New Marker For Detection Research Articles

DIP-microhaplotypes: new markers for detection of unbalanced DNA mixtures.

The identification of a suspect in a degraded and unbalanced DNA mixture has been a challenge for the standard short tandem repeat polymorphisms (STR) typing. Several methods have been introduced to solve this problem, such as DIP-STR, DIP-SNP, and SNP-STR markers. In this study, we proposed DIP-microhaplotype (deletion/insertion linked a chain of SNPs) as a kind of new genetic marker to type the unbalanced and degraded DNA mixture. We established the detection method with ten DIP-microhaplotype markers including 26 SNPs using allele-specific multiplex PCR followed by SNaPshot assay. This novel compound marker allows us to detect the minor DNA with a sensitivity of 1:100 to 1:1000 in a DNA mixture of any gender. Most of the DIP-microhaplotype markers had a relatively high probability of informative alleles with an average informative value (I value) of 0.308. In all, we proposed DIP-microhaplotype as a novel type of DNA marker for the detection of minor contributor from unbalanced DNA mixtures. Due to their inherent shorter length, higher polymorphism, and sensitivity, DIP-microhaplotypes are promising markers for the examination of the degraded and unbalanced mixtures in forensic stains or clinical chimeras.

Lipid indices in women blood serum before and after the procedure of removal of mature ovarian teratoma

Purpose of Investigation: The aim of this study was to find new markers for detection of ovarian neoplasms. Authors decided to compare the lipid indicies in the blood serum of patients undergoing surgery treatment of mature ovarian teratomas. Lack of data on this subject in accessible literature was a stimulating factor for this study. Materials and Methods: The examinations covered two groups of women, each with 32 persons. Fasting blood serum gained from blood samples was analysed, marking before the surgical procedure, on the 7th day, six and 12 months after the procedure. Correlations between lipid indices changes HDL/TCH, HDL/LDL, and ApoA1/ApoB was analysed with use of Spearman correlation test. Results: In all parameters of the lipid metabolism and lipid indices, similar direction of changes - that is a trend direction, has been observed. Conclusions: Lipid indices showed their usefulness in preoperational monitoring, the effectiveness of treatment, and postoperative monitoring of mature ovarian teratomas.

Subfractional distribution of Apo B-containing lipoproteins: a new marker for detection of individual cardiovascular risk

Subfractional distribution of Apo B-containing lipoproteins: a new marker for detection of individual cardiovascular risk

Abstract 3258: Targeting premalignant lesions for early breast cancer detection and intervention

Abstract Breast cancer patients' outcome and survival depends on the timely diagnosis of early malignant lesions. The lack of molecular understanding of the early changes in breast tissue that facilitate tumor progression has limited the development of tools to non-invasively distinguish early stage disease from normal breast tissue. Therefore, probes to target and better understand early breast tumors are much needed. Our laboratory has pioneered in vivo screening of phage libraries to identify peptides that specifically recognize tumor vessels, including breast cancer vasculature. These peptides have been employed to specifically deliver drugs, diagnostic agents, and nanoparticles to breast tumors. Breast cancer progression constitutes a multistep process through a series of intermediate hyperplastic and neoplastic stages to invasive carcinoma. In this study, we aimed to identify peptides that specifically recognize premalignant lesions in the mammary tissue. To achieve this goal, we utilized the power of phage display to probe hyperplastic lesions associated with premalignant disease in a transgenic MMTV-PyMT animal model. After multiple ex-vivo and in-vivo rounds of selection, we identified a peptide, Prem-1, that on intravenous administration, specifically homed to premalignant mammary lesions. Prem-1 also homed to fully developed breast tumors in the same animal model, suggesting that the putative receptor for Prem-1 is expressed throughout the progression of the disease. Interestingly, Prem-1 did not show any affinity to normal breast tissue. Furthermore, we also identified 2 other candidate peptides that showed significant homing to premalignant lesions with a very different binding pattern as compared to Prem-1. We hypothesized that all three peptides recognize early changes in the breast tissue microenvironment but each bind a different target receptor in the tissue. We are currently investigating these receptors and analyzing their expression in breast cancer progression. Secondly, we are testing the peptides identified herein for the delivery of therapeutic nanoparticles as a mode of early intervention in breast cancer progression. This project utilized the natural environment in the early breast tumor to probe for new markers for detection of early disease. Methods to detect and study early premalignant mammary lesions will expand our understanding of the natural history of the disease, especially the changes in breast tissue that likely lead to tumor development. Hence, the knowledge gained from this study would provide a basis for therapies aimed at suppressing or eradicating premalignant breast lesions. This would be particularly beneficial for women at high-risk of breast cancer based on their genetic predisposition (mutations in BRCA genes). Citation Format: Aman P. Mann, Ramana Kotamraju, Tambet Teesalu, Erkki Ruoslahti. Targeting premalignant lesions for early breast cancer detection and intervention. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3258. doi:10.1158/1538-7445.AM2014-3258

