Detection Of New Biomarkers Research Articles

Urinary Smad1 as a New Biomarker for Diabetic Nephropathy in Patients with Type 1 Diabetes Mellitus

Introduction: Type 1 diabetes mellitus is an autoimmune disease affecting children worldwide with many consequences disturbing their quality of life. Diabetic nephropathy is among the most important complications of T1DM. The reliability of albuminuria as a predictor and prognosticator for renal injury has been frequently questioned. Many markers were investigated to replace urinary albumin. The current study aimed to assess the potential value of urinary Smad1 as a new biomarker for the early diagnosis and assessment of severity of diabetic nephropathy in children with. Material and method: A case control cross sectional study with 53 patients with type 1 diabetes mellitus. 30 patients with diabetic nephropathy including 19 patients with microalbuminuria (urinary albumin creatinine ratio: 30-300 mg/gm) and 11 patients with macroalbuminuria (urinary albumin creatinine ratio: >300 mg/gm). The remainder 23 patients had normal urinary albumin levels. In addition, there were 20 healthy age and sex matched children who served as control group. In all subjects we assessed urinary albumin, urinary creatinine and urinary albumin creatinine ratio and urinary Smad1 levels Results: Urinary Smad1 levels and urinary Smad1 creatinine ratio show good sensitivity and specificity in detection of DN as compared to urinary albumin. The performance of SCR was better than urinary Smad1 with sensitivity and specificity of 100.0% and 96.0% for SCR, compared to sensitivity and specificity of 90.0% and 91.0% for urinary Smad1. Conclusion: Urinary Smad1 is a promising new biomarker for detection of diabetic nephropathy in patients with type 1 diabetes mellitus with high sensitivity and specificity.

Differential proteins among normal cervix cells and cervical cancer cells with HPV-16 infection, through mass spectrometry-based Proteomics (2D-DIGE) in women from Southern México.

BackgroundCervical cancer (CC) is the fourth most common cancer in women worldwide with an estimated 528,000 new cases in 2012. The same year México had an incidence of 13,960 and a mortality of 4769 cases. There are several diagnosis methods of CC; among the most frequents are the conventional Pap cytology (Pap), colposcopy, and visual inspection with acetic acid (VIA), histopathological examination, tests of imaging and detection of high-risk papilloma virus (HR-HPV) with molecular tests (PCR, hybridization, sequencing). Proteomics is a tool for the detection of new biomarkers that can be associated with clinical stage, histological type, prognosis, and/or response to treatment. In this study we performed a comparative analysis of CC cells with normal cervical cells. The proteomic analysis was carried out with the fluorescent two-dimensional electrophoresis (2D-DIGE) technique to subsequently identify differential protein profiles using Decyder Software, and the selected proteins were identified by Mass Spectrometry (MALDI-TOF).ResultsThe proteins that showed an increased expression in cervical cancer in comparison with normal cervix cells were: Mimecan, Actin from aortic smooth muscle and Lumican. While Keratin, type II cytoskeletal 5, Peroxiredoxin-1 and 14-3-3 protein sigma showed a decrease in their protein expression level in cervical cancer in comparison with normal cervix cells.ConclusionsThus, this study was successful in identifying biomarker signatures for cervical cancer, and might provide new insights into the mechanism of CC progression.

Next Generation Sequencing: A Tool for This Generation of Nephrologists

The emergence of next generation sequencing (NGS) techniques has made the sequencing of whole genomes, transcriptomes, and epigenomes faster and more readily available than previous methods such as Sanger sequencing, which was developed in the 1970s. It is now 10 years since NGS began to revolutionise biological and medical research. Sequencing of RNA provides insights into up or downregulated gene expression patterns and therefore into molecular disease mechanisms. This can lead to the detection of new biomarkers that can be used as diagnostic tools in risk stratification, or even as new therapeutic targets. In nephrology, NGS plays a role in both basic and experimental research, but also in the clinical setting, whereby the diagnosis of innate genetic diseases such as ciliopathies or genetically moderated acquired diseases such as glomerulopathies has improved. NGS enables precise diagnosis and classification of common diseases of the kidneys and urinary tract, aids in both prognostic and predictive decision-making, and in the avoidance of unnecessary therapies. It also plays a role in the risk stratification of disease recurrence after transplantation. NGS is a robust method; however, the performance of NGS is dependent on the method of tissue storage, the extraction of DNA or RNA, and on the sequencing platform itself, as well as on the bioinformatic analyses performed, integration of clinical data, and comprehensive interpretation of the results. The aim of this article is to review and emphasise the importance of NGS as a tool for this generation of nephrologists.

