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About 85% of patients with colorectal cancer (CRC) have the non-microsatellite instability-high (non-MSI-H) subtype, and many cannot benefit from immune checkpoint blockade. A potential reason for this is that most non-MSI-H colorectal cancers are immunologically "cold" due to poor CD8

Iron overload can contribute to the progression of nonalcoholic fatty liver disease (NAFLD) to nonalcoholic steatohepatitis (NASH). Hepcidin (Hamp), which is primarily synthesized in hepatocytes, is a key regulator of iron metabolism. However, the role of Hamp in NASH remains unclear. Therefore, we aimed to elucidate the role of Hamp in the pathophysiology of NASH.Male mice were fed a choline-deficient L-amino acid-defined (CDAA) diet for 16 weeks to establish the mouse NASH model. A choline-supplemented amino acid-defined (CSAA) diet was used as the control diet. Recombinant adeno-associated virus genome 2 serotype 8 vector expressing Hamp (rAAV2/8-Hamp) or its negative control (rAAV2/8-NC) was administered intravenously at week 8 of either the CDAA or CSAA diet.rAAV2/8-Hamp treatment markedly decreased liver weight and improved hepatic steatosis in the CDAA-fed mice, accompanied by changes in lipogenesis-related genes and adiponectin expression. Compared with the control group, rAAV2/8-Hamp therapy attenuated liver damage, with mice exhibiting reduced histological NAFLD inflammation and fibrosis, as well as lower levels of liver enzymes. Moreover, α-smooth muscle actin-positive activated hepatic stellate cells (HSCs) and CD68-postive macrophages increased in number in the CDAA-fed mice, which was reversed by rAAV2/8-Hamp treatment. Consistent with theOverexpression of Hamp using rAAV2/8-Hamp robustly attenuated liver steatohepatitis, inflammation, and fibrosis in an animal model of NASH, suggesting a potential therapeutic role for Hamp.

Tryptophan 2,3-dioxygnease 2 (TDO2) is specific for metabolizing tryptophan to kynurenine (KYN), which plays a critical role in mediating immune escape of cancer. Although accumulating evidence demonstrates that TDO2 overexpression is implicated in the development and progression of multiple cancers, its tumor-promoting role in esophageal squamous cell carcinoma (ESCC) remains unclear. Here, we observed that TDO2 was overexpressed in ESCC tissues and correlated significantly with lymph node metastasis, advanced clinical stage, and unfavorable prognosis. Functional experiments showed that TDO2 promoted tumor cell proliferation, migration, and colony formation, which could be prevented by inhibition of TDO2 and aryl hydrocarbon receptor (AHR). Further experimentation demonstrated that TDO2 could promote the tumor growth of KYSE150 tumor-bearing model, tumor burden of C57BL/6 mice with ESCC induced by 4-NQO, enhance the expression of phosphorylated AKT, with subsequent phosphorylation of GSK3β, and polarization of M2 macrophages by upregulating interleukin-8 (IL-8) to accelerate tumor progression in the tumor microenvironment (TME). Collectively, our results discovered that TDO2 could upregulate IL-8 through AKT/GSK3β to direct the polarization of M2 macrophages in ESCC, and suggested that TDO2 could represent as an attractive therapeutic target and prognostic marker to ESCC., Graphical abstract TDO2 expressed in ESCC cells participates in the regulation of the immune response and constitutes potential target for immunotherapy.Image 1

Continuous wavelet transform (CWT) has been proven to be an effective tool in feature extraction of non-stationary bio-signals. Therefore, hardware implementation of CWT has been widely investigated in wearable sensor integrated with local intelligence algorithm. To realize the feature extraction from low-frequency bio-signals, the design method of low-frequency Gm-C wavelet filter used in wearable sensor has been proposed in this paper. To alleviate the power constraint by wearable device, the Leap-Frog multiple-loop feedback filter structure is employed, which has low circuit complexity and sensitivity. Also, the transconductor consisting of simple differential pair is employed as Gm cell. By using low level bias current and deep weak inversion, low transconductance can be achieved to realize low frequency operation. A sixth-order Gaussian wavelet filter is designed in 0.18 $$\upmu$$ m CMOS process. Simulation results show that power consumption is only 392 pW at center frequency of 5.9 Hz, corresponding to dynamic range of 42.7 dB and figure-of-merit of 2.59 $$\times$$ 10 $$^{-13}$$ . Experiment result shows that the proposed wavelet filter can be used for accurate extraction of transient features in biomedical signal processing.

