Search

Background: Literature remains scarce on patients experiencing weight recurrence after initial adequate weight loss following primary bariatric surgery. Therefore, this study compared the extent of weight recurrence between patients who received a Sleeve Gastrectomy (SG) versus Roux-en-Y gastric bypass (RYGB) after adequate weight loss at 1-year follow-up. Methods: All patients undergoing primary RYGB or SG between 2015 and 2018 were selected from the Dutch Audit for Treatment of Obesity. Inclusion criteria were achieving ≥ 20% total weight loss (TWL) at 1-year and having at least one subsequent follow-up visit. The primary outcome was ≥ 10% weight recurrence (WR) at the last recorded follow-up between 2 and 5 years, after ≥ 20% TWL at 1-year follow-up. Secondary outcomes included remission of comorbidities at last recorded follow-up. A propensity score matched logistic regression analysis was used to estimate the difference between RYGB and SG. Results: A total of 19.762 patients were included, 14.982 RYGB and 4.780 SG patients. After matching 4.693 patients from each group, patients undergoing SG had a higher likelihood on WR up to 5-year follow-up compared with RYGB [OR 2.07, 95% CI (1.89–2.27), p< 0.01] and less often remission of type 2 diabetes [OR 0.69, 95% CI (0.56–0.86), p< 0.01], hypertension (HTN) [OR 0.75, 95% CI (0.65–0.87), p< 0.01], dyslipidemia [OR 0.44, 95% CI (0.36–0.54), p< 0.01], gastroesophageal reflux [OR 0.25 95% CI (0.18–0.34), p< 0.01], and obstructive sleep apnea syndrome (OSAS) [OR 0.66, 95% CI (0.54–0.8), p< 0.01]. In subgroup analyses, patients who experienced WR after SG but maintained ≥ 20%TWL from starting weight, more often achieved HTN (44.7% vs 29.4%), dyslipidemia (38.3% vs 19.3%), and OSAS (54% vs 20.3%) remission compared with patients not maintaining ≥ 20%TWL. No such differences in comorbidity remission were found within RYGB patients. Conclusion: Patients undergoing SG are more likely to experience weight recurrence, and less likely to achieve comorbidity remission than patients undergoing RYGB.

In recent years, the contingency-based management accounting literature has been criticized for being fragmentary and contradictory as a result of methodological limitations. This study adds to this picture by showing that the theoretical meaning of some commonly used statistical techniques is unclear, i.e. the functional forms are not precise enough to be able to discriminate between several sometimes even conflicting theories of contingency fit. The study also shows that the techniques differ significantly in terms of how interaction effects between context and management accounting are modeled. This implies that some methods are only appropriate when theory predicts interaction effects in general while others are only appropriate in cases where theory specifies a more precise functional form of interaction such as symmetrical or crossover interactions. Based on these observations, several recommendations for future research are proposed.

In recent years factors that may influence firms' decisions to adopt target costing have been presented in the literature. These studies generally argue that target costing adoption rates positively correlate with the intensity of competition and perceived environmental uncertainty. However, empirical support for this view is limited. This study argues that the adoption of target costing positively correlates with the intensity of competition, but negatively correlates with perceived environmental uncertainty. In addition, perceived environmental uncertainty moderates the adoption of target costing when the intensity of competition intensifies. Hypotheses are generated and tested using data provided by a web-based survey of Swedish manufacturing firms. The findings indicate that the adoption of target costing and the intensity of competition positively relate, although the effect reduces with an increase in perceived environmental uncertainty. There is no evidence of a direct relationship between perceived environmental uncertainty and the adoption of target costing. As a result, the findings partly conflict with the outcomes of earlier work. The article also discusses the implications for research and firm managers considering the adoption of target costing. Keywords: Target costing Adoption Perceived environmental uncertainty Intensity of competition

Contingency literature in the accounting control area has for a long time been criticized for being fragmentary and contradictory as a result of methodological limitations. A review of 10 articles in the strategy-MAS area adds to this picture by showing that many different forms of fit have been used, and that very few researchers fully acknowledge the difficulties of relating these forms to each other. As a result, some researchers claim that their findings are contradictory when this is not necessarily the case, while others incorrectly argue that their results are strongly supported by previous studies.

The effect of surgically induced weight loss on the quality of life of morbidly obese patients was investigated in a controlled cross-sectional study. A postoperative group of 62 patients was compared with a control group of 20 preoperative patients. The Nottingham Health Profile part I (NHP-I) and part II (NHP-II) and a visual analogue scale were used for quality of life assessment. Significant differences were found on the NHP-II (P [is less than] 0.0001), the visual analogue scale (P [is less than] 0.001), and on the domains of mobility (P [is less than] 0.0001), energy (P [is less than] 0.001), and emotional reaction (P [is less than] 0.001) on the NHP-I in favor of the post-operative group. Better quality of life outcome was related to larger weight loss and shorter length of postoperative follow-up. No correlation was found between quality of life outcome and the type of surgical procedure or surgical complications. The unemployment rate was 53% for the overall postoperative group and 64% for the female postoperative group compared with 80% (P [is less than] 0.05) and 84% (NS) in the overall and female control groups, respectively. Overweight as the reason for unemployment was more frequent in the control group. The results of this study show that quality of life is better after surgically induced weight loss and is not related to the type of surgical procedure nor to surgical complications. Postoperative quality of life tends to decrease with time.

Non-alcoholic fatty liver disease (NAFLD) is a widespread liver disease in Western society, but its multifactorial pathogenesis is not yet fully understood. Ultrastructural analysis of liver sinusoidal endothelial cells (LSECs) in animal models and in vitrostudies shows defenestration early in the course of NAFLD, promoting steatosis. LSECs and fenestrae are important in the transport of lipids across the sinusoids. However, human ultrastructural data, especially on LSECs and fenestrae, are scarce. This study aimed to explore the ultrastructural changes in perfusion type fixed liver biopsies of NAFLD patients with and without non-alcoholic steatohepatitis (NASH), with a special focus on LSECs and their fenestration. Liver biopsies from patients with NAFLD were fixed using two perfusion techniques, jet and injection fixation, for needle and wedge biopsies, respectively. Ultrastructural changes were studied using transmission electron microscopy. NASH was diagnosed by bright-field microscopy using the SAF score (steatosis, activity, fibrosis). Thirty-seven patients were included, of which 12 (32.4%) had NASH. Significantly less defenestration was found in NASH compared to noNASH samples (p=0.002). Other features, i.e., giant mitochondria and fenestrae size did not differ between groups. Furthermore, we described new structures, i.e., single cell steatonecrosis and inflammatory fat follicles (IFF) that were observed in both groups. Concluding, defenestration was more common in noNASH compared to NASH in human liver samples. Defenestration was not related to the degree of steatosis or fibrosis. We speculate that defenestration can be a protective mechanism in simple steatosis which is lacking in NASH.

