In addition, 18F-FDG PET can be used to investigate functional connectivity based on changes in glucose metabolism, which reflects local brain activity. Previous studies have highlighted the feasibility and clinical usefulness of 18F-FDG-PET for the analysis of RSNs, and recent studies have utilized simultaneous PET/fMRI for such analyses. While PET provides seed information regarding the focus of the abnormalities , fMRI is used for the analysis of functional connectivity. Thus, as PET and fMRI provide different types of information, integrating these modalities may aid in elucidating the pathological mechanisms underlying certain diseases, and in characterizing individual patients.Resting-state networks (RSNs) exhibit spontaneous functional connectivity in the resting state. Previous studies have evaluated RSNs mainly based on spontaneous fluctuations in blood oxygenation level-dependent (BOLD) signals during functional magnetic resonance imaging (fMRI). However, separation between regional increases in cerebral blood flow (CBF) and oxygen consumption is theoretically difficult using BOLD-fMRI. Such separation can be achieved using quantitative  Resting-state networks (RSNs) exhibit spontaneous functional connectivity in the resting state. Among these is the default mode network (DMN), in which the posterior cingulate cortex (PCC) plays a central role, connecting with the medial-prefrontal, hippocampal, and lateral temporal areas . While t2) to cerebral blood flow (CBF). Increases in CBF during task-related brain activation occur in conjunction with smaller changes in CMRO2, resulting in a relative decrease in the OEF. In contrast, the OEF increases during the deactivated state. The default mode network was first discovered by Raichle et al. using positron emission tomography (PET) (2001). The authors focused on the regional oxygen extraction fraction (OEF), and which refers to the ratio of the cerebral metabolic rate of oxygen  to analyze functional connectivity based on spontaneous fluctuations in blood oxygenation-level dependent (BOLD) signals . When an2 . TherefoIn the present study, we summarized recent advances in the evaluation of functional connectivity using PET. Since the methodological aspects of connectivity analysis have been summarized by 2 can be quantitatively evaluated via 15O-labeled gas and water PET  following intravenous injection of FDG, and fluctuations among these frames were evaluated based on global normalization. n = 30), left-sided TLE , and healthy controls (n = 31). FDG-PET has also been used in clinical practice to detect seizure lateralization in patients with TLE. Indeed, patients with RTLE exhibit higher lobar connectivity weights than those with LTLE for connections involving the temporal and parietal lobes of the contralateral hemisphere. Although the mechanisms underlying such differences remain to be fully elucidated, the authors suspected that compensatory mechanisms were more prominent in patients with RTLE than in those with LTLE.Glucose is the fundamental metabolic substrate in the brain. Given that glucose metabolism reflects local brain activity, this measure can be used to analyze functional connectivity. 18F-FDG PET data, revealing that patients with AD exhibit gradual disruptions in functional brain connectivity during the progression of cognitive decline. 18F-FDG PET.18F-FDG-PET analyses of functional connectivity are clinically useful for identifying potential responders to therapy.Another study highlighted the usefulness of functional connectivity analysis for predicting responses to vagus nerve stimulation (VNS) in patients with epileptic seizures . Yu et a18F-FDG-PET, observing that temporal changes in glucose metabolism can be evaluated during continuous injection of FDG. Since FDG is metabolically trapped in neurons or glial cells in the brain after being metabolized into glucose-6-phosphate by hexokinase, the upslope of the time-activity curves reflects real-time glucose metabolism. This technique can used to detect focal brain regions exhibiting task- or stimulus-related activation, similar to BOLD-fMRI . Similar networks were detected using the two modalities, suggesting that coupling between glucose metabolism and blood oxygenation responses is preserved in healthy individuals. 18F-FDG-PET was used to detect regions exhibiting hypometabolism, rather than for connectivity analysis. Within-subject analysis, in which correlations between glucose utilization (PET) and connectivity (fMRI) were evaluated for each participant, revealed that patients with MCI/AD exhibited significant decreases in such correlations. For between-subjects comparisons of PET/fMRI data, in which each imaging modality and metric (volumes) were concatenated across all participants, overlap between the two modalities varied depending on fMRI connectivity parameters.Recent studies have utilized simultaneous or combined PET/MRI for functional connectivity analysis, allowing for perfect co-registration in terms of spatial, and temporal matching . Using aPrevious studies have compared the correlations between PET and fMRI data using simultaneous PET/MRI. 18F-THK5351 PET in amyloid-positive patients with early AD (n = 23) and healthy, amyloid-negative controls (n = 24). To perform seed-based connectivity analysis, they generated several seed ROIs based on significant differences in 18F-THK5351 retention between the AD and control groups. Histological evaluations at autopsy indicated that 18F-THK5351 retention corresponded to tau deposition, monoamine oxidase-B (MAO-B) levels, and astrogliosis in the brains of patients with AD. Furthermore, seed-based connectivity analysis revealed significant decreases in connectivity between the posterior cingulate cortex (PCC) and widespread brain regions in patients with AD.11C]ABP688 PET analysis in patients with major depression. To determine metabotropic glutamate receptor-5 (mGluR5) availability, the binding potential (BPND) of [11C]ABP688 was quantified using the simplified reference tissue model. ROI-based analysis revealed that the [11C]ABP688 BPND in the prefrontal cortex was significantly lower in patients than in controls. BPND seed-based functional connectivity analysis revealed significantly reduced connectivity between the inferior parietal cortex and fusiform gyrus/inferior occipital cortex in patients, relative to that observed in controls. Patterns of correlation between [11C]ABP688 BPND and the strength of functional connectivity involving the superior prefrontal cortex seed were opposite in the depression and control groups. Thus, using PET to determine mGluR5 availability revealed related alterations in functional connectivity in the patient group. In addition, ND differences on 11C-raclopride PET as seeds. They examined dopamine D2 receptors using 11C-raclopride PET in patients with major depressive disorder (n = 16) and controls (n = 14). Increased BPND in both the left ventral striatum and right dorsal striatum and decreased functional connectivity between the affected striatal areas and their respective cortical targets were observed in patients with major depressive disorder.Although previous reports on functional connectivity determined using simultaneous PET/MR are still limited , a combi15O-gas and water PET. Glucose metabolism during 18F-FDG PET can also be used for functional connectivity analysis. Moreover, PET can provide information regarding the focus of abnormalities, which can then be used for functional connectivity analyses via fMRI. Comparisons between PET and fMRI data may thus allow for improved understanding of the pathophysiology of several brain disorders. Such comparisons may also aid in characterizing individual patients with these disorders.Functional correlations between blood flow and oxygen consumption can be separately estimated using quantitative Both authors wrote and approved the final version of the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Vascular inflammation evaluated by FDG PET has been positively related to metabolic syndrome components and traditional risk factors of cardiovascular disease, including high-sensitivity C-reactive protein, body mass index, and insulin resistance. A positive association of vascular inflammation with endothelial dysfunction, resistin levels, pericardial adipose tissue, and visceral fat area has also been reported. In contrast, HDL cholesterol and adiponectin have been inversely related to vascular inflammation detected by FDG PET. Because of its reproducibility, serial FDG PET shows potential for tracking the effects of dietary interventions and other systemic and local antiatherosclerotic therapies for plaque inflammation.Vascular inflammation plays a central role in atherosclerosis, from initiation and progression to acute thrombotic complications. Modified low-density lipoproteins (LDLs) and apoB-containing particles stimulate plaque inflammation by interacting with macrophages. Loss of function of high-density lipoprotein (HDL) for preventing LDL particles from oxidative modification in dyslipidemic states may amplify modified LDL actions, accelerating plaque inflammation. Diets are one of the most important factors that can affect these processes of lipoprotein oxidation and vascular inflammation. Recently,  The main aim of this review is to describe the interactions between lipoprotein and macrophages leading to vascular inflammation during atherosclerosis, effects of diets during this process of inflammation, and evaluation of this vascular inflammation by 18F- FDG PET. Atherosclerotic cardiovascular disease (ASCVD), manifested as various forms of fatal diseases including myocardial infarction, ischemic stroke, and peripheral artery occlusive disease, is the leading cause of morbidity and mortality . TherefoInflammation is known to play a key role in the development and progression of atherosclerosis , and athActivated macrophages are among the major contributors to increased susceptibility of plaque rupture and promoting thrombus formation . MacrophElevated circulating inflammatory markers, particularly C-reactive protein (CRP), can predict the risk of atherosclerotic cardiovascular events ,8 and poData from experimental and clinical studies indicate that statins not only lower low-density lipoproteins (LDLs) but also reduce plaque inflammation and affect plaque stability ,9. MacroCRP testing may be useful for targeted statin therapy, particularly for the primary prevention of cardiovascular events . In the Modified LDL and other apoB-containing particles represent key stimulators of plaque inflammation through interplay with macrophages  Figure . The endLoss of the function of HDL to protect LDL particles from oxidative modification in dyslipidemic states, including hypertriglyceridemia, may amplify the role of modified LDL and accelerate intraplaque inflammation  Figure . HDL parEffect of diets on LDL oxidation and inflammation has been consistently reported ,5 and suIncreasing evidences indicate that induction or alleviation of a proinflammatory condition and subsequent alteration in endothelial function by dietary patterns may be one mechanism that CVD risks are affected by diets . IngestiFurthermore, some animal studies have tried to evaluate the