Ron W. F. de Bruin

Cyclosporine interacts with mycophenolic acid by inhibiting the multidrug resistance-associated protein 2

In mycophenolate mofetil (MMF)‐treated organ transplant recipients, lower mycophenolic acid (MPA) plasma concentrations have been found in cyclosporine (CsA) compared with tacrolimus (Tac)‐based immunosuppressive regimens. We previously demonstrated that CsA decreases exposure to MPA and increases exposure to its metabolite MPA‐glucuronide (MPAG), possibly by interfering with the biliary excretion of MPAG. To elucidate the role of the multidrug resistance‐associated protein (Mrp)‐2 in the interaction between MMF and CsA, we treated three groups of 10 Mrp2‐deficient rats (TR− rat) for 6 days with either vehicle, CsA (8 mg/kg) or Tac (4 mg/kg) by oral gavage. Hereafter, co‐administration with MMF (20 mg/kg) was started in all groups and continued through day 14. The 24‐h MPA/MPAG area under the concentration‐time curve (AUC) was determined after single (day 7) and multiple MMF doses (day 14). On both study days, there were no significant differences in the mean MPA and MPAG AUC between CsA and Tac‐treated animals. We conclude that the pharmacokinetics of MMF are comparable in Mrp2‐deficient rats receiving either CsA or Tac as co‐medication. This finding suggests that CsA‐mediated inhibition of the biliary excretion of MPAG by the Mrp2 transporter is the mechanism responsible for the interaction between CsA and MMF.

Endoscopic ablation therapy for Barrett’s esophagus with high-grade dysplasia: a review

ABSTRACTBesides esophagectomy and antireflux therapy with intensive endoscopic surveillance, endoscopic ablation therapy is a new treatment modality for Barretts esophagus (BE) with high-grade dysplasia (HGD). Endoscopic surgical ablation can be performed by either a thermal, chemical, or mechanical method. This article describes the current management of patients with BE and HGD and the various methods of endoscopic ablation, including multipolar electrocoagulation, argon plasma beam coagulation, contact laser photoablation, and photodynamic therapy. It also summarizes the results of 37 patient studies, case reports, and abstracts on experimental endoscopic therapies for BE. The advantages and disadvantages of the various treatment possibilities are considered, and the future direction of the management of BE is discussed.

Short‐term dietary restriction and fasting precondition against ischemia reperfusion injury in mice

Dietary restriction (DR) extends lifespan and increases resistance to multiple forms of stress, including ischemia reperfusion injury to the brain and heart in rodents. While maximal effects on lifespan require long‐term restriction, the kinetics of onset of benefits against acute stress is not known. Here, we show that 2–4 weeks of 30% DR improved survival and kidney function following renal ischemia reperfusion injury in mice. Brief periods of water‐only fasting were similarly effective at protecting against ischemic damage. Significant protection occurred within 1 day, persisted for several days beyond the fasting period and extended to another organ, the liver. Protection by both short‐term DR and fasting correlated with improved insulin sensitivity, increased expression of markers of antioxidant defense and reduced expression of markers of inflammation and insulin/insulin‐like growth factor‐1 signaling. Unbiased transcriptional profiling of kidneys from mice subject to short‐term DR or fasting revealed a significant enrichment of signature genes of long‐term DR. These data demonstrate that brief periods of reduced food intake, including short‐term daily restriction and fasting, can increase resistance to ischemia reperfusion injury in rodents and suggest a rapid onset of benefits of DR in mammals.

Angiotensin-(1–7) and the G Protein-Coupled Receptor Mas Are Key Players in Renal Inflammation

Angiotensin (Ang) II mediates pathophysiologial changes in the kidney. Ang-(1–7) by interacting with the G protein-coupled receptor Mas may also have important biological activities. In this study, renal deficiency for Mas diminished renal damage in models of renal insufficiency as unilateral ureteral obstruction and ischemia/reperfusion injury while the infusion of Ang-(1–7) to wild-type mice pronounced the pathological outcome by aggravating the inflammatory response. Mas deficiency inhibited NF-κB activation and thus the elevation of inflammation-stimulating cytokines, while Ang-(1–7) infusion had proinflammatory properties in experimental models of renal failure as well as under basal conditions. The Ang-(1–7)-mediated NF-κB activation was Mas dependent but did not involve Ang II receptors. Therefore, the blockade of the NF-κB-activating properties of the receptor Mas could be a new strategy in the therapy of failing kidney.

