Graphene and its derivatives have lately been the subject of increased attention for different environmental applications of membrane technology such as water treatment and air filtration, exploiting their antimicrobial and antiviral activity. They are interesting candidates as membrane materials for their outstanding mechanical and chemical stability and for their thin two-dimensional (2D) nanostructure with potential pore engineering for advanced separation. All these applications have evolved and diversified from discovery to today, and now graphene and graphene derivatives also offer fascinating opportunities for the fight against infective diseases such as COVID-19 thanks to their antimicrobial and antiviral properties. This paper presents an overview of graphene-based 2D materials, their preparation and use as membrane material for applications in water treatment and in respiratory protection devices. The COVID-19 pandemic was the defining global health crisis of our time—a novel infectious disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) . This tyInfectious diseases caused by viruses threaten the health of both humans and animals, but often, viral diseases can be alleviated or even eradicated by vaccination . In the Graphene (G) is a carbon sheet one atom thick and made up of condensed six membered rings. It is a two-dimensional allotrope of carbon, separated from graphite for the first time in 2004 by A. Geim and K. Novoselov who were awarded the 2010 Nobel Prize in physics for this discovery . Single-Graphene-based materials are widely applied in green energy such as fuel cell, solar cell, lithium-ion batteries (LIBs), supercapacitor, dye-sensitized solar cells (DSSC), photocatalysis and photo-electrochemical cells for hydrogen evolution via water splitting . ReducedGraphene oxide and its derivatives, such as others nanoscale materials, have been widely explored for their antimicrobial properties due to their high surface-to-volume ratios and unique chemical and physical properties ,25. On tThis review, after an initial overview on application of graphene-based membrane for water purification, focuses on the antibacterial and antiviral activity of graphene-based materials and the possibility of using graphene-based membrane for reusable and recyclable masks.Graphite is a 3D material that is organized and built up by millions of graphene layers. Graphene (G), being one-atom thick layer of extended sp2 carbons, represents the limit of the thinnest possible 2D conductive surface, exhibiting very fast electron mobility and high charge carrier density . Until nOther derivatives of graphite are graphene oxide (GO), reduced graphene oxide (rGO), N-doped G (NG), graphene quantum dots (GQDs) etc. . One of 4 is used for graphite oxidation in the presence of H2SO4, NaNO3, and H2O2. The second is the Brodie method that is performed by adding potassium chlorate to slurry of graphite in fuming nitric acid ..65]. The membranes present a composite structure with 2D layered GO supported on macroporus PAI C,D. The Moreover, new PVDF-GO mixed matrix membranes were also developed using DMSO as non-hazardous solvent, in place of traditional substances of very high concern (SVHC), in a combined vapour- and liquid-induced phase separation process in which water in vapour or liquid form works as a non-solvent for the polymer inducing its precipitation. The protocol developed did not include any chemical additive generally used as pore former  in phase separation process for the production of porous membranes . The preContamination of water with bacteria, protozoans and viruses is a serious problem because water acts as a carrier for infectious pathogens that spread harmful diseases . The literature available regarding the separation of viruses from the water mostly denotes the use of polymeric membranes depending on their pore size . In factQu et al. reported that COVID-19 can survive in stool samples for 4 days and that this virus has been positively detected in stool samples of infected patients . In otheMembranes for water purification and water desalination are being used more and more to address global challenges of pollution and scarcity of water . Highly E. coli and creates strong interactions between itself and lipid molecules in bacteria [Some authors reported that the antibacterial ability of graphene can be increased when doped with metal nanoparticles , becausebacteria . Escherichia coli  and Staphylococcus aureus  [E. coli using recycled graphene oxide–iron oxide–silver nanocomposites [Magnetic graphene-based nanocomposites are used as an antibacterial agent for water purification ,86. Bothositive) . These cositive) . The larmposites . Other sThe growing antibacterial resistance spectrum of bacterial infections required the development of novel and efficient antibacterial agents. Biocidal nanomaterial is usually used in the science field to improve medical devices’ wastewater treatment, food packaging, synthetic textiles and dentistry . The increased attraction towards graphene and graphene derivatives such as graphene oxide and reduced graphene oxide is determined by the bactericidal activity of G on several Gram-negative and -positive bacteria . G is onEscherichia coli investigating its antimicrobial mechanism [Escherichia coli than against Staphylococcus aureus because of the absence of any additional outer membrane in the case of S. aureus. E. coli was more damaged by the GO coatings than S. aureus, probably for the oxidative stress from GO [Sometimes authors reported different behavior of graphene derivatives in terms of antibacterial effect. Liu et al. compared the antibacterial activity of graphite (G), graphite oxide (GtO), graphene oxide (GO), and reduced graphene oxide (rGO) toward a bacterial model such as echanism . The autechanism . Other sechanism . De Fariechanism . These oechanism . Many auechanism ,99. The echanism . For exa from GO . Investigating the interactions of GO with Gram-positive and Gram-negative bacteria, often the antimicrobial effects of GO are concentration and time dependent. The main structural difference between these two types of bacteria is that Gram positive bacteria have a thick peptidoglycan layer and no outer lipid membrane, while Gram negative bacteria have a thin peptidoglycan layer and have an outer lipid membrane. E. coli strain and human HeLa cells. The typical bacterium Escherichia coli and HeLa cell were used to evaluate the antibacterial activity and cytotoxicity of the ZnO/GO composites, respectively. The antibacterial activity of the ZnO/GO composites is dependent on the content of zinc in the composites. It is mainly due to the synergistic effect of ZnO and GO: the possibility of bacteria contact to zinc was increased as ZnO NPs and released zinc ions from ZnO NPs were enriched on the GO sheets. The intimate contact of the E. coli cells and ZnO NPs on the GO sheets enhanced the permeability of the bacterial membrane and the local free zinc concentration around bacteria. The cell viability loss induced by the composites is dose-related. GO exhibited no viability loss, even at relatively high concentrations (50 μg mL−1), indicating the low toxicity of GO to HeLa cells. Similarly, even when the concentration of ZnO/GO-1 (prepared with mass ratio of ZnO/GO was 3:1) reaches 20 μg mL−1, the cell viability is still over 90%. However, a concentration of ZnO NPs (240 μM) decreased the cell viability to 30%. These results illustrate that the ZnO/GO composites cause much lower cytotoxicity than an equivalent amount of ZnO NPs alone. In another study [E. coli after 12 h. This antibacterial activity was probably caused by the synergy of the physical interactions of graphene with bacterial membranes and the antibacterial properties of ZnO that have the aptitude to photo catalytically generate H2O2 and to penetrate cells and disrupt the bacterial membrane. Furthermore, graphene oxide can be easily modified with silver NPs, obtaining the advantages of the physical properties of GO and the strong antibacterial activity of Ag NPs [P. aeruginosa. De Faria et al. [P. aeruginosa over the concentration range investigated. While the GO–Ag nanocomposite showed an antibacterial activity with a minimum inhibitory concentration ranging from 2.5 to 5.0 μg/mL. Moreover, the results showed a 100% inhibition rate of the adhered P. aeruginosa cells on stainless steel surfaces after exposure to the GO–Ag nanocomposite for one hour. This interesting result showed the possibility of using GO–Ag nanocomposites as antibacterial coatings material to avoid the growth of biofilms in food packaging and medical devices.Wang et al.  studied er study , ZnO/graf Ag NPs . Some rea et al.  reportedLukowiak et al.  reportedThe interaction of G-based nanomaterials with bacterial cells depends on their sheet size, surface area, roughness, hydrophilicity, dispersibility and functionalization . The sepThe interaction of bacteria with GO follows a different mechanism when it is in suspension form or immobilized on a coated surface . The phyThe biocidal strength order of Gr-based nanomaterials dispersions is reported as follows: G hybrid NPs, GO and G B; while Despite virus vaccination being the most widely used method for alleviating or eradicating some diseases, attenuated vaccines are not available for all viral infections . The risIn recent years, graphene oxide derivatives were studied to inhibit some viruses such as herpes simplex virus type-1 (HSV-1) competing with the heparan sulfate in binding HSV-1 . One of the major viral pathogens of the lower respiratory tract of infants is the respiratory syncytial virus (RSV). Yang et al.  demonstrVery few studies have reported on the inhibition of the entry and replication of enveloped DNA virus (herpesvirus) and RNA virus (coronavirus) in their target cells by using graphene-based material . Ye et aYe et al. analyzed the impact of GO charge by observing the antiviral activity of GO conjugated with cationic polymer PDDA or nonionic PVP . The resChen et al.  investigA significant advantage of GO-Ag nanocomposites over free Ag NPs is the increasing biocompatibility of the material and the toxicological effects of reducing associated with metallic nanoparticles due to their immobilization on GO. GO-Ag nanocomposites have a specific surface area, are not corrosive and are highly dispersible in water. An important use of GO sheets is as a supporting and stabilizing agent in preventing the agglomeration of the Ag NPs that showed a spherical-like morphology and an average size of 7.5 nm, which is suitable for antiviral activity ,113,118.The study of graphene-based membrane with antiviral activity is still at a nascent stage. Akhavan et al., fabricated Graphene−tungsten oxide composite thin films with sheetlike surface morphology that were applied in photoinactivation of viruses under visible light irradiation . The virGraphene possesses excellent light-to-heat conversion ability owing to its superior thermal conductivity. Deokar et al. designed a graphene-based photothermal antiviral agent that captures and destroys viruses . They syThe main transmission mode of virus such as COVID-19 is through short-range aerosols and droplets . Micron-For this reason, the use of disposable surgical masks or respirator are common for patients, doctors, and even the people in high-risk areas. The United States Centers for Disease Control and Prevention (CDC) recommends the usage of N95 filtering facepiece respirators (FFR) as personal protective equipment for healthcare professionals . The CDCThe N95 FFR is composed of multiple layers of polypropylene with a meltblown layer that is 100−1000 μm in thickness with polypropylene microfibers’ diameters in the range of ∼1−10 μm . These fThe use of disposable surgical masks is common for patients, doctors, and for all people in highly risky areas . As explIn general, the mechanisms of disinfection or sterilization of bacteria and viruses include protein denaturation , DNA/RNA disruption , and cellular disruption , but these methods can modify the masks’ filtration efficiency . Liao et2 laser scribing  filter-based 3D-printed facial protective equipment [Huang et al. identified the bacteria viability on common face masks and found that the majority of bacteria (90%) remain alive after 8 h . The autquipment . The autquipment . Indeed,quipment . For thiquipment  who carrSeveral efforts have been made by the research communities around the globe to contain and reduce the impact of COVID-19. However, the rapid spread of COVID-19 virus was not easily to reduce, and it remains very important to prevent airborne transmission with a respirator filter obtained by using innovative or existing technologies. Very interesting materials that can be used for this purpose are graphene and graphene oxide. This review, after an initial overview on application of graphene-based membrane for water purification, focuses on the antibacterial and antiviral activity of graphene-based materials and the possibility of using graphene-based membrane for reusable and recyclable masks in order to help researchers to find in graphene materials help to fight dangerous viruses and bacteria.The increased attraction towards Graphene and graphene derivatives such as graphene oxide and reduced graphene oxide is determined by the bactericidal activity of G on several Gram-negative and -positive bacteria. G is one of the most recently developed biocidal nanomaterials for its robustness, mechanical strength, great surface area and resistance to degradation, due to its superior surface-to-volume ratio. The attachment and adherence of G to biological cells is due to their active segments, edges, and surfaces. The interaction of G-based nanomaterials with bacterial cells depends on their sheet size, surface area, roughness, hydrophilicity, dispersibility and functionalization.Very recently the possibility was reported to make reusable masks coated on graphene that can be sterilized under sunlight irradiation where the surface temperature can quickly increase to over 80 °C. Other authors produced a prototype for a customizable/multi-used protective respirator mask in which CuO and GO nanoparticles should inactivate viral particles.The study of graphene-based membrane with antiviral activity is still at a nascent stage that needs further work. Indeed, despite the great interest in the use of graphene to modify the mask filters due to its potential use as antibacterial and virucidal properties, additional research efforts should be direct also to the use of new methodologies to complement the actual standard tests and ensure the security of the filters modified with nanomaterials.


