Monitoring of graft function is essential during the first months after liver transplantation (LT), but current liver function tests (LFTs) lack the specificity and sensitivity to ensure an efficient ...diagnosis of acute rejection (AR). Recently, donor‐derived cell‐free DNA (ddcfDNA) has emerged as a noninvasive biomarker to assess graft integrity. This study evaluated the feasibility of measuring the ddcfDNA through short tandem repeat (STR) analysis by quantitative fluorescent‐polymerase chain reaction (QF‐PCR) and to assess the role of the concentration and fragment size of total cfDNA as AR biomarkers. The total concentration and fragment size of cfDNA and the ddcfDNA percentage were monitored in plasma of 20 patients without rejection and 7 patients with T‐cell–mediated AR during the first 3 months after LT. The median ddcfDNA percentage was 3‐fold higher before AR diagnosis (34.8%; P < 0.001) and moderately higher at AR confirmatory diagnosis (23.8%; P = 0.049) compared with that of nonrejector patients (10.6%), showing a better performance (area under the curve = 84.6%) than conventional LFTs to predict the risk of rejection within the first 2 weeks following LT. The fraction of 100‐250‐bp cfDNA fragments was higher at AR diagnosis compared with that of nonrejector patients (68.0% versus 57.9%, P = 0.02). STR amplification by QF‐PCR may be an alternative strategy for rapid ddcfDNA quantification, which is easily implementable in clinical laboratories. The results of this pilot study indicate that ddcfDNA increases very early, even 1‐2 weeks before the diagnosis of AR, and so it could be useful as a prognostic biomarker in improving patient risk stratification.
https://www.wileyhealthlearning.com/#/online-courses/d1f49d22-fe8c-44ee-b5c9-5a26cd1abeda
Tumor markers are a heterogeneous group of substances released by cancer cells into bloodstream, but also expressed by healthy tissues. Thus, very small concentrations can be present in plasma and ...serum from healthy subjects. Cancer patients tend to show increased levels correlating with tumor bulk, but false positive results could be present in patients with benign conditions. The correct interpretation of TM results could be challenging and many factors should be considered, from pre-analytical conditions to patient concomitant diseases. In this line, the Clinical Chemistry and Laboratory Medicine journal has made important contributions though several publications promoting the adequate use of TM and therefore improving patient safety. TM measurement offers valuable information for cancer patient management in different clinical contexts, such as helping diagnosis, estimating prognosis, facilitating early detection of relapse and monitoring therapy response. Our review analyzes the clinical usefulness of tumor markers applied in most frequent epithelial tumors, based on recent evidence and guidelines.
RNA helicase DHX15 plays a significant role in vasculature development and lung metastasis in vertebrates. In addition, several studies have demonstrated the overexpression of DHX15 in the context of ...hepatocellular carcinoma. Therefore, we hypothesized that this helicase may play a significant role in liver regeneration, physiology, and pathology.
gene deficiency was generated by CRISPR/Cas9 in zebrafish and by TALEN-RNA in mice. AUM Antisense-Oligonucleotides were used to silence
in wild-type mice. The hepatocellular carcinoma tumor induction model was generated by subcutaneous injection of Hepa 1-6 cells. Homozygous
gene deficiency was lethal in zebrafish and mouse embryos.
gene deficiency impaired liver organogenesis in zebrafish embryos and liver regeneration after partial hepatectomy in mice. Also, heterozygous mice presented decreased number and size of liver metastasis after Hepa 1-6 cells injection compared to wild-type mice.
gene silencing with AUM Antisense-Oligonucleotides in wild-type mice resulted in 80% reduced expression in the liver and a significant reduction in other major organs. In addition,
gene silencing significantly hindered primary tumor growth in the hepatocellular carcinoma experimental model. Regarding the potential use of DHX15 as a diagnostic marker for liver disease, patients with hepatocellular carcinoma showed increased levels of DHX15 in blood samples compared with subjects without hepatic affectation. In conclusion, Dhx15 is a key regulator of liver physiology and organogenesis, is increased in the blood of cirrhotic and hepatocellular carcinoma patients, and plays a key role in controlling hepatocellular carcinoma tumor growth and expansion in experimental models.
•Prostatic ischemia and cellular damage can occur during cardiogenic shock.•Increased PSA serum levels during cardiogenic shock might be associated with prostatic cellular damage.•Cardiogenic shock ...is a cause of PSA false positive results.•High PSA concentrations in patients with cardiogenic shock cannot be considered as a marker of PCa.
