bluuebunny/crossref_metadata_2025 huggingface dataset:
Exosomes From miR-19b-3p-Modified ADSCs Inhibit Ferroptosis in Intracerebral Hemorrhage Mice
Authors: Yi Xia , Tang Xiangqi
Published: July 2021

Objectives: Effective treatments for intracerebral hemorrhage (ICH) are limited until now. Ferroptosis, a novel form of iron-dependent cell death, is implicated in neurodegeneration diseases. Here, we attempted to investigate the impact of exosomes from miR-19b-3p-modified adipose-derived stem cells (ADSCs) on ferroptosis in ICH.

Methods: Collagenase was used to induce a mouse model of ICH and hemin was used to induce ferroptosis in cultured neurons. Exosomes were isolated from mimic NC- or miR-19b-3p mimic-transfected ADSCs (ADSCs-MNC-Exos or ADSCs-19bM-Exos, respectively) and then administered to ICH mice or hemin-treated neurons. ICH damage was evaluated by assessing the neurological function of ICH mice and cell viability of neurons. Ferroptosis was evaluated in mouse brains or cultured neurons. The interaction between miR-19b-3p and iron regulatory protein 2 (IRP2) 3′-UTR was analyzed by performing luciferase reporter assay.

Results: Ferroptosis occurred in ICH mice, which also exhibited decreased miR-19b-3p and increased IRP2 expression. IRP2 was a direct target of miR-19b-3p, and IRP2 expression was repressed by ADSCs-19bM-Exos. Importantly, ADSCs-19bM-Exos effectively attenuated hemin-induced cell injury and ferroptosis. Moreover, ADSCs-19bM-Exos administration significantly improved neurologic function and inhibited ferroptosis in ICH mice.

Conclusion: Exosomes from miR-19b-3p-modified ADSCs inhibit ferroptosis in ICH mice.
Accessing Mitochondrial Protein Import in Living Cells by Protein Microinjection
Authors: Bogorodskiy Andrey , Okhrimenko Ivan , Maslov Ivan , Maliar Nina , Burkatovskii Dmitrii , von Ameln Florian , Schulga Alexey , Jakobs Philipp , Altschmied Joachim , Haendeler Judith , Katranidis Alexandros , Sorokin Ivan , Mishin Alexey , Gordeliy Valentin , Büldt Georg , Voos Wolfgang , Gensch Thomas , Borshchevskiy Valentin
Published: July 2021

Mitochondrial protein biogenesis relies almost exclusively on the expression of nuclear-encoded polypeptides. The current model postulates that most of these proteins have to be delivered to their final mitochondrial destination after their synthesis in the cytoplasm. However, the knowledge of this process remains limited due to the absence of proper experimental real-time approaches to study mitochondria in their native cellular environment. We developed a gentle microinjection procedure for fluorescent reporter proteins allowing a direct non-invasive study of protein transport in living cells. As a proof of principle, we visualized potential-dependent protein import into mitochondria inside intact cells in real-time. We validated that our approach does not distort mitochondrial morphology and preserves the endogenous expression system as well as mitochondrial protein translocation machinery. We observed that a release of nascent polypeptides chains from actively translating cellular ribosomes by puromycin strongly increased the import rate of the microinjected pre-protein. This suggests that a substantial amount of mitochondrial translocase complexes was involved in co-translational protein import of endogenously expressed pre-proteins. Our protein microinjection method opens new possibilities to study the role of mitochondrial protein import in cell models of various pathological conditions as well as aging processes.
C1GALT1, Negatively Regulated by miR-181d-5p, Promotes Tumor Progression via Upregulating RAC1 in Lung Adenocarcinoma
Authors: Dong Xiaoxia , Liu Yongyu , Deng Xinzhou , Shao Jun , Tian Shuangyue , Chen Shuang , Huang Rongxin , Lin Ziao , Chen Chunli , Shen Li
Published: July 2021

Glycosyltransferases are frequently dysregulated in lung cancer. Core 1 β 1, 3-galactosyltransferase 1 (C1GALT1), an enzyme highly expressed in various cancers, is correlated with tumor initiation and development. However, the role of C1GALT1 in lung cancer remains poorly understood. In this study, through bioinformatic analysis and clinical validation, we first discovered that C1GALT1 expression was upregulated in lung adenocarcinoma (LUAD) tissues and was closely related to poor prognosis in patients with LUAD. Gain- and loss-of-function experiments showed that C1GALT1 promoted LUAD cell proliferation, migration, and invasion in vitro, as well as tumor formation in vivo. Further investigation demonstrated that RAC1 expression was positively regulated by C1GALT1 in LUAD, whereas silencing Rac1 could reverse C1GALT1-induced tumor growth and metastasis. Moreover, miR-181d-5p was identified as a negative regulator for C1GALT1 in LUAD. As expected, the inhibitory effects of miR-181d-5p on LUAD cell proliferation, migration, and invasion were counteracted by restoration of C1GALT1. In summary, our results highlight the importance of the miR-181d-5p/C1GALT1/RAC1 regulatory axis during LUAD progression. Thus, C1GALT1 may serve as a potential therapeutic target for LUAD.
Cavity-Containing [Fe2L3]4+ Helicates: An Examination of Host-Guest Chemistry and Cytotoxicity
Authors: Lisboa Lynn S. , Riisom Mie , Vasdev Roan A. S. , Jamieson Stephen M. F. , Wright L. James , Hartinger Christian G. , Crowley James D.
Published: July 2021