5-Aminolaevulinic acid-induced protoporphyrin IX accumulation in tissues: Pharmacokinetics after oral or intravenous administration

UNLABELLED In this study, the biodistribution of 5-aminolaevulinic acid (ALA) and accumulation of protoporphyrin IX (PpIX) in rats have been examined. Two groups of 21 WAG/Rij rats are given 200 mg/kg ALA orally or intravenously. Six rats serve as controls. At 1, 2, 3, 4, 6, 12 and 24 h after ALA administration, ALA and porphyrin concentrations are measured in 18 tissues and fluids. Liver enzymes and renal-function tests are measured to determine ALA toxicity. In both groups ALA concentration is highest in kidney, bladder and urine. After oral administration, high concentrations are also found in duodenal aspirate and jejunum. Mild, short-lasting elevation of creatinine is seen in both treatment groups. Porphyrins, especially PpIX, accumulate mainly in duodenal aspirate, jejunum, liver and kidney (> 10 nmol/g tissue), less in oesophagus, stomach, colon, spleen, bladder, heart, lung and nerve (2-10 nmol/g tissue), and only slightly in plasma, muscle, fat, skin and brain (< 2 nmol/g tissue). In situ synthesis of porphyrins rather than enterohepatic circulation contributes to the PpIX accumulation. Confocal laser scanning microscopy shows selective porphyrin fluorescence in epithelial layers. Peak levels and total production of porphyrins are equal after oral and intravenous ALA administration. IN CONCLUSION administration of 200 mg/kg ALA results in accumulation of photosensitive concentrations of PpIX, 1 to 6 h after ALA administration, in all tissues except muscle, fat, skin and brain. Knowledge of the time-concentration relationship should be helpful in selecting dosages, routes of administration and timing of ALA photodynamic therapy.

Impact of temperature and humidity of carbon dioxide pneumoperitoneum on body temperature and peritoneal morphology.

BACKGROUND The insufflation of cold gas during laparoscopic surgery exposes patients to the risk for hypothermia. The objectives of this study were to investigate whether heating or humidification of insufflation gas could prevent peroperative hypothermia in a rat model, and to assess whether the peritoneum was affected by heating or humidification of the insufflation gas. METHODS Rats were exposed to insufflation with either cold, dry carbon dioxide CO2 (group I); cold, humidified CO2 (group II); warm, dry CO2 (group III); or warm, humidified CO2 (group IV); another group underwent gasless laparoscopy (group V). Core temperature and intraperitoneal temperature were registered in all animals during 120 minutes. Specimens of the parietal peritoneum were taken directly after desufflation and 2 and 24 hours after the procedure. All specimens were analyzed with scanning electron microscopy (SEM). RESULTS During the 120-minute study period, core temperature and intraperitoneal temperature were significantly reduced in groups I, II, and III. In the animals that underwent warm, humidified insufflation (group IV) and the gasless controls (group V), intraoperative hypothermia did not develop. At SEM, retraction and bulging of mesothelial cells and exposure of the basal lamina were seen in the four insufflation groups (groups I-IV) and also in the gasless controls (group V). CONCLUSION Insufflation with cold, dry CO2 may lower the body temperature during laparoscopic surgery. Hypothermia can be prevented by both heating and humidifying the insufflation gas. Changes of the peritoneal surface occur after CO2 insufflation, despite heating or humidifying, and also after gasless surgery.

Mesenchymal Stem Cells Induce an Inflammatory Response After Intravenous Infusion

Mesenchymal stem cells (MSCs) have potent immunosuppressive effects in vitro and are considered as a therapeutic option for autoimmune disease and organ transplantation. While MSCs show beneficial effects on immune disease progression and transplant survival in animal models, the immunomodulatory mechanisms involved are largely unknown. In the present study, we show that intravenously infused C57BL/6- green fluorescent protein (GFP) MSCs home to the lungs in C57BL/6 recipient mice and induce an inflammatory response. This response was characterized by increased mRNA expression of monocyte chemoattractant protein-1 (MCP1), IL1-β, and TNF-α and an increase in macrophages in lung tissue 2 h after MSC infusion. Simultaneously, serum levels of proinflammatory IL6, CXCL1, and MCP1 protein increased, demonstrating systemic immune activation after MSC infusion. In liver tissue, no C57BL/6-GFP MSCs were detected, but MCP1 and TNF-α mRNA levels peaked 4 h after MSC infusion. The expression of the anti-inflammatory cytokines TGF-β, IL4, and IL10 was only marginally affected. Nevertheless, 3 days after MSC infusion, animals developed a milder inflammatory response to lipopolysaccharides. Our results suggest that the in vivo immunomodulatory effects of MSCs originate from an inflammatory response that is induced by the infusion of MSCs, which is followed by a phase of reduced immune reactivity.

