Potential Biomarkers For Early Detection Research Articles

N-acety-b-D-glucosaminidase: A potential biomarker for early detection of acute kidney injury in acute chest pain.

Acute kidney injury (AKI) is often underdiagnosed due to several limitations of the renal marker creatinine. Tubular urinary biomarkers may substantially contribute to diagnose AKI early. For early detection of AKI, we evaluated for the first time N-acetyl-β-d-glucosaminidase (NAG), Kidney-injury-molecule-1 (KIM-1) and neutrophil gelatinase-associated lipocalin (NGAL) in acute chest pain. We included 402 chest pain patients aged 18 to 95 years seen in the emergency department. From 311 subjects, blood and urine samples were collected. Thirty-three patients developed an AKI and showed a significant increase in all three tubular markers compared to patients without AKI (each P < .001). According to receiver operating characteristic (ROC) analysis, combining NAG and creatinine showed a significantly increased area under the curve (AUC) compared to creatinine alone (AUC: 0.75 vs 0.87; P < .001). KIM-1, NGAL and cystatin C showed no significant differences in AUC compared to creatinine. In 120 individuals with blood and urine sampling before contrast media exposure, ROC analysis showed a significantly improved diagnostic performance for the combination of both (AUC: 0.83 vs creatinine AUC: 0.66; P = .004). AKI occurrence showed no dependency from CM volume. NAG presented as an independent AKI predictor beside creatinine, age, the diagnosis of myocardial infarction and mean arterial pressure. Regarding the prognostic value for renal replacement therapy, the combination of NAG and creatinine showed a significantly lager AUC than creatinine (AUC: 0.95 vs AUC: 0.85; P < .001). NAG presented as a promising marker of impending AKI and the necessity of renal replacement therapy.

Circulating HSPs Levels and Risk of Human Gastrointestinal Related Cancers: A Systematic Review and Meta-analysis

Heat shock proteins (HSPs) have over-expression in the human malignancies. However, many studies reported inconsistent results. The study aimed to perform a meta-analysis to assess the association between circulating serum HSPs levels and gastrointestinal related cancer risk (GICs). An electronic literature search was conducted for relevant studies in health-related databases up to July 2018. Data related to the standard mean difference (SMD) were collected and analyzed. Summary SMD with 95% confidence intervals (CIs) was estimated using a fixed-effect model. Funnel plot and Egger’s linear regression test were also used for evaluation of publication bias. A total of 11 studies (10 papers) with 584 cases and 576 healthy controls were included. The pooled analysis showed that serum HSPs levels were significantly elevated in subjects with different malignancies than healthy control group (SMD = 0.626; 95% CI 0.507–0.746; P > 0.001). Subgroup analyses by cancer type, HSPs type and sample size also revealed a positive correlation between serum HSPs levels and GICs risk. Elevated serum level of HSPs were strongly associated with colorectal cancer (SMD = 0.696; 95% CI 0.478–0.914; P > 0.001) and pancreatic cancer (SMD = 0.677; 95% CI 0.421–0.933; P > 0.001). This meta-analysis indicated a significant association between high circulating HSPs levels and increased risk of different gastrointestinal malignancies. HSPs may represent a potential biomarker for early detection of cancer patients who may benefit from preventive treatment.

Elevated Expression of Cytosolic Phospholipase A2 Delta Is Associated with Lipid Metabolism Dysregulation during Hepatocellular Carcinoma Progression.

Objective Liver cancer is the third rank amongst the common malignancies, causing maximum death in the patients diagnosed with cancers. Currently available biomarkers are not enough sensitive for early diagnosis of hepatocellular carcinoma (HCC). This makes difficult management of HCC. With the aim of finding new generation of proteomic-based biomarkers, the represented study was designed to characterize the differentially expressed proteins at different stages of HCC initiation and at progression. This could lead to find potential biomarkers for early detection of HCC. Materials and Methods In this experimental study, we report induction of HCC by administrating chemical carcinogens in male Wistar rats. Disease progression was monitored by histological evaluation. Serum proteomic analyses such as 2 dimensional (2D)-electrophoresis, MALDI-TOF-MS/MS and Western blot have been used to analyze and characterize the differentially expressed proteins during HCC development. Results HCC initiation and tumorigenesis were observed at one and four months post carcinogen treatment, respectively. One of the differentially-expressed proteins, namely, cytosolic phospholipase A2delta was significantly up-regulated at very early stage of HCC development. Its expression continued to increase during cancer progression and hepatotumorigenesis stages. Its elevated expression has been confirmed by Western blot analysis. Consistent to this, analyses of the sera in the clinically confirmed liver cancer patients showed elevated expression of this protein, further validating our experimental results. ConclusionThis study suggests that elevation in the expression of cytosolic phospholipase A2delta is associated with progression of HCC.