Two new di(2,2′-bipyridine) ligands, 2,6-bis([2,2′-bipyridin]-5-ylethynyl)pyridine (L1) and bis(4-([2,2′-bipyridin]-5-ylethynyl)phenyl)methane (L2) were synthesized and used to generate two metallosupramolecular [Fe2(L)3](BF4)4 cylinders. The ligands and cylinders were characterized using elemental analysis, electrospray ionization mass spectrometry, UV-vis, 1H-, 13C and DOSY nuclear magnetic resonance (NMR) spectroscopies. The molecular structures of the [Fe2(L)3](BF4)4 cylinders were confirmed using X-ray crystallography. Both the [Fe2(L1)3](BF4)4 and [Fe2(L2)3](BF4)4 complexes crystallized as racemic (rac) mixtures of the ΔΔ (P) and ΛΛ (M) helicates. However, 1H NMR spectra showed that in solution the larger [Fe2(L2)3](BF4)4 was a mixture of the rac-ΔΔ/ΛΛ and meso-ΔΛ isomers. The host-guest chemistry of the helicates, which both feature a central cavity, was examined with several small drug molecules. However, none of the potential guests were found to bind within the helicates. In vitro cytotoxicity assays demonstrated that both helicates were active against four cancer cell lines. The smaller [Fe2(L1)3](BF4)4 system displayed low μM activity against the HCT116 (IC50 = 7.1 ± 0.5 μM) and NCI-H460 (IC50 = 4.9 ± 0.4 μM) cancer cells. While the antiproliferative effects against all the cell lines examined were less than the well-known anticancer drug cisplatin, their modes of action would be expected to be very different.
Solvent-Driven Chirality Switching of a Pillar[4]arene[1]quinone Having a Chiral Amine-Substituted Quinone Subunit
Authors: Liu Chunhong , Yu Zhipeng , Yao Jiabin , Ji Jiecheng , Zhao Ting , Wu Wanhua , Yang Cheng
Published: July 2021

Several new chiral pillar[4]arene[1]quinone derivatives were synthesized by reacting pillar[4]arene[1]quinone (EtP4Q1), containing four 1,4-diethoxybenzene units and one benzoquinone unit, with various chiral amines via Michael addition. Due to the direct introduction of chiral substituents on the rim of pillar[n]arene and the close location of the chiral center to the rim of EtP4Q1, the newly prepared compounds showed unique chiroptical properties without complicated chiral resolution processes, and unprecedented high anisotropy factor of up to −0.018 at the charge transfer absorption band was observed. Intriguingly, the benzene sidearm attached pillar[4]arene[1]quinone derivative 1a showed solvent- and complexation-driven chirality inversion. This work provides a promising potential for absolute asymmetric synthesis of pillararene-based derivatives.
Recovery of Infectious Human Norovirus GII.4 Sydney From Fomites via Replication in Human Intestinal Enteroids
Authors: Overbey Katie N. , Zachos Nicholas C. , Coulter Caroline , Jacangelo Joseph , Schwab Kellogg J.
Published: July 2021

Contamination of fomites by human norovirus (HuNoV) can initiate and prolong outbreaks. Fomite swabbing is necessary to predict HuNoV exposure and target interventions. Historically, swab recovered HuNoV has been measured by molecular methods that detect viral RNA but not infectious HuNoV. The recent development of HuNoV cultivation in human intestinal enteroids (HIEs) enables detection of infectious HuNoV. It is unknown if the swabbing process and swab matrix will allow for cultivation of fomite recovered HuNoV. We used HIEs to culture swab-recovered HuNoV GII.4 Sydney from experimentally infected surfaces—a hospital bed tray (N = 32), door handle (N = 10), and sanitizer dispenser (N = 11). Each surface was swabbed with macrofoam swabs premoistened in PBS plus 0.02% Tween80. Swab eluate was tested for infectious HuNoV by cultivation in HIE monolayers. Infectious HuNoV can be recovered from surfaces inoculated with at least 105 HuNoV genome equivalents/3 cm2. In total, 57% (N = 53) of recovered swabs contained infectious HuNoV detected by HIEs. No difference in percent positive swabs was observed between the three surfaces at p = 0.2. We demonstrate that fomite swabbing can be combined with the HIE method to cultivate high titer infectious HuNoV from the environment, filling a significant gap in HuNoV detection. Currently, high titers of HuNoV are required to measure growth in HIEs and the HIE system precludes absolute quantification of infectious viruses. However, the HIE system can provide a binary indication of infectious HuNoV which enhances existing detection methods. Identification of infectious HuNoVs from swabs can increase monitoring accuracy, enhance risk estimates, and help prevent outbreaks.
Clinical Translation of Three-Dimensional Scar, Diffusion Tensor Imaging, Four-Dimensional Flow, and Quantitative Perfusion in Cardiac MRI: A Comprehensive Review
Authors: Paddock Sophie , Tsampasian Vasiliki , Assadi Hosamadin , Mota Bruno Calife , Swift Andrew J. , Chowdhary Amrit , Swoboda Peter , Levelt Eylem , Sammut Eva , Dastidar Amardeep , Broncano Cabrero Jordi , Del Val Javier Royuela , Malcolm Paul , Sun Julia , Ryding Alisdair , Sawh Chris , Greenwood Richard , Hewson David , Vassiliou Vassilios , Garg Pankaj
Published: July 2021