Hepatic metastatic niche: from normal to pre-metastatic and metastatic niche.

Liver is the organ responsible for hematopoiesis during fetal life, which is also a target organ of metastasis for several cancers. In order to recognize the hepatic metastatic changes, obtain a better grasp of cancer prevention, treatment, and inhibition mode of hepatic metastasis progression, we investigate the changes and transformation of normal hepatic niche cells to metastatic niche ones in this review. On the other hand, since metastatic diseases alter the liver function, the changes in a number of cancers that metastasize to the liver have also been reviewed. Relevant English-language literature was searched and retrieved from PubMed (1994-2014) using the following keywords: hepatic stem cell niche, hepatic metastatic niche, chemokine, and microRNAs (miRNAs). Also, over 86 published studies were investigated, and bioinformatics analysis of differentially expressed miRNAs in hepatic cancer and metastasis was performed. Metastasis is developed in several stages with specific changes and mechanisms in each stage. Recognition of these changes would lead to detection of new biomarkers and clinical targets involved in specific stages of liver metastasis. Investigation of the hepatic stem cell niche, development of metastasis in liver tissue, as well as changes in chemokines and miRNAs in metastatic hepatic niche can significantly contribute to faster detection of liver metastasis progression.

Autologous anti-Aß antibodies in CAA-ri: New biomarker for detection of amyloid-related imaging abnormalities (ARIA) during Aß-disease modifying therapies for AD

Background: Cerebral Amyloid Angiopathy-related inflammation (CAA-ri) is a rare disease characterized by vasogenic edema and multiple cortical/subcortical microbleeds, shearing several aspects with the recently defined Amyloid-Related Imaging Abnormalities (ARIA) during Alzheimer'/INS;s disease (AD) passive immunization therapies.

Immune Profiling in Blood Identifies sTNF-R1 Performing Comparably Well as Biomarker Panels for Classification of Alzheimer's Disease Patients

Alzheimer's disease (AD) has been linked to a state of cerebral and systemic inflammation. The objective of the present study was to determine whether singular markers or a set of inflammatory biomarkers in peripheral blood allow discrimination between AD patients and healthy controls at the individual level. Using bead based multiplexed sandwich immunoassays, 25 inflammatory biomarkers were measured in 164 serum samples from individuals with early AD and age-matched cognitively healthy elderly controls. The data set was randomly split into a training set for feature selection and classification training and a test set for class prediction of blinded samples (1 : 1 ratio) to evaluate the chosen predictors and parameters. Multivariate data analysis was performed with use of a support vector machine (SVM). After selection of sTNF-R1 as most discriminative parameter in the training set, the application of SVM to the independent test dataset resulted in a 90.0% correct classification for individual AD and control subjects. We identified sTNF-R1 from a marker set consisting of 25 inflammatory biomarkers, which allowed SVM-based discrimination of AD patients from healthy controls on a single-subject classification level comparably well as biomarker panels with a clinically relevant accuracy and validity. Although larger sample populations will be needed to confirm this diagnostic accuracy, our study suggests that sTNF-R1 in serum-either as singular marker or incorporated into a biomarker panel-could be a powerful new biomarker for detection of AD. In addition, selective inhibition of TNF-R1 function may represent a new therapeutic approach in AD.

