Value of PHI and PHID in the detection of intermediate- and high-risk prostate cancer.

BACKGROUND AND AIMS: PSA testing practice results in a large number of unnecessary prostate biopsies and the overdiagnosis of clinically insignificant prostate cancer (PCa). The aim of our study was to evaluate the value of PHI and PHID for the detection of PCa. MATERIALS AND METHODS: We measured tPSA, fPSA and p2PSA in 455 patients scheduled for biopsy, including 243 patients with PCa. D'Amico criteria were used to classify these patients in three groups related to risk of progression. Intermediate- and high-risk PCa were considered as aggressive PCa. RESULTS: The best area under the curve (AUC) value obtained in the detection of aggressive PCa was achieved for PHI and PHID (0.766 and 0.760, respectively). We found a relationship of the performance of by these tests with the calculated prostate volume or the estimated prostate size by digital rectal exam, obtaining the higher AUC in patients with a small prostate. Thus, the AUC for PHI was 0,843 for patients with small calculated prostate volume and 0,817 for patients with small estimated prostate size. CONCLUSIONS: Our results underline that PHI and PHID outperforms the efficacy obtained with tPSA and %fPSA. Substantial differences in their value in relation to prostate volume were found.

Identification of Potential miRNAs Biomarkers for High-Grade Prostate Cancer by Integrated Bioinformatics Analysis.

The increasing number of datasets available in the GEO database offers a new approach to identify new miRNAs related to PCa. The aim of our study was to suggest a miRNA signature for the detection of high-grade PCa (Gleason score ≥ 7) using bioinformatics tools. Three mRNA datasets (GSE26022, GSE30521, GSE46602) were selected to identify the differentially expressed genes (DEGs) in high-grade PCa. Furthermore, two miRNA datasets (GSE45604, GSE46738) were analyzed to select the differentially expressed miRNAs (DEMs). Functional and pathway enrichment analysis was performed using DAVID and a protein-protein interaction network (PPI) was constructed through STRING. Besides, miRNAs which regulate hub genes were predicted using microRNA.org . A total of 973 DEGs were identified after the analyses of the mRNA datasets, enriched in key mechanisms underlying PCa development. Furthermore, we identified 10 hub genes (EGFR, VEGFA, IGF1, PIK3R1, CD44, ITGB4, ANXA1, BCL2, LPAR3, LPAR1). The most significant KEGG Pathway was PI3K-Akt signaling pathway, involved in cell proliferation and survival. Moreover, we identified 30 common miRNAs between significant DEMs and the predicted hub gene regulators. Twelve of these miRNAs (miR-1, -365, -132, -195, -133a, -133b, -200c, -339, -222, -21, -221, -708) regulate two or more hub genes identified in our study. We suggested a signature including these 12 miRNAs for high-grade PCa detection. These miRNAs have been associated with aggressive PCa, poor survival and resistance to treatment in the last years.

Novel Biomarkers for Prostate Cancer Detection and Prognosis.

Prostate cancer (PCa) remains as one of the most controversial issues in health care because of the dilemmas related to screening using Prostate Specific Antigen (PSA). A high number of false positive biopsies and an elevated rate of overdiagnosis are the main problems associated with PSA. New PCa biomarkers have been recently proposed to increase the predictive value of PSA. The published results showed that PCA3 score, Prostate Health Index and 4Kscore can reduce the number of unnecessary biopsies, outperforming better than PSA and the percentage of free PSA. Furthermore, 4Kscore provides with high accuracy an individual risk for high-grade PCa. High values of PHI are also associated with tumor aggressiveness. In contrast, the relationship of PCA3 score with aggressiveness remains controversial, with studies showing opposite conclusions. Finally, the development of molecular biology has opened the study of genes, among them TMPRSS2:ERG fusion gene and miRNAs, in PCa detection and prognosis.

Emerging biomarkers in the diagnosis of prostate cancer.