Background: Current studies mainly focus on total weight loss and comorbidity reduction. Only a few studies compare Quality of Life (QoL) after sleeve gastrectomy (SG) and Roux-en-Y gastric bypass (RYGB). This study was conducted to examine the extent of improvement in QoL on different domains after primary bariatric surgery and compare these results to Dutch reference values. Methods: The study included prospectively collected data from patients who underwent primary bariatric surgery in five Dutch hospitals. The RAND-36 questionnaire was used to measure the patient’s QoL; preoperatively and twelve months postoperatively. Postoperative scores were compared to Dutch reference values, standardized for age, using t-test. A difference of more than 5% was considered a minimal important difference. A multivariate linear regression analysis was used to compare SG and RYGB on the extent of improvement, adjusted for case-mix factors. Results: In total, 4864 patients completed both the pre- and postoperative questionnaire. Compared with Dutch reference values, patients postoperatively reported clinically relevant better physical functioning(RYGB + 6.8%), physical role limitations(SG + 5.6%; RYGB + 6.2%) and health change(SG + 77.1%; RYGB + 80.0%), but worse general health perception(SG − 22.8%; RYGB − 17.0%). Improvement in QoL was similar between SG and RYGB, except for physical functioning(β2.758; p-value 0.008) and general health perception(β2.607; p-value < 0.001) for which RYGB patients improved more. Conclusions: SG and RYGB patients achieved a better postoperative score in physical functioning, physical role limitationsand health change comparedto Dutch reference values, and a worse score in general health perception.

Background: Revisional bariatric surgery (RBS) constitutes a possible solution for patients who experience an inadequate response following bariatric surgery or significant weight regain following an initial satisfactory response. This paper reports results from the first modified Delphi consensus-building exercise on RBS. Methods: We created a committee of 22 recognised opinion-makers with a special interest in RBS. The committee invited 70 RBS experts from 27 countries to vote on 39 statements concerning RBS. An agreement amongst ≥ 70.0% experts was regarded as a consensus. Results: Seventy experts from twenty-seven countries took part. There was a consensus that the decision for RBS should be individualised (100.0%) and multi-disciplinary (92.8%). Experts recommended a preoperative nutritional (95.7%) and psychological evaluation (85.7%), endoscopy (97.1%), and a contrast series (94.3%). Experts agreed that Roux-Y gastric bypass (RYGB) (94.3%), One anastomosis gastric bypass (OAGB) (82.8%), and single anastomosis duodeno-ileal bypass with sleeve gastrectomy (SADI-S) (71.4%) were acceptable RBS options after gastric banding (84.3%). OAGB (84.3%), bilio-pancreatic diversion/duodenal switch (BPD/DS) (81.4%), and SADI-S (88.5%) were agreed as consensus RBS options after sleeve gastrectomy. lengthening of bilio-pancreatic limb was the only consensus RBS option after RYGB (94.3%) and OAGB (72.8%). Conclusion: Experts achieved consensus on a number of aspects of RBS. Though expert opinion can only be regarded as low-quality evidence, the findings of this exercise should help improve the outcomes of RBS while we develop robust evidence to inform future practice.

To the Editor: In the study by Alfa and colleagues (February 1999),[1] the effects of maximizing oxygen delivery in patients with severe sepsis and septic shock were studied. Although reduction [...]

Nonalcoholic fatty liver disease (NAFLD) comprises fat-accumulating conditions within hepatocytes that can cause severe liver damage and metabolic comorbidities. Studies suggest that mitochondrial dysfunction contributes to its development and progression and that the hepatic lipidome changes extensively in obesity and in NAFLD. To gain insight into the relationship between lipid metabolism and disease progression through different stages of NAFLD, we performed lipidomic analysis of plasma and liver biopsy samples from obese patients with nonalcoholic fatty liver (NAFL) or nonalcoholic steatohepatitis (NASH) and from those without NAFLD. Congruent with earlier studies, hepatic lipid levels overall increased with NAFLD. Lipid species that differed with NAFLD severity were related to mitochondrial dysfunction; specifically, hepatic cardiolipin and ubiquinone accumulated in NAFL, and levels of acylcarnitine increased with NASH. We propose that increased levels of cardiolipin and ubiquinone may help to preserve mitochondrial function in early NAFLD, but that mitochondrial function eventually fails with progression to NASH, leading to increased acylcarnitine. We also found a negative association between hepatic odd-chain phosphatidylcholine and NAFLD, which may result from mitochondrial dysfunction-related impairment of branched-chain amino acid catabolism. Overall, these data suggest a close link between accumulation of specific hepatic lipid species, mitochondrial dysfunction, and the progression of NAFLD.

ObjectiveObesity is a risk factor for non-alcoholic steatohepatitis (NASH). This risk has been attributed to visceral adipose tissue (vAT) expansion associated with increased proinflammatory mediators. Accumulation of CD11c+proinflammatory adipose tissue macrophages (ATM) is an important driver of vAT inflammation. We investigated the role of ATMs in hepatic inflammation during NASH development.DesignvAT isolated from lean, obese or ATM-depleted (using clodronate liposomes) obese mice was transplanted to lean ldlr-/-acceptor mice. Systemic and hepatic inflammation was assessed either after 2 weeks on standard chow or after 8 weeks on high cholesterol diet (HCD) to induce NASH.ResultsTransplanting donor vAT from obese mice increased HCD-induced hepatic macrophage content compared with lean-transplanted mice, worsening liver damage. ATM depletion prior to vAT transplantation reduced this increased hepatic macrophage accumulation. On chow, vAT transplantation induced a more pronounced increase in circulating and hepatic neutrophil numbers in obese-transplanted than lean-transplanted mice, while ATM depletion prior to vAT transplantation reversed this effect. Microarray analysis of fluorescence-activated cell sorting of CD11c+and CD11c−macrophages isolated from donor adipose tissue showed that obesity resulted in enhanced expression of neutrophil chemotaxis genes specifically in CD11c+ATMs. Involvement of the neutrophil chemotaxis proteins, CXCL14 and CXCL16, was confirmed by culturing vAT. In humans, CD11c expression in vAT of obese individuals correlated with vAT expression of neutrophil chemotactic genes and with hepatic expression of neutrophil and macrophage marker genes.ConclusionATMs from obese vAT induce hepatic macrophage accumulation during NASH development, possibly by enhancing neutrophil recruitment.

Exclusion of the proximal small intestine with the novel nonsurgical duodenal-jejunal bypass liner in combination with dietary intervention induces superior weight loss and rapid improvement of glycemic control in obese patients with type 2 diabetes in comparison with dietary treatment alone.

Incisional hernias in old stoma wounds occur in one-third of former stoma patients and pose a significant clinical problem. Parastomal hernias can be prevented by prophylactic mesh placement; however, no trial results are available for incisional hernia prevention after stoma reversal. In this feasibility study, we explore the safety of placing an intraperitoneal mesh to prevent incisional herniation after temporary stoma reversal. Ten patients who underwent a low anterior resection with a deviating double-loop stoma for rectal cancer received an intraperitoneal parastomal mesh at the time of stoma formation. At stoma reversal, laparoscopy was performed and adhesions were scored. After reversal, the mesh defect was closed. Mesh and stoma complications were closely monitored. Incisional herniation was assessed at the 2-year follow-up after stoma reversal using ultrasonography. No infections occurred after mesh placement. After a median of 6 months, stomas were reversed. Laparoscopy could be performed in seven patients; all patients had adhesions (median of 25 % of mesh surface). In three patients, the bowel was involved; one required a laparotomy for bowel mobilization during stoma reversal. No adhesion-related morbidity was noted at any time. Except for one superficial wound infection after stoma reversal, no infectious complications were observed. After a median follow-up of 26 months, no incisional herniations were demonstrated. Prophylactic mesh placement in temporary stoma formations seems safe and feasible and prevents incisional herniation 2 years after stoma reversal.