Signal Transduction Alterations in Glioma: Implications for Diagnosis and Therapy

Malignant gliomas are the leading cause of central-nervous-system-tumour-related death, and despite recent advances in surgery, radiotherapy, and chemotherapy, current treatment regimens have a modest survival benefit; the prognosis is even worse in children with brain stem malignant gliomas. Gliomas are divided into four clinical grades on the basis of their histology and prognosis. The most malignant grade 4 astrocytoma, or glioblastoma multiforme (GBM), either arises de novo (usually associated to epidermal growth factor receptor pathway activation and PTEN inactivation leading to PI3K kinase/AKT activation pathway) or progresses from lower grade to higher grade over time (characteristically due to p53 and retinoblastoma pathways inactivation). Due to a combination of its complex phenotype and organ-specific clinical manifestations, efforts to refine GBM treatment with targeted therapies have largely been frustrated. Therefore, the identification and characterization of signal transduction pathways alterations, with a pathogenic role on glioma development and progression, may contribute to the identification of therapeutic targets aimed at a more efficient treatment. The seven tightly organized papers in this special issue give an update of all the latest concepts about the molecular mechanisms of pathogenesis of glioblastoma and new therapeutic opportunities. The molecular characteristics of angiogenesis, a key event for glioma survival, aggressiveness and growth, are addressed by two well-balanced papers. S. Bulnes et al. review angiogenic signaling altered in a rat glioma model and discuss on the selection mechanisms for more aggressive subpopulation with invasive phenotype. They show that glioma stem cells and vascular endothelial cells play a relevant role in the angiogenic process, and referring to molecular pathways, hypoxia inducible factor-1 and vascular endothelial growth factor are the most significant. The papers by V. Cea et al. offers an overview of the most relevant issues about antiangiogenic therapy for glioma, presenting several available drugs that are used or can potentially be utilized for the inhibition of angiogenesis in glioma, focusing on the key mediators of the molecular mechanisms underlying the resistance of glioma to anti-angiogenic therapy. Two interesting and novel papers discuss epigenetic mechanisms producing signal pathways deregulation in gliomas. The paper by R. Alelu-Paz et al. is a nice addition to the current literature about epigenetic changes in human cancer, particularly in gliomas. The emerging role of cancer stem cells in the pathophysiology of cancer is as well discussed. R. Martinez has written a paper describing epigenetic and genetic alterations in gliomas, resulting in deregulation or functional disruption of tumor suppressor and oncogenes. In both papers, the discussion of epigenetic alterations in the pathogenesis and evolution of gliomas clearly indicate their crucial function for discovering new biomarkers for detection and prognosis and for development of new pharmacological strategies. L. Catacuzzeno et al. clearly introduce the reader to the structural, biophysical, pharmacological, and modulatory properties of the intermediate conductance calcium-activated K (KCa3.1) channels. They describe the importance of the KCa3.1 channels in glioblastoma cell functions. These channels are highly expressed in glioblastoma cells if compared to the normal brain parenchyma and play an important role in the control of glioblastoma cell migration, a critical process that represents major causes for tumor progression and for recurrence following tumor surgical resection. Altogether, data suggest KCa3.1 channels as potential candidates for a targeted therapy against glioma. The research paper by H. L. Watt et al. evaluates the biological responses of glioma cells to combined treatment with RTK inhibitors, DNA damaging agents, and octreotide, an agonist of the somatotropin receptor. Changes in the activation profile of EGFR mitogenic signaling and DNA damage response pathway, as well as apoptosis and cell cycle distribution were analyzed. The results support the notion that the effects of combined therapy on glioma cells mostly depend on the specific context of cell cycle arrest. A crucial challenge for human glioma treatment is to deliver drugs effectively to invasive glioma cells residing in a sanctuary within the central nervous system. S. Catuogno et al. discuss recent results on the use of oligonucleotides that will hopefully provide new effective treatment for gliomas. Oligonucleotide-based approaches, including antisense, microRNAs, small interfering RNAs, and nucleic acid aptamers, look very promising particularly to overcome challenges presented by the blood-brain barrier. In total, we hope that these contributions will provide a well-rounded overview of histopathology, molecular biology, and current treatment strategies for glioma.

Advances in biomarker research for pancreatic cancer.

Pancreatic cancer (PC) is a leading cause of cancer related deaths in United States. The lack of early symptoms results in latestage detection and a high mortality rate. Currently, the only potentially curative approach for PC is surgical resection, which is often unsuccessful because the invasive and metastatic nature of the tumor masses makes their complete removal difficult. Consequently, patients suffer relapses from remaining cancer stem cells or drug resistance that eventually lead to death. To improve the survival rate, the early detection of PC is critical. Current biomarker research in PC indicates that a serum carbohydrate antigen, CA 19-9, is the only available biomarker with approximately 90% specificity to PC. However, the efficacy of CA 19-9 for assessing prognosis and monitoring patients with PC remains contentious. Thus, advances in technology and the detection of new biomarkers with high specificity to PC are needed to reduce the mortality rate of pancreatic cancer.

