Risk perception and knowledge of COVID-19 in patients with celiac disease

BACKGROUND We recently demonstrated that the odds of contracting coronavirus disease 2019(COVID-19)in patients with celiac disease(CeD)is similar to that of the general population.However,how patients with CeD perceive their COVID-19 risk may differ from their actual risk.AIM To investigate risk perceptions of contracting COVID-19 in patients with CeD and determine the factors that may influence their perception.METHODS We distributed a survey throughout 10 countries between March and June 2020 and collected data on demographics,diet,COVID-19 testing,and risk perceptions of COVID-19 in patients with CeD.Participants were recruited through various celiac associations,clinic visits,and social media.Risk perception was assessed by asking individuals whether they believe patients with CeD are at an increased risk of contracting COVID-19 when compared to the general population.Logistic regression was used to determine the influencing factors associated with COVID-19 risk perception,such as age,sex,adherence to a gluten-free diet(GFD),and comorbidities such as cardiac conditions,respiratory conditions,and diabetes.Data was presented as adjusted odds ratios(aORs)RESULTS A total of 10737 participants with CeD completed the survey.From them,6019(56.1%)patients with CeD perceived they were at a higher risk or were unsure if they were at a higher risk of contracting COVID-19 compared to the non-CeD population.A greater proportion of patients with CeD perceived an increased risk of contracting COVID-19 when compared to infections in general due to their CeD(56.1%vs 26.7%,P<0.0001).Consequently,34.8%reported taking extra COVID-19 precautions as a result of their CeD.Members of celiac associations were less likely to perceive an increased risk of COVID-19 when compared to non-members(49.5%vs 57.4%,P<0.0001).Older age(aOR:0.99;95%CI:0.99 to 0.99,P<0.001),male sex(aOR:0.84;95%CI:0.76 to 0.93,P=0.001),and strict adherence to a GFD(aOR:0.89;95%CI:0.82 to 0.96,P=0.007)were associated with a lower perception of COVID-19 risk and the presence of comorbidities was associated with a higher perception of COVID-19 risk(aOR:1.38;95%CI:1.22 to 1.54,P<0.001).CONCLUSION Overall,high levels of risk perceptions,such as those found in patients with CeD,may increase an individual’s pandemic-related stress and contribute to negative mental health consequences.Therefore,it is encouraged that public health officials maintain consistent communication with the public and healthcare providers with the celiac community.Future studies specifically evaluating mental health in CeD could help determine the consequences of increased risk perceptions in this population.

How the Bcl-2 family of proteins interact to regulate apoptosis

Commitment of cells to apoptosis is governed largely by protein-protein interactions between members of the Bcl-2 protein family. Its three sub-families have distinct roles： the BH3-only proteins trigger apoptosis by binding via their BH3 domain to pro-survival relatives, while the pro-apoptotic Bax and Bak have an essential downstream role involving disruption of organellar membranes and induction of caspase activation. The BH3-only proteins act as damage sensors, held inert until their activation by stress signals. Once activated, they were thought to bind promiscuously to pro-survival protein targets but unexpected selectivity has recently emerged from analysis of their interactions. Some BH3-only proteins also bind to Bax and Bak. Whether Bax and Bak are activated directly by these BH3-only proteins, or indirectly as a consequence of BH3-only proteins neutralizing their pro-survival targets is the subject of intense debate. Regardless of this, a detailed understanding of the interactions between family members, which are often selective, has notable implications for designing anti-cancer drugs to target the Bcl-2 family.

Colorectal cancer biomarkers: To be or not to be? Cautionary tales from a road well travelled

Colorectal cancer(CRC)is the second most common cause of cancer-related death worldwide and places a major economic burden on the global health care system.The time frame for development from premalignant to malignant disease typically spans 10-15 years,and this latent period provides an ideal opportunity for early detection and intervention to improve patient outcomes.Currently,early diagnosis of CRC is hampered by a lack of suitable non-invasive biomarkers that are clinically or economically acceptable for populationbased screening.New blood-based protein biomarkers for early detection of CRC are therefore urgently required.The success of clinical biomarker discovery and validation studies is critically dependent on understanding and adjusting for potential experimental,analytical,and biological factors that can interfere with the robust interpretation of results.In this review we outline some important considerations for research groups undertaking biomarker research with exemplars from our studies.Implementation of experimental strategies to minimise the potential effects of these problems will facilitate the identification of panels of biomarkers with the sensitivity and specificity required for the development of successful tests for the early detection and surveillance of CRC.