Background:Mesenchymal stem cells (MSCs) offer promise as therapeutic aids in the repair of tendon and ligament disorders in sport horses. Equine allogeneic MSCs derived from umbilical cord blood (eUCB-MSCs) can be obtained in a minimally invasive fashion with successful propagation of MSCs.Objective:The objective of this study was to determine the applicability and therapeutic effect of eUCB-MSCs on tendinitis of the superficial digital flexor tendon, desmitis of the suspensory ligament, tendinitis of the deep digital flexor tendon, and desmitis of the inferior check ligament in clinical cases.Methods:A retrospective clinical study was performed. At two equine clinics, 52 warmblood horses were treated with cultured eUCB-MSCs between 2009 and 2012. About 2–10 × 106cells per lesion were administered. When a lesion was treated twice, the total amount could run up to 20 × 106cells. Pearson's chi-squared test was used to compare the effect of the injured structure on the success rate, as well as the effect of the age of the horse.Results:Based on repeated examinations, 40 horses (77%) returned to work on the same or a higher level based on information provided by the owner. Neither the injured structure nor the age of the horse had a statistically significant influence on the result.Conclusion:Overall, the results of treatment of some tendon and ligament injuries with eUCB-MSCs in clinical cases are promising.

The development of an immunosuppressive state during the protracted course of sepsis is associated with opportunistic infections and is considered to correlate with the extent of the proinflammatory response during early sepsis. Short-term intervention with enteral lipid-rich nutrition was shown to attenuate the acute inflammatory response. This study investigates the effects of lipid-rich nutrition on the immunosuppression induced by polymicrobial sepsis. Female BALBc mice were either fasted or fed liquid lipid-rich nutrition or isocaloric control nutrition before and shortly after induction of polymicrobial sepsis through cecal ligation and puncture (CLP) or sham operation. After 4 days, mice were intranasally infected with Pseudomonas aeruginosa. Twenty-four hours after P. aeruginosainfection, fasted and control nutrition-fed CLP mice displayed a significantly higher bacterial load in the lungs than did corresponding sham-operated mice (P< 0.001 and P< 0.05, respectively). Fasted CLP mice expressed reduced pulmonary levels of proinflammatory cytokines interleukin 12 (IL-12) and interferon (IFN-) in comparison to sham mice (both P< 0.05). Lipid-rich nutrition prevented the increase in bacteria, promoted the IL-12 and IFN- production (IL-12 and IFN- P< 0.05 vs. fasted and IFN- P< 0.05 vs. control nutrition), and prevented the expression of the immunosuppressive cytokine IL-10 (P< 0.05 vs. control nutrition) in lungs of CLP mice. The preserved immune defense during late sepsis in lipid-rich fed mice was preceded by attenuation of the early inflammatory response (IL-6 P= 0.05 and IL-10 P< 0.01 vs. fasted CLP mice) at 6 h after CLP. In conclusion, short-term treatment with lipid-rich enteral nutrition improves the pulmonary antimicrobial defense during polymicrobial sepsis.

Laparoscopic incisional hernia repair with intraperitoneal mesh is associated with a certain degree of adhesion formation to the mesh. This experimental study examined the efficacy of several coated meshes for adhesion reduction.Five commercially available meshes with a layered coating were placed intraperitoneally in rats and followed up for 90 days: polypropylene and polyester meshes, both coated with absorbable collagen (Parietene Composite and Parietex Composite, respectively), and three polypropylene meshes respectively coated with absorbable omega-3 fatty acids (C-Qur Edge), absorbable cellulose (Sepramesh IP), and nonabsorbable expanded polytetrafluoroethylene (Intramesh T1). Uncoated polypropylene and collagen meshs (Parietene and Permacol, respectively) served as the control condition. Adhesions, incorporation, and tissue reaction were evaluated macro- and microscopically. Additionally, the development of the neoperitoneum was examined.All the coated meshes performed equally well in terms of adhesion reduction. The collagen mesh performed comparably, but the uncoated polypropylene mesh performed significantly worse. The different coatings led to very differing degrees of inflammation. Ingrowth was observed only at the place of suture but was comparable for all the meshes except C-Qur Edge, which showed the weakest incorporation. Development of a neoperitoneum on the mesh surface occurred independently of whether an absorbable or nonabsorbable coating or no coating at all was present.Commercially available meshes with a layered coating deliver comparable adhesion reduction. The physical presence of a layered coating between the intraperitoneal content and the abdominal wall seems to be more important than the chemical properties of the coating in adhesion formation.

Chronic inflammation is characterized by the influx of neutrophils and is associated with an increased production of reactive oxygen species that can damage DNA. Oxidative DNA damage is generally thought to be involved in the increased risk of cancer in inflamed tissues. We previously demonstrated that activated neutrophil mediated oxidative stress results in a reduction in nucleotide excision repair (NER) capacity, which could further enhance mutagenesis. Inflammation and oxidative stress are critical factors in the progression of nonalcoholic fatty liver disease that is linked with enhanced liver cancer risk. In this report, we therefore evaluated the role of neutrophils and the associated oxidative stress in damage recognition and DNA repair in steatotic livers of 35 severely obese subjects with either nonalcoholic steatohepatitis (NASH) (n=17) or steatosis alone (n=18). The neutrophilic influx in liver was assessed by myeloperoxidase (MPO) staining and the amount of oxidative DNA damage by measuring M1dG adducts. No differences in M1dG adduct levels were observed between patients with or without NASH and also not between individuals with high or low MPO immunoreactivity. However, we found that high expression of MPO in the liver, irrespective of disease status, reduced the damage recognition capacity as determined by staining for histone 2AX phosphorylation (γH2AX). This reduction in γH2AX formation in individuals with high MPO immunoreactivity was paralleled by a significant decrease in NER capacity as assessed by a functional repair assay, and was not related to cell proliferation. Thus, the observed reduction in NER capacity upon hepatic inflammation is associated with and may be a consequence of reduced damage recognition. These findings suggest a novel mechanism of liver cancer development in patients with nonalcoholic fatty liver disease.

Endotoxin is hypothesized to play an important role in the activation of inflammatory pathways associated with nonalcoholic steatohepatitis (NASH). However, demonstration of hepatic endotoxin exposure is challenging due to the inaccessibility of the portal circulation. Furthermore, reliable measurement of the relatively low endotoxin levels in plasma of patients with liver disease and subsequent interpretation remain difficult.