Prostate cancer (PCa) is the second most common cancer in men worldwide. A large proportion of PCa are latent, never destined to progress or affect the patients' life. It is of utmost importance to identify which PCa are destined to progress and which would benefit from an early radical treatment. Prostate-specific antigen (PSA) remains the most used test to detect PCa. Its limited specificity and an elevated rate of overdiagnosis are the main problems associated with PSA testing. New PCa biomarkers have been proposed to improve the accuracy of PSA in the management of early PCa. Commercially available biomarkers such as PCA3 score, Prostate Health Index (PHI), and the four-kallikrein panel are used with the purpose of reducing the number of unnecessary biopsies and providing information related to the aggressiveness of the tumor. The relationship with PCa aggressiveness seems to be confirmed by PHI and the four-kallikrein panel, but not by the PCA3 score. In this review, we also summarize new promising biomarkers, such as PSA glycoforms, TMPRSS2:ERG fusion gene, microRNAs, circulating tumor cells, androgen receptor variants, and PTEN gene. All these emerging biomarkers could change the management of early PCa, offering more accurate results than PSA. Nonetheless, large prospective studies comparing these new biomarkers among them are required to know their real value in PCa detection and prognosis.

Concordance between direct and indirect measurements of free 25-OH vitamin D.

At present, data comparing the quantification methods for measurement of free vitamin D (direct assay [direct 25-OHDF] and estimated by calculation [calculated 25-OHDF]), are scarce. The aim of this study was to analyse the concordance between these two methods of 25-OHDF analysis (direct vs. calculated). METHODS: Serum values of total 25-OHD (25-OHDT), vitamin D binding protein (DBP) (by R&D Systems ELISA), calculated 25-OHDF (by DBP, albumin and 25-OHDT) and direct 25-OHDF (by DIAsource ELISA) were analysed in 173 healthy women (aged 35-45years). Assessment of concordance was evaluated by the Bland-Altman plot and the total deviation index (TDI). RESULTS: The mean values of calculated and direct 25-OHDF in these subjects were 5.27±2.5 and 3.83±1.01pg/mL, respectively. We found significantly lower values of 25-OHDF on comparing subjects with and without vitamin D deficiency, independently of the method used. The total deviation index evaluated by the Bland-Altman plot showed low concordance for both measurements. Only low 25-OHDF levels were concordant. CONCLUSIONS: This study shows that the concordance between these two methods of 25-OHDF analysis is low and has a concentration dependent bias. Further studies are necessary to clarify the reference values and the indications for 25-OHDF measurement.

miRNAs as novel biomarkers in the management of prostate cancer.

microRNAs (miRNAs) are small non-coding RNAs that control gene expression posttranscriptionally and are part of the giant non codifying genoma. Cumulating data suggest that miRNAs are promising potential biomarkers for many diseases, including cancer. Prostate cancer (PCa) detection is currently based in the serum prostate-specific antigen biomarker and digital rectal examination. However, these methods are limited by a low predictive value and the adverse consequences associated with overdiagnosis and overtreatment. New biomarkers that could be used for PCa detection and prognosis are still needed. Recent studies have demonstrated that aberrant expressions of microRNAs are associated with the underlying mechanisms of PCa. This review attempts to extensively summarize the current knowledge of miRNA expression patterns, as well as their targets and involvement in PCa pathogenesis. We focused our review in the value of circulating and urine miRNAs as biomarkers in PCa patients, highlighting the existing discrepancies between different studies, probably associated with the important methodological issues related to their quantitation and normalization. The majority of studies have been performed in serum or plasma, but urine obtained after prostate massage appears as a new way to explore the usefulness of miRNAs. Large screening studies to select a miRNA profile have been completed, but bioinformatics tools appear as a new approach to select miRNAs that are relevant in PCa development. Promising preliminary results were published concerning miR-141, miR-375 and miR-21, but larger and prospective studies using standardized methodology are necessary to define the value of miRNAs in the detection and prognosis of PCa.