Mobilizing forces - CD4^＋ helper T cells script adaptive immunity

Traditionally, CD4^＋ T cells have been understood to play a key role in‘helping＇ CD8^＋ T cells undergo efficient activation and proliferation in response to foreign pathogens. This has been thought to be directed primarily by CD4^＋ T cell interactions with dendritic cells （DCs） [1, 2] that convert ‘unlicenced＇ DCs into DCs capable of imple-menting a full blown immune response （‘licenced＇ DCs）.

乳糜泻患者麸质激发试验后外周血T细胞的改变

Background: Current understanding of T cell epitopes in coeliac disease (CD) largely derives from intestinal T cell clones in vitro. T cell clones allow identification of gluten peptides that stimulate T cells but do not quantify their contribution to the overall gluten specific T cell response in individuals with CD when exposed to gluten in vivo. Aims: To determine the contribution of a putative dominant T cell epitope to the overall gliadin T cell response in HLA- DQ2 CD in vivo. Patients: HLA- DQ2+ individuals with CD and healthy controls. Methods: Subjects consumed 20 g of gluten daily for three days. Interferon γ (IFN- γ ) ELISPOT was performed using peripheral blood mononuclear cells (PBMC) to enumerate and characterise peptide and gliadin specific T cells before and after gluten challenge. Results: In 50/59 CD subjects, irrespective of homo- or heterozygosity for HLA- DQ2, IFN- γ ELISPOT responses for an optimal concentration of A- gliadin 57- 73 Q- E65 were between 10 and 1500 per million PBMC, equivalent to a median 51% of the response for a “ near optimal" concentration of deamidated gliadin. Whole deamidated gliadin and gliadin epitope specific T cells induced in peripheral blood expressed an intestinal homing integrin (α 4β 7) and were HLA- DQ2 restricted. Peripheral blood T cells specific for A- gliadin 57- 73 Q- E65 are rare in untreated CD but can be predictably induced two weeks after gluten exclusion. Conclusion: In vivo gluten challenge is a simple safe method that allows relevant T cells to be analysed and quantified in peripheral blood by ELISPOT, and should permit comprehensive high throughput mapping of gluten T cell epitopes in large numbers of individuals with CD.

乳糜泻小麦麸质蛋白的显性T细胞表位的拮抗剂及其无毒性变异

Background:Coeliac disease(CD)is due to an inappropriate T cell mediated response to specific gluten peptides.Measured by interferon γ(IFN-γ)ELISPOT,about half of the gliadin specific T cells induced with in vivo wheat gluten exposure in HLA-DQ2+CD are specific for an α/β-gliadin peptide(p57-73 QE65;QLQPFPQPELPYPQ-PQS)that includes two overlapping T cell epitopes(PFPQPELPY and PQPELPYPQ).Aim:To define minimally substituted variants of p57-73 QE65 universally devoid of IFN-γstimulatory capacity but capable of antagonising IFN-γsecretion from polyclonal T cells specific for p57-73 QE65.Methods:Peripheral blood mononuclear cells collected from 75 HLA-DQ2+CD patients after in vivo gluten challenge were used in overnight ELISPOT assays to screen 218 single or double substituted variants of p57-73 QE65 for cytokine stimulatory and antagonist activity.Results:The region p60-71(PFPQPELPYPQP)and especially p64-67(PELP)was sensitive to substitution.Twelve substitutions in p64-67 stimulated no IFN-γELISPOT response.Among 131 partial agonists identified,45 produced statistically significant inhibition of IFN-γELISPOT responses when cocultured in fivefold excess with p57-73 QE65(n = 10).Four substituted variants of p57-73 QE65 were inactive by IFN-γELISPOT but consistently antagonised IFN-γELISPOT responses to p57-73 QE65,and also retained interleukin 10 stimulatory capacity similar to p57-73 QE65.Conclusions:Altered peptide ligands of p57-73 QE65,identified using polyclonal T cells from multiple HLA-DQ2+CDdonors,have properties in vitro that suggest that a single substitution to certain α/β-gliadins could abolish their capacity to stimulate IFN-γfrom CD4 T cells and also have anti-inflammatory or protective effects in HLA-DQ2+CD.

