A Chimeric IL-15/IL-15Rα Molecule Expressed on NFκB-Activated Dendritic Cells Supports Their Capability to Activate Natural Killer Cells.

Natural killer (NK) cells, members of the innate immune system, play an important role in the rejection of HLA class I negative tumor cells. Hence, a therapeutic vaccine, which can activate NK cells in addition to cells of the adaptive immune system might induce a more comprehensive cellular response, which could lead to increased tumor elimination. Dendritic cells (DCs) are capable of activating and expanding NK cells, especially when the NFκB pathway is activated in the DCs thereby leading to the secretion of the cytokine IL-12. Another prominent NK cell activator is IL-15, which can be bound by the IL-15 receptor alpha-chain (IL-15Rα) to be transpresented to the NK cells. However, monocyte-derived DCs do neither secrete IL-15, nor express the IL-15Rα. Hence, we designed a chimeric protein consisting of IL-15 and the IL-15Rα. Upon mRNA electroporation, the fusion protein was detectable on the surface of the DCs, and increased the potential of NFκB-activated, IL-12-producing DC to activate NK cells in an autologous cell culture system with ex vivo-generated cells from healthy donors. These data show that a chimeric IL-15/IL-15Rα molecule can be expressed by monocyte-derived DCs, is trafficked to the cell surface, and is functional regarding the activation of NK cells. These data represent an initial proof-of-concept for an additional possibility of further improving cellular DC-based immunotherapies of cancer.

Bovine colostrum improves neonatal growth, digestive function, and gut immunity relative to donor human milk and infant formula in preterm pigs.

Mother's own milk is the optimal first diet for preterm infants, but donor human milk (DM) or infant formula (IF) is used when supply is limited. We hypothesized that a gradual introduction of bovine colostrum (BC) or DM improves gut maturation, relative to IF during the first 11 days after preterm birth. Preterm pigs were fed gradually advancing doses of BC, DM, or IF (3-15 ml·kg(-1)·3 h(-1), n = 14-18) before measurements of gut structure, function, microbiology, and immunology. The BC pigs showed higher body growth, intestinal hexose uptake, and transit time and reduced diarrhea and gut permeability, relative to DM and IF pigs (P < 0.05). Relative to IF pigs, BC pigs also had lower density of mucosa-associated bacteria and of some putative pathogens in colon, together with higher intestinal villi, mucosal mass, brush-border enzyme activities, colonic short chain fatty acid levels, and bacterial diversity and an altered expression of immune-related genes (higher TNFα, IL17; lower IL8, TLR2, TFF, MUC1, MUC2) (all P < 0.05). Values in DM pigs were intermediate. Severe necrotizing enterocolitis (NEC) was observed in >50% of IF pigs, while only subclinical intestinal lesions were evident from DM and BC pigs. BC, and to some degree DM, are superior to preterm IF in stimulating gut maturation and body growth, using a gradual advancement of enteral feeding volume over the first 11 days after preterm birth in piglets. Whether the same is true in preterm infants remains to be tested.

Phase angle as a prognostic marker after percutaneous endoscopic gastrostomy (PEG) in a prospective cohort study.

OBJECTIVE: The phase angle identifies changes in tissue's electrical properties assessed by bioelectrical impedance measurement and it can predict prognosis in some conditions. Percutaneous endoscopic gastrostomy (PEG) is commonly used in patients with severe nutritional problems, but there is a need to improve the clinical decision-making for using PEG. We examined if a decreased phase angle predicts complications, short-term mortality (within 60 days of PEG insertion), or inflammatory markers (high C-reactive protein [CRP] levels or low albumin levels) following PEG insertion. MATERIAL AND METHODS: The phase angle was assessed from body resistance and reactance as measured by bioelectrical impedance in 131 patients admitted for PEG. Anthropometrics and clinical biochemical measures were collected at the time of PEG insertion, while complications and mortality were assessed at clinical follow-ups. Multivariable logistic regression analysis provided odds ratios (ORs) with 95% confidence intervals (CIs) adjusted for sex, age, body mass index, and comorbidity. RESULTS: A decreased phase angle did not statistically significantly increase the probability of acute complications or short-term mortality, but predicted increased inflammatory markers (CRP ≥10 mg/L [OR 1.63, 95% CI 1.02-2.60], albumin <30 g/L [OR 2.10, 95% CI 1.24-3.57] and a combination of CRP ≥10 mg/L and albumin <30 g/L [OR 3.06, 95% CI 1.51-6.19]). CONCLUSIONS: A decreased phase angle did not predict acute complications or short-term mortality after PEG insertion, but predicted increased levels of inflammatory markers.