© 2021 John Wiley & Sons, Ltd. This is the accepted manuscript version of an article which has been published in final form at https://doi.org/10.1002/cta.3032

As an emerging immune checkpoint, CD73 has received more attention in the past decade. Inhibition of CD73 enzymatic activity can enhance antitumor immunity. Several CD73 inhibitors have been identified by in vitro assays in recent years, but they remain premature for clinical application, indicating that more novel CD73 inhibitors should be studied. Herein, we aimed to identify novel CD73 inhibitors that hopefully are suitable drug candidates by using computer-aided drug discovery and enzymatic-based assays. Five-hundred molecules with high binding affinity were retrieved from the Chemdiv-Plus database by using a structure-based virtual screening approach. Then, we analyzed the drug properties of these molecules and obtained 68 small molecules based on the oral noncentral nervous system (CNS) drug profile. The inhibition rates of these molecules against CD73 enzymatic activities were determined at a concentration of 100 μM, and 20 molecules had an inhibition rate greater than 20%, eight of which were dose-dependent, with IC50 values of 6.72-172.1 μM. Among the screening hits, phelligridin-based compounds had the best experimental inhibition values. Modeling studies indicate that the phelligridin group is sandwiched by the rings of F417 and F500 residues. The identified inhibitors have a molecular weight of approximately 500 Dal and are predicted to form primarily hydrogen bonds with CD73 in addition to hydrophobic stacking interactions. In conclusion, novel inhibitors with satisfactory drug properties may serve as lead compounds for the development of CD73-targeting drugs, and the binding modes may provide insight for phelligridin-based drug design.

Wearable electroencephalography sensor integrated with seizure detection algorithm has become an emerging research topic in health management for patients with refractory epilepsy. To enhance seizure detection accuracy and reduce power consumption, this paper has proposed a selection method of optimal features customized for each patient. A feature pool consisting of several popular features is constructed. Then, the customized multi-domain features for each patient are selected from feature pool based on minimum redundancy maximum relevance. By removing redundant features, the computational complexity of on-line seizure detection can be greatly reduced, which is desired in wearable device where low-power and real-time operation are required. Four test datasets are used to evaluate the performance of proposed method, with detection accuracy calculated. Experiment results show that different datasets vary in optimal features. By using the proposed method, the detection accuracy and the value of area under receiver operating characteristic curve can be improved by 11.49% and 0.13 respectively, while feature dimension being reduced to 1.

Background Inhibitors targeting immune checkpoint were proved effective in cancer immunotherapy, such as PD-1/PD-L1 blockade. The novel immune checkpoint TIGIT/PVR plays critical roles in suppressing the anti-tumor effects of CD8+ T and NK cells, and dual blockade of TIGIT/PVR and PD-1/PD-L1 by antibody can elicit synergistic effects in tumor models and clinical trials. However, small molecules for TIGIT/PVR blockade have not been investigated. Methods The expression of PVR in tumors were analyzed by using TCGA, Oncomine and GEO database, and in cancer cell lines examined by flow cytometry. Natural product compounds were docked to PVR for virtual screening by using the software Molecular Operating Environment (MOE). Candidate compounds were further tested by biolayer interferometry-based binding assay, microscale thermophoresis assay and cell based blocking assay. The in vitro activity of the candidate compound was determined by MTT, peripheral blood mononuclear cells (PBMCs) activation assay and coculture assay. The anti-tumor effects and mechanism were also investigated by using MC38 tumor-bearing mice model and immune cell depletion tumor model. Results PVR was over-expressed in many tumor tissues and cancer cell lines, making it a promising therapeutic target. Through virtual screening, binding, and blocking assay, liothyronine was discovered to bind PVR and block the interaction of TIGIT/PVR. Liothyronine could enhance the function of CD4+ and CD8+ T cells in PBMCs. Besides, in the Jurkat-hTIGIT and CHOK1-hPVR coculture assay, liothyronine could reverse the IL-2 secretion inhibition resulted by TIGIT/PVR ligation. Although had no influence on the proliferation of tumor cells in vitro, liothyronine could significantly inhibit tumor growth when administrated in vivo, by enhancing CD8+ T cell infiltration and immune responses in the tumor bearing mice. The immune cell depletion model showed that the anti-tumor effects of liothyronine depends on CD4+ T cells, CD8+ T cells and NK cells. Conclusions A small molecule liothyronine was discovered to serve as a potential candidate for cancer immunotherapy by blocking the immune checkpoint TIGIT/PVR. Graphical abstract