effect of dietary modification on vascular inflammation and plaque contents of atherosclerotic lesions more directly. Verhamme et al.  reportedIn clinical testing, there have been trials determining the long-term effect of dietary intervention on markers of vascular inflammation and plaque stability. A 12-month randomized trial with 164 individuals at high risk for CVD showed that enhanced Mediterranean diets attenuated the biomarkers related to atherosclerotic vascular inflammation and plaque vulnerability, including adhesion molecule expression on monocyte surface, CRP, interleukin-6, and endothelial adhesion molecules, compared to a low-fat diet . Accordi18F-FDG PET has been established as one of the most useful imaging modalities for identifying and measuring inflamed vulnerable atherosclerotic plaques [18F-FDG PET is generally used in clinical fields to detect 18F-FDG uptake in cells with increased glucose metabolism [18F-FDG accumulation is also increased by active atherosclerosis [18F-FDG uptake on PET images [18F-FDG PET is attributed to the high glycolytic activity of macrophages within active atherosclerotic plaques [18F-FDG PET in eight patients followed by carotid endarterectomy. Rogers et al. [18F-FDG PET showed a higher uptake within the ascending aorta, left main coronary artery, and the culprit lesion in 10 patients with recent acute coronary syndrome compared to 15 individuals with stable angina. In the preclinical testing, Ogawa et al. [18F-FDG accumulation in atherosclerotic lesions was ascribable to macrophages in a rabbit model of atherosclerosis. According to Tawakol et al. [18F-FDG PET signals from carotid plaques were significantly correlated with macrophage staining of corresponding pathologic specimens isolated after endarterectomy. These studies indicate that 18F-FDG PET is a useful noninvasive tool for detecting and quantifying inflammation within atherosclerotic lesions, particularly macrophage-associated vascular inflammation [Recently,  plaques . 18F-FDGtabolism , includitabolism . It has clerosis ,58,59. IT images ,61. Visu plaques ,62  flow-mediated dilation of the brachial arteries in 145 adults [Honda et al. confirmed that vascular inflammation in carotid arteries assessed by 5 adults . Endothe5 adults . The association between vascular inflammation and serum adiponectin or resistin levels has also been investigated. Adiponectin is a metabolically active adipokine  that exe18F-FDG PET in 93 men and women without diabetes or CVD [Additionally, pericardial adipose tissue and visceral fat area were shown to be associated with vascular inflammation measured by s or CVD . Pericars or CVD .18F-FDG uptake by atherosclerotic plaques is considered a transient phenomenon that is attenuated according to decreases in active inflammatory components [18F-FDG uptake by aortas, whereas intimal thickening was not altered in Watanabe heritable hyperlipidemic rabbits.Vascular inflammation may denote a dynamic aspect of atherosclerotic lesions, which varies during the course of atherosclerosis . 18F-FDGmponents . Clinicamponents ,60,83,84mponents . Ogawa e18F-FDG PET/CT in atherosclerotic rabbits. In the same study, immunoreactivity for macrophages and oxidized phospholipids from the aortas showed a significantly decreased value in the pioglitazone group compared to that in control rabbits [PPAR-gamma receptors are expressed by cells involved in the atherosclerotic process, including endothelial cells, smooth muscle cells, T-lymphocytes, and primarily macrophages ,87. In a rabbits .18F-FDG uptakes in FDG-PET in humans. In the simvastatin group, the reduction in 18F-FDG accumulation was well correlated with the increase in HDL cholesterol but not with the decrease in LDL cholesterol [Tahara et al.  reportedlesterol , suggestlesterol . 18F-FDG uptake in the aorta was significantly lowered by a cholesterol-lowering diet [18F-FDG uptake in PET/CT in 60 healthy adults, and the extent of reduction in the 18F-FDG index correlated well with elevations in plasma HDL cholesterol levels [When analyzed with in vivo PET/CT scan and ex vivo gamma counting of excised aorta in hypercholesterolemic mice deficient in LDL receptor and expressing only apolipoprotein B-100, ing diet . At the ing diet . Notablying diet  reportedl levels . Lifestyl levels  included18F-FDG PET for monitoring the effects of antiatherosclerotic therapies, including dietary intervention on vascular inflammation. Serial 18F-FDG PET may be a noninvasive tool for tracking the effects of therapeutic interventions on plaque inflammation and developing novel drugs that can recede inflammation on vulnerable plaques [The results of previous studies suggest the utility of  plaques ,60,83,8418F-FDG PET has emerged as a reliable imaging technique capable of identifying vascular inflammation associated with macrophage infiltration. Clinical studies suggest the utility of 18F-FDG PET for monitoring effects of dietary interventions and other antiatherosclerotic therapies on plaque inflammation. Although experimental and clinical studies provide indirect evidences that vascular inflammation may be affected by diets, further clinical studies addressing the effect of diet directly on atherosclerotic vascular inflammation and plaque vulnerability are needed. 18F-FDG PET can be a useful tool in identifying this issue.An increasing body of evidence supports that vascular inflammation, which is stimulated by modified LDLs and apoB-containing particles, and their interplay with macrophages plays a key action in atherosclerosis.


Our ultimate aim is to provide a comprehensive summary of the currently available information that will encourage and facilitate further mechanistic elucidation of iron-induced pathoelectrophysiological changes in the heart.Iron, the most abundant transition metal element in the human body, plays an essential role in many physiological processes. However, without a physiologically active excretory pathway, iron is subject to strict homeostatic processes acting upon its absorption, storage, mobilization, and utilization. These intricate controls are perturbed in primary and secondary hemochromatoses, leading to a deposition of excess iron in multiple vital organs including the heart. Iron overload cardiomyopathy is the leading cause of mortality in patients with iron overload conditions. Apart from mechanical deterioration of the siderotic myocardium, arrhythmias reportedly contribute to a substantial portion of cardiac death associated with iron overload. Despite this significant impact, the cellular mechanisms of electrical disturbances in an iron-overloaded heart are still incompletely characterized. This review article focuses on cellular electrophysiological studies that directly investigate the effects of iron overload on the function of cardiac ion channels, including trans-sarcolemmal and sarcoplasmic reticulum Ca Iron is an essential metal element that serves many physiological functions. These range from oxygen transport by hemoglobin to catalytic activities by various iron-containing enzymes. Requirement of iron for life can also be illustrated by the ability of the immune system to restrict microbial invasion by iron-sequestrating strategies . Since tCardiac iron deposition is markedly heterogenous and preferentially occurs in the basal segments of the anterior and inferior ventricular walls . Patchy Iron-overloaded hearts exhibits gradual deterioration in both mechanical function and electrical activity. Early detectable arrhythmias include occasional premature atrial/ventricular contractions and first-degree atrioventricular (AV) block, whereas sinoatrial node dysfunction, atrial flutter/fibrillation, second- and third-degree AV block, and ventricular tachycardia/fibrillation tend to occur in advanced stages of the disease . Repolar40 and APD80) remained unchanged ; however, the maximum diastolic potential and the slope of rapid depolarization (phase 0) were unaffected  in iron-overloaded rat LV cardiomyocytes (Supplementary Table nchanged . Howeveruced APA . Consist APA (50 . In mousaffected . Investiaffected  and fromaffected . Howeveraffected . Intersp2O2), the results may be considered to represent the downstream effects of iron-induced ROS-mediated cellular damage on the electrical properties of the cardiomyocyte. Interestingly, ROS-induced afterdepolarizations have been mechanistically linked to overactivation of Ca2+/calmodulin kinase (CaMK) II in rabbit ventricular cardiomyocytes (2O2-inducedAP changes (2O2 and superoxide anion (•O2-) to an even more potent ROS, hydroxyl radical (•OH) (Since the generation of reactive oxygen species (ROS) is likely to be a core pathophysiological process of iron-related cellular/organellar injury , researcmyocytes . As expe changes , most lial (•OH) .The precise mechanism for QTc prolongation/dispersion in patients with iron overload remains obscure. Since iron loading was shown to induce APD shortening in various models  have been proposed as candidate molecular entrances into the cardiomyocyte for the ferrous form (Fe2+) of non-transferrin bound iron (NTBI)  . Although the proof-of-concept experiment showing permeation and blockade of the TTCC by Fe2+in CaV3.1-transfected HEK293 cells has been reported  . This poand TTCC . However2+ plus various concentrations of iron, Fe2+, but not Fe3+, reversibly altered kinetic properties of the LTCC. Fe2+ (0.5–4 mM) delayed L-type Ca2+ current  inactivation in a dose-dependent manner  accentuated peak ICa,L and increased net Ca2+ influx via the LTCC, but these parameters were suppressed at higher Fe2+ concentrations (1–4 mM) . Hypothetically, in the face of impaired ICa,L, cellular electrophysiological remodeling that leads to the compensatory upregulation of ICa,T channel may occur. Proving this hypothesis will clarify the relative roles of LTCC and TTCC in cardiac iron-overload conditions, as well as the role of pharmacological modulations of TTCC.In isolated rat LV cardiomyocytes superfused with 2 mM Cat manner , possibltivation . Lower c(1–4 mM) . Even thICa,L  have also been evaluated  compared to CaV1.2-mediated ICa,L (≈-30 mV), and the depolarizing shift of ICa,L current-voltage relationship in iron-overloaded SAN and RAA cardiomyocytes resembled the results described in CaV1.3-deficient mice  in RAA cardiomyocytes from chronically iron-overloaded mice. The attempt to measure the protein levels failed due to a technical error  (2+ ATPase (2+ ATPase (SERCA) (2+exerts its ROS-independent inhibitory effect on ryanodine receptors (RyR) via competitive binding, thus compromising Ca2+-induced Ca2+ release from the sarcoplasmic reticulum  is yet to be systematically proven in iron-overloaded cardiomyocytes.Abnormal trans-sarcolemmal and subcellular Caomyocyte . These ihineries . Excess er (NCX) , sarcole+ ATPase ,b, and s (SERCA) . Moreoveeticulum . The cau+ current (INa) in short-term iron-incubated  cultured rat LV cardiomyocytes and also in freshly isolated LV cardiomyocytes from chronically iron-injected gerbils  was increased, and a small depolarizing shift (≈5 mV) of its inactivation curve was observed (+ current (IK1) was unaffected (Multiple Kobserved . Howeveraffected .+(KATP) channel current (IKATP) could be activated by iron-induced oxidative stress  K+ channels, which mediate the rapid delayed rectifier K+ current (IKr) in the heart, has already been studied in various cell types -sensitive Ke stress . Interesocardium . These fll types . NeverthIh or If) is a major contributor of spontaneous depolarization in SAN cells, Rose et al. reported no significant changes in If density and current-voltage relationship in mice with chronic iron overload  is of no less importance. This point can be exemplified by the complexity and multifaceted nature of intracellular CaThe main purpose of this article is to provide the molecular/cellular basis of iron-induced cardiac pathoelectrophysiology. However, it should be noted that additional data from other aspects are also essential for bridging the gap of knowledge between the cellular phenomena and the clinical phenotypes. These include, but not limited to, alterations in intercellular coupling, roles of non-parenchymal cells , structural remodeling, systemic immunoinflammatory responses, and coincident extracardiac pathology  . Due to + current alterations, amiodarone has been used to treat supraventricular arrhythmias commonly found in iron-overload patients , with identification of potential therapeutic targets and development of practical treatment strategies being the extremely worthwhile goals.NS-A, L-HX searched the literature, collected the data, analyzed the data, and wrote the manuscript. SC, NC studied the design, searched the literature, analyzed the data, and wrote the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