Cardiovascular magnetic resonance (CMR) imaging is a versatile tool that has established itself as the reference method for functional assessment and tissue characterisation. CMR helps to diagnose, monitor disease course and sub-phenotype disease states. Several emerging CMR methods have the potential to offer a personalised medicine approach to treatment. CMR tissue characterisation is used to assess myocardial oedema, inflammation or thrombus in various disease conditions. CMR derived scar maps have the potential to inform ablation therapy—both in atrial and ventricular arrhythmias. Quantitative CMR is pushing boundaries with motion corrections in tissue characterisation and first-pass perfusion. Advanced tissue characterisation by imaging the myocardial fibre orientation using diffusion tensor imaging (DTI), has also demonstrated novel insights in patients with cardiomyopathies. Enhanced flow assessment using four-dimensional flow (4D flow) CMR, where time is the fourth dimension, allows quantification of transvalvular flow to a high degree of accuracy for all four-valves within the same cardiac cycle. This review discusses these emerging methods and others in detail and gives the reader a foresight of how CMR will evolve into a powerful clinical tool in offering a precision medicine approach to treatment, diagnosis, and detection of disease.
The Expression Patterns and Roles of Lysyl Oxidases in Aortic Dissection
Authors: Yi Xin , Zhou Yi , Chen Yue , Feng Xin , Liu Chang , Jiang Ding-Sheng , Geng Jing , Li Xiaoyan , Jiang Xuejun , Fang Ze-Min
Published: July 2021

Background: Lysyl oxidases (LOXs), including LOX, LOXL1, LOXL2, LOXL3, and LOXL4, catalyze the formation of a cross-link between elastin (ELN) and collagen. Multiple LOX mutations have been shown to be associated with the occurrence of aortic dissection (AD) in humans, and LOX-knockout mice died during the perinatal period due to aortic aneurysm and rupture. However, the expression levels and roles of other LOX members in AD remain unknown.

Methods: A total of 33 aorta samples of AD and 15 normal aorta were collected for LOXs mRNA and protein levels detection. We also analyzed the datasets of AD in GEO database through bioinformatics methods. LOXL2 and LOXL3 were knocked down in primary cultured human aortic smooth muscle cells (HASMCs) via lentivirus.

Results: Here, we show that the protein levels of LOXL2 and LOXL3 are upregulated, while LOXL4 is downregulated in AD subjects compared with non-AD subjects, but comparable protein levels of LOX and LOXL1 are detected. Knockdown of LOXL2 suppressed MMP2 expression, the phosphorylation of AKT (p-AKT) and S6 (p-S6), but increased the mono-, di-, tri-methylation of H3K4 (H3K4me1/2/3), H3K9me3, and p-P38 levels in HASMCs. These results indicate that LOXL2 is involved in regulation of the extracellular matrix (ECM) in HASMCs. In contrast, LOXL3 knockdown inhibited PCNA and cyclin D1, suppressing HASMC proliferation. Our results suggest that in addition to LOX, LOXL2 and LOXL3 are involved in the pathological process of AD by regulating ECM and the proliferation of HASMCs, respectively. Furthermore, we found that LOXL2 and LOXL4 was inhibited by metformin and losartan in HASMCs, which indicated that LOXL2 and LOXL4 are the potential targets that involved in the therapeutic effects of metformin and losartan on aortic or aneurysm expansion.

Conclusions: Thus, differential regulation of LOXs might be a novel strategy to prevent or treat AD.
Orbital and Millennial Variations in Sea Ice in the Southwestern Okhotsk Sea Since the Last Interglacial Period and Their Implications
Authors: Wang Anqi , Yao Zhengquan , Shi Xuefa , Wang Kunshan , Zou Jianjun , Liu Yanguang , Wu Yonghua , Gorbarenko Sergey A.
Published: July 2021

Sea ice in the Okhotsk Sea plays a significant role in global climate change. However, the history and mechanism of changes in sea ice spanning the last glacial cycle remain controversial. In this study, an 8.8 m core (LV55-40-1) was recovered from the southwestern Okhotsk Sea that contains a continuous sea ice record over the past ∼110 kyr. The sand fraction and dropstones were used as ice-rafted debris proxies to reconstruct the history of sea ice variations over the last ∼110 kyr and to determine the underlying causes on orbital and millennial timescales. Sea ice expansions occurred during MIS 5b, MIS 4, mid-MIS 3, and early MIS 1, which were controlled mainly by decreased autumn insolation on an orbital timescale. Superimposed on the orbital-scale changes, millennial-scale variations in sea ice were also observed, with 19 expansion events that coincided with cold Dansgaard-Oeschger stadials. Millennial scale sea ice variations were most likely controlled by both the Arctic oscillation and the East Asian summer monsoon. During periods of negative Arctic oscillation patterns, decreased air temperatures over the Okhotsk Sea caused more active sea ice formation. Such conditions could have been reinforced, by a reduced influence of warm advection at the surface of the Okhotsk Sea caused by decreased discharge from the Amur River that resulted from a weakened East Asian summer monsoon during cold stadials.
Glyphosate Herbicide: Reproductive Outcomes and Multigenerational Effects
Authors: Milesi María Mercedes , Lorenz Virginia , Durando Milena , Rossetti María Florencia , Varayoud Jorgelina
Published: July 2021

Glyphosate base herbicides (GBHs) are the most widely applied pesticides in the world and are mainly used in association with GBH-tolerant crop varieties. Indiscriminate and negligent use of GBHs has promoted the emergence of glyphosate resistant weeds, and consequently the rise in the use of these herbicides. Glyphosate, the active ingredient of all GBHs, is combined with other chemicals known as co-formulants that enhance the herbicide action. Nowadays, the safety of glyphosate and its formulations remain to be a controversial issue, as evidence is not conclusive whether the adverse effects are caused by GBH or glyphosate, and little is known about the contribution of co-formulants to the toxicity of herbicides. Currently, alarmingly increased levels of glyphosate have been detected in different environmental matrixes and in foodstuff, becoming an issue of social concern. Some in vitro and in vivo studies have shown that glyphosate and its formulations exhibit estrogen-like properties, and growing evidence has indicated they may disrupt normal endocrine function, with adverse consequences for reproductive health. Moreover, multigenerational effects have been reported and epigenetic mechanisms have been proved to be involved in the alterations induced by the herbicide. In this review, we provide an overview of: i) the routes and levels of human exposure to GBHs, ii) the potential estrogenic effects of glyphosate and GBHs in cell culture and animal models, iii) their long-term effects on female fertility and mechanisms of action, and iv) the consequences on health of successive generations.
Polyaneuploid Cancer Cell Dormancy: Lessons From Evolutionary Phyla
Authors: Kostecka Laurie G. , Pienta Kenneth J. , Amend Sarah R.
Published: July 2021