The activity of brain-derived neurotrophic factor (BDF) in the central nervous system has been well-studied, but its physiological role in other organs has not been clearly defined. This review summarizes the current findings on the functionality of BDNF in various peripheral tissues and discusses several unresolved questions in the field.Brain-derived neurotrophic factor (BDNF) is an important growth factor in the central nervous system. In addition to its well-known activities in promoting neuronal survival, neuron differentiation, and synaptic plasticity, neuronal BDNF also regulates energy homeostasis by modulating the hypothalamus’s hormonal signals. In the past decades, several peripheral tissues, including liver, skeletal muscle, and white adipose tissue, were demonstrated as the active sources of BDNF synthesis in response to different metabolic challenges. Nevertheless, the functions of BDNF in these tissues remain obscure. With the use of tissue-specific Bdnf knockout animals and the availability of non-peptidyl BDNF mimetic, increasing evidence has reported that peripheral tissues-derived BDNF might play a significant role in maintaining systemic metabolism, possibly through the regulation of mitochondrial dynamics in the various tissues. This article reviews the autocrine/paracrine/endocrine functions of BDNF in non-neuronal tissues and discusses the unresolved questions about BDNF’s function. In . In 1].  disease . The neu disease ,7,8.Bdnf and Ntrk2 (the TrkB gene) knockout mice revealed that mBDNF participates in the regulation of body weight through controlling food intake i) release [2+]i is crucial in their cellular function, such as cardiomyocytes and the pancreatic β-cells, deleting TrkB.T1 abolished the action of BDNF, which impedes heart contraction and insulin secretion [TrkB.T1 gene results in a greater Ca2+ flux between the cytoplasm and sarcoplasmic reticulum to increase contractility [TrkB.FL and TrkB.T1 as well as their functional interaction in various tissues might represent an additional regulatory mechanism for BDNF response in peripheral tissues. Hence, it would be necessary to consider the role of TrkB.T1 in studying the BDNF’s function in the future.It is also important to determine the dominant receptor of BDNF to exert its metabolic functions in different tissues. The dogma that BDNF only relies on TrkB.FL to initiate the biological activities has been changed since the discovery that TrkB.T1 could transduce intracellular signals by provoking intracellular calcium ([Ca release . This fi release ,136. Howecretion ,137. Becactility . The difLast but not least, peripheral tissues-produced mBDNF might act on the CNS to modulate cognitive functions. When considering together that mBDNF is a well-recognized myokine whose expression is increased after exercise, muscle-derived mBDNF is secreted into the circulation . Althoug


Hepatocellular carcinoma (HCC) is one of the most common cancers worldwide ,2. The eThe practice-changing phase III IMbrave150 trial compared the combination of the PD-L1 inhibitor atezolizumab plus bevacizumab versus sorafenib monotherapy in treatment-naïve patients with advanced HCC ; of noteThe role of another immune-based combination including two ICIs, the PD-L1 inhibitor durvalumab and the anti-CTLA-4 antibody tremelimumab, has been explored in the HIMALAYA trial  21,22,2,222,23. From this point of view, this Special Issue welcomes papers exploring the current state of the art and future perspectives in the immunotherapy of HCC. The Special Issue will aim to assess key open questions in HCC immunotherapy, including preclinical studies, novel immunotherapies and immune-based combinations, biomarkers of response, experimental therapies, real-world experience with immune checkpoint inhibitors, and several other topics.


Sarcoidosis is a multisystem inflammatory granulomatous disease of unknown cause that most commonly affects lungs and lymph nodes, with frequent yet asymptomatic cardiac involvement. The epidemiologically associated cardiovascular risk suggests an underlying prothrombotic state and endothelial dysfunction, currently understudied in the available literature. Therefore, we aimed to investigate prothrombotic plasma properties together with selected echocardiographic and laboratory biomarkers of cardiovascular injury in that disease. N = 53 patients with pulmonary sarcoidosis in clinical remission and N = 66 matched controls were assessed for inflammatory and endothelial injury biomarkers, plasma thrombin generation profile, and echocardiographic and lung function parameters. Sarcoidosis cases had impaired systolic and diastolic left ventricular function, higher concentrations of inflammatory markers, D-dimer and factor VIII activity compared to the controls. The coexistence of extrapulmonary disease was associated with elevated circulating vascular cell adhesion molecule 1, while cases with hypercalcemia had higher thrombomodulin concentration. Sarcoidosis was characterized by the unfavorably altered thrombin generation profile, reflected by the 16% higher endogenous thrombin potential (ETP), 24% increased peak thrombin concentration, and 12% shorter time to thrombin peak in comparison to the control group. ETP was higher in cases with proxies of pulmonary restriction, extrapulmonary–extracutaneous manifestation, and need for corticosteroids use. Despite the clinical remission, sarcoidosis is related to prothrombotic plasma properties and signs of endothelial injury, likely contributing to the higher risk of cardiovascular events. In addition, subclinical cardiac involvement may play an additional role, although further clinical and experimental studies are needed to verify these findings. The disorder most commonly affects the lungs and lymph nodes1. Albeit its etiology is unexplained despite decades of research efforts, studies have shown the role of genetic predisposition, environmental factors, and exposure to unknown infectious agents causing dysregulation of the immune response2.Sarcoidosis is a multisystem inflammatory disease characterized by the formation of microscopic, non-necrotizing structured masses named noncaseating granulomas. They are composed of macrophages, epithelioid cells, giant cells, and lymphocytes, predominantly CD43. Like the pulmonary disease, damage to the heart is caused by acute inflammation and/or tissue fibrosis4, potentially leading to progressive heart failure or death. A recent study indicates that the granulomas formation in the myocardium is related to the increased local expression of oncostatin M and the Reg3A and Reg3γ chemokines involved in the pathogenesis of chronic heart damage5. Since reversible inflammation results in irreversible tissue damage and remodeling, early diagnosis of cardiac involvement is essential for the patient outcome.Although sarcoidosis occurs most often in the nodal-pulmonary form, it can affect other organs, such as skin, eyes, heart, blood vessels, muscular, skeletal, and nervous systems, leading to their dysfunction. The extrapulmonary disease may be initially occult but finally, result in severe complications later in the disease course. In particular, cardiac sarcoidosis is diagnosed clinically in 2–5% of the patients, while autopsy reports indicate the frequency of cardiovascular involvement of about 20–30%; thus, the proper diagnosis is established postmortem6. However, in sarcoidosis patients, the odds of VTE can be over three times higher 8. Furthermore, the involvement of the heart and blood vessels may increase thromboembolic events and sudden cardiac death in affected subjects9. It is estimated that cardiovascular complications account for 13–25% of fatal sarcoidosis cases in the United States. In Japan, where the cardiac form of the disease is more prevalent, this percentage is as high as 58%3.Venous thromboembolism (VTE), including deep vein thrombosis (DVT) and pulmonary embolism (PE), occurs at an incidence of about 1 per 1000 annually in the general adult population10. Inflammatory diseases are associated with an increased risk of thromboembolic complications11, as they contribute to Virchow’s triad: prothrombotic state, endothelial injury, and abnormalities of blood flow12. Inflammatory cytokines, such as tumor necrosis factor alpha (TNF-α) and interleukin (IL)-6, among others, may induce the expression of tissue factor (TF) on the monocytes10. Moreover, reduction of endothelial shear stress at inflammatory loci (as seen in decreased or turbulent blood flow) relates to endothelial proliferation, hypoxia, and release of P-selectin and von Willebrand factor (vWF) from the endothelium, promoting thrombus formation13. In turn, endothelial damage results in nitric oxide, thrombomodulin, and prostacyclin secretion, which to some extent may play a protective role on endothelium, in contrast to endothelin, constricting blood vessels14. Finally, research on sarcoidosis patients demonstrated reduced flow-mediated dilation of the brachial artery, associated with elevated circulating inflammatory biomarkers, such as TNF-α and intercellular adhesion molecule (ICAM)-115. That observation points to endothelial damage, referring to systemic inflammation.The pathogenetic relationship between inflammation and coagulation is well established16. Several studies have previously analyzed the relationship between prothrombotic state, endothelial injury, and chronic inflammation in other lung diseases, including asthma17 and chronic obstructive pulmonary disease (COPD)18; however, no such data is available for sarcoidosis. Since blood coagulation is the common denominator of cardiovascular conditions and thromboembolism which can be modified using contemporary treatment options, its study in non-active sarcoidosis presents an essential scientific task with potential clinical applications.Consequently, acute inflammatory diseases (infections and rheumatologic disorders) are an established risk factor of VTE included in the widely-used Padua prediction score. Furthermore, two observational studies suggested sarcoidosis as an independent risk factor of PE, but did not differentiate between acute and chronic disease19. In addition, Hemker et al. have shown that enhanced thrombin formation in that assay indicates a more extensive and faster activation of the blood coagulation cascade in vitro21. Moreover, further clinical studies have demonstrated that increased plasma thrombin generation is also an indicator of a higher risk of future VTE22, recurrent cardiovascular events23, or stroke in elderly patients24.Calibrated automated thrombin generation (CAT) assay is a well-established, universal, and reproducible tool to measure the entire kinetics of plasma thrombin formation, including its initiation, propagation, and termination25. We have also analyzed its relationship with the inflammatory and endothelial injury biomarkers and lung function and echocardiographic parameters. According to our