Background:Affordable, easytouse, and reliable CD4T lymphocyte enumeration systems are needed in resourceconstrained settings to monitor HIV.Methods:A simple image cytometer was used to count fluorescently labeled CD4T and CD8T lymphocytes from CD3 immunomagnetically selected cells on blood specimens of 460 HIV1infected patients in Siriraj Hospital, Bangkok, Thailand. Results were compared with flow cytometry FCM.Results:CD4T lymphocyte counts by image cytometer were comparable R≥ 0.97 with those by the FACSCount and the FACScan with a bias of 7.3 and 9.1, respectively. At very low CD4T lymphocyte counts ≤50μl some overcount outliers were observed by the FACScan and image cytometer when compared with the FACSCount. For CD8 enumeration, the image cytometer showed a good correlation R 0.96 and a consistent undercount ∼17 when compared with the FACSCount.Conclusions:Evaluation of the image cytometer for CD4 and CD8 enumeration demonstrated comparable results with FCM on a population of HIV1infected patients. The image cytometer is a good alternative method for pointofcare settings in resourceconstrained countries. © 2009 Clinical Cytometry Society

Inflammation and oxidative stress are considered critical factors in the progression of nonalcoholic fatty liver disease. Myeloperoxidase (MPO) is an important neutrophil enzyme that can generate aggressive oxidants; therefore, we studied the association between MPO and nonalcoholic fatty liver disease. The distribution of inflammatory cells containing MPO in liver biopsies of 40 severely obese subjects with either nonalcoholic steatohepatitis (NASH) (n= 22) or simple steatosis (n= 18) was investigated by immunohistochemistry. MPO-derived oxidative protein modifications were identified by immunohistochemistry and correlated to hepatic gene expression of CXC chemokines and M1/M2 macrophage markers as determined by quantitative PCR. MPO plasma levels were determined by ELISA. The number of hepatic neutrophils and MPO-positive Kupffer cells was increased in NASH and was accompanied by accumulation of hypochlorite-modified and nitrated proteins, which can be generated by the MPO-H2O2system. Liver CXC chemokine expression was higher in patients with accumulation of MPO-mediated oxidation products and correlated with hepatic neutrophil sequestration. Plasma MPO levels were elevated in NASH patients. Interestingly, neutrophils frequently surrounded steatotic hepatocytes, resembling the crown-like structures found in obese adipose tissue. Furthermore, hepatic M2 macrophage marker gene expression was increased in NASH. Our data indicate that accumulation of MPO-mediated oxidation products, partly derived from Kupffer cell MPO, is associated with induction of CXC chemokines and hepatic neutrophil infiltration and may contribute to the development of NASH.

Background:We developed a volumetric single platform image cytometer SP ICM that is dedicated to count CD4and CD8T lymphocytes for HIV monitoring in resourceconstrained settings. The instrument was designed to be lowcost, yet reliable, easytouse, and robust.Methods:Whole blood is incubated with CD3magnetic nanoparticles, CD4phycoerythrin PE, and CD8peridininchlorophyllprotein complex PerCP. The CD3 cells are immunomagnetically attracted to an analysis surface, where fluorescence images of CD4and CD8T lymphocytes are recorded and analyzed, respectively. We compared CD4, CD8 counts, and CD4CD8 ratio obtained by the SP ICM with those from a SP flow cytometer FCM tetraCXP method on blood samples from 145 patients.Results:Good correlations were obtained R: 0.96–0.99 between the SP ICM and the SP FCM. There was ∼10 CD8 undercount in the SP ICM, which could be partly caused by CD8dimT lymphocytes that were not detected by the instrument or not counted by the image analysis due to the crosstalk from the CD4PE signal in the CD8PerCP image.Conclusions:The SP ICM is a good candidate for HIV monitoring in pointofcare settings of resourceconstrained countries. © 2008 Clinical Cytometry Society

HIV monitoring in resource‐constrained settings demands affordable and reliable CD4+ T lymphocytes enumeration methods. We developed a simple single platform image cytometer (SP ICM), which is a dedicated volumetric CD4+ T lymphocytes enumeration system that uses immunomagnetic and immunofluorescent technologies. The instrument was designed to be a low‐cost, yet reliable and robust one. In this article we test the instrument and the immunochemical procedures used on blood from HIV negative and HIV positive patients.After CD4 immunomagnetic labeling in whole blood, CD4+ T lymphocytes, CD4+dim monocytes and some nonspecifically labeled cells are magnetically attracted to an analysis surface. Combining with CD3‐Phycoerythrin (PE) labeling, only CD3+CD4+ T lymphocytes are fluorescently labeled and visible in a fluorescent image of the analysis surface. The number of CD4+ T lymphocytes is obtained by image analysis. Alternatively, CD3 immunomagnetic selection in combination with CD4 immunofluorescent labeling can also be applied for CD4+ T lymphocytes enumeration.The SP ICM system was compared with two single platform flow cytometer (SP FCM) methods: tetraCXP and TruCount methods. The SP ICM system has excellent precision, accuracy and linearity for CD4+ T lymphocytes enumeration. Good correlations were obtained between the SP ICM and the SP FCM methods for blood specimens of 44 HIV− patients, and of 63 HIV+ patients. Bland–Altman plots showed interchangeability between the SP ICM and the SP FCM methods.The immunolabeling methods and the instrumentation are simple and easy‐to‐handle for less‐trained operators. The SP ICM system is a good candidate for CD4+ T lymphocytes enumeration in point‐of‐care settings of resource‐constrained countries. © 2007 Clinical Cytometry Society.

In clinical research, increased permeability has been scrutinized as a potential indicator of the severity of gastrointestinal disease and as a potential cause of the perpetuation of severe inflammatory activity in infectious states. This review discusses old and recent epidemiological and clinical evidence to establish whether increased permeability in sepsis is a sequel or a cause of multiple organ failure. In addition, old and new evidence linking inflammation and permeability in abnormal gastrointestinal anatomy and function to liver abnormalities in susceptible patients will be reviewed.

ABSTRACTSimplification of cell enumeration technologies is necessary, especially for resource-poor countries, where reliable and affordable enumeration systems are greatly needed. In this paper, an immunomagnetic single-platform image cytometer (SP ICM) for cell enumeration based on antibody specificity is reported. A chamber/magnet assembly was designed such that the immunomagnetically labeled, acridine orange-stained cells in a blood sample moved to the surface of the chamber, where a fluorescent image was captured and analyzed for cell enumeration. The system was evaluated by applying one kind of antibody to count leukocytes and one kind for each leukocyte subpopulation: CD45 for leukocytes, CD3 for T lymphocytes, and CD19 for B lymphocytes. Excellent precision and linearity were achieved. Moreover, these cell counts, each from blood specimens of 42 to 52 randomly selected patients, were compared with those obtained by SP (TruCount) and dual-platform (DP) flow cytometry (FCM) technologies. The cell counts obtained by our system were in between those obtained from the TruCount and DP FCM methods; and good correlations were achieved (R= 0.95). For CD4+counts, as we expected, the cell count by our system was significantly higher than the CD4+T-lymphocyte counts obtained by SP and DP FCM methods. Immunophenotyping of the immunomagnetically selected CD4+cells showed that, besides CD4+T lymphocytes, a proportion of the CD4+dim monocytes was also selected. Our system is a simple immunomagnetic SP ICM, which can potentially be used for enumeration of CD3+CD4+T lymphocytes in resource-poor countries if an additional CD3 immunofluorescent label is applied.