Comparison of different liquid chromatography stationary phases in LC‐HRMS metabolomics for the detection of recombinant growth hormone doping control

Growth hormone (GH) is a polypeptide suspected of being used in horse racing to speed up physical performances. Despite scientific advances in the recent years, the control of its administration remains difficult. In order to improve it, a metabolomics study through LC-high resolution mass spectrometry measurements was recently initiated to assess the metabolic perturbations caused by recombinant equine growth hormone administration. Few tens of ions not identified structurally were highlighted as compounds responsible for the modification of metabolic profiling observed in treated animals. This previous work was based on the use of Uptisphere Strategy NEC as the chromatographic column. In parallel, more and more metabolomics studies showed the interest of the use of new chromatographic supports such as hydrophilic interaction chromatography for the analysis of polar compounds. It is in this context that an investigation was conducted on Uptisphere HDO and Luna hydrophilic interaction chromatography stationary phases to generate and process urinary metabolomics fingerprints, which could allow to establish a comparison with Uptisphere Strategy NEC. The chromatographic column the most adapted for the detection of new biomarkers of GH administration has been used to set up a relevant statistical model based on the analysis of more than hundred biological samples.

Abstract 4861: Identification of premalignant bladder markers using DNA microarray analysis of the UPII-SV40Tag model for invasive bladder UCC

Abstract Urothelial cell carcinoma of the bladder (UCC) ranks 5th in incidence of all cancers in the developed world, yet the mechanisms of its origin and progression remain poorly understood. A few urine-based tests for screening and surveillance are commercially available for bladder cancer, but there are no reliable non-invasive methods for detecting premalignant UCC. In order to determine molecular mechanisms involved in early UCC development and to identify new biomarkers for detection, diagnosis and prognosis of UCC, a transgenic mouse model for invasive bladder cancer (UPII-SV40Tag mice) was analyzed by DNA microarray technology. Genes that are differentially expressed between the bladders of UPII-SV40Tag mice and their age-matched wild type (WT) littermates at 3, 6, 20 and 30 weeks of age were identified. The ages selected correspond to premalignant, carcinoma in-situ, and early and later stage papillary UCC, respectively. Approximately 1,900 unique genes were differentially expressed (≥ 3-fold difference at one or more time points) between WT and UPII-SV40Tag urothelium during the time course of tumor development. A high proportion of cell cycle regulatory genes and proliferation signaling genes were more strongly expressed in the UPII-SV40Tag bladder urothelium. Several of the genes upregulated and downregulated in UPII-SV40Tag urothelium were tested as biomarkers for premalignancy, including autocrine motility factor (AMFR), hyaluronan-mediated motility receptor (Hmmr), proliferating cell nuclear antigen (PCNA), Rac GTPase activating protein 1 (RacGAP1), superoxide dismutase 3 (SOD3), collagen 1a2 (Col1a2), and others. These tests were performed by analyzing urine samples from a recently completed Phase II, randomized, placebo controlled chemoprevention trial (N01 CN85186, PI: A. Sabichi) that was designed to test whether celecoxib can prevent recurrence in patients successfully treated by transurethral resection (TUR) for non-muscle invasive bladder cancer. Positive markers, Hmmr and RacGAP1, and a negative marker, Col1a2, have been identified in patient urine samples that show promise for detecting UCC recurrence. We are currently attempting to determine the biological function of Hmmr, and RacGAP1 in bladder premalignancy. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4861. doi:10.1158/1538-7445.AM2011-4861

Blood-based diagnostics of traumatic brain injuries

Traumatic brain injury is a major health and socioeconomic problem that affects all societies. However, traditional approaches to the classification of clinical severity are the subject of debate and are being supplemented with structural and functional neuroimaging, as the need for biomarkers that reflect elements of the pathogenetic process is widely recognized. Basic science research and developments in the field of proteomics have greatly advanced our knowledge of the mechanisms involved in damage and have led to the discovery and rapid detection of new biomarkers that were not available previously. However, translating this research for patients’ benefits remains a challenge. In this article, we summarize new developments, current knowledge and controversies, focusing on the potential role of these biomarkers as diagnostic, prognostic and monitoring tools of brain-injured patients.