knowledge, such studies have not been performed to date.Here, we aimed to investigate the plasma thrombin generation profile in non-active sarcoidosis as a potential indicator of the prothrombotic state in these patientsWe have conducted an observational, case–control comparative study of N = 53 white Caucasian adult patients with pulmonary sarcoidosis in clinical remission  who reported to our outpatient clinic in Cracow, Poland, from April 2019 to November 2019, and n = 66 matched controls.26. In each case, the diagnosis was confirmed by histological examination of hilar or mediastinal lymph node biopsy samples, demonstrating non-necrotizing granulomas in the absence of other etiologies . The samples were collected during endobronchial ultrasonography-guided transbronchial needle aspiration or mediastinoscopy. We have staged the disese by the traditional radiological criteria of intrathoracic changes27. Clinical remission was defined as stable lung function, chest radiograph findings, and lack of new respiratory or other disease-related symptoms over the past six months; maintenance therapy with low-dose glucocorticoid  or other immunosuppressants were permitted if the doses were not escalated over the past six months28.The diagnosis of sarcoidosis and the extent of organ involvement was established by a physician according to the criteria of the American Thoracic Society/European Respiratory Society/World Association for Sarcoidosis and Other Granulomatous Disorders (ATS/ERS/WASOG)Control subjects were enrolled from the hospital personnel and their acquaintances or relatives. They were frequency-matched with patients according to age, sex, body mass index (BMI), smoking status and comorbidities  to ensure no statistically significant differences between cases and controls were present.2 using Modification of Diet in Renal Disease [MDRD] study formula) or liver injury .The exclusion criteria for both the study and the control groups included: diagnosis of other autoimmune or pulmonary disease, any acute illness , immobilization or surgical intervention within the past four weeks, pregnancy or breastfeeding, use of oral contraceptives or hormone replacement therapy, known thrombophilia, history of any thromboembolic event, such as myocardial infarction or stroke, coronary artery disease, congestive heart failure, left ventricular (LV) ejection fraction lower than 50%, atrial fibrillation, cancer, current anticoagulation treatment, chronic kidney disease , diabetes mellitus (defined as fasting serum glucose level > 7.0 mmol/l or therapy with anti-diabetic drugs), or hypercholesterolemia  were eligible.The study design was approved by the Bioethics Committee of Jagiellonian University Medical College (protocol number 1072.6120.116.2019). The study was conducted according to the criteria set by the Declaration of Helsinki, and each subject signed informed consent before participating in the study.29.Spirometry, whole-body plethysmography, and diffusing capacity for carbon monoxide were performed in sarcoidosis patients using a MasterLab “Jaeger” equipment following the ATS/ESR Statement30. We used a GE Vivid 7 Dimension Ultrasound and a 4S 2–4 MHz probe with typical parasternal and apical views. Peak mitral inflow E and A velocity waves on pulsed-wave Doppler, E/A ratio, E-wave deceleration time, and isovolumic relaxation time were measured from the apical four-chamber view. In addition, the diastolic e’ velocity was obtained by tissue-Doppler imaging (TDI) at both the septal and lateral mitral origins on a four-chamber apical view. Furthermore, left ventricular (LV) filling indexes (E/e’ ratios) were calculated. LAVI  was calculated by dividing left atrial volume by body surface area.Transthoracic echocardiogram (TTE) was obtained from all patients and control individuals by standard protocolsFasting blood samples were drawn between 7:00 and 11:00 a.m. using minimal stasis. Complete blood cell and platelet count, urea, creatinine, lipid profile, glucose, alanine aminotransferase, and fibrinogen were analyzed by routine laboratory techniques. The estimated glomerular filtration rate (eGFR) was calculated by the Chronic Kidney Disease Epidemiology Collaboration formula (CKD-EPI). C-reactive protein (CRP) was measured by the Cobas Integra System, while factor VIII activity (FVIII:C) by the one-stage clotting assay  and concentrations of D-dimer using the turbidimetric method .Standardized high‐sensitivity immunoenzymatic assays (ELISAs) were used to measure serum levels of IL-6, VCAM-1, and soluble thrombomodulin . In plasma, thrombin-antithrombin complexes (TAT) were investigated by a commercially available immunoenzymatic test .g for 15 min, at room temperature, within 1 h of collection. The platelet-poor plasma was frozen in aliquots and stored at − 70 °C until the thrombin generation assay was analyzed.Moreover, blood samples were drawn into coagulation tubes filled with 0.109 mol/l (3.2%) buffered tri-sodium citrate solution (vol/vol 9:1), and centrifuged two times at 2500×21. A reagent mixture of relipidated recombinant tissue factor (rTF) and phospholipids  was added to the platelet-poor plasma sample (80 μl), followed by the automatic addition of a fresh starting reagent containing calcium chloride (100 mmol/l) and a thrombin-specific fluorogenic substrate (Z-Gly-Gly-Arg-AMC) (2.5 mmol/l) in HEPES buffer. The reaction was completed in a microtiter well . The fluorescence intensity was measured using the Fluoroscan Ascent® fluorometer  using the appropriate software . All the experiments were run in duplicates.Thrombin generation assay was applied using the calibrated automated thrombogram , according to the manufacturer’s instructionspeak thrombin generation —the maximum concentration of thrombin formed during the time of registration,endogenous thrombin potential —the area under the curve showing thrombin formation,lag time (min)—the time from the start of analysis until thrombin starts to generate,time to thrombin peak —the time from the start of thrombin generation until the maximum thrombin concentration is achieved.Four parameters of the thrombin generation curve were analyzed, as shown in Fig. p < 0.05.Statistical analysis was performed using the Statistica version 13.3 package for Microsoft Windows . Categorical variables were provided as percentages, and differences between study groups were analyzed using the χ2 test. The Shapiro–Wilk test assessed the normality of the continuous data distribution. As appropriate, they were presented as medians with a range between 25 and 75th percentiles or means with 95% confidence intervals (CIs) and compared using the Mann–Whitney U test, Kruskal–Wallis, or unpaired t-tests. A one-way analysis of covariance (ANCOVA) was applied to adjust for demographic confounders, i.e., sex, age, BMI, smoking habit, and comorbidities, i.e. arterial hypertension, diabetes mellitus, and hypercholesterolemia. A Spearman rank correlation test was performed to test the associations between continuous variables. Receiver Operating Characteristic (ROC) curves were analyzed to verify optimal cut-off values of ETP, peak TG, lag time, and D-dimer for calculation of odds ratios (ORs) with 95% CIs for differentiation between sarcoidosis and control cases in a multiple logistic regression model. Multiple linear regression models were utilized to indicate factors independently associated with ETP and peak TG. The results were considered statistically significant at a level of Sarcoidosis and control individuals were well matched according to the demographic factors, including age, sex, BMI, smoking habit, and prevalence of comorbidities Table . In TablAlmost half of the patients were treated with systemic corticosteroids with a median duration of 3 years (interquartile range 3–8 years), while only n = 9 (17%) received oral corticosteroids at the time of evaluation Table . Two subAdditionally, due to arterial hypertension, sarcoidosis patients were frequently treated with angiotensin-converting enzyme inhibitors or angiotensin receptor antagonists , beta-blockers , and diuretics . Non-steroidal anti-inflammatory drugs or aspirin were applied by three (5.7%) of them.Results of lung function tests in sarcoidosis patients are also provided in Table Transthoracic echocardiographic measurements revealed that sarcoidosis patients had larger dimensions of the right ventricle, thicker interventricular septum, and posterior LV walls Table . They alp = 0.09, both), patients with the coexisting extrapulmonary manifestation, e.g., extra-thoracic lymphadenopathy, had elevated circulating VCAM-1 as compared to the remaining subjects . Furthermore, those with documented prolonged hypercalcemia were characterized by increased thrombomodulin in peripheral blood .Despite clinical remission, sarcoidosis was characterized by significantly higher concentrations of inflammatory markers, such as CRP, IL-6, and fibrinogen. These patients also had slightly elevated D-dimer, FVIII:C, red blood cell and platelet count, lower lymphocyte count, increased triglycerides, and high-density lipoprotein cholesterol levels Table . Moreovep = 0.003), and both of them were related to the plasma D-dimer .Concentrations of thrombomodulin and VCAM-1 correlated well with each other , which also remained in a positive relationship with plasma thrombomodulin . Furthermore, CRP and IL-6 were related to the D-dimer .As expected, CRP was associated with IL-6  and 12% faster ttPeak  as compared to controls .Consequently, a predictive logistic regression model based on D-dimer, ETP, and ttPeak was able to discriminate between sarcoidosis patients and the control group with ROC AUC of 0.851 (AUC error of 0.0431) and cut-off points of 275 ng/ml, 1737 nmol/l × min, and 5.38 min, for D-dimer, ETP, and ttPeak, respectively .Thrombin generation parameters were independent of demographic factors, including sex, age, and BMI in both studied groups. In turn, among basic laboratory measures, only triglycerides were linked with CAT assay variables, such as ETP and peak TG . The number of affected organs was not correlated with thrombin generation parameters.Subgroup analysis revealed that neither radiological stage of the disease, symptoms at diagnosis, past inhaled corticosteroid, nor immunosuppressant use was related to the altered thrombin generation profile. In turn, ETP was higher in those with extrapulmonary–extracutaneous manifestation, including optic nerve, uvea, nasal septum and/or liver involvement , while the patients using corticosteroids at assessment had elevated peak TG as compared to those currently untreated .Furthermore, patients on systemic corticosteroids, currently or in the past, were characterized by enhanced thrombin formation in CAT assay. The patients treated in the past had higher ETP as compared to those never treated  and vital capacity (VC) before bronchodilator . Consequently, ETP was significantly higher in the only four patients  