Risk factors for weight loss among patients surviving 5 years after esophageal cancer surgery.

BACKGROUND: This study aimed to identify factors influencing postoperative weight loss ≥15 % in long-term survivors after esophageal cancer surgery. METHODS: A population-based study was conducted in Sweden between 2001 and 2005, with regular follow-up for 5 years. Current weight, weight at operation, and average weight were assessed and dichotomized as weight loss of ≥15 %. Logistic regression estimated relative risks of weight loss between pre- and 5 years postoperatively, expressed as odds ratios (ORs) with 95 % confidence intervals (CIs). Statistically significant differences in nutritional symptoms between weight loss groups were analyzed using linear regression models to likewise test as risk factors. Body mass index (BMI) at operation (< or ≥25), sex, and preoperative weight loss (< or ≥10 %) were tested as risk factors. Nutritional symptoms were selected from the European Organisation for Research and Treatment of Cancer (EORTC) QLQ-C30 and QLQ-OES18, including fatigue, nausea and vomiting, dyspnoea, appetite loss, diarrhea, and dysphagia, eating difficulties, reflux and pain. RESULTS: A total of 117 patients were included. Patients with preoperative BMI ≥25 were at a threefold increased risk (OR 3.2; 95 % CI 1.4-7.3) for postoperative weight loss of ≥15 % 5 years after operation. Moreover, eating difficulties, pain, fatigue, nausea and vomiting, and appetite loss were clinically relevant and statistically significantly worse symptoms experienced among those with a weight loss of ≥15 % (all at p < 0.05). CONCLUSIONS: Overweight at the time of operation is associated with postoperative weight loss from a long-term perspective. Several nutritional symptoms are associated with weight loss of >15 % 5 years postoperatively.

A high amount of dietary zinc changes the expression of zinc transporters and metallothionein in jejunal epithelial cells in vitro and in vivo but does not prevent zinc accumulation in jejunal tissue of piglets.

High dietary zinc concentrations are used to prevent or treat diarrhea in piglets and humans, but long-term adaptation to high zinc supply has yet not been assessed. Intestinal zinc uptake is facilitated through members of zinc transporter families SLC30 (ZnT) and SLC39 (ZIP). Whereas in rodents, regulation of zinc homeostasis at low or adequate zinc supply has been described, such mechanisms are unclear in piglets. A total of 54 piglets were fed diets containing 57 [low dietary zinc (LZn)], 164 [normal dietary zinc (NZn)], or 2425 [high dietary zinc (HZn)] mg/kg dry matter zinc. After 4 wk, 10 piglets/group were killed and jejunal tissues taken for analysis of zinc transporters SLC30A1 (ZnT1), SLC30A2 (ZnT2), SLC30A5 (ZnT5), SLC39A4 (ZIP4), divalent metal transporter 1 (DMT1), and metallothionein-1 (MT). Weight gain was higher (P < 0.05) in pigs fed HZn than in the LZn and NZn groups during the first 2 wk. Food intake did not differ between groups. The digesta and jejunal tissue zinc concentrations were higher (P < 0.05) in the HZn pigs than in NZn and LZn pigs. Expression of ZnT1 was higher (P < 0.05) and ZIP4 lower (P < 0.05) in HZn pigs than in the 2 other groups, whereas expression of ZnT5 and DMT1 did not differ between treatments. Expression of ZnT2 was lower (P < 0.05) in the LZn group than in the HZn and NZn groups. The mRNA expression and protein abundance of MT was higher (P < 0.05) in the HZn group than in the NZn and LZn groups. Studies with intestinal porcine cell line intestinal epithelial cell-J2 confirmed the dose-dependent downregulation of ZIP4 and upregulation of ZnT1 and MT (P < 0.05) with increasing zinc concentration within 24 h. In conclusion, high dietary zinc concentrations increase intracellular zinc, promote increased zinc export from intestinal tissues into extracellular compartments, and decrease zinc uptake from the gut lumen. The adaptive process appears to be established within 24 h; however, it does not prevent tissue zinc accumulation.

