Extracellular RNA Profile in Mesenteric Lymph from Exemplar Rat Models of Acute and Critical Illness.

Background: Mesenteric lymph (ML) has been implicated in the development of multiple organ dysfunction syndrome in critical illness. Extracellular RNAs play a role in cell-to-cell communication during physiological and disease processes but they are rarely studied in ML. We aimed at examining the RNA profiles of peripheral plasma, ML, and ML's extracellular vesicle (ML-EV) and triglyceride-rich lipoprotein (ML-TRL) fractions, obtained from rodent models of critical illness. Methods and Results: We collected ML for 5 hours from rodent models of critical illness [Acute Pancreatitis, Cecal Ligation and Incision (CLI), Gut Ischemia-Reperfusion (IR)] and matching Sham control rats. ML-EV and ML-TRL fractions were also isolated. RNA sequencing was performed on the RNA extracted from ML, ML-EV, ML-TRL, and plasma by using the Ion Torrent Personal Genome Machine platform. RNA sequences were searched using the Basic Local Alignment Search Tool against rat genome and RefSeq, microRNA (miRNA), genomic tRNA, functional RNA, and Genbank nucleotide databases, and the read counts were analyzed. Each sample type had a distinct RNA profile. ML contained more RNA per volume and a larger proportion of tRNA fragments than plasma. ML-EVs were enriched with miRNA, whereas ML-TRLs contained low absolute amounts of RNA. The RNA size profiles for CLI and Gut IR were different from Sham. ML carried intestinal RNAs and in a CLI model it was significantly enriched with bacterial RNA sequences. Conclusions: We found the distinct but diverse RNA profiles of ML and its compartments, and their different profiles in critical illness. Intestinal-derived small RNAs in ML may have a direct role in critical illness and utility as potential biomarkers.

Steatotic livers are susceptible to normothermic ischemia-reperfusion injury from mitochondrial Complex-I†dysfunction.

AIM: To assess the effects of ischemic preconditioning (IPC, 10-min ischemia/10-min reperfusion) on steatotic liver mitochondrial function after normothermic ischemia-reperfusion injury (IRI). METHODS: Sixty male Sprague-Dawley rats were fed 8-wk with either control chow or high-fat/high-sucrose diet inducing > 60% mixed steatosis. Three groups (n = 10/group) for each dietary state were tested: (1) the IRI group underwent 60 min partial hepatic ischemia and 4 h reperfusion; (2) the IPC group underwent IPC prior to same standard IRI; and (3) sham underwent the same surgery without IRI or IPC. Hepatic mitochondrial function was analyzed by oxygraphs. Mitochondrial Complex-I, Complex-II enzyme activity, serum alanine aminotransferase (ALT), and histological injury were measured. RESULTS: Steatotic-IRI livers had a greater increase in ALT (2476 ± 166 vs 1457 ± 103 IU/L, P < 0.01) and histological injury following IRI compared to the lean liver group. Steatotic-IRI demonstrated lower Complex-I activity at baseline [78.4 ± 2.5 vs 116.4 ± 6.0 nmol/(min.mg protein), P < 0.001] and following IRI [28.0 ± 6.2 vs 104.3 ± 12.6 nmol/(min.mg protein), P < 0.001]. Steatotic-IRI also demonstrated impaired Complex-I function post-IRI compared to the lean liver IRI group. Complex-II activity was unaffected by hepatic steatosis or IRI. Lean liver mitochondrial function was unchanged following IRI. IPC normalized ALT and histological injury in steatotic livers but had no effect on overall steatotic liver mitochondrial function or individual mitochondrial complex enzyme activities. CONCLUSION: Warm IRI impairs steatotic liver Complex-I activity and function. The protective effects of IPC in steatotic livers may not be mediated through mitochondria.

Leptin Is Associated With Persistence of Hyperglycemia in Acute Pancreatitis: A Prospective Clinical Study.

Adipokines have many homeostatic roles, including modulation of glucose metabolism, but their role in the pathophysiology of hyperglycemia associated with acute and critical illnesses in general, and acute pancreatitis (AP) in particular, is largely unknown. This study aimed to investigate the relationship between a panel of adipokines and hyperglycemia in the early course of AP, as well as the role of adipokines as predictors of AP severity.Adiponectin, leptin, omentin, resistin, and visfatin were measured on a daily basis in the first 72 hours after hospital admission. A first set of analyses was undertaken with admission glycemia stratified by severity, and a second set of analyses was undertaken based on persistence of early hyperglycemia. All of the analyses were adjusted for confounders.A total of 32 patients with AP were included in this study. None of the studied adipokines was significantly associated with glucose level on admission. Leptin was significantly (P = 0.003) increased in patients with persistent hyperglycemia. Adiponectin was significantly associated with the Acute Physiology and Chronic Health Evaluation II (APACHE II) score in patients with persistent hyperglycemia (P = 0.015), visfatin with APACHE II score in patients with persistent hyperglycemia (P = 0.014), and omentin with APACHE II score in all of the patients regardless of the presence or absence of hyperglycemia (P = 0.021).Leptin is significantly associated with persistent hyperglycemia in the early course of AP. Omentin has a potential to become an accurate predictor of AP severity.

