Validation of miR-1228-3p as Housekeeping for MicroRNA Analysis in Liquid Biopsies from Colorectal Cancer Patients

BACKGROUND: Circulating microRNA (miRNA) analysis is a growing research field. However, it usually requires an endogenous control or housekeeping (HK) in order to normalize expression of specific miRNAs throughout different samples. Unfortunately, no adequate HK for circulating miRNA analysis is still known in the colorectal cancer (CRC) context whereas several have been suggested. Hence, our aims were to validate the previously suggested miR-1228-3p as HK for CRC studies, to compare its suitability with the widely used miR-16-5p, and to evaluate the influence of hemolysis on both miRNAs. METHODS: We analyzed by quantitative reverse-transcription polymerase chain reaction (qRT-PCR) the expression of miR-1228-3p, miR-16-5p and the spike-in cel-miR-39 in a set of 297 plasmas (92 CRC, 101 advanced adenomas -AA-, and 100 controls) and 213 serum samples (59 CRC, 74 AA and 80 controls). We also analyzed both miRNAs depending on the hemolysis degree in 7 plasmas and 31 serums. RESULTS: Levels of miR-1228-3p and miR-16-5p did not show significant differences between groups although miR-16-5p exhibited more variability in plasma and serum samples. Importantly, the combination of cel-miR-39 and miR-1228-3p was the most stable one. Moreover, we observed that miR-16-5p was significantly influenced by hemolysis in contrast with miR-1228-3p that exhibited no correlation with this confounding factor in both biofluids. CONCLUSION: MiR-1228-3p has been validated as an adequate endogenous control for circulating miRNA analysis in CRC and AA liquid biopsies

Plasma MicroRNA Signature Validation for Early Detection of Colorectal Cancer

OBJECTIVES: Specific microRNA (miRNA) signatures in biological fluids can facilitate earlier detection of the tumors being then minimally invasive diagnostic biomarkers. Circulating miRNAs have also emerged as promising diagnostic biomarkers for colorectal cancer (CRC) screening. In this study, we investigated the performance of a specific signature of miRNA in plasma samples to design a robust predictive model that can distinguish healthy individuals from those with CRC or advanced adenomas (AA) diseases. METHODS: Case control study of 297 patients from 8 Spanish centers including 100 healthy individuals, 101 diagnosed with AA, and 96 CRC cases. Quantitative real-time reverse transcription was used to quantify a signature of miRNA (miRNA19a, miRNA19b, miRNA15b, miRNA29a, miRNA335, and miRNA18a) in plasma samples. Binary classifiers (Support Vector Machine [SVM] linear, SVM radial, and SVM polynomial) were built for the best predictive model. RESULTS: Area under receiving operating characteristic curve of 0.92 (95% confidence interval 0.871-0.962) was obtained retrieving a model with a sensitivity of 0.85 and specificity of 0.90, positive predictive value of 0.94, and negative predictive value of 0.76 when advanced neoplasms (CRC and AA) were compared with healthy individuals. CONCLUSIONS: We identified and validated a signature of 6 miRNAs (miRNA19a, miRNA19b, miRNA15b, miRNA29a, miRNA335, and miRNA18a) as predictors that can differentiate significantly patients with CRC and AA from those who are healthy. However, large-scale validation studies in asymptomatic screening participants should be conducted

Elective Cancer Surgery in COVID-19-Free Surgical Pathways During the SARS-CoV-2 Pandemic: An International, Multicenter, Comparative Cohort Study.