For resource‐poor countries, affordable methods are required for enumeration of CD4+ T lymphocytes of HIV‐positive patients. For infants, additional determination of CD4/CD8 ratio is needed.We determine the CD4+ and CD8+ T lymphocytes as the CD3+CD4+ and CD3+CD8+ population of blood cells. Target cells are CD3‐immunomagnetically separated from the whole blood, and CD4‐Phycoerythrin and CD8‐PerCP immunofluorescently labeled. A point‐of‐care single platform image cytometer was developed to enumerate the target CD3+CD4+ and CD3+CD8+ populations. It has light‐emitting diodes illumination, is fully computer‐controlled, operates from a 12 V battery, and was designed to be cheap and easy‐to‐handle. Target cells are imaged on a CCD camera and enumerated by an image analysis algorithm. The cytometer outputs the absolute number of CD4+ and CD8+ T lymphocytes/μl and CD4/CD8 ratio.The quality of the cell images obtained with the cytometer is sufficient for a reliable enumeration of target cells. The image cytometer achieves an accuracy of better than 10% in the range of 50–1700 cells/μl. Analysis of blood samples from HIV patients yields a good agreement with the TruCount method for CD4 and CD8 count and CD4/CD8 ratio.The image cytometer is affordable (component costs $3,000), compact (25 × 25 × 20 cm3), and uses disposable test materials, making it a good candidate to monitor progression of immunodeficiency disease in resource‐poor settings. © 2007 International Society for Analytical Cytology.

Morbid obesity is associated with serious co-morbidities as cardiovascular disease, insulin resistance (IR) and type 2 diabetes (T2DM). Conventional therapy for T2DM consists of weight loss and pharmacotherapy. Recently, surgery was suggested to be the best treatment for T2DM in morbid obese patients. This review will focus on the effects of bariatric surgery on T2DM. The outcomes of different bariatric techniques will be discussed. Moreover, the question why weight loss is advantageous in treating T2DM will be addressed. In this view, the effect of decreased nutrient intake, decreased intracellular fatty acid accumulation as well as decreased body fat mass on T2DM are discussed.

Laparoscopic Nissen fundoplication offers clear benefits for our patients, but requires advanced laparoscopic skills. Robotic assistance in laparoscopic antireflux surgery improves dexterity skills and shortens learning curve, possibly leading to faster, more precise, and safer laparoscopic surgery.

Background: This study was designed as an economic evaluation alongside a randomized clinical trial. The object of this study was to evaluate the 1-year cost-effectiveness of surgical treatment of morbid obesity comparing two operations. Methods: 100 patients were assigned randomly to vertical banded gastroplasty (VBG) or Lap-Band surgery. Both medical and non-medical costs were identified and measured. Costs data were combined with percentage Excess Weight Loss (%EWL) and with Quality Adjusted Life Years (QALYs) to obtain cost per %EWL and cost per QALY ratios. Results: At 1 year, the total costs were not significantly different between both groups (95% confidence interval E5,999 - E1,765). Also, the QALY gain after surgery was not significantly different between the two groups. However, %EWL was significantly higher in the VBG group compared to the Lap-Band group, P-value .0001. The estimated incremental cost per %EWL was E105.83 (E1,885.91/-17.82). For the costs per QALY, the estimated ratio was dominant. The overall mortality in this study was 2%. 2 patients in the VBG group died within 30 days after surgery; 1 of these deaths was possibly related to the VBG procedure. Conclusion: At 1 year after surgery, the costs and QoL of the two treatment modalities were found to be equal. Therefore, the selection of the procedure can be based on the clinical aspects, effectivity and safety at 1 year. In addition, the results of a long-term cost-effectiveness analysis (e.g. with a follow-up of 36 months) planned in the future can also be helpful in the selection of the preferred treatment.

Introduction To review systematically the effect of interventions aimed at hemodynamic optimization and to relate this to the quality of individual published trials.Methods A systematic, computerized bibliographic search of published studies and citation reviews of relevant studies was performed. All randomized clinical trials in which adult patients were included in a trial deliberately aiming at an optimized or maximized hemodynamic condition of the patients (with oxygen delivery, cardiac index, oxygen consumption, mixed venous oxygen saturation and/or stroke volume as end-points) were selected. A total of 30 studies were selected for independent review. Two reviewers extracted data on population, intervention, outcome and methodological quality. Agreement between reviewers was high: differences were eventually resolved by third-party decision. The methodological quality of the studies was moderate (mean 9.0, SD 1.7), and the outcomes of the randomized clinical trials were not related to their quality.Results Efforts to achieve an optimized hemodynamic condition resulted in a decreased mortality rate (relative risk ratio (RR) 0.75 (95% confidence interval (CI) 0.62 to 0.90) in all studies combined. This was due to a significantly decreased mortality in peri-operative intervention studies (RR 0.66 (95% CI 0.54 to 0.81). Overall, patients with sepsis and overt organ failure do not benefit from this method (RR 0.92 (95% CI 0.75 to 1.11)).Conclusion This systematic review showed that interventions aimed at hemodynamic optimization reduced mortality. In particular, trials including peri-operative interventions aimed at the hemodynamic optimization of high-risk surgical patients reduce mortality. Overall, this effect was not related to the trial quality.

We demonstrate that in a sensor based on a multichannel Young interferometer, the phase information obtained for different pairs of channels can be used to correct the long-term instability (drift) due to temperature differences between measuring and reference channels, the drift in the alignment of the setup, etc. Experiments show that the nature of a major part of the drift is such that the drift present in one of the channels can be determined by interpolation of the drift measured in the two adjacent channels. It is shown that a drift reduction of 10 times can be achieved as compared with the situation in which no correction is applied. We anticipate that these findings will permit the exploitation of the extreme sensitivity of interference-based sensors to a much greater extent.

Probiotic therapy modulates the composition of the intestinal flora and inhibits the inflammatory response. These properties may be of benefit in the preservation of gut barrier integrity after injury or stress. In this study, we examined the effect of two Lactobacillus strains selected for their pathogen exclusion properties on intestinal barrier integrity following hemorrhagic shock. Additionally, the responsiveness of the macrophage cell line RAW 264.7 to combined exposure to Lactobacillus DNA or oligodeoxynucleotides containing CpG motifs (CpG-ODN) and endotoxin was assessed by measuring tumor necrosis factor alpha (TNF-α) release. Rats were administered lactobacilli (5 x 109 CFU) or vehicle for 7 days and were subjected subsequently to hemorrhagic shock by withdrawal of 2.1 ml blood/100 g tissue. Levels of plasma endotoxin, bacterial translocation to distant organs, and filamentous actin (F-actin) in the ileum were determined 24 h later. Rats treated with Lactobacillus rhamnosus showed reduced levels of plasma endotoxin (8 ± 2 pg/ml versus 24 ± 4 pg/ml; P = 0.01), bacterial translocation (2 CFU/gram versus 369 CFU/gram; P < 0.01), and disruption of F-actin distribution following hemorrhagic shock compared with nontreated control rats. In contrast, pretreatment with Lactobacillus fermentum had no substantial effect on gut barrier integrity. Interestingly, DNA preparations from both lactobacilli reduced endotoxin-induced TNF-α release dose dependently, whereas CpG-ODN increased TNF-α release. In conclusion, the pathogen exclusion properties of both Lactobacillus strains and the reduction of endotoxin-induced inflammation by their DNA in vitro are not prerequisites for a beneficial effect of probiotic therapy on gut barrier function following hemorrhagic shock. Although pretreatment with Lactobacillus spp. may be useful to preserve gut barrier integrity following severe hypotension, a thorough assessment of specific strains seems to be essential.