Dietary crystalline common-, micro-, nanoscale and emulsified nanoscale sitosterol reduce equally the cholesterol pool in guinea pigs, but varying nanosystems result in different sterol concentrations in serosal jejunum.

Due to hypocholesterolemic effects, sitosterol is used in functional foods and nanoscale dispersions. To investigate the influence of dietary sitosterol on sterol concentrations in Dunkin Hartley guinea pigs, seven groups consisting of eight animals each were fed either a basal diet (BD), a high-cholesterol diet (HC) or a high-cholesterol diet supplemented with crystalline commonscale (CCS), microscale (CMS, low-dosed: CMLS), nanoscale (CNS) or emulsified nanoscale (ENS) sitosterol. When compared to HC group, all sitosterol formulations decreased liver cholesterol concentrations. No differences in cholesterol or sitosterol concentration were found in plasma and liver between CCS, CMS, CNS, and ENS groups. Apparent cholesterol digestibility decreased by increasing crystalline microscale sitosterol doses. Despite a lower sitosterol intake, ENS group had higher serosal sitosterol concentrations in jejunum than CNS group. To elucidate an impact of the sitosterol nanosystem on gut tissues further studies are necessary. FROM THE CLINICAL EDITOR: In this study, the use of sitosterols in a rat model led to contradicting conclusions regarding their ability to reduce cholesterol levels efficiently in guinea pigs, suggesting that more preclinical data is needed before this method could become applicable to human studies.

Fermentable fiber ameliorates fermentable protein-induced changes in microbial ecology, but not the mucosal response, in the colon of piglets.

Dietary inclusion of fermentable carbohydrates (fCHO) is reported to reduce large intestinal formation of putatively toxic metabolites derived from fermentable proteins (fCP). However, the influence of diets high in fCP concentration on epithelial response and interaction with fCHO is still unclear. Thirty-two weaned piglets were fed 4 diets in a 2 × 2 factorial design with low fCP/low fCHO [14.5% crude protein (CP)/14.5% total dietary fiber (TDF)]; low fCP/high fCHO (14.8% CP/16.6% TDF); high fCP low fCHO (19.8% CP/14.5% TDF); and high fCP/high fCHO (20.1% CP/18.0% TDF) as dietary treatments. After 21-23 d, pigs were killed and colon digesta and tissue samples analyzed for indices of microbial ecology, tissue expression of genes for cell turnover, cytokines, mucus genes (MUC), and oxidative stress indices. Pig performance was unaffected by diet. fCP increased (P < 0.05) cell counts of clostridia in the Clostridium leptum group and total short and branched chain fatty acids, ammonia, putrescine, histamine, and spermidine concentrations, whereas high fCHO increased (P < 0.05) cell counts of clostridia in the C. leptum and C. coccoides groups, shifted the acetate to propionate ratio toward acetate (P < 0.05), and reduced ammonia and putrescine (P < 0.05). High dietary fCP increased (P < 0.05) expression of PCNA, IL1ß, IL10, TGFß, MUC1, MUC2, and MUC20, irrespective of fCHO concentration. The ratio of glutathione:glutathione disulfide was reduced (P < 0.05) by fCP and the expression of glutathione transferase was reduced by fCHO (P < 0.05). In conclusion, fermentable fiber ameliorates fermentable protein-induced changes in most measures of luminal microbial ecology but not the mucosal response in the large intestine of pigs.

Complications after percutaneous endoscopic gastrostomy in a prospective study.

BACKGROUND: Insertion of a percutaneous endoscopic gastrostomy (PEG) is an increasingly common procedure in patients with nutritional needs and dysphagia. Better knowledge of rates and patterns of complications after PEG might influence decision-making. MATERIAL AND METHODS: The objective was to prospectively evaluate the rate of six pre-defined complications (leakage, diarrhea, constipation, abdominal pain, fever and peristomal infection) and mortality occurring within 2 months after PEG in an unselected sample of patients. All patients (n = 484) who had a PEG inserted at the hospital during the study period were included. Kaplan-Meier curves were used to estimate mortality over the first 60 days following PEG and Fisher's exact test was used to test equality of proportions. RESULTS: Of the 484 patients included, 85 (18%) died within 2 months after PEG insertion. The risk of early mortality was higher in the group with neurological disease than in the group with a tumor as indication (p < 0.001). After excluding mortality, the overall complication rates at 2 weeks and 2 months were 39% and 27%, respectively. The most common complications within 2 weeks were abdominal pain (13%), peristomal infection (11%), diarrhea (11%) and leakage (10%). At 2 months the most frequent complications were diarrhea (10%), leakage (8%) and peristomal infection (6%). CONCLUSIONS: In the short-term perspective, there is a substantial risk of complications, including mortality, after PEG insertion. This should be considered during clinical decision-making and when informing the patients and caregivers.

