Differential associations of SLCO transporters with prostate cancer aggressiveness between African Americans and European Americans

Background: Androgen receptor signaling is crucial to prostate cancer aggressiveness. Members of the solute carrier family of the organic anion transporting peptides (SLCO) are potential regulators of androgen availability in prostate tissue. It remains unknown whether genetic variations in SLCOs contribute to the differences in prostate cancer aggressiveness in African Americans (AA) and European Americans (EA). Methods: SNPs in 11 SLCO members were selected, with addition of 139 potentially functional SNPs and 128 ancestry informative markers. A total of 1,045 SNPs were genotyped and analyzed in 993 AAs and 1,057 EAs from the North Carolina–Louisiana Prostate Cancer Project. Expression and cellular localization of SLCOs were examined using qRT-PCR, IHC, and in situ RNA hybridization in independent sets of prostate cancer cases. Results: Significant associations with prostate cancer characteristics were found for SNPs in SLCO2A1 and SLCO5A1. The associations differed by race (Pinteraction < 0.05). SNPs in SLCO2A1 were associated with reduced tumor aggressiveness and low Gleason score in AAs; whereas, SNPs in SLCO5A1 were associated with high clinical stage in EAs. In prostate tissue, SLCO2A1 and SLCO5A1 were the most expressed SLCOs at the mRNA level and were expressed predominantly in prostate endothelial and epithelial cells at the protein level, respectively. Conclusions: SLCO2A1 and SLCO5A1 play important but different roles in prostate cancer aggressiveness in AAs versus EAs. Impact: The finding calls for consideration of racial differences in biomarker studies of prostate cancer and for investigations on functions of SLCO2A1 and SLCO5A1 in prostate cancer

Thioredoxin 1 in prostate tissue is associated with gleason score, erythrocyte antioxidant enzyme activity, and dietary antioxidants

Background. Prostate cancer is the most common noncutaneous cancer and second leading cause of cancer-related mortality in men in the US. Growing evidence suggests that oxidative stress is involved in prostate cancer. Methods. In this study, thioredoxin 1 (Trx 1), an enzyme and subcellular indicator of redox status, was measured in prostate biopsy tissue from 55 men from the North Carolina-Louisiana Prostate Cancer Project. A pathologist blindly scored levels of Trx 1. The association between Trx 1 and the Gleason score, erythrocyte antioxidant enzyme activity, and dietary antioxidant intake was determined using Fisher's exact test. Results. Trx 1 levels in benign prostate tissue in men with incident prostate cancer were positively associated with the Gleason score (P=0.01) and inversely associated with dietary antioxidant intake (P=0.03). In prostate cancer tissue, Trx 1 levels were associated with erythrocyte glutathione peroxidase activity (P=0.01). No association was found for other erythrocyte enzymes. Greater Gleason score of malignant tissue corresponds to a greater difference in Trx 1 levels between malignant and benign tissue (P=0.04). Conclusion. These results suggest that the redox status of prostate tissue is associated with prostate cancer grade and both endogenous and exogenous antioxidants

Evaluation of appendicitis risk prediction models in adults with suspected appendicitis

Background Appendicitis is the most common general surgical emergency worldwide, but its diagnosis remains challenging. The aim of this study was to determine whether existing risk prediction models can reliably identify patients presenting to hospital in the UK with acute right iliac fossa (RIF) pain who are at low risk of appendicitis. Methods A systematic search was completed to identify all existing appendicitis risk prediction models. Models were validated using UK data from an international prospective cohort study that captured consecutive patients aged 16–45 years presenting to hospital with acute RIF in March to June 2017. The main outcome was best achievable model specificity (proportion of patients who did not have appendicitis correctly classified as low risk) whilst maintaining a failure rate below 5 per cent (proportion of patients identified as low risk who actually had appendicitis). Results Some 5345 patients across 154 UK hospitals were identified, of which two‐thirds (3613 of 5345, 67·6 per cent) were women. Women were more than twice as likely to undergo surgery with removal of a histologically normal appendix (272 of 964, 28·2 per cent) than men (120 of 993, 12·1 per cent) (relative risk 2·33, 95 per cent c.i. 1·92 to 2·84; P < 0·001). Of 15 validated risk prediction models, the Adult Appendicitis Score performed best (cut‐off score 8 or less, specificity 63·1 per cent, failure rate 3·7 per cent). The Appendicitis Inflammatory Response Score performed best for men (cut‐off score 2 or less, specificity 24·7 per cent, failure rate 2·4 per cent). Conclusion Women in the UK had a disproportionate risk of admission without surgical intervention and had high rates of normal appendicectomy. Risk prediction models to support shared decision‐making by identifying adults in the UK at low risk of appendicitis were identified

Metanephric adenoma: the utility of immunohistochemical and cytogenetic analyses in differential diagnosis, including solid variant papillary renal cell carcinoma and epithelial-predominant nephroblastoma