PURPOSE: As cancer surgery restarts after the first COVID-19 wave, health care providers urgently require data to determine where elective surgery is best performed. This study aimed to determine whether COVID-19-free surgical pathways were associated with lower postoperative pulmonary complication rates compared with hospitals with no defined pathway. PATIENTS AND METHODS: This international, multicenter cohort study included patients who underwent elective surgery for 10 solid cancer types without preoperative suspicion of SARS-CoV-2. Participating hospitals included patients from local emergence of SARS-CoV-2 until April 19, 2020. At the time of surgery, hospitals were defined as having a COVID-19-free surgical pathway (complete segregation of the operating theater, critical care, and inpatient ward areas) or no defined pathway (incomplete or no segregation, areas shared with patients with COVID-19). The primary outcome was 30-day postoperative pulmonary complications (pneumonia, acute respiratory distress syndrome, unexpected ventilation). RESULTS: Of 9,171 patients from 447 hospitals in 55 countries, 2,481 were operated on in COVID-19-free surgical pathways. Patients who underwent surgery within COVID-19-free surgical pathways were younger with fewer comorbidities than those in hospitals with no defined pathway but with similar proportions of major surgery. After adjustment, pulmonary complication rates were lower with COVID-19-free surgical pathways (2.2% v 4.9%; adjusted odds ratio [aOR], 0.62; 95% CI, 0.44 to 0.86). This was consistent in sensitivity analyses for low-risk patients (American Society of Anesthesiologists grade 1/2), propensity score-matched models, and patients with negative SARS-CoV-2 preoperative tests. The postoperative SARS-CoV-2 infection rate was also lower in COVID-19-free surgical pathways (2.1% v 3.6%; aOR, 0.53; 95% CI, 0.36 to 0.76). CONCLUSION: Within available resources, dedicated COVID-19-free surgical pathways should be established to provide safe elective cancer surgery during current and before future SARS-CoV-2 outbreaks

Elective cancer surgery in COVID-19-free surgical pathways during the SARS-CoV-2 pandemic: An international, multicenter, comparative cohort study

PURPOSE As cancer surgery restarts after the first COVID-19 wave, health care providers urgently require data to determine where elective surgery is best performed. This study aimed to determine whether COVID-19–free surgical pathways were associated with lower postoperative pulmonary complication rates compared with hospitals with no defined pathway. PATIENTS AND METHODS This international, multicenter cohort study included patients who underwent elective surgery for 10 solid cancer types without preoperative suspicion of SARS-CoV-2. Participating hospitals included patients from local emergence of SARS-CoV-2 until April 19, 2020. At the time of surgery, hospitals were defined as having a COVID-19–free surgical pathway (complete segregation of the operating theater, critical care, and inpatient ward areas) or no defined pathway (incomplete or no segregation, areas shared with patients with COVID-19). The primary outcome was 30-day postoperative pulmonary complications (pneumonia, acute respiratory distress syndrome, unexpected ventilation). RESULTS Of 9,171 patients from 447 hospitals in 55 countries, 2,481 were operated on in COVID-19–free surgical pathways. Patients who underwent surgery within COVID-19–free surgical pathways were younger with fewer comorbidities than those in hospitals with no defined pathway but with similar proportions of major surgery. After adjustment, pulmonary complication rates were lower with COVID-19–free surgical pathways (2.2% v 4.9%; adjusted odds ratio [aOR], 0.62; 95% CI, 0.44 to 0.86). This was consistent in sensitivity analyses for low-risk patients (American Society of Anesthesiologists grade 1/2), propensity score–matched models, and patients with negative SARS-CoV-2 preoperative tests. The postoperative SARS-CoV-2 infection rate was also lower in COVID-19–free surgical pathways (2.1% v 3.6%; aOR, 0.53; 95% CI, 0.36 to 0.76). CONCLUSION Within available resources, dedicated COVID-19–free surgical pathways should be established to provide safe elective cancer surgery during current and before future SARS-CoV-2 outbreaks

Outcomes from elective colorectal cancer surgery during the SARS-CoV-2 pandemic

This study aimed to describe the change in surgical practice and the impact of SARS-CoV-2 on mortality after surgical resection of colorectal cancer during the initial phases of the SARS-CoV-2 pandemic

Mortality and pulmonary complications in patients undergoing surgery with perioperative SARS-CoV-2 infection: an international cohort study