with TLC below the normal range, as compared to the remaining sarcoidosis individuals .Interestingly, in sarcoidosis, ETP was inversely correlated with total lung capacity .Regarding echocardiographic measures, we found a positive association between ETP and systolic pulmonary artery pressure  on inflammatory, epithelial, and endothelial cells in the lungs. PARs are triggered by several proteases, including neutrophil elastase, granzymes of cytotoxic lymphocytes, and thrombin, enhancing local inflammatory response44. Moreover, it has been documented that activating PAR-1 on monocytes and macrophages upregulates the production of oncostatin M, a pleiotropic cytokine implicated in the pathology of the heart and vascular damage45.Our results do not explain how lung inflammatory granulomas drive the prothrombotic state. One possible explanation might be related to the activity of Rac-1, a small G-coupled protein, a molecular switcher regulating cell adhesion, proliferation, migration, and chemotaxis. It has been demonstrated that Rac-1 implies T-cell and macrophage to sarcoid granulomas formation46. Therefore, it may underline the importance of strict dyslipidemia management in sarcoidosis patients. In turn, the inverse relationship between thrombin generation and kidney function is well-known and mirrors other reports48.As expected, increased ETP was related to the higher serum triglycerides. However, in our study, that association was more potent than previously reported in the general population49. Therefore, one may speculate that increased VCAM-1 in our study does not necessarily indicate vascular damage. Instead, its higher level, interestingly related to the lower lymphocyte count in peripheral blood (data not shown), may reflect vascular susceptibility to the lymphocyte homing into the sarcoid lymph nodes.Another important issue that merits comment is a higher circulating VCAM-1 in sarcoidosis patients with extrapulmonary lymphadenopathy. VCAM-1 is an endothelial cell biomarker, which appears on these cells after activation. Therefore, its higher blood concentration might suggest endothelial injury. However, it has been previously shown that increased VCAM-1 on endothelial cells is the first step of signaling mechanisms in lymph nodes, leading to lymphocyte accumulation and lymph nodes enlargement2D3 by activated macrophages50. Of note, the active form of vitamin D3 stimulates osteoblasts to synthesize thrombomodulin51. Thus, higher thrombomodulin in hypercalcemia might also be related to the macrophage stimulation in sarcoid granulomas.In turn, the second analyzed endothelial damage biomarker, thrombomodulin, was higher in patients with hypercalcemia. Hypercalcemia affects about 20% of sarcoidosis individuals. The majority of those cases can be explained by the overproduction of 1,25(OH)53. It is related to heart wall relaxation disorders, LV mass hypertrophy, and myocardial fibrosis, leading to increased LV filling pressure, left atrium pressure, and, consequently, pulmonary hypertension, which ultimately causes clinical symptoms54. Additionally, a postulated mechanism underlying cardiac diastolic failure is endothelial dysfunction of the coronary microcirculation due to increased circulating inflammatory cytokines 56. Furthermore, coronary circulation's endothelial damage likely induces cardiomyocyte hypertrophy and increased collagen production in interstitial tissue57.The last issue for discussion in our data is echocardiographic findings. In comparison to the controls, the patient group had enlarged dimensions of the right ventricle, left atrium, and thicker LV walls. Furthermore, sarcoidosis was related to diastolic cardiac dysfunction and the increased probability of pulmonary hypertension. Apart from cardiological cases, diastolic LV impairment is often demonstrated in pulmonary patients, particularly those with COPD or sleep apnea syndrome58. While cardiac sarcoidosis is asymptomatic in 95% of the affected patients, untreated inflammation and progressive myocardial fibrosis can lead to heart chamber abnormalities and dysfunction. It is possible that the described diastolic dysfunction may be a consequence of undetected cardiac sarcoidosis. Of note, the diagnosis of this abnormality is difficult. Identified electrocardiographic or echocardiographic findings can often be explained by comorbidities such as hypertension or coronary heart disease3. At the same time, the sensitivity and specificity of the standard two-dimensional (2D) echocardiography are limited. However, new techniques, such as speckle tracking echocardiography, cardiac magnetic resonance, and positron emission tomography, can be beneficial in the early detection of inflammatory and fibrotic changes of the heart and thus need to be recommended in patients with suspected cardiac sarcoidosis61.Documented echocardiographic features seem clinically insignificant; however, they might increase the risk of cardiovascular events in sarcoidosis. Moreover, the elevated left atrial volume index has been shown as an independent predictor of adverse cardiovascular outcomes62. Therefore, sarcoidosis patients on systemic corticosteroids might be at increased risk of VTE63. Finally, we did not determine other potential modulators of coagulation, e.g., genetic variants. Given prothrombotic abnormalities in circulating blood of first-degree relatives of patients with venous thrombosis64, we cannot exclude that a documented prothrombotic state is genetically determined, at least to some extent.Our study has several limitations. First, the study sample was relatively small; in particular, subgroup analysis should be interpreted with caution. Second, laboratory measures were analyzed once, and we cannot exclude their changes over time. Moreover, our observational study has no follow-up; thus, we cannot verify if enhanced plasma thrombin generation refers to the increased risk of VTE episodes. Third, sarcoidosis patients had internal comorbidities that may affect thrombin generation assays. However, we have matched controls for them and adjusted statistics in the ANCOVA analysis. Thus, we believe that the primary outcomes presented here are reliable. Also, the effect of the medication used was beyond the scope of our study. However, higher peak TG in patients currently treated with systemic corticosteroids is an interesting finding, possibly related to those medications' unfavorable side effects, as previously shown in healthy individualsThe current study is the first to show increased in vitro plasma thrombin generation in patients with sarcoidosis, which might indicate the prothrombotic state in that disease. However, further extensive observational and prospective studies are needed to verify and explore that topic.


Graphene and its derivatives have been a burning issue in the last 10 years. Although many reviews described its application in electrochemical detection, few were focused on food detection. Herein, we reviewed the recent progress in applying graphene and composite materials in food detection during the past 10 years. We pay attention to food coloring materials, pesticides, antibiotics, heavy metal ion residues, and other common hazards. The advantages of graphene composites in electrochemical detection are described in detail. The differences between electrochemical detection involving graphene and traditional inherent food detection are analyzed and compared in depth. The results proved that electrochemical food detection based on graphene composites is more beneficial. The current defects and deficiencies in graphene composite modified electrode development are discussed, and the application prospects and direction of graphene in future food detection are forecasted. It has Graphene oxide (GO), an oxygen-containing derivative of graphene, belongs to a branch of graphene research. Generally, graphene oxide is a material obtained by the multistep chemical or thermal reduction of graphene  Dikin e. ChemicaReduced graphene oxide (rGO) is an ideal support material for metal nanoparticles, which can be produced at a low cost and large scale. rGO sheets possess higher conductivity than GO sheets due to restoring the conjugated network in the rGO sheets. The oxygen-containing functional groups on rGO render it with an extremely high specific surface area, superior electronic conductivity, excellent mechanical strength, and elasticity . rGO is Graphene quantum dots (GQDs) are attractive nanomaterials consisting of a monolayer, or a few layers, of graphene with excellent and unique properties. Unlike graphene sheets, GQDs are 0D graphene segments that exhibit bandgap responsible for their unique optical and electrical properties. Due to their small size, GQDs display a quantum effect when produced using carbon-rich precursors such as fullerene, glucose, graphite, graphene oxide (GO), CNTs, and carbon fibers (CFs). Two main GQD synthesis methods are followed, namely, top–down and bottom–up methods, as shown in 2 substrate. Their research showed that the resulting graphene structure was more homogenous than when using other methods.Graphene sheets are removed directly from the surface of large graphite crystals by mechanical force and transferred to carriers such as silica. Monolayer graphene sheets can be obtained on substrates such as silica. This is the first known method to synthesize graphene using a top–down technique, by which layered graphene is split into several layered graphene sheets through repeated peeling. The obtained flakes varied greatly in size and thickness, and the product size could not be controlled. Therefore, it could not meet the industrial requirements of graphene. Entani et al. developed and modified these methods by synthesizing graphene on a SiO6 Pa), silica atoms are volatilized, and the residual carbon atoms are arranged on the SiC surface to form graphene layers throughout the structure.Under high vapor pressure, high temperature , and ultra-high vacuum pressure  are used to treat the raw graphite. Small molecules of a strong acid are inserted in between the graphite layers, followed by strong oxidants such as KMnO4 and KCIO4  . Currentnd KCIO4 . Finallybtain GO . The degbtain GO .via CVD is of high quality and is expected to meet the application requirements of transparent conductive films. It is widely used in the large-scale industrial preparation of semiconductor materials for thin-film transistors.The graphene prepared As a branch of chemistry, electrochemical analysis has been verified by many practical and theoretical experiments. It is an analytical method to determine the composition and content of an object by directly measuring different electrical signal parameters, such as the current potential conductance, according to the electrochemical characteristics of different substances. Electrochemical detection methods have the advantages of low cost, small working space, fast speed, high sensitivity, and high