From signalling pathways to targeted therapies: unravelling glioblastoma’s secrets and harnessing two decades of progress

Glioblastoma,a rare,and highly lethal form of brain cancer,poses significant challenges in terms of therapeutic resistance,and poor survival rates for both adult and paediatric patients alike.Despite advancements in brain cancer research driven by a technological revolution,translating our understanding of glioblastoma pathogenesis into improved clinical outcomes remains a critical unmet need.This review emphasises the intricate role of receptor tyrosine kinase signalling pathways,epigenetic mechanisms,and metabolic functions in glioblastoma tumourigenesis and therapeutic resistance.We also discuss the extensive efforts over the past two decades that have explored targeted therapies against these pathways.Emerging therapeutic approaches,such as antibody-toxin conjugates or CAR T cell therapies,offer potential by specifically targeting proteins on the glioblastoma cell surface.Combination strategies incorporating protein-targeted therapy and immune-based therapies demonstrate great promise for future clinical research.Moreover,gaining insights into the role of cell-of-origin in glioblastoma treatment response holds the potential to advance precision medicine approaches.Addressing these challenges is crucial to improving outcomes for glioblastoma patients and moving towards more effective precision therapies.

Ndfip1 represses cell proliferation by controlling Pten localization and signaling specificity

Pten controls a signaling axis that is implicated to regulate cell proliferation,growth,survival,migration,and metabolism.The molecular mechanisms underlying the specificity of Pten responses to such diverse cellular functions are currently poorly understood.Herewe report the control of Pten activity and signaling specificity during the cell cycle by Ndfip1 regulation of Pten spatial distribution.Genetic deletion of Ndfip1 resulted in a loss of Pten nuclear compartmentalization and increased cell proliferation,despite cytoplasmic Pten remaining active in regulating PI3K/Akt signaling.Cells lacking nuclear Pten were found to have dysregulated levels of Plk1 and cyclin D1 that could drive cell proliferation.In vivo,transgene expression of Ndfip1 in the developing brain increased nuclear Pten and lengthened the cell cycle of neuronal progenitors,resulting in microencephaly.Our results show that local partitioning of Pten from the cytoplasm to the nucleus represents a key mechanism contributing to the specificity of Pten signaling during cell proliferation.

Transcription tipping points for T follicular helper cell and T-helper 1 cell fate commitment

During viral infection,immune cells coordinate the induction of inflammatory responses that clear infection and humoral responses that promote protection.CD4^(+)T-cell differentiation sits at the center of this axis.Differentiation toward T-helper 1(Th1)cells mediates inflammation and pathogen clearance,while T follicular helper(Tfh)cells facilitate germinal center(GC)reactions for the generation of high-affinity antibodies and immune memory.While Th1 and Tfh differentiation occurs in parallel,these CD4^(+)T-cell identities are mutually exclusive,and progression toward these ends is determined via the upregulation of T-bet and Bcl6,respectively.These lineage-defining transcription factors act in concert with multiple networks of transcriptional regulators that tip the T-bet and Bd6 axis in CD4^(+)T-cell progenitors to either a Th1 or Tfh fate.It is now clear that these transcriptional networks are guided by cytokine cues that are not only varied between distinct viral infections but also dynamically altered throughout the duration of infection.Thus,multiple intrinsic and extrinsic factors combine to specify the fate,plasticity,and function of Th1 and Tfh cells during infection.Here,we review the current information on the mode of action of the lineage-defining transcription factors Bcl6 and T-bet and how they act individually and in complex to govern CD4^(+)T-cell ontogeny.Furthermore,we outline the multifaceted transcriptional regulatory networks that act upstream and downstream of Bcl6 and T-bet to tip the differentiation equilibrium toward either a Tfh or Th1 fate and how these are impacted by dynamic inflammatory cues.

The network of cytokines,receptors and transcription factors governing the development of dendritic cell subsets

The pathways leading to the development of different dendritic cell(DC)subsets have long been unclear.In recent years,a number of precursors on the route to DC development,both under steady state and inflammatory conditions,have been described,and the nature of these pathways is becoming clearer.In addition,the development of various knockout mouse models and an in vitro system modelling DC development have revealed the role of numerous cytokines and transcription factors that influence DC development.Here,we review recent findings on the factors important in DC development in the context of the developmental pathways that have been described.