Prostate-specific antigen (PSA) is the tumor marker most widely used in conjunction with digital rectal examination (DRE) for the early detection of prostate cancer (PCa). Due to its limitations, especially the high rate of false positive (FP) results, PSA screening of transplant candidates is a controversial issue. Moreover, obtaining a FP result in the PCa screening of heart transplant candidates may lead to potentially harmful effects. Although most of the factors that may cause PSA FP results are well known, FP results related to cardiogenic shock, a common indication for heart transplant, are less known. We studied retrospectively four patients who suffered cardiogenic shock during their hospital stay and became heart transplant candidates. Their PSA serum levels were very high suggesting the presence of PCa. Our findings have shown that elevated PSA serum levels in these patients were not related to PCa and they might be associated with cardiogenic shock. This clinical case study adds evidences to the fact that cardiogenic shock is an important cause of PSA FP results, therefore it cannot be used as a reliable marker of PCa in this clinical condition and positive results should be properly interpreted.
This study is the first verification of the novel iPTH Siemens ADVIA Centaur® Intact Parathyroid Hormone (iPTHm) chemiluminescence immunoassay based on monoclonal antibodies. We also compared the ...iPTH results obtained using this assay with the previous ADVIA Centaur® Parathyroid Hormone assay (iPTHp) based on polyclonal antibodies.
The analytical performance study of iPTHm assay included LoD, LoQ, intra- and inter-assay reproducibility, and linearity. A comparison study was performed on 369 routine plasma samples. The results were analyzed independently for patients with normal and abnormal GFR, as well as patients on hemodialysis. In addition, clinical concordance between assays was assessed. Finally, we studied PTH stability of plasma samples at 4°C.
For the iPTHm assay LoD and LoQ were 0.03pmol/L and 0.10pmol/L, respectively. Intra- and inter-assay CV were between 2.3% and 6.2%. Linearity was correct in the range from 3.82 to 203.08pmol/L. Correlation studies showed a good correlation (r=0.99) between iPTHm and iPTHp, with bias of −2.55% (IC −3.48% to −1.62%) in the range from 0.32 to 117.07pmol/L. Clinical concordance, assessed by Kappa Index, was 0.874. The stability study showed that differences compared to basal iPTH concentration did not exceed 20% in any of the samples analyzed.
The iPTHm assay demonstrated acceptable performance and a very good clinical concordance with iPTHp assay, currently used in our laboratory. Thus, the novel iPTHm assay can replace the previous iPTHp assay, since results provided by both assays are very similar. In our study, the stability of iPTH is not affected by storage up to 14days.
•A verification of the novel iPTH Siemens ADVIA Centaur® Intact Parathyroid Hormone (iPTHm) has been performed.•Accurate analytical performance, and excellent agreement and clinical concordance with iPTHp assay have been demonstrated.•Novel iPTHm assay can replace the previous iPTHp assay.•The stability of iPTH (plasma, 4°C) is not affected by storage up to 14days.
Chronic liver disease and related complications, cirrhosis and hepatocellular carcinoma, are associated with high mortality. Curative treatments, partial hepatectomy or liver transplantation, have ...limited applicability in patients with cirrhosis due to the poor liver regenerative capacity. Thus, we need to find new diagnostic and therapeutic alternatives, to block the disease progression and to improve the survival of patients. In this context, preclinical studies have demonstrated the key role of the protein kinase B (Akt) in liver dysfunction, but the status of Akt and its targets in patients with chronic hepatopathy remains unknown. Aims: To determine the activation status of the Akt pathway and their association with liver functionality in cirrhotic patients.
This retrospective study includes liver tissue samples from 36 hepatectomized patients with (n=27) and without (n=9) cirrhosis. Multiplex analysis of proteins involved in the Akt/mTOR pathway was performed using a Luminex panel and Western blot. Conventional liver function tests were determined in serum before resection surgery.
Akt and forkhead box protein O1 (FoxO1) are overexpressed in the liver of cirrhotic patients: (2.1 vs. 1.0 densitometric relative units (DRU); p<0.01, and 9.5 vs
4.4 DRU; p<0.01, respectively). FoxO1 showed the best correlation with markers of liver injury (aspartate aminotransferase (ASAT): r=0.51, p<0.05; alanine aminotransferase (ALAT): r=0.49, p<0.05), and was the only enzyme in the Akt pathway identified as an independent predictor of ASAT and ALAT levels.
The intrahepatic expression of FoxO1 could have clinical utility as a potential prognostic marker for patients with advanced liver disease.
