Non-invasive Early Diagnosis Research Articles

MOLECULAR MECHANISMS OF FORMATION OF CHRONIC PELVIC PAIN SYNDROME IN PATIENTS WITH EXTERNAL GENITAL ENDOMETRIOSIS: MARKERS AND TARGETS FOR DIAGNOSIS AND THERAPY (REVIEW)

This article systematizes the results of studies on the pathogenetic mechanisms of CPPS formation in patients with EGE. Pain is one of the severe clinical manifestations and complications of endometriosis. To ease the pain in patients with EGE is complex but urgent task. Long diagnostic search (mean period 5−8 years) leads to the formation of a persistent Autonomous pain syndrome associated with relapses of pain even after a com-prehensive treatment of patients (surgical and/or long-term hormonal). Patients with endometriosis suffer from mixed pain including neuropathic and nociceptive components which is proved by the example of expression in ectopic foci of specific biomarkers. Therefore, the question of early non-invasive diagnosis of endometriosis is very vital and can be solved by the development of a diagnostic panel consisting of biomarkers expressed by ectopic foci at the initial stage of the disease.

Targeting Detection and Inhibition of Methicillin-Resistant Staphylococcus aureus Pneumonia with a Theranostic Mesoporous Silica-Based Platform.

In this study, non-invasive early stage diagnosis of methicillin-resistant Staphylococcus aureus-related pneumonia and related anti-inflammatory profiles with theranostic platforms were reported. Mesoporous silica NPs were first synthesized and chemically modified to bear an amino group and further covalently bind to amino-2,4,6-triiodoisophthalic acid and anti-methicillin-resistant S. aureus antibodies. The drug-loaded NPs were characterized by transmission electron microscopy, dynamic light scattering analysis and X-ray photoelectron spectra. MTT and broth dilution assays were used to investigate the biocompatibility and antibacterial potency of the platform. In vivo computed tomography analysis was performed in rats infected with different kinds of pneumonia, and anti-inflammatory studies such as bronchoalveolar lavage and pathological assays were used to test the capability of our nanomedicine as a theranostic platform. In addition, the long-term safety of the hybrid NPs was investigated via hematological and biochemical assays.

Abstract 1580: Separation of circulating biomarkers by nanoDLD lab-on-a-chip technology

Abstract Efficient isolation of circulating biomarkers is key for enabling liquid biopsies for cancer diagnosis and prognosis. Circulating biomarkers such as exosomes and cell-free DNA hold promise for non-invasive early cancer diagnosis, determining cancer stage and prognosis, as well as monitoring treatment progression and the development of drug resistance. However, isolation of these biomarkers has proven challenging in the clinic. Exosomes range in size from approximately 30 to 150 nm and the current gold standard isolation method includes large-scale ultracentrifugation, which is time-consuming and lacks reproducibility. Circulating cell-free DNA is found in a variety of sizes and states and while clinical applications for prenatal testing have emerged, applications in oncology remain elusive in part due to high background and challenges in isolating relevant molecules. We have developed a lab-on-a-chip technology based on deterministic lateral displacement at the nanoscale (nanoDLD) which separates and concentrates particles as small as 20 nm from smaller particles in continuous flow. Analysis of nanoDLD isolation of exosomes and DNA show improved resolution, recovery, and concentration compared to standard techniques. Current efforts to scale nanoDLD technology to clinically relevant volumes will provide a reproducible and automated tool for isolation, study, and clinical use of these important non-invasive biomarkers. Citation Format: Stacey M. Gifford, Benjamin H. Wunsch, Joshua T. Smith, Navneet Dogra, Sungcheol Kim, Gustavo Stolovitzky. Separation of circulating biomarkers by nanoDLD lab-on-a-chip technology [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 1580.

Fe-Au Nanoparticle-Coupling for Ultrasensitive Detections of Circulating Tumor DNA.

Effectiveness of cancer therapy relies heavily on the efficient early diagnosis. Circulating tumor DNA (ctDNA) detection is one of the most clinically meaningful liquid biopsy approaches for the noninvasive cancer early diagnosis, which, unfortunately, cannot be applied as a routine diagnostic tool till a number of obstacles, for example, unsatisfactory specificity and sensitivity, and extremely high costs, are overcome. Here, the first paradigm of nanomaterial's application in the extremely specific, ultrasensitive, and yet economical ctDNA detections is reported based on a facile nanoparticle-coupling strategy without amplification, with which polymerase chain reaction (PCR)-introduced bias and other shortcomings are successfully circumvented. Aiming at seven Kirsten rat sarcoma-2 virus (KRAS) point mutations, the present strategy exhibits high specificity and an ultrahigh sensitivity of detecting as low as 0.1 pg mL-1 of KRAS point mutation without prior PCR amplification. Discriminating KRAS gene mutations in lung adenocarcinoma patients at an extremely low detection limit equivalent to 0.12% mutation relative to wild-type gene is successful. It is envisioned that this nanoparticle-coupling approach could be routinely applied clinically for ultra-early diagnosis and monitoring of diverse malignant tumors, thus facilitating the fight against cancer.