Dormancy is a key survival strategy in many organisms across the tree of life. Organisms that utilize some type of dormancy (hibernation, aestivation, brumation, diapause, and quiescence) are able to survive in habitats that would otherwise be uninhabitable. Induction into dormant states is typically caused by environmental stress. While organisms are dormant, their physical activity is minimal, and their metabolic rates are severely depressed (hypometabolism). These metabolic reductions allow for the conservation and distribution of energy while conditions in the environment are poor. When conditions are more favorable, the organisms are then able to come out of dormancy and reengage in their environment. Polyaneuploid cancer cells (PACCs), proposed mediators of cancer metastasis and resistance, access evolutionary programs and employ dormancy as a survival mechanism in response to stress. Quiescence, the type of dormancy observed in PACCs, allows these cells the ability to survive stressful conditions (e.g., hypoxia in the microenvironment, transiting the bloodstream during metastasis, and exposure to chemotherapy) by downregulating and altering metabolic function, but then increasing metabolic activities again once stress has passed. We can gain insights regarding the mechanisms underlying PACC dormancy by looking to the evolution of dormancy in different organisms.
Watch Out for a Second SNP: Focus on Multi-Nucleotide Variants in Coding Regions and Rescued Stop-Gained
Authors: Degalez Fabien , Jehl Frédéric , Muret Kévin , Bernard Maria , Lecerf Frédéric , Lagoutte Laetitia , Désert Colette , Pitel Frédérique , Klopp Christophe , Lagarrigue Sandrine
Published: July 2021

Most single-nucleotide polymorphisms (SNPs) are located in non-coding regions, but the fraction usually studied is harbored in protein-coding regions because potential impacts on proteins are relatively easy to predict by popular tools such as the Variant Effect Predictor. These tools annotate variants independently without considering the potential effect of grouped or haplotypic variations, often called “multi-nucleotide variants” (MNVs). Here, we used a large RNA-seq dataset to survey MNVs, comprising 382 chicken samples originating from 11 populations analyzed in the companion paper in which 9.5M SNPs— including 3.3M SNPs with reliable genotypes—were detected. We focused our study on in-codon MNVs and evaluate their potential mis-annotation. Using GATK HaplotypeCaller read-based phasing results, we identified 2,965 MNVs observed in at least five individuals located in 1,792 genes. We found 41.1% of them showing a novel impact when compared to the effect of their constituent SNPs analyzed separately. The biggest impact variation flux concerns the originally annotated stop-gained consequences, for which around 95% were rescued; this flux is followed by the missense consequences for which 37% were reannotated with a different amino acid. We then present in more depth the rescued stop-gained MNVs and give an illustration in theSLC27A4gene. As previously shown in human datasets, our results in chicken demonstrate the value of haplotype-aware variant annotation, and the interest to consider MNVs in the coding region, particularly when searching for severe functional consequence such as stop-gained variants.
GGC Repeat Expansion in the NOTCH2NLC Gene Is Associated With a Phenotype of Predominant Motor–Sensory and Autonomic Neuropathy
Authors: Wang Hui , Yu Jiaxi , Yu Meng , Deng Jianwen , Zhang Wei , Lv He , Liu Jing , Shi Xin , Liang Wei , Jia Zhirong , Hong Daojun , Meng Lingchao , Wang Zhaoxia , Yuan Yun
Published: July 2021

There is still a considerable proportion of patients with inherited peripheral neuropathy (IPN) whose pathogenic genes are unknown. This study was intended to investigate whether the GGC repeat expansion in the NOTCH2NLC is presented in some patients with IPN. A total of 142 unrelated mainland Chinese patients with highly suspected diagnosis of IPN without any known causative gene were recruited. Repeat-primed polymerase chain reaction (RP-PCR) was performed to screen GGC repeat expansion in NOTCH2NLC, followed by fluorescence amplicon length analysis-PCR (AL-PCR) to determine the GGC repeat size. Detailed clinical data as well as nerve, muscle, and skin biopsy were reviewed and analyzed in the NOTCH2NLC-related IPN patients. In total, five of the 142 patients (3.52%) were found to have pathogenic GGC expansion in NOTCH2NLC, with repeat size ranging from 126 to 206 repeats. All the NOTCH2NLC-related IPN patients presented with adult-onset motor–sensory and autonomic neuropathy that predominantly affected the motor component of peripheral nerves. While tremor and irritating dry cough were noted in four-fifths of the patients, no other signs of the central nervous system were presented. Electrophysiological studies revealed both demyelinating and axonal changes of polyneuropathy that were more severe in lower limbs and asymmetrically in upper limbs. Sural nerve pathology was characterized by multiple fibers with thin myelination, indicating a predominant demyelinating process. Muscle pathology was consistent with neuropathic changes. P62-positive intranuclear inclusions were observed in nerve, skin, and muscle tissues. Our study has demonstrated that GGC expansion in NOTCH2NLC is associated with IPN presenting as predominant motor–sensory and autonomic neuropathy, which expands the phenotype of the NOTCH2NLC-related repeat expansion spectrum. Screening of GGC repeat expansions in the NOTCH2NLC should be considered in patients presenting with peripheral neuropathy with tremor and irritating dry cough.
Critical Reflexivity in Political Ecology Research: How can the Covid-19 Pandemic Transform us Into Better Researchers?
Authors: Gonda Noémi , Leder Stephanie , González-Hidalgo Marien , Chiwona-Karltun Linley , Stiernström Arvid , Hajdu Flora , Fischer Klara , Asztalos Morell Ildikó , Kadfak Alin , Arvidsson Anna
Published: July 2021