Introduction: An outbreak of coronavirus disease 2019 (COVID-19) occurred in Wuhan, Hubei Province, China. This study aimed to analyze the clinical and epidemiologic characteristics of patients with COVID-19 to better differentiate the suspected patients in Beijing, China. Methodology: This was a retrospective, single-center study. Clinical and epidemiologic data were collected from suspected patients with COVID-19 admitted to Beijing Ditan Hospital from January 29 to February 21, 2020. Results: One hundred and six patients (60 males and 46 females, median age 36 years) were enrolled. Thirty-six patients were ultimately laboratory confirmed. Fifty-three were excluded from the diagnosis of COVID-19. The remaining 17 patients were highly suspected, although their nucleic acid tests were repeatedly negative. The confirmed patients and highly suspected patients had a significantly higher proportion of epidemiologic history than the excluded patients (P < 0.001). There was no significant difference in clinical symptoms or the underlying diseases among the three groups. The confirmed patients had a higher frequency of lymphopenia and eosinopenia than the highly suspected and excluded patients. Chest computed tomography scans showed bilateral lung involvement, and ground-glass opacity was more likely observed in the confirmed patients. Conclusion: The clinical features of the confirmed patients with COVID-19 were insufficient for early diagnosis of COVID-19. The epidemiologic history was of great significance in the early diagnosis of COVID-19. More sensitive diagnostic methods are needed to aid the differential diagnosis of suspected patients with COVID-19.

The low response rate and adaptive resistance of PD-1/PD-L1 blockade demands the studies on novel therapeutic targets for cancer immunotherapy. We discovered that a novel immune checkpoint TIGIT expressed higher than PD-1 in many tumors especially anti-PD-1 resistant tumors. Here, mirror-image phage display bio-panning was performed using the d-enantiomer of TIGIT synthesized by hydrazide-based native chemical ligation. d-peptide D TBP-3 was identified, which could occupy the binding interface and effectively block the interaction of TIGIT with its ligand PVR. D TBP-3 showed proteolytic resistance, tumor tissue penetrating ability, and significant tumor suppressing effects in a CD8+ T cell dependent manner. More importantly, D TBP-3 could inhibit tumor growth and metastasis in anti-PD-1 resistant tumor model. This is the first d-peptide targeting TIGIT, which could serve as a potential candidate for cancer immunotherapy.

COVID-19 (coronavirus disease 2019) is an infectious disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and discovered in 2019. The clinical manifestations include fever, coughing, difficulty in breathing and even death from multiple organ failure. Nucleic acid test is the golden standard method for confirmation of infection. According to the Chinese ‘Seventh Edition of the COVID-19 Diagnosis and Treatment Protocol’, suspected patients with negative nucleic acid tests from two consecutive airway specimens can be excluded from diagnosis and released from quarantine. The current report describes a suspected COVID-19 case that had a history of close contact with a COVID-19 patient. The diagnosis was confirmed after the SARS-CoV-2 nucleic acid was detected after four sputum sample tests (sampling interval of at least 24 h).