Identification of genes correlated with early-stage bladder cancer progression.

Transitional cell carcinoma (TCC) of the bladder ranks fourth in incidence of all cancers in the developed world, yet the mechanisms of its origin and progression remain poorly understood. There are also few useful diagnostic or prognostic biomarkers for this disease. We have combined a transgenic mouse model for invasive bladder cancer (UPII-SV40Tag mice) with DNA microarray technology to determine molecular mechanisms involved in early TCC development and to identify new biomarkers for detection, diagnosis, and prognosis of TCC. We have identified genes that are differentially expressed between the bladders of UPII-SV40Tag mice and their age-matched wild-type littermates at 3, 6, 20, and 30 weeks of age. These are ages that correspond to premalignant, carcinoma in situ, and early-stage and later stage invasive TCC, respectively. Our preliminary analysis of the microarray data sets has revealed approximately 1,900 unique genes differentially expressed (> or =3-fold difference at one or more time points) between wild-type and UPII-SV40Tag urothelium during the time course of tumor development. Among these, there were a high proportion of cell cycle regulatory genes and a proliferation signaling genes that are more strongly expressed in the UPII-SV40Tag bladder urothelium. We show that several of the genes upregulated in UPII-SV40Tag urothelium, including RacGAP1, PCNA, and Hmmr, are expressed at high levels in superficial bladder TCC patient samples. These findings provide insight into the earliest events in the development of bladder TCC as well as identify several promising early-stage biomarkers.

Abstract A16: DNA microarray analysis of the UPII-SV40Tag model for invasive bladder TCC: Identification of markers for bladder premalignancy

Abstract Transitional cell carcinoma of the bladder (TCC) ranks 4th in incidence of all cancers in the developed world, yet the mechanisms of its origin and progression remain poorly understood. There are a few commercially available urine-based tests for screening and surveillance for bladder cancer, but none of these can detect premalignancy. We have combined a transgenic mouse model for invasive bladder cancer (UPII-SV40Tag mice) with DNA microarray technology, in order to determine molecular mechanisms involved in early TCC development and to identify new biomarkers for detection, diagnosis and prognosis of TCC. We have identified genes that are differentially expressed between the bladders of UPII-SV40Tag mice and their age-matched wild type (WT) littermates at 3, 6, 20 and 30 weeks of age. These are ages which correspond to premalignant, carcinoma in-situ, and early and later stage papillary TCC, respectively. There were approximately 1,900 unique genes differentially expressed (>= 3-fold difference at one or more time points) between WT and UPII-SV40Tag urothelium during the time course of tumor development. Among these, there were a high proportion of cell cycle regulatory genes and proliferation signaling genes that are more strongly expressed in the UPII-SV40Tag bladder urothelium. We have tested several of the genes upregulated in UPII-SV40Tag urothelium, including autocrine motility factor (AMFR), hyaluronan-mediated motility receptor (Hmmr), proliferating cell nuclear antigen (PCNA), Rac GTPase activating protein 1 (RacGAP 1) and others as biomarkers for premalignancy, in urine samples from a recently completed Phase II, randomized, placebo controlled chemoprevention trial (N01 CN85186, PI: A. Sabichi) that was designed to test whether celecoxib can prevent recurrence in patients successfully treated by TUR for non-muscle invasive bladder cancer. Citation Information: Cancer Prev Res 2010;3(1 Suppl):A16.