It is not just the world but our ways of producing knowledge that are in crisis. The Covid-19 pandemic has exposed our interconnected vulnerabilities in ways never seen before while underscoring the need for emancipation in particular from the hegemonic knowledge politics that underpin “business-as-usual” academic research that have both contributed to and failed to address the systemic challenges laid bare by the pandemic. Political ecologists tasked with knowledge generation on vulnerabilities and their underlying power processes are particularly well placed to envision such emancipatory processes. While pausing physically due to travel restrictions, as researchers in political ecology and rural development at the same university department, we want to make a stop to radically rethink our intellectual engagements. In this article, we aim to uncover “sanitized” aspects of research encounters, and theorize on the basis of anecdotes, feelings and informal discussions—“data” that is often left behind in fieldwork notes and personal diaries of researchers—, the ways in which our own research practices hamper or can be conducive to emancipation in times of multiple interconnected health, political, social, and environmental crises. We do so through affective autoethnography and resonances on our research encounters during the pandemic: with people living in Swedish Sapmi, with African students in our own “Global North” university department and with research partners in Nepal. We use a threefold focus on interconnectedness, uncertainty and challenging hegemonic knowledge politics as our analytical framework. We argue that acknowledging the roles of emotions and affect can 1) help embraceinterconnectednessin research encounters; 2) enable us to work withuncertaintyrather than “hard facts” in knowledge production processes; and 3) contribute to challenginghegemonic knowledge production. Opening up for emotions in research helps us to embrace the relational character of vulnerability as a pathway to democratizing power relations and to move away from its oppressive and colonial modes still present in universities and research centers. Our aim is to contribute to envisioning post-Covid-19 political ecology and rural development research that is critically reflexive and that contributes to the emergence of a new ethics of producing knowledge.
Protective Role of Kynurenine 3-Monooxygenase in Allograft Rejection and Tubular Injury in Kidney Transplantation
Authors: Lassiter Randi , Merchen Todd D. , Fang Xuexiu , Wang Youli
Published: July 2021

Renal tubular epithelial cells (TECs) are the primary targets of ischemia–reperfusion injury (IRI) and rejection by the recipient’s immune response in kidney transplantation (KTx). However, the molecular mechanism of rejection and IRI remains to be identified. Our previous study demonstrated that kynurenine 3-monooxygenase (KMO) and kynureninase were reduced in ischemia–reperfusion procedure and further decreased in rejection allografts among mismatched pig KTx. Herein, we reveal that TEC injury in acutely rejection allografts is associated with alterations of Bcl2 family proteins, reduction of tight junction protein 1 (TJP1), and TEC-specific KMO. Three cytokines, IFNγ, TNFα, and IL1β, reported in our previous investigation were identified as triggers of TEC injury by altering the expression of Bcl2, BID, and TJP1. Allograft rejection and TEC injury were always associated with a dramatic reduction of KMO. 3HK and 3HAA, as direct and downstream products of KMO, effectively protected TEC from injury via increasing expression of Bcl-xL and TJP1. Both 3HK and 3HAA further prevented allograft rejection by inhibiting T cell proliferation and up-regulating aryl hydrocarbon receptor expression. Pig KTx with the administration of DNA nanoparticles (DNP) that induce expression of indoleamine 2,3-dioxygenase (IDO) and KMO to increase 3HK/3HAA showed an improvement of allograft rejection as well as murine skin transplant in IDO knockout mice with the injection of 3HK indicated a dramatic reduction of allograft rejection. Taken together, our data provide strong evidence that reduction of KMO in the graft is a key mediator of allograft rejection and loss. KMO can effectively improve allograft outcome by attenuating allograft rejection and maintaining graft barrier function.
Dectin-1 Controls TSLP-Induced Th2 Response by Regulating STAT3, STAT6, and p50-RelB Activities in Dendritic Cells
Authors: Gu Chao , Upchurch Katherine , Horton Joshua , Wiest Mathew , Zurawski Sandra , Millard Mark , Kane Robert R. , Joo HyeMee , Miller Lisa A. , Oh SangKon
Published: July 2021

The epithelium-associated cytokine thymic stromal lymphopoietin (TSLP) can induce OX40L and CCL17 expression by myeloid dendritic cells (mDCs), which contributes to aberrant Th2-type immune responses. Herein, we report that such TSLP-induced Th2-type immune response can be effectively controlled by Dectin-1, a C-type lectin receptor expressed by mDCs. Dectin-1 stimulation induced STAT3 activation and decreased the transcriptional activity of p50-RelB, both of which resulted in reduced OX40L expression on TSLP-activated mDCs. Dectin-1 stimulation also suppressed TSLP-induced STAT6 activation, resulting in decreased expression of the Th2 chemoattractant CCL17. We further demonstrated that Dectin-1 activation was capable of suppressing ragweed allergen (Amb a 1)-specific Th2-type T cell response in allergy patientsex vivoand house dust mite allergen (Der p 1)-specific IgE response in non-human primatesin vivo. Collectively, this study provides a molecular explanation of Dectin-1-mediated suppression of Th2-type inflammatory responses and suggests Dectin-1 as a target for controlling Th2-type inflammation.
BHLHE40 Regulates IL-10 and IFN-γ Production in T Cells but Does Not Interfere With Human Type 1 Regulatory T Cell Differentiation
Authors: Uyeda Molly Javier , Freeborn Robert A. , Cieniewicz Brandon , Romano Rosa , Chen Ping (Pauline) , Liu Jeffrey Mao-Hwa , Thomas Benjamin , Lee Esmond , Cepika Alma-Martina , Bacchetta Rosa , Roncarolo Maria Grazia
Published: July 2021