Braak neurofibrillary tangle staging prediction from in vivo MRI metrics

IntroductionAlzheimer's disease diagnosis requires postmortem visualization of amyloid and tau deposits. As brain atrophy can provide assessment of consequent neurodegeneration, our objective was to predict postmortem neurofibrillary tangles (NFT) from in vivo MRI measurements. MethodsAll participants with neuroimaging and neuropathological data from the Alzheimer's Disease Neuroimaging Initiative, the National Alzheimer's Coordinating Center and the Rush Memory and Aging Project were selected (n = 186). Two hundred and thirty two variables were extracted from last MRI before death using FreeSurfer. Nonparametric correlation analysis and multivariable support vector machine classification were performed to provide a predictive model of Braak NFT staging. ResultsWe demonstrated that 59 of our MRI variables, mostly temporal lobe structures, were significantly associated with Braak NFT stages (P < .005). We obtained a 62.4% correct classification rate for discrimination between transentorhinal, limbic, and isocortical groups. DiscussionStructural neuroimaging may therefore be considered as a potential biomarker for early detection of Alzheimer's disease–associated neurofibrillary degeneration.

Promoter methylation of SEPT9 as a potential biomarker for early detection of cervical cancer and its overexpression predicts radioresistance

BackgroundCervical cancer screening by combined cytology and HPV test has reduced the incidence of cervical cancer, but cytological screening lacks a higher sensitivity while HPV testing possesses a lower specificity. Most patients with invasive cervical cancer are treated with radiotherapy. However, insensitivity to radiotherapy leads to poor efficacy.MethodsIllumina Methylation EPIC 850k Beadchip was used for genomic screening. We detected methylation of SEPT9 and mRNA expression in different cervical tissues by using methylation-specific PCR and qRT-PCR. Then using CCK8, migration assay, and flow cytometry to detect the biological function and irradiation resistance of SEPT9 in vitro and in vivo. Liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) and co-immunoprecipitation (CoIP) were used to find the interacting gene with SEPT9. Immunostaining of CD206 in cervical cancer and polarization of macrophages (M2) were evaluated by immunofluorescence and WB. The Cancer Genome Atlas (TCGA) database was used for screening the potential miRNAs induced by SEPT9.ResultsHyper-methylation of SEPT9 detects cervical cancer and normal tissues, normal+CIN1 and CIN2+CIN3+cancer with high sensitivity and specificity (AUC = 0.854 and 0.797, respectively, P < 0.001). The mRNA and protein expression of SEPT9 was upregulated in cervical cancer tissues when compared to para-carcinoma tissues. SEPT9 promotes proliferation, invasion, migration, and influences the cell cycle of cervical cancer. SEPT9 interacted with HMGB1-RB axis increases irradiation resistance. Furthermore, SEPT9 mediated miR-375 via the tumor-associated macrophages (TAMs) polarization, affecting the resistance to radiotherapy in cervical cancer.ConclusionsThese findings give us the evidence that SEPT9 methylation could be a biomarker for cervical cancer diagnoses. It promotes tumorigenesis and radioresistance of cervical cancer by targeting HMGB1-RB axis and causes polarization of macrophages by mediating miR-375. We suggest SEPT9 could be a potential screening and therapeutic biomarker for cervical cancer.

Inner retinal thinning as a biomarker for cognitive impairment in de novo Parkinson\u2019s disease

We investigated the association between retinal changes measured using optical coherence tomography (OCT) and diverse clinical grading scales in patients with Parkinson’s disease (PD). Seventy-four eyes of 74 patients with de novo PD and 53 eyes of age-matched control subjects were included. The thickness of the peripapillary retinal nerve fiber layer (pRNFL) and macular ganglion cell-inner plexiform layer (mGCIPL) were measured. We analyzed the correlations between the clinical PD grading scales and OCT parameters, and between the OCT parameters and volumetric data in the cerebral cortical and subcortical structures. The area under the receiver operating characteristic curve (AUC) was calculated for diagnosing cognitive impairment in patients with PD. Statistically significant reductions in the thickness of average, temporal, and inferior pRNFL and overall mGCIPL were observed in patients with PD. The Montreal Cognitive Assessment score was significantly associated with mGCIPL thinning. The AUC of the mGCIPL parameters for diagnosing cognitive impairment in patients with PD ranged from 0.651 to 0.760. Moreover, thinning of the mGCIPL was significantly associated with the volumetric parameters of associated brain structures. Our findings highlight the clinical implications of OCT measurements as a potential biomarker for early detection of cognitive impairment in patients with PD.