MYD88 p.L265P mutation. Ibrutinib therapy was subsequently commenced, resulting in complete resolution of the skin disease. Despite an ongoing skin response, the patient developed progressive nodal disease at two months. Genomic analysis of the cutaneous tumor sample at baseline was compared to that of the inguinal lymph node upon progression, and revealed the acquisition of multiple genomic changes. These included several aberrations expected to bypass BTK inhibition, including two CARD11-activating mutations, and a deleterious mutation in the nuclear factor kappa B (NF-κB) negative regulator, NFKBIE. In addition, an IgH-IRF8 translocation was detected (which brings the IRF8 transcription factor under control of the immunoglobulin heavy chain locus), representing a third plausible mechanism contributing to ibrutinib resistance. Several copy-number changes occurred in both samples, including an amplification of 18q, which encodes the anti-apoptotic protein BCL2. We describe the first case of novel genomic changes of PCDLBCL-LT that occurred while on ibrutinib, providing important mechanistic insights into both pathogenesis and drug resistance.Primary cutaneous diffuse large B-cell lymphoma, leg type (PCDLBCL-LT) is one of the well-recognized extranodal lymphomas commonly addicted to the B-cell receptor-MYD88 superpathway. We aimed to describe the genomic changes in a patient who progressed through treatment with ibrutinib, a Bruton’s tyrosine kinase (BTK) inhibitor. An 80-year-old woman presented with multiply relapsed PCDLBCL-LT after multiple lines of chemoimmunotherapy and radiotherapy. Pre-treatment testing of the localized cutaneous tumor lesion on a lymphoid amplicon panel demonstrated an  Primary cutaneous diffuse large B-cell lymphoma, leg type (PCDLBCL-LT) is recognized in the World Health Organization classification of hematological malignancies . It predMYD88 p.L265P mutations in patients with PCDLBCL-LT of up to 69% [MYD88 mutations promote the survival of ABC DLBCL cells lines through the nuclear factor kappa B (NF-κB) pathway, which in turn can be blocked through inhibition of Bruton’s tyrosine kinase (BTK) (which links B cell receptor activity to NF-κB transcription). [CD79B and MYD88 co-mutations [Recent studies described rates of p to 69% , where ip to 69% . Activat18F-FDG-PET after 3 cycles. She remained in remission until October 2014, at which point she presented with a lesion on her upper left thigh. A biopsy confirmed relapsed PCDLBCL-LT , and she was subsequently treated with radiotherapy only.We report the case of an 80-year-old woman who presented with a localized subcutaneous nodule in her right axillary fold in May 2012. Histopathology following a core biopsy demonstrated sheets of large cells staining for pan-B cell markers CD5, BCL2, and MUM1, but not for CD10, which in the absence of nodal and extracutaneous disease was diagnostic of PCDLBCL-LT. She received six cycles of R-CHOP chemoimmunotherapy , and attained a complete metabolic response (CMR) on restaging 18F-FDG-PET scan demonstrated the presence of three soft-tissue nodules in her left lower limb. Systemic therapy with rituximab, and reduced-dose gemcitabine and vinorelbine was administered, with subsequent radiotherapy resulting in a partial response. In July 2016, she developed a papule on her left thigh at the margin of the previous radiotherapy field. A third course of radiotherapy was administered, and the patient was commenced on lenalidomide. A further short remission ensued, and in both September and October 2016, new lesions developed outside the previous radiotherapy fields, for which she underwent a fourth course of radiotherapy. An 18F-FDG-PET scan in December 2016 demonstrated complete response in irradiated areas, but also a progression of the disease in her left thigh outside the radiation fields, and nodal progression within the pelvis. She remained in second remission until January 2016, at which point she developed further lesions on her left thigh. Biopsy confirmed relapsed PCDLBCL-LT, and an MYD88 p.L265P missense mutation. More extensive analysis was undertaken using a custom hybridization capture panel developed in-house, comprising 313 genes of interest in hematological malignancies, demonstrating a CD79B p.Y196H mutation.A biopsy of the new cutaneous nodule in her left thigh confirmed PCDLBCL-LT, and for the first time, DNA analysis was performed. Routine testing on a custom-designed lymphoid 26-gene amplicon panel demonstrated an The patient was commenced on ibrutinib (420 mg daily), with subsequent complete resolution of the skin changes. Two months later, she presented with lymphedema in her left leg, and imaging demonstrated a discordant response to ibrutinib, with progressive inguinal, iliac, and paraaortic nodal disease, and an ongoing complete cutaneous response.MYD88 and CD79B mutations. However, additional genetic abnormalities were detected in this post-ibrutinib sample, potentially representing mechanisms of ibrutinib resistance in the progressive nodal disease , a negative regulator of NF-κB, representing a further plausible mechanism of ibrutinib resistance. Normally, NFKBIE levels are upregulated following NF-κB activation, and its usual role is to retain NF-κB in the cytoplasm with subsequent inhibition of NF-κB-directed gene expression [NFKBIE mutation observed in this patient was inactivating, resulting in a lack of cytoplasmic retention of NF-κB, with subsequent increased NF-κB-directed transactivation.A further new abnormality detected in the post-ibrutinib sample was a p.G460A mutation in NFKB inhibitor epsilon  IgH-IRF8), observed in the post-ibrutinib sample only. This translocation brings the IRF8 transcription factor under the control of the immunoglobulin heavy-chain locus, thereby causing its dysregulation. We examined IRF8 expression in both the pre-ibrutinib cutaneous sample and the post-ibrutinib nodal sample . While a, observeTNFRSF11A, which encodes a receptor involved in NF-κB activation via both canonical and non-canonical pathways [CDKN2A, and the other which involved PTRPD, a protein tyrosine phosphatase which acts as a signaling molecule involved in cell-cycle regulation, and oncogenic transformation [Off-target reads from the hybridization capture panel were used to determine low-coverage whole-genome copy-number variation, visualized using a browser developed in-house. This analysis provided insight into disease pathogenesis, with several abnormalities being detected in both the pre- and post-ibrutinib samples . Amplifipathways . Additioormation .The patient underwent further radiotherapy with impressive regression of the left-leg lymphedema. She remained on ibrutinib daily for a further three months with no evidence of recurrence of cutaneous disease; however, she eventually succumbed to progressive disease in the pelvis and abdomen.CD79B, and also an MYD88 p.L265P mutation conferring constitutive activation of NF-κB. These variants were also among a restricted set of recurrent mutations in PCDLBCL-LT, recently identified by Mareschal et al. [CD79B and MYD88 have been reported [MYD88, and wildtype CD79A/B) with ibrutinib and chemotherapy  with remission exceeding 20 months. This patient had neither CARD11 nor NFKBIE mutations detected; however, had mutations in 18 additional genes, several of which  may have affected NF-κB activity [This case has several features consistent with previously understood mechanisms of disease: the pre-ibrutinib sample harbored mutations in both the B-cell receptor (BCR) subunit, Ibrutinib was prescribed until the pre-terminal phase of the illness, and we noted the ongoing cutaneous response to this agent. We believe that this patient exhibited spatial tumoral heterogeneity, with ibrutinib-resistance mutations identified in her nodal disease, but not in her cutaneous disease.We describe the first case of evolving, novel genomic changes in PCDLBCL-LT that occurred while on ibrutinib, providing important mechanistic insights into both pathogenesis, and drug resistance. This highlights the existence of a dynamic mutational landscape, which may limit the efficacy of BTK inhibition in certain patients with this condition.


Recently, pathogenic serum immunoglobulin G (IgG) autoantibodies to myelin oligodendrocyte glycoprotein (MOG) have been detected in a subgroup of patients with central nervous system (CNS) demyelination, including in patients with myelitis. Relatively little is known so far about leptomeningeal involvement in MOG-IgG-positive myelitis. We report the case of a 30-year-old previously healthy woman presenting with longitudinally extensive transverse myelitis and tetraparesis, in whom both the leptomeningeal barrier and the blood-brain barrier (BBB) were altered, as demonstrated by gadolinium-enhanced MRI during relapse. Blood samples taken at onset and four years later were retrospectively found positive for MOG-IgG. Our findings demonstrate that spinal leptomeningeal enhancement (LME) can occur in MOG-IgG-positive encephalomyelitis (EM) and may accompany intraparenchymal BBB breakdown. Pathogenic immunoglobulin G (IgG) autoantibodies directed to myelin oligodendrocyte glycoprotein (MOG), an oligodendrocytic protein localized to the outer surface of the myelin sheaths, have recently been identified in patients with inflammatory CNS demyelination –6. This Myelitis is often associated with leptomeningeal (pia-arachnoid) contrast enhancement (LME). LME is a sign of leptomeningeal barrier impairment and leakage of contrast agent into the subarachnoid space. Understanding LME patterns may help in the differential diagnosis of the various forms of myelitis –10. So fA 30-year-old woman with no previous history of systemic inflammatory disease or neoplastic diseases developed loss of vision in the left eye and two days later in the right eye due to acute ON, followed by tetraparesis two weeks later. Spinal cord MRI obtained prior to treatment revealed LME, intraparenchymal contrast enhancement corresponding to the site of LME, and longitudinal extensive transverse myelitis (LETM) extending from C2 to Th3 .3 with polymorphonuclear predominance; oligoclonal bands were not determined. Aquaporin-4 (AQP4)-IgG was negative. Accordingly, seronegative neuromyelitis optica was suspected by that time. Follow-up MRI demonstrated resolution of LME four months later.Cerebral MRI was normal. Cerebrospinal fluid contained 122 leukocytes/mmRetrospective testing by means of two cell-based assays employing fixed and live HEK293 cells, respectively, transfected with full-length human MOG revealed the presence of MOG-IgG antibodies in a serum sample taken at onset , 2. MOG-Inflammation in demyelinating diseases of the CNS is commonly associated with blood-brain barrier (BBB) disruption. Leptomeningeal involvement has recently been recognized as an important feature in multiple sclerosis pathogenesis . LME has