Type 1 regulatory T (Tr1) cells are subset of peripherally induced antigen-specific regulatory T cells. IL-10 signaling has been shown to be indispensable for polarization and function of Tr1 cells. However, the transcriptional machinery underlying human Tr1 cell differentiation and function is not yet elucidated. To this end, we performed RNA sequencing on ex vivo human CD49b+LAG3+ Tr1 cells. We identified the transcription factor, BHLHE40, to be highly expressed in Tr1 cells. Even though Tr1 cells characteristically produce high levels of IL-10, we found that BHLHE40 represses IL-10 and increases IFN-γ secretion in naïve CD4+ T cells. Through CRISPR/Cas9-mediated knockout, we determined that IL10 significantly increased in the sgBHLHE40-edited cells and BHLHE40 is dispensable for naïve CD4+ T cells to differentiate into Tr1 cells in vitro. Interestingly, BHLHE40 overexpression induces the surface expression of CD49b and LAG3, co-expressed surface molecules attributed to Tr1 cells, but promotes IFN-γ production. Our findings uncover a novel mechanism whereby BHLHE40 acts as a regulator of IL-10 and IFN-γ in human CD4+ T cells.
Elevated Biomarkers of NETosis in the Serum of Pediatric Patients With Type 1 Diabetes and Their First-Degree Relatives
Authors: Klocperk Adam , Vcelakova Jana , Vrabcova Petra , Zentsova Irena , Petruzelkova Lenka , Sumnik Zdenek , Pruhova Stepanka , Sediva Anna , Parackova Zuzana
Published: July 2021

Type 1 diabetes (T1D) is an autoimmune disorder with unambiguous involvement of both innate and adaptive immune mechanisms in the destruction of pancreatic beta cells. Recent evidence demonstrated that neutrophils infiltrate the pancreas prior to disease onset and therein extrude neutrophil extracellular traps (NETs), web-like structures of DNA and nuclear proteins with a strong pro-inflammatory biologic activity. Our previous work showed that T1D NETs activate dendritic cells, which consequently induce IFNγ-producing Th1 lymphocytes. The aim of this study was to assess direct ex vivo biomarkers of NETosis in the serum of recent onset and long-term pediatric T1D patients, their first-degree relatives and healthy controls. To this end we evaluated serum levels of myeloperoxidase (MPO), neutrophil elastase (NE), proteinase 3 (PR3), protein arginine deiminase 4 (PAD4), LL37 and cell-free DNA-histone complexes in sex- and age-matched cohorts of T1D first-degree relatives, recent-onset T1D patients, and in patients 12 months after clinical manifestation of the disease. Our data shows that disease onset is accompanied by peripheral neutrophilia and significant elevation of MPO, NE, PR3, PAD4 and cell-free DNA-histone complexes. Most biomarkers subsequently decrease but do not always normalize in long-term patients. First-degree relatives displayed an intermediate phenotype, except for remarkably high levels of LL37. Together, this report provides evidence for the presence of ongoing NETosis in pediatric patients with T1D at time of clinical manifestation of the disease, which partly subsides in subsequent years.
Staphylococcus aureus Vaccine Research and Development: The Past, Present and Future, Including Novel Therapeutic Strategies
Authors: Clegg Jonah , Soldaini Elisabetta , McLoughlin Rachel M. , Rittenhouse Stephen , Bagnoli Fabio , Phogat Sanjay
Published: July 2021

Staphylococcus aureusis one of the most important human pathogens worldwide. Its high antibiotic resistance profile reinforces the need for new interventions like vaccines in addition to new antibiotics. Vaccine development efforts againstS. aureushave failed so far however, the findings from these human clinical and non-clinical studies provide potential insight for such failures. Currently, research is focusing on identifying novel vaccine formulations able to elicit potent humoral and cellular immune responses. Translational science studies are attempting to discover correlates of protection using animal models as well asin vitroandex vivomodels assessing efficacy of vaccine candidates. Several new vaccine candidates are being tested in human clinical trials in a variety of target populations. In addition to vaccines, bacteriophages, monoclonal antibodies, centyrins and new classes of antibiotics are being developed. Some of these have been tested in humans with encouraging results. The complexity of the diseases and the range of the target populations affected by this pathogen will require a multipronged approach using different interventions, which will be discussed in this review.
The TNFR1 Antagonist Atrosimab Is Therapeutic in Mouse Models of Acute and Chronic Inflammation
Authors: Richter Fabian , Williams Sarah K. , John Katharina , Huber Carina , Vaslin Camille , Zanker Henri , Fairless Richard , Pichi Kira , Marhenke Silke , Vogel Arndt , Dhaen Marie-Ann , Herrmann Stefanie , Herrmann Andreas , Pfizenmaier Klaus , Bantel Heike , Diem Ricarda , Kontermann Roland E. , Fischer Roman
Published: July 2021

