Endocan - a potential diagnostic marker for early onset sepsis in neonates.

INTRODUCTION: Neonatal early onset sepsis assessment is based on the history of pregnancy and delivery and nonspecific clinical signs. None of the biomarkers currently in use for clinical practice has adequate prognostic value, so it is not possible to clearly distinguish neonates with culture-proven sepsis from those with only risk factors or clinical suspicion. Endocan is an endothelial mediator involved in the inflammatory response that is present in low concentrations in the serum of healthy subjects, and in much higher concentrations in patients with SIRS and septic shock. The purpose of this study is to evaluate the utility of serum endocan serum levels as a biomarker for the diagnosis of neonatal early onset sepsis (EOS). METHODOLOGY: Serum endocan concentration was measured in newborns with clinical suspicion of EOS admitted to the Neonatal Intensive Care Unit on day 1, 3 and 7. RESULTS: Serum endocan levels were significantly increased in septic compared to non-septic neonates in the early stages of sepsis (2.43 ± 0.95 vs. 1.77 ± 0.57, p = 0.004), continued to rise up to 72 hours from onset and then decreased by the seventh day under treatment. CONCLUSIONS: These results suggest a potential role for endocan as an early marker for diagnosis and follow-up in neonatal EOS. Studies on a larger number of cases are needed in order to establish the practical utility of this molecule as a diagnostic tool for clinical practice.

Comparison of next generation sequencing, SNaPshot assay and real-time polymerase chain reaction for lung adenocarcinoma EGFR mutation assessment.

BACKGROUND: The epidermal growth factor receptor (EGFR) mutation status assessment has become increasingly important given the significant impact of tyrosine kinase inhibitors in lung cancer management. Our aim was to compare real life operational characteristics for three EGFR mutation assays - two targeted approaches and a next generation sequencing (NGS) technique. METHODS: EGFR mutation status was assessed for lung adenocarcinoma samples (formalin fixed- paraffin embedded samples) using qPCR, SNaPshot and NGS (Ion Torrent™) techniques. RESULTS: A total of 15 high clinical significance mutations were identified by at least one technique from the total of 64 samples. All mutations were identified by the TaqMan qPCR technique while SNaPshot in conjunction with fragment analysis identified 11 EGFR mutations. The NGS approach followed by an automatic analysis using the default calling parameters identified 10 mutations from the SNaPshot/qPCR panel and other three insertions, five point mutations and 58 silent variants; manual data review identified all 15 high significance mutations. CONCLUSIONS: Performance was similar for high tumor content samples but careful data analysis and post hoc variant calling filter alterations were necessary in order to obtain robust results from low tumor content samples by NGS. NGS is able to generate a comprehensive mutational profile albeit at a higher cost and workload. Result interpretation should take into account not only general run parameters such as mean read depth but also relative coverage and read distribution; currently there is an acute need to define firm recommendations/standards concerning NGS data interpretation and quality control.

Thoracic oncology HERMES: European curriculum recommendations for training in thoracic oncology.

Thoracic oncology HERMES: European curriculum recommendations for training in thoracic oncology http://ow.ly/mdqT300NHqO.

Defect in recruiting effector memory CD8+ T-cells in malignant pleural effusions compared to normal pleural fluid.

BACKGROUND: Malignant pleural effusions (MPE) are a common and fatal complication in cancers including lung or breast cancers, or malignant pleural mesothelioma (MPM). MPE animal models and immunotherapy trials in MPM patients previously suggested defects of the cellular immunity in MPE. However only few observational studies of the immune response were done in MPM patients, using questionable control groups (transudate…). METHODS: We compared T cell populations evaluated by flow cytometry from blood and pleural effusion of untreated patients with MPM (n = 58), pleural metastasis of adenocarcinoma (n = 30) or with benign pleural lesions associated with asbestos exposure (n = 23). Blood and pleural fluid were also obtained from healthy subjects, providing normal values for T cell populations. RESULTS: Blood CD4+ or CD8+ T cells percentages were similar in all groups of patients or healthy subjects. Whereas pleural fluid from healthy controls contained mainly CD8+ T cells, benign or malignant pleural effusions included mainly CD4+ T cells. Effector memory T cells were the main T cell subpopulation in pleural fluid from healthy subjects. In contrast, there was a striking and selective recruitment of central memory CD4+ T cells in MPE, but not of effector cells CD8+ T cells or NK cells in the pleural fluid as one would expect in order to obtain an efficient immune response. CONCLUSIONS: Comparing for the first time MPE to pleural fluid from healthy subjects, we found a local defect in recruiting effector CD8+ T cells, which may be involved in the escape of tumor cells from immune response. Further studies are needed to characterize which subtypes of effector CD8+ T cells are involved, opening prospects for cell therapy in MPE and MPM.

[New diagnostic markers for malignant pleural mesothelioma]. / Nouveaux marqueurs diagnostiques du mésothéliome pleural malin.

The increasing number of malignant pleural mesothelioma (MPM) is a serious public health problem. The prognosis of this tumor is poor and most of the patients are diagnosed late in the disease's course when curative treatment is no more an option. It is therefore necessary to diagnose earlier MPM in these patients in order to obtain a potential significant improvement in survival. Some serum markers have been previously proposed for MPM diagnosis but none had sufficient sensitivity and specificity for being use in routine. Recently, soluble mesothelin and osteopontin have been proposed as diagnostic markers for mesothelioma. The authors reviewed recent data concerning the utility of these two molecules in the diagnosis and the treatment of MPM. Mesothelin seems to be a specific marker for the epithelioid subtype of mesothelioma. Despite a good sensitivity, osteopontin has a low specificity for mesothelioma diagnosis. However, there is not enough data yet to propose guidelines on the use of these markers in a day to day practice.

