Disposable stochastic sensors obtained by nanolayer deposition of copper, graphene, and copper-graphene composite on silk for the determination of isocitrate dehydrogenases 1 and 2.

Three disposable stochastic sensors designed using nanolayer deposition of copper (Cu), graphene (GR), and copper-graphene (Cu-GR) composite on the silk textile, as substrate, were modified with chitosan (n=371-744), for biomedical analysis. Isocitrate dehydrogenase 1 (IDH1) and isocitrate dehydrogenase 2 (IDH2) served as model analytes for molecular recognition and quantification in biological samples such as whole blood and brain tumor tissue samples. The best sensitivities (3.77×107s µg mL-1 for IDH1, and 1.88×107s µg mL-1 for IDH2) and the lowest limits of quantification (10-2fg mL-1 for IDH1, and 5×10-2fg mL-1 for IDH2) for both IDH1 and IDH2 were recorded for the disposable stochastic sensors based on chitosan/graphene nanolayer. Very good correlations between the screening method based on disposable stochastic sensors and enzyme-linked immunosorbent assay (ELISA) were obtained; this was also proved by the results obtained using the paired t-test.

The role of deep learning-based survival model in improving survival prediction of patients with glioblastoma.

This retrospective study has been conducted to validate the performance of deep learning-based survival models in glioblastoma (GBM) patients alongside the Cox proportional hazards model (CoxPH) and the random survival forest (RSF). Furthermore, the effect of hyperparameters optimization methods on improving the prediction accuracy of deep learning-based survival models was investigated. Of the 305 cases, 260 GBM patients were included in our analysis based on the following criteria: demographic information (i.e., age, Karnofsky performance score, gender, and race), tumor characteristic (i.e., laterality and location), details of post-surgical treatment (i.e., time to initiate concurrent chemoradiation therapy, standard treatment, and radiotherapy techniques), and last follow-up time as well as the molecular markers (i.e., O-6-methylguanine methyltransferase and isocitrate dehydrogenase 1 status). Experimental results have demonstrated that age (Elderly > 65: hazard ratio [HR] = 1.63; 95% confidence interval [CI]: 1.213-2.18; p value = 0.001) and tumors located at multiple lobes ([HR] = 1.75; 95% [CI]: 1.177-2.61; p value = 0.006) were associated with poorer prognosis. In contrast, age (young < 40: [HR] = 0.57; 95% [CI]: 0.343-0.96; p value = 0.034) and type of radiotherapy (others include stereotactic and brachytherapy: [HR] = 0.5; 95%[CI]: 0.266-0.95; p value = 0.035) were significantly related to better prognosis. Furthermore, the proposed deep learning-based survival model (concordance index [c-index] = 0.823 configured by Bayesian hyperparameter optimization), outperformed the RSF (c-index = 0.728), and the CoxPH model (c-index = 0.713) in the training dataset. Our results show the ability of deep learning in learning a complex association of risk factors. Moreover, the remarkable performance of the deep-learning-based survival model could be promising to support decision-making systems in personalized medicine for patients with GBM.

Prognostic Biomarker ZNF311 and Its Correlation With Tumor Progression and Immune Infiltrates in Glioma.

BACKGROUND: Gliomas, particularly high-grade gliomas, are the most common primary brain tumors. From the Chinese Glioma Genome Atlas (CGGA) database, the relationships between the altered molecular pathways and gliomas could be easily observed. A close connection in the occurrence of the pathogenesis exists between the microenvironment, the glioma, and the associated genes. METHODS: Validation of the role of ZNF311 oncogene was confirmed by data from the CGGA dataset on glioblastoma and low-grade glioma. Furthermore, we used CIBERSORT to analyze the correlation between ZNF311 and cancer immune infiltrates. RESULTS: According to our analysis, ZNF311 was expressed higher in patients with grade-depended glioma with poor prognosis. In addition, we obtained valuable prognostic results between isocitrate dehydrogenase 1 (IDH1) and ZNF311 through the analysis of integrated correlations. Similarly, we simultaneously revealed the prognostic results between 1p/19q and ZNF311. In addition, we found that ZNF311 is correlated with a large number of tumor-infiltrating immune cells. CONCLUSIONS: Based on the study findings, we conclude that ZNF311 is potentially a novel biomarker for assessing prognosis and immune infiltration in glioblastoma and diffuse glioma cases.

