EPCAM Germ Line Deletions as Causes of Lynch Syndrome in Spanish Patients

The standard genetic test for Lynch syndrome (LS) frequently reveals an absence of pathogenic mutations in DNA mismatch repair genes known to be associated with LS. It was recently shown that germ line deletions in the last exons of EPCAM are involved in the etiology of LS. The aim of this study was to evaluate the prevalence of EPCAM deletions in a Spanish population and the clinical implications of deletion. Probands from 501 families suspected of having LS were enrolled in the study. Twenty-five cases with MSH2 loss were identified: 10 had mutations of MSH2, five had mutations of MSH6, and 10 did not show MSH2/MSH6 mutations. These 25 cases were analyzed for EPCAM deletions using multiplex ligation-dependent probe amplification, and deletions were mapped using long-range PCR analysis. One subject with no MSH2/MSH6 mutations had a large deletion in the EPCAM locus that extended for 8.7 kb and included exons 8 and 9. The tumor exhibited MSH2 promoter hypermethylation. EPCAM deletion analysis followed by MSH2 methylation testing of the tumor is a fast low-cost procedure that can be used to identify mutations that cause LS. We propose that this procedure be incorporated into clinical genetic analysis strategies and present a decision-support flow diagram for the diagnosis of LS.Lynch syndrome (LS) is an autosomal dominant inherited cancer syndrome characterized by early-onset cancers of the colorectum and endometrium and tumors of the stomach, pancreas, small intestine, ovary, bladder, and bile duct.¹ In the Spanish population, about 2.5% of colorectal cancers are associated with LS.² The carcinogenetic etiology of this syndrome involves a DNA mismatch repair (MMR) inactivation caused by a germ line mutation of an MMR gene (MLH1, MSH2, MSH6, or PMS2) followed by somatic inactivation of the second allele.¹ As a consequence of MMR inactivation, these tumors exhibit microsatellite instability (MSI) and loss of expression of the mutated MMR gene.¹ It was recently shown that germ line deletions involving the last exon of the non-MMR gene, EPCAM (OMIM#185535), may silence its neighboring gene, MSH2 (OMIM#609309), which is located 17 kb downstream of EPCAM, via promoter hypermethylation. This epigenetic inactivation seems to be effective only in tissues in which EPCAM is expressed.^3,4 The EPCAM gene codes for the epithelial cell adhesion molecule also known as CD326, which is expressed in all normal epithelial cells and in carcinoma tumors.⁵ Thus, deletions of the last exon of EPCAM constitute a distinct class mutation associated with LS.Currently, the standard genetic test for LS (point mutation and large-rearrangement analysis of MLH1, MSH2, MSH6, and PMS2) frequently fails to detect a pathogenic mutation. For this reason, we evaluated the association between EPCAM deletions and LS in a Spanish population and its clinical implications.     

Candidemia in pediatric patients with congenital heart disease

Candidemia is an important problem in pediatrics. In our hospital, highest candidemia rates were documented among children with congenital heart disease (CHD). A series was conducted to describe the clinical and mortality features of candidemia in these patients. Fifty-two cases (1988-2000) included very young infants (median age, 2 months) who received long-term antibiotic treatment (median, 20.5 days). Candida parapsilosis predominated (54%). Endovascular infections occurred in 11.5%. In-hospital mortality was 39% and related mortality 14%. Maintenance of catheter (odds ratio [OR], 6.0; 95% confidence interval [CI], 1.0-37.2; P = .05) and severity of patients as measured with the Pediatric Risk Score of Mortality I (OR, 1.1, 95% CI, 1.0-1.3; P = .05) were independently associated with mortality. In summary, candidemia in children with CHD is diagnosed to very young infants with prolonged antibiotic therapy. Mortality is high but, in most cases, is not related to candidemia. Optimal management may include exclusion of endocarditis, early antifungal treatment, and catheter removal.     

Adherence to the Dutch Breast Cancer Guidelines for Surveillance in Breast Cancer Survivors: Real-World Data from a Pooled Multicenter Analysis

BackgroundRegular follow-up after treatment for breast cancer is crucial to detect potential recurrences and second contralateral breast cancer in an early stage. However, information about follow-up patterns in the Netherlands is scarce.Patients and MethodsDetails concerning diagnostic procedures and policlinic visits in the first 5 years following a breast cancer diagnosis were gathered between 2009 and 2019 for 9916 patients from 4 large Dutch hospitals. This information was used to analyze the adherence of breast cancer surveillance to guidelines in the Netherlands. Multivariable logistic regression was used to relate the average number of a patient’s imaging procedures to their demographics, tumor–treatment characteristics, and individual locoregional recurrence risk (LRR), estimated by a risk-prediction tool, called INFLUENCE.ResultsThe average number of policlinic contacts per patient decreased from 4.4 in the first to 2.0 in the fifth follow-up year. In each of the 5 follow-up years, the share of patients without imaging procedures was relatively high, ranging between 31.4% and 33.6%. Observed guidelines deviations were highly significant (P < .001). A higher age, lower UICC stage, and having undergone radio- or chemotherapy were significantly associated with a higher chance of receiving an imaging procedure. The estimated average LRR-risk was 3.5% in patients without any follow-up imaging compared with 2.3% in patients with the recommended number of 5 imagings.ConclusionCompared to guidelines, more policlinic visits were made, although at inadequate intervals, and fewer imaging procedures were performed. The frequency of imaging procedures did not correlate with the patients’ individual risk profiles for LRR.     

Carbonaceous Materials Porosity Investigation in a Wet State by Low-Field NMR Relaxometry

The porosity of differently wetted carbonaceous material with disordered mesoporosity was investigated using low-field ¹H NMR relaxometry. Spin–spin relaxation (relaxation time T₂) was measured using the CPMG pulse sequence. We present a non-linear optimization method for the conversion of relaxation curves to the distribution of relaxation times by using non-specialized software. Our procedure consists of searching for the number of components, relaxation times, and their amplitudes, related to different types of hydrogen nuclei in the sample wetted with different amounts of water (different water-to-carbon ratio). We found that a maximum of five components with different relaxation times was sufficient to describe the observed relaxation. The individual components were attributed to a tightly bounded surface water layer (T₂ up to 2 ms), water in small pores especially supermicropores (2 < T₂ < 7 ms), mesopores (7 < T₂ < 20 ms), water in large cavities between particles (20–1500 ms), and bulk water surrounding the materials (T₂ > 1500 ms). To recalculate the distribution of relaxation times to the pore size distribution, we calculated the surface relaxivity based on the results provided by additional characterization techniques, such as thermoporometry (TPM) and N₂/−196 °C physisorption.