Therapeutics that block tumor necrosis factor (TNF), and thus activation of TNF receptor 1 (TNFR1) and TNFR2, are clinically used to treat inflammatory diseases such as rheumatoid arthritis, inflammatory bowel disease and psoriasis. However, TNFR1 and TNFR2 work antithetically to balance immune responses involved in inflammatory diseases. In particular, TNFR1 promotes inflammation and tissue degeneration, whereas TNFR2 contributes to immune modulation and tissue regeneration. We, therefore, have developed the monovalent antagonistic anti-TNFR1 antibody derivative Atrosimab to selectively block TNFR1 signaling, while leaving TNFR2 signaling unaffected. Here, we describe that Atrosimab is highly stable at different storage temperatures and demonstrate its therapeutic efficacy in mouse models of acute and chronic inflammation, including experimental arthritis, non-alcoholic steatohepatitis (NASH) and experimental autoimmune encephalomyelitis (EAE). Our data support the hypothesis that it is sufficient to block TNFR1 signaling, while leaving immune modulatory and regenerative responses via TNFR2 intact, to induce therapeutic effects. Collectively, we demonstrate the therapeutic potential of the human TNFR1 antagonist Atrosimab for treatment of chronic inflammatory diseases.
SARS-CoV-2 Accessory Proteins in Viral Pathogenesis: Knowns and Unknowns
Authors: Redondo Natalia , Zaldívar-López Sara , Garrido Juan J. , Montoya Maria
Published: July 2021

There are still many unanswered questions concerning viral SARS-CoV-2 pathogenesis in COVID-19. Accessory proteins in SARS-CoV-2 consist of eleven viral proteins whose roles during infection are still not completely understood. Here, a review on the current knowledge of SARS-CoV-2 accessory proteins is summarized updating new research that could be critical in understanding SARS-CoV-2 interaction with the host. Some accessory proteins such as ORF3b, ORF6, ORF7a and ORF8 have been shown to be important IFN-I antagonists inducing an impairment in the host immune response. In addition, ORF3a is involved in apoptosis whereas others like ORF9b and ORF9c interact with cellular organelles leading to suppression of the antiviral response in infected cells. However, possible roles of ORF7b and ORF10 are still awaiting to be described. Also, ORF3d has been reassigned. Relevant information on the knowns and the unknowns in these proteins is analyzed, which could be crucial for further understanding of SARS-CoV-2 pathogenesis and to design strategies counteracting their actions evading immune responses in COVID-19.
Freshwater, Landlocked Grand Lake Strain of Atlantic Salmon (Salmo salar L.) as a Potential Genetic Source of Long Chain Polyunsaturated Fatty Acids Synthesis
Authors: Colombo Stefanie M. , Emam Mohamed , Peterson Brian C. , Hall Jennifer R. , Burr Gary , Zhang Zeyu , Rise Matthew L.
Published: July 2021

Selection efforts focused on adaptation to plant-based diets, particularly the ability to synthesize polyunsaturated fatty acids (PUFA), are now emerging in aquaculture. Landlocked salmon (Grand Lake population; GL) may differ from the commercial Saint John River (SJR) strain in terms of PUFA metabolism. The objective of this study was to determine if GL salmon can contribute toward broodstock selection for enhanced PUFA synthesis. Two diets containing either fish oil (FO) or plant-based oil (FO-free) were fed to the SJR and GL strains (∼58 g/fish) for 16 weeks. Growth, liver, and muscle fatty acid (FA) content, and transcript expression of lipid metabolism and inflammation-related genes were evaluated. GL salmon fed the FO diet showed reduced growth compared to SJR salmon (fed either diet); however, GL salmon fed the FO-free diet, growth was not significantly different compared to any group. In liver, SJR salmon fed the FO-free diet had higher levels of n-6 PUFAs (21.9%) compared to GL fed the same diet (15.9%); while GL salmon fed the FO-free diet had higher levels of monounsaturated FAs (48.9%) compared with SJR salmon fed the same diet (35.7%). 20:5n-3 and 22:6n-3 were the same in GL and SJR salmon liver and muscle, respectively, fed the FO-free diet. In liver, GL salmon fed the FO-free diet had higher acac and acly compared to all treatments and had higher fasb compared to both strains fed the FO-diet. GL salmon fed the FO-free diet had higher cd36c and fabp3b in liver compared to GL salmon fed the FO diet and SJR salmon fed either diet. GL salmon fed the FO-free diet had higher lect2a and pgds in liver compared to SJR salmon fed the FO-free diet. In muscle, GL salmon fed the FO-free diet had higher fadsd5 and fadsd6b compared with both strains fed the FO diet. These results suggest there is a genetic basis behind the potential for GL salmon to utilize FO-free diets more efficiently than SJR salmon, with regards to FA metabolism.
Monsoon Influence on the Island Mass Effect Around the Maldives and Sri Lanka
Authors: Su Danielle , Wijeratne Sarath , Pattiaratchi Charitha Bandula
Published: July 2021

The monsoon circulation in the Northern Indian Ocean (NIO) is unique since it develops in response to the bi-annual reversing monsoonal winds, with the ocean currents mirroring this change through directionality and intensity. The interaction between the reversing currents and topographic features have implications for the development of the Island Mass Effect (IME) in the NIO. The IME in the NIO is characterized by areas of high chlorophyll concentrations identified through remote sensing to be located around the Maldives and Sri Lanka in the NIO. The IME around the Maldives was observed to reverse between the monsoons to downstream of the incoming monsoonal current whilst a recirculation feature known as the Sri Lanka Dome (SLD) developed off the east coast of Sri Lanka during the Southwest Monsoon (SWM). To understand the physical mechanisms underlying this monsoonal variability of the IME, a numerical model based on the Regional Ocean Modeling System (ROMS) was implemented and validated. The model was able to simulate the regional circulation and was used to investigate the three-dimensional structure of the IME around the Maldives and Sri Lanka in terms of its temperature and velocity. Results revealed that downwelling processes were prevalent along the Maldives for both monsoon periods but was applicable only to latitudes above 4°N since that was the extent of the monsoon current influence. For the Maldives, atolls located south of 4°N, were influenced by the equatorial currents. Around Sri Lanka, upwelling processes were responsible for the IME during the SWM but with strong downwelling during the NEM. In addition, there were also regional differences in intra-seasonal variability for these processes. Overall, the strength of the IME processes was closely tied to the monsoon current intensity and was found to reach its peak when the monsoon currents were at the maximum.
Tidal and Physicochemical Effects on Phytoplankton Community Variability at Tagus Estuary (Portugal)
Authors: Cereja Rui , Brotas Vanda , Cruz Joana P. C. , Rodrigues Marta , Brito Ana C.
Published: July 2021