Analysis of Different Methods for the Diagnosis of Subclinical and Clinical Stages Polyneuropathy in Patients with Type 2 Diabetes Mellitus

Many patients with T2DM have a late complication at the time of diagnosis,most commonly diabetic peripheral neuropathy (DPN).We explored whether clinical indiced of polyneuropathy correlate with large and small nerve fiber pathology. The goal of this study is to analyzed various diagnostic techniques for the detection of lesions of nerve fibers in patients with T2DM and subclinical and clinical stages of DNP by electrophysiology study and nerve conduction velocity (NCV),skin biopsy and corneal confocal microscopy (CCM) with use of the author's algorithm for calculating the tortuosity and orientation of the nerve fibers of the cornea. Materials and methods: Electrophysiological study of large nerve fibers and microscopic examination of small nerve fibers in patients with T2DM with subclinical (n=50) and clinical stages of DPN (n=50) were performed. Results: The changes in the S-response and in NCV were received the most frequently in patients with subclinical and clinical stages of DPN in the study of the sensory gastrocnemius nerve.At stimulation of the motor nerves,the changes were related to the M-response and NCV of the peroneal and tibial nerves.At skin biopsy,we saw that the intraepidermal nerve fibers density (IENFD) in patients with T2DM was significantly reduced in subclinical and clinical stages of DPN.In the subclinical form of DPN,some values of CCM significant correlate with IENFD and with NCV along the gastrocnemius nerve in the entire studied population,but this correlation was not strong.At symptomatic DPN,a strong correlation was found between IENFD and parameters of NCV (p<0.05),as well as with data of electroneyromyograpry (p<0.05). Conclusion: The results of the study showed that the changes in large fiber function accompany the loss of small nerve fibers and occurs in patients with T2DM already in the preclinical stage of DPN. CCM can be a powerful tool for non-invasive early diagnosis of neuropathy in subclinical stage DPN. Disclosure A. Fokina: None. A. Zilov: None. I. Strokov: None. Z. Surnina: None.

Highly accurate model for prediction of lung nodule malignancy with CT scans

Computed tomography (CT) examinations are commonly used to predict lung nodule malignancy in patients, which are shown to improve noninvasive early diagnosis of lung cancer. It remains challenging for computational approaches to achieve performance comparable to experienced radiologists. Here we present NoduleX, a systematic approach to predict lung nodule malignancy from CT data, based on deep learning convolutional neural networks (CNN). For training and validation, we analyze >1000 lung nodules in images from the LIDC/IDRI cohort. All nodules were identified and classified by four experienced thoracic radiologists who participated in the LIDC project. NoduleX achieves high accuracy for nodule malignancy classification, with an AUC of ~0.99. This is commensurate with the analysis of the dataset by experienced radiologists. Our approach, NoduleX, provides an effective framework for highly accurate nodule malignancy prediction with the model trained on a large patient population. Our results are replicable with software available at http://bioinformatics.astate.edu/NoduleX.

Microbial markers in colorectal cancer detection and/or prognosis.

Colorectal cancer (CRC) is the second leading cause of cancer worldwide. CRC is still associated with a poor prognosis among patients with advanced disease. On the contrary, due to its slow progression from detectable precancerous lesions, the prognosis for patients with early stages of CRC is encouraging. While most robust methods are invasive and costly, actual patient-friendly screening methods for CRC suffer of lack of sensitivity and specificity. Therefore, the development of sensitive, non-invasive and cost-effective methods for CRC detection and prognosis are necessary for increasing the chances of a cure. Beyond its beneficial functions for the host, increasing evidence suggests that the intestinal microbiota is a key factor associated with carcinogenesis. Many clinical studies have reported a disruption in the gut microbiota balance and an alteration in the faecal metabolome of CRC patients, suggesting the potential use of a microbial-based test as a non-invasive diagnostic and/or prognostic tool for CRC screening. This review aims to discuss the microbial signatures associated with CRC known to date, including dysbiosis and faecal metabolome alterations, and the potential use of microbial variation markers for non-invasive early diagnosis and/or prognostic assessment of CRC and advanced adenomas. We will finally discuss the possible use of these markers as predicators for treatment response and their limitations.