Tissue inhibitor metalloproteinase-1, Plasminogen activator inhibitor 1, and neutrophil/lymphocyte ratio as potential early biomarkers for diabetic nephropathy

Background: Albuminuria is used to screen early stages of diabetic nephropathy (DN) but it is limited by the fact that structural damage may precede albumin excretion. This necessitates identifying better biomarkers that diagnose or predict diabetic nephropathy. The study aimed to evaluate tissue inhibitor metalloproteinase-1 (TIMP-1), plasminogen activator inhibitor 1 (PAI-1), and neutrophil/lymphocyte ratio (NLR) as potential biomarkers for early detection of diabetic nephropathy and its progression in patients with type 2 diabetes. Materials and Methods: A total of 88 subjects were included in this cross-sectional hospital-based study, healthy individuals (N=10) and diabetic patients (n= 78). Diabetic patients were classified according to an albumin creatinine ratio (ACR) into normoalbuminuria (A1=30), microalbuminuria (A2=14), and macroalbuminuria (A3 =34). TIMP-1, PAI-1 levels, NLR were measured in all subjects. Multivariate discriminant analysis (MDA) was used to develop a novel index. The diagnostic value of TIMP-1, PAI-1, and NLR was assessed by the area under the receiver operating characteristic (ROC). Results: The mean ± SD of NLR, PAI-1 (ng/ml) and TIMP-1 (ng/ml) in healthy were (1.9±0.30; 6.8 ±2.4 and 76.8±15.7) and in A1 were (3.0 ± 2.5;7.7±1.8 and 91.4±19.8) and in A2 were ( 2.2±1.5; 7.6±1.9 and 104.5±20.9) and in A3 were (5.2±3.9; 8.7±1.2 and 120.6±18.2). The differences between the mean of NLR, PA1-1 and, TIMP in A2 and that of A3 were significant (p <0.007, p =0.03 and, p < 0.011; respectively). TIMP-1 was the most efficient marker with an AUC of 0.72 for discriminant diabetics with A1 from A2; 0.88 for A1 vs A3 and 0.82 for A2 vs A3. A novel index was developed for differentiated between stages of DN based on three blood markers (TIMP-1, PAI-1, and NLR) named TPN. The mean ± SD of TPN index in healthy was (1.1±0.4) and in A1 was (1.2 ±0.3) and in A2 was (1.3 ±0.2) and in A3 were (1.8±0.3) with a highly significant difference between A2 and A3. The AUC of the TPN index was 0.61, 0.88, and 0.88 for discriminant diabetics with A1 from A2, A1 vs A3, and A2 vs A3. Conclusions: A novel index named TPN based on three blood markers (TIMP-1, PAI-1, and NLR) may be potentially useful for early detection and to discriminate macro-albuminuria from micro-albuminuria stages.

Circulating microRNAs as Potential Diagnostic and Prognostic Biomarkers in Hepatocellular Carcinoma

Hepatocellular carcinoma (HCC) is the fifth most common cancer with high mortality, due to late diagnosis and limited treatment options. Blood miRNAs, which circulate in a highly stable, cell-free form, show promise as novel potential biomarkers for early detection of HCC. Whole miRNome profiling was performed to identify deregulated miRNAs between HCC and normal healthy (NH) volunteers. These deregulated miRNAs were validated in an independent cohort of HCC, NH and chronic Hepatitis B (CHB) volunteers and finally in a 3rd cohort comprising NH, CHB, cirrhotic and HCC volunteers to evaluate miRNA changes during disease progression. The associations between circulating miRNAs and liver-damage markers, clinicopathological characteristics and survival outcomes were analysed to identify prognostic markers. Twelve miRNAs are differentially expressed between HCC and NH individuals in all three cohorts. Five upregulated miRNAs (miR-122-5p, miR-125b-5p, miR-885-5p, miR-100-5p and miR-148a-3p) in CHB, cirrhosis and HCC patients are potential biomarkers for CHB infection, while miR-34a-5p can be a biomarker for cirrhosis. Notably, four miRNAs (miR-1972, miR-193a-5p, miR-214-3p and miR-365a-3p) can distinguish HCC from other non-HCC individuals. Six miRNAs are potential prognostic markers for overall survival.

