cotransporters in the membrane and differ in a hyperosmolar medium with additional sucrose and in a medium with additional NaCl. This finding is essential for the identification of the true changes in membrane channels and transporters responsible for RVI and AVD in living cells. It is determined which changes in membrane parameters predicted by computational analysis are consistent with experimental data obtained on living human lymphoid cells U937, Jurkat, and K562 and which are not. An essential part of the results is the developed software that allows researchers without programming experience to calculate the fluxes of monovalent ions via the main transmembrane pathways and electrochemical gradients that move ions across the membrane. The software is available for download. It is useful for studying the functional expression of the channels and transporters in living cells and understanding how the cell electrochemical system works.
IR780 Based Sonotherapeutic Nanoparticles to Combat Multidrug-Resistant Bacterial Infections
Authors: Huang Biying , Wang Long , Tang Kui , Chen Sijie , Xu Yan , Liao Haiqin , Niu Chengcheng
Published: January 2022

Multidrug-resistant (MDR) bacterial strains have emerged and weakened the therapeutic effects of antibacterial drugs. Sonodynamic therapy (SDT) takes advantage of noninvasiveness and deep tissue-penetrating features and has been rejuvenated to combat MDR bacteria and their biofilm-associated infections. To improve the efficacy of antibacterial SDT, we first developed IR780-based PLGA nanoparticles as sonosensitizers for high-frequency ultrasound (US)-monitored antibacterial SDT of MRSA myositis by therapeutic low-frequency US. In this study, the developed shell-core-structured IR780@PLGA nanoparticles were designed with a polymer shell PLGA with the sonosensitizer IR780 loaded on. High-frequency diagnostic US was introduced to monitor the sonotherapeutic progression of bacterial myositis by therapeutic low-frequency US. Importantly, the in vitro and in vivo results confirmed that IR780@PLGA nanoparticles combined with US irradiation possess high efficiency for antibacterial therapy. This approach provides a simple and efficient strategy to monitor and combat MDR bacterial infection.
Epstein-Barr Virus-Associated Smooth Muscle Tumor and Its Correlation With CD4 Levels in a Patient With HIV Infection
Authors: Chong Yoon Bin , Lu Po-Liang , Ma Yu-Chun , Yin Hsin-Ling , Chang Chih-Hui
Published: January 2022

Epstein-Barr virus-associated smooth muscle tumor (EBV-SMT) is a rare tumor found in immunocompromised patients, and its treatment is not well-established. A role for antiretroviral therapy in human immunodeficiency virus (HIV)-related EBV-SMT has been proposed; however, the relevance of tumor size, CD4 levels, and immune reconstitution inflammatory syndrome (IRIS) has not been previously reported. We present the first case, to our knowledge, of a tumor that shrank in association with elevated CD4 counts. IRIS occurred in this case following antiretroviral therapy. This finding highlights the importance of the immune response in HIV-related EBV-SMT.
Reassessing the Role of Anthropogenic Climate Change in the Extinction of Silphium
Authors: Pollaro Paul , Robertson Paul
Published: January 2022

The famed ancient herb, known to the Romans as silphium (Greek silphion), is widely regarded as the first recorded instance of human-induced species extinction. Modern scholars have largely credited direct exploitation (e.g., over-harvesting; over-grazing) as the primary cause of silphium's extinction, due to an overwhelming demand for the plant in ancient times. Recent research has revealed strict cold-stratification requirements for the germination of silphium's closest living relatives, revealing the likelihood that silphium shared these same germination requirements. Documented environmental changes in ancient Cyrenaica (e.g., widespread deforestation; cropland expansion) likely resulted in accelerated rates of desertification throughout the region as well as the direct disturbance of silphium's habitat, effectively eliminating the necessary conditions for silphium's successful germination and growth within its native range. Contrary to previous conclusions, this evidence suggests that anthropogenic environmental change was instead the dominant factor in silphium's extinction, marking silphium as the first recorded instance of human-induced climate-based extinction.
Biodiversity Exploitation for Online Entertainment
Authors: Khelifa Rassim , Mellal Mohammed Khalil , Mahdjoub Hayat , Hasanah Nur , Kremen Claire
Published: January 2022