Glycogen phosphorylase BB could be a new biomarker for detection of cardiac toxicity during hematopoietic cell transplantation for hematological malignancies

Glycogen phosphorylase BB could be a new biomarker for detection of cardiac toxicity during hematopoietic cell transplantation for hematological malignancies

Identification of two new serum protein profiles for renal cell carcinoma

Renal cell carcinoma (RCC) is the most lethal urological cancer, and survival greatly depends on early diagnosis. Therefore, reliable, new biomarkers for detection of RCC are required. We assessed serum protein profiles of samples from two institutes with SELDI-TOF MS in duplicate on CM10 chips at pH 6.0 (set 1: 37 RCC + 32 healthy controls (HC), set 2: 20 RCC + 25 HC). Mean peak intensities of detected proteins were compared between RCC and HC with non-parametric testing. Classification trees were built with discriminating peaks using one sample set as training set and the other as independent validation set. We found 15 peaks significantly different (p<0.01) between RCC and HC. Two classification trees could be built with these peaks. Tree A achieved 75% sensitivity and 85% specificity for cross-validation and 76 and 65% for independent validation. Tree B had 71 and 62% sensitivity and specificity for cross-validation and 83 and 82% for independent validation. Although two serum protein profiles comprising 5 protein peaks were found that could separate RCC from HC, the sensitivity and specificity is not sufficient to recommend large scale use. Upon structural identification and quantitative validation, however, these proteins might prove suitable markers in the follow up of RCC patients.

Role of Osteopontin and CD44s Expression for Patients With Hepatocellular Carcinoma Undergoing Liver Transplantation or Resection

Background Detection of new biomarkers for hepatocellular carcinoma (HCC) is needed to estimate prognosis after liver transplantation (OLT) or hepatic resection. Osteopontin (OPN) is a secreted, calcium-binding, phosphorylated, acidic glycoprotein that is overexpressed in various cancers. Cluster differentiation 44 standard isoform (CD44s) is one of the primary receptors of OPN; it may contribute to metastatic tumor spread. Materials and Methods Tumor tissue and surrounding hepatic parenchyma were obtained from 53 HCC patients who underwent liver resection. Their RNA was extracted from nitrogen-frozen tissues, and OPN mRNA levels were estimated by quantitative reverse transcription–polymerase chain reactions. Formalin-fixed, paraffin-embedded tissues were obtained from the same patients, and additionally from 60 OLT HCC patients to perform expression analysis for OPN and CD44s by standard avidin-biotin immunostaining methods. Results Expression of OPN and CD44s was significantly higher among HCC compared with adjacent nontumor tissue. The OPN mRNA expression and protein abundance correlated positively; OPN overexpression was associated with high tumor grade. A positive correlation existed between OPN and CD44s expression; both proteins were significantly overexpressed in HCC lesions with positive lymph nodes. No significant correlation existed between patient survival and OPN and CD44s expression. Conclusion Expression of both OPN and CD44s in HCC is associated with advanced tumor stage, thus possibly contributing prognostic information when evaluated together with classical clinicopathological parameters.

Abstract B52: Identification of genes involved in early stage bladder cancer progression.

Abstract B52 Transitional cell carcinoma of the bladder (TCC) ranks 4th in incidence of all cancers in the developed world, yet the mechanisms of its origin and progression remain poorly understood. There are also few useful diagnostic or prognostic biomarkers for this disease. We have combined a transgenic mouse model for invasive bladder cancer (UPII-SV40T mice) with DNA microarray technology, in order to determine molecular mechanisms involved in early TCC development and to identify new biomarkers for detection, diagnosis and prognosis of TCC. We have identified genes that are differentially expressed between the bladders of UPII-SV40T mice and their age-matched wild type (WT) littermates at 3, 6, 20 and 30 weeks of age. These are times which correspond to premalignant, carcinoma in-situ, and early and later stage papillary TCC, respectively. There were approximately 1,900 unique genes differentially expressed (≥ 3-fold difference at one or more time points) between WT and UPII-SV40T urothelium during the time course of tumor development. Among these, there were a high proportion of cell cycle regulatory genes and cytokinesis genes that are more strongly expressed in the UPII-SV40T bladder urothelium. We have tested several of the genes upregulated in UPII-SV40T urothelium, including autocrine motility factor (AMFR), Caspase 3, Cox-2, PCNA and others as biomarkers for premalignancy in urine samples from a completed chemoprevention trial. These results will be discussed in the context of the UPII-SV40T model. Citation Information: Cancer Prev Res 2008;1(7 Suppl):B52.