accuracy when compared with other food detection methods. Therefore, the research, application, and promotion of electrochemical detection in food quality and related areas can play an essential role in promoting future food quality detection and standards. As an eminent factor in materials science in the 21st century, graphene has been widely used according to its excellent properties . ApplyinThe synthetic pigments that commonly appear in food are azo pigments . Azo dyes account for more than half of the global synthetic dye production. Azo pigments exhibit excellent color performance, and desired colors can be obtained by adjusting the type and proportion of the azo components, creating a wide variety of colors. The excessive intake of azo dyes may be harmful to human health. For example, sunset yellow and tartrazine contain groups such as azo and aromatic rings in their structure, which can induce allergies, asthma, migraines, and cancers . Existin2, to develop a rapid and low-cost method without any sample pre-treatment required to detect sunset yellow and tartrazine yellow colorants, with the detection limits of 6.0 and 8.0 nM(2 nanorods (MnO2 NRs) with a modified glassy carbon electrode (GCE) surface. When compared with standard GCE, a well-defined sunset yellow (SY) oxidation peak was observed at the MnO2 NR-ERGO/GCE. It had a detection limit of 2.0 nM and a good linear response to the SY in the ranges of 0.01–2.0 µM, 2.0–10.0 µM, and 10.0–100.0 µM. This method was applied to detect SY in soft drinks and obtained satisfactory results . Palanisamy et al. fabricated graphene/β-CD/PtNP-modified electrodes through platinum nanoparticles (PtNPs) decorated with graphene-β-cyclodextrin (graphene/β-CD)–modified electrodes. Studies on the electrode cyclic voltammetry determination of different modified electrodes on the electrochemical behavior of Sudan I indicated that the electric catalytic activity of graphene/β-CD/PtNP-modified electrodes on Sudan I was higher than that of other modified electrodes. With a detection linear response range of 0.005–68.68 µM and a detection limit of 1.6 nM, the sensor was used for monitoring Sudan I–adulterated food samples  and achieved satisfactory results in practical tests 6] 3+/Nafion via the drop coating method on a glassy carbon electrode (GCE), which successfully determined trace amounts of Pb(II), Cu(I), Cu(II), Sn(IV), and Hg(II) in canned food samples. Pb2+, Cd2+, and Cu2+ exhibited the highest sensitivity by anodic stripping voltammetry (ASV). There was an excellent linear relationship between heavy metal concentration and peak current, and the detection limit of Pb2+ was 0.74 ppb, that of Cd2+ was 37 ppb, and that of Cu2+ was 945 ppb  added to food samples. Karthik used electron microscopy (HR-SEM), transmission electron microscopy (TEM), X-ray diffraction (XRD), and UV–Vis-NIR Fourier-transform infrared spectroscopy (FT-IR) to analyze cobalt-doped zinc oxide/reduced GO (Co: ZnO/RGO) nanorods, obtained by chemical co-precipitation  100–2,500 μM, with a sensitivity of 0.8587 μAμM−1 cm2. The detection limit was 0.5 nM (S/N = 3). The method successfully determined methyl parathion in ugli and tomato fruits, beetroot, and broccoli, indicating its practical applicability (Organophosphate pesticides (OPs) have been widely used in agricultural production due to their wide control range, low cost, and high insecticidal efficiency. Organophosphates are neurotoxins that inhibit acetylcholinesterase activity, causing acetylcholine accumulation and neurotoxicity . In the cability .In the early 1950s, methylcarbamate was introduced as a pesticide and is still used today . This is2) nanocomposite. As-synthesized GOS@CuFeS2 nanocomposite film screen-printed carbon-modified electrode (SPCE) displays excellent electrocatalytic ability toward MOX. Under optimized working conditions, the modified electrode provides a linear response range from 0.073 to 801.5 µM. The detection limit was obtained as 4.5 nM. The sensor displayed outstanding sensitivity at 17.97 μA μM−1 cm−2 35. The GOS@CuFeS2 nanocomposite–modified electrode shows greater real-time practicality in actual vegetable samples (Methyl paraoxon (MOX) is a highly toxic organophosphate pesticide. It has been recently reported that MOX can enter the human body through ingestion, inhalation, or dermal penetration. Due to its high nondegradability, it can bind to fruits and vegetables. When consumed, it can impose sub-chronic and chronic diseases by inhibiting acetylcholinesterase in human metabolism. Umamaheswari et al. reported the detection of non-enzymatic electrochemical sensors based on the 3D porous phase graphene oxide sheet–encapsulated chalcopyrite –coated glass substrates through electrophoretic deposition. The AFB1 monoclonal antibody was covalently fixed on the deposition electrode GQDs/ITO , improved detection limit (0.23 ng ml−1), and good stability (5 weeks) in label-free detection (via electrophoretic deposition. The sensing results of the anti-AFB1/RGO/ITO––based immunoelectrode obtained as a function of aflatoxin concentration showed high sensitivity (68 mA ng−1 ml cm2) and an improved detection limit (0.12 ng ml−1) (One of the most harmful food contaminants is aflatoxin (AFT) . AflatoxGQDs/ITO . The det in food . Srivastetection . Pumera etection . Reducedetection . Srivastng ml−1) .n = 6), and stability. Subsequently, it has been successfully applied in maltol analysis in various foods . Maltol is banned in children’s food by both the FDA and China’s national food safety standards because of its potential to harm health and is also banned in Europe . Therefous foods . Gan’s sus foods .3O4/polyDOPA/RGO composite material using a green method, and its electrocatalytic activity on nitrite oxidation was excellent  and high relative recovery (82.70–105.97%) (Acrylamide (2-propenamide) is found in carbohydrate-rich food cooked under high temperatures. Nodeh et al. (2018) successfully cleaned and measured acrylamide in various foods using a Fe105.97%)  (Nodeh e105.97%) .2Te3@g-C3N4 BNs to quantify food toxic chemicals in meat samples. As modified, the Bi2Te3/g-C3N4 BN-modified electrode exhibits excellent electrochemical activity toward food toxic ractopamine (RAC) with high-sensitive (L.R: 0.015–456.4) and nanomolar detection limit (LOD: 1.77 nM)  synthesized a super-active electrocatalyst of Bi1.77 nM) .2 NSs). Features including (a) a wider linear range (0.05–490 μM), (b) a nanomolar detection limit (0.03 μM), and (c) high sensitivity (29 μA μM−1 cm−2) have been recorded. It also yields high accuracy and good recovery  developed a sensitive electrochemical  sensor for the determination of coccidiostat drug (roxarsone) based on the use of an SPCE (screen-printed carbon electrode) modified with tungsten disulfide nanosheets . The electrode has a high level of selectivity in the large excess concentrations of interfering species. In addition, the modified electrode offers satisfactory repeatability, reproducibility, and stability (4 embedded on sulfur-doped–graphitic carbon nitride composites (SrMoO4/SGCN) using green methods for the electrochemical detection of CAP. The sensor can be used in river water samples, urine, and human blood serum with a high recovery rate. SrMoO4/SGCN/GCE impedance shows a lower resistance charge transfer (Rct), which helps favor the superior electrochemical detection of CAP. The SrMoO4/SGCN/GCE exhibits an ultralow detection limit of 1.5 nM for an extensive concentration range of 0.005 1316.8 M and high sensitivity is 9.619 AM/cm2 using the amperometric method  over a range of 0.02 374.4 μM, with a nanomolar detection limit of 1.2 nM   array electrochemical sensor was used to detect sulfadiazine (SM2) residues in food. Under optimized conditions, a wide linear range of 0.2–1000 ng/ml and a detection limit of 0.169 ng/ml (S/N = 3) were obtained. The recovery rate of the sensor is 92.3–102.23%, and the relative standard deviation is 2.27–4.10%  . Wei’s s27–4.10% . He and 27–4.10% .2Te3) decorated graphitic carbon nitride (GCN) nanostructures as a modified electrode for electrochemical sensing. A nanomolar limit of detection (1.36 nM) was calculated in a 0.05-M phosphate buffer (PB) supporting electrolyte (pH 7.0) using differential pulse voltammetry. The linear dynamic ranges concerning salbutamol concentration were 0.01 892.5 μM, and the sensor’s sensitivity was 36.277 μA μM−1cm2.Rajaji et al. developed a rapid detection of the feed additive drug  using bismuth telluride (BiHz/60 W). As-prepared ZnS NPs@RGO hybrid was applied toward the electrochemical determination of caffeic acid (CA) in various food samples. The sensor can be used in red wine and soft drink samples with a high recovery rate. The proposed electrochemical caffeic acid sensor produces a wide linear range of 0.015–671.7 µM with a nanomolar level detection limit (3.29 nM)  encapsulated on reduced graphene oxide (RGO) hybrid by an ultrasonic bath (50 k3.29 nM) .Graphene is an excellent new 2D material with an extensive application range . HoweverAt present, the biodegradation of graphene materials mainly focuses on enzyme degradation. During body metabolic processes, the biological effect of hydrogen peroxide during substance oxidative decomposition is combined with active enzymes in the body to achieve the biodegradation of graphene materials. Differences in graphene (and its derivatives) structure, properties, and composition affect their degradation behavior. Using polyphase atom doping and surface functionalization, the physical and chemical properties of the materials are changed to affect the degradation process to regulate the enzymatic degradation of graphene materials. Some methods of obtaining graphene cannot meet research specifications due to dangerous pollution levels and low purity. Therefore, the development of green and environmentally friendly preparation methods is needed.Based on existing electrochemical detection models, adding modified functional graphene-like materials to develop new, more reliable, rapid, and accurate detection methods and improving detection stability and limits has always been an essential topic in food detection. Although the commercialization of graphene materials is still in its infancy, the related long-term safety and environmental issues still need to be addressed. The physical and chemical properties of graphene have a decisive influence on biological outcomes, such as the effects of size, shape, surface charge, chemical composition, and surface modification on the toxicity of biofilm, which are still unknown. Furthermore, the active mechanisms are still unclear. Research on the mechanisms and biology of graphene interaction is still in its infancy at home and abroad. The physiological and biochemical processes caused by graphene’s impact on organisms have not been thoroughly studied. Therefore, it is urgent to study further the toxic effects and functional mechanisms of graphene biofilms.


