Current Biomarkers - Volume 6, Issue 1, 2016

Nitric Oxide (NO) is an important, rapid assay biomarker of many known diseases, including asthma, chronic obstructive pulmonary disease (COPD) as well as other respiratory infections, septic shock, Parkinson's diseases and Alzheimer's dementia. Therefore, the detection and measurement of NO in a physiological fluid, either in gas or in liquid phase, are scientifically and clinically significant. The objective of this research is to develop a single-use, disposable cost effective in vitro NO biosensor which can measure NO of ppb (parts per billion) level in gas phase and subμM (sub micro mole) level in liquid phase. This biosensor is a thin film platinum based electrochemical sensor. Ni (II) tetrakis (3-methoxy-4-hydroxyphenlyl) porphyrin is mixed with diethylene glycol and Nafion forming a NO selective membrane-electrolyte. Reactive sputtering, laser ablation, thick film printing technologies are used to fabricate the biosensor on a roll-to-roll cost effective processing. Differential pulse voltammetry (DPV) is employed as the measurement technique. The results of this NO biosensor show that this biosensor can measure NO down to 5ppb in gas phase and 0.00242μM of NO in PBS solution (liquid phase). Fabrication and test procedures of this NO biosensor are described in this presentation. In conclusion, this development is scientifically successful and is described as sensor for nitric oxide detection WO2015143197 (2015) by World Intellectual Property Organization on September 24, 2015.

Background: Thyroid cancers are frequent in the endocrine system especially differentiated thyroid carcinoma and 95% of all thyroid cancer cases are differentiated neoplasmas. They have a favorable prognosis; however although, a small group present recurrence in 10-15% of patients following the standard treatment, surgery and radioiodine. Medullary thyroid carcinoma which is a neoplasm of the parafollicular C cells of the thyroid is responsible for a high number of thyroid cancer related deaths. Objective: Nowadays, the therapeutic options for advanced differentiated neoplasms, metastatic disease or anaplastic carcinoma are scarce. Recently, multitargeted kinase inhibitors are considered as new treatments for differentiated thyroid neoplasms and have been introduced in their management. The American Thyroid Association recommendations stated in the guideline of 2015 that tyrosine kinase inhibitors should be used in the treatment of patients with refractory differentiated thyroid carcinoma and in those cases that present progression. Methods: In this review, the purpose is to analyze the thyroid cancer tumorigeniesis and the action of the different targeted agents over these pathways of activation and the results in those cases of thyroid cancer without other possible treatments. Results: Multikinase inhibitors, the most widely studied, are able in thyroid cancer to interfere in the proliferation, invasion, and neoangiogenesis of neoplastic cells in thyroid cancer. Recently, the US Food and Drug Administration and the European Medicine Agency approved sorafenib and lenvatinib, which in Europe received an orphan designation by European Medicine Agency for follicular thyroid cancer. Some other controlled trials with tyrosine kinase inhibitors, as vandetanib and carbozantinib, have been finished and are now the two approved drugs in advanced medullary thyroid cancers. Conclusion: Thus, drug targeting represents a challenging approach with promising potential to circumvent some problems associated with many toxic effects of antineoplastic drugs.

Gastrointestinal cancers remain a significant health problem worldwide. Resistance to treatment in a subset of patients is a major problem and personalized treatment is expected to improve prognosis in these individuals. Assessment of biomarkers, which are generally genes, proteins, phosphoproteins and/or microRNAs (miRNAs), and their levels in tumor or blood, leads to a view of the molecular characteristics (signature) of a given patient’s tumor and thus provides information that will enable personalized treatment. miRNAs are short noncoding RNAs that regulate gene expression at the post-transcriptional level. Deregulation of miRNA expression has been demonstrated in various cancers, including gastrointestinal (GI) cancers. The high stability of miRNAs in the blood circulation offers enormous advantage for prognostic applications as blood samples can be easily procured noninvasively. Circulating miRNAs have been demonstrated to be valuable instruments in prognosis of GI cancers. Additionally, serial collection of blood samples is possible for monitoring the progress of cancer, something that cannot be done reasonably with tumor tissue. We review previous reports that used circulating miRNAs as biomarkers in the prognosis of GI cancers, and also highlight the fascinating possibility of using the gathered information to identify new therapeutic targets.