Breast Cancer in Morocco: A Literature Review

In Morocco, breast cancer is the most prevalent cancer in women and a major public health problem. Several Moroccan studies have focused on studying this disease, but more are needed, especially at the genetic and molecular levels. It is therefore interesting to establish the genetic and molecular profile of Moroccan patients with breast cancer. In this paper, we will highlight some pertinent hypotheses that may enhance breast cancer care in Moroccan patients. This review will give a precise description of breast cancer in Morocco and propose some new markers for detection and prediction of breast cancer prognosis.

FOXQ1, a Novel Target of the Wnt Pathway and a New Marker for Activation of Wnt Signaling in Solid Tumors

BackgroundThe forkhead box transcription factor FOXQ1 has been shown to be upregulated in colorectal cancer (CRC) and metastatic breast cancer and involved in tumor development, epithelial-mesenchymal transition and chemoresistance. Yet, its transcriptional regulation is still unknown.MethodsFOXQ1 mRNA and protein expression were analysed in a panel of CRC cell lines, and laser micro-dissected human biopsy samples by qRT-PCR, microarray GeneChip® U133 Plus 2.0 and western blots. FOXQ1 regulation was assayed by chromatin immunoprecipitation and luciferase reporter assays.ResultsFOXQ1 was robustly induced in CRC compared to other tumors, but had no predictive value with regards to grade, metastasis and survival in CRC. Prototype-based gene coexpression and gene set enrichment analysis showed a significant association between FOXQ1 and the Wnt pathway in tumors and cancer cell lines from different tissues. In vitro experiments confirmed, on a molecular level, FOXQ1 as a direct Wnt target. Analysis of known Wnt targets identified FOXQ1 as the most suitable marker for canonical Wnt activation across a wide panel of cell lines derived from different tissues.ConclusionsOur data show that FOXQ1 is one of the most over-expressed genes in CRC and a direct target of the canonical Wnt pathway. It is a potential new marker for detection of early CRC and Wnt activation in tumors of different origins.

Circulating tumour cells escape from EpCAM-based detection due to epithelial-to-mesenchymal transition

BackgroundCirculating tumour cells (CTCs) have shown prognostic relevance in metastatic breast, prostate, colon and pancreatic cancer. For further development of CTCs as a biomarker, we compared the performance of different protocols for CTC detection in murine breast cancer xenograft models (MDA-MB-231, MDA-MB-468 and KPL-4). Blood samples were taken from tumour bearing animals (20 to 200 mm2) and analysed for CTCs using 1. an epithelial marker based enrichment method (AdnaTest), 2. an antibody independent technique, targeting human gene transcripts (qualitative PCR), and 3. an antibody-independent approach, targeting human DNA-sequences (quantitative PCR). Further, gene expression changes associated with epithelial-to-mesenchymal transition (EMT) were determined with an EMT-specific PCR assay.MethodsWe used the commercially available Adna Test, RT-PCR on human housekeeping genes and a PCR on AluJ sequences to detect CTCs in xenografts models. Phenotypic changes in CTCs were tested with the commercially available “Human Epithelial to Mesenchymal Transition RT-Profiler PCR Array”.ResultsAlthough the AdnaTest detects as few as 1 tumour cell in 1 ml of mouse blood spiking experiments, no CTCs were detectable with this approach in vivo despite visible metastasis formation. The presence of CTCs could, however, be demonstrated by PCR targeting human transcripts or DNA-sequences - without epithelial pre-enrichment. The failure of CTC detection by the AdnaTest resulted from downregulation of EpCAM, whereas mesenchymal markers like Twist and EGFR were upregulated on CTCs. Such a change in the expression profile during metastatic spread of tumour cells has already been reported and was linked to a biological program termed epithelial-mesenchymal transition (EMT).ConclusionsThe use of EpCAM-based enrichment techniques leads to the failure to detect CTC populations that have undergone EMT. Our findings may explain clinical results where low CTC numbers have been reported even in patients with late metastatic cancers. These results are a starting point for the identification of new markers for detection or capture of CTCs, including the mesenchymal-like subpopulations.