The structure of individual nucleosomes organized within reconstituted 208-12 arrays at different levels of compaction was examined by tapping mode atomic force microscopy in air and liquid. Reconstitution at lower histone octamer to DNA weight ratios showed an extended beads-on-a-string morphology with less than the expected maximum of 12 nucleosome core particles per array, each particle located in the most favored positioning site. A correlation of the contour lengths of these arrays with the number of observed particles revealed two distinct populations of particles, one with ∼50nm of bound DNA and a second population with ∼25nm. The measured nucleosome center-to-center distances indicate that this ∼25nm is not necessarily symmetrically bound about the dyad axis, but can also correspond to DNA bound from either the entry or exit point of the particle to a location at or close to the dyad axis. An assessment of particle heights suggests that particles wrapping ∼25nm of DNA are most likely to be subnucleosomal particles, which lack either one or both H2A-H2B dimers. At a higher reconstitution ratio, folded compact arrays fully populated with 12 nucleosome core particles, were observed. Liquid measurements demonstrated dynamic movements of DNA loops protruding from these folded arrays.

The new copper(I) coordination polymer poly[tri-μ2-cyano-C,N)-bis(μ2-2,3-dimethyl-pyrazine- N,N)] tricopper(I) (I) was prepared by the reaction of copper(I) cyanide with 2,3-dimethylpyrazine in acetonitrile. In the crystal structure of Ia novel CuCN substructure is found which is connected by the dimethylpyrazine ligands into a three-dimensional coordination network. The thermal properties of Iwere investigated using simultaneous differential thermoanalysis (DTA), thermogravimetry (TG) and mass spectrometry (MS) as well as temperature resolved X-ray powder diffraction. On heating, compound Ilooses a part of the dimethylpyrazine ligands in an endothermic reaction to form the known ligand poor compound (CuCN)2-(2,3-dimethylpyrazine) IIas an intermediate which decomposes to CuCN on further heating.

Intermittent clamping of the porta hepatis (PHC) is commonly performed during liver surgery to reduce blood loss and has been reported to precondition livers resulting in improved outcome after liver surgery (humans) and transplantation (animals). This study investigated the early expression of cytoprotective stress proteins during ischemia-reperfusion induced by PHC. Liver samples were taken before and after each event in a two-cycle ischemia-reperfusion protocol using 15 minutes of PHC followed by 5 minutes of reperfusion. Liver tissue was analyzed by real-time polymerase chain reaction for heme oxygenase (HO)-1 and heat shock protein (HSP)-70 mRNA expression. Extracted protein was analyzed by Western blot for HO-1, and HSP-70 and nuclear extracts were analyzed by DNA mobility shift assay for hypoxia inducible factor (HIF)-1 and heat shock factor (HSF)-1. Within minutes of PHC, significant increases in HO-1 mRNA expression were detected, and these were maintained throughout the protocol (P<0.01). Protein expression of HO-1 (P<0.03) and HO-1 activity (P<0.05) were similarly increased between the start and end of ischemia- reperfusion (40 minutes). Binding of active HIF-1 to its consensus sequence was increased within 15 minutes of the start of the ischemia-reperfusion cycle. Although evidence of the transcriptionally active form of HSF-1 was detected at the same time point, this was not reflected in measurable changes in HSP-70 mRNA or protein. In conclusion, expression of the cytoprotective protein HO-1 is significantly up-regulated in the liver within minutes of PHC. It is likely that HO-1 contributes to the early protective effects of ischemic preconditioning.

Background: Ghrelin is a recently discovered orexigenic gastric hormone, whose production is induced by lack of food in the stomach. In morbidly obese individuals, ghrelin levels are low compared to lean persons. During dieting, plasma ghrelin levels increase, leading to an orexigenic signal, which could explain the lack of success of dieting in morbidly obese individuals. Morbid obesity is best treated with bariatric surgery, in which gastric bypass is reported to be more effective than restrictive surgery. A possible explanation could be the difference in plasma ghrelin levels after both operations for bariatric surgery. In this study, plasma ghrelin levels were investigated during a 2-year follow-up. Methods: 17 morbidly obese patients received gastric restrictive surgery. Plasma ghrelin, leptin and insulin levels were evaluated preoperatively and 1 year and 2 years postoperatively. Results: BMI decreased from 47.5 � 6.2 kg/m2 to 33.2 � 5.8 kg/m2 (P <0.001). Plasma ghrelin levels were significantly increased 1 year (P <0.05) and 2 years (P <0.02) postoperatively. Fasting plasma leptin and insulin levels were significantly lower at 2 years after surgery (P <0.001). Conclusion: After gastric restrictive surgery, ghrelin levels increased, in contrast to the reported fall in ghrelin levels after gastric bypass. This difference in ghrelin levels between these operations may be the k ey to understanding the superiority of gastric bypass in sustaining weight loss compared with restrictive surgery.

Despite the current opinion that leptin can no longer be seen as a hormone which could be used therapeutically to prevent an energy surplus (it rather protects the organism for an energy deficit), leptin may still have an impact in clinical medicine. Leptin was shown to have several important functions. The pleiotropic properties of leptin include a regulatory function in the immune system. Reviewing the effects of leptin on different parts of the immune system reveals that the immune system is deregulated in an environment low in leptin. A strong reduction in leptin levels occurs in situations of starvation as seen after bariatric surgery. We postulate the hypothesis that the starvation-induced postoperative decrease of leptin is causative of the more serious course of complications observed after bariatric surgery.

Objective: To determine the variation in preadipocyte isolation procedure and to assess the number and function of preadipocytes from subcutaneous and omental adipose tissue of obese individuals. Research Methods and Procedures: The preadipocyte number per gram of adipose tissue in the abdominal‐subcutaneous and abdominal‐omental adipose stores of 27 obese subjects with a BMI of 44 ± 10 kg/m2and an age of 40 ± 9 years was determined. Results: The assessment of the preadipocyte number was found to be labor intensive and error prone. Our data indicated that the number of stromal vascular cells (SVCs), isolated from the adipose tissue by collagenase digestion, was dependent on the duration of collagenase treatment and the size and the origin of the biopsy. In addition, the fat accumulation and leptin production by differentiated SVCs were dependent on the number of adherent SVCs (aSVCs) in the culture plate and the presence of proteins derived from serum and peroxisome proliferator‐activated receptor ligands. Discussion: Using our standardized isolation and differentiation protocol, we found that the number of SVCs, aSVCs, leptin production, and fat accumulation still varied considerably among individuals. Interestingly, within individuals, the number of SVCs, aSVCs, and the leptin production by differentiating aSVCs from both the subcutaneous and the omental fat depots were associated, whereas fat accumulation was not. In obese to severely obese subjects, differences in BMI and age could not explain differences in SVCs, aSVCs, leptin production, and fat accumulation.