High TRAF3IP3 Level Predicts Poor Prognosis of Patients with Gliomas.

BACKGROUND: Tumor necrosis factor receptor-related factor 3 (TRAF3) interacting protein 3 (TRAF3IP3) is involved in the development of immune tissues and the immune response of the body. Downregulated expression of TRAF3IP3 in malignant melanoma can inhibit tumor growth. The role of TRAF3IP3 in glioma is unknown. METHODS: We used the Wilcoxon rank sum test to compare the expression of TRAF3IP3 in glioma and normal tissues based on The Cancer Genome Atlas and Genotype Tissue Expression. Logistics regression was used to evaluate the relationship between TRAF3IP3 and clinicopathologic characters. Gene set enrichment analysis and single-sample gene set enrichment analysis were conducted to annotate biological function of TRAF3IP3. We used Kaplan-Meier and Cox regression to evaluate the prognostic value of TRAF3IP3. RESULTS: We downloaded RNA-seq data of 670 gliomas and 1157 normal tissues. TRAF3IP3 was highly expressed in gliomas (P < 0.001). High expression of TRAF3IP3 and higher World Health Organization grade (odds ratio [OR], 3.57 [2.42-5.34 CI]; P < 0.001), wild-type isocitrate dehydrogenase status (OR, 4.79 [3.40-6.83 CI]; P < 0.001), 1p/19q non-codeletion (OR, 15.32 [9.23-27.01 CI]; P < 0.001), mutant epidermal growth factor receptor status (OR, 2.77 [1.65-4.81 CI]; P < 0.001), worse histologic type (OR, 3.64 [2.48-5.43 CI]; P < 0.001) and worse primary therapy outcome (OR, 2.29 [1.47-3.61 CI]; P < 0.001) were significantly correlated. Six signaling pathways were significantly enriched in the TRAF3IP3 high-expression phenotype group, including JAK-STAT, interferon-γ, apoptosis, P53, programmed cell death protein 1, and CTLA-4 (cytotoxic T-lymphocyte-associated protein 4). High expression of TRAF3IP3 was associated with worse progression-free survival (hazard ratio [HR], 2.39 (1.39-3.01); P < 0.001), disease-free survival (HR, 3.02 (2.27-4.01); P < 0.001) and overall survival (HR, 2.87 (2.20-3.75); P < 0.001). CONCLUSIONS: TRAF3IP3 play an important role in the occurrence and development of glioma and may be a potential biomarker for the prognosis of glioma.

Using arterial-venous analysis to characterize cancer metabolic consumption in patients.

Understanding tumor metabolism holds the promise of new insights into cancer biology, diagnosis and treatment. To assess human cancer metabolism, here we report a method to collect intra-operative samples of blood from an artery directly upstream and a vein directly downstream of a brain tumor, as well as samples from dorsal pedal veins of the same patients. After performing targeted metabolomic analysis, we characterize the metabolites consumed and produced by gliomas in vivo by comparing the arterial supply and venous drainage. N-acetylornithine, D-glucose, putrescine, and L-acetylcarnitine are consumed in relatively large amounts by gliomas. Conversely, L-glutamine, agmatine, and uridine 5-monophosphate are produced in relatively large amounts by gliomas. Further we verify that D-2-hydroxyglutarate (D-2HG) is high in venous plasma from patients with isocitrate dehydrogenases1 (IDH1) mutations. Through these paired comparisons, we can exclude the interpatient variation that is present in plasma samples usually taken from the cubital vein.

Utility of isocitrate dehydrogenase 1 as a serum protein biomarker for the early detection of non-small-cell lung cancer: A multicenter in vitro diagnostic clinical trial.