Anthropogenic wildlife exploitation threatens biodiversity worldwide. With the emergence of online trading which facilitates the physical movement of wildlife across countries and continents, wildlife conservation is more challenging than ever. One form of wildlife exploitation involves no physical movement of organisms, presenting new challenges. It consists of hunting and fishing “experiments” for monetized online entertainment. Here we analyze >200 online videos of these so-called experiments in the world's largest video platform (YouTube). These videos generated about half a billion views between 2019 and 2020. The number of target species (including threatened animals), videos, and views increased rapidly during this period. The material used in these experiments raises serious ethical questions about animal welfare and the normalization of violence to animals on the Internet. The emergence of this phenomenon highlights the need for online restriction of this type of content to limit the spread of animal cruelty and the damage to global biodiversity. It also sheds light on some conservation gaps in the virtual sphere of the Internet which offers biodiversity-related business models that has the potential to spread globally.
Case Report: COVID-19 Vaccination Associated Fulminant Myocarditis
Authors: Cui Guanglin , Li Rui , Zhao Chunxia , Wang Dao Wen
Published: January 2022

Herein, we describe a novel finding of fulminant myocarditis (FM) in two subjects the day after administration of the first dose of the currently available inactivated SARS-CoV-2 vaccine (Vero cell). Cardiac magnetic resonance imaging revealed extensive myocardial edema and necrosis. A pathologic evaluation of the endocardial biopsy tissues revealed inflammatory cell (lymphocytes) infiltration and interstitial edema, myocyte necrosis, and focal areas of fibrosis. A life-support-based comprehensive treatment regimen comprising mechanical circulatory support using intra-aortic balloon pulsation and immunomodulatory therapy—glucocorticoids and intravenous immunoglobulin—was used to treat the patients with FM; eventually, the patients recovered and were discharged. To our knowledge, these are the first two reported cases of FM, with no other identified cause or associated illness, after receiving the inactivated SARS-CoV-2 vaccine (Vero cell). These findings suggest a novel pathogenesis of myocarditis which mentions to pay more attention to this rare, but lethal complication of COVID-19 vaccination.
Identification of Rare Variants in Right Ventricular Outflow Tract Obstruction Congenital Heart Disease by Whole-Exome Sequencing
Authors: Zhou Yue , Bai Kai , Wang Yu , Meng Zhuo , Zhou Shuang , Jiang Shiwei , Wang Hualin , Wang Jian , Yang Mei , Wang Qingjie , Sun Kun , Chen Sun
Published: January 2022

Background

Pulmonary atresia (PA) is a kind of congenital heart disease characterized by right ventricular outflow tract obstruction. It is divided into PA with intact ventricular septum (PA/IVS) whose favorable form is pulmonary valvular stenosis (PS), and PA with ventricular septal defect (PA/VSD) whose favorable form is tetralogy of Fallot (TOF). Due to limitations in genetics etiology, whole-exome sequencing (WES) was utilized to identify new variants associated with the diseases.

Methods

The data from PS-PA/IVS (n = 74), TOF-PA/VSD (n = 100), and 100 controls were obtained. The common sites between PS and PA/IVS, PA/VSD and TOF, were compared. The novel rare damage variants, and candidate genes were identified by gene-based burden analysis. Finally, the enrichment analysis of differential genes was conducted between case and control groups.

Results

Seventeen rare damage variants located in seven genes were predicted to be associated with the PS through burden analysis. Enrichment analysis identified that the Wnt and cadherin signaling pathways were relevant to PS-PA/IVS.

Conclusion

This study put forth seven candidate genes (APC, PPP1R12A, PCK2, SOS2, TNR, MED13, and TIAM1), resulting in PS-PA/IVS. The Wnt and cadherin signaling pathways were identified to be related to PS-PA/IVS by enrichment analysis. This study provides new evidence for exploring the genetic mechanism of PS-PA/IVS.
Construction and Analysis of the lncRNA-miRNA-mRNA Network Based on Competing Endogenous RNA in Atrial Fibrillation
Authors: Ke Xiangyu , Zhang Junguo , Huang Xin , Li Shuai , Leng Meifang , Ye Zebing , Li Guowei
Published: January 2022

Background

Accumulated studies have revealed that long non-coding RNAs (lncRNAs) play critical roles in human diseases by acting as competing endogenous RNAs (ceRNAs). However, functional roles and regulatory mechanisms of lncRNA-mediated ceRNA in atrial fibrillation (AF) remain unknown. In the present study, we aimed to construct the lncRNA-miRNA-mRNA network based on ceRNA theory in AF by using bioinformatic analyses of public datasets.