Gram-negative sepsis is a potentially fatal clinical syndrome characterized by a proinflammatory response (tumor necrosis factor-) to bacterial (endo)toxins and gut barrier function loss. Recently, we found that high-fat enteral nutrition protects against late bacterial translocation in a model of hemorrhagic shock in rats. However, the basis for this protection is unknown. We hypothesized that the observed protection is the result of an early inhibition of endotoxin and the subsequent inflammatory response resulting in a preserved gut barrier function. Sprague–Dawley rats were divided into a group that was starved overnight (HS-S), fed with a low-fat enteral diet (HS-LF) or fed wih a high-fat enteral diet (HS-HF), and subsequently subjected to a nonlethal hemorrhagic shock. Ninety minutes after hemorrhage, arterial endotoxin significantly decreased in HS-HF rats (4.0 ± 0.6 pg/mL) compared with HS-LF rats (10.7 ± 0.9 pg/mL, P0.002) and HS-S rats (15.2 ± 2.2 pg/mL P0.001). Interestingly, arterial tumor necrosis factor- was also decreased in HS-HF rats (17.9 ± 10.4 pg/mL) compared with HS-LF (83.5 ± 16.7 pg/mL, P< 0.01) and HS-S rats (180.9 ± 67.9 pg/mL, P< 0.02). Loss of tight junction structure (ZO-1) observed in ileum and colon of control hemorrhagic shock rats was prevented in HS-HF rats. In parallel, intestinal barrier function was preserved in HS-HF rats, evidenced by a reduced permeability to horseradish peroxidase (P< 0.05), less bacterial invasion, and a 10-fold reduction of bacterial translocation early after hemorrhagic shock. This report describes a new strategy to nutritionally prevent endotoxemia, the subsequent inflammatory response and gut barrier failure following hemorrhagic shock. High-fat enteral nutrition requires further evaluation as an intervention to prevent a potentially fatal systemic inflammatory response in patients at risk for sepsis.

We report on the design, realization, and characterization of a four-channel integrated optical Young interferometer device that enables simultaneous and independent monitoring of three binding processes. The generated interference pattern is recorded by a CCD camera and analyzed with a fast-Fourier-transform algorithm. We present a thorough theoretical analysis of such a device. The realized device is tested by monitoring glucose solutions that induce well defined phase changes between output channels. The simultaneous measurement of three different glucose concentrations shows the multipurpose feature of such devices. The observed errors, caused by the mismatching of spatial frequencies of individual interference patterns with those determined from the CCD camera, are reduced with different reduction schemes. The phase resolution for different pairs of channels was ∼1 × 10^-4 fringes, which corresponds to a refractive-index resolution of ∼8.5 × 10^-8. The measured sensitivity coefficient of the phase change versus refractive-index change of ∼1.22 × 10^3 × 2π agrees well with the calculated coefficient of ∼1.20 × 10^3 × 2π.

Three new coordination polymers have been prepared by hydrothermal reaction of squaric acid, pyrazine and the metal halides FeCl2 · 4H2O, CoBr2and NiBr2. In their crystal structures the metal atoms are coordinated by four water molecules and two pyrazine ligands within slightly distorted octahedra. The pyrazine ligands connect the metal atoms via µ-N,N’-coordination to linear chains which are connected via hydrogen bonding. The squarate dianions are not coordinated to the metal atoms and are located between the chains. The thermal behaviour of all compounds was investigated using TG-DTA-MS measurements. A complex behaviour for all compounds is found and the decomposition temperatures increase from Fe to Ni. From these investigations there are no hints for the occurrence of stoichiometric intermediate compounds.

Recently significant success has emerged from exciting research involving chromatin stretching using optical tweezers. These experiments, in which a single chromatin fibre is attached by one end to a micron-sized bead held in an optical trap and to a solid surface or second bead via the other end, allows manipulation and force detection at a single-molecule level. Through force-induced stretching of chromatin, mechanical properties, specific inter-molecular bond strengths and DNA–protein association and dissociation kinetics have been determined. These studies will be extremely fruitful in terms of understanding the function of chromatin structure and its dynamics within the cell.

We developed the CellTracks cell analysis system that, similar to flow cytometry, yields multiparameter information by which the cells can be differentiated. We describe the implementation of a laser scanning imaging method in the system. Image analysis of the cells improves the specificity of cell classification, especially in cases where the particular cells are found relatively infrequently and one has to discriminate between artifacts and real events. Fluorescent images of immunomagnetically labeled and aligned cells are obtained by passing the cells through a laser focus. The laser focus is smaller than the objects and subsequent frames captured by a regular surveillance CCD camera with a frame grabber board represent different parts of the cells. Complete images of the cells are constructed by shifting each image with respect to each other and adding individual pixel values. The power of combining a fluorescent image with multiparametric data is demonstrated by imaging fluorescent and magnetically labeled beads and cells. The image gives additional information about the dye distribution across the objects. Changes in dye distribution as a function of time were observed in leukocytes labeled with the red fluorescent label, Oxazine750, which are imaged at different time intervals. An imaging technique implemented in the CellTracks system provides high-resolution fluorescent images of events previously identified by the system. The images of the fluorescent cells enhance the ability to classify rare events. Cytometry 47:248–255, 2002. © 2002 Wiley-Liss, Inc.

A cell analysis system was developed to enumerate and differentiate magnetically aligned cells selected from whole blood. The cellular information extracted is similar to the readout of musical information from a compact disk (CD). Here we describe the optical design and data processing of the system. The performance of the system is demonstrated using fluorescent-labeled cells and beads. System performance was demonstrated with 6-μm polystyrene beads labeled with magnetic nanoparticles and allophycocyanin (APC) and immunomagnetically aligned leukocytes, fluorescently labeled with Oxazine750 and CD4-APC, CD8-Cy5.5, and CD14-APC/Cy7 in whole blood. The sensitivity of the system was demonstrated using APC-labeled beads. With this system, beads containing 333 APC molecules could easily be resolved from the background. This level of sensitivity was not achievable with a commercial flow cytometer. A maximum of 20,000 immunomagnetically labeled cells could be aligned and analyzed in between 0.6 m of Ni lines, distributed over a surface area of 18 mm² and extracted from a blood volume that depended on the height of the chamber. The utility of the system was demonstrated by performing a three-color CD4-CD8-CD14 assay. We built a cell analysis system based on immunomagnetic cell selection and alignment and analysis of fluorescent signals employing CD-technology that is as good or better than current commercial analyzers. The cell analysis can be performed in whole blood or any other type of cell suspension without extensive sample preparation. Cytometry 47:173–182, 2002. © 2002 Wiley-Liss, Inc.