Patients' perspectives of living with a percutaneous endoscopic gastrostomy (PEG).

BACKGROUND: Since enteral nutrition therapy is the preferred nutritional support for dysphagic patients with a range of diagnoses, PEG has become part of traditional care. However, enteral nutrition with PEG transfers treatment responsibility and activity to the patients and their carers, so the advantages should be discussed. The aim of this study was therefore to investigate patients' experience of living with a percutaneous endoscopic gastrostomy (PEG) in order to increase the understanding of patients' need for support. METHOD: In a prospective study at Karolinska University Hospital in Sweden, data were collected consecutively at the time of PEG and two months later using a study-specific questionnaire about each patient's experience of living with a PEG. Fishers exact test was used to test for statistically significant difference at five per cent level. RESULTS: There were 104 responders (response rate of 70%). Women felt more limited in daily activity compared to men (p = 0.004). Older patients experienced a more limited ability to influence the number of feeding times compared to younger (p = 0.026). Highly educated patients found feeding more time-consuming (p = 0.004). Patients with a cancer diagnosis reported that the PEG feeding interfered with their oral feeding more than patients with a neurological disease (p = 0.009). Patients mostly contacted the PEG outpatient clinic with problems regarding their PEG, and were mainly assisted by their spouse rather than district nurses. CONCLUSIONS: PEG feeding is time-consuming and interferes with daily life. Although 73% was satisfied, patients' experiences of living with a PEG may be dependent on age, sex, education and diagnosis. Spouses are the main carers for PEG patients at home, and patients prefer to go to the PEG outpatient clinic for help if problems occur.

Twelve tips for conducting a postgraduate course on study design and study protocol writing for the medical profession.

BACKGROUND: It is desirable to use a methodological approach of the highest scientific quality when planning and conducting valid research. However, several studies have shown that the biostatistical and epidemiological knowledge of academically active health care professionals is far below the desired level. AIM: To outline an approach to conducting a postgraduate course in clinical study design and study protocol writing. METHOD: We offer 12 tips based on critical reflection of our experience of conducting and continuously improving a course in study design repeatedly over several years, combined with evidence from the literature on pedagogical approaches. RESULT: The tips are organized in chronological order so that a course director will be able to develop a similar course. CONCLUSION: Combining the results of our critical reflection with evidence from the literature has allowed us to develop a successful approach to running courses in study design and study protocol writing. We hope that our extensive experience in conducting this course, as reflected by the 12 tips, will support and inspire others planning similar courses.

Influence of Fetomaternal Microchimerism on Maternal NK Cell Reactivity against the Child's Leukemic Blasts.

Persistence of fetal cells in the circulation of the mother (fetal microchimerism, FM) is associated with increased survival and reduced relapse of children with leukemia receiving a haploidentical hematopoietic stem cell transplantation (hHSCT). NK cells play an important role in maternal tolerance towards the unborn child. In this study, 70 mother-child pairs were prospectively analyzed for the occurrence of FM, KIR genotype and HLA-C type. We found that occurrence and level of FM were influenced by three maternal genetic factors: presence of an HLA-C1 allele, absence of KIR2DL3 and presence of a cen-B/B motif. Furthermore, an HLA-C match between mother and child favored persistence of FM. NK cells from FM+ mothers showed a 40% higher specific degranulation against their filial leukemic blasts than NK cells from FM- mothers, suggesting the presence of educated maternal NK cells. Nevertheless, cytotoxicity of parental NK cells against filial leukemic blasts was independent of KIR genetics (haplotype, B content score, centromeric and telomeric KIR gene regions) and independent of FM, indicating that additional immune effector mechanisms contribute to the beneficial effect of persisting FM in hHSCT.

Albumin and C-reactive protein levels predict short-term mortality after percutaneous endoscopic gastrostomy in a prospective cohort study.