Methods

Microarray data sets of GSE115574 and GSE79768 from the Gene Expression Omnibus database were downloaded. Twenty-one AF right atrial appendage (RAA) samples and 22 sinus rhythm (SR) subjects RAA samples were selected for subsequent analyses. After merging all microarray data and adjusting for batch effect, differentially expressed genes were identified. Gene Ontology (GO) categories and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were carried out. A ceRNA network was constructed.

Result

A total of 8 lncRNAs and 43 mRNAs were significantly differentially expressed with fold change >1.5 (p < 0.05) in RAA samples of AF patients when compared with SR. GO and KEGG pathway analysis showed that cardiac muscle contraction pathway were involved in AF development. The ceRNA was predicted by co-expressing LOC101928304/ LRRC2 from the constructional network analysis, which was competitively combined with miR-490-3p. The expression of LOC101928304 and LRRC were up-regulated in myocardial tissue of patients with AF, while miR-490-3p was down-regulated.

Conclusion

We constructed the LOC101928304/miR-490-3p/LRRC2 network based on ceRNA theory in AF in the bioinformatic analyses of public datasets. The ceRNA network found from this study may help improve our understanding of lncRNA-mediated ceRNA regulatory mechanisms in the pathogenesis of AF.
COVID-19 Accelerating Academic Teachers’ Digital Competence in Distance Teaching
Authors: Myyry Liisa , Kallunki Veera , Katajavuori Nina , Repo Saara , Tuononen Tarja , Anttila Henrika , Kinnunen Päivi , Haarala-Muhonen Anne , Pyörälä Eeva
Published: January 2022

This study examines, using a cross-sectional approach, the digital competence of academic teachers at a time when teaching shifted to digital distance learning at the start of the COVID-19 pandemic. Teachers from different academic fields at a large multidisciplinary Finnish university (N = 265) responded to a questionnaire about the purposes for which they use digital tools in teaching, how they evaluated their competence at distance teaching during the lockdown of March-May 2020 and their beliefs about distance teaching. The respondents used digital tools in teaching mostly for delivering information. According to their evaluations, their competence in distance teaching increased during the early stages of COVID-19 pandemic in 2020, but their beliefs about distance teaching did not relate to the feelings of competence. Respondents with no experience in distance teaching before the lockdown evaluated their competence as having increased more than did respondents with previous experience. The implications of the findings for understanding competence development are then discussed.
From Face-To-Face to the Online Space: The Continued Relevance of Connecting Students With Each Other and Their Learning Post COVID-19
Authors: Garrad Traci-Ann , Page Angela
Published: January 2022

At the time of writing, the largest state in Australia is once more in full lockdown because of surging cases from the new variant strain of COVID-19. During the last lockdown in early 2020, we conducted a study analyzing the efficacy of mapping best-practice face-to-face university teaching into the online space. This article reports on the results of a survey from the perspective of student belonging. Isolation was the most prevalent theme recorded by students despite not being one of the research questions asked. The importance of adopting the model in online university courses in the current/post-COVID-19 world is presented.
Perspectives on Fully Synthesized Sustainable Aviation Fuels: Direction and Opportunities
Authors: Kramer Stephen , Andac Gurhan , Heyne Joshua , Ellsworth Joseph , Herzig Peter , Lewis Kristin C.
Published: January 2022

The aviation sector seeks to reduce greenhouse gas (GHG) emissions, with manufacturers and airlines announcing “zero-emission” goals and plans. Reduced carbon aviation fuels are central to meeting these goals. However, current and near-term aircraft, which will remain flying for decades, are designed around the combustion of petroleum-based aviation kerosene (e.g., Jet A/A-1). Therefore, the industry has focused on the qualification and approval of synthesized (e.g., non-petroleum-based) aviation fuel components with maximum blend limit percentages to avoid the blended fuel having properties outside the accepted ranges for Jet A/A-1. The synthesized components approved for blending are not necessarily interchangeable with Jet A/A-1. They may lack certain required chemical components, such as aromatics, or may have other characteristics outside the allowable ranges. To ensure safety, these synthesized aviation fuel components are only qualified to be used in commercial aviation when blended up to approved limits. The sector seeks to move toward the capability of using 100% synthesized aviation fuels that also meet sustainability criteria, known as sustainable aviation fuels, or SAF. However, these fuels must be developed, assessed, and deployed appropriately. This paper explores key questions relating to the introduction of 100% SAF, concluding that:

• Near-term unblended synthesized aviation fuels must be “drop-in,” meaning they are compatible with existing aircraft and infrastructure.