We aimed to verify the expression status and diagnostic significance of isocitrate dehydrogenase 1 (IDH1) in non-small-cell lung cancer (NSCLC), especially during early stages. Serum IDH1 levels were measured by ELISA. A total of 1223 participants (660 patients with NSCLC, 276 healthy controls [HCs], 95 patients with benign pulmonary conditions [BPCs], 135 patients with other cancers [OCs], and 57 samples with interfering factors) were divided into a training cohort and a validation cohort according to 3 testing centers. The IDH1 concentrations in the NSCLC group were obviously higher than those in the control groups (P < .001). Area under the receiver operating characteristic curves (AUCs) for discriminating NSCLC patients from controls (HC, BPC, and OC) were 0.870 and 0.745 (sensitivity, 63.3% and 55.0%; specificity, 86.8% and 86.3%) in the training cohort and validation cohort, respectively. The AUCs for discriminating stage 0-IA lung cancer patients from HCs were 0.907 and 0.788 (sensitivity, 58.6% and 59.1%; specificity, 92.9% and 89.3%) in 2 cohorts, respectively. Isocitrate dehydrogenase 1 showed specificity for NSCLC and had no diagnostic value for other common cancers. Furthermore, IDH1 was significantly reduced in postoperative serum. Isocitrate dehydrogenase 1 shows clinical utility as a serum protein biomarker for the early diagnosis of NSCLC.

[Value of Serum Tumor Marker Isocitrate Dehydrogenase 1 in the Diagnosis of Lung Cancer].

Objective To investigate the clinical value of serum tumor marker isocitrate dehydrogenase 1(IDH1)in the diagnosis of lung cancer. Methods The general data were collected in lung cancer patients and non-lung cancer patients.The serum level of IDH1 was detected by enzyme-linked immunosorbent assay to evaluate its clinical significance in diagnosing lung cancer. Results The serum IDH1 level was significantly higher in lung cancer patients than in non-lung cancer patients [(7.12±6.98)ng/ml vs.(2.09±1.83)ng/ml,t=11.540,P<0.001].The serum IDH1 level in patients with adenocarcinoma or squamous cell carcinoma was significantly higher than that in patients with small cell lung cancer [(7.91±7.26)ng/ml vs.(2.76±2.27)ng/ml, t=6.345,P<0.001].The sensitivity of IDH1 in detecting lung cancer,stage Ⅰ/Ⅱ lung cancer,and stage Ⅲ/Ⅳ lung cancer was 47.4%,49.1%,and 46.3%,respectively. Conclusions Serum IDH1 has high sensitivities and specificities in the diagnosis and differential diagnosis of non-small cell lung cancer(squamous cell carcinoma and adenocarcinoma)and small cell lung cancer as well as the auxiliary diagnosis of stage Ⅰ and Ⅱ lung cancer.It is a valuable marker for the auxiliary diagnosis of lung cancer.

The Treatment of Gliomas in Adulthood.

BACKGROUND: Gliomas are the most common intrinsic tumors of the brain, with an incidence of 6 per 100 000 persons per year. Recent years have seen marked changes in the diagnosis and treatment of gliomas, with molecular parameters now being an integral part of the diagnostic evaluation. METHODS: This review is based on pertinent articles retrieved by a selective search in PubMed, with special attention to the new WHO glioma classification. RESULTS: The classification of gliomas on the basis of additional molecular parameters enables more accurate prognostication and serves as a basis for therapeutic decision-making and treatment according to precisely specified algorithms. PET scanning with 18F-fluoroethyl tyrosine and 11C-methionine for the measurement of metabolic activity in gliomas has further refined the diagnostic evaluation. The median overall survival of patients with glioblastoma who have undergone resection of all tumor tissue with a disrupted blood-brain barrier (i.e., all contrast-enhancing tumor tissue) has been prolonged to up to 20 months. The 5-year survival of patients with WHO grade II gliomas is now as high as 97% after near-total resection. The surgical resection of all contrast-enhancing tumor tissue and subsequent radiotherapy and chemotherapy remain the key elements of treatment. New surgical strategies and new methods of planning radiotherapy have made these techniques safer and more effective. The percutaneous application of tumor-treating fields is a new therapeutic option that has gained a degree of acceptance. Accompanying measures such as psycho-oncology and palliative care are very important for patients and should be considered mandatory. CONCLUSION: The consistent application of the existing multimodal treatment options for glioma has led in recent years to improved survival. Areas of important current and future scientific activity include immunotherapy and targeted and combined chemotherapy, as well as altered neurocognition, modern approaches to palliative care, and complementary therapies.