Pmmn borophene. We also show that smooth electron waveguides in tilted Dirac materials can be used to manipulate the degree of valley polarization of quasiparticles travelling along a particular direction of the channel. We examine the particular case of the hyperbolic secant potential to model realistic top-gated structures for valleytronic applications.We show that if the solutions to the (2+1)-dimensional massless Dirac equation for a given one-dimensional (1D) potential are known, then they can be used to obtain the eigenvalues and eigenfunctions for the same potential, orientated at an arbitrary angle, in a 2D Dirac material possessing tilted, anisotropic Dirac cones. This simple set of transformations enables all the exact and quasi-exact solutions associated with 1D quantum wells in graphene to be applied to the confinement problem in tilted Dirac materials such as 8- A broad class of quasi-1D potentials can also be solved quasi-exactly by transforming the 2D Dirac equation into the Heun equation2 or one of its confluent forms5, or via the application of Darboux transformations11. These exact and quasi-exact bound-state solutions have direct applications to electronic waveguides in 2D Dirac materials13, such as graphene, where the low-energy spectrum of the charge carriers can be described by a Dirac Hamiltonian14, and the guiding potential can be generated via a top gate20. Recent advances in device fabrication, utilizing carbon nanotubes as top gates, has enabled the detection of individual guided modes21, opening the door to several new classes of devices such as THz emitters13, transistors12, and ultrafast electronic switching devices22. These advances in electron waveguide fabrication technology make the need for analytic solutions all the more important, since they are highly useful in: determining device geometry, finding the threshold voltage required to observe a zero-energy mode12, calculating the size of the THz pseudogap in bipolar waveguides13, as well as ascertaining the optical selection rules13 in graphene heterostructures.It can be shown using supersymmetric methods that whenever the Schrödinger equation can be solved exactly for a one-dimensional (1D) potential, there exists a corresponding potential for which the two-dimensional (2D) Dirac equation admits exact eigenvalues and eigenfunctions23. There are only a handful of 2D electronic systems that have been predicted to host these tilted cones33, one of which is 8-Pmmn borophene34, which has attracted considerable attention. In general, boron-based nanomaterials are a growing field of interest37; indeed, exploring the tilt of 2D Dirac cones in the context of 8-Pmmn borophene has recently led to a plethora of theoretical works spanning many fields of research84 from optics to transport, and many more; for a comprehensive review, see Ref.85. The spectacular rise of borophene, and the growing interest in tilted Dirac materials, has led to the revisiting of several well-known problems in graphene, e.g., Klein tunneling47 and transport across quasi-1D heterostructures86, within the context of tilted Dirac materials. As mentioned previously, methods such as supersymmetry and reducing the Dirac equation to the Heun equation, utilize solutions of known problems, to generate solutions to new ones. This begs the question: does a simple mapping exist which would allow us to harness the large body of exact and quasi-exact solutions for 1D waveguides in graphene and then apply them to materials with tilted Dirac cones?In extension to the well-known case of graphene, Dirac cones can in general possess valley-dependent tilt14, for the same potential, but of modified strength, effective momentum, and modified energy scale. After outlining the transformation, we study the particular case of the hyperbolic secant potential, which in graphene is known to admit quasi-exact solutions to the eigenvalue problem; but nevertheless, the whole spectrum can be obtained via a semi-analytic approach12. We use this known graphene waveguide spectrum to generate the corresponding tilted waveguide spectrum, which is verified using a transfer matrix method. Finally, we discuss valleytronic applications.In what follows, we show that the differential equations governing guided modes in an anisotropic tilted Dirac waveguide (orientated at an arbitrary angle) can, via a simple transformation, be mapped onto the graphene problem, i.e., transformed into the massless 2D Dirac equationK and Pmmn borophene23, 2B:Pmmn borophane87, and 25. In what follows, we set The Hamiltonian describing the guided modes contained within a smooth electron waveguide in a tilted Dirac material can be written asHence, the wave vector operators in the non-rotated coordinate system A and B sublattices of the tilted Dirac material. These coupled first-order differential equations can be recast into the same equations used to describe guided modes propagating along a smooth electron waveguide in graphene:L is a constant, associated with the effective width of the potential. We define the tilt and anisotropy parameters as The Hamiltonian, Eq. , acts onentclass1pt{minimat with T with It should be noted that to perform the same transformations for the other chirality  used in the mapping. In the same figure, we also plot the numerical solutions to the tilted Dirac problem obtained via a transfer matrix method . We show in blue crosses the exact solutions given in Eq. , smooth electrostatic potentials in tilted Dirac materials can be utilized as a means to achieve valley polarization. The majority of studies have focused on tunneling across electrostatically-induced potential barriers, and valley filtering and beam splitting have been demonstrated86. It has also been shown that the allowed transmission angles through a potential can be controlled using magnetic barriers86. We propose a change in geometry: rather than studying chirality-dependent transmission across barriers, we shift the focus to studying guided modes along quasi-1D confining potentials. The conductance along such a channel can be measured by placing one terminal at each end. According to the Landauer formula, when the Fermi level is set to energy E (by modulating the back-gate voltage21), the conductance along the waveguide is simply K and It has been suggested that the valley quantum number can be used as a basis for carrying information in graphene-based devices21. Full valley polarization can be achieved for energies less than the lowest-lying supercritical state , for a 2D Dirac material hosting anisotropic, tilted Dirac cones. Therefore, all the rich physics associated with guiding potentials in graphene, e.g., THz pseudogaps in bipolar waveguides, can be revisited in the context of tilted Dirac materials, but with the distinct advantage of knowing the eigenvalues and eigenfunctions. We have also shown that in stark contrast to smooth electron waveguides in graphene, valley degeneracy can be broken in tilted 2D Dirac materials for a broad range of waveguide orientations, anisotropy and tilt parameters. The degree of valley polarization along the waveguide can be controlled by varying the potential strength of the top gate, and also by changing the back-gate voltage. Tilted 2D Dirac materials, such as 8-The Supplementary Information contains a full description of the transfer matrix method used to calculate the band structure of guiding potentials in 2D Dirac materials with tilted Dirac cones.Supplementary Information.


In sarcoidosis, peripheral lymphopenia and anergy have been associated with increased inflammation and maladaptive immune activity, likely promoting development of chronic and progressive disease. However, the molecular mechanisms that lead to reduced lymphocyte proportions, particularly CD4+ T-cells, have not been fully elucidated. We posit that paradoxical peripheral lymphopenia is characterized by a dysregulated transcriptomic network associated with cell function and fate that results from altered transcription factor targeting activity.Messenger RNA-sequencing (mRNA-seq) was performed on peripheral blood mononuclear cells (PBMCs) from ACCESS study subjects with sarcoidosis and matched controls and findings validated on a sarcoidosis case-control cohort and a sarcoidosis case series. Preserved PBMC transcriptomic networks between case-control cohorts were assessed to establish cellular associations with gene modules and define regulatory targeting involved in sarcoidosis immune dysregulation utilizing weighted gene co-expression network analysis and differential transcription factor involvement analysis. Network centrality measures identified master transcriptional regulators of subnetworks related to cell proliferation and death. Predictive models of differential PBMC proportions constructed from ACCESS target gene expression corroborated the relationship between aberrant transcription factor regulatory activity and imputed and clinical PBMC populations in the validation cohorts.We identified two unique and preserved gene modules significantly associated with sarcoidosis immune dysregulation. Strikingly, increased expression of a monocyte-driven, and not a lymphocyte-driven, gene module related to innate immunity and cell death was the best predictor of peripheral CD4+ T-cell proportions. Within the gene network of this monocyte-driven module, TLE3 and CBX8 were determined to be master regulators of the cell death subnetwork. A core gene signature of differentially over-expressed target genes of TLE3 and CBX8 involved in cellular communication and immune response regulation accurately predicted imputed and clinical monocyte expansion and CD4+ T-cell depletion.Altered transcriptional regulation associated with aberrant gene expression of a monocyte-driven transcriptional network likely influences lymphocyte function and survival. Although further investigation is warranted, this indicates that crosstalk between hyperactive monocytes and lymphocytes may instigate peripheral lymphopenia and underlie sarcoidosis immune dysregulation and pathogenesis. Future therapies selectively targeting master regulators, or their targets, may mitigate dysregulated immune processes in sarcoidosis and disease progression. Sarcoidosis is a systemic inflammatory disease characterized by dysregulated immune processes . The intHistorically, lymphopenia has been considered the product of hypersplenism, bone marrow infiltration, or T-cell sequestration , 9–11. HDespite these implications, the underpinnings of peripheral lymphopenia in sarcoidosis remain unclear and comprehensive assessment of the genetic repertoire associated with this phenomenon has been limited . To dateBased on evidence from our prior studies, we hypothesized that a dysregulated transcriptomic network characterizes circulating immune cells in sarcoidosis and aimed to discern master regulator transcription factors of aberrant gene expression to gain insight into the molecular mechanisms underlying paradoxical peripheral lymphopenia in sarcoidosis , 16, 43.in-silico cellular deconvolution. Further details pertaining to study cohorts, mRNA-seq library preparation and annotation for the ACCESS cohort, processing of raw microarray expression data for the UCSF cohort, and mRNA-seq library preparation and annotation for the UIC STAR cohort are described at length in the data supplement. All raw sequencing data for the ACCESS cohort has been deposited to GEO, series record GSE155644. Study approval was obtained through the UIC institutional review board (Protocol #s: 2019-0452 and 2016-0063). Briefly, upon messenger RNA extraction from peripheral blood mononuclear cells (PBMCs), gene expression data obtained from whole transcriptome shotgun sequencing (mRNA-seq) was investigated in