Recently, we developed a new optogenetic tool, PhotonSABER, which could control AMPA receptor endocytosis in temporal, spatial, and cell-type-specific manners at activated synapses. Using PhotonSABER, we found that AMPA receptor endocytosis and LTD at synapses between parallel fibers and Purkinje cells in the cerebellum mediate oculomotor learning. We also found that PhotonSABER could inhibit endocytosis of epidermal growth factor receptors in HeLa cells upon light stimulation. These results demonstrate that PhotonSABER is a powerful tool for analyzing the physiological functions of endocytosis in non-neuronal cells, as well as the roles of LTD in various brain regions.In the central nervous system, activity-dependent endocytosis of postsynaptic AMPA-type glutamate receptors (AMPA receptors) is thought to mediate long-term depression (LTD), which is a synaptic plasticity model in various neuronal circuits. However, whether and how AMPA receptor endocytosis and LTD at specific synapses are causally linked to learning and memory  In neurons, activity-dependent clathrin-mediated endocytosis of postsynaptic α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)-type glutamate receptors (AMPA receptors) is thought to be the molecular basis of long-term depression (LTD) of synaptic transmission [in vivo remains largely unclear. For example, LTD at synapses between parallel fibers and Purkinje cells is believed to mediate cerebellum-dependent oculomotor learning, such as the adaptation of optokinetic response (OKR) and vestibulo-ocular reflex (VOR) [in vitro slice preparations [in vivo.Clathrin-mediated endocytosis is a cellular process in which cargo-containing clathrin-coated vesicles bud off from the plasma membrane and are transported to early endosomes . This prsmission . Althougex (VOR) . Neverthex (VOR) . Discreparations . Alterna+-adenosine triphosphatase (V-ATPase), chloride channels (ClCs), and Na+/H+ exchangers (NHEs) [+ pumps. Thus, we developed a synthetic H+ pump, termed PhotonSABER, by fusing an engineered Anabaena sensory rhodopsin (ASR) in which Glu has replaced Asp217 (ASRD217E) to direct H+ flow from the extracellular to the cytoplasmic direction, [in vivo.The lumen of endosomes is normally maintained at acidic pH by using the cooperative functions of vacuolar vacuolar-type Hs (NHEs) –11   and endos (NHEs) , we hyporection,  and the rection, , The ligCre driver mice. Light stimulation acutely and reversibly inhibits LTD induction without affecting the normal development of neuronal circuits. For example, LTD at synapses between Schaffer collaterals and CA1 neurons in the hippocampus is proposed to be required for consolidation of context-dependent fear memory, but not for its acquisition [in vivo. The amygdala is believed to be responsible for cue-dependent fear memory. Stimulation of auditory input to the amygdala without foot shock has been shown to induce LTD at these synapses to facilitate the extinction of its associated fear memory [in vivo.PhotonSABER can be used to assess the physiological importance of LTD in various brain regions. PhotonSABER can be expressed in specific cells by mating PhotonSABER knock-in mice, which contain the stop sequence flanked by loxP sites, with brain-region- and cell-type-specific uisition . LTD at uisition . Knock-ir memory . PhotonSNHE6 gene has been identified in Angelman syndrome patients [NHE7 and NHE9 are reported to be linked with Alzheimer’s disease and autism spectrum disorders, respectively [+ to the cytoplasm and pump in K+ into the endosomal lumen. Thus, the endosomal lumen of patients with these diseases may be hyper-acidified . Many growth factors are taken up by cells via endocytosis of growth-factor-bound cell-surface receptors. When HeLa cells were incubated with biotinylated recombinant epidermal growth factor (EGF) for 60 min, EGF signals were detected in Rab7-positive late endosomes. However, when light was applied to HeLa cells expressing PhotonSABER, most EGF signals were not internalized and the internalized signals were not colocalized with Rab7 . AlthougWhy and how does the lumen of early endosomes need to be acidified for their functional maturation? Adenosine diphosphate ribosylation factor nucleotide binding site opener, a guanine exchange factor for the small G proteins of the adenosine diphosphate ribosylation factor family , is repoin vivo.Additional studies are warranted to expand the concept of optogenetic pH regulation of early endosomes to other types of endosomes, such as recycling and late endosomes, to modulate exocytosis and lysosomal trafficking, respectively, and clarify their physiological functions Regarding endocytosis of EGF, HeLa cells expressing hemagglutinin (HA)-tagged PhotonSABER and Rab7-green fluorescent protein were treated with biotinylated EGF  for 60 min in the presence or absence of light stimulation (2000 lx). Cells were fixed with 4% paraformaldehyde in phosphate buffered saline (PBS) for 10 min at room temperature. After being washed with PBS three times, cells were blocked with PBS containing 2% bovine serum albumin, 2% normal goat serum, and 0.4% Triton-X100. To label HA-tagged PhotonSABER, cells were incubated with rat monoclonal anti-HA antibodies  for 60 min and visualized with Alexa405-conjugated secondary antibodies . Biotinylated EGF was visualized using Alexa554-conjugated streptavidin . In the representative images, constant levels of brightness and contrast were used throughout each experimental series.


Glycyrrhiza contains diverse range of biological compounds including triterpenes  and flavonoids . The flavonoids in licorice are known to have strong anti-cancer activities. Quercetin, the most abundant flavonoid, has been shown to have anti-ulcer, anti-cancer, antioxidant, and anti-inflammatory properties. Latent Epstein-Barr virus (EBV) infection can lead to serious malignancies, such as, Burkitt’s lymphoma, Hodgkin’s disease and gastric carcinoma(GC), and (Epstein-Barr virus associated gastric carcinoma) EBVaGC is one of the most common EBV-associated cancers. In this study, the authors first examined the anti-cancer effects of quercetin and isoliquiritigenin in vivo xenograft animal models implanted with EBV(+) human gastric carcinoma (SNU719) or EBV(−) human gastric carcinoma (MKN74), and then explored the molecular mechanisms responsible for their anti-cancer activities. The results obtained showed that anti-cancer effect of quercetin was greater than isoliquiritigenin in mice injected with EBV(+) human gastric carcinoma (SNU719) cells. On the other hand, quercetin and isoliquiritigenin had similar anti-cancer effects in mice injected with EBV(−) human gastric carcinoma (MKN74) cells. Interestingly, quercetin inhibited EBV viral protein expressions, including EBNA-1 and LMP-2 proteins in tumor tissues from mice injected with EBV(+) human gastric carcinoma. Quercetin more effectively induced p53-dependent apoptosis than isoliquiritigenin in EBV(+) human gastric carcinoma, and this induction was correlated with increased expressions of the cleaved forms of caspase-3, -9, and Parp. In EBV(−)human gastric carcinoma (MKN74), both quercetin and isoliquiritigenin induced the expressions of p53, Bax, and Puma and the cleaved forms of caspase-3 and -9 and Parp at similar levels.Licorice extracts have been widely used in herbal and folk medications.  Glycyrrhiza glabra or Glycyrrhiza uralensis, a perennial legume found in southern Europe (Glycyrrhiza glabra) and in east Asia . Licorice extracts have been widely used in herbal and folk medications. Glycyrrhiza contains a diverse range of biological compounds including triterpenes  and the flavonoids  /2 as previously described [3, animals were euthanized and tumors were harvested.Animal experiments were conducted in accordance with the National Research Council’s Guide  for the Care and Use of Laboratory Animals. The experimental protocol was approved by the Animal Experiments Committee of Duksung Women’s University (permit number: 2014-016-007). NOD/SCID mice  were used as xenograft animal models. Mice were individually accommodated in a pathogen free controlled environment (23–27 °C under a 12-h day/12-h night cycle) and provided food and water ad lib. To produce tumors, mice were first divided into two groups  and EBV(−) human gastric carcinoma SNU719 or MKN74 bearing mice and lysed using buffer solution . Protein level in tumor tissue lysates was measured using the Bradford Assay Plus Kit , separated by electrophoresis, and transferred to nitrocellulose membranes. Membranes were incubated with first and second antibodies and blots were detected using enhanced chemiluminescent (ECL) reagents . The primary antibodies used were anti-p53 , anti-p21, anti-Bax , Apoptosis Sampler Kit , and anti-β-actin Ab (Sigma Aldrich) ,40.p values of less than 0.05 were considered statistically significant. The significances of intergroup difference were determined by ANOVA. In this study, we assessed the anti-cancer effects of quercetin and isoliquiritigenin using in vivo xenograft animals implanted with EBV(+) or EBV(−) human gastric carcinoma. Both quercetin and isoliquiritigenin exhibited anti-cancer effects on EBV(+) and EBV(−) human gastric carcinoma. However quercetin had the greater effect on the EBV(+) gastric carcinoma, which appears to be in line with anti-EBV effect of quercetin.In addition, the protein expressions of p53, p21, Bax, and PUMA were elevated in EBV(+) human gastric carcinoma bearing animals fed quercetin or isoliquiritigenin, but only quercetin did so significantly. Quercetin also increased the expressions of the cleaved forms of caspase-3, -9, and Parp in EBV(+) human gastric carcinoma markedly more than isoliquiritigenin. However, both quercetin and isoliquiritigenin markedly upregulated the cleaved forms of caspase-3, -9, and Parp in EBV(−) human gastric carcinoma (MKN74), which matched their anti-cancer effects in tumor cells without EBV. We conclude that quercetin is a good candidate anti-cancer agent in EBV-associated human gastric carcinoma.