Discussion of some‘knowns’and some‘unknowns’about the tumour suppressor p53

Activation of the tumour suppressor p53 upon cellular stress can induce a number of different cellular processes?The diverse actions of these processes are critical for the protective function of p53 in preventing the development of cancer.However,it is still not fully understood which process(es)activated by p53 is/are critical for tumour suppression and how this might differ depending on the type of cells undergoing neoplastic transformstion and the nature of the drivers of oncogenesis.Moreover,it is not clear why upon activation of p53 some cells undergo cell cycle arrest and senescence whereas others die by apoptosis.Here we discuss some of the cellular processes that are crucial for p53-mediated tumour suppression and the factors that could impact cell fate upon p53 activation.Finally,we describe therapies aimed either at activating wild-type p53 or at changing the behaviour of mutant p53 to unleash tumour growth suppressive processes for therapeutic benefit in malignant disease.

Csk-homologous kinase （Chk/Matk）： a molecular policeman suppressing cancer formation and progression

Aberrant activation of Src-family tyrosine kinases （SFKs） directs initiation of metastasis and development of drug resistance in multiple solid tumors and hematological cancers. Since oncogenic mutations of SFKs are rare events, aberrant activation of SFKs in cancer is likely due to dysregulation of the two major upstream inhibitors： C-terminal Src kinase （Csk） and its homolog Csk-homologous kinase （Chk/Matk）. Csk and Chk/Matk inhibit SFKs by selectively phosphorylating the inhibitory tyrosine residue at their C-terminal tail. Additionally, Chk/Matk can also employ a non- catalytic inhibitory mechanism to inhibit multiple active forms of SFKs, suggesting that Chk/Matk is a versatile inhibitor capable of constraining the activity of multiple active forms of SFKs. Mounting evidence suggests that Chk/ Mark is a potential tumor suppressor downregulated by epigenetic silencing and/or missense mutations in several cancers such as colorectal and lung carcinoma. In spite of the potential significance of Chk/Matk in cancer, little is known about its structure and regulation. This review focuses on the mechanisms by which Chk/Matk expression and activity is downregulated in cancers. Specifically, we assessed the evidence demonstrating downregulation of Chk/Matk by epigenetic silencing and missense mutations in cancers. The other focus is the tumor suppressive mechanism of Chk/ Matk. The final focus of the review is on the clinical applications of the investigations into the mechanism of epigenetic silencing of Chk/Matk expression and the tumor suppressive mechanism of Chk/Matk; specifically we discussed how they can benefit the development of biomarkers for early diagnosis of cancers and specific SFK inhibitors for use as cancer therapeutics.

The equivalents of human blood and spleen dendritic cell subtypes can be generated in vitro from human CD34 ＋ stem cells in the presence of fins-like tyrosine kinase 3 ligand and thrombopoietin

Dendritic cells （DCs） are immune cells specialized to capture, process and present antigen to T cells in order to initiate an appropriate adaptive immune response. The study of mouse DC has revealed a heterogeneous population of cells that differ in their development, surface phenotype and function. The study of human blood and spleen has shown the presence of two subsets of conventional DC including the CDlb/c＋ and CD141＋CLEC9A＋ conventional DC （cDC） and a plasmacytoid DC （pDC） that is CD304＋CD123＋. Studies on these subpopulations have revealed phenotypic and functional differences that are similar to those described in the mouse. In this study, the three DC subsets have been generated in vitrofrom human CD34＋ precursors in the presence of fins-like tyrosine kinase 3 ligand （FIt3L） and thrombopoietin （TPO）. The DC subsets so generated, including the CDlb/c＋ and CLEC9A＋ cDCs and CD123 ＋ pDCs, were largely similar to their blood and spleen counterparts with respect to surface phenotype, toll-like receptor and transcription factor expression, capacity to stimulate T cells, cytokine secretion and cross-presentation of antigens. This system may be utilized to study aspects of DC development and function not possible in vivo.