Background: CD133 is a widely used cancer stem cell marker for many cancers. Scientometric analysis is a reliable and practical method that enables us to get more information on CD133. To our knowledge, there is no bibliometric study on CD133. So, an overview of the general trend of CD133 is important for scientific researchers. Methods: In this study, data extraction was performed using the semantic search engine ‘GoPubMed’. Papers were collected from the PubMed database since 2000 to 2015. Major topics of CD133 were summarized by using Carrot. Results: Top subject area were cancers (lung cancer, colon cancer, hepatocellular, glioma, and laryngeal cancer), genes (CD117, CD90, ALDH1, Oct4, and c-Myc), and signaling pathways (wnt and mTOR). Highly-productive authors, and countries, and journals were ranked. Then, a term map was constructed and clusters by using VOSviewer mapping software. Conclusion: Our work can provide a systematic view to the researchers without prior working experience in CD133 field. CD133 is a widely used cancer stem cell marker for many cancers. Scientometric analysis is a reliable and practical method that enables us to get more information on CD133. To our knowledge, there is no bibliometric study on CD133. So, an overview of the general trend of CD133 is important for scientific researchers. In this study, data extraction was performed using the semantic search engine ‘GoPubMed’. Papers were collected from the PubMed database since 2000 to 2015. Major topics of CD133 were summarized by using Carrot. Top subject areas were cancers (lung cancer, colon cancer, hepatocellular, glioma, and laryngeal cancer), genes (CD117, CD90, ALDH1, Oct4, and c-Myc), and signaling pathways (wnt and mTOR). Highly-productive authors, and countries, and journals were ranked. Then, a term map was constructed and clusters by using VOSviewer mapping software. Our work can provide a systematic view to the researchers without prior working experience in CD133 field.

Background: Carbonic anhydrase is a metalloenzyme extensively present in nature. In animals, it is involved in several functions such as respiration, pH homeostasis, salt transport, metabolic reactions, calcification, bone resorption. In recent years, the CA wide distribution, relevance in several physiological processes, involvement in several pathological conditions, as well as sensitivity to chemical pollutants has driven the research on CA based biomarkers. Objective: The review is addressed to analyze the potential applicability of CA as biomarker in a variety of fields, from human health to environmental sciences. Results: The main studies in this field are analyzed and discussed in the present review exploring several areas of interest, from clinic to environmental monitoring. The review also details interesting research patents, which have yielded in the last years in this research field. Conclusion: In conclusion, the review highlights that CA is becoming a promising biomarker in several areas of interest and outlines that the research on CA based biomarkers have opened new perspective for translation of advances in basic science into innovative applications and patents.

Background: The problem of diabetes and associated treatments had mentioned herbal plants in several studies. The cardioactive Terminalia arjuna has also been reported for antidiabetic principles yet no scientific evidence is available. Hence, an attempt has been made by the authors to find the probable mechanism and biomarker involved for α-amylase enzyme inhibition Objective: The extraction of bark of Terminalia arjuna using room temperature extraction (RTE) and soxhlet extraction (SE), estimation of total percentage yield, the percentage yield of marker compounds arjunic acid and arjunolic acid present and in vitro evaluation of α-amylase inhibitory activity of the extracts has been investigated. Method: The solvent system used for HPTLC was toluene: acetic acid: ethyl acetate (5: 5: 0.5) and the corresponding spots to the marker compounds were quantitatively estimated via standard plot of both the marker compounds. The molecular docking was performed in dry lab to evaluate the effect of arjunolic and arjunic acid on the α-amylase enzyme protein. Scanning Electron Microscopy was done to study the effect of extraction methods on crude and exhausted drug. The in vitro α-amylase inhibitory activity of the two extracts was performed spectrophotometrically taking acarbose as standard. Results: The total percentage yield was 2.8% and 6.7%, while the percentage yield of the marker compounds was 0.63% and 0.98% of arjunic acid whereas 0.87% and 1.12% of arjunolic acid in RTE and SE respectively. The molecular docking suggested that the arjunolic acid (-47.703Kcal/mol) exhibited better results of inhibition compared with the free energy values of arjunic acid (-30.836Kcal/mol) and standard acarbose (-43.439Kcal/mol). Scanning Electron Microscopy showed that the SE exhibited more rupturing of cell walls than RTE and leads to better extraction of phytoconstituents. The in vitro α-amylase inhibitory activity of the two extracts and the results of IC50 value for the two extracts showed better inhibition viz. 76.12 + 0.40μ g/mL and 65.61 + 0.47μg/mL for RTE and SE respectively as compared to standard acarbose 83.18 + 0.38μg/mL. Conclusion: Hence, there exists correlation between the higher anti-diabetic potential of SE of Terminalia arjuna plant, as compared to RTE on the basis of α-amylase inhibition as demonstrated by molecular docking and wet lab results.