As the evolution of traditional electroencephalogram (EEG) monitoring unit for epilepsy diagnosis, wearable ambulatory EEG (WAEEG) system transmits EEG data wirelessly, and can be made miniaturized, discrete and social acceptable. To prolong the battery lifetime, analog wavelet filter is used for epileptic event detection in WAEEG system to achieve on-line data reduction. For mapping continuous wavelet transform to analog filter implementation with low-power consumption and high approximation accuracy, this paper proposes a novel approximation method to construct the wavelet base in analog domain, in which the approximation process in frequency domain is considered as an optimization problem by building a mathematical model with only one term in the numerator. The hybrid genetic algorithm consisting of genetic algorithm and quasi-Newton method is employed to find the globally optimum solution, taking required stability into account. Experiment results show that the proposed method can give a stable analog wavelet base with simple structure and higher approximation accuracy compared with existing method, leading to a better spike detection accuracy. The fourth-order Marr wavelet filter is designed as an example using Gm-C filter structure based on LC ladder simulation, whose power consumption is only 33.4 pW at 2.1Hz. Simulation results show that the design method can be used to facilitate low power and small volume implementation of on-line epileptic event detector.

Although the blockade of immune checkpoint PD-1/PD-L1 has achieved great success, the lack of tumor-infiltrating immune cells and PD-L1 expression in the tumor microenvironment results in a limited response in certain tumor types. Thus, rational and optimal combination strategies were urgently needed. The combination of PD-1/PD-L1 blockade and anti-angiogenic therapy has been reported to have great potential. Here, a chimeric peptide OGS was designed by conjugating the peptides OPBP-1 (8-12) and

The antioxidant transcription factor NFE2L1 (also called Nrf1) acts as a core regulator of redox signaling and metabolism homeostasis, and thus, its dysfunction results in multiple systemic metabolic diseases. However, the molecular mechanism(s) by which NFE2L1 regulates glycose and lipid metabolism remains elusive. Here, we found that loss of NFE2L1 in human HepG2 cells led to a lethal phenotype upon glucose deprivation and NFE2L1 deficiency could affect the uptake of glucose. Further experiments revealed that glycosylation of NFE2L1 enabled it to sense the energy state. These results indicated that NFE2L1 can serve as a dual sensor and regulator of glucose homeostasis. The transcriptome, metabolome, and seahorse data further revealed that disruption of NFE2L1 could reprogram glucose metabolism to aggravate the Warburg effect in NFE2L1-silenced hepatoma cells, concomitant with mitochondrial damage. Co-expression and Co-immunoprecipitation experiments demonstrated that NFE2L1 could directly interact and inhibit AMPK. Collectively, NFE2L1 functioned as an energy sensor and negatively regulated AMPK signaling through directly interacting with AMPK. The novel NFE2L1/AMPK signaling pathway delineate the mechanism underlying of NFE2L1-related metabolic diseases and highlight the crosstalk between redox homeostasis and metabolism homeostasis.

NFE2L1 (also called Nrf1) acts a core regulator of redox signaling and metabolism homeostasis, and thus its dysfunction results in multiple systemic metabolic diseases. However, the molecular mechanism(s) by which NFE2L1 regulates glycose and lipid metabolism is still elusive. Here, we found that the loss of NFE2L1 in human HepG2 cells led to a lethal phenotype upon glucose deprivation. The uptake of glucose was also affected by NFE2L1 deficiency. Further experiments unveiled that although the glycosylation of NFE2L1 was monitored through the glycolysis pathway, it enabled to sense the energy state and directly interacted with AMPK. These indicate that NFE2L1 can serve as a dual sensor and regulator of glucose homeostasis. In-depth sights into transcriptome, metabolome and seahorse data further unraveled that glucose metabolism was reprogrammed by disruption of NFE2L1, so as to aggravate the Warburg effect in NFE2L1-silenced hepatoma cells, along with the mitochondrial damage observed under the electron microscope. Collectively, these demonstrate that disfunction of NFE2L1 triggers the uncontrollable signaling by AMPK towards glucose metabolism reprogramming in the liver cancer development.