Novel non-invasive early detection of lung cancer using liquid immunobiopsy metabolic activity profiles.

Lung cancer is the leading cause of cancer death worldwide. Survival is largely dependent on the stage of diagnosis: the localized disease has a 5-year survival greater than 55%, whereas, for spread tumors, this rate is only 4%. Therefore, the early detection of lung cancer is key for improving prognosis. In this study, we present an innovative, non-invasive, cancer detection approach based on measurements of the metabolic activity profiles of immune system cells. For each Liquid ImmunoBiopsy test, a 384 multi-well plate is loaded with freshly separated PBMCs, and each well contains 1 of the 16 selected stimulants in several increasing concentrations. The extracellular acidity is measured in both air-open and hermetically-sealed states, using a commercial fluorescence plate reader, for approximately 1.5h. Both states enable the measurement of real-time accumulation of 'soluble' versus 'volatile' metabolic products, thereby differentiating between oxidative phosphorylation and aerobic glycolysis. The metabolic activity profiles are analyzed for cancer diagnosis by machine-learning tools. We present a diagnostic accuracy study, using a multivariable prediction model to differentiate between lung cancer and control blood samples. The model was developed and tested using a cohort of 200 subjects (100 lung cancer and 100 controlsubjects), yielding 91% sensitivity and 80% specificity in a 20-fold cross-validation. Our results clearly indicate that the proposed clinical model is suitable for non-invasive early lung cancer diagnosis, and is indifferent to lung cancer stage and histological type.

Simultaneous Quantification of Multiple Cancer Biomarkers in Blood Samples through DNA-Assisted Nanopore Sensing.

Protein biomarkers in blood have been widely used in the early diagnosis of disease. However, simultaneous detection of many biomarkers in a single sample remains challenging. Herein, we show that the combination of a sandwich assay and DNA-assisted nanopore sensing could unambiguously identify and quantify several antigens in a mixture. We use five barcode DNAs to label different gold nanoparticles that can selectively bind specific antigens. After the completion of the sandwich assay, barcode DNAs are released and subject to nanopore translocation tests. The distinct current signatures generated by each barcode DNA allow simultaneous quantification of biomarkers at picomolar level in clinical samples. This approach would be very useful for accurate and multiplexed quantification of cancer-associated biomarkers within a very small sample volume, which is critical for non-invasive early diagnosis of cancer.

Risk factors for recurrence of external genital endometriosis after surgical treatment: prospective cohortant study

Objectives of the study: improve early non-invasive diagnosis of external genital endometriosis (EGE) using the mass spectrometry method, and determine the risk factors for recurrence of EGE after surgical treatment. Material and methods: a prospective cohort study included 100 patients with EGE who underwent surgical treatment during the 2014 to 2016 period. The EGE diagnosis was made on the basis of diagnostic and therapeutic laparoscopic findings and finally confirmed at histological exam. All patients underwent blood sampling before and after surgical treatment to detect recurrence of endometriosis using direct mass spectrometry method. Results: recurrences of EGE were diagnosed in 22% of patients at 18–36 months postoperatively. High incidence of recurrent EGE was reported in the group of patients who did not receive post-surgical hormonal therapy. The hormone therapy with dienogest 2 mg for 6-9 months, as well as with goserelin 3.6 mg for 3-6 months showed the greatest efficacy for the prevention of recurrent EGE. Significant predictive factors of recurrence after surgical treatment for EGE include intensive acyclic pelvic pain; presence of surgical interventions for the history of EGE; infertility; stimulation of ovulation with hormonal drugs as part of the Assisted Reproductive Technologies Program; degree 3-4 disease spread; pronounced adhesions of the small pelvis; infiltrative forms of endometriosis; endometrioid cysts of large size; bilateral ovarian lesions. The plasma lipid profile including determination of phosphatidylcholines, phosphoethanolamines, diglyceride and sphingomyelin in the patients with endometriosis provides an approach for noninvasive early diagnosis of recurrence of this disease by mass spectrometry (sensitivity 93%, specificity 95%). Conclusion: This study developed an individual comprehensive approach to the prevention of recurrent EGE, including surgical treatment, hormonal therapy and diagnosis of the disease at an early stage of development. Timely administration of a-GnRH or dienogest 2 mg at the post-surgery stage is associated with a minimal or total absence of recurrent EGE. The administration of COCs showed no efficacy in preventing the recurrence of EGE, and therefore COCs cannot be recommended as a prophylactic drug, but exclusively for contraceptive use.