Abstract 3329: Lung cancer screening initiative and identification of novel blood biomarkers for early detection of lung cancer

Abstract In lung cancer, 81% of patients detected are late-stage, since early-stage patients are generally asymptomatic. Winnebago County has a 14% higher mortality rate for lung cancer when compared to the national rate; hence, to improve early detection of lung cancer in the Winnebago County, we implemented low-dose computed tomography (LDCT) screening. Currently, LDCT is used to detect lung cancer at early-stages, and the CDC has proposed new guidelines for early screening of lung cancer in individuals between 55-77 years of age with a 30 pack/year smoking history. Using circulating DNA from lung cancer patients and examining changes in methylation patterns in genes that are expressed in lung cancer patients can be used for early detection. Epidermal growth factor receptor (EGFR) is shown to be expressed in lung cancer patients’ serum in all stages; hence, it can be used as a potential biomarker for early detection. To improve early detection of lung cancer in the Winnebago County, we educated physicians/smokers about LDCT screenings to promote early detection of lung cancer. We evaluated the number of LDCTs performed and recorded the number of lung cancer cases detected in the Winnebago County between June 2015 to October 2017. To explore EGFR as a promising biomarker, plasma was separated from whole blood and collected. Circulating DNA was extracted with the MagMax cell-free circulating kit and bisulfite converted. Identification of percent methylation patterns in the EGFR promoter region in CpG islands was obtained after next generation sequencing (NGS) followed by bioinformatics analysis. In our study, 1,116 LDCT screenings occurred in the Winnebago County. Lung cancer was diagnosed in 19 patients, out of which 11 patients (57.8%) were early-stage patients. We detected 45.4% of patients screened that were present with nodules compared to 24.2% of patients in the National Lung Cancer Screening Trial. We observed that 1.70% of patients referred for LDCT screening were diagnosed with lung cancer, compared to 0.87% of patients in the National Lung Cancer Screening Trial. This may be due to the higher incidence of smoking in Winnebago County (18%) compared to the nation (15.5%). Our screening study had 96.3% false positives, which is comparable (96.4%) to the results obtained in the National Lung Cancer Screening Trial. Using NGS, methylation in the promoter region of EGFR was studied using degenerate EGFR primers. In the promoter region of EGFR, our analysis showed that there are regions of hypermethylation (68-86%) in three CpG islands in sequences at the beginning of the promoter (8-241 bases). However, there was hypomethylation (6-21%) at four CpG island further downstream (375-689 bases from the beginning of the promoter). Currently we are studying the methylation patterns in normal subjects to determine whether these regions which could be used as predictive biomarkers for early detection. Citation Format: Adijan Kuckovic, Joseph Berei, Shylendra Sreenivasappa, Joseph Ross, Sandra Martell, Connie Vitali, William Schulz, Neelu Puri. Lung cancer screening initiative and identification of novel blood biomarkers for early detection of lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3329.

Abstract 415: Ultrasensitive analysis of circulating exosomes for liquid biopsy-based cancer diagnosis using 3D nano-engineered microfluidic chips

Abstract Liquid biopsy is extremely appealing in early diagnosis, prognosis, and precision treatment of cancer, as tissue biopsy is highly invasive, costly, and often infeasible to repeat. Extracellular vesicles (EVs), including exosomes, are emerging as a new paradigm of liquid biopsy for cancer diagnosis and monitoring response to therapy. Current affinity isolation and analysis methods rely on surface capture of EVs which is governed by the fundamental limits in mass transfer, surface binding reaction, and the boundary effects due to the hydrodynamic resistance at the liquid-solid interface. A major conceptual constraint in existing techniques is that they only partially addressed these fundamental factors. Here we report an innovative strategy for 3D nano-engineering of microfluidic chip to address the major limits in one device. This lithography-free engineering strategy exploits microfabricated channels to guide nanoparticle self-assembly, enabling simple, large-scale integration of self-assembled microelements with unique 3D nanostructures for biosensing. Using this method, we devised a nanoporous herringbone (nano-HB) chip that presents distinct advantages to address the aforementioned limits in one device, as it 1) effectively promotes microscale mass transfer; 2) increases the surface area and probe density to enhance binding efficiency and speed; and 3) reduces near-surface hydrodynamic resistance to promote particle-surface interactions for binding. Using the nano-HB chip, we demonstrated highly efficient (&amp;gt;80%) isolation of EVs purified from COLO-1, MCF-7, and SKOV3 cell lines spiked in healthy plasma. The nano-HB chip was further validated for elution and downstream analysis of captured EVs, i.e., targeted mRNA profiling by droplet digital PCR (ddPCR) and protein profiling by microplate ELISA. Assessed with spiked-in EV standards, the nano-HB chip was seen to afford an extremely low limit of detection of 10 μL-1 (200 vesicles per assay) and excellent compatibility with complex blood plasma. Such analytical capabilities enabled ultrasensitive and quantitative detection of low-level folate receptor alpha (FRα) positive exosomes in ovarian cancer patient plasma that are otherwise undetectable to standard assays. We demonstrated sensitive and specific detection of exosome subpopulations expressing CD24, EpCAM, and FRα proteins for non-invasive diagnosis of ovarian cancer using only 2 μL plasma. Comparative evaluation showed that our nano-HB chip greatly improves diagnostic power over the standard microplate ELISA and the combined nano-HB capture and ddPCR analysis of mRNAs. Our findings suggest exosomal FRα as a potential biomarker for early detection and monitoring of progression of ovarian cancer. Our technology would offer enabling exosome isolation and sensing abilities to facilitate liquid biopsy-based cancer diagnosis. Citation Format: Peng Zhang, Xin Zhou, Yong Zeng. Ultrasensitive analysis of circulating exosomes for liquid biopsy-based cancer diagnosis using 3D nano-engineered microfluidic chips [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 415.