BackgroundThe development of cancer is largely dependent on the accumulation of somatic mutations, indicating the potential to develop cancer chemoprevention agents targeting mutation drivers. However, ideal cancer chemoprevention agents that can effectively inhibit the mutation drivers have not been identified yet.MethodsThe somatic mutation signatures and expression analyses of APOBEC3B were performed in patient with pan-cancer. The computer-aided screening and skeleton-based searching were performed to identify natural products that can inhibit the activity of APOBEC3B. 4-nitroquinoline-1-oxide (4-NQO)-induced spontaneous esophageal squamous cell carcinoma (ESCC) and azoxymethane/dextran sulfate sodium (AOM/DSS)-induced spontaneous colon cancer mouse models were conducted to investigate the influences of APOBEC3B inhibitor on the prevention of somatic mutation accumulation and cancer progression.ResultsHere, we discovered that the cytidine deaminase APOBEC3B correlated somatic mutations were widely observed in a variety of cancers, and its overexpression indicated poor survival. SMC247 (3, 5-diiodotyrosine), as a source of kelp iodine without side effects, could strongly bind APOBEC3B (KD=65 nM) and effectively inhibit its deaminase activity (IC50=1.69 µM). Interestingly, 3, 5-diiodotyrosine could significantly reduce the clusters of mutations, prevent the precancerous lesion progression, and prolong the survival in 4-NQO-induced spontaneous ESCC and AOM/DSS-induced spontaneous colon cancer mouse models. Furthermore, 3, 5-diiodotyrosine could reduce colitis, increase the proportion and function of T lymphocytes via IL-15 in tumor microenvironment. The synergistic cancer prevention effects were observed when 3, 5-diiodotyrosine combined with PD-1/PD-L1 blockade.ConclusionsThis is the first prove-of-concept study to elucidate that the natural product 3, 5-diiodotyrosine could prevent somatic mutation accumulation and cancer progression through inhibiting the enzymatic activity of APOBEC3B. In addition, 3, 5-diiodotyrosine could reduce the colitis and increase the infiltration and function of T lymphocytes via IL-15 in tumor microenvironment. 3, 5-diiodotyrosine combined with PD-1/PD-L1 blockade could elicit synergistic cancer prevention effects, indicating a novel strategy for both prevent the somatic mutation accumulation and the immune-suppressive microenvironment exacerbation.

Since block copolymers are able to self-assemble into various polymeric architectures, it is intriguing to explore a unique self-assembly strategy for polymers. Two different metallic oxides [manganese dioxide (MnO2) and zinc oxide (ZnO)] are displayed herein to demonstrate this self-assembly mechanism of polymers. In situ generation of metallic oxides induces self-assembly of block copolymers to form polymeric hybrid micelles with tunable stability in aqueous solutions. These final ZnO-cross-linked polymeric micelles exhibited a high drug loading capacity of 0.41 mg mg-1 toward doxorubicin (DOX), whereas DOX-loaded ZnO-cross-linked polymeric micelles could be broken down into Zn2+ and polymer scraps, which facilitated drug release in tumor microenvironments. Both in vitro and in vivo investigations showed that the drug-loaded ZnO-cross-linked polymeric micelles effectively suppressed tumor growth. Accordingly, the present study demonstrates a novel strategy of polymer self-assembly for fabricating polymeric architectures that can potentially provide insight for developing other polymeric architectures.