Recently we introduced the CellTracks cell analysis system, in which samples are prepared based on a combination of immunomagnetic selection, separation, and alignment of cells along ferromagnetic lines. Here we describe the underlying magnetic principles and considerations made in the magnetic field design to achieve the best possible cell selection and alignment of magnetically labeled cells. Computer simulations, in combination with experimental data, were used to optimize the design of the magnets and Ni lines to obtain the optimal magnetic configuration. A homogeneous cell distribution on the upper surface of the sample chamber was obtained with a magnet where the pole faces were tilted towards each other. The spatial distribution of magnetically aligned objects in between the Ni lines was dependent on the ratio of the diameter of the aligned object and the line spacing, which was tested with magnetically and fluorescently labeled 6 μm polystyrene beads. The best result was obtained when the line spacing was equal to or smaller than the diameter of the aligned object. The magnetic gradient of the designed permanent magnet extracts magnetically labeled cells from any cell suspension to a desired plane, providing a homogeneous cell distribution. In addition, it magnetizes ferro-magnetic Ni lines in this plane whose additional local gradient adds to the gradient of the permanent magnet. The resultant gradient aligns the magnetically labeled cells first brought to this plane. This combination makes it possible, in a single step, to extract and align cells on a surface from any cell suspension. Cytometry 47:163–172, 2002. © 2002 Wiley-Liss, Inc.

Acute pancreatitis is a disease with varying severity. Patients with the mild form do not require nutritional support because oral intake is resumed rapidly. Studies on nutritional support in acute pancreatitis have included patients with both mild and severe disease. In this heterogeneous group, total parenteral nutrition did not improve outcome compared with no nutrition at all. This is caused in part by an increase in septic complications during total parenteral nutrition. Likewise, no benefit from enteral nutrition was observed compared with no nutrition, probably because the group was heterogeneous or because nutritional goals were not achieved. Patients with severe acute pancreatitis become profoundly catabolic. This group undoubtedly requires nutritional support to treat undernutrition. The limited available data indicate that enteral nutrition, if well tolerated, is superior to parenteral nutrition for patients with severe acute pancreatitis. Based on current knowledge, a combination of early total parenteral nutrition and enteral nutrition is advisable as soon as enteral nutrition is tolerated. Monitoring of gut function is crucial in this situation.

Single chromatin fibers were assembled directly in the flow cell of an optical tweezers setup. A single λ phage DNA molecule, suspended between two polystyrene beads, was exposed to a Xenopus laevis egg extract, leading to chromatin assembly with concomitant apparent shortening of the DNA molecule. Assembly was force-dependent and could not take place at forces exceeding 10 pN. The assembled single chromatin fiber was subjected to stretching by controlled movement of one of the beads with the force generated in the molecule continuously monitored with the second bead trapped in the optical trap. The force displayed discrete, sudden drops upon fiber stretching, reflecting discrete opening events in fiber structure. These opening events were quantized at increments in fiber length of ∼65 nm and are attributed to unwrapping of the DNA from around individual histone octamers. Repeated stretching and relaxing of the fiber in the absence of egg extract showed that the loss of histone octamers was irreversible. The forces measured for individual nucleosome disruptions are in the range of 20–40 pN, comparable to forces reported for RNA- and DNA-polymerases.

Atomic force microscopy (AFM) is nowadays frequently applied to determine interaction forces between biological molecules. Starting with the detection of the first discrete unbinding forces between ligands and receptors by AFM only several years ago, measurements have become more and more quantitative. At the same time, theories have been developed to describe and understand the dynamics of the unbinding process and experimental techniques have been refined to verify this theory. In addition, the detection of molecular recognition forces has been exploited to map and image the location of binding sites. In this review we discuss the important contributions that have led to the development of this field. In addition, we emphasize the potential of chemically well-defined surface modification techniques to further improve reproducible measurements by AFM. This increased reproducibility will pave the way for a better understanding of molecular interactions in cell biology.

Sepsis and organ failure remain the main cause of death on the ICU. Sepsis is characterized by a severe inflammatory response, in which plateletactivating factor PAF is considered to play an important role. This study investigated whether treatment with the PAFantagonist TCV309 reduces morbidity and mortality in patients with septic shock. The study was conducted as a doubleblind, randomized, placebo controlled multicenter study. The included patients had to fulfill the SIRS criteria with a clinical suspicion of infection, an admission APACHE II score greater than 15, and shock, defined as a mean arterial pressure <70 mmHg andor a decrease ≥40 mmHg despite adequate fluid resuscitation. Patients received 1.0 mgkg TCV309 or placebo, twice daily, intravenously during 14 days. The prospectively set goals were MOF score, recovery from shock, mortality, and assessment of the safety of the medication. A total of 98 patients were included of which 97 were analyzed on an intentiontotreat basis. The overall survival at day 56 of TCV309 treated patients was similar compared to placebo treated patients 51.0 vs. 41.7, P0.47. In contrast, the mean percentage of failed organs per patient present after 14 days in the TCV309 treated patients was significantly lower compared to the placebo treated patients 11.9 vs. 25.1, P0.04, leading to a reduced need for vasopressors, dialysis, and ventilatory support. Furthermore, the mean APACHEII score during treatment with TCV309 was significantly lower and the number of patients recovered from shock after day 14 was significantly higher in the TCV309 treated patient group 232 vs. 929, P0.01. The number of adverse events was not significantly different between the TCV309 and placebo treated patients. TCV309 did not change overall mortality of septic shock, however a substantial reduction in organ dysfunction and morbidity, frequently associated with septic shock was achieved, without significant adverse events.

We have obtained new evidence for the occurrence of intracellular NADPH-oxidase activity in neutrophilic and eosinophilic granulocytes upon stimulation with phorbol myristate acetate (PMA). PMA activation leads to a partial translocation of cytochrome b558 from the membranes of the specific granules to the plasma membrane. It was suggested that NADPH-oxidase activity only takes place in the plasma membrane, leading to an extracellular release of oxygen metabolites because cellular self-destruction can be avoided in this way. The effects of PMA activation were indirectly studied in recent experiments employing scavengers of extracellular superoxide anion and hydrogen peroxide, and support for intracellular NADPH-oxidase activity was obtained. In this paper we use Raman microspectroscopy as a direct method to study intracellular molecular reactions that result from cellular triggering by PMA. The molecular specificity of this microscopic method enables us to show that intracellular reduction of both myeloperoxidase (MPO) and cytochrome b558 occurs in neutrophilic granulocytes. Control measurements with cytochrome b558-deficient neutrophilic granulocytes did not show a reduction of intracellular MPO. This is direct support for the occurrence of intracellular NADPH-oxidase activity in organelles that must be in close contact with the azurophilic granules that contain MPO. Furthermore, a comparison was made with chemical reactions occurring in eosinophilic granulocytes after activation with PMA. Moreover, in these cells an intracellular reduction of eosinophil peroxidase was observed.