Ghrelin and gastroparesis as early predictors of clinical outcomes in acute pancreatitis.

BACKGROUND: Impaired motor and hormonal gastrointestinal functions have been implicated in the pathogenesis of acute pancreatitis. The aim of the present study was to investigate the predictive value of the Gastroparesis Cardinal Symptom Index and serum ghrelin in the development of clinically meaningful outcomes in patients with acute pancreatitis. METHODS: This was a prospective clinical study. The Gastroparesis Cardinal Symptom Index and serum ghrelin were measured for 48 h after hospitalization. Univariate and multivariate logistic regression analyses were conducted. RESULTS: The Gastroparesis Cardinal Symptom Index total score alone on day 2 was a significant predictor of oral feeding intolerance in both unadjusted (odds ratio 1.21 (1.01-1.46), P = 0.04) and adjusted (odds ratio 1.30 (1.01-1.69), P = 0.05) analyses. Adding ghrelin to Gastroparesis Cardinal Symptom Index further improved prediction in both unadjusted (odds ratio 1.26 (1.02-1.56), P = 0.03) and adjusted (odds ratio 1.53 (1.00-2.35), P = 0.05) analyses. CONCLUSION: This pilot study demonstrates that the Gastroparesis Cardinal Symptom Index has a potential to be used as a predictor of oral feeding intolerance. Ghrelin, when combined with the Gastroparesis Cardinal Symptom Index, may further improve the predictive accuracy. These findings need to be confirmed in larger studies.

The effect of enteral nutrition on adipokines in patients with acute pancreatitis.

The mechanism behind the beneficial effects of enteral nutrition (EN) for patients with acute pancreatitis (AP) is largely unknown. Adipokines, as mediators of metabolism and inflammation, may be a possible mechanism. The study aimed to investigate the effect of EN on adipokines early in the course of AP. Patients with AP were randomised to EN or nil-by-mouth (NBM). Blood samples were taken on the first 4 d of admission and adipokine concentrations for adiponectin, leptin, omentin, resistin and visfatin were determined by ELISA assays. A linear mixed model analysis was run to determine differences in adipokine concentrations between the two study groups. A total of thirty-two patients were included in the study. Omentin concentrations were significantly higher in patients who received EN compared with NBM across the first 4 d of admission (mean difference: 11·6 (95 % CI 1·0, 22·3) ng/ml; P = 0·033). Leptin concentrations were significantly higher in patients who received EN compared with NBM after adjusting for age, sex and BMI (mean difference: 2·3 (95 % CI 0·1, 4·5) ng/ml; P = 0·037). No significant difference in adiponectin, resistin or visfatin concentrations were observed between the two study groups. EN significantly increases omentin and leptin concentrations in AP. Future research should be directed towards understanding whether these adipokines are responsible for the therapeutic benefits of EN.

The clinical relevance of obesity in acute pancreatitis: targeted systematic reviews.

BACKGROUND: With the rising prevalence of obesity, its impact on the severity and outcome of acute pancreatitis remains an important consideration when managing obese patients with acute pancreatitis. OBJECTIVE: To determine the clinical relevance of obesity in acute pancreatitis. METHODS: A series of clinically relevant questions were framed which formed the basis of our literature search using PubMed and EMBASE databases. These related to acute pancreatitis severity, systemic inflammatory response, mortality, local and systemic complications. The search was restricted to human studies. Studies were classified according to the Oxford Centre for Evidence Based Medicine levels of evidence 1 for prognostic studies. Obesity was defined according to the guidelines of the World Health Organization. In keeping with studies included the binary classification (mild and severe) of acute pancreatitis was used. RESULTS: Obesity is associated with an amplified systemic inflammatory response in acute pancreatitis and is a prognostic factor for mortality, local, systemic complications and severity in acute pancreatitis. Obesity was not found to be an independent prognostic factor for mortality and organ failure in patients with acute pancreatitis. It was evident that further studies are required to determine whether incorporating obesity into existing scoring systems improves severity prediction. Emerging evidence suggests that an obesity paradox is present in patients with acute pancreatitis. CONCLUSION: This review demonstrates that obesity has a clinically relevant impact on the course and outcome of acute pancreatitis.