Background: In acute coronary syndrome (ACS), nitric oxide is generated in excess during ischemic attack due to marked activation of the inducible nitric oxide synthase (iNOS) enzyme by different cytokines. Consequently, serious damaging effects result. Objective: To explore the potential role of iNOS mRNA expression as a marker of acute ischemia in ACS patients and as a predictor of the attack outcome. Method: Forty-eight ACS patients and 24 healthy volunteers were enrolled in the study. Cardiac markers were assessed, including creatine kinase (CK), CK-MB and troponin T. RNA was extracted from peripheral blood and iNOS mRNA was quantified by reverse transcription and real-time PCR (RT-PCR). Results: iNOS mRNA relative expression was significantly elevated in ACS patients compared to healthy subjects (p < 0.001) and was lower in patients with good outcome than those with bad outcome (p = 0.029). ROC curve discriminated between ACS patients and controls (AUROC curve: 1; p < 0.001) with 100% sensitivity and specificity at a cutoff value of 38.5. iNOS expression in ACS patients with normal troponin T (66.7%) and those with elevated levels were comparable (p = 0.24). In addition, iNOS expression showed no correlation with CK and CK-MB levels. Conclusion: The estimation of iNOS mRNA expression is an applicable, sensitive and specific assay for ACS diagnosis. Low mRNA levels correlate with good outcome of the acute ischemic attack. However, further study on larger sample size can validate these findings.

Setting: We investigated the usefulness of bone markers, respectively Procollagen Type I Aminoterminal Propeptide (PINP) and Beta-Crosslaps (β-CTx) for diagnosis, treatment and monitoring of charges with histologically confirmed breast neoplasm. The intention of this project was the demonstration of the impact of the osseous fluctuation markers PINP on the one hand and β-CTx on the other hand for the verification of bone metastases in female patients suffering from breast cancer. PINP is a marker for bone generation and specific indicator of type I collagen deposition. β-CTx are delivered as disruption artefacts of collagen type I and therefore able to be detected in serum and urine. Objectives: The main objective of the following thesis was the evaluation of the significance of bone markers β-CTx and PINP regarding their advantage for matutinal recognition of osseous expansion and pathological skeletal reorganization in pre- and postmenopausal breast cancer patients. Patients, Materials and Methods: Peripheral blood samples of each 80 patients with known breast neoplasm were amassed and the bone markers PINP and β-CTx quantified. Therefore we used a specific immunoassay “ECLIA” and the analysis approach ELECSYS 2010 and COBAS e by Roche Diagnostics (Mannheim, Germany). Our groups of patients were divided with regard to menopausal and bone metastases status. Results: By contrast with the group devoid of bone involvement with a median of 73.61ng/ml there are significant higher serum concentrations of PINP in the female collective with scintigraphically verified skeletal metastases with a median of 125.75ng/ml. In any case, both populations revealed data above the provided cutoff values (premenopausal → 27.8ng/ml and postmenopausal → 37.1ng/ml).For β-CTx the study depicted similar results: β-CTx provided a specificity and sensitivity of 100.0%. An increased β-CTx level was connected significantly with osseous metastases in our study collectives (median: 67.0ng/ml) in contrast to the bone metastases-negative population (median: 0.905ng/ml). Moreover there were also significant differences concerning the menopausal degree of development of patients (p < 0.001). Conclusion: Summing up the markers of bone turnover PINP and β-CTx could be considered as useful and helpful diagnostic tools for detecting bone involvement in patients with known breast carcinoma. As indicators for bone metabolism, formation and resorption they are up to externalize a significant discrimination between breast cancer patients with and without bone infestation. In addition they seem to be beneficial to ascertain these sick persons with early dysfunctions in skeletal metabolism and help to initiate premature attendance. It is an important discussion point whether these bone markers are capable to supplant conventional procedures for the registration of bone metastases like the radiological ones.