The Tagus Estuary is one of the largest estuaries in Europe and merges large urban and industrial areas. Understanding phytoplankton community variability is key for an appropriate assessment of the estuarine ecological status. The objective of the present study was to assess the importance of the tidal influence over the phytoplankton community and to evaluate its main drivers of variation. Weekly sampling was performed at two stations on the Tagus Estuary with different anthropogenic pressures (Alcântara and Barreiro). The sampling covered periods with different tidal amplitude. Alcântara presented both the lowest and highest concentrations of dissolved inorganic nitrogen (DIN) and orthophosphate concentration (DIP), depending on the tidal height. Such high variability in this sampling station is probably due to its proximity to a sewage treatment station outfall and to the estuary mouth. In the present study, both seasonal and tidal variations influenced the chlorophyll a concentration of which the tidal cycle explained up to 50% of the chlorophyll a variations. Chlorophyll a displayed a seasonal trend with two peaks of phytoplankton biomass between spring and mid-summer. The main drivers of chlorophyll a variation were radiation, water temperature, tidal amplitude, salinity, river discharge, and the inorganic nutrients DIN and DSi. The estuarine phytoplankton community was mainly dominated by Bacillariophyceae, especially at Alcântara. Bacillariophyceae were less important at Barreiro, where communities had a higher representation from other phytoplankton groups, such as Cryptophyceae and Prasinophyceae. The drivers of variability in the community composition were similar to those influencing the total biomass. In conclusion, the spring-neap tidal cycle strongly influenced the phytoplankton community, both in terms of biomass and community composition. Of the several tidal conditions, spring tides were the tidal condition that presented both higher biomass and higher Bacillariophyceae representativity in the community.
Drivers of Atmosphere-Ocean CO2 Flux in Northern Norwegian Fjords
Authors: Aalto Nerea J. , Campbell Karley , Eilertsen Hans C. , Bernstein Hans C.
Published: July 2021

High-latitude fjords and continental shelves are shown to be sinks for atmospheric CO2, yet large spatial-temporal variability and poor regional coverage of sea-air CO2 flux data, especially from fjord systems, makes it difficult to scale our knowledge on how they contribute to atmospheric carbon regulation. The magnitude and seasonal variability of atmosphere-sea CO2 flux was investigated in high-latitude northern Norwegian coastal areas over 2018 and 2019, including four fjords and one coastal bay. The aim was to assess the physical and biogeochemical factors controlling CO2 flux and partial pressure of CO2 in surface water via correlation to physical oceanographic and biological measurements. The results show that the study region acts as an overall atmospheric CO2 sink throughout the year, largely due to the strong undersaturation of CO2 relative to atmospheric concentrations. Wind speed exerted the strongest influence on the instantaneous rate of sea-air CO2 exchange, while exhibiting high variability. We concluded that the northernmost fjords (Altafjord and Porsangerfjord) showed stronger potential for instantaneous CO2 uptake due to higher wind speeds. We also found that fixation of CO2 was likely a significant factor controlling ΔpCO2 from April to June, which followed phenology of spring phytoplankton blooms at each location. Decreased ΔpCO2 and the resulting sea-air CO2 flux was observed in autumn due to a combined reduction of the mixed layer with entrain of high CO2 subsurface water, damped biological activity and higher surface water temperatures. This study provides the first measurements of atmospheric CO2 flux in these fjord systems and therefore an important new baseline for gaining a better understanding on how the northern Norwegian coast and characteristic fjord systems participate in atmosphere carbon regulation.
Statistical Assessment of Parameters Affecting Firebrand Pile Heat Transfer to Surfaces
Authors: Bearinger Elias , Lattimer Brian Y. , Hodges Jonathan L. , Rippe Christian , Kapahi Anil
Published: July 2021

Firebrands are known to cause ignition of structures far from the primary fire front, resulting in significant damage to structures before firefighting can be attempted. To make structures more resilient to firebrand ignition, a better understanding of the heat transfer from firebrands to surfaces is needed. This paper provides a statistical assessment of different factors expected to have an impact on the heat flux from firebrand piles to a flat surface. The factors included in the study were wood moisture content, wood type (hardwood or softwood), wood density, wood state (live, dead, or artificial), wind speed, pile mass, firebrand diameter, and firebrand length. Using design of experiments, test matrices were developed that permitted a statistical analysis to be performed on the data. This statistical analysis was used to quantify which factors had a statistically significant impact on the heat flux from the pile as well as ranking the importance of the different factors. Artificial firebrands were found to have statistically higher heat fluxes compared with natural firebrands. Other factors that had a statistically significant impact on the heat flux were wind speed, firebrand length, and firebrand length-diameter interaction. Firebrand aspect ratio (related to the firebrand length-diameter interaction) is directly related to the pile porosity, which is a measure of the volume of air in the pile. Increasing the aspect ratio (which increases the pile porosity) results in higher heat fluxes across a larger region of the pile and was found to be an important factor. Firebrand diameter and pile mass were found to affect the burning duration but not as significantly as other parameters. The number of firebrands in the pile was also observed to potentially affect the heat flux, with a critical number required to reach the highest heat flux for a given firebrand geometry.