Gene expression analysis of MMPs in women with preeclampsia using cell-free fetal RNA in maternal plasma

Nucleic acids released from the placenta into the mother's blood circulation system provide a valuable source of potential biomarkers for early detection of pregnancy complications such as preeclampsia (PE). PE affects nearly 5-10% of pregnancies worldwide and is a major contributor to the maternal and neonatal mortality and morbidity. It is known that altered placental expression of matrix metalloproteinases (MMPs) may cause shallow cytotrophoblastic invasion and ultimately lead to preeclampsia. The present study aimed to evaluate pattern of placental/fetal expression of the MMP family (MMP-2, MMP-9, MMP-14, MMP-15 and MMP-26) in preeclamptic women and compare it to normal pregnancies, using cell free fetal RNA (cff-RNA). Blood samples were obtained from 20 pregnant women diagnosed with severe PE (28-32 weeks) and 40 control healthy pregnant women in two groups of either matched gestational age (N = 20) or 14 and 28 weeks pregnancies (each 10). cff-RNA was extracted from plasma, followed by reverse transcription of cff-RNA. Expression of MMP genes was measured using quantitative reverse transcription PCR (qRT-PCR). The expression levels of MMP-2, MMP-9 and MMP-15 were significantly increased, while MMP-14 expression level was significantly reduced and the expression of MMP-26 showed a relative increase in PE pregnancies compared to the control group. Additionally, increased level of MMPs expression was observed by comparing 14 and 28 weeks gestation age in normal pregnancy. Using cff-RNA, circulatory expression level of MMP-2, MMP-9, MMP-14 and MMP-15 were significantly altered in preeclampsia compared to normal pregnancies.

TGF-β1 and VCAM-1 Serum Concentrations as Diagnostic Biomarkers of Diabetic Kidney Disease Progression

Abstract Background and aims: Transforming growth factor-beta 1 (TGF-β1) and vascular adhesion molecule 1 (VCAM-1) have been proposed as promising biomarkers for multiple diseases. TGF-β1 and VCAM-1 are reported to be associated with diabetic kidney disease (DKD) and end stage renal disease in patients with type 1 and type 2 diabetes mellitus (T1DM and T2DM). Material and methods: The aim of this study was to investigate the expression of circulating TGF-β1 and VCAM-1 and to assess their potential as a blood-based biomarker for DKD in T1DM and T2DM patients. Results:. The study included 124 participants: 66 patients with T1DM, 58 with T2DM and 20 healthy controls. The diabetic patients were classified according to the estimated glomerular filtration rate (eGFR). First group - eGFR ≥90ml/min/1.73 m2 (n=39), second group eGFR 89-60 ml/min/1.73m2 (n=45), and third group eGFR 59-45 ml/min/1.73m2 (n=40). Enzyme-linked immunosorbent assay for the quantitative detection of was used to evaluate blood TGF-β1 and VCAM-1 expression. It was found that there were higher TGF-β1 and VCAM-1 in all diabetic patients compared with healthy controls (P&lt;0.05). TGF- β1 and VCAM-1 were higher in group with eGFR ≥90ml/min/1.73 m2 and gradually increased in the groups with eGFR89-60 ml/min/1.73m2 and eGFR 59-45 ml/min/1.73m2. TGF- β1 and VCAM-1 were less in T1DM, than T2DM in all study groups. Regression analysis revealed reverse associations between TGF- β1, VCAM-1 and eGFR (P&lt;0.05). TGF- β1 and VCAM-1 correlated positively with albuminuria and negatively with renal function. Conclusion: In discriminating overall patients from healthy subjects, ROC analysis revealed areas under the curve (AUCs) of 1,0 for TGF- β1 for T1DM and T2DM, VCAM-1 0,866 for T1DM, 0,923 for T2DM (P&lt;0.001). The results suggested that blood-based TGF- β1 and VCAM-1 may serve as potential biomarkers for early detection of DKD.