Background: The impact of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) on postoperative recovery needs to be understood to inform clinical decision making during and after the COVID-19 pandemic. This study reports 30-day mortality and pulmonary complication rates in patients with perioperative SARS-CoV-2 infection. Methods: This international, multicentre, cohort study at 235 hospitals in 24 countries included all patients undergoing surgery who had SARS-CoV-2 infection confirmed within 7 days before or 30 days after surgery. The primary outcome measure was 30-day postoperative mortality and was assessed in all enrolled patients. The main secondary outcome measure was pulmonary complications, defined as pneumonia, acute respiratory distress syndrome, or unexpected postoperative ventilation. Findings: This analysis includes 1128 patients who had surgery between Jan 1 and March 31, 2020, of whom 835 (74·0%) had emergency surgery and 280 (24·8%) had elective surgery. SARS-CoV-2 infection was confirmed preoperatively in 294 (26·1%) patients. 30-day mortality was 23·8% (268 of 1128). Pulmonary complications occurred in 577 (51·2%) of 1128 patients; 30-day mortality in these patients was 38·0% (219 of 577), accounting for 81·7% (219 of 268) of all deaths. In adjusted analyses, 30-day mortality was associated with male sex (odds ratio 1·75 [95% CI 1·28–2·40], p\textless0·0001), age 70 years or older versus younger than 70 years (2·30 [1·65–3·22], p\textless0·0001), American Society of Anesthesiologists grades 3–5 versus grades 1–2 (2·35 [1·57–3·53], p\textless0·0001), malignant versus benign or obstetric diagnosis (1·55 [1·01–2·39], p=0·046), emergency versus elective surgery (1·67 [1·06–2·63], p=0·026), and major versus minor surgery (1·52 [1·01–2·31], p=0·047). Interpretation: Postoperative pulmonary complications occur in half of patients with perioperative SARS-CoV-2 infection and are associated with high mortality. Thresholds for surgery during the COVID-19 pandemic should be higher than during normal practice, particularly in men aged 70 years and older. Consideration should be given for postponing non-urgent procedures and promoting non-operative treatment to delay or avoid the need for surgery. Funding: National Institute for Health Research (NIHR), Association of Coloproctology of Great Britain and Ireland, Bowel and Cancer Research, Bowel Disease Research Foundation, Association of Upper Gastrointestinal Surgeons, British Association of Surgical Oncology, British Gynaecological Cancer Society, European Society of Coloproctology, NIHR Academy, Sarcoma UK, Vascular Society for Great Britain and Ireland, and Yorkshire Cancer Research

Casein and whey protein in vitro digestion in comparison with duodenal and jejunal in vivo digests

International audienceThe COST Action INFOGEST has developed a harmonized static in vitro digestion protocol based on physiologically relevant conditions1. This method has already proved to produce comparable results in inter-laboratory trials2. Since peptides generated during protein digestion might be biologically relevant for different physiological functions, it is important to directly compare the in vitro results towards in vivo data at peptide level. Therefore, the objective of this work was to assess the casein and whey proteins in vitro digestion according to the harmonized protocol and to match the outcomes with in vivo digests from pig duodenum (with in vitro gastric phase), and human jejunum (with the in vitro intestinal phase). Protein degradation was followed by SDS-PAGE and the HPLC-MS/MS sequenced peptides were processed using peptidomic and statistic tools. In pig duodenum, small amounts of intact caseins were present in all samples, while intact caseins were observed up to 60 minutes of gastric in vitro digestion. Regarding whey proteins, β-lactoglobulin could be detected in human jejunal digests taken at 60 minutes after ingestion, and similarly, this protein was hydrolysed after 60 min into contact with the simulated intestinal juice. The peptide profiles generated after in vitro and in vivo digestions showed clear similarities with characteristic overrepresented regions in all milk proteins. For instance, N-terminal and C-terminal domains of β-casein, and αs1-casein regions 23-30 and 183-193, were the source of abundant peptides, in both, in vitro and in vivo digests. Based on the peptide profiles, the correlation coefficients calculated between in vitro digests and the human jejunal effluents were within the range found for the different volunteers of the study. Results indicate that the harmonized in vitro protocol is a robust and simple model and constitutes a good approximation to the physiological digestion of milk proteins