BACKGROUND: Percutaneous endoscopic gastrostomy (PEG) is a procedure with many complications that sometimes can be devastating. To give better advice to patients referred for PEG regarding risk of complications, important risk factors should be known. OBJECTIVE: To evaluate whether age, body mass index, albumin levels, C-reactive protein (CRP) levels, indication for PEG, and comorbidity influence the risk of mortality or peristomal infection after PEG insertion. DESIGN: Prospective cohort study from 2005 to 2009. Follow-up 14 days after PEG. SETTING: University hospital. PATIENTS: This study involved 484 patients referred for PEG. INTERVENTION: PEG. MAIN OUTCOME MEASUREMENTS: Mortality within 30 days and peristomal infection within 14 days after PEG insertion. All risk estimates were calculated with 95% CIs and adjusted for confounding. RESULTS: Among 484 patients, 58 (12%) died within 30 days after PEG insertion. Albumin <30 g/L (hazard ratio [HR], 3.46; 95% CI, 1.75-6.88), CRP ≥10 (HR, 3.47; 95% CI, 1.68-7.18), age ≥65 years (HR, 2.26; 95% CI, 1.20-4.25) and possibly body mass index <18.5 (HR, 2.04; 95% CI, 0.97-4.31) were associated with increased mortality. Patients with a combination of low albumin and high CRP levels had a mortality rate of 20.5% compared with 2.6% among patients with normal values, rendering an over 7-fold increased adjusted risk of mortality (HR, 7.45; 95% CI, 2.62-21.19). LIMITATIONS: Missing data in some study variables. Although the sample size was large, weaker associations could not be established. CONCLUSION: The combination of low albumin and high CRP levels indicates a substantially increased short-term mortality risk after PEG, which should be considered in decision making.

Prevalence and intensity of dumping symptoms and their association with health-related quality of life following surgery for oesophageal cancer.

BACKGROUND & AIMS: This study aimed to investigate the prevalence and intensity of symptoms of dumping syndrome (early and late) experienced by oesophageal cancer survivors one year after surgery and their association with health related quality of life (HRQL). METHODS: A prospective cohort study of patients who underwent surgery for oesophageal cancer in Sweden from January 2013 to April 2018, included at one year after surgery with follow-up at 1.5 years. Common symptoms of dumping syndrome were the exposure, classified as early and late onset, further divided into 'moderate' or 'severe' based on symptom intensity, and no dumping symptoms (reference group). The primary outcome was mean summary score of HRQL, and secondary outcomes were global quality of life, physical, role, emotional, cognitive and social function measured using the EORTC QLQ-C30 1.5 years after surgery. An ANCOVA model, adjusted for potential confounders was used to study the association between dumping symptoms and HRQL, presented as mean score differences (MD) with 95% confidence intervals (CI). RESULTS: Among 188 patients, moderate early dumping symptoms was experienced by 45% and severe early dumping by 9%. Moderate late dumping symptoms was reported by 13%, whereas 5% reported severe late dumping symptoms. Severe early dumping symptoms was associated with worse HRQL in 4 out of 7 aspects with worse global quality of life (MD -16, 95% CI: -27 to -4) and social function (MD -17, 95% CI: -32 to -3), which showed clinically large differences compared to having no such symptoms. Patients with moderate late dumping symptoms reported poorer HRQL in 6 out of 7 aspects compared to those with no dumping symptoms. Cognitive function (MD -27, 95% CI: -47 to -7) and emotional function (MD -24, 95% CI: -47 to -2) were significantly declined (clinically large relevance) in those with severe late dumping symptoms. CONCLUSIONS: Patients who have undergone curative treatment for oesophageal cancer experience reduced HRQL from early and late dumping symptoms at one year after surgery that indicate clear implications for clinical routine. Medical support and additional dietary counselling are required as potential ways to alleviate dumping symptoms on clinical repercussions.

Body composition evaluation with computed tomography: Contrast media and slice thickness cause methodological errors.