Circulating miRNA-21 and miRNA-23a Expression Signature as Potential Biomarkers for Early Detection of Non-Small-Cell Lung Cancer.

Lung Cancer (LC) is a major cancer killer worldwide, and 5-yr survival is extremely poor (≤15%), accentuating the need for more effective diagnostic and therapeutic strategies. Studies have shown cell-free microRNAs (miRNAs) circulating in the serum and plasma with specific expression in cancer, indicating the potential of using miRNAs as biomarkers for cancer diagnosis and therapy. This study aimed to identify differentially-expressed two miRNAs in the plasma of Non-Small Cell Lung Cancer (NSCLC) patients that might be a clinically useful tool for lung cancer early detection. miRNA-21 is one of the most abundant oncomirs. miRNA-23a functions as an oncogene in several human cancers, however, its clinical value has not been investigated in NSCLC. A case-control study was conducted in Assiut University Hospital, Egypt, from 2017 to 2018. Plasma samples were obtained from 45 NSCLC patients. The expression level of miR-21 and miRNA-23a was detected by qRT-PCR and compared to 40 healthy control subjects. The relation between both miRNAs and clinicopathological parameters was evaluated. The expression level of miR-21 and miRNA-23a was significantly up-regulated (36.9 ± 18.7 vs. 1.12 ± 0.84 and 24.7 ± 19.09 vs. 1.16 ± 0.45) in NSCLC compared to matched controls (P<0.0001each). There was a significant difference in the level of plasma miRNA-21 and miRNA- 23a expression between the different grades of the disease (P = 0.032 and P = 0.001, respectively). The plasma miRNA-21 and miRNA-23a levels in the lung cancer patients with distant metastasis (n = 20) were significantly higher than those in the patients without metastasis (n = 25) (P<0.0001 each), the expression of miR-21 and miRNA-23a was significantly associated with tumor size (P = 0.001, P = 0.0001, respectively), but not significantly related to lymph node metastasis (P = 0.687 and 0.696, respectively). A positive correlation was observed between miRNA-21 and miRNA-23a (r = 0.784, P<0.01), There was no significant difference in the plasma miRNA-21 and miRNA-23a levels in the lung cancer patients with different histopathological types. miR-21 and miR-23a might play an oncogenic role in LC and is a poor prognostic factor. Switching off miRNA-21 and miRNA-23a may improve the treatment of LC. Our results must be verified by large-scale prospective studies with standardized methodology.

New Platform for the Direct Profiling of microRNAs in Biofluids.

Circulating microRNAs have been identified as potential biomarkers for early detection, prognosis, and prediction of several diseases. Their use in clinical diagnostics has been limited by the lack of suitable detection techniques. Most of the current technologies suffer from requiring complex protocols, not yet able to deliver robust and cost-effective assays in the field of clinical diagnostics. In this work, we report the development of a breakthrough platform for profiling circulating microRNAs. The platform comprises a novel silicon photomultiplier-based reader in conjunction with a chemical-based method for nucleic acid detection. Accurate microRNAs profiling without extraction, pre-amplification, or pre-labeling of the target is now achievable. We designed and synthesized a set of reagents that combined the chemical-based method with a chemiluminescent reaction. The signals generated were read outusing a novel, compact silicon photomultiplier-based reader. The platform sensitivity was determined by measuring known concentrations of hsa-miR-21-5p spike-ins. The limit of detection was calculated as 4.7 pmol/L. The platform was also successfully used to directly detect hsa-miR-21-5p in eight non-small cell lung cancer plasma samples. Levels of plasma hsa-miR-21-5p expression were also measured via TaqMan RT-qPCR. The successful integration of theunique chemical-based method for nucleic acid detection with the novel silicon photomultiplier-based reader created an innovative product (ODG platform) with diagnostic utility, for the direct qualitative and quantitative analysis of microRNA biomarkers in biological fluids.