Casein and whey protein in vitro digestion in comparison with duodenal and jejunal in vivo digests

Resumen del trabajo presentado a la 5th International Conference on Food Digestion, celebrada en Rennes (Francia) del 4 al 6 de abril de 2017.The COST Action INFOGEST has developed a harmonized static in vitro digestion protocol based on physiologically relevant conditions. This method has already proved to produce comparable results in inter-laboratory trials. Since peptides generated during protein digestion might be biologically relevant for different physiological functions, it is important to directly compare the in vitro results towards in vivo data at peptide level. Therefore, the objective of this work was to assess the casein and whey proteins in vitro digestion according to the harmonized protocol and to match the outcomes with in vivo digests from pig duodenum (with in vitro gastric phase), and human jejunum (with the in vítro intestinal phase). Protein.degradation was followed by SDS-PAGE and the HPLC-MS/MS sequenced peptides were processed using peptidomic and statistic tools. In pig duodenum, small amounts of intact caseins were present in all samples, while intact caseins were observed up to 60 minutes of gastric in vitro digestion. Regarding whey proteins, β-lactoglobulin could be detected in human jejunal digests taken at 60 minutes after ingestion, and similarly, this protein was hydrolysed after 60 min into contact with the simulated intestinal juice. The peptide profiles generated after in vítro and in vivo digestions showed clear similarities with characteristic overrepresented regions in all milk proteins. For instance, N-terminal and C-terminal domains of β-casein, and αs1-casein regions 23-30 and 183-193, were the source of abundant peptides, in both, in vitro and in vivo digests. Based on the peptide profiles, the correlation coefficients calculated between in vítro digests and the human jejunal effluents were within the range found for the different volunteers of the study. Results indicate that the harmonized in vítro protocol is a robust and simple model and constitutes a good approximation to the physiological digestion of milk proteins.Peer reviewe

Casein and whey protein in vitro digestion in comparison with duodenal and jejunal in vivo digests

International audienceThe COST Action INFOGEST has developed a harmonized static in vitro digestion protocol based on physiologically relevant conditions1. This method has already proved to produce comparable results in inter-laboratory trials2. Since peptides generated during protein digestion might be biologically relevant for different physiological functions, it is important to directly compare the in vitro results towards in vivo data at peptide level. Therefore, the objective of this work was to assess the casein and whey proteins in vitro digestion according to the harmonized protocol and to match the outcomes with in vivo digests from pig duodenum (with in vitro gastric phase), and human jejunum (with the in vitro intestinal phase). Protein degradation was followed by SDS-PAGE and the HPLC-MS/MS sequenced peptides were processed using peptidomic and statistic tools. In pig duodenum, small amounts of intact caseins were present in all samples, while intact caseins were observed up to 60 minutes of gastric in vitro digestion. Regarding whey proteins, β-lactoglobulin could be detected in human jejunal digests taken at 60 minutes after ingestion, and similarly, this protein was hydrolysed after 60 min into contact with the simulated intestinal juice. The peptide profiles generated after in vitro and in vivo digestions showed clear similarities with characteristic overrepresented regions in all milk proteins. For instance, N-terminal and C-terminal domains of β-casein, and αs1-casein regions 23-30 and 183-193, were the source of abundant peptides, in both, in vitro and in vivo digests. Based on the peptide profiles, the correlation coefficients calculated between in vitro digests and the human jejunal effluents were within the range found for the different volunteers of the study. Results indicate that the harmonized in vitro protocol is a robust and simple model and constitutes a good approximation to the physiological digestion of milk proteins

Nonlinear anisotropic elliptic and parabolic equations with variable exponents and L1L^1 data

International audienceWe prove existence and regularity results for distributional solutions of nonlinear elliptic and parabolic equations with general anisotropic diffusivities with variable exponents. The data are assumed to be merely integrable