Prussian Blue analogue derived porous NiFe2O4 nanocubes for low-concentration acetone sensing at low working temperature

Real-time detection of acetone vapor at low concentration plays a decisive role in early noninvasive diagnosis of diabetes. In this work, porous NiFe2O4 crystalline nanocubes have been scalably prepared via an as-developed cost-efficient and facile strategy, which involves a morphology-inherited annealing treatment of single-resource Prussian Blue analogue of Ni3[Fe(CN)6]2·xH2O solid nanocubes as self-sacrificial templates. The porous NiFe2O4 crystalline nanocubes are demonstrated to be composed of primary nano building blocks and interconnected pores. When utilized as sensing materials, the as-synthesized NiFe2O4 exhibited p-type gas-sensing behavior that the resistance increases in a reducing gas atmosphere. Furthermore, the as-fabricated NiFe2O4 sensor was sensitive and selective to acetone gas with an obvious response value of 1.9 at its low concentration (1 ppm) and low detection of limit (0.52 ppm) at a quite low working temperature (160 °C). In addition, the sensing mechanism is deeply investigated. More significantly, our prominent findings herein shed light on the fabrication of metal oxide based gas sensors in environmental and medicinal fields.

Usefulness of 99mTc-dimercaptosuccinic acid renal scan in the diagnosis and follow-up of acute tubulointerstitial nephritis in children.

BackgroundSymptoms and signs of acute tubulointerstitial nephritis (ATIN) are nonspecific; therefore, renal biopsy is often necessary to clarify the diagnosis. The aim of this study was to evaluate the use of 99mTc-dimercaptosuccinic acid (DMSA) scintigraphy in the diagnosis and follow-up of ATIN.MethodsWe retrospectively reviewed the charts of five patients (nine renal units) with a median age of 14 years who underwent DMSA scan after a clinical and/or biopsy-proven diagnosis of ATIN. The exam was performed within 1 month after disease onset and repeated at a median time of 12 months after the acute phase.ResultsDMSA renal scans performed during the acute phase allowed the discovery of suggestive findings, including diffuse reduction of the renal uptake of radionuclide and presence of multiple ‘cold’ focal lesions in a corticomedullary distribution. The follow-up scintigraphy resulted normal in two patients who were treated with steroids and in one patient who presented a mild renal dysfunction in the acute phase. By contrast, the control scan showed persistent renal damage in one patient who was further readmitted because of hypertension and in one renal transplanted patient who presented a Stage 3 acute kidney injury in the acute phase.ConclusionsDMSA renal scan might be a reliable tool for an early non-invasive diagnosis of ATIN in children and might be particularly useful in those patients who are not candidates for a kidney biopsy. Moreover, DMSA scan gives accurate follow-up evaluation, as it allows monitoring of the evolution of acute renal parenchymal inflammation with potential risk of renal scar formation. Due to the small sample size, our findings warrant further validation in a larger study.

Podocytes regulate the glomerular basement membrane protein nephronectin by means of miR-378a-3p in glomerular diseases

The pathophysiology of many proteinuric kidney diseases is poorly understood, and microRNAs (miRs) regulation of these diseases has been largely unexplored. Here, we tested whether miR-378a-3p is a novel regulator of glomerular diseases. MiR-378a-3p has two predicted targets relevant to glomerular function, the glomerular basement membrane matrix component, nephronectin (NPNT), and vascular endothelial growth factor VEGF-A. In zebrafish (Danio rerio), miR-378a-3p mimic injection or npnt knockdown by a morpholino oligomer caused an identical phenotype consisting of edema, proteinuria, podocyte effacement, and widening of the glomerular basement membrane in the lamina rara interna. Zebrafish vegf-A protein could not rescue this phenotype. However, mouse Npnt constructs containing a mutated 3'UTR region prevented the phenotype caused by miR-378a-3p mimic injection. Overexpression of miR-378a-3p in mice confirmed glomerular dysfunction in a mammalian model. Biopsies from patients with focal segmental glomerulosclerosis and membranous nephropathy had increased miR-378a-3p expression and reduced glomerular levels of NPNT. Thus, miR-378a-3p-mediated suppression of the glomerular matrix protein NPNT is a novel mechanism for proteinuria development in active glomerular diseases.