Promoter methylation of ADAMTS1 and BNC1 as potential biomarkers for early detection of pancreatic cancer in blood

BackgroundDespite improvements in cancer management, most pancreatic cancers are still diagnosed at an advanced stage. We have recently identified promoter DNA methylation of the genes ADAMTS1 and BNC1 as potential blood biomarkers of pancreas cancer. In this study, we validate this biomarker panel in peripheral cell-free tumor DNA of patients with pancreatic cancer.ResultsSensitivity and specificity for each gene are as follows: ADAMTS1 87.2% and 95.8% (AUC = 0.91; 95% CI 0.71–0.86) and BNC1 64.1% and 93.7% (AUC = 0.79; 95% CI 0.63–0.78). When using methylation of either gene as a combination panel, sensitivity increases to 97.3% and specificity to 91.6% (AUC = 0.95; 95% CI 0.77–0.90). Adding pre-operative CA 19-9 values to the combined two-gene methylation panel did not improve sensitivity. Methylation of ADAMTS1 was found to be positive in 87.5% (7/8) of stage I, 77.8% (7/9) of stage IIA, and 90% (18/20) of stage IIB disease. Similarly, BNC1 was positive in 62.5% (5/8) of stage I patients, 55.6% (5/9) of stage IIA, and 65% (13/20) of patients with stage IIB disease. The two-gene panel (ADAMTS1 and/or BNC1) was positive in 100% (8/8) of stage I, 88.9% (8/9) of stage IIA, and 100% (20/20) of stage IIB disease. The sensitivity and specificity of the two-gene panel for localized pancreatic cancer (stages I and II), where the cancer is eligible for surgical resection with curative potential, was 94.8% and 91.6% respectively. Additionally, the two-gene panel exhibited an AUC of 0.95 (95% CI 0.90–0.98) compared to 57.1% for CA 19-9 alone.ConclusionThe methylation status of ADAMTS1 and BNC1 in cfDNA shows promise for detecting pancreatic cancer during the early stages when curative resection of the tumor is still possible. This minimally invasive blood-based biomarker panel could be used as a promising tool for diagnosis and screening in a select subset of high-risk populations.

Whole-Blood DNA Methylation Markers in Early Detection of Breast Cancer: A Systematic Literature Review.

Whole-blood DNA methylation markers have been suggested as potential biomarkers for early detection of breast cancer. We conducted a systematic review of the literature on whole-blood DNA methylation markers for breast cancer detection. PubMed and ISI Web of Knowledge were searched up to May 29, 2018. Overall, 33 studies evaluating 355 markers were included. The diagnostic value of most individual markers was relatively modest, with only six markers showing sensitivity >40% at specificity >75% [only 2 (HYAL2 and S100P) were independently validated]. Although relatively strong associations (OR ≤0.5 or OR ≥2) with breast cancer were reported for 14 markers, most of them were not independently validated. Two prospective studies performed epigenome-wide association analysis and identified 276 CpG sites related to breast cancer risk, but no overlap was observed between CpGs reported from these two studies. Five studies incorporated individual markers as panels, but only two of them used a test-validation approach. In conclusion, so far detected methylation markers are insufficient for breast cancer early detection, but markers or marker-combinations may be useful for breast cancer risk stratification. Utilizing high-throughput methods of methylation quantification, future studies should focus on further mining informative methylation markers and derivation of enhanced multimaker panels with thorough external validation ideally in prospective settings.

Ultrasensitive detection of circulating exosomes with a 3D-nanopatterned microfluidic chip.

The performance of current microfluidic methods for exosome detection is constrained by boundary conditions and by fundamental limits to microscale mass transfer and to interfacial exosome binding. Here, we show that a microfluidic chip designed with self-assembled 3D herringbone nanopatterns can detect low levels of tumour-associated exosomes in plasma (10 exosomes μL−1, or approximately 200 vesicles per 20-μL spiked sample) that would otherwise be undetectable by standard microfluidic systems for biosensing. The nanopatterns promote microscale mass transfer, increase surface area and probe density to enhance the efficiency and speed of exosome binding, and permit drainage of the boundary fluid to reduce near-surface hydrodynamic resistance, thus promoting particle–surface interactions for exosome binding. We used the device for the detection, in 2-μL plasma samples from 20 ovarian cancer patients and from 10 age-matched controls, of exosome subpopulations expressing CD24, EpCAM, and FRalpha proteins, and suggest exosomal FRalpha as a potential biomarker for the early detection and progression monitoring of ovarian cancer. The nanolithography-free nanopatterned device should facilitate the use of liquid biopsies for cancer diagnosis.