OBJECTIVE: Although computed tomography (CT) is frequently used to determine body composition, the effects of using different CT protocols is not well known. The aim of this study was to determine whether contrast media phase, radiation dose, and slice thickness in CT affect body composition segmentation. METHODS: Clinically indicated perfusion CTs of the upper abdomen in 20 patients (seven women) between 40 and 87 y of age with high suspicion of hepatocellular carcinoma were analyzed retrospectively. Axial images from the L3 level with varying imaging delay were reconstructed after contrast media injection (18 images per patient), slice thickness (5 images, 2-10 mm), and radiation dose (4 images with one-third to four-thirds of standard dose). Muscle and fat areas were segmented semiautomatically by drawing regions of interests and using established cutoff thresholds. Skeletal muscle index (SMI), steatotic muscle area, and adipose tissue index, as well as muscle attenuation and fat attenuation, were evaluated. RESULTS: Average SMI increased by up to 2.8% after contrast media injection. Steatotic muscle area decreased by ≤13.8%, and adipose tissue index decreased by ≤6.5%. Muscle attenuation increased after contrast media injection, whereas fat attenuation decreased (all P < 0.001). SMI decreased by 1.9% on average when increasing slice thickness from 2 to 10 mm. Steatotic muscle area increased by ≤3.3%, and adipose tissue index increased by ≤1.5% (all P < 0.05). Muscle attenuation did not change significantly with reconstruction thickness. Radiation dose had no effect on estimated area of spinal muscle, fatty spinal muscle, or visceral fat. CONCLUSIONS: Contrast media have a strong effect on the evaluation of body composition, whereas the influence of slice thickness is less pronounced. Radiation dose can be reduced by ≥66% without significantly affecting segmentation.

Effect of fecal microbiota transplantation route of administration on gut colonization and host response in preterm pigs.

This study examined gut colonization patterns and host responses to fecal microbiota transplantation (FMT) by different administration routes after preterm birth. In two separate experiments, cesarean-delivered, preterm pigs were administered combined oral + rectal, or exclusively rectal donor feces, and compared with saline controls. After 5 days, stomach and colon bacterial compositions were determined by 16S rRNA gene amplicon sequencing, and organic acid metabolites measured. Further, gut pathology, mucosa bacterial adherence, and goblet cell density were assessed. FMT increased the relative abundance of obligate anaerobes in the colon without affecting total bacterial load. Bacteroides colonized recipients despite low abundance in the donor feces, whereas highly abundant Prevotella and Ruminococcaceae did not. Further, FMT changed carbohydrate metabolism from lactate to propionate production thereby increasing colonic pH. Besides, FMT preserved goblet cell mucin stores and reduced necrotizing enterocolitis incidence. Only rectal FMT increased the stomach-to-colon pH gradient and resistance to mucosa bacterial adhesion. Conversely, oral + rectal FMT increased bacterial adhesion, internal organ colonization, and overall mortality. Our results uncovered distinctions in bacterial colonization patterns along the gastrointestinal tract, as well as host tolerability between oral and rectal FMT administration in preterm newborns. Besides, FMT showed the potential to prevent necrotizing enterocolitis.

Nutrition Impact Symptoms Are Prognostic of Quality of Life and Mortality after Surgery for Oesophageal Cancer.

We aimed to clarify the influence of nutritional problems after surgery for oesophageal cancer on functional health related quality of life (HRQOL) and survival. A prospective nationwide cohort of oesophageal cancer patients operated 2001⁻2005 in Sweden with 6 months postoperative follow up was used. Nutritional problems were categorized as low/moderate/severe/very severe based on weight loss and nutrition impact symptoms. An ANCOVA model calculated mean score differences (MD) with 95% confidence intervals (CI) of global quality of life (QOL), social and physical function scores, stratified by preoperative body mass index (BMI) <25 and ≥25. A Cox proportional hazards model produced hazard ratios (HR) with 95% CI for overall 5-year survival. Of 358 patients, 196 (55%) had preoperative BMI ≥25. Very severe and severe nutritional problems were associated with worse HRQOL in both BMI groups. E.g. MD's for global QOL among 'very severe' group was -29 (95% CI -39⁻-19) and -20 (95% CI -29⁻-11) for <25 and ≥25 BMI, respectively, compared to the 'low' group. Overall 5-year survival among 'very severe' and BMI ≥ 25 was worse; HR 4.6 (95% CI 1.4⁻15.6). Intense nutritional problems negatively impact postoperative HRQOL and combined with preoperative BMI ≥ 25 are associated with poorer 5-year overall survival representing a group needing greater clinical attention.

Influence of tube potential on CT body composition analysis.