Use of biomarkers for diagnosis and management of traumatic brain injury patients

Advances in the understanding of human biochemistry and physiology have provided insight into new pathways by which we can understand traumatic brain injury (TBI). Increased sophistication of laboratory techniques and developments in the field of proteomics has led to the discovery and rapid detection of new biomarkers not previously available. To review recent advances in biomarker research for traumatic brain injury, describe the features of the ideal biomarker and to explore the potential role of these biomarkers in improving clinical management of brain injured patients. Through a literature review of recent research on TBI biomarkers and through experience with TBI research, important elements of biomarker development are described together with potential applications to patient care. TBI biomarkers could have a significant impact on patient care by assisting in the diagnosis, risk stratification and management of TBI. Biomarkers could provide major opportunities for the conduct of clinical research, including confirmation of injury mechanism(s) and drug target identification. Continuing studies by the authors' group are now being conducted to elucidate more fully the relationships between new biomarkers and severity of injury and clinical outcomes in all severities of TBI patients.

Potential biomarkers for diagnosis of sarcoidosis using proteomics in serum

Sarcoidosis is a multi-systemic inflammatory disorder, which affects the lungs in 90% of the cases. The main pathologic feature is chronic inflammation resulting in non-caseating granuloma formation. Until now there is no satisfying biomarker for diagnosis or prognosis of sarcoidosis. This study is focused on the detection of potential biomarkers in serum for the diagnosis of sarcoidosis using surface-enhanced laser desorption ionization-time of flight-mass spectrometry (SELDI-TOF-MS). For detection of potential biomarkers, protein profiles of anion exchange fractionated serum of 35 sarcoidosis patients and 35 healthy controls were compared using SELDI-TOF-MS. Sensitivities and specificities of the potential biomarkers obtained with SELDI-TOF-MS, generated with decision tree algorithm, were compared to the conventional markers angiotensin converting enzyme (ACE) and soluble interleukin-2 receptor (sIL-2R). Optimal classification was achieved with metal affinity binding arrays. A single marker with a mass-to-charge (m/z) value of 11,955 resulted in a sensitivity and specificity of 86% and 63%, respectively. A multimarker approach of two peaks, m/z values of 11,734 and 17,377, resulted in a sensitivity and specificity of 74% and 71%, respectively. These sensitivities and specificities were higher compared to measurements of ACE and sIL-2R. Identification of the peak at m/z 17,377 resulted in the alpha-2chain of haptoglobin. This study acts as a proof-of-principle for the use of SELDI-TOF-MS in the detection of new biomarkers for sarcoidosis. The peak of the multimarker at m/z 17,377 was identified as the alpha-2chain of haptoglobin.

An Integrated Clinical-Genomics Approach Identifies a Candidate Multi-Analyte Blood Test for Serous Ovarian Carcinoma

Cancer of the ovary confers the worst prognosis among women with gynecologic malignancies, underscoring the need to develop new biomarkers for detection of early disease, particularly those that can be readily monitored in the blood. We developed an algorithm to identify secreted proteins encoded among approximately 22,500 genes on commercial oligonucleotide arrays and applied it to gene expression profiles of 67 stage I to IV serous papillary carcinomas and 9 crudely enriched normal ovarian tissues, to identify putative diagnostic markers. ELISAs were used to validate increased levels of secreted proteins in patient sera encoded by genes with differentially high expression. We identified 275 genes predicted to encode secreted proteins with increased/decreased expression in ovarian cancers (<0.5- or >2-fold, P < 0.001). The serum levels of four of these proteins (matrix metalloproteinase-7, osteopontin, secretory leukoprotease inhibitor, and kallikrein 10) were significantly elevated in a series of 67 independent patients with serous ovarian carcinomas compared with 67 healthy controls (P < 0.001, Wilcoxon rank sum test). Optimized support vector machine classifiers with as few as two of these markers (osteopontin or kallikrein 10/matrix metalloproteinase-7) in combination with CA-125 yielded sensitivity and specificity values ranging from 96% to 98.7% and 99.7% to 100%, respectively, with the ability to discern early-stage disease from normal, healthy controls. Our data suggest that this assay combination warrants further investigation as a multi-analyte diagnostic test for serous ovarian adenocarcinoma.