Tranexamic acid for intracerebral haemorrhage within 2 hours of onset: protocol of a phase Ⅱ randomised placebo-controlled double-blind multicentre trial

Rationale Haematoma growth is common early after intracerebral haemorrhage(ICH),and is a key determinant of outcome.Tranexamic acid,a widely available antifibrinolytic agent with an excellent safety profile,may reduce haematoma growth.Methods and design Stopping intracerebral haemorrhage with tranexamic acid for hyperacute onset presentation including mobile stroke units(STOP-MSU)is a phase Ⅱ double-blind,randomised,placebo-controlled,multicentre,international investigator-led clinical trial,conducted within the estimand statistical framework.Hypothesis In patients with spontaneous ICH,treatment with tranexamic acid within 2 hours of onset will reduce haematoma expansion compared with placebo.Sample size estimates A sample size of 180 patients(90 in each arm)would be required to detect an absolute difference in the primary outcome of 20%(placebo 39%vs treatment 19%)under a two-tailed significance level of 0.05.An adaptive sample size re-estimation based on the outcomes of 144 patients will allow a possible increase to a prespecified maximum of 326 patients.Intervention Participants will receive 1 g intravenous tranexamic acid over 10 min,followed by 1 g intravenous tranexamic acid over 8 hours;or matching placebo.Primary efficacy measure The primary efficacy measure is the proportion of patients with haematoma growth by 24±6 hours,defined as either≥33%relative increase or≥6 mL absolute increase in haematoma volume between baseline and follow-up CT scan.Discussion We describe the rationale and protocol of STOP-MSU,a phase Ⅱ trial of tranexamic acid in patients with ICH within 2 hours from onset,based in participating mobile stroke units and emergency departments.

Virology at the Lorne Infection and Immunity Conference 2019

The Lome Infection and Immunity Conference is one of five scientific meetings held during each month of February at the Cumberland resort in the picturesque seaside town of Lome,on the Great Ocean Road in Victoria(Australia).The specific aim of the meeting is to bring together basic,clinical and translational researchers-those who examine microbes and their impact on the innate or adaptive immune response,researchers who study the mechanisms that regulate immune responses,and those who apply this knowledge to preventing and treating infectious and in”ammatory diseases.The average number of attendees is 220,with registrants appreciative of the welcoming and relaxed atmosphere(Fig.1).

Altered energy metabolism and metabolic gene expression associated with increased metastatic capacity identified in MDA -MB-231 cell line variants

Aim:Despite current advances in therapies and the gradual decline in breast cancer-related mortality,metastasis remains a major therapeutic challenge for treatment.Energy reprogramming is now recognized to be an important part of tumorigenic processes,but its relevance in metastatic dissemination has yet to be elucidated.Methods:Using the MDA-MB-231HM.LNm5 cell line,a novel,highly metastatic variant line derived from TN hu-man breast adenocarcinoma MDA-MB-231 line,alteration in growth and energy metabolisms associated with en-hanced metastatic potential were described.Glycolysis and oxidative phosphorylation(OXPHOS)was character-ized using the seahorse XF analyzer.Whole transcriptome sequencing(RNA-seq)and quantitative real-time PCR was used to ascertain expression differences in metabolic genes.Results:We observed reduced proliferation,and an elevation of both glycolytic and OXPHOS metabolism in the highly metastatic daughter line.The elevated metabolic rate is only partially reflected by transcript levels of rel-evant metabolic regulators.Heightened mitochondrial respiration is potentially underpinned by increased expres-sion mitochondrial electron transport chain components.However,increased glycolysis was not underpinned by up-regulation of metabolic genes encoding enzymes participating in glycolysis.Conclusion:Our results indicate breast tumour cells with elevated metastatic propensity are more metabolic ac-tive.We also identified differentially expressed metabolic genes,such as IDH2,that may play a part in the meta-static process beyond energy reprogramming.

Promises and pitfalls of targeted agents in chronic lymphocytic leukemia

Targeted agents have significantly improved outcomes for patients with chronic lymphocytic leukemia,particularly high-risk subgroups for whom chemoimmunotherapy previously offered limited efficacy.Two classes of agent in particular,the Bruton tyrosine kinase inhibitors(e.g.,ibrutinib)and the B-cell lymphoma 2 inhibitor,venetoclax,induce high response rates and durable remissions in the relapsed/refractory and frontline settings.However,maturing clinical data have revealed promises and pitfalls for both agents.These drugs induce remissions and disease control in the majority of patients,often in situations where modest efficacy would be expected with traditional chemoimmunotherapy approaches.Unfortunately,in the relapsed and refractory setting,both agents appear to be associated with an inevitable risk of disease relapse and progression.Emerging patterns of resistance are being described for both agents but a common theme appears to be multiple sub-clonal drivers of disease progression.Understanding these mechanisms and developing effective and safe methods to circumvent the emergence of resistance will determine the longer-term utility of these agents to improve patients’quality and length of life.Rational drug combinations,optimised scheduling and sequencing of therapy will likely hold the key to achieving these important goals.