High circulating levels of 2-hydroxyglutarate (2HG) have been reported in patients with determinate isocitrate dehydrogenase (IDH) mutated tumors. Recent studies indicate that in malignancies such as ...acute myeloid leukemia (AML), measurements of 2HG in serum provide useful diagnostic and prognostic information and improve patient selection and monitoring of IDH-targeted treatments. In the current study, we validated a sensitive and specific gas chromatography mass spectrometry (GC–MS) method specifically intended to quantify serum levels of 2HG in routine clinical laboratories. Extraction was liquid-liquid with ethyl acetate, and derivatization was reduced to 3 min of microwave irradiation. The analytical method was linear over a wide dynamic range, presenting acceptable intraday and day-to-day precision and accuracy. The limit of quantification was 10 ng/mL, process efficiency ranged between 38% and 49%, and recovery of added 2HG was 99–105%. 2HG was found to be stable in serum for up to 48 h at both 4 °C and at ambient temperature, and after three freeze-thaw cycles. Microwave derivatizated extracts in the autosampler were found to be stable for up to 120 h. In summary, the present method is useful for quantification of 2HG serum levels in patients with IDH mutated malignancies in clinical laboratories.
•Serum 2-hydroxyglutarate (2HG) is a clinically useful biomarker in some malignancies.•We report a GC–MS method for the determination of 2HG levels in human serum.•Derivatization was reduced to a 3 minute microwave irradiation.•The method is useful for serum 2HG measurements in clinical laboratories.
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,
fusion gene, microRNAs, circulating tumor cells, androgen receptor variants, and
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.
Resumen
Objetivos
La enfermedad hepática crónica y sus complicaciones, la cirrosis y el carcinoma hepatocelular, presentan una elevada mortalidad. Los tratamientos curativos, como la hepatectomía ...parcial o el trasplante hepático, tienen una aplicación limitada en pacientes con cirrosis, por su escasa capacidad de regeneración hepática. Son necesarias otras alternativas diagnósticas y terapéuticas para prevenir la progresión de la enfermedad y mejorar la supervivencia. Diversos estudios preclínicos demuestran el importante papel de la proteína quinasa B(Akt) en la disfunción hepática, aunque aún se desconoce el estado de Akt y sus dianas en las patologías hepáticas crónicas. El principal objetivo es determinar el estado de activación de la vía Akt y su relación con la función hepática en pacientes cirróticos.
Métodos
Estudio retrospectivo con muestras de tejido hepático de 36 pacientes hepatectomizados con (n=27) y sin (n=9) cirrosis. Se realizó un análisis Multiplex de las proteínas de la vía Akt/mTOR empleando un panel Luminex y
Western blot
. Previamente a la resección, se realizaron las pruebas habituales de función hepática en suero.
Resultados
Akt y la proteína FoxO1 están sobreexpresadas en el hígado cirrótico: 1.0 unidades densitométricas relativas (UDR); p<0,01, y 9,5 vs
.
4,4 DRU; p<0,01, respectivamente). FoxO1 mostró una fuerte correlación con los marcadores de daño hepático (aspartato aminotransferasa (ASAT): r=0,51, p<0,05; alanina aminotransferasa (ALAT): r=0,49, p<0,05), y fue la única enzima de la vía Akt identificada como predictor independiente de los niveles de ASAT y ALAT.
Conclusiones
La expresión intrahepática de FoxO1 podría tener utilidad clínica como marcador pronóstico en la enfermedad hepática avanzada.
Introduction: 6-Monoacetylmorphine (6-MAM) is a specific metabolite of heroin. Thus, the presence of 6-MAM in urine is a definitive indication of heroin intake. The possibility of having an ...immunoassay procedure to measure 6-MAM would be a diagnosis tool to discriminate, among opiates-positive, those patients who have consumed heroin and those who have not.
Methods: EMIT
®
II Plus 6-Acetylmorphine Assay was used to measure 6-MAM in urine. The positive opiate screening results were confirmed at the Toxicology laboratory of our hospital by GC-MS.
Results: This study includes 63 urine samples from subjects admitted to emergency department with suspicion of opiate consumption. Specificity was evaluated in the two groups of samples studied. In the first group all samples which resulted negative by opiate immunoassay (n = 33) were negative for 6-MAM immunoassay test. Thus, the specificity obtained for 6-MAM immunoassay in this group was 100%. Regarding the second specificity study, performed in positive samples by opiate immunoassay which were negative to 6 MAM by GC-MS, the specificity decreased down to 75%. In the study of sensitivity all samples confirmed as positive to 6-MAM by confirmatory method (GC-MS) resulted positive by the screening method, thus sensitivity obtained was 100%.
Discussion: In this study no FN for 6-MAM was observed and therefore the new Emit
®
II Plus 6- Acetylmorphine Assay procedure has a high NPV, thus a negative result with 6-MAM immunoassay practically excludes heroine consume. The positive results to 6-MAM by immunoassay should be confirmed by a more analytically specific method, such as GCMS.
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