• Stand-alone complete fuels could be qualified within 1–2 years, with blends of blending components to reach 100% synthesized fuels to follow.

• Sustainability criteria, while critical to sector acceptance, will continue to be assessed separately from technical performance.
Contributions of Ultrastructural Studies to the Knowledge of Filamentous Fungi Biology and Fungi-Plant Interactions
Authors: Faoro Franco , Faccio Antonella , Balestrini Raffaella
Published: January 2022

Since the first experiments in 1950s, transmission electron microscopy (TEM) observations of filamentous fungi have contributed extensively to understand their structure and to reveal the mechanisms of apical growth. Additionally, also in combination with the use of affinity techniques (such as the gold complexes), several aspects of plant-fungal interactions were elucidated. Nowadays, after the huge of information obtained from -omics techniques, TEM studies and ultrastructural observations offer the possibility to support these data, considering that the full comprehension of the mechanisms at the basis of fungal morphogenesis and the interaction with other organisms is closely related to a detailed knowledge of the structural features. Here, the contribution of these approaches on fungal biology is illustrated, focusing both on hyphae cell ultrastructure and infection structures of pathogenic and mycorrhizal fungi. Moreover, a concise appendix of methods conventionally used for the study of fungal ultrastructure is provided.
Advances in Delivery Mechanisms of CRISPR Gene-Editing Reagents in Plants
Authors: Laforest Larissa C. , Nadakuduti Satya Swathi
Published: January 2022

Gene-editing by CRISPR/Cas systems has revolutionized plant biology by serving as a functional genomics tool. It has tremendously advanced plant breeding and crop improvement by accelerating the development of improved cultivars, creating genetic variability, and aiding in domestication of wild and orphan crops. Gene-editing is a rapidly evolving field. Several advancements include development of different Cas effectors with increased target range, efficacy, and enhanced capacity for precise DNA modifications with base editing and prime editing. The existing toolbox of various CRISPR reagents facilitate gene knockouts, targeted gene insertions, precise base substitutions, and multiplexing. However, the major challenge in plant genome-editing remains the efficient delivery of these reagents into plant cells. Plants have larger and more complex genome structures compared to other living systems due to the common occurrence of polyploidy and other genome re-arrangements. Further, rigid cell walls surrounding plant cells deter the entry of any foreign biomolecules. Unfortunately, genetic transformation to deliver gene-editing reagents has been established only in a limited number of plant species. Recently, there has been significant progress in CRISPR reagents delivery in plants. This review focuses on exploring these delivery mechanisms categorized into Agrobacterium-mediated delivery and breakthroughs, particle bombardment-based delivery of biomolecules and recent improvements, and protoplasts, a versatile system for gene-editing and regeneration in plants. The ultimate goal in plant gene-editing is to establish highly efficient and genotype-independent reagent delivery mechanisms for editing multiple targets simultaneously and achieve DNA-free gene-edited plants at scale.
Omicron: A Heavily Mutated SARS-CoV-2 Variant Exhibits Stronger Binding to ACE2 and Potently Escapes Approved COVID-19 Therapeutic Antibodies
Authors: Shah Masaud , Woo Hyun Goo
Published: January 2022