Loss of Endocan tumorigenic properties after alternative splicing of exon 2.

BACKGROUND: Endocan was originally described as a dermatan sulfate proteoglycan found freely circulating in the blood. Endocan expression confers tumorigenic properties to epithelial cell lines or accelerate the growth of already tumorigenic cells. This molecule is the product of a single gene composed of 3 exons. Previous data showed that endocan mRNA is subject to alternative splicing with possible generation of two protein products. In the present study we identified, and functionally characterized, the alternative spliced product of the endocan gene: the exon 2-deleted endocan, called endocanDelta2. METHODS: Stable, endocanDelta2-overexpressing cell lines were generated to investigate the biological activities of this new alternatively spliced product of endocan gene. Tumorigenesis was studied by inoculating endocan and endocanDelta2 expressing cell lines subcutaneously in SCID mice. Biochemical properties of endocan and endocanDelta2 were studied after production of recombinant proteins in various cell lines of human and murine origin. RESULTS: Our results showed that the exon 2 deletion impairs synthesis of the glycan chain, known to be involved in the pro-tumoral effect of endocan. EndocanDelta2 did not promote tumor formation by 293 cells implanted in the skin of severe combined immunodeficient (SCID) mice. CONCLUSION: Our results emphasize the key role of the polypeptide sequence encoded by the exon 2 of endocan gene in tumorigenesis, and suggest that this sequence could be a target for future therapies against cancer.

[Comparative evaluation between MB/BacT system and Löwenstein-Jensen solid culture media for mycobacterial isolation]. / Evaluare comparativa a sistemului MB/BacT cu mediul Löwenstein-Jensen pentru izolarea micobacteriilor.

Tuberculosis is a public health issue in both developed and developing countries. Success of the treatment depend on the identification of patients with positive sputum smears, rapid confirmation of diagnosis in patients with negative microscopy and identification of mycobacterial strains with altered drug susceptibility. Data from the literature show that liquid culture media have a higher sensitivity for isolation mycobacteria than solid culture media as Löwenstein-Jensen. In our series inoculation on liquid media resulted in retrieval of a significant higher number of mycobacterial strains than on solid media (435 vs 250). The time needed to obtain a positive culture was also lower for liquid media (15.89 +/- 9 days, mean +/- standard deviation) than for solid media (27.77 +/- 10.13 days), p < 0.001. These differences were seen in both smear negative and smear positive cases. Culture in liquid media isolated more strains with altered drug susceptibility but this difference was not statistically significant.

Utility of osteopontin and serum mesothelin in malignant pleural mesothelioma diagnosis and prognosis assessment.

PURPOSE: Malignant mesothelioma is a highly aggressive tumor and is often diagnosed too late for a curative treatment. We compared diagnostic and prognostic values of mesothelin and osteopontin in 172 patients suspected of malignant pleural mesothelioma (MPM) and in a control group of 112 asymptomatic asbestos-exposed subjects. EXPERIMENTAL DESIGN: Osteopontin and mesothelin were assayed with commercial ELISA kits in a series of 43 patients with pleural metastases of various carcinomas, 33 patients with benign pleural lesions associated with asbestos exposure, 96 patients with MPMs, and 112 asbestos-exposed healthy subjects. Results were correlated with patient's diagnosis and survival. RESULTS: Serum osteopontin level was higher in MPM patients compared with healthy asbestos-exposed subjects and had a good capability to distinguish between these two populations. However, osteopontin was unable to distinguish between MPM and pleural metastatic carcinoma or benign pleural lesions associated with asbestos exposure. Neither plasma nor pleural fluid osteopontin were more powerful in this respect. Serum mesothelin had a good ability for diagnosing MPM but was unable to identify patients with nonepithelioid mesothelioma subtypes. Survival analysis identified tumor histologic subtype along with serum osteopontin and serum mesothelin as independent prognostic factors in mesothelioma patients. CONCLUSIONS: Osteopontin has a lower diagnostic accuracy than mesothelin in patients suspected of MPM. Insufficient specificity limits osteopontin utility as diagnostic marker. Both molecules have a potential value as prognostic markers.

Endocan expression and relationship with survival in human non-small cell lung cancer.

PURPOSE: We evaluated the expression of endocan, a soluble lung- and kidney-selective endothelial cell-specific dermatan sulfate proteoglycan, in non-small cell lung tumors compared with normal lung and studied the significance of high levels of circulating endocan in patients with non-small cell lung cancer. MATERIAL AND METHODS: Endocan and vascular endothelial growth factor mRNA expression were evaluated by semiquantitative PCR in tumoral and nontumoral lung tissue samples from a first series of 24 patients submitted to curative surgery. Relationships between survival, time to tumor progression, and serum levels of endocan were evaluated in a second series of 30 previously untreated patients addressed for staging. RESULTS: In non-small cell lung cancers, endocan mRNA was overexpressed compared with control lung. Immunohistochemistry shows that endocan was expressed only by tumor endothelium in all cases, especially in the periphery of the tumors, with no differences between adenocarcinoma and squamous cell carcinoma. Endocan and vascular endothelial growth factor mRNA expression was positively correlated in lung tumors. Serum endocan levels, as well as tumor, node, and metastasis status, were correlated with both survival and time to tumor progression. However, endocan serum level was not an independent prognostic factor due to its correlation with the presence of metastasis. CONCLUSION: Endocan is overexpressed in non-small cell lung tumors compared with healthy lung and probably represents a response of tumoral endothelium to proangiogenic growth factor stimulation. Circulating levels of endocan might reflect tumor angiogenic stimulation and present prognostic significance.