Supramolecular gels containing porphyrins and phthalocyanines motifs are attracting increased interests in a wide range of research areas. Based on the supramolecular gels systems, porphyrin or phthalocyanines can form assemblies with plentiful nanostructures, dynamic, and stimuli-responsive properties. And these π-conjugated molecular building blocks also afford supramolecular gels with many new features, depending on their photochemical and electrochemical characteristics. As one of the most characteristic models, the supramolecular chirality of these soft matters was investigated. Notably, the application of supramolecular gels containing porphyrins and phthalocyanines has been developed in the field of catalysis, molecular sensing, biological imaging, drug delivery and photodynamic therapy. And some photoelectric devices were also fabricated depending on the gelation of porphyrins or phthalocyanines. This paper presents an overview of the progress achieved in this issue along with some perspectives for further advances. Supramolecular gels Steed, , in whicTetrapyrrole macrocycle based π-conjugated systems, such as porphyrin and phthalocyanine derivatives , are verAlthough the advantages of supramolecular gels based on the assembly of porphyrins and phthalocyanines are distinct, building these systems has been demonstrated to be not easy. This situation is partly due to the difficulty of the synthesis of gelators containing porphyrin or phthalocyanine motifs  derivatives.1) (2)n, which connects the porphyrin moiety with the cholesterol groups. After adding fullerene C60 into the systems, the intermolecular Zn(II) porphyrin-fullerene interaction can enhance the gelation abilities of the corresponding porphyrin gelators (2)  , and fougelators  (Ishi-i gelators . Many chgelators . Porphyrtors (2) , have betors (2) . Compounnofibers , wherein60 has attracted a lot of attention. Shinkai et al. have synthesized porphyrin bearing eight amide moieties (3)  . The cooructures . Both po4, 5) substituted by didodecyl L-glutamic acid. These porphyrin molecules can be either derived from protoporphyrin IX (4) or TPP based porphyrin (5)  . These Lyrin (5)  (Sagawa yrin (5) .6) shows much better gelation properties in different organic solvents, compared with that of symmetrical reference porphyrins bearing two pyridyl substituents (8) or two carboxylic acid groups (7)  . In cycl9). The host-guest interactions between pillar[5]arenes and viologen could render the porphyrin to assemble into linear supramolecular polymer with head-to-tail manner  can form organogels in different organic solvents, while the corresponding monomer (11) self-assemble into vesicles in chloroform/methanol mixture  (12 containing (S)-chiral centers could follow a typical hierarchical process. Firstly, the stacking of phthalocyanine 12 along clockwise orientation forms the fibers with right-handed helicity. And then, coiled-coil interactions between different right-handed helical fibers could form large aggregates with left-handed helicity  . This phhelicity  (Engelkahelicity .13) can form organogels in 1,2-dichlorobenzene upon ultrasonication  phthalocyanine (nication  (Xu et anication .14), which could form hydrogels . Copolymer 15 can form hydrogels, which show near infrared fluorescence  (PEG) and ε-caprolactone produces a type of copolymer  form thixotropic and reversible supramolecular hydrogels , which help the amphiphilic porphyrin-cored, star-shaped poly (e-caprolactone)-b-poly (ethylene glycol) (SPPCL-b-PEG) copolymer  and zinc phthalocyanine moieties containing complementary organogelator (20-24). And further in situ cross-linking could enhance the strength of the gels  and N,N'-bis(octadecyl)-D-Boc-glutamic diamide (25-D)  has been included upon mixing without damaging the mechanical strength of the corresponding gels formed in DMSO. Most interestingly, these glutamic acid-based gelators can induce achiral porphyrin form supramolecular chiral assemblies -L-Boc-glutamic diamide (e (25-D) . 25 can Except for glutamic acid-based gels, the hydrogels formed by aromatic short peptide Fluorenylmethoxycarbonyl-Leucine-Leucine-Leucine-OMe (Fmoc-L3-OMe) can also be doped with zinc porphyrin. And enhanced photocurrent generation was investigated from this hydrogel system  Feng et.27) was also found to form gels with hollow fibrous nanostructures, which can encapsulate phthalocyanine molecules (28). And phthalocyanine molecules (28) form crystal within the gels -trans-1,2-bis(dodecanoylamino)cyclohexane. When octakis-substituted Zn(II)-phthalocyanines were incorporated into the gels, unique brush-like nanostructures can be detected  and TPPS (tetrakis(4-sulfonatophenyl)porphyrin) (39) have been doped into the polymer hydrogels fabricated by copolymers of HEMA (2-hydroxyethyl methacrylate) with either MAA (methacrylic acid) or DEAEMA (2-(diethylamino)ethyl methacrylate)  included in the hydrogels can generate singlet oxygen porphyrin) (crylate) . Upon phDoping with different functional molecules is one of the distinctive features of polymer gels. And different polymer gels systems, which can be fabricated by polypyrrole   . The int42TEOS) ,b.43), which can crosslink covalently with chitosan containing amino groups to form dynamic schiff-base linkage , the organic helical nanostructures can be transcripted into helical-silica structures, which nicely inherit the organic morphology .44D or 44L). And the co-assembly of these cationic chiral gelators with dianionic tetrakis(4-sulfonatophenyl)porphyrin (39) could form J-aggregates. Interestingly, the chiral signs of porphyrin assemblies were found to follow the supramolecular chirality of 44 assemblies instead of the molecular chirality of 44  was found to form chiral nanotwists and nanotubes in toluene and DMSO, respectively. While the co-assembly of 45 with achiral porphyrin containing long alkyl chains (26) and C60 can also form gel in DMSO by keeping the tubular nanostructures. Both porphyrin and C60 were included in confined nanotubes. Chirality transfer occurred from the host chiral gel matrixes to guest achiral porphyrin in DMSO. Remarkably, the addition of C60 to the porphyrin/gelator gel could invert and further amplify the induced chirality of the porphyrin due to the formation of donor-acceptor pairs. However, in the case of multicomponent toluene gel, 26 and C60 only dissolve in the liquid phase. And no chirality transfer was observed in the toluene gel   . Interes in mice .15). And the hydrogels formed by 15 can be further included with another dye molecule (rhodamine). These dual fluorescent hydrogels can be used for in vivo near infrared fluorescence imaging   . And the48), which can be released dependent on thermoreversible of hydrogels  and lysine hydrochloride (lys × HCl) can be included with magnesium crown-containing phthalocyanines (ydrogels .47) as gelator, the doxorubicin-loaded hydrogel has been developed as dual fluorescent anti-tumor drug delivery system. Using nude mice bearing luciferase expressed hepatic tumor as models, the whole process from the drug delivery to the tumor therapeutic effects were real time visualized simultaneously after administration at interval from 0 to 18 days  incorporated hydrogel  and multi-walled carbon nanotubes, the drug delivery systems with photo response, fluorescence imaging tracking and photothermal remote controlling properties can be achieved , which has been developed as a mimic of superoxide dismutase (SOD). A cytochrome c assay revealed that the 50 could be maintained even after treatment with simulated gastric and intestinal fluids due to the protection of hydrogels and liposomal aggregations  inhibiting tumor growth. Yuasa et al. have designed encapsulating liposomal drugs by using an alginate hydrogel as carrier. The liposomal drug was composed of manganese porphyrin (egations . The oraegations .The most elaborate drug delivery systems based on hydrogels could be the complex of cowpea chlorotic mottle virus (CCMV) particles and guest-modified hydroxylpropyl cellulose (HPC), which were non-covalently crosslinked through the formation of ternary host-guest complexes with cucurbit[8]uril (CB[8]) . When teWhen photosensitizers were irradiated with light in the presence of oxygen, free radicals or singlet oxygen could be generated. And photodynamic therapy (PDT) can be achieved by using these active substances for killing tumor cells, bacteria or viruses  of different illnesses  was used for constructing supramolecular hydrogels upon host-guest interaction with a-cyclodextrin (a-CD). The corresponding supramolecular hydrogels have good biodegradable and biocompatible properties, which can be used as carrier for doxorubicin (DOX) delivery systems. Considering the generation of singlet oxygen upon photoirradiation on porphyrin, the hydrogels for both sustained-release drug delivery and photodynamic therapy (PDT) can be achieved . Comparing to the solution, the hydrogel containing porphyrin shows enhanced fluorescence intensity with much more singlet oxygen generation upon the same laser irradiation. Moreover, much longer tumor retention can be observed due to the low fluidity of the hydrogel  as well as water soluble porphyrin (TMPyP) (hydrogel .43) with NH2 groups on chitosan through a dynamic covalent Schiff-base linkage, pH sensitive photosensitizer delivery hydrogels for localized cancer therapy can be prepared. The hydrogel can release 43 in the acidic environment of tumors directly by evading the circulation system. And the viability of cancer cells incubated with the hydrogel was decreased significantly at acidic pH after laser irradiation  and tailored nitric oxide photodonor (52). Upon visible light irradiation, both singlet oxygen and nitric oxide can be generated for photodynamic cancer and bacterial therapies  diacrylate (PEGDA) hydrogels containing rare earth-doped nanophosphors (UCNPs) and zinc(II) phthalocyanine. Upon 980 nm irradiation, the multicolor upconversion luminescence of UCNPs not only can trigger the polymerization of PEGDA, but also can excite photosensitizer zinc(II) phthalocyanine to produce singlet oxygen .In another approach, the hydrogel containing poly (ethylene glycol) double acrylates (PEGDA), polyethylene glycol 400 (PEG400), zinc phthalocyanine (ZnPc), and phosphotungstic acid (PTA) was prepared. The polymer hydrogel not only prevents diffusion of the photosensitizer, but also maintains a high zinc phthalocyanine concentration around tumor cells for more effective PDT. And phosphotungstic acid also can help the acceleration of singlet oxygen generation .Some supramolecular assemblies can be used for sensing ions, gas or small organic molecules. For these applications, the interactions between sensors and target molecules could produce some special signals, such as electrical signals or fluorescence  (PEG), 5,10,15,20-tetrakis(4-hydroxyphenyl) porphyrin and hexamethylene diisocyanate . The cortal ions . Moreovetal ions .54) has been prepared into complex devices for real-time and indicator-free detection of aqueous nitric oxide. The polymer hydrogel can be photoluminescence, which is sensing specific to NO. And the changes of photoluminescence can be further transformed into electric signal by the photodetector (Another polymer hydrogel containing porphyrin derivative (detector .−1 cm−2), large concentration range of (2–12 mM) and low detection limit (25 μm). This biosensor based on polymer hydrogel has very fast response time (1 s) and high storage stability  hydrogel (MFH) integrated with reduced grapheme oxide (rGO), vinyl substituted polyaniline (VS-PANI), double-deck lutetium Phthalocyanine (LuPc2), and glucose oxidase (GOx). This complex system can be used for electrochemical detection of glucose with high sensitivity (15.31 μA mMtability .SV = 0.00826 μM−1) at oxygen concentrations below 300 mM and is resistant to photobleaching  and very weak molecular aggregation. This hydrogel exhibits oxygen-responsive phosphorescence -5,10,15,20-tetrakis- porphyrin (56), fluorescein isothiocyanate (57) (56), fluorescent antennae dyes (58) and Förster resonance energy transfer (FRET) donor poly (59) into cationic hydrogel  porphyrin phosphor was used as oxygen indicator . This serescence  and can ate (57) , and theate (57) . In thisate (57) . Neverthin vivo catalyzing strongly highlights the advantages of the gels.Metal porphyrins or phthalocyanines are good catalysts for different reactions. Many related systems, such as P450, have been thoroughly investigated  trimethylammonium chloride]) embedded with anionic [Mn(tetrakis(4-sulfonatophenyl)porphyrin)(OAc)] (39a) . The act39a) .Chen et al. have developed poly(ethylene glycol)-based nanocomposite hydrogels containing Fe-octacarboxylic acid phthalocyanine (FeOCAP)/magnetic attapulgite . This sy2 evolution via photoinjection of electrons from the light-harvesting gel network into the Pt nanoparticles. Upon visible light irradiation, the Pd-porphyrin gels doped with Pt nanoparticles show very high efficiency for hydrogen production in aqueous sodium ascorbate solution  without significant degradation during four runs  moieties in vivo  for  in vivo . The gelBy making gels containing porphyrin molecules, more complex catalytic systems with delicate nanostructures and functions can be achieved. Liu et al. have fabricated antioxidant microgel with both glutathione peroxidase (GPx) and superoxide dismutase (SOD) activities. In this case, the main catalytic components of glutathione peroxidase were put onto the surface of ferritin. The resulting seleno-ferritin (Se-Fn) monomers can self-assemble into nanocomposites that exhibit remarkable GPx activity due to the well-organized multi-GPx catalytic centers. On the other hand, since some metal porphyrin molecules are good superoxide dismutase mimic, the gel system mimicking synergistic dual enzyme can be prepared by crosslinking seleno-ferritin nanocomposites with porphyrin .The supramolecular gels containing porphyrin or phthalocyanine building blocks could provide the porphyrin assemblies or phthalocyanine aggregates with nice nanostructures and tailored molecular packing modes. For example, the tetrapyrrole macrocycle based π-conjugated systems can form either J-aggregation or H-aggregation. Different molecular packing modes could render the assemblies with varied spectral characteristics, special energy transfer or electron transfer properties. Therefore, some unusual photophysical, photochemical or electrochemical properties of these assemblies can be expected.3-OMe). These assemblies show great light response properties. In this case, the long-range order assembly of aromatic peptide can act as photoelectron acceptor and conductor to promote the production of photoelectrons by the light antenna porphyrin molecules. And porphyrin can effectively reduce the resistance, facilitating the transfer of photoelectrons  and aromatic short peptide Fluorenylmethoxycarbonyl-Leucine-Leucine-Leucine-OMe (Fmoc-Llectrons .63) could cross-link polypyrrole to form hydrogels with three-dimensional network. Phthalocyanine favors the formation of polypyrrole nanofibers. And the enhanced interchain charge transport of polypyrrole/63 hydrogels resulted in greatly enhanced conductivity and pseudocapacitance compared with pristine polypyrrole  phthalocyanine, which can form supramolecular gels upon ultrasonic irradiation. The corresponding gels can be further fabricated into field-effect transistor . Due to Fabrication of supramolecular gels containing porphyrins or phthalocyanines molecules represents the efforts to build novel functional soft matters. In this case, functional molecular building blocks could organize into hierarchical nanostructures in a controlled manner. Many interesting features and subsequently novel applications of these materials could be developed depending on ordered nanostructures. Notably, the dynamic and stimuli-responsive nature of supramolecular gels has greatly expanded the application of included π-conjugated systems.In principle, gels especially hydrogels can be considered as matter of life, due to many similarities between their nature and that of the organisms (Li et al., All authors listed have made a substantial, direct and intellectual contribution to the work, and approved it for publication.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Since their discovery in 1970, flavin-containing monooxygenases (FMOs) have been studied as Phase 1 xenobiotic metabolizing enzymes that act on sulfur- and nitrogen-containing small molecules. In 2015, we demonstrated that C. elegans (worm) fmo-2 was not only necessary for hypoxic response and dietary restriction-mediated lifespan extension, but was also sufficient to extend lifespan when overexpressed. Consistenet with a conserved role for FMOs as longevity-promoting enzymes, mouse hepatic Fmo3 transcript is highly upregulated by numerous major lifespan extending interventions. A contrasting series of reports, however, have described mammalian Fmo3-mediated production of trimethylamine N-oxide (TMAO) as a risk factor for atherosclerosis and other major diseases. My thesis research aims to define the regulation and function of worm fmo-2 using genetic and biochemical approaches. My data thus far support the hypothesis that fmo-2 acts on sulfur amino acid pathway intermediates to promote longevity and healthspan.