The new SARS-CoV-2 variant of concern “Omicron” was recently spotted in South Africa and spread quickly around the world due to its enhanced transmissibility. The variant became conspicuous as it harbors more than 30 mutations in the Spike protein with 15 mutations in the receptor-binding domain (RBD) alone, potentially dampening the potency of therapeutic antibodies and enhancing the ACE2 binding. More worrying, Omicron infections have been reported in vaccinees in South Africa and Hong Kong, and that post-vaccination sera poorly neutralize the new variant. Here, we investigated the binding strength of Omicron with ACE2 and monoclonal antibodies that are either approved by the FDA for COVID-19 therapy or undergoing phase III clinical trials. Computational mutagenesis and free energy perturbation could confirm that Omicron RBD binds ACE2 ~2.5 times stronger than prototype SARS-CoV-2. Notably, three substitutions, i.e., T478K, Q493K, and Q498R, significantly contribute to the binding energies and almost doubled the electrostatic potential (ELE) of the RBDOmic–ACE2 complex. Omicron also harbors E484A substitution instead of the E484K that helped neutralization escape of Beta, Gamma, and Mu variants. Together, T478K, Q493K, Q498R, and E484A substitutions contribute to a significant drop in the ELE between RBDOmic–mAbs, particularly in etesevimab, bamlanivimab, and CT-p59. AZD1061 showed a slight drop in ELE and sotrovimab that binds a conserved epitope on the RBD; therefore, it could be used as a cocktail therapy in Omicron-driven COVID-19. In conclusion, we suggest that the Spike mutations prudently devised by the virus facilitate the receptor binding, weakening the mAbs binding to escape the immune response.
The Prognostic Value of FoxP3+ Tumour-Infiltrating Lymphocytes in Rectal Cancer Depends on Immune Phenotypes Defined by CD8+ Cytotoxic T Cell Density
Authors: Schnellhardt Sören , Hirneth Johannes , Büttner-Herold Maike , Daniel Christoph , Haderlein Marlen , Hartmann Arndt , Fietkau Rainer , Distel Luitpold
Published: January 2022

Tumour-infiltrating FoxP3+ regulatory T cells have been identified as both positive and negative prognostic factors in colorectal cancer (CRC) and rectal cancer (RC). In this study we investigated whether immune phenotypes, defined by CD8+ cytotoxic T cell density, may influence the prognostic association of FoxP3+ T cell densities in RC. Tissue microarrays from 154 rectal cancer resections were immunohistochemically double stained for CD8 and FoxP3. CD8+ and FoxP3+ cell densities were measured in the stromal and intraepithelial compartment. Stromal FoxP3+ cell densities were not associated with 10-year overall survival (OS). In the “immune-desert” phenotype, defined by very low stromal CD8+ cell density, a high density of stromal FoxP3+ T cells displayed a tendency towards an association with decreased 10-year OS (p = 0.179). In “inflamed” tumours, defined by high intraepithelial CD8+ T cell infiltration, the opposite was the case and high stromal FoxP3+ T cell densities were a positive prognostic factor (p = 0.048). Additionally, patients with an increased FoxP3/CD8 cell density ratio demonstrated a strong trend towards decreased 10-year OS (p = 0.066). These contrasting findings suggest functional heterogeneity within the group of FoxP3+ T cells. They are consistent with experimental studies which reported suppressive and non-suppressive populations of FoxP3+ T cells in CRC. Furthermore, our study demonstrates that CD8 immunohistochemistry may act as an instrument to identify tumours infiltrated by possibly functionally differing FoxP3+ T cell subtypes.
Immunological Resistance of Pseudosuccinea columella Snails From Cuba to Fasciola hepatica (Trematoda) Infection: What We Know and Where We Go on Comparative Molecular and Mechanistic Immunobiology, Ecology and Evolution
Authors: Alba Annia , Vázquez Antonio A. , Sánchez Jorge , Gourbal Benjamin
Published: January 2022

One of the most interesting biological models is that of snail–trematode interactions, many of which ultimately result in the transmission of several important diseases, particularly in the tropics. Herein, we review the scientific advances on a trematode–snail system in which certain populations of Pseudosuccinea columella (a common host species for trematodes) have been demonstrated naturally-resistant to Fasciola hepatica, in association with an effective encapsulation of the parasite by innate immune cells of the host, the hemocytes. Emphasis is made on the molecular and immunological features characterizing each P. columella phenotype in relation to their anti-parasitic competence, their distinctive ecological patterns and the existence of a significant cost of resistance. An integrative overview of the resistance to F. hepatica through comparative immunobiology, genetics and ecology is presented to hypothesize on the possible origins and evolution of this phenomenon and to postulate significant roles for parasite mediated-selection and environmental factors in shaping and maintaining the resistant phenotype in the field. Lastly, clues into future experimental perspectives to deeply characterize the interplay between P. columella and F. hepatica and the immunobiology of the resistance are also included. The advances revised in the present paper are only beginning to unravel mechanisms of anti-parasite innate defense responses and their evolutionary bases, and can facilitate the development of prospective approaches towards practical applications of P. columella resistance.
Autocrine/Paracrine Loop Between SCF+/c-Kit+ Mast Cells Promotes Cutaneous Melanoma Progression
Authors: Annese Tiziana , Tamma Roberto , Bozza Mariella , Zito Alfredo , Ribatti Domenico
Published: January 2022