Laparoscopic adrenalectomy for a rare 14-cm adrenal schwannoma.

Laparoscopic surgery for adrenal tumors is the gold standard for benign tumors; however, its role for adrenal cancer, metastases, and large suspicious lesions remains controversial. This aspect becomes clinically more important as larger incidentaloma are being detected with increasing frequency. Here, we discuss a rare case of a giant 14-cm adrenal schwannoma, which presented as an incidentaloma and was excised laparoscopically. Epidemiology, histology, and surgical treatment options were reviewed. An abdominal computerized tomography scan of a 30-year-old female weighing 130 kg revealed a large left adrenal mass. Preoperative biochemical and endocrine workup confirmed that it was nonfunctioning. The patient had a laparoscopic adrenalectomy without complication. The nodular tumor measured 145 × 100 × 80 cm in size and weighed 312 g. Histopathology showed myxoid areas and spindle cells arranged in a palisading manner. Mitoses were not observed. Tumor cells were immunohistochemically strongly positive for S-100, but negative for CD117, desmin, and muscle-specific actin. There was no evidence of malignancy. The diagnosis was of a benign schwannoma. Adrenal schwannoma is an extremely rare entity and can grow considerably in size. So far, this is the largest adrenal schwannoma reported in literature. In agreement with a growing number of publications, laparoscopic adrenalectomy can also be used for potentially malignant tumors larger than 10 cm in diameter provided the tumor does not infiltrate into other organs, conversion to open surgery is carefully considered, and resection occurs within the anatomic planes, thus ensuring the intactness of the tumor capsule.

Challenging differential diagnosis of a wild-type gastrointestinal stromal tumour (GIST) or rare reticular perineurioma of the stomach? The role for mutational analysis.

The differential diagnosis of submucosal stomach lesions includes gastrointestinal stromal tumour (GIST), leiomyoma, synovial sarcomas, perineurioma, myxoid chondrosarcoma, myoepithelial tumour and other rare mesenchymal tumours. GISTs are well-defined lesions with distinctive morphologic and histogenetic characteristics that show 95% positive staining for CD117. Differential diagnosis of wild-type GISTs can be challenging. Here, we present two stomach tumours that were operated on in our surgical department. Both presented with positive immunoreactivity for CD117. In one tumour, c-Kit mutation analysis demonstrated positivity of exon 11_c.1674_1676delGGT, thus confirming the diagnosis of a GIST. Mutational analysis of the second stomach lesion demonstrated negativity for all known c-KIT and PDGFRA exons. In situ hybridisation ruled out a synovial sarcoma. An additional immunohistochemical staining for epithelial membrane antigen eventually confirmed the diagnosis of an extremely rare reticular perineurioma in the stomach, so far reported for the second time worldwide. Both patients have not shown any signs of recurrence 2 years after surgery. The presented cases emphasise the benefits of performing a mutational analysis in difficult GISTs, including wt-GISTs, and demonstrates the importance and challenges in differentiating GISTs from other mesenchymal tumours.

The role of MYCN in the failure of MYCN amplified neuroblastoma cell lines to G1 arrest after DNA damage.

We previously reported that 3 p53 wild type (wt) MYCN amplified (MNA) neuroblastoma cell lines failed to G1 arrest after DNA damage despite induction of p53, p21(WAF1) and MDM2. We hypothesised that this was due to high MYCN expression. p53 responses to DNA damage were examined in an additional 13 p53 wt neuroblastoma cell lines. MNA was significantly associated with a failure to G1 arrest after DNA damage (p < 0.001) and higher levels of apoptosis after irradiation (p < 0.05). p21(WAF1) and hypophosphorylated (hypo) RB accumulation post irradiation were significantly lower in cell lines that failed to G1 arrest (p < 0.05). Conditional MYCN expression in non-MNA SHEP Tet21N cells did not affect the G1 arrest after irradiation. MYCN knockdown using siRNA in 3 p53 wt MNA cell lines did not restore a G1 arrest after irradiation, but increased the baseline G1 population, p21(WAF1) and hypo RB expression. MYCN siRNA also caused a G1 arrest in a p53 mutant MNA cell line. This study is the first to determine that MNA correlates with a failure to G1 arrest and attenuated p21(WAF1) induction; however MYCN expression alone is not causally responsible.