Exosomal and Non-Exosomal Urinary miRNAs in Prostate Cancer Detection and Prognosis.

BACKGROUND: MicroRNAs (miRNAs) are non-coding small RNAs, involved in post-transcriptional regulation of many target genes. METHODS: Five miRNAs that have been consistently found deregulated in PCa (miR-21, miR-141, miR-214, miR-375, and let-7c) were analyzed in urinary pellets from 60 prostate cancer (PCa) patients and 10 healthy subjects by qRT-PCR. Besides, urinary exosomes were isolated by differential centrifugation and analyzed for those miRNAs. RESULTS: Significant upregulation of miR-21, miR-141, and miR-375 was found comparing PCa patients with healthy subjects in urinary pellets, while miR-214 was found significantly downregulated. Regarding urinary exosomes, miR-21 and miR-375 were also significantly upregulated in PCa but no differences were found for miR-141. Significant differences were found for let-7c in PCa in urinary exosomes while no differences were observed in urinary pellets. A panel combining miR-21 and miR-375 is suggested as the best combination to distinguish PCa patients and healthy subjects, with an AUC of 0.872. Furthermore, the association of miRNAs with clinicopathological characteristics was investigated. MiR-141 resulted significantly correlated with Gleason score in urinary pellets and let-7c with clinical stage in urinary exosomes. Additionally, miR-21, miR-141, and miR-214 were found significantly deregulated in intermediate/high-risk PCa versus low-risk/healthy subjects in urinary pellets. Significant differences between both groups were found in urinary exosomes for miR-21, miR-375, and let-7c. CONCLUSIONS: These findings suggest that the analysis of miRNAs-especially miRNA-21 and miR-375- in urine could be useful as biomarkers in PCa. Prostate 77: 573-583, 2017. © 2016 Wiley Periodicals, Inc.

Prostate Cancer Detection and Prognosis: From Prostate Specific Antigen (PSA) to Exosomal Biomarkers.

Prostate specific antigen (PSA) remains the most used biomarker in the management of early prostate cancer (PCa), in spite of the problems related to false positive results and overdiagnosis. New biomarkers have been proposed in recent years with the aim of increasing specificity and distinguishing aggressive from non-aggressive PCa. The emerging role of the prostate health index and the 4Kscore is reviewed in this article. Both are blood-based tests related to the aggressiveness of the tumor, which provide the risk of suffering PCa and avoiding negative biopsies. Furthermore, the use of urine has emerged as a non-invasive way to identify new biomarkers in recent years, including the PCA3 and TMPRSS2:ERG fusion gene. Available results about the PCA3 score showed its usefulness to decide the repetition of biopsy in patients with a previous negative result, although its relationship with the aggressiveness of the tumor is controversial. More recently, aberrant microRNA expression in PCa has been reported by different authors. Preliminary results suggest the utility of circulating and urinary microRNAs in the detection and prognosis of PCa. Although several of these new biomarkers have been recommended by different guidelines, large prospective and comparative studies are necessary to establish their value in PCa detection and prognosis.

Automated measurement of 25-OH Vitamin D on the LUMIPULSE® G1200: analytical verification and method comparison.