c-Kit, or mast/stem cell growth factor receptor Kit, is a tyrosine kinase receptor structurally analogous to the colony-stimulating factor-1 (CSF-1) and platelet-derived growth factor (PDGF) CSF-1/PDGF receptor Tyr-subfamily. It binds the cytokine KITLG/SCF to regulate cell survival and proliferation, hematopoiesis, stem cell maintenance, gametogenesis, mast cell development, migration and function, and it plays an essential role in melanogenesis. SCF and c-Kit are biologically active as membrane-bound and soluble forms. They can be expressed by tumor cells and cells of the microenvironment playing a crucial role in tumor development, progression, and relapses. To date, few investigations have concerned the role of SCF+/c-Kit+ mast cells in normal, premalignant, and malignant skin lesions that resemble steps of malignant melanoma progression. In this study, by immunolabeling reactions, we demonstrated that in melanoma lesions, SCF and c-Kit were expressed in mast cells and released by themselves, suggesting an autocrine/paracrine loop might be implicated in regulatory mechanisms of neoangiogenesis and tumor progression in human melanoma.
RNA-Seq of Single Fish Cells – Seeking Out the Leukocytes Mediating Immunity in Teleost Fishes
Authors: Chan Justin T. H. , Kadri Safwen , Köllner Bernd , Rebl Alexander , Korytář Tomáš
Published: January 2022

The immune system is a complex and sophisticated biological system, spanning multiple levels of complexity, from the molecular level to that of tissue. Our current understanding of its function and complexity, of the heterogeneity of leukocytes, is a result of decades of concentrated efforts to delineate cellular markers using conventional methods of antibody screening and antigen identification. In mammalian models, this led to in-depth understanding of individual leukocyte subsets, their phenotypes, and their roles in health and disease. The field was further propelled forward by the development of single-cell (sc) RNA-seq technologies, offering an even broader and more integrated view of how cells work together to generate a particular response. Consequently, the adoption of scRNA-seq revealed the unexpected plasticity and heterogeneity of leukocyte populations and shifted several long-standing paradigms of immunology. This review article highlights the unprecedented opportunities offered by scRNA-seq technology to unveil the individual contributions of leukocyte subsets and their crosstalk in generating the overall immune responses in bony fishes. Single-cell transcriptomics allow identifying unseen relationships, and formulating novel hypotheses tailored for teleost species, without the need to rely on the limited number of fish-specific antibodies and pre-selected markers. Several recent studies on single-cell transcriptomes of fish have already identified previously unnoticed expression signatures and provided astonishing insights into the diversity of teleost leukocytes and the evolution of vertebrate immunity. Without a doubt, scRNA-seq in tandem with bioinformatics tools and state-of-the-art methods, will facilitate studying the teleost immune system by not only defining key markers, but also teaching us about lymphoid tissue organization, development/differentiation, cell-cell interactions, antigen receptor repertoires, states of health and disease, all across time and space in fishes. These advances will invite more researchers to develop the tools necessary to explore the immunology of fishes, which remain non-conventional animal models from which we have much to learn.
The Paradox of an Unpolluted Coastal Site Facing a Chronically Contaminated Industrial Area
Authors: Corinaldesi Cinzia , Bianchelli Silvia , Rastelli Eugenio , Varrella Stefano , Canensi Sara , Gambi Cristina , Lo Martire Marco , Musco Luigi , Bertocci Iacopo , Fanelli Emanuela , Lucia Giulia , Simoncini Nicola , Dell’Anno Antonio
Published: January 2022