Metanephric adenoma is a benign renal neoplasm that overlaps in morphology with the solid variant of papillary renal cell carcinoma and epithelial-predominant nephroblastoma. To aid in resolving this differential diagnosis, we investigated the utility of immunohistochemical and molecular analyses in distinguishing between these entities; the first study, to our knowledge, to use a combined approach in analyzing all three tumors. We analyzed 37 tumors originally diagnosed as metanephric adenomas (2 of which we reclassified as papillary renal cell carcinomas), 13 solid variant papillary renal cell carcinomas, and 20 epithelial-predominant nephroblastomas using a combination of immunohistochemistry and fluorescence in situ hybridization (FISH) assessing for trisomy of chromosomes 7 and 17 and loss of Y. Immunohistochemical staining was performed for CK7, AMACR, WT1, and CD57. The combination of CK7-, AMACR-, WT1+, and CD57+ was considered characteristic of metanephric adenoma. Most of the tumors originally diagnosed as metanephric adenomas (31/37) showed the expected staining pattern of metanephric adenoma (CK7-, AMACR-, WT1+, and CD57+). Of the six tumors with discordant immunophenotype, two tumors were reclassified as papillary renal cell carcinoma after cytogenetic workup. It is recommended that all adult cases histologically resembling metanephric adenoma have WT1, CD57, CK7, and AMACR immunohistochemical staining performed. If the staining pattern is characteristic for metanephric adenoma (CK7-, AMACR-, WT1+, and CD57+, including membranous staining), then no other diagnostic tests are indicated. However, if there is a different immunostaining pattern, then we recommend FISH analysis

Patient-derived organoid biobank identifies epigenetic dysregulation of intestinal epithelial MHC-I as a novel mechanism in severe Crohn's Disease.

Peer reviewed: TrueAcknowledgements: The authors would like to thank Prof Gillian Griffith (Cambridge Institute for Medical Research, CIMR, Cambridge) for providing helpful guidance and access to tissue from OTI mice. We thank the University of Lausanne for sharing transgenic mice and Dr. A. Zenobi (IRB Bellinzona) who helped with the mouse shipment’s organization. We would like also to acknowledge Sophie Pritchard (Cellular Generation and Phenotyping core facility at the Wellcome Sanger Institute) and Dr. Heather Zecchini (CRUK Cambridge Institute Light Microscopy facility) for helping us with RNA scope and spinning disk microscopy image acquisition and analysis respectively. Furthermore, we thank Prof Louise Boyle (Department of Pathology, University of Cambridge) for advice and constructive discussions. The illustrative graphics of experiments were created with BioRender.com.OBJECTIVE: Epigenetic mechanisms, including DNA methylation (DNAm), have been proposed to play a key role in Crohn's disease (CD) pathogenesis. However, the specific cell types and pathways affected as well as their potential impact on disease phenotype and outcome remain unknown. We set out to investigate the role of intestinal epithelial DNAm in CD pathogenesis. DESIGN: We generated 312 intestinal epithelial organoids (IEOs) from mucosal biopsies of 168 patients with CD (n=72), UC (n=23) and healthy controls (n=73). We performed genome-wide molecular profiling including DNAm, bulk as well as single-cell RNA sequencing. Organoids were subjected to gene editing and the functional consequences of DNAm changes evaluated using an organoid-lymphocyte coculture and a nucleotide-binding oligomerisation domain, leucine-rich repeat and CARD domain containing 5 (NLRC5) dextran sulphate sodium (DSS) colitis knock-out mouse model. RESULTS: We identified highly stable, CD-associated loss of DNAm at major histocompatibility complex (MHC) class 1 loci including NLRC5 and cognate gene upregulation. Single-cell RNA sequencing of primary mucosal tissue and IEOs confirmed the role of NLRC5 as transcriptional transactivator in the intestinal epithelium. Increased mucosal MHC-I and NLRC5 expression in adult and paediatric patients with CD was validated in additional cohorts and the functional role of MHC-I highlighted by demonstrating a relative protection from DSS-mediated mucosal inflammation in NLRC5-deficient mice. MHC-I DNAm in IEOs showed a significant correlation with CD disease phenotype and outcomes. Application of machine learning approaches enabled the development of a disease prognostic epigenetic molecular signature. CONCLUSIONS: Our study has identified epigenetically regulated intestinal epithelial MHC-I as a novel mechanism in CD pathogenesis

Patient-derived organoid biobank identifies epigenetic dysregulation of intestinal epithelial MHC-I as a novel mechanism in severe Crohn’s Disease