We evaluated optimal duration of dual antiplatelet therapy (DAPT) after second-generation drug-eluting stent (DES) implantation in acute coronary syndrome (ACS).From pooled analysis of three randomized clinical trials , a total of 2,216 patient with ACS undergoing second-generation DES implantation were selected. Each study randomized patients to a short-duration DAPT arm  or a standard-duration DAPT arm . Two-thirds of patients were male, and their mean age was 63 years. Mean DAPT durations were 164 ±76 and 359 ±68 days, respectively. The primary endpoint was composite of cardiac death, myocardial infarction, stent thrombosis, stroke or major bleeding during the first 12 months after implantation, analyzed according to the intention-to-treat population.Demographic characteristics were balanced between groups. Mean DAPT duration was 164 and 359 days, respectively. Primary endpoint occurred in 22 patients with short-DAPT and 21 patients with standard-DAPT . Landmark analysis after six-months, no significant difference in primary endpoint between short and standard duration DAPT .Short-duration DAPT (≤6 months) demonstrated a similar incidence of net adverse cardiovascular and clinical events at 12 months after second-generation DES in ACS compared with standard duration DAPT (≥12 months).NCT00698607), RESET , IVUS-XPL EXCELLENT (ClinicalTrials.gov,  Dual antiplatelet therapy (DAPT) after drug-eluting stent (DES) implantation has become standard therapy.  The optiBy contrast, recommendations for the optimal duration of DAPT have not changed for patients with acute coronary syndrome (ACS), even with the use of second-generation DESs, as data to support recommendation modifications are lacking. ACS is well known to increase the risk of recurrent ischemic events, and a standard duration of DAPT (≥12 months) may help prevent these events. A recent meta-analysis suggested that the net clinical benefit of different DAPT durations might vary according to the clinical presentation.  However,Data are currently limited regarding differences in clinical outcome between short-duration and standard-duration DAPT after second-generation DES implantation in the ACS population. Therefore, we performed pooled analysis of three randomized trials to investigate the efficacy and safety of short-duration (≤6 months) compared with standard-duration (≥12 months) of DAPT after second-generation DES implantation in patients with ACS.de novo lesion treated with an everolimus or first-generation sirolimus DES. ) and compared using the Student’s t-test. Categorical variables are presented as count (percentage) and compared using χThe consistency of treatment effect in pre-specified subgroups was assessed using Cox regression models with tests for interaction. We applied the DAPT score , PRECISEOf the 2,216 patients, 1,119 were allocated to receive short-duration DAPT and 1,097 to receive standard-duration DAPT. Mean DAPT durations were 164 ±76 and 359 ±68 days for the two groups. Baseline clinical characteristics of the groups were well balanced . Mean agPrimary endpoints occurred in 22 patients with short-duration DAPT group and 21 patients in standard duration DAPT group  and each individual endpoint between two groups . LandmarIn pre-specified subgroup analyses, standard duration of DAPT therapy increased primary endpoint within chronic kidney disease (creatinine clearance ≤ 60mL/min) group compared to short duration of DAPT. there was a significant interaction for the chronic kidney disease and duration of DAPT  at 12 months after second-generation DES implantation between short (≤6 months) and standard-duration DAPT (≥12 months). However, TVR increased with short-duration DAPT compared with standard duration DAPT.Although early DESs reduced in-stent restenosis rate compared with bare metal stents, the concern arose that DESs may have a higher incidence of late and very late stent thrombosis.  Early diHowever, second-generation DESs, with their reduced strut thickness, biocompatible polymer, and appropriate drug dose, has been shown to improve stent strut coverage, reducing stent thrombosis.  In the DSeveral randomized studies compared the safety and efficacy of short-term (≤6 months) versus long-term (≥12 months) DAPT after second-generation DES implantation. The ITALIC trial showed no difference in the primary endpoint  at 12 months between 6-month and 24-month DAPT after everolimus eluting DES implantation.  The TVR ACS itself is a risk factor for future thrombotic cardiovascular events, and 12 months of DAPT has been recommended in practice.  In ACS pOur study demonstrated that short-duration (≤6 months) DAPT was associated with a similar rate of net adverse cardiovascular and clinical events in patients with ACS (including MI) treated with second-generation DESs. Landmark analysis showed no divergence of the curves for primary endpoint after 6 months following stent implantation between short-duration and standard-duration DAPT. The cumulative low incidence of stent thrombosis or MI after second-generation DES implantation might have contributed to these results, suggesting that short-duration (≤6 months) DAPT might be applied for selected patients with low ischemic risk of ACS after second-generation DESs. Although some discrepant results were observed regarding the efficacy and safety of long-term DAPT after DES implantation, recent PCI guidelines indicate that 6-month DAPT is recommended after second-generation DES implantation for patients with ACS and those with a higher bleeding risk.  The currIn a subgroup analysis, significant interaction was shown between chronic kidney disease and duration of DAPT. Chronic kidney disease is well known to be a risk factor for both ischemic and bleeding clinical events.  For net Based on PRECISE-DAPT score, ‘very low’ bleeding risk group showed lower net clinical and cardiovascular events in standard-duration of DAPT compared to short-duration of DAPT in our study. These results might suggest longer DAPT for low bleeding risk with high ischemic risk group such as ACS. But, overall borderline interaction with the PRECISE-DAPT score indicated that a personalized approach might be the best option to optimize a clinical outcome.Current guideline recommended to decide duration of DAPT based on bleeding risk score system that is not enough to say ‘personalized DAPT’.  To develCoronary artery disease is complex and broad-spectrum disease that many factors should be taken into consideration. However, prediction model based on traditional mathematical model is too complex to apply in clinical practice. Artificial neural networks and sustainable machine learning systems might be helpful in solving dilemmas of optimal DAPT duration raised by our meta-analysis.  However,This study has limitations. First, sample size of the study population might be not sufficient to evaluate the safety of short-duration DAPT compared to longer-duration DAPT in terms of hard clinical endpoints such as death, MI, or stent thrombosis. Second, all 3 trials were open-label and not placebo-controlled studies; however, the possibility of bias was minimized in each trial by using precisely-defined criteria for the primary endpoint, blinding the adjudication by event-adjudication committee members, and analyzing the data using intention-to-treat methodology. Third, short-duration DAPT was 3 months (RESET) or 6 months (Excellent and XPL-IVUS), and the Endeavor zotarolimus-eluting stent was included in the 3-month DAPT group in the RESET study. Fourth, clopidogrel was used with aspirin as DAPT in all 3 trials, so we could not evaluate the effects of DAPT duration for other antiplatelet agents, such as ticagrelor or prasugrel. Fifth, 1 year of clinical follow-up may not be sufficient to assess late outcomes, especially very late stent thrombosis. Nevertheless, only a few existing clinical studies have been performed with adequate number of populations for optimal duration of DAPT in ACS patients. In this perspective, our pooled patient-level analysis may provide an insight to how to apply an optimal duration of DAPT in current DES era with reasonable number of patient population with ACS.In patient-level pooled analysis, short-duration (≤6 months) DAPT showed no difference in incidence of net adverse cardiovascular and clinical events, including cardiac death, MI, stent thrombosis, stroke or major bleeding at 12 months after second-generation DES implantation in patients with ACS compared with standard-duration (≥12 months) DAPT. Composite of cardiac death, MI or stent thrombosis or each component also did not differ according to DAPT duration. These findings should be confirmed with large randomized clinical trials.S1 Fig(TIF)Click here for additional data file.S1 Table(PDF)Click here for additional data file.S2 Table(PDF)Click here for additional data file.S1 File(PDF)Click here for additional data file.S2 File(PDF)Click here for additional data file.S3 File(PDF)Click here for additional data file.S4 File(CSV)Click here for additional data file.