Present and past industrial activities in coastal areas have left us a legacy of contamination and habitat degradation with potential implications for human health. Here, we investigated a coastal marine area enclosed in a Site of National Interest (SNI) of the central-western Adriatic (Mediterranean Sea), where priority actions of environmental remediation are required by governmental laws due the high environmental and human risk, and that is off-limits to any human activity since 2002. In particular, our investigation was focused on an area located in front of a chemical industry dismissed more than 3 decades ago. We report that the concentrations of heavy-metal and organic contaminants in the investigated sediments were generally lower than those expected to induce detrimental biological effects. Meiofaunal abundance, biomass and community structure changed among stations, but regardless of the distance from the abandoned industrial plant. Taxa richness within the SNI did not change significantly compared to the controls and the lack of some taxa in the SNI transects was not due to the contamination of the SNI area. The results of this study suggest a natural recovery of the marine area over 2 decades of restrictions on human activities, including fishing and shipping bans. If the hypothesis of the natural recovery of this SNI will be further confirmed by other studies, the plans for the identification and monitoring of the most polluted areas in Italy should necessarily be redefined also in the light of the Water Framework, the Marine Strategy Framework and the Environmental Quality Standard Directives.
Experimental Study on Residual Compressive Strength of Recycled Aggregate Concrete Under Fatigue Loading
Authors: You Fan , Luo Surong , Zheng Jianlan
Published: January 2022

Using recycled aggregate for producing concrete is an effective way to realize the resource utilization of construction and demolition waste. Understanding the variation law of the residual compressive strength of recycled aggregate concrete (RAC) under fatigue loading is significant for its practical application under repeated loads. Thus, this paper conducted uniaxial compression fatigue and static-load tests on recycled aggregate concrete with the different replacement ratios of recycled coarse aggregate, analyzed the probability distributions of the fatigue life and residual compressive strength of RAC and natural aggregate concrete (NAC), and revealed the attenuation law of their residual compressive strength. In addition, it studied and compared the evolution of fatigue damage to NAC and RAC by defining a damage variable based on residual compressive strength. The results show that the fatigue life of the RAC and NAC matches the two-parameter Weibull distribution, and their residual compressive strength follows the normal distribution. The attenuation law of the residual compressive strength of the RAC is similar to that of the NAC. In the initial stages of the fatigue loading, the attenuation is small, and the attenuation rate of the residual compressive strength of the RAC and NAC is low. In contrast, it gradually accelerates in the later stages; a sudden and significant decrease in the concrete strength is noticed in the final stage near failure. Compared with the NAC, the damage to the RAC develops faster in the initial stages of fatigue loading and slower in the later stages.
Efficacy and Safety of Topical Therapy With Botanical Products for Melasma: A Systematic Review and Meta-Analysis of Randomized Controlled Trials
Authors: Wang Tianyun , Wang Youmei , Wang Jue , Chen Hongwei , Qu Biao , Li Zheng
Published: January 2022

Botanical products have been increasingly popular in topical therapies for melasma, as presumed safer and milder than fully synthetic products. Although the efficacy of different topical botanicals has recently been substantiated through randomized controlled trials (RCTs), there is a lack of sufficiently pooled evidence on their efficacy and safety for the treatment of melasma. Herein, a systematic review and meta-analysis was conducted on the efficacy and safety of topical botanical products for the treatment of melasma, following Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA). All RCTs on the use of topical botanical products for the treatment of melasma in humans were included, except for trials enrolling pregnant patients. The primary outcome was Melasma Area and Severity Index (MASI) or its variation. The secondary outcomes included Mexameter® reading, melasma improvement evaluated by participants, and any reported adverse events (AEs). As a result, twelve eligible trials comprising 695 patients with melasma from 6 different countries were included. The topical botanical products contained active ingredients which varied among trials as follows: herb-derived molecule, extracts of a single herb, and extracts of compound herbs. Topical therapy with botanical products significantly improved melasma with a large effect on MASI reduction (SMD −0.79, 95% CI −1.14 to −0.44, p < 0.00001), and a moderate effect on Mexameter® reading reduction (SMD −0.52, 95% CI −0.81 to 0.23, p = 0.0005), when compared with placebo. It also showed a similar improvement of melasma with a better safety profile (RR 0.37, 95% CI 0.15–0.88, p = 0.02), when compared with active-comparators. Botanical products were well-tolerated across studies, with no serious AEs reported. Despite the limitations such as small sample size, short duration of follow up and varied botanical products, this work still represents the best level of evidence currently available on topical use of botanical products on melasma. Moreover, it should be noted that more well-designed studies are needed before recommending topical botanical products as a viable treatment option for melasma.

Systematic Review Registration

PROSPERO, identifier: CRD42021256328.