Peer reviewed: TrueAcknowledgements: The authors would like to thank Prof Gillian Griffith (Cambridge Institute for Medical Research, CIMR, Cambridge) for providing helpful guidance and access to tissue from OTI mice. We thank the University of Lausanne for sharing transgenic mice and Dr. A. Zenobi (IRB Bellinzona) who helped with the mouse shipment’s organization. We would like also to acknowledge Sophie Pritchard (Cellular Generation and Phenotyping core facility at the Wellcome Sanger Institute) and Dr. Heather Zecchini (CRUK Cambridge Institute Light Microscopy facility) for helping us with RNA scope and spinning disk microscopy image acquisition and analysis respectively. Furthermore, we thank Prof Louise Boyle (Department of Pathology, University of Cambridge) for advice and constructive discussions. The illustrative graphics of experiments were created with BioRender.com.Publication status: PublishedObjective: Epigenetic mechanisms, including DNA methylation (DNAm), have been proposed to play a key role in Crohn’s disease (CD) pathogenesis. However, the specific cell types and pathways affected as well as their potential impact on disease phenotype and outcome remain unknown. We set out to investigate the role of intestinal epithelial DNAm in CD pathogenesis. Design: We generated 312 intestinal epithelial organoids (IEOs) from mucosal biopsies of 168 patients with CD (n=72), UC (n=23) and healthy controls (n=73). We performed genome-wide molecular profiling including DNAm, bulk as well as single-cell RNA sequencing. Organoids were subjected to gene editing and the functional consequences of DNAm changes evaluated using an organoid-lymphocyte coculture and a nucleotide-binding oligomerisation domain, leucine-rich repeat and CARD domain containing 5 (NLRC5) dextran sulphate sodium (DSS) colitis knock-out mouse model. Results: We identified highly stable, CD-associated loss of DNAm at major histocompatibility complex (MHC) class 1 loci including NLRC5 and cognate gene upregulation. Single-cell RNA sequencing of primary mucosal tissue and IEOs confirmed the role of NLRC5 as transcriptional transactivator in the intestinal epithelium. Increased mucosal MHC-I and NLRC5 expression in adult and paediatric patients with CD was validated in additional cohorts and the functional role of MHC-I highlighted by demonstrating a relative protection from DSS-mediated mucosal inflammation in NLRC5-deficient mice. MHC-I DNAm in IEOs showed a significant correlation with CD disease phenotype and outcomes. Application of machine learning approaches enabled the development of a disease prognostic epigenetic molecular signature. Conclusions: Our study has identified epigenetically regulated intestinal epithelial MHC-I as a novel mechanism in CD pathogenesis

Inguinal Hernia: Mesh Fixation

FIBRIN GLUE MESH FIXATION UNDER LOCAL ANAESTHESIA FOR THE TREATMENT OF INGUINAL HERNIA IN ELDERLY PATIENTS R LionettF, A Cesaro 1 , E NapolitanoI, L Caruso 1 , B Neola1, M Rutigliano1, 0 P Ferulano1 iDpt. Specialistic Surgeries and Nephrology Policlinic Federico II, Naples, ITALY Introduction: Inguinal hernia repair is one of the most common operations performed in general surgery, especially among elderly patients, due to age-related loss of muscle mass and increase of co-morbidities associated with high intra-abdominal pressure. The purpose of our trial was to assess the safeness and the impact on quality of life of tension free, sutureless hernia repair technique with the use of fibrin glue under local anesthesia in elderly patients. Methods: From January 2010 to December 2012,53 male patients aged 70 and above (mean age 73.9 years) were enrolled; complicated, recurrent, scrotal hernia and ASA IV patients were excluded. Furthermore diabetic patients with glycated hemoglobin level 7% or more were ruled out for presumable neuropathy. Informed consent and data from SF36 questionnaire were collected preoperatively. The Visual Analogue Scale (VAS) for postoperative pain and a new SF36 questionnaire for overall satisfaction at one year, were administered postoperatively. Chronic pain was classified according to Cunningham's criteria. Operative time, length of hospitalization, postoperative use of nonsteroidal anti-inflammatory drugs, complications and recurrences were also assessed. Results: All patients were operated under local anaesthesia (2% Mepivacaine Cloridrate and 7,5mg!ml Ropivacaine) with light sedation; in all cases partially absorbable mesh and plug (polypropylene! polyglecaprone 25) have been implanted and fixed with I ml of fibrin glue. 50 out of 53 patients completed the 2 years follow-up, one died for not related comorbidity. Mean operative time was 54.8 minutes; 46 patients were discharged at home the same day, 5 the following day, 2 patients had to stay one more day for postoperative complications (I haematoma, I urinary retention), no major complications were observed; at two years follow-up, 2 recurrences (4%) have been observed; mean VAS score for post-operative pain, assessed at 6, 12, 24 hours and 7 days after surgery, was 4 or less for 50 (94,3%) patients, only 3 (5,6%) patients referred a score> 4. At one year follow up only 2 (4%) patients suffered of chronic postoperative pain (I mild and I moderate), no severe chronic postoperative pain has been reported. Data from pre and postoperative SF36 questionnaires, analysed by using the Student's t test, showed significant increase of the score both in the Physical Component Summery (PCM) and in the Mental Component Summary (MCS) with a p-value < 0.0001. Conclusion: Inguinal hernia repair with use of fibrin glue and partially absorbable prosthesis under local anaesthesia is a safe technique in elderly patient