Colorectal cancer is one of the deadliest cancers, and the discovery of new diagnostic biomarkers and therapeutic targets is vital. Interleukin-36α (IL-36α) is a proinflammatory factor that can initiate the inflammatory response and promote the systemic T helper-1 (Th1) immune response. In this study, we investigated the immunological role of IL-36α in CRC. We found that IL-36α was downregulated in human CRC tissues. Patients with high IL-36α levels showed better survival and low IL-36α expression was significantly associated with greater tumor distal metastasis and TNM stage. We constructed two cell lines overexpressing IL-36α (CT26-IL-36α and HT29-IL-36α cells). In vitro assays revealed that IL-36α overexpression reduced the proliferation, migration, and invasion of CT26-IL-36α, and HT29-IL-36α cells. Using CT26-vector and CT26-IL-36α tumor mouse model and lung metastasis models, we found that IL-36α overexpression elicited a significant antitumor effect and inhibited lung metastasis in vivo. These inhibitory effects were associated with an increase in the number of CD3

Cardiac arrhythmias are the most common cause of sudden cardiac death worldwide. Lengthening the ventricular action potential duration (APD) either congenitally or via pathologic or pharmacologic means, predisposes to a life-threatening ventricular arrhythmia, Torsade de Pointes. IKs, a slowly activating K+ current plays a role in action potential repolarization. In this study, we screened a chemical library in silico by docking compounds to the voltage sensing domain (VSD) of the IKs channel. Here we show that C28 specifically shifted IKs VSD activation in ventricle to more negative voltages and reversed drug-induced lengthening of APD. At the same dosage, C28 did not cause significant changes of the normal APD in either ventricle or atrium. This study provides evidence in support of a computational prediction of IKs VSD activation as a potential therapeutic approach for all forms of APD prolongation. This outcome could expand the therapeutic efficacy of a myriad of currently approved drugs that may trigger arrhythmias.Significance statementC28, identified by in silico screening, specifically facilitated voltage dependent activation of a cardiac potassium ion channel, IKs. C28 reversed drug-induced prolongation of action potentials, but minimally affected the normal action potential at the same dosage. This outcome supports a computational prediction of modulating IKs activation as a potential therapy for all forms of action potential prolongation, and could expand therapeutic efficacy of many currently approved drugs that may trigger arrhythmias.

Strategies boosting both innate and adaptive immunity have great application prospects in cancer immunotherapy. Antibodies dual blocking the innate checkpoint CD47 and adaptive checkpoint PD-L1 or TIGIT could achieve durable anti-tumor effects. However, a small molecule dual blockade of CD47/SIRPα and TIGIT/PVR pathways has not been investigated. Here, an elevated expression of CD47 and PVR was observed in tumor tissues and cell lines analyzed with the GEO datasets and by flow cytometry, respectively. Compounds approved by the FDA were screened with the software MOE by docking to the potential binding pockets of SIRPα and PVR identified with the corresponding structural analysis. The candidate compounds were screened by blocking and MST binding assays. Azelnidipine was found to dual block CD47/SIRPα and TIGIT/PVR pathways by co-targeting SIRPα and PVR. In vitro, azelnidipine could enhance the macrophage phagocytosis when co-cultured with tumor cells. In vivo, azelnidipine alone or combined with irradiation could significantly inhibit the growth of MC38 tumors. Azelnidipine also significantly inhibits the growth of CT26 tumors, by enhancing the infiltration and function of CD8+ T cell in tumor and systematic immune response in the tumor-draining lymph node and spleen in a CD8+ T cell dependent manner. Our research suggests that the anti-hypertensive drug azelnidipine could be repositioned for cancer immunotherapy.

Background TIGIT, as a novel immune checkpoint molecule involved in T cell and NK cell anergy, could induce the immune tolerance and escape through binding with its ligand PVR. Blockade of TIGIT/PVR is considered as a promising strategy in cancer immunotherapy. However, to facilitate the design of inhibitors targeting TIGIT/PVR, the structural characteristics and binding mechanism still need to be further studied. Methods In this study, molecular dynamics (MD) simulations and in silico mutagenesis were used to analyze the interaction between TIGIT and its ligand PVR. Then, PVR mutants were designed and their activities were determined by using TIGIT overexpressed Jurkat cells. Results The results suggested that the loops of PVR (CC′ loop, C′C″ loop, and FG loop) underwent a large intra-molecular rearrangement, and more hydrogen bond crosslinking between PVR and TIGIT were formed during MD simulations. The potential residues for PVR to interact with TIGIT were identified and utilized to predict high affinity PVR mutants. Through the biological activity evaluation, four PVR mutants (PVRS72W, PVRS72R, PVRG131V and PVRS132Q) with enhanced affinity to TIGIT were discovered, which could elicit more potent inhibitory effects compared with the wild type PVR. Conclusions The MD simulations analysis provided new insights into the TIGIT/PVR interaction model, and the identified PVR mutants (PVRS72W, PVRS72R, PVRG131V and PVRS132Q) could serve as new candidates for immunotherapy to block TIGIT/PVR.