treatment naïve Caucasian subjects newly diagnosed with sarcoidosis (n=14) and matched controls (n=14). All 28 subjects were enrolled through the RNA core laboratory in “A Case Controlled Etiologic Study of Sarcoidosis”  as previously described and utilized as a reference cohort in this study given that the gene expression profiles from cases were deemed to represent baseline immune dysregulation in sarcoidosis . InclusiWGCNA) to identify gene modules or clusters and their relationship with sarcoidosis and other clinical features  test or linear regression models and considered significant if p-value < 0.05. Differential expression of genes between sarcoidosis cases and controls within modules significantly associated with disease in the ACCESS cohort was determined utilizing a moderated t-statistic at a predetermined false discovery rate (FDR) of 10%  (https://david.ncifcrf.gov)  was utilized to determine whether specific PBMC subset enrichment was associated with gene expression within modules of interest . Further details regarding network construction, module preservation, and cell subset enrichment are provided in the data supplement.Module eigengenes, obtained from specific gene expression of standardized modules between the ACCESS and UCSF cohorts, were extracted as representations of modular gene expression. Associations between gene modules and clinical features  were evaluated by Spearman’s correlation and ue <0.1) . Subsequcrf.gov) , 51. Biointerest , 53. DifMonster algorithm  within a gene regulatory network. To determine DTFI that underlie immune regulatory networks in health and disease, ACCESS cohort control samples (n=14) were defined as the baseline (reference) state and sarcoidosis samples (n=14) as the perturbed  state. DTFI significance was established if the false discovery rate (FDR) p-value based on the Monster Z-score method was <0.1.To identify master regulator transcription factors capable of driving the changes in PBMC transcriptional programming (cell state transitions) from the healthy to the dysregulated immune response observed in sarcoidosis we applied the lgorithm . MonsterMonster algorithm were extracted separately for the ACCESS cohort sarcoidosis cases and controls. Then bipartite networks were filtered to include significant DTFI and differentially expressed target genes with greater predicted edge weight in sarcoidosis compared to controls. Additional subnetwork reduction was performed to evaluate significantly enriched GO biologic processes associated with cellular proliferation or death assumed to alter PBMC subset proportions. Specifically, only target genes related to GO biologic processes involved in cell “aging/senescence,” “apoptosis,” “arrest,” “autophagy,” “cycle,” “death,” “differentiation,” “division,” “exhaustion,” “phagocytosis,” “proliferation,” and “quiescence” comprised the final subnetworks. Upon subnetwork construction, the Laplacian centrality measure for directed and weighted networks was utilized to perform topological analysis and determine transcription factor relevance  of receiver operating characteristic (ROC) curves determined by the model’s ability to accurately predict “high” or “low” PBMC subset proportions in comparison to negative controls using Venkatraman’s test  scores were calculated and utilized to determine the discriminatory power of each feature within PLS-DA models. Individual target genes with high VIP scores (>1) were considered the most relevant predictors of cell subset variability in sarcoidosis  were constructed using data from the ACCESS cohort for model calibration and externally validated . To corrn’s test . Lastly,coidosis , 60.Our aim was to examine gene expression and transcriptional regulation patterns that might contribute to altered peripheral immunity in sarcoidosis. We obtained PBMCs from subjects with sarcoidosis within 6 months of diagnosis and their matched controls (n=14 per group) in the ACCESS cohort and performed mRNA-seq and network-based analyses. Baseline characteristics of the ACCESS cohort are outlined in 2 test p-value >0.05). Significant differences were found with regards to age between cohorts. Specifically, UCSF cohort cases and controls were found to be significantly older than their ACCESS cohort counterparts  with the majority of subjects in age groups 4 and 5 . UCSF cohort sarcoidosis cases included 14/31 (45%) cases with low lung function defined by the study’s original parameters as forced expiratory volume in one second (FEV1) or forced vital capacity (FVC) percent predicted <80%. Individual subject data pertaining to other variables  was not publicly available. Nonetheless, in contrast to the ACCESS cohort, the original GSE19314 cohort included only 2/38 cases with a positive smoking history, 10/38 with Scadding stage 4 sarcoidosis, and at least 32% on systemic therapy for sarcoidosis.A publicly available PBMC gene expression data set (GSE19314) was used to validate expression and transcriptional regulation analyses performed in the ACCESS cohort Table 2 2 test p-value >0.05) but UIC STAR cohort subjects tended to be older than ACCESS counterparts . Significant differences in smoking history were not observed between the UIC STAR and ACCESS cohorts . Furthermore, pulmonary function between the UIC STAR and ACCESS cohorts was similar with only FEV1% predicted showing a trend towards significantly lower values in the UIC STAR cohort  despite the fact that the majority of subjects in the UIC STAR cohort had chronic disease (>3 years duration), 50% (15/30) had severe disease, and only 20% (6/30) were off of systemic therapy. Other differences between the UIC STAR cohort and UCSF cohort could not be assessed due to lack of publicly available data.Unlike the Caucasian case-control cohorts, the UIC STAR cohort (n=30) consisted of a predominantly African American (66.7%) case series of subjects with sarcoidosis as detailed in the data supplement Table 3.WGCNA, 12 distinct modules of gene expression were identified within the 28 subjects sampled in the ACCESS cohort  and one trended towards significance     but had no association with disease status. Thus, expression of unique genes within the modules significantly associated with disease was considered independent of age, gender, and smoking history.To analyze disrupted patterns of gene expression in sarcoidosis, mRNA-seq data obtained from the ACCESS cohort and microarray data from the UCSF cohort were filtered and normalized to identify 12,047 genes commonly expressed within both datasets. Utilizing BH.adj p-value <0.1; respectively) and likely attributable to clinical, demographic, and geographic heterogeneity between the cohorts  gene ontology terms that characterized the magenta module were consistent with fundamental notions of sarcoidosis and included: “immune system processes (GO:0002376),” “defense response (GO:0006952),” “immune response (GO:0006955),” “regulation of immune system processes (GO:0002682),” and “cell activation (GO:0001775)”. In contrast, the blue module was characterized by significant enrichment of “RNA metabolic processes (GO:0016070),” “nucleic acid metabolic processes (GO:0090304),” “gene expression (GO:0010467),” “nucleobase-containing compound metabolic processes (GO:0006139),” and “cellular aromatic compound metabolic processes (GO:0006725)” suggesting that sarcoidosis is not only marked by immune dysregulation but may also be a result of altered metabolic pathways. Other significantly enriched ontology terms related to multiple innate immunity and cellular immunity pathways and genes were also found in the magenta and blue modules, respectively  , 51. TheTo determine if module-specific gene expression was driven by distinct immune cell types, absolute immune signal (ABIS) deconvolution was performed utilizing the Human Protein Atlas (HPA) and demonstrated that the magenta module was significantly associated with protein coding genes characteristic of monocytes and dendritic cells. The magenta module was therefore renamed the monocyte activity module (MAM) , 53. TakMonster to independently infer master regulators driving cell state transcriptional programs and their target genes in MAM and LSM within the ACCESS cohort in a comprehensive and unbiased manner , may be central players in aberrant processes associated with sarcoidosis , 43. In d manner . The resMonster analysis yielded a transcription factor to target gene network consisting of 106,029 edges in sarcoidosis cases and a corresponding network with differing strengths of interaction (edge weights) in controls from 81 transcription factors identified within 1,379 total genes in MAM. Examination of DTFI in MAM identified 11 transcription factors (13.58%) with significantly different targeting patterns in sarcoidosis compared to controls (FDR < 0.1)  and in the UCSF (24.48%) cohorts defined the “low” lymphocyte groups. Conversely, CD14+ monocytes greater than or equal to 31.96% and 31.84% median percentages defined the “high” monocyte groups in the ACCESS and UCSF cohorts, respectively. Surprisingly, the LSM derived calibration model pertaining to transcription factors ARID1B, CDK2, CRY1, and KMT2A resulted in 5 misclassifications and <70% predictive accuracy for both cell subsets; while all other calibration models were capable of predicting subjects with low CD4+ T-cells and high CD14+ monocytes accurately and were considered for validation  in a large cohort of healthy adults that was inclusive of African American and Caucasian subjects and corroborated reduced CD4+ T-cell proportions in all subjects within the “low” group  and CD4+ T-cell proportions of PBMCs was ascertained. Imputed and clinical cell proportions in the UIC STAR cohort were highly and significantly correlated and demonstrated coefficients  in the CD4+ T-cell model and 37 in the CD14+/total monocyte model. Similarly, 40/87 and 34/87 target genes had high VIP scores in the corresponding CBX8 models. These genes represented the most influential determinants of PBMC proportions and likely comprise a core gene signature of sarcoidosis immune dysregulation , 61–63. Aberrant systemic T-cell activity is crucial in the immunopathogenesis of sarcoidosis and peripheral lymphopenia is of clinical significance , 68, 69.By analyzing gene expression regulation, we sought to discern disrupted regulatory pathways driving changes in lymphocyte activity and survival. Traditionally proposed mechanisms of peripheral lymphopenia uphold that it is a consequence of T-cell sequestration, infiltrated bone marrow, or impaired survival and function of regulatory T-cells , 13, 14.We characterized two significant and preserved gene co-expression modules of sarcoidosis and the transcription factors that influence immune network dysregulation related to PBMC function and survival. The best predictor of low CD4+ T-cell abundance was not LSM, which included lymphocyte activity genes, such as CD28, CD40LG, and LEF1, necessary for T-cell effector function, but MAM. Within MAM, aberrancies inciting transcriptional up-regulation of monocyte activity and cell death related processes may be instrumental to the development of paradoxical peripheral lymphopenia. Specifically, significantly altered regulatory activity of transcription factors CBX8, ETV6, SPI1, TLE3, and XRCC3 was central to the cell death subnetwork. The gene targets of ETV6, SPI1, and XRCC3 did not satisfy our criteria to consider these transcription factors significantly impactful as master regulators on circulating lymphocyte and monocyte proportions. However, it is worth noting that these transcription factors have established roles in DNA damage repair, cell proliferation, cell differentiation, and TNF and interferon inflammatory pathways in malignancies and infections previously