Peptides in Low Molecular Weight Fraction of Serum Associated with Hepatocellular Carcinoma

The incidence of hepatocellular carcinoma (HCC) in the United States is increasing and the increase is projected to continue for several decades. The overall survival of HCC patients is poor and treatments are not effective in part because most of the diagnoses come at a late stage. The development of new markers for detection of HCC would significantly improve patient prognosis. This paper describes identification of candidate markers previously reported in our serologic study of an Egyptian population by quantitative comparison of matrix assisted laser desorption ionization time of flight (MALDI-TOF) mass spectra. To identify these marker candidates, we performed LC-MS/MS sequencing that identified nine native peptides associated with HCC, including two reported previously. Four truncations of N terminus of complement C3f and a fibrinopeptide increased in control sera; two complement C4α peptides, a zyxin peptide, and a coagulation factor XIII peptide increased in cancer patient sera. We have also identified increased biliverdin diglucuronide in the sera of cancer patients. These peptides could potentially serve as markers of HCC following additional validation studies; however, association of similar peptides with other diseases and cancers dictates a very cautious approach.

Peptides in low molecular weight fraction of serum associated with hepatocellular carcinoma.

The incidence of hepatocellular carcinoma (HCC) in the United States is increasing and the increase is projected to continue for several decades. The overall survival of HCC patients is poor and treatments are not effective in part because most of the diagnoses come at a late stage. The development of new markers for detection of HCC would significantly improve patient prognosis. This paper describes identification of candidate markers previously reported in our serologic study of an Egyptian population by quantitative comparison of matrix assisted laser desorption ionization time of flight (MALDI-TOF) mass spectra. To identify these marker candidates, we performed LC-MS/MS sequencing that identified nine native peptides associated with HCC, including two reported previously. Four truncations of N terminus of complement C3f and a fibrinopeptide increased in control sera; two complement C4α peptides, a zyxin peptide, and a coagulation factor XIII peptide increased in cancer patient sera. We have also identified increased biliverdin diglucuronide in the sera of cancer patients. These peptides could potentially serve as markers of HCC following additional validation studies; however, association of similar peptides with other diseases and cancers dictates a very cautious approach.

Fatty acid‐binding protein as marker for renal injury

The assessment of biomarkers to detect renal injury has been studied before, but the sensitivity and specificity of urinary and serum profiles of these markers still need to be improved. One of the promising new markers for detection of renal injury is the family of 15 kDa cytoplasmic fatty acid‐binding proteins (FABPs). Remarkably, however, the application of FABP as marker for renal injury due to ischaemia, toxic heavy metals or in end‐stage renal failure has only recently been investigated. The present status of two types of FABP in the kidney (heart‐type (H‐FABP), located in the distal tubular cells, and liver‐type (L‐FABP), which is located in the proximal tubular cells), is reviewed for sensitive detection of renal injury. This review is based on an overview of the literature on clinical diagnostics applying plasma and urinary H‐FABP as well as L‐FABP levels in renal toxicity, kidney transplantation or as clinical parameter in end‐stage renal failure. H‐FABP has been shown to be a proper marker for detection of ischaemic injury in tissue perfusates of non‐heart‐beating donor kidneys. Urinary L‐FABP has been evaluated recently more explicitly and shows significant elevations in progressive end‐stage renal failure as well as after ischaemic injury due to renal transplantation or cardiac bypass surgery. H‐ and L‐FABP have been shown to be useful markers for rapid detection and monitoring of renal injury. Further study of the diagnostic and prognostic use of these FABP types will require further commercialization of automated and rapid assays for proper clinical application.