Raman Spectrometric Detection Methods for Early and Non-Invasive Diagnosis ofAlzheimer's Disease.

The continuous increasing rate of patients suffering of Alzheimer's disease (AD) worldwide requires the adoption of novel techniques for non-invasive early diagnosis and monitoring of the disease. Here we review the various Raman spectroscopic techniques, including Fourier Transform-Raman spectroscopy, surface-enhanced Raman scattering spectroscopy, coherent anti-Stokes Raman scattering spectroscopy, and confocal Raman microspectroscopy, that could be used for the diagnosis of AD. These techniques have shown the potential to detect AD biomarkers, such as the amyloid-β peptide and the tau protein, or the neurotransmitters involved in the disease (e.g., Glutamate and γ-Aminobutyric acid), or the typical structural alterations in specific brain areas. The possibility to detect the specific biomarkers in liquid biopsies and to obtain high resolution 3D microscope images of the affected area make the Raman spectroscopy a valuable ally in the early diagnosis and monitoring of AD.

Expression profiling of maternal plasma and placenta microRNAs in preeclamptic pregnancies by microarray technology

Preeclampsia (PE) is one of the leading causes of maternal and fetal morbidity and mortality, occurring usually in the second half of pregnancy and affecting approximately 5–8% of pregnancies in the world. miRNAs play critical role in the regulation of placental development processes. We aimed to determine specific novel miRNAs for early diagnosis of preeclampsia which is one of the most dangerous pregnancy diseases. In this study 72 samples, maternal age 22 ≤ and ≤36, have been analyzed; maternal plasma and placental miRNAs were isolated from 18 severe preeclampsia (sPE) patients and 18 controls, respectively. Profiling of human miRNAs (1368 probe) was performed in samples with Agilent v16 microarrays for detection of the differences in miRNA expression between two groups. The results were validated by using TaqMan RT-qPCR method. The analysis indicated that 406 of these miRNAs in all placentas and 42 of these miRNAs in all maternal plasma were expressed. The relative expression analysis has shown that 12 miRNAs (p < 0.05 and >2-fold) in maternal plasma were differentially expressed in PE and control group. However, five miRNAs were validated by qRT-PCR. Once validated miRNAs have been searched in databases for their target genes and function, it has been shown that there are some preeclampsia related pathways as a target such as angiogenesis, cardiovascular, hypertension, placental abruption and preeclampsia disorders. Differentially expressed and validated plasma miRNAs might be used as notable biomarkers for non-invasive early diagnosis of preeclampsia and treatment of disease.

Power of screening tests for colorectal cancer enhanced by high levels of M2-PK in addition to FOBT

Colorectal cancer (CRC) is a multistep process that involves adenoma-carcinoma sequence. CRC can be prevented by routine screening, which can detect precancerous lesions. The aim of this study is to clarify whether faecal occult blood test (i-FOBT), tumor M2 pyruvate kinase (t-M2-PK), and endocannabinoid system molecules (cannabinoid receptors type 1-CB1, type 2-CB2, and fatty acid amide hydrolase-FAAH) might represent better diagnostic tools, alone or in combination, for an early diagnosis of CRC. An immunochemical FOB test (i-FOBT) and quantitative ELISA stool test for t-M2-PK were performed in 127 consecutive patients during a 12month period. Endocannabinoid system molecules and t-M2-PK expression were detected by immunostaining in healthy tissues and normal mucosa surrounding adenomatous and cancerous colon lesions. i-FOBT and t-M2-PK combination leads to a better diagnostic accuracy for pre-neoplastic and neoplastic colon lesions. T-M2-PK quantification in stool samples and in biopsy samples (immunostaining) correlates with tumourigenesis stages. CB1 and CB2 are well expressed in healthy tissues, and their expression decreases in the presence of advanced stages of carcinogenesis and disappears in CRC. FAAH signal is well expressed in normal mucosa and low-risk adenoma, and increased in high-risk adenoma and carcinoma adjacent tissues. This study shows that high levels of t-M2-PK in addition to FOBT enhance the power of a CRC screening test. Endocannabinoid system molecule expression correlates with colon carcinogenesis stages. Developing future faecal tests for their quantification must be undertaken to obtain a more accurate early non-invasive diagnosis for CRC.

1-Butyl-3-Methylimidazolium Tetrafluoroborate Film as a Highly Selective Sensing Material for Non-Invasive Detection of Acetone Using a Quartz Crystal Microbalance.