IDH2 is a novel diagnostic and prognostic serum biomarker for non-small-cell lung cancer.

Lung cancer is the most common leading cause of cancer-related death worldwide. Late diagnosis contributes to a high mortality rate and poor survival of this cancer. In our previous study, we found that IDH2 polymorphism rs11540478 is a risk factor for lung cancer. Here, we examined IDH2 protein expression in culture medium in which two non-small-cell lung cancer (NSCLC) cell lines, H460 and A549, were growing. We found that the IDH2 protein was elevated in the culture supernatant fraction in a time- and cell number-dependent manner. Next, we used ELISA methods to examine IDH2 protein level in serum from patients with NSCLC and healthy controls. We found that IDH2 protein levels in serum could distinguish NSCLC patients from healthy controls with an AUC (area under the curve) of 0.83 (95% confidence interval = 0.79-0.88). The IDH2 level was decreased in serum from NSCLC postsurgical patients compared with the paired presurgical serum. High serum IDH2 levels appear to correlate with poor survival in patients with NSCLC. These results suggest that IDH2 levels in the serum could be a new effective biomarker for the diagnosis and prognosis of NSCLC.

Prognostic and diagnostic potential of isocitrate dehydrogenase 1 in esophageal squamous cell carcinoma.

We aimed to investigate the pattern of expression and clinical significance of isocitrate dehydrogenase 1(IDH1) in esophageal squamous cell carcinoma (ESCC). The IDH1 expression was determined by quantitative real-time polymerase chain reaction, immunohistochemistry, and Western blot analysis using 38 pairs of frozen tissues. Enzyme-linked immunosorbent assay was employed to measure 67 pairs of serum samples from patients and their controls to evaluate its diagnostic value. Immunohistochemistry analysis of 111 formalin-fixed paraffin embedded tissue samples was conducted for explaining its prognostic value. After shRNA transfection, CCK8 and clonal efficiency assays were carried on for verifying the function of IDH1 in vitro. Increased expression at mRNA (P < 0.001) and protein levels (immunohistochemistry: P < 0.001, Western blot analysis: P < 0.001) were observed. Similarly, the IDH1 expression in serum from patients with ESCC was significantly upregulated relative to that from healthy controls (P < 0.001). Kaplan-Meier curve indicated that IDH1 upregulation predicted worse overall survival (OS) and progression-free survival (PFS). Univariate and multivariate analyses identified IDH1 expression as an independent prognostic factor for OS and PFS. Furthermore, OD450 values and colony numbers were decreased in sh-IDH1 groups (all P < 0.05). In conclusion, IDH1 is upregulated in patients with ESCC and can be used as a good potential biomarker for diagnosis and prognosis.

Activity of Glutathione Antioxidant System and NADPH-Generating Enzymes in Rats with Experimental Rheumatoid Arthritis.

Induction of rheumatoid arthritis in rats was accompanied by an increase in diene conjugate content and glutathione reductase and glutathione peroxidase activities in muscles and blood serum. These changes can be related to mobilization of the glutathione reductase/glutathione peroxidase system coupled with intensification of free radical oxidation. In addition, activity of glucose-6-phosphodehydrogenase and NADP-dependent isocitrate dehydrogenase increased, which can be related to increased demand of NADPH for the glutathione reductase/glutathione peroxidase system. The content of reduced glutathione in muscles and blood serum decreased, probably, due to its utilization for ROS neutralization. Glutathione transferase activity decreased in rheumatoid arthritis, which can be related to shortness of reduced glutathione developing during oxidative stress. The observed shifts in parameters of free radical homeostasis in rheumatoid arthritis are probably associated with intensification of free radical oxidation.

Circulating oncometabolite 2-hydroxyglutarate is a potential surrogate biomarker in patients with isocitrate dehydrogenase-mutant intrahepatic cholangiocarcinoma.

