The distribution of extracellular matrix in the human uterus

Several lines of evidence suggest that there may be a distinction between the inner and outer myometrium. Magnetic resonance imaging has shown that the uterus of women of reproductive age consists of three layers, in addition, transvaginal ultrasound has revealed that peristaltic waves, during the course of the normal menstrual cycle, emanate only from the inner myometrial muscle. Finally histological findings have suggested that there is a three-fold increase in the nuclear density of the inner compared to the outer myometrium, hi addition, trophoblast invasion is restricted to the inner third of the myometrium. Based on these lines of evidence it was postulated that there could be a difference in extracellular matrix between inner and outer myometrial smooth muscle. To test this hypothesis, the distribution of different laminin chains, collagen IV and elastin were examined in the human uterus, hi addition observations were made to determine whether there was tissue specificity of laminin type expression. Forty-four hysterectomy specimens were collected, from women undergoing hysterectomy for benign conditions, representing all phases of the menstrual cycle. These also included specimens from patients who had been treated with intrauterine levonorgestrel (MirenaRTM). Cryo- and paraffin embedded sections were prepared. Immunocytochemistry was carried out using monoclonal antibodies directed to the alpha2, beta1, beta2, and gamma1 laminin chains, collagen IV, elastin, CD31 and the 68kD neurofilament protein. Digital imaging, by microscopy and scanning, was undertaken and novel image analysis methods were developed to examine the myometrial distribution of extracellular matrix proteins. Elastin detection was confirmed by orcein staining. As predicted collagen IV and the gamma1 laminin chain were present in the basement membranes of the vascular smooth muscle, myometrial smooth muscle, vascular-endothelium and endometrial epithelium. (Abstract shortened by ProQuest.)

Mutation analysis of the BRCA1 and BRCA2 genes in Turkish patients with breast cancer.

Annual Meeting of the American-Society-of-Clinical-Oncology (ASCO) / Clinical Science Symposium on Predicting and Improving Adverse Outcomes in Older Adults with Cancer -- MAY 29-JUN 02, 2015 -- Chicago, ILWOS: 000358036902345Amer Soc Clin Onco

The Utility of NGS Analysis in Homologous Recombination Deficiency Tracking

Several tumor types have been efficiently treated with PARP inhibitors (PARPis), which are now approved for the treatment of ovarian, breast, prostate, and pancreatic cancers. The BRCA1/2 genes and mutations in many additional genes involved in the HR pathway may be responsible for the HRD phenomenon. The aim of the present study was to investigate the association between genomic loss of heterozygosity (gLOH) and alterations in 513 genes with targeted and immuno-oncology therapies in 406 samples using an NGS assay. In addition, the %gLOHs of 24 samples were calculated using the Affymetrix technology in order to compare the results obtained via the two methodologies. HR variations occurred in 20.93% of the malignancies, while BRCA1/2 gene alterations occurred in 5.17% of the malignancies. The %LOH was highly correlated with alterations in the BRCA1/2 genes, since 76.19% (16/21) of the BRCA1/2 positive tumors had a high %LOH value (p = 0.007). Moreover, the LOH status was highly correlated with the TP53 and KRAS statuses, but there was no association with the TMB value. Lin’s concordance correlation coefficient for the 24 samples simultaneously examined via both assays was 0.87, indicating a nearly perfect agreement. In conclusion, the addition of gLOH analysis could assist in the detection of additional patients eligible for treatment with PARPis

Clinical feasibility of NGS liquid biopsy analysis in NSCLC patients.

BackgroundAnalysis of circulating tumor nucleic acids in plasma of Non-Small Cell Lung Cancer (NSCLC) patients is the most widespread and documented form of "liquid biopsy" and provides real-time information on the molecular profile of the tumor without an invasive tissue biopsy.MethodsLiquid biopsy analysis was requested by the referral physician in 121 NSCLC patients at diagnosis and was performed using a sensitive Next Generation Sequencing assay. Additionally, a comparative analysis of NSCLC patients at relapse following EGFR Tyrosine Kinase Inhibitor (TKIs) treatment was performed in 50 patients by both the cobas and NGS platforms.ResultsAt least one mutation was identified in almost 49% of the cases by the NGS approach in NSCLC patients analyzed at diagnosis. In 36 cases with paired tissue available a high concordance of 86.11% was observed for clinically relevant mutations, with a Positive Predictive Value (PPV) of 88.89%. Furthermore, a concordance rate of 82% between cobas and the NGS approach for the EGFR sensitizing mutations (in exons 18, 19, 21) was observed in patients with acquired resistance to EGFR TKIs, while this concordance was 94% for the p.T790M mutation, with NGS being able to detect this mutation in three 3 additional patients.ConclusionsThis study indicates the feasibility of circulating tumor nucleic acids (ctNA) analysis as a tumor biopsy surrogate in clinical practice for NSCLC personalized treatment decision making. The use of new sensitive NGS techniques can reliably detect tumor-derived mutations in liquid biopsy and provide clinically relevant information both before and after targeted treatment in patients with NSCLC. Thus, it could aid physicians in treatment decision making in clinical practice