Purpose : Seizure detection algorithms (SDAs) based on electroencephalography (EEG) have been described in previous studies, but the imbalanced data distribution of ictal and interictal states continue to pose a technical challenge. This study proposes a novel algorithm to address the imbalanced classification problem and improve seizure detection performance. Method : The proposed algorithm is designed based on hybrid sampling and a cost-sensitive (CS) classifier. Hybrid sampling resamples the imbalanced EEG data at data-level, and the CS classifier, which is used as an algorithm-level tool, reduces the overall misclassification cost of seizure detection. The synthetic minority oversampling technique and undersampling TomekLink technique are combined to reduce the imbalanced ratio between ictal and interictal states while retaining the generalization ability. Finally, CS support vector machine classifies the resampled EEG feature vectors, assigning different cost sensitive parameters to moderate the poor performance resulting from the imbalanced distribution problem. Result : The proposed algorithm improved the average sensitivity and AUC by 46.67% and 0.0482, respectively, compared with the original results without using the imbalanced EEG data processing (IEDP) technique. Experimental results showed that an average sensitivity and AUC of 86.34% and 0.9837, respectively, could be obtained across all cases. Finally, a performance evaluation showed that the proposed algorithm outperformed published methods in terms seizure detection. Conclusion : A fusion algorithm combining data- and algorithm-level methods can achieve high sensitivity and AUC compared with existing IEDP methods. Thus, SDA performance can be improved, enabling their clinical use with EEG-based SMSs.

Additional file 1: Figure S1. Protein topology of hTIGIT and hPVR, along with the structure of TIGIT/PVR complex. (a, b) hTIGIT and hPVR contained ectodomain (blue), transmembrane domain (red) and intracellular domain (purple). The glycosylation sites of the extracellular domain were marked with a yellow ball. (c, d) The residue sequences of hTIGIT and hPVR, where the secondary structure α-helix and β-sheet were noted by cyan and yellow shadow respectively, and the residues forming the disulfide bond were represented by red. (e) Model of hTIGIT complexed with hPVR, which was retrieved from heterotetrameric hTIGIT/hPVR complex (PDB ID code 3UDW). The secondary structure and disulfide bond were shown in the same way as above. Figure S2. Stable cell lines of hPVR mutants. (a) CHOK1 cells overexpressing WT and mutant hPVR were stained with anti-human PVR APC. For each histogram, the filled blue histogram with blue line is the hPVR specific antibody and the histogram with red lines is the isotype control. (b) Lysates of CHOK1, CHOK1-hPVR, and CHOK1-mutants were used for western blotting. The blot was developed by chemiluminescence. Figure S3. Relationship between protein expression and binding affinity. Protein expression were normalized versus wild type hPVR cells. Graphs showed the correlation between protein expression and binding affinity When hTIGIT-Fc at different concentrations. The Pearson correlation coefficient and P were shown. Figure S4. Binding affinity of PVR mutants fused with EGFP to hTIGIT-His. The membrane protein of PVR mutants fused with EGFP was used to detect the binding affinity with hTIGIT-His. The concentration of TIGIT-His was serially diluted by two-fold with 0.000153 μM from 25 μM. The KD values of PVR mutants with hTIGIT-His were shown. Graphs were representative of three independent experiments..

Additional file 2: Table S1. Alanine scanning of important residue positions.