BACKGROUND: Because of its potential value in several pathologies, clinical interest in 25-hydroxy Vitamin D (25OH-D) is increasing. However, the standardisation of assays remains a significant problem. Our aim was to evaluate the performance of the novel Lumipulse G 25-OH Vitamin D assay (Fujirebio), comparing results with the Liaison (Diasorin) method. METHODS: Analytical verification of the Lumipulse G 25-OH Vitamin D assay was performed. Both methods were compared using sera from 226 patients, including 111 patients with chronic renal failure (39 on haemodialysis) and 115 patients without renal failure. In addition, clinical concordance between assays was assessed. RESULTS: For Lumipulse G 25-OH Vitamin D assay, the limit of detection was 0.3 ng/mL, and the limit of quantification was 2.5 ng/mL with a 9.7% of coefficient of variation. Intra-and inter-assay coefficients of variation were <2.3 and <1.8% (25.4-50.0 ng/mL), respectively. Dilution linearity was in the range of 4.5-144.5 ng/mL. Method comparison resulted in a mean difference of -6.5% (95% CI from -8.8 to -4.1) for all samples between Liaison and Lumipulse G. Clinical concordance assessed by Kappa Index was 0.66. CONCLUSIONS: Lumipulse G 25-OH Vitamin D showed a good clinical concordance with the Liaison assay, although overall results measured in Lumipulse were higher by an average of 6.5%.

Emerging biomarkers in the detection and prognosis of prostate cancer.

The prostate-specific antigen (PSA) is currently the most used tumor marker in the early detection of the prostate cancer (PCa), despite its low specificity and low negative predictive value. New biomarkers, including urine prostate cancer gene 3 (PCA3) score, Prostate Health Index (PHI), and the four-kallikrein panel, have been investigated during recent years especially with the aim of detecting aggressive PCa. Results suggest the ability of these biomarkers to improve the specificity of PSA in the detection of PCa, although there are not enough results directly comparing these biomarkers to know their complementarity. The relationship with PCa aggressiveness seems to be confirmed for PHI and for the four-kallikrein panel, but not for PCA3 score. However, available results suggest that emerging biomarkers may be useful as part of a multivariable approach for the screening and prognosis of PCa. Nevertheless, larger prospective studies comparing these biomarkers are necessary to evaluate definitely their value in the management of early PCa.

The influence of prostate volume in prostate health index performance in patients with total PSA lower than 10 µg/L.

Prostate health index (phi), a measure calculated as p2PSA/fPSA × âˆštPSA, has shown valuable results in the detection of prostate cancer (PCa), improving the prediction of the aggressiveness of the tumor. The aim of our study was to test whether prostate volume influences phi performance using univariate and multivariate models. 220 patients with PSA<10 µg/L (102 with negative biopsy and 118 with PCa) were included in the study. Serum concentrations of tPSA, fPSA and p2PSA were measured on Access2 analyzer. The higher accuracy was found for phi, obtaining an AUC of 0.748. Bigger AUCs were obtained for phi, %p2PSA, %fPSA and tPSA in patients with small prostate volume (≤35 cc); meanwhile, the lowest AUCs were found in patients with large prostate volume (>50 cc). Including phi and %p2PSA in a multivariable analysis based on patient age, prostate volume, tPSA, and %fPSA accuracy increased from 0.762 to 0.802 (logistic regression model) or 0.815 (artificial neural network). Accuracy excluding prostate volume in these models was 0.762 and 0.775, respectively. The inclusion of phi and %p2PSA in a multivariate model identifies better men with PCa. Prostate volume remains a key factor in the interpretation of biomarkers used to detect PCa.

Real-time PCR PCA3 assay is a useful test measured in urine to improve prostate cancer detection.

BACKGROUND: The usefulness of PCA3 in the management of early prostate cancer (PCa) is on debate. The aim of this study was to evaluate PCA3 in the detection of PCa and its relationship with tumor aggressiveness. METHODS AND PATIENTS: PCA3 score was measured by real time PCR in urinary sediments of 122 patients who underwent prostate biopsy for PSA >4µg/L. RESULTS: Analysis of ROC curves showed an area under the curve (AUC) of 0.804 for PCA3 score, while the AUCs were 0.587 and 0.697 for PSA and % free PSA, respectively. The probability of positive biopsy increased in relation to PCA3 score, with variations from 37% to 96% for patients with PCA3 score lower than 0.90 or higher than 1.04, respectively. We chose the cut-off value of 0.90, corresponding to a sensitivity of 92.5%, for which we obtained a specificity of 41.5%. No significant differences in PCA3 score were found in relation to Gleason score or clinical stage. CONCLUSIONS: The results show a high probability of PCa in patients with an elevated PCA3 score, although we did not find any relationship with aggressiveness of the tumor. PCA3 score can help to select more precisely which patients will need to perform a biopsy of the prostate.