BRAF (B-Raf proto-oncogene) mutations than in those with RTK (receptor tyrosine kinase) or RAS (rat sarcoma) mutations. Hence, inhibition of the mitogen-activated protein kinase kinase-1 and -2 (MEK-1 and -2) downstream of RAF (rapidly accelerated fibrosarcoma) could sensitize RAI refractivity in thyroid cancer. However, a significant hurdle is the development of secondary tumor resistance (escape mechanisms) to these drugs through upregulation of tyrosine kinase receptors or another alternative signaling pathway. The sodium iodide symporter (NIS) is a plasma membrane glycoprotein, a member of solute carrier family 5A (SLC5A5), located on the basolateral surfaces of the thyroid follicular epithelial cells, which mediates active iodide transport into thyroid follicular cells. The mechanisms responsible for NIS loss of function in RAI-R thyroid cancer remains unclear. In a study of patients with recurrent thyroid cancer, expression levels of specific ribosomal machinery—namely PIGU (phosphatidylinositol glycan anchor biosynthesis class U), a subunit of the GPI  remains a mainstay of therapy for these patients. Unfortunately, many thyroid cancer patients have tumors that no longer trap iodine, and hence are refractory to RAI, heralding a poor prognosis. RAI-refractory (RAI-R) cancer cells result from the loss of thyroid differentiation features, such as iodide uptake and organification. This loss of differentiation features correlates with the degree of mitogen-activated protein kinase (MAPK) activation, which is higher in tumors with  Radioactive iodine I-131 (RAI) is a cornerstone in the routine adjuvant management in patients with high-risk differentiated thyroid cancer (DTC) ,3,4. PatWith recent advances and developments in understanding of the oncogenic pathways involved in the development of thyroid cancers and the molecular basis of RAI refractoriness, targeted therapies are being developed and are showing promising results ,8,9,10. i.Absence of RAI uptake at initial diagnosis of locoregional recurrence or distant metastasis;ii.Absence or progressive loss of radioiodine uptake in the post-therapy scan several days after RAI therapy;iii.Presence of more than 1 metastatic lesion with at least one lesion without RAI uptake in the post-therapy scan;iv.Structural progression of tumors 12 to 16 months after RAI therapy despite the presence of iodine uptake in the post-therapy scan;v.Tumors in patients who have received 600 millicurie (mCi)/22.2 gigabecquerel (GBq) or more of RAI cumulatively without signs of remission;  viSignificant uptake on 2-deoxy-2-[fluorine-18] fluoro-D-glucose positron emission tomography integrated with computed tomography (F-18 FDG PET/CT).A major obstacle to standardizing the approach to RAI-R tumors is the lack of a consistent definition for RAI-R. The following definitions for RAI-R tumors are used in the literature ,12,13:i.None of the above criterion alone portends that a tumor is RAI-R, rather it predicts the likelihood that a tumor will be RAI-refractory and should be used together for risk stratification of the tumors in assessing for refractoriness. It is important for the RAI uptake scan to be standardized and an optimal approach is required for patient preparation and choice of imaging modality .BRAF and telomerase reverse transcriptase (TERT) promoter mutation) can also be included for risk stratification of patients [Aggressive or poorly differentiated tumors on histology or tumors exhibiting aggressive genetic profiles (such as patients .SLC5A5), located on the basolateral surface of the thyroid follicular epithelial cells. NIS mediates active iodide transport into thyroid follicular cells, and in the normal thyroid cells, Thyroid stimulating hormone (TSH) stimulates NIS expression via the cyclic adenosine monophosphate (cAMP) pathway, binding to the NIS promoter via paired box 8 (PAX8) [RAI refractoriness most frequently develops in the context of loss of thyroid differentiation features. One hallmark of dedifferentiation is impairment of Na/I symporter (NIS) function ,17. NIS 8 (PAX8) . The abi8 (PAX8) ,19. In a8 (PAX8) . Figure 8 (PAX8) .V600EBRAF mutation. V600EBRAF represents the most frequent genetic aberration in thyroid cancers, occurring in nearly 50% of DTCs. V600EBRAF mutations both inversely correlate with NIS expression and directly correlate with dedifferentiation, recurrence, and metastasis [NIS promoter in follicular cells [NIS by driving histone deacetylation of the H3 and H4 lysine residues of the NIS promoter, directly preventing its transcription [Genetic and epigenetic alterations in the RTK/BRAF/MAPK/ERK and PI3K-AKT-mTOR pathways by acquired point mutations, chromosomal rearrangement, or aberrant gene methylation underly the diminished NIS signaling central to RAI refractoriness ,22,23. Ttastasis ,25. BRAFar cells . Second,cription .RAI-R metastatic DTCs can be asymptomatic for several years . Active In locoregional relapse, surgery is still the most commonly used therapeutic approach, with therapeutic compartmental central or lateral neck dissection to spare uninvolved vital structures, or a more limited surgery in cases of prior comprehensive neck dissection ,13. ExteTargeted treatments for thyroid cancer have been increasingly developed over the last decade along with increasing knowledge about the disease’s underlying molecular alterations. Most agents that were tested in phase II and III trials were developed for treatment of advanced RAI-R thyroid cancer. Cellular dedifferentiation in thyroid cancers causes tumor progression in the form of more aggressive growth, metastasis, loss of iodide uptake, or unresponsiveness to RAI therapy, and correlates with the degree of MAPK activation. Tyrosine kinases are involved in the MAPK signaling pathway through phosphorylation/dephosphorylation of several intracellular proteins, which underlies the rationale for the use of tyrosine kinase inhibitors (TKIs) in the treatment of thyroid cancer ,39.TKIs have been shown to significantly improve progression-free survival rates in advanced RAI-R DTCs. Overall survival has been difficult to document in these trials, likely because patients cross over to the drug arm once disease progression is documented in the control group ,10. An oAlthough TKIs have revolutionized the field of targeted therapy in RAI-R DTC patients, these agents are usually administered lifelong, and several drawbacks were associated with their long-term application. TKIs’ adverse effect profiles have a great impact on quality of life and should be taken into consideration by the clinician prior to treatment initiation. Also, a resistance to the treatment, also called “escape”, can develop; hence, access and adherence to close monitoring with a continuous assessment of adverse effects and the patient’s quality of life should be considered in the decision to start the therapy . In clinPotential targets are also transcriptional factors such as vascular endothelial growth factor (VEGF). In response to intratumoral hypoxia, hypoxia inducible factor-1 alpha (HIF-1α) is activated and induces VEGF transcription together with co-stimulation by growth factor signaling pathways, such as the PI3K/AKT and MAPK pathways ,47. VEGFAs discussed above, thyroid cancers often become RAI-R by co-opting RAF and RAS signaling, thereby repressing NIS and RAI uptake. Although targeted therapies such as BRAF inhibitors have shown some success in resensitizing tumors to RAI, these tumors often escape RAI sensitivity via aberrations in complementary pathways. In patients with documented response to targeted therapies, after several months, the tumor escapes . Several hypotheses and suggested mechanisms have been proposed to explain such escape, most of which involve overactivation of alternative pathways to overcome the drug’s effect ,51.Combining adjuvant therapies with TKIs has the potential to eliminate or delay the escape effect  and result in longer progression-free survival. Numerous genetic and signal transduction alterations have been observed in RAI-R or advanced papillary thyroid cancer (PTC) ,21,23, aBRAF mutant cells treated with the BRAF inhibitor vemurafenib [One unique mechanism by which thyroid cancer drives RAI resistance may be via upregulation of the human epidermal receptor (HER) family of receptor tyrosine kinases, which is present in more than one-third of thyroid cancers and is positively correlated with local tumor invasiveness . HER2 anurafenib . Therefourafenib .EML4) gene—that were described in a subset of patients, who tend to be younger with more aggressive disease [Anaplastic lymphoma kinase (ALK) is a recently identified kinase with the potential to contribute to aggressive disease in non-small-cell lung cancers. This kinase can undergo rearrangements—the most common is EML4-ALK fusion with echinoderm microtubule–associated protein-like 4  and PTTG-1 binding factor overexpression in thyroid cancers results in decreased NIS levels , likely Phase II trials of the efficacy of various chemotherapy agents for recurrent and metastatic DTC have been reported; in these, doxorubicin was the most frequently used agent . No consBRAF-V600E-positive DTC revealed high levels of PD-L1 (Programmed death-ligand 1) (53% vs. 12.5%) and human leukocyte antigen G (41% vs. 12.5%) compared with BRAF wild-type tumors. Furthermore, BRAF-V600E-positive tumors had a high level of suppressive T cell and macrophage components [Characterizing the immune landscape following TKI treatment in various tumor types, including thyroid cancer, has demonstrated dynamic alterations in the tumor immune microenvironment ,74. UsinOngoing trials are evaluating the role of TKIs in advanced RAI-R thyroid cancer, either alone or in combination with immune checkpoint inhibitors or RAI therapy . SulfatiCurrent American Thyroid Association (ATA) guidelines recommend high-risk metastatic progressive  RAI-R-DTCs not amenable to conventional therapies be considered for TKIs in specialized centers. Since immunotherapies and re-sensitization therapies are currently in the phase of clinical trials, the ATA recommends admittance into these trials if RAI-R DTCs are progressive after use of approved TKIs, such as lenvatinib or sorafenib. Molecular characterization of these lesions can help to identify and select the appropriate clinical trials. The ATA endorses the use of EBRT or radiofrequency ablation or cryoablation over surgery for symptomatic distant metastatic lesions or lesions with high risk of local complications prior to initiation of TKIs. It also advocates their use for single or multiple progressive lesions while on TKIs or other novel therapies . BRAF-V600E-positive RAI-R PTCs [Novel therapies with single kinase inhibitors have been shown to re-induce iodide uptake in RAI-R thyroid cancer cells. The MEK 1/2 inhibitor selumetinib was found to reverse refractoriness to RAI in patients with advanced or metastatic DTC . RothenbI-R PTCs . Sabra eI-R PTCs . A majorI-R PTCs ,80.BRAF-V600E-positive thyroid cancer cells to RAF or MAP/ERK inhibitors [BRAF-V600E-positive PTC to a BRAF/MEK inhibitor by preventing MAPK rebound and increased NIS expression [BRAF mutated RAI-R DTC [BRAF inhibitors  and MEK inhibitors  activate PI3K and MAP-K pathways by inducing HER3 gene expression. The HER3 inhibitor lapatinib prevents this MAPK rebound and sensitizes hibitors . Use of pression . These fAI-R DTC .BRAF-mutated thyroid cancer, resulting in restoration of RAI uptake through increased NIS expression [RAS and BRAFV600E mutations [ERK rebound could be inactivated in a sustained manner with the use of a MEK inhibitor and by thwarting RAF reactivation in pression . This coutations .Efficacy and toxicity should be considered in the choice of agents to treat RAI-R thyroid cancer. These agents include axitinib, cabozantinib, pazopanib, sorafenib, sunitinib, and vandetanib, including the selective BRAF inhibitors vemurafenib and dabrafenib . RAF and MEK inhibitors have shown promise in the redifferentiation of RAI-R DTC; verification of these results could improve treatment options for patients with RAI-R DTC. New targeted agents, immune checkpoint inhibitors, and combinations of agents for redifferentiation show promise and could improve the efficacy of RAI therapy for patients with RAI-R thyroid cancer.


