Proteomic Characterization of Circulating Molecular Perturbations Associated With Pancreatic Adenocarcinoma Following Intravenous ω-3 Fatty Acid and Gemcitabine Administration: A Pilot Study.

BACKGROUND: Administration of intravenous ω-3 fatty acid (ω-3FA) in advanced pancreatic adenocarcinoma patients receiving gemcitabine chemotherapy shows disease stabilization and improved progression-free survival. Using high-definition plasma proteomics, the underlying biological mechanisms responsible for these clinical effects are investigated. METHODS AND RESULTS: A pilot study involving plasma that was collected at baseline from 13 patients with histologically confirmed, unresectable pancreatic adenocarcinoma (baseline group) after 1-month treatment with intravenous gemcitabine and ω-3FA (treatment group) and intravenous gemcitabine only (control group) and was prepared for proteomic analysis. A 2-arm study comparing baseline vs treatment and treatment vs control was performed. Proteins were isolated from plasma with extensive immunodepletion, then digested and labeled with isobaric tandem mass tag peptide tags. Samples were then combined, fractionated, and injected into a QExactive-Orbitrap Mass-Spectrometer and analyzed on Proteome Discoverer and Scaffold with ensuing bioinformatics analysis. Selective reaction monitoring analysis was performed for verification. In total, 3476 proteins were identified. Anti-inflammatory markers (C-reactive protein, haptoglobin, and serum amyloid-A1) were reduced in the treatment group. Enrichment analysis showed angiogenesis downregulation, complement immune systems upregulation, and epigenetic modifications on histones. Pathway analysis identified direct action via the Pi3K-AKT pathway. Serum amyloid-A1 significantly reduced (P < .001) as a potential biomarker of efficacy for ω-3FA. CONCLUSIONS: This pilot study demonstrates administration of ω-3FA has potential anti-inflammatory, antiangiogenic, and proapoptotic effects via direct interaction with cancer-signaling pathways in patients with advanced pancreatic adenocarcinoma. Further studies in a larger sample size is required to validate the clinical correlation found in this preliminary study.

An ex vivo porcine spleen perfusion as a model of bacterial sepsis.

An ex vivo, porcine spleen perfusion model was established to study the early events occurring in the spleen prior to the onset of bacterial sepsis, using organs retrieved from animals slaughtered for food production. Porcine spleens were harvested from adult pigs and connected to a normothermic extracorporeal perfusion circuit. A constant perfusion of heparinized blood was performed for 6 hours. After injection of Streptococcus pneumoniae to the circuit serial samples of both blood and spleen biopsies were collected and analysed. Functionality of the perfused organs was assessed by monitoring the blood-gas parameters, flow rate and filtering capability of the organ. Interestingly, we observed full clearance of bacteria from the blood and an increase in bacterial counts in the spleen. Classical histology and immunohistochemistry on biopsies also confirmed no major damages in the organ architecture and changes in the immune cell distribution, other than the presence of clusters of pneumococci. A time-course study confirmed that each focus of infection derived from the replication of single pneumococcal cells within splenic macrophages. The model proposed - in line with the 3Rs principles - has utility in the replacement of experimental animals in infection research. Murine models are prevalently used to study pneumococcal infections, but are often not predictive for humans due to substantial differences in the immune systems of the two species. This model is designed to overcome these limitations, since porcine immunology and splenic architecture in particular, closely resemble those of humans.

Development, evaluation, and delivery of an innovative national undergraduate surgical workshop: recognition and management of the acutely unwell surgical patient.

PROBLEM: Recognition and management of acutely unwell surgical patients is an important skill to which medical students have little exposure. INTERVENTION: We present the evaluation of a novel national surgical workshop that consisted of high-fidelity simulations, lectures, case demonstrations, case discussions, and a basic surgical skills tutorial. The high-fidelity simulations re-created genuine patient encounters and were used to facilitate the acquisition of knowledge and skill in the early recognition and management of acutely unwell surgical patients. CONTEXT: The optional workshop was designed for senior medical students and delivered by surgical trainees. Students were asked to complete a 12-item evaluation questionnaire and a 26-item multiple-choice question (MCQ) quiz, which assessed their confidence; self-perceived competence; and knowledge prior to, immediately following, and 8 weeks after the workshop. Pre- and postdata were compared using student's two-tailed t test. OUTCOME: A total of 66 medical students from 6 UK universities attended, the majority of whom enjoyed the workshop (98.3%, n = 59). Participants' confidence rating (scale = 1-5) in assessing an unwell surgical patient improved from a mean of 2.5 (n = 47) to 4.4 (n = 60). Confidence in commencing initial management improved from a mean of 2.7 (n = 47) to 4.1 (n = 59). Confidence and self-perceived competence across 12 domains improved significantly following the workshop, two-tailed unpaired t test, t(22) = 8.64, p <.0001, d = 3.68. MCQ scores immediately following the workshop were a statistically significant improvement on the preworkshop MCQ scores (n = 44), paired two-tailed t test, t(43) = 7.76, p <.0001, d = 2.37, and the improvement was sustained 8 weeks following the workshop (n = 18), paired two-tailed t test, t(17) = 3.34, p =.0039, d = 1.62. LESSONS LEARNED: Feedback from students was very positive and clearly demonstrated that a workshop taught by surgical trainees improved medical students' confidence, self-perceived competence, and knowledge in the assessment and management of acutely unwell surgical patients.