PURPOSE: Mutations in the IDH1 and IDH2 (IDH1/2) genes occur in approximately 20% of intrahepatic cholangiocarcinoma and lead to accumulation of 2-hydroxyglutarate (2HG) in the tumor tissue. However, it remains unknown whether IDH1/2 mutations can lead to high levels of 2HG circulating in the blood and whether serum 2HG can be used as a biomarker for IDH1/2 mutational status and tumor burden in intrahepatic cholangiocarcinoma. EXPERIMENTAL DESIGN: We initially measured serum 2HG concentration in blood samples collected from 31 patients with intrahepatic cholangiocarcinoma in a screening cohort. Findings were validated across 38 resected patients with intrahepatic cholangiocarcinoma from a second cohort with tumor volume measures. Circulating levels of 2HG were evaluated relative to IDH1/2 mutational status, tumor burden, and a number of clinical variables. RESULTS: Circulating levels of 2HG in the screening cohort were significantly elevated in patients with IDH1/2-mutant (median, 478 ng/mL) versus IDH1/2-wild-type (median, 118 ng/mL) tumors (P < 0.001). This significance was maintained in the validation cohort (343 ng/mL vs. 55 ng/mL, P < 0.0001) and levels of 2HG directly correlated with tumor burden in IDH1/2-mutant cases (P < 0.05). Serum 2HG levels ≥170 ng/mL could predict the presence of an IDH1/2 mutation with a sensitivity of 83% and a specificity of 90%. No differences were noted between the allelic variants IDH1 or IDH2 in regard to the levels of circulating 2HG. CONCLUSIONS: This study indicates that circulating 2HG may be a surrogate biomarker of IDH1 or IDH2 mutation status in intrahepatic cholangiocarcinoma and that circulating 2HG levels may correlate directly with tumor burden. Clin Cancer Res; 20(7); 1884-90. ©2014 AACR.

Isocitrate dehydrogenase 1 is a novel plasma biomarker for the diagnosis of non-small cell lung cancer.

PURPOSE: Effective biomarkers for the diagnosis of non-small cell lung cancer (NSCLC) are needed. We previously showed that isocitrate dehydrogenase 1 (IDH1) is significantly increased in NSCLC tumors. This study aimed to examine the plasma levels of IDH1 in a large patient population to evaluate its effectiveness in NSCLC diagnosis. EXPERIMENTAL DESIGN: The plasma levels of IDH1, CA125, Cyfra21-1, and CEA were assayed by ELISA. Blood samples were obtained from 1,422 participants (943 patients with NSCLC and 479 healthy controls). The samples were randomly divided into a training set and a test set. Receiver operating characteristic and binary logistic regression analyses were applied to evaluate diagnostic efficacy and establish diagnostic mathematical models. RESULTS: Plasma IDH1 levels were significantly higher in patients with NSCLCs than in healthy controls (P < 0.001). The diagnostic use of IDH1 in lung adenocarcinoma [area under curve (AUC): 0.858 and 0.810; sensitivity: 77.1% and 76.2%; specificity: 82.9% and 76.6%; in the training set and test set, respectively] was significantly greater than that of CA125, Cyfra21-1, or CEA (P < 0.001). The model combining IDH1 with CEA, CA125, and Cyfra21-1 was more effective for lung adenocarcinoma diagnosis than IDH1 alone (sensitivity and specificity in the training set: 75.8%, 89.6%; test set: 86.3%, 70.7%). In addition, the plasma levels of IDH1 could contribute to the diagnostic model of lung squamous cell carcinoma. CONCLUSIONS: IDH1 can be used as a plasma biomarker for the diagnosis of NSCLCs, particularly lung adenocarcinoma, with relatively high sensitivity and specificity.

[Peculiar properties of glutathione system and NADPH-generated enzymes functioning in blood of patients with drug-induced hepatitis under combined treatment with epifamin].