associated with sarcoidosis , 70–80. CBX8 and TLE3 were determined to be master regulators within the MAM cell death subnetwork. Unlike the target genes of XRCC3, ETV6, and SPI1, the target genes of CBX8 and TLE3 were predictive of CD4+ T-cell and CD14+ monocyte proportions in the UCSF cohort and their predictive ability was substantiated on the UIC STAR cohort. CBX8, a polycomb protein, is involved in DNA damage repair and has been linked to cell proliferation and differentiation through epigenetic modifications , 82. Intvia different mechanisms. Consequently, aberrant expression of distinct key determinant genes related to specific phenotypes adversely impact monocyte-lymphocyte interaction pathways and contribute to maladaptive peripheral inflammatory signaling which in turn is associated with development of lymphopenia and sarcoidosis progression.Our findings in the MAM cell death subnetwork indicate that immune cell imbalances are significantly related to an aberrantly expressed core gene signature modulated by altered CBX8 and TLE3 regulatory activity. Within this core signature, preserved differential over-expression was found between the ACCESS cohort and 14 genes in the UCSF cohort and between the ACCESS cohort and 31 genes in the UIC STAR cohort. These genes exhibited high discriminatory capacity and were considered the most influential predictors and key determinants of CD14+/total monocyte expansion and CD4+ T-cell depletion. Despite accurate prediction of cell proportions in both the UCSF and UIC STAR cohorts, little overlap was observed between key determinants. This finding was not surprising given the variability that is observed among differential expression gene signatures in sarcoidosis and the fact that our predictive models were calibrated on treatment naïve subjects early in the course of their disease from the ACCESS cohort and validated on demographically and clinically heterogenous cohorts , 28–34. in-silico methodologies to determine transcription factor involvement and impute immune cell populations and despite the congruency observed on cell proportions from clinical tests in the UIC STAR cohort in-vitro or in-vivo validation will still be required in future studies to further substantiate these findings. With respect to the in-silico methods, Monster and Cibersort, we note that both make use of input datasets to derive associations with transcription factors and cell types, and neither dataset was developed on sarcoidosis patients. Lastly, other limitations arise from the depth and accuracy of information stored in the databases that were utilized to generate the input datasets and determine functional annotation which can be addressed by re-analyzing the data in the future as information within public databases is expanded with information derived from more advanced technologies.In this study, we leveraged gene expression data from various high-throughput “-omic” platforms and clinically heterogenous cohorts to determine an immune regulatory network and gene signature associated with lymphopenia in sarcoidosis and provide a methodology to incorporate data from multiple studies to improve our understanding of the immunopathogenesis of sarcoidosis. Even though there are advantages to utilizing a well-defined, matched, and treatment naïve case-control cohort of sarcoidosis with mixed phenotypes, we recognize that there are several limitations to our study that will need to be addressed in the future. As noted, by utilizing the ACCESS cohort as a reference data set, our study was possibly limited in its ability to identify pertinent findings that may be held in other gene modules such as the darkred module that demonstrated a trend towards an association with smoking and the red module that may represent a distinctly preserved signature of immune dysregulation in sarcoidosis that is influenced by differences in race, treatment, or other features that contributed to cohort heterogeneity. A larger study utilizing a different analytical approach or different parameters to contrast specific sarcoidosis phenotypes with other granulomatous and lymphopenic diseases, in addition to health, might have been better able to target transcription factors and pathways of interest in sarcoidosis and increase the statistical power and specificity of our findings. Additionally, though our findings on the UIC STAR cohort suggest that our gene signature is applicable to African Americans with sarcoidosis, our findings are likely to be more representative of immune dysregulation in Caucasians with sarcoidosis. It is crucial that subsequent studies focus on African Americans to better elucidate specific mechanisms underlying disease pathogenesis unique to this population and equitably address disparities. Furthermore, this study utilized data obtained from different “bulk” transcriptomic methodologies and demonstrated preservation of findings; however, it should also be noted that despite efforts to rigorously normalize data and utilize statistics that are agnostic to type of “-omic” platform utilized (such as the Z-summary statistic and PLS-DA models) the possibility that gene selection bias was introduced into our signature by a specific platform cannot be fully discounted. By utilizing “bulk” transcriptomic methodologies our study was also limited in its ability to directly assess involvement of specific PBMC subsets. To address this, in-depth single-cell transcriptomic analyses utilizing the same methodology could be employed to provide a comprehensive assessment of cell-specific dysregulation. Our analytical strategy also relied on In conclusion, we determined master regulator transcription factors and target genes of immune dysregulation that are linked to characteristic paradoxical lymphopenia in sarcoidosis. Notably, we identified that differential transcriptional activity in monocytes was the best predictor of low lymphocyte counts. Crosstalk between monocytes and lymphocytes possibly involving the TNF pathway and Fc-γ receptors, as well as other features of innate immunity were the most likely dysregulated pathways involved with lymphopenia. Previous studies have established roles for both monocyte hyperactivity and T-lymphocyte dysfunction in sarcoidosis pathogenesis, but those analyses were limited in their assessment of the functional relationship between monocytes and lymphocytes , 68, 124The datasets presented in this study can be found in online repositories and the Supplementary Material. The names of the repository/repositories and accession numbers can be found in the article/This study was reviewed and approved by the University of Illinois Institutional Review Board (Protocol #s: 2019-0452 and 2016-0063). Written and informed consent was not required for protocol 2019-0452 in accordance with the national legislation and the institutional requirements. Written and informed consent was obtained from all subjects who participated in protocol 2016-0063 in accordance with the national legislation and the institutional requirements.A Case Control Etiologic Study of Sarcoidosis (ACCESS) Research Group Clinical Centers:Beth Israel Deaconess Medical Center: Steven E. Weinberger, M.D.; Patricia Finn, M.D.; Erik Garpestad, M.D.; Allison Moran, R.N.Georgetown University Medical Center: Henry Yeager, Jr., M.D.; David L. Rabin, M.D.; Susan Stein, M.A.Case Western Reserve University - Henry Ford Health Sciences Center: Michael C. Iannuzzi, M.D.; Benjamin A. Rybicki, Ph.D.; Marcie Major, R.N.; Mary Maliarik, Ph.D.; John Popovich, Jr., M.D.Johns Hopkins University School of Medicine: David R. Moller, M.D.; Carol J. Johns, M.D.*; Cynthia Rand, Ph.D.; Joanne Steimel, R.N.Medical University of South Carolina: Marc A. Judson, M.D.; Susan D’Alessandro, R.N., Nancy Heister, R.N.; Theresa Johnson, R.N.; Daniel T. Lackland, Dr.P.H.; Janardan Pandey, Ph.D.; Steven Sahn, M.D.; Charlie Strange, M.D.Mount Sinai Medical Center: Alvin S. Teirstein, M.D.; Louis DePalo, M.D.; Sheldon Brown, M.D.; Marvin Lesser, M.D.; Maria L. Padilla, M.D.; Marilyn MarshallNational Jewish Medical and Research Center: Lee S. Newman, M.D., M.A.; Cecile Rose,M.D., M.P.H.; Juliana Barnard, M.A.University of Cincinnati Medical Center: Robert P. Baughman, M.D.; Elyse E. Lower, M.D.; Donna B. WingetUniversity of Iowa College of Medicine: Geoffrey McLennan, M.D., Ph.D.; Gary Hunninghake, M.D.; Chuck Dayton, B.S.Pharm.; Linda Powers, M.S.University of Pennsylvania and Medical College of Pennsylvania - HahnemannMedical Centers: Milton D. Rossman, M.D.; Eddy A. Bresnitz, M.D.; Ronald Daniele, M.D., Jackie Regovich, M.P.H.; William Sexauer, M.D.National Heart, Lung, and Blood Institute: Robert Musson, Ph.D.; Joanne Deshler; Paul Sorlie, Ph.D.; Margaret Wu, Ph.D.Study Chairman: Reuben Cherniack, M.D.Study Co-Chairman: Lee Newman, M.D.Clinical Coordinating CenterClinical Trials & Surveys Corp.: Genell L. Knatterud, Ph.D., Michael L. Terrin, M.D., Bruce W. Thompson, Ph.D., Kathleen Brown, Ph.D., Margaret Frederick, Ph.D., Frances LoPresti, M.S., Patricia Wilkins, B.S., Martha Canner, M.S., Judy DotsonCentral Repository:McKesson Bioservices : Steve Lindenfelser BBI-Biotech Research Laboratories : Mark Cosentino, Ph.D.* Deceased.Central Laboratories:DNA Core Laboratory: Mary Maliarik, Ph.D.BAL Central Laboratory: Robert Baughman, M.D.HLA Class II Typing Laboratory: Milton Rossman, M.D.; Dimitri Monos, Ph.D.; Chung Wha Lee, Ph.D., Boyana Cizman, Ph.D.Etiologic Antigen in Kveim Reagent Laboratory: David Moller, M.D.Immunogenetics Laboratory: Janardan Pandey, Ph.D.L-Forms Core Laboratory: Peter Almenoff, M.D.; Ian Brett, Sheldon Brown, M.D.; Marvin Lesser, M.D.Pathogenic T Cells Laboratory: Lee Newman, M.D.; Brian Kotzin, M.D.Ribosomal DNA Core Laboratory: Geoffrey McLennan, M.D., Ph.D., Gary Hunninghake, M.D.RNA Core Laboratory: Patricia Finn, M.D.Random Digit Dialing Interview Group:Telesurveys Research Associates: Richard D. Jaffe, M.A.Executive Committee:Reuben Cherniack, M.D. (Chair)Robert P. Baughman, M.D. (9/1/98-8/31/99)Joanne DeshlerMichael C. Iannuzzi, M.D.; Marc A. Judson, M.D. (9/1/96-8/31/97); 9/1/00-6/30/01)Genell L. Knatterud, Ph.D.Geoffrey McLennan, M.D. (9/1/97-8/31/98)David R. Moller, M.D. Robert A Musson, Ph.D.Lee S. Newman, M.D.Milton D. Rossman, M.D. Alvin S. Teirstein, M.D. (9/1/97-8/31/98)Michael L. Terrin, M.D. M.P.H.Steven E. Weinberger, M.D. (9/1/97-3/31/98)Henry Yeager, Jr., M.D. (9/1/98-8/31/99)Data Safety and Monitoring Board:William Martin, M.D. (Chair)Takamaru Ashikaga, Ph.D.David B. Coultas, M.D.Gerald S. Davis, M.D.Fred Gifford, Ph.D.James J. Schlesselman, Ph.D.Diane Stover, M.D.Ex Officio:Reuben Cherniack, M.D., Genell L. Knatterud, Ph.D., Robert Musson, Ph.D.Lee Newman, M.D.CA, YH, CS and BT conceived, designed experiments, and performed experiments. Specimens and clinical data were collected by the ACCESS Research Group. CA, YH, and NE performed ACCESS clinical database abstraction. WW indexed, annotated and quantified transcriptomic libraries. CA, CAS, YH and BT analyzed data. DP, PF contributed reagents and materials. CA, CAS, YH, NS, DP and PF wrote the manuscript. All authors contributed to the article and approved the submitted version.Support for this study was provided by NIH grant numbers R01 HL138628-01A1S1, T32 HL082547, and F30 HL137267 for the University of Illinois at Chicago Division of Pulmonary, Critical Care, Sleep and Allergy.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.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.This manuscript was prepared using A Case Controlled Etiologic Study of Sarcoidosis (ACCESS) Research Materials obtained from the NHLBI Biologic Specimen and Data Repository Information Coordinating center and does not necessarily reflect the opinions or views of the ACCESS or the NHLBI.