Systematic Review and Meta‐Analysis of the Impact of Computed Tomography–Assessed Skeletal Muscle Mass on Outcome in Patients Awaiting or Undergoing Liver Transplantation

Liver transplant outcome has improved considerably as a direct result of optimized surgical and anesthesiological techniques and organ allocation programs. Because there remains a shortage of human organs, strict selection of transplant candidates remains of paramount importance. Recently, computed tomography (CT)‐assessed low skeletal muscle mass (i.e. sarcopenia) was identified as a novel prognostic parameter to predict outcome in liver transplant candidates. A systematic review and meta‐analysis on the impact of CT‐assessed skeletal muscle mass on outcome in liver transplant candidates were performed according to the Preferred Reporting Items for Systematic Reviews and Meta‐Analysis guidelines. Nineteen studies, including 3803 patients in partly overlapping cohorts, fulfilled the inclusion criteria. The prevalence of sarcopenia ranged from 22.2% to 70%. An independent association between low muscle mass and posttransplantation and waiting list mortality was described in 4 of the 6 and 6 of the 11 studies, respectively. The pooled hazard ratios of sarcopenia were 1.84 (95% confidence interval 1.11–3.05, p = 0.02) and 1.72 (95% confidence interval 0.99–3.00, p = 0.05) for posttransplantation and waiting list mortality, respectively, independent of Model for End‐stage Liver Disease score. Less‐consistent evidence suggested a higher complication rate, particularly infections, in sarcopenic patients. In conclusion, sarcopenia is an independent predictor for outcome in liver transplantation patients and could be used for risk assessment.

Long-Term Impact of Pneumoperitoneum Used for Laparoscopic Donor Nephrectomy on Renal Function and Histomorphology in Donor and Recipient Rats

ObjectiveTo investigate the long-term impact of pneumoperitoneum used for laparoscopic donor nephrectomy on renal function and histomorphology in donor and recipient. Summary Background DataLaparoscopic donor nephrectomy has the potential to increase the number of living kidney donations by reducing donor morbidity. However, function of laparoscopically procured kidneys might be at risk due to ischemia as a consequence of elevated intra-abdominal pressure during laparoscopy. MethodsIn experiment 1, 30 Brown Norway rats were randomized to three procedures: 2 hours of CO2 insufflation, 2 hours of helium insufflation, and 2 hours of gasless laparoscopy. After this, a unilateral nephrectomy was performed in all animals. Another six rats were used as controls. In experiment 2, 36 donor Brown Norway rats were subjected to a similar insufflation protocol, but after nephrectomy a syngeneic renal transplantation was performed. All rats had a follow-up period of 12 months. Urine and blood samples were collected each month for determination of renal function. After 1 year, donor and recipient kidneys were removed for histomorphologic and immunohistochemical analysis. ResultsIn donors as well as in recipients, no significant changes in serum creatinine, proteinuria, or glomerular filtration rate were detected between the CO2, the helium, and the gasless control group after 1 year. No histologic abnormalities due to abdominal gas insufflation were found. Immunohistochemical analysis did not show significant differences in the number of infiltrating cells (CD4, CD8, ED1, OX62, and OX6) and adhesion molecule expression (ICAM-1) between the three groups. ConclusionsAbdominal gas insufflation does not impair renal function in the donor 1 year after LDN. One year after transplantation, no differences in renal function or histomorphology were detected between kidney grafts exposed to either pneumoperitoneum or a gasless procedure.

Conversion of metaplastic Barrett’s epithelium into post-mitotic goblet cells by γ-secretase inhibition

SUMMARY Barrett’s esophagus (BE) affects approximately 2% of the Western population and progresses to esophageal adenocarcinoma (EAC) in 0.5% of these patients each year. In BE, the stratified epithelium is replaced by an intestinal-type epithelium owing to chronic gastroduodenal reflux. Since self-renewal of intestinal crypts is driven by Notch signaling, we investigated whether this pathway was active in the proliferative crypts of BE. Immunohistochemistry confirmed the presence of an intact and activated Notch signaling pathway in metaplastic BE epithelium, but not in the normal human esophagus. Similar observations were made in two well-known human Barrett’s-derived EAC cell lines, OE33 and SKGT-5. We then sought to investigate the effects of Notch inhibition by systemic treatment with a γ-secretase inhibitor in a well-validated rodent model for BE. As we have shown previously in normal intestinal epithelium, Notch inhibition converted the proliferative Barrett’s epithelial cells into terminally differentiated goblet cells, whereas the squamous epithelium remained intact. These data imply that local application of γ-secretase inhibitors may present a simple therapeutic strategy for this increasingly common pre-malignant condition.