Clinical utility of %p2PSA and prostate health index in the detection of prostate cancer.

BACKGROUND: %p2PSA and prostate health index (phi) has shown valuable results in the detection of prostate cancer (PCa), improving the prediction of the aggressiveness of the tumor. The goal of the present study was to evaluate %p2PSA and phi in the detection of PCa, estimating their relationship with the aggressiveness of PCa. METHODS: A total of 354 patients with positive or negative prostatic biopsy were included. Prospectively, 150 were enrolled and 204 were enrolled retrospectively proceeding from our serum bank. RESULTS: The best performance was observed for %p2PSA and phi, obtaining an AUC of 0.723 and 0.732, respectively. The highest specificity at sensitivity around 90% was obtained for phi (27.4%). Using the cut-off of 31.94 for phi, a reduction of 19% biopsies could be obtained, while 17 PCa would have been missed, including only four patients with a Gleason score ≥7. Similarly, using a cut-off of 1.21 for %p2PSA, a reduction of 12.7% biopsies could be obtained, while 16 PCa would have been missed, including only four patients with a Gleason score ≥7. Moreover, among patients with PCa, phi (median: 69.75 vs. 48.04) and %p2PSA (median: 2.60 vs. 1.98) values are significantly higher (p<0.0001) in patients with a biopsy Gleason score ≥7. CONCLUSIONS: Our results confirm previous evaluations, showing similar AUCs and results in sensitivity and specificity to other studies.%p2PSA and phi raise the accuracy in the detection of prostate cancer, reducing the number of unnecessary biopsies and improving the prediction of the aggressiveness of the tumor.

ProPSA, un nuevo biomarcador para la detección y el manejo del cáncer de próstata / ProPSA, a new biomarker for the detection and management of prostate cancer

La descripción de las nuevas isoformas del PSA libre, como el proPSA, que se asocia a la presencia de cáncer de próstata, ha ampliado las herramientas disponibles para la detección de este tumor. Tras la comercialización de un ensayo para medir [---2] proPSA, una de las fracciones del proPSA, disponemos de datos que avalan el empleo del porcentaje de [---2] proPSA en relación con el PSA libre (%[---2] proPSA) y del prostate health index (phi) que valora conjuntamente [---2] proPSA, PSA libre y PSA total. Los resultados disponibles indican que estos tests permiten reducir el número de biopsias negativas cuando se comparan con el porcentaje del PSA libre. Por otro lado, tanto el %[---2] proPSA como el phi se relacionan con tumores particularmente agresivos, por lo que podrían ser tests útiles para seleccionar qué pacientes podrían beneficiarse de una vigilancia activa y qué pacientes deben ser sometidos a un tratamiento curativo (AU)

The description of new PSA isoforms associated with prostate cancer, such as proPSA, has expanded the available tools for the detection of this tumor. Since the marketing of an assay for the measurement of [---2] proPSA, one of the fractions of proPSA, we have positive data on the use of the percentage of [---2] proPSA in relation to free PSA (%[---2] proPSA) and the prostate health index (phi), which measures [---2] proPSA, total PSA and free PSA together. The available results suggest that the use of these tests would lead to a reduction in the number of negative biopsies compared with the percentage of free PSA. On other hand, both %[---2] proPSA and phi are related to particularly aggressive tumors, so they could be useful to select patients for active surveillance, and to decide which patients must be treated (AU)

PCA3 in the detection and management of early prostate cancer.