Activity of the glutathione antioxidant system has been studied in patients with drug-induced hepatitis treated with standart base therapy and combined with epifamin therapy. In blood serum of patients before treatment the decrease of reduced glutathione (GSH) level and the increase of glutathione peroxidase (GP) and glutathione reductase (GR) activities versus control were observed. Combined treatment with epifamin changed GSH level to the normal values. Treatment with epifamin was accompanied by pronounced increase in GP and GR activities compared with standard therapy. Activities of the NADPH-generated enzymes, glucose-6-phosphate dehydrogenase and NADP-isocitrate dehydrogenase, which decreased at this pathology, also demonstrated a more significant increase than standart treatment.

Whole-exome sequencing detects somatic mutations of IDH1 in metaphyseal chondromatosis with D-2-hydroxyglutaric aciduria (MC-HGA).

We used exome sequencing of blood DNA in four unrelated patients to identify the genetic basis of metaphyseal chondromatosis with urinary excretion of D-2-hydroxy-glutaric acid (MC-HGA), a rare entity comprising severe chondrodysplasia, organic aciduria, and variable cerebral involvement. No evidence for recessive mutations was found; instead, two patients showed mutations in IDH1 predicting p.R132H and p.R132S as apparent somatic mosaicism. Sanger sequencing confirmed the presence of the mutation in blood DNA in one patient, and in blood and saliva (but not in fibroblast) DNA in the other patient. Mutations at codon 132 of IDH1 change the enzymatic specificity of the cytoplasmic isocitrate dehydrogenase enzyme. They result in increased D-2-hydroxy-glutarate production, α-ketoglutarate depletion, activation of HIF-1α (a key regulator of chondrocyte proliferation at the growth plate), and reduction of N-acetyl-aspartyl-glutamate level in glial cells. Thus, somatic mutations in IDH1 may explain all features of MC-HGA, including sporadic occurrence, metaphyseal disorganization, and chondromatosis, urinary excretion of D-2-hydroxy-glutaric acid, and reduced cerebral myelinization.

Effects of 2,4-dimethoxyphenyl biguanide on glutathione system activity in rat tissues in brain ischemia-reperfusion.

We studied the effects of dimethoxyphenyl biguanide on glutathione peroxidase and glutathione reductase activities, level of reduced glutathione in rat tissues, and activity of some NADPH-regenerating enzymes (glucose-6-phosphate dehydrogenase and NADP-isocitrate dehydrogenase) under conditions of ischemia-reperfusion of the brain. Administration of this biguanide derivative under pathological conditions led to a decrease in the analyzed parameters (elevated under conditions of ischemia-reperfusion) in the serum and brain tissue. The results attest to less pronounced mobilization of the glutathione system (compared to pathological state) due to antioxidant and protective properties of 2,4-dimethoxyphenyl biguanide under conditions of ischemic tissue damage associated with oxidative stress.

Multiple vaccine and pyridostigmine interactions: effects on cognition, muscle function and health outcomes in marmosets.

Following active service during the 1990/1991 Gulf Conflict, a number of UK and US veterans presented with a diverse range of symptoms, collectively known as Gulf Veterans Illnesses (GVI). The administration of vaccines and/or the pretreatment against possible nerve agent poisoning, pyridostigmine bromide (PB), given to armed forces personnel during the Gulf Conflict has been implicated as a possible factor in the aetiology of these illnesses. The possibility that long-term health effects may result from the administration of these vaccines (anthrax, pertussis, plague, yellow fever, polio, typhoid, tetanus, hepatitis B, meningococcal meningitis and cholera) and/or PB, have been investigated using a non-human primate model, the common marmoset. This paper reports the results from three aspects of the study, cognitive behaviour (performance of a touchscreen mediated discrimination task), muscle function (performance of a simple strength test) and general health. There were no marked long-term changes in cognition, muscle function or health that could be attributed to vaccines and/or PB administration. Statistical differences related to treatments were only observed in two aspects of cognition and one of clinical chemistry. These changes were transient in nature and their magnitude were minor and, in consequence, was not regarded as having long-term biological significance.

Metabolism of lymphocytes in mice with Ehrlich ascites carcinoma.

The pattern of metabolic processes in lymphocytes of mice with Ehrlich ascites carcinoma depended on the stage of tumor growth.