OBJECTIVES: Our purpose was to investigate whether tube potential in contrast-enhanced computed tomography (CT) affects body composition analysis. METHODS: Images from dual-source, dual-energy CT from the abdomen with intravenous contrast media administration were used. A total of 17 patients (11 women, mean age 52) with a mean body mass index of 20.8 kg/cm2 were included. Simultaneously acquired images with a tube voltage of 80 kV and 140 kV were compared. Body composition was analyzed on a single slice at the L3 level. Parameters evaluated included muscle and fat attenuation (Hounsfield units [HU]), skeletal muscle index (cm2/m2), muscle area (cm2), and steatotic muscle area (cm2). Significant differences between 80 kV and 140 kV series were compared using the paired Student's t test. RESULTS: Tube potential affected muscle attenuation with an average difference of 17% between 80 kV and 140 kV series (48 HU versus 41 HU, P < 0.01), fat attenuation (-84 HU versus -69 HU, P < 0.01), skeletal muscle index of 5.2% (40.1 cm2/m2 versus 42.2 cm2/m2, P < 0.01), muscle area of 5.1% (117 cm2 versus 123 cm2, P < 0.01), and steatotic muscle area of 12.9% (31 cm2 versus 35 cm2, P < 0.01). CONCLUSION: Tube potential significantly affects body segmentation in contrast-enhanced CT.

Pancreatic Exocrine Insufficiency after Bariatric Surgery.

Morbid obesity is a lifelong disease, and all patients require complementary follow-up including nutritional surveillance by a multidisciplinary team after bariatric procedures. Pancreatic exocrine insufficiency (PEI) refers to an insufficient secretion of pancreatic enzymes and/or sodium bicarbonate. PEI is a known multifactorial complication after upper gastrointestinal surgery, and might constitute an important clinical problem due to the large number of bariatric surgical procedures in the world. Symptoms of PEI often overlap with sequelae of gastric bypass, making the diagnosis difficult. Steatorrhea, weight loss, maldigestion and malabsorption are pathognomonic for both clinical conditions. Altered anatomy after bypass surgery can make the diagnostic process even more difficult. Fecal elastase-1 (FE1) is a useful diagnostic test. PEI should be considered in all patients after bariatric surgery with prolonged gastrointestinal complaints that are suggestive of maldigestion and/or malabsorption. Appropriate pancreatic enzyme replacement therapy should be part of the treatment algorithm in patients with confirmed PEI or symptoms suggestive of this complication.

Preclinical evaluation of NF-κB-triggered dendritic cells expressing the viral oncogenic driver of Merkel cell carcinoma for therapeutic vaccination.

BACKGROUND: Merkel cell carcinoma (MCC) is a rare but very aggressive skin tumor that develops after integration of a truncated form of the large T-antigen (truncLT) of the Merkel cell polyomavirus (MCV) into the host's genome. Therapeutic vaccination with dendritic cells (DCs) loaded with tumor antigens is an active form of immunotherapy, which intends to direct the immune system towards tumors which express the respective vaccination antigens. METHODS: Cytokine-matured monocyte-derived DCs of healthy donors and MCC patients were electroporated with mRNA encoding the truncLT. To permit major histocompatibility complex (MHC) class II next to class I presentation, we used an RNA construct in which the antigen was fused to a DCLamp sequence in addition to the unmodified antigen. To further improve their immunogenicity, the DCs were additionally activated by co-transfection with the constitutively active nuclear factor (NF)-κB activator caIKK. These DCs were used to stimulate autologous CD8+ T-cells or a mixture of CD4+ and CD8+ T-cells. Then the percentage of T-cells, specific for the truncLT, was quantified by interferon (IFN)γ ELISpot assays. RESULTS: Both the truncLT and its DCLamp-fusion were detected within the DCs by flow cytometry, albeit the latter required blocking of the proteasome. The transfection with caIKK upregulated maturation markers and induced cytokine production. After 2-3 rounds of stimulation, the T-cells from 11 out of 13 healthy donors recognized the antigen. DCs without caIKK appeared in comparison less potent in inducing such responses. When using cells derived from MCC patients, we could induce responses for 3 out of 5 patients; however, here the caIKK-transfected DCs did not display their superiority. CONCLUSION: These results show that optimized DCs are able to induce MCV-antigen-specific T-cell responses. Therapeutic vaccination with such transfected DCs could direct the immune system against MCC.