Current state of knowledge on the health effects of engineered nanomaterials in workers: a systematic review of human studies and epidemiological investigations.

Objectives The widespread application of nano-enabled products and the increasing likelihood for workplace exposures make understanding engineered nanomaterial (ENM) effects in exposed workers a public and occupational health priority. The aim of this study was to report on the current state of knowledge on possible adverse effects induced by ENM in humans to determine the toxicological profile of each type of ENM and potential biomarkers for early detection of such effects in workers. Methods A systematic review of human studies and epidemiological investigations of exposed workers relative to the possible adverse effects for the most widely used ENM was performed through searches of major scientific databases including Web of Science, Scopus, and PubMed. Results Twenty-seven studies were identified. Most of the epidemiological investigations were cross-sectional. The review found limited evidence of adverse effects in workers exposed to the most commonly used ENM. However, some biological alterations are suggestive for possible adverse impacts. The primary targets of some ENM exposures were the respiratory and cardiovascular systems. Changes in biomarker levels compared with controls were also observed; however, limited exposure data and the relatively short period since the first exposure may have influenced the incidence of adverse effects found in epidemiological studies. Conclusions There is a need for longitudinal epidemiologic investigations with clear exposure characterizations for various ENM to discover potential adverse health effects and identify possible indicators of early biological alterations. In this state of uncertainty, precautionary controls for each ENM are warranted while further study of potential health effects continues.

Evaluation of urinary vitamin D-binding protein in type 1 diabetic children

Objective The aim of the study was to measure the levels of vitamin D-binding protein (VDBP) and microalbumin in the urine of diabetic children (type 1) and to analyze the correlation of VDBP with other parameters. Background Elevated urinary vitamin D-binding protein (UVDBP) level in patients with diabetes indicates that renal tubular damage may be involved in early stages of the development of diabetic nephropathy (DN). Patients and methods A total of 20 type 1 diabetic patients and 20 healthy controls were subjected to full history taken, thorough clinical examination, and laboratory investigation, which included complete blood count, blood glucose profile, glycosylated hemoglobin, parameters of kidney function, liver function, measurement of microalbuminurea, and VDBP in urine by enzyme-linked immunosorbent assay. Results UVDBP level showed significant increases in type 1 diabetic patients with microalbuminuria and macroalbuminuria (mean: 562.990 ± 194.771) when compared with control group (mean: 172. 480 ± 41.856; P = 0.001). There was a positive correlation between UVDBP as a marker of DN and fasting blood glucose, 2-h postprandial blood glucose (2-h PP), glycosylated hemoglobin, duration of the disease, and albumin/creatinine ratio, but no significant correlation between UVDBP as a marker of DN and other laboratory data. Conclusion Our finding indicates that UVDBP level is a potential biomarker for early detection of DN.

Circulating visfatin levels and cancers risk: A systematic review and meta-analysis.

Visfatin levels have been reported to be abnormal in many types of cancers. However, epidemiological studies yielded inconsistent results. Therefore, a meta-analysis was performed to assess the association between circulating visfatin levels and cancer risk. A systematic search was conducted for relevant studies in health-related electronic databases up to March 2018. Data related to standard mean difference (SMD) and overall odds ratio (ORS) were collected and analyzed. Summary SMD and pooled OR with 95% CIs were calculated using a random-effect model. Funnel plot and Egger's linear regression test were conducted to examine the risk of publication bias. A total of 27 studies with 2,693 cases and 3,040 healthy controls were included in meta-analysis for pooling SMD analysis. The results of the meta-analysis showed a significant higher visfatin levels in patients with various cancers than in controls, with a pooled SMD of 0.88, 95% CI = 0.56-1.20, p = 0.000. In subgroup, metaregression, Galbraith plot, and sensitivity analysis showed no substantial difference among all the analyzed factors. Data from 14 studies were also used for pooling ORs analysis. Metaresults revealed that high visfatin levels were associated with cancer risk (OR = 1.24, 95% CI: 1.14-1.34, p = 0.000). No evidence of publication bias was observed for pooling ORs and SMD analysis. This meta-analysis indicated a significant association between high circulating visfatin levels and increased risk of various cancers. Visfatin may represent a potential biomarker for early detection of cancers who may benefit from preventive treatment.Note.