Head and neck squamous cell carcinoma (HNSCC) represents a model of escape from anti-tumor immunity. The high frequency of p53 tumor suppressor loss in HNSCC leads to genomic instability and immune stimulation through the generation of neoantigens. However, the aggressive nature of HNSCC tumors and significant rates of resistance to conventional therapies highlights the ability of HNSCC to evade this immune response. Advances in understanding the role of co-stimulatory and immune checkpoint receptors in HNSCC-mediated immunosuppression lay the foundation for development of novel therapeutic approaches. This article provides an overview of these co-stimulatory and immune checkpoint pathways, as well as a review of preclinical and clinical evidence supporting the modulation of these pathways in HNSCC. Finally, the synergistic potential of combining these approaches is discussed, along with an update of current clinical trials evaluating combinations of immune-based therapies in HNSCC patients. Head and neck squamous cell carcinoma (HNSCC) is the sixth most common cancer in the world, affecting over 500,000 people each year . While HIn a process termed immune surveillance, recognition of non-self antigens on tumor cells allows for their destruction by the host immune system . The higOnce recruited to the tumor microenvironment, T-cells interact with antigen-presenting cells (APCs) at the “immune synapse,” and require two simultaneous signals from APCs before they can be activated to mediate their anti-tumor effects Fig. . The firIn contrast to these co-stimulatory molecules, the inhibitory “immune checkpoints” prevent T-cell activation. These checkpoints normally function to prevent exaggerated immune responses and subsequent autoimmune disease. However, HNSCC subverts this physiologic function in order to suppress tumor-directed immune activation. The two best known checkpoints include programmed death-1 (PD-1) and cytotoxic T-lymphocyte-associated protein 4 (CTLA-4), both of which are the targets of FDA-approved inhibitory antibodies .Like other cancers, HNSCC tumor cells mediate immunosuppression in the tumor microenvironment (TME) through mechanisms including upregulation of PD-L1 expression and release of immunosuppressive factors . In addiAs mentioned previously, the co-stimulatory receptors that make up “signal two” play a central role in the activation of tumor-fighting T-cells. Absence of this co-stimulatory signal can lead to induction of T-cell anergy or apoptosis and decreased strength of the immune response. In addition to the classically described CD28/B7, other key co-stimulatory interactions between T-cells and APCs, respectively, include CD137/CD137-L, OX40/OX40-L, and CD40-L/CD40  is a costimulatory receptor expressed on the surface of activated T-cells, NK cells, and dendritic cells. When bound by its ligand (CD137-L) on the surface of APCs such as macrophages, dendritic cells, and B cells, trimerization of activated CD137 enhances proliferation, cytotoxic capacity, and survival of T-cells . StimulaTwo humanized monoclonal antibodies (mAb) against CD137 have been developed including urelumab (IgG4) and PF-05082566 (IgG2) . These aCD40-L expressed on the surface of activated CD4 T-cells binds to CD40 on APCs, playing a key role in the “helper” T-cell function to activate APCs to prime CD8 T-cells . ExpressA variety of CD40-targeting therapies have been developed, including agonistic mAbs and recombinant ligands. Although this approach has not been studied specifically in the HNSCC population, one HNSCC patient treated in a phase I trial of recombinant CD40-L experienced a durable and complete response . Phase ILike CD137, OX40 is a co-stimulatory molecule expressed on the T-cell surface that promotes T-cell proliferation, cytokine secretion and memory function when bound by its ligand (OX40-L) . The relA phase I trial of the murine agonistic anti-human OX40 mAb 9B12 in patients with refractory solid tumors demonstrated a mild toxicity profile and promising immunologic correlates (NCT01644968) . AlthougA reciprocal approach to agonism of co-stimulatory receptors is the inhibition of the immunosuppressive checkpoint receptors. These molecules are upregulated by immune activation, and serve a physiologic role by preventing excessive inflammation and autoimmune disease. However, when overexpressed in the TME, these checkpoints contribute to tumor-promoting immunosuppression, and therefore represent a promising target for disinhibiting immune responses against tumor cells and improving HNSCC patient outcomes. Immune-related adverse events (irAEs) may occur in patients treated with these drugs, including rash, gastrointestinal symptoms, thyroid disorders or autoimmune pneumonitis; however these irAEs are now easily recognized and treated with steroids and/or cessation of the drug in most cases .PD-1 is expressed by activated CD8 T-cells, NK cells, B cells, monocytes, and dendritic cells, and normally serves to prevent overactivation of the immune response . HoweverA variety of antibodies have been developed against both PD-1 and PD-L1, including pembrolizumab  and nivolumab . Numerous clinical trials targeting the PD-1/PD-L1 pathway have been extensively discussed in recent reviews , 13, 43.CTLA-4 is transiently expressed by activated T-cells upon binding of an antigen-bearing MHC molecule to the TCR, thereby limiting exaggerated immune responses . CTLA-4 Since that time, two humanized anti-CTLA-4 mAbs, ipilimumab (IgG1) and tremelimumab (IgG2), have been developed and evaluated in phase III trials in advanced melanoma . Based oLAG-3 in murine models has been shown to reduce the immunosuppressive activity of Tregs, and conversely ectopic expression of LAG-3 has been shown to confer immunosuppressive capacity upon CD4 T-cells [LAG-3 is another inhibitory checkpoint that is expressed on the surface of Tregs in HNSCC patients . LAG-3 h T-cells . In addi T-cells . Anti-LA T-cells . In muri T-cells . Early pTIM-3 represents an additional inhibitory checkpoint that has been implicated both in the immunosuppressive function of Tregs and in the exhaustion of effector T cells in the TME. Elevated expression of TIM-3 has been observed on intratumoral Tregs derived from patients with HNSCC and non-small cell lung cancer, and has been observed to correlate with worse clinical outcomes , 59. TIMInitially identified as a co-stimulatory receptor of T cell function , B7-H3 hAlthough many of the above mentioned immunotherapy approaches have shown significant efficacy in certain patients, there is room for improvement with regards to expanding response rates. Current efforts focus on the rational combination of immunotherapy approaches in order to increase the breadth and depth of patient responses Table . It is iCetuximab is a human-mouse chimeric IgG1 antibody against epidermal growth factor receptor (EGFR) that is FDA approved as a monotherapy for recurrent/metastatic HNSCC, in combination with radiation therapy for advanced HNSCC, and in combination with chemoradiation for recurrent/metastatic HNSCC . AlthougIn addition to mediating ADCC, cetuximab-activated NK cells have been shown to promote maturation of APCs and the development of an adaptive immune response . Recent Human NK cells have been shown to upregulate surface expression of CD137 following exposure to cetuximab and EGFR-expressing cell lines . In precEx vivo treatment of HNSCC tumor-infiltrating lymphocytes with the anti-CTLA-4 mAb ipilimumab depleted Tregs and restored NK cell-mediated ADCC [In addition to FcγRIII polymorphisms, proposed mechanisms of resistance to cetuximab-mediated ADCC include an increase in the number of Tregs within the HNSCC TME following cetuximab treatment . These Cted ADCC . Based oToll-like receptors (TLRs) are transmembrane receptors that recognize microbial invasion and respond through activation of the innate immune system . TLR7 anBased on these promising preclinical data regarding selective TLR8 agonism, VTX-2337 was studied in a phase I trial in advanced solid tumors . This trAlthough the degree and durability of response to checkpoint inhibitor monotherapy has been impressive, objective response rates remain low. For example, preliminary data from the KEYNOTE-012 expansion cohort showed an objective response in 18.2 % of recurrent/metastatic HNSCC patients treated with pembrolizumab monotherapy . For thiSignaling through various immune checkpoints and downregulation of costimulatory receptors each represent a distinct mechanism of tumor-mediated immunosuppression. Combining inhibitors that target different checkpoints is a logical strategy to generate synergy and target potential mechanisms of resistance to therapy. For example, melanoma patients with high PD-L1 expression did not respond to anti-CTLA4 mAb and radiation, implicating PD-1/PD-L1 signaling in this resistance (NCT01497808) . HoweverA phase I trial of combined nivolumab anti-PD-1) and ipilimumab (anti-CTLA-4) in advanced melanoma showed an overall response of 40 % and objective responses in 53 % of patients treated with the maximal tolerated dose . These o and ipilIn addition, simultaneous agonism of co-stimulatory pathways and antagonism of inhibitory checkpoints allows one to “step on the gas while taking the foot off the brakes.” In solid tumor models, combined treatment with agonistic anti-OX40 mAb and anti-CTLA-4 mAb improved survival and induced tumor regression through expansion of effector CD8 T-cells . CombineIn addition to combination immunotherapies for HNSCC, other promising strategies under investigation include the combination of these agents with standard-of-care or targeted therapies. Studies in HNSCC and other cancer types suggest that radiotherapy, cisplatin chemotherapy, and other cytotoxic drugs may enhance anti-tumor adaptive immunity in the TME –90. As aThe Cancer Genome Atlas and other studies have revealed specific genomic alterations in HNSCC that may be targeted by specific therapies . AlthougAs mentioned above, HPV-associated HNSCC represents distinct mechanisms of antiviral immune escape, and this disease entity also may benefit from specific antiviral immunotherapies. Since patients with HPV-associated disease generally have an excellent prognosis and high cure rates, newer therapeutic strategies have focused on improving upon long-term toxicities seen with current therapies. Therapeutic vaccines and adoptive transfer of immune cells have been studied in HPV-associated HNSCC  and willRecent advances in understanding the balance between costimulatory and inhibitory immune pathways at the immune synapse have encouraged interest in re-directing these signals from tumor-promoting immunosuppression towards tumor-fighting immunity. With the development of an array of costimulatory agonists and checkpoint inhibitors, these immune-based strategies have become a focal point for research in many cancers, including HNSCC. In addition to focusing on clinical application of these novel immunotherapies, much work is underway to investigate mechanisms of resistance in those patients who do not achieve durable responses. Rational design of combination strategies represents a promising approach to target resistance, while care must be taken to avoid immune over-activation and serious autoimmune consequences.