Breath acetone serves as a biomarker for diabetes. This article reports 1-butyl-3-methylimidazolium tetrafluoroborate ([bmim][BF4]), a type of room temperature ionic liquid (RTIL), as a selective sensing material for acetone. The RTIL sensing layer was coated on a quartz crystal microbalance (QCM) for detection. The sensing mechanism is based on a decrease in viscosity and density of the [bmim][BF4] film due to the solubilization of acetone leading to a positive frequency shift in the QCM. Acetone was detected with a linear range from 7.05 to 750 ppmv. Sensitivity and limit of detection were found to be 3.49 Hz/ppmv and 5.0 ppmv, respectively. The [bmim][BF4]-modified QCM sensor demonstrated anti-interference ability to commonly found volatile organic compounds in breath, e.g., isoprene, 1,2-pentadiene, d-limonene, and dl-limonene. This technology is useful for applications in non-invasive early diabetic diagnosis.

Exosomal surface protein markers in diagnosis of colorectal cancer

Colorectal cancer (CRC) is one of the most common primary malignancies. Early stages of the disease are asymptomatic in the majority of cases, leading to late detection and high mortality. Available noninvasive diagnostic techniques are limited in sensitivity and specificity, and designing new ones is still a pressing problem. Exosomes are membrane-derived microvesicles secreted into human biological fluids and provide a novel way to assess the course of an oncology disease. The review describes the repertoire of exosomal surface biomarkers found in the blood of CRC patients and the prospects of employing multiplexed tests for exosomal markers in early noninvasive diagnosis of cancer.

A novel non-invasive detection method for the FGFR3 gene mutation in maternal plasma for a fetal achondroplasia diagnosis based on signal amplification by hemin-MOFs/PtNPs

The small amount of cell-free fetal DNA (cffDNA) can be a useful biomarker for early non-invasive prenatal diagnosis (NIPD) of achondroplasia. In this study, a novel non-invasive electrochemical DNA sensor for ultrasensitive detecting FGFR3 mutation gene, a pathogenic gene of achondroplasia, based on biocatalytic signal materials and the biotin-streptavidin system are presented. Notably encapsulation of hemin in metal-organic frameworks-based materials (hemin-MOFs) and platinum nanoparticles (PtNPs) were used to prepare hemin-MOFs/PtNPs composites via a one-beaker-one-step reduction. We utilized hemin-MOFs/PtNPs for signal amplification because the promising hemin-MOFs/PtNPs nanomaterial has remarkable ability of catalyze H2O2 as well as excellent conductivity. To further amplify the electrochemical signal, reduced graphene oxide-tetraethylene pentamine (rGO-TEPA), gold nanoparticles and streptavidin were selected for modification of the electrode to enhance the conductivity and immobilize more biotin-modified capture probe (Bio-CP) through the high specificity and superior affinity between streptavidin and biotin. The electrochemical signal was primarily derived from the synergistic catalysis of H2O2 by hemin and PtNPs and recorded by Chronoamperometry. Under the optimal conditions, this newly designed biosensor exhibited sensitive detection of FGFR3 from 0.1fM to 1nM with a low detection limit of 0.033fM (S/N=3). We proposed that this ultrasensitive biosensor is useful for the early non-invasive prenatal diagnosis of achondroplasia.

Targeted Metabolic Profiling of the Tg197 Mouse Model Reveals Itaconic Acid as a Marker of Rheumatoid Arthritis.

Rheumatoid arthritis is a progressive, highly debilitating disease where early diagnosis, enabling rapid clinical intervention, would provide obvious benefits to patients, healthcare systems, and society. Novel biomarkers that enable noninvasive early diagnosis of the onset and progression of the disease provide one route to achieving this goal. Here a metabolic profiling method has been applied to investigate disease development in the Tg197 arthritis mouse model. Hind limb extract profiling demonstrated clear differences in metabolic phenotypes between control (wild type) and Tg197 transgenic mice and highlighted raised concentrations of itaconic acid as a potential marker of the disease. These changes in itaconic acid concentrations were moderated or indeed reversed when the Tg197 mice were treated with the anti-hTNF biologic infliximab (10 mg/kg twice weekly for 6 weeks). Further in vitro studies on synovial fibroblasts obtained from healthy wild-type, arthritic Tg197, and infliximab-treated Tg197 transgenic mice confirmed the association of itaconic acid with rheumatoid arthritis and disease-moderating drug effects. Preliminary indications of the potential value of itaconic acid as a translational biomarker were obtained when studies on K4IM human fibroblasts treated with hTNF showed an increase in the concentrations of this metabolite.