CIS controls the functional polarization of GM-CSF-derived macrophages

The cytokine granulocyte-macrophage-colony stimulating factor (GM-CSF) possesses the capacity to differentiate monocytes into macrophages (MØs) with opposing functions, namely, proinflammatory M1-like MØs and immunosuppressive M2-like MØs. Despite the importance of these opposing biological outcomes, the intrinsic mechanism that regulates the functional polarization of MØs under GM-CSF signaling remains elusive. Here, we showed that GM-CSF-induced MØ polarization resulted in the expression of cytokine-inducible SH2-containing protein (CIS) and that CIS deficiency skewed the differentiation of monocytes toward immunosuppressive M2-like MØs. CIS deficiency resulted in hyperactivation of the JAK-STAT5 signaling pathway, consequently promoting downregulation of the transcription factor Interferon Regulatory Factor 8 (IRF8). Loss- and gain-of-function approaches highlighted IRF8 as a critical regulator of the M1-like polarization program. In vivo, CIS deficiency induced the differentiation of M2-like macrophages, which promoted strong Th2 immune responses characterized by the development of severe experimental asthma. Collectively, our results reveal a CIS-modulated mechanism that clarifies the opposing actions of GM-CSF in MØ differentiation and uncovers the role of GM-CSF in controlling allergic inflammation.

Tackling global health security by building an academic community for One Health action

Background One Health approach is crucial to tackling complex global public health threats at the interface of humans, animals, and the environment. As outlined in the One Health Joint Plan of Action, the international One Health community includes stakeholders from different sectors. Supported by the Bill & Melinda Gates Foundation, an academic community for One Health action has been proposed with the aim of promoting the understanding and real-world implementation of One Health approach and contribution towards the Sustainable Development Goals for a healthy planet.Main text The proposed academic community would contribute to generating high-quality scientific evidence, distilling local experiences as well as fostering an interconnected One Health culture and mindset, among various stakeholders on different levels and in all sectors. The major scope of the community covers One Health governance, zoonotic diseases, food security, antimicrobial resistance, and climate change along with the research agenda to be developed. The academic community will be supported by two committees, including a strategic consultancy committee and a scientific steering committee, composed of influential scientists selected from the One Health information database. A workplan containing activities under six objectives is proposed to provide research support, strengthen local capacity, and enhance global participation.Conclusions The proposed academic community for One Health action is a crucial step towards enhancing communication, coordination, collaboration, and capacity building for the implementation of One Health. By bringing eminent global experts together, the academic community possesses the potential to generate scientific evidence and provide advice to local governments and international organizations, enabling the pursuit of common goals, collaborative policies, and solutions to misaligned interests.

Development of Dendritic Cell System

The dendritic cell system contains conventional dendritic cells(DCs)and plasmacytoid pre-dendritic cells (pDCs).Both DCs and pDCs are bone marrow derived cells.Although the common functions of DCs are antigen-processing and T-lymphocyte activation,they differ in surface markers,migratory patterns,and cytokine output.These differences can determine the fate of the T cells they activate.Several subsets of mature DCs have been described in both mouse and human and the developmental processes of these specialized DC subsets have been studied extensively.The original concept that all DCs were of myeloid origin was questioned by several recent studies,which demonstrated that in addition to the DCs derived from myeloid precursors, some DCs could also be efficiently generated from lymphoid-restricted precursors.Moreover,it has been shown recently that both conventional DCs and pDCs can be generated by the Flt3 expressing hemopoietic pro- genitors regardless of their myeloid-or lymphoid-origin.These findings suggest an early developmental flexibility of precursors for DCs and pDCs.This review summarizes some recent observations on the deve- lopment of DC system in both human and mouse.Cellular & Molecular Immunology.2004;1(2):112-118.