Although widely used, the value of prostate-specific antigen (PSA) in the detection of prostate cancer is controversial. The percentage of free PSA increases the specificity of PSA, but results are not enough. Prostate cancer gene 3 (PCA3) has been proposed as an option that may complement these markers in the detection and management of early prostate cancer. Our aim has been to review the value of PCA3 as tumor marker. The available results suggest that PCA3 is particularly useful to select in which patients the biopsy should be repeated when the first biopsy was negative. However, some points should be specified with further studies, including the most appropriate PCA3 cutoff level and the significance of a high PCA3 score in patients with negative biopsy. False-negative results are also a conflictive point in the use of PCA3, because prostate cancer, including aggressive tumors, may be present in patients with a low PCA3 score. Probably, a proper interpretation of this test requires its management together with other tests, through multivariate models for the detection of prostate cancer. On the other hand, several studies showed the relation between PCA3 score and Gleason score, and also the utility of PCA3 to select patients for active surveillance. To summarize, the available studies show the utility of PCA3 in the detection and management of early prostate cancer, although some aspects referred to its use need to be retested after further studies to confirm the actual value and the limitations of this test.

HE4 a novel tumour marker for ovarian cancer: comparison with CA 125 and ROMA algorithm in patients with gynaecological diseases.

The aim of this study is to evaluate a new tumour marker, HE4, in comparison with CA 125 and the Risk of Ovarian Malignancy Algorithm (ROMA) in healthy women and in patients with benign and malignant gynaecological diseases. CA 125 and HE4 serum levels were determined in 66 healthy women, 285 patients with benign gynaecological diseases (68 endometriosis, 56 myomas, 137 ovarian cysts and 24 with other diseases), 33 patients with non-active gynaecological cancer and 143 with active gynaecological cancer (111 ovarian cancers). CA 125 and HE4 cut-offs were 35 U/mL and 150 pmol/L, respectively. ROMA algorithm cut-off was 13.1 and 27.7 for premenopausal or postmenopausal women, respectively. HE4, CA 125 and ROMA results were abnormal in 1.5%, 13.6% and 25.8% of healthy women and in 1.1%, 30.2% and 12.3% of patients with benign diseases, respectively. Among patients with cancer, HE4 (in contrast to CA 125) had significantly higher concentrations in ovarian cancer than in other malignancies (p < 0.001). Tumour marker sensitivity in ovarian cancer was 79.3% for HE4, 82.9% for CA 125 and 90.1% for ROMA. Both tumour markers, HE4 and CA 125 were related to tumour stage and histological type, with the lowest concentrations in mucinous tumours. A significantly higher area under the ROC curve was obtained with ROMA and HE4 than with CA 125 in the differential diagnosis of benign gynaecological diseases versus malignant ovarian cancer (0.952, 0.936 and 0.853, respectively). Data from our population indicate that ROMA algorithm might be further improved if it is used only in patients with normal HE4 and abnormal CA 125 serum levels (cancer risk for this profile is 44.4%). ROMA algorithm in HE4 positive had a similar sensitivity and only increases the specificity by 3.2% compared to HE4 alone.

Day-to-day variation of late-night salivary cortisol in healthy voluntaries.

OBJECTIVES: The study examines the components of biological variation of nocturnal salivary cortisol in healthy subjects. DESIGN AND METHODS: Eight repetitive measurements were performed in seven subjects during a 25-day time period (study A), and then, for comparison, two salivary specimens were taken during two consecutive days from 20 subjects (study B). Salivary cortisol was measured with the Salimetrics HS-Cortisol assay. RESULTS: Mean salivary cortisol (1.27 nmol/L), analytical variation (CVa=15.4%), within-subject variation (CVi=34.1%), between-subject variation (CVg=35.3%), index of individuality (II=1.06) and reference change value (RCV=104%) were obtained for study A. Similar results were obtained from the set of samples of study B. CONCLUSION: The study results show a medium degree of individuality for salivary cortisol. Both conventional reference values and comparison of serial results may be equally used for clinical interpretation. A change greater of 104% between two successive measurements should be considered significant.