UDP‐N‐acetyl‐D‐galactosamine:polypeptide N‐acetylgalactosaminyltransferase 6 (ppGalNAc‐T6) mRNA as a potential new marker for detection of bone marrow‐disseminated breast cancer cells

The evaluation of disseminated epithelial tumor cells in patients with early stages of breast cancer has generated considerable interest because of its potential association with poor clinical outcome. Considering that O-glycosylation pathways are frequently altered in breast cancer, we performed this work to evaluate the potential usefulness of UDP-N-acetyl-D-galactosamine:polypeptide N-acetylgalactosaminyltransferases (ppGalNAc-Ts) (a family of glycosyltransferases which catalyze the first key step of mucin-type O-glycosylation) to detect disseminated cells in bone marrow samples from patients with operable breast cancer. Using RT-PCR assays, we studied the gene expression of 9 enzymes (ppGalNAc-T1-T9). Among the ppGalNAc-Ts expressed by breast tumors (-T1, -T2, -T3, -T6 and -T7), the best specificity (negative results on all PBMN cell samples from healthy donors) was shown for ppGalNAc-T6. Thus, we selected this enzyme as a target gene for further evaluation. ppGalNAc-T6 mRNA was found in 22/25 (88%) breast cancer samples, in all 3 human breast cancer cell lines evaluated (MCF-7, ZR75-1 and T47D), in 1/30 (3%) PBMN cells and 0/19 bone marrow samples obtained from patients without cancer. Using this method, 22/61 (36%) patients with breast cancer, who underwent curative surgery, showed positive ppGalNAc-T6 mRNA in bone marrow aspirates obtained prior to surgery, including 11/34 patients with stage-I or -II, without histopathological lymph node involvement. In a preliminary follow-up evaluation, 19/61 patients experienced recurrence of the disease. ppGalNAc-T6 was positive in 11/19 (57.9%) of these patients. Interestingly, in the group of patients without lymph node involvement, disease recurrence was observed in 54.5% of patients who showed ppGalNAc-T6 mRNA-positive bone marrow aspirates and only in 4.3% of patients when ppGalNAc-T6 was negative (p = 0.014). These results indicate that ppGalNAc-T6 mRNA could be a specific marker applicable to the molecular diagnosis of breast cancer cells dissemination.

Cytokeratin 19 mRNA: A new marker for detection of metastases in lymph nodes of gynecological cancer patients

Cytokeratin 19 mRNA: A new marker for detection of metastases in lymph nodes of gynecological cancer patients

Fetal cells in the peripheral blood of pregnant women express thymidine kinase: a new marker for detection

Fetal cells occur in maternal blood in a substantial proportion of normal pregnancies. Several different approaches have been used to detect and enrich these cells for non-invasive prenatal diagnosis. However, before these fetal cells can routinely be used for prenatal diagnosis, perfectly reproducible procedures for detection and enrichment need to be established. We found that these fetal cells express high intracellular levels of the DNA precursor pathway enzyme thymidine kinase. Since normal adult peripheral blood cells do not exhibit any thymidine kinase activity, this enzyme is a potent new marker to detect and enrich fetal cells from maternal blood. We further describe the first successful application of a cytofluorometric thymidine kinase assay to detect fetal cells in the maternal circulation by virtue of their high thymidine kinase activity. © 1997 Federation of European Biochemical Societies.

Urinary level of L-fucose: A new marker for detection of liver damage by L-fucose tolerance test

Urinary level of L-fucose: A new marker for detection of liver damage by L-fucose tolerance test

Alpha fetoprotein in maternal serum: A new marker for detection of fetal distress and intrauterine death

Increase in circulating maternal alpha fetoprotein (AFP) is associated with intrauterine fetal death. The AFP levels in 13 women whose fetuses died in utero were from 280 to 9,000 ng. per milliliter. Nine of them (70 per cent) were above 530 ng. per milliliter, which was the upper normal level in 65 uncomplicated pregnancies. In high-risk pregnancies, the AFP test correctly predicted 60 per cent of the cases with fetal distress and 92 per cent of the cases with normal fetoplacental function. If maternal AFP levels were above 800 ng. per milliliter, fetal distress or intrauterine fetal death occurred in 85 per cent of the cases, and intrauterine fetal death occurred in all 6 cases where the maternal AFP concentration was higher than 1,075 ng. per milliliter. The increase in maternal AFP took place before the fetal death. Clinically established fetal distress was correctly predicted by the AFP test in 2 women whose estriol excretion was normal. The results suggest that the AFP test contributes to the biochemical detection of fetal distress and intrauterine death.