Alternative pathway activation of complement in laparoscopic and open rectal surgery.

The study was designed to investigate whether complement is activated in patients subject to rectal surgery and whether the choice of surgical technique (open or laparoscopic) has any impact on the activation of complement. Our hypothesis is that laparoscopic surgery leads to a lower-level activation of complement than open surgery. Patients (n = 24) subject to rectal surgery owing to rectal cancer were included. The study was prospective and randomized. The patients were randomized to either laparoscopic surgery (n = 12) or open surgery (n = 12). Blood samples for determination of complement activation (C4d, Bb, C3bc and the terminal C5b-9 complex TCC) were drawn before start of surgery (T0) and at the following time-points after start of surgery: 180 min (T1), 360 min (T2), 24 h (T3) and 3-5 days (T4). A significant increase in the alternative pathway activation product Bb and in the terminal pathway activation product TCC was seen over time in both groups (P < 0.001). Bb peaked early (T1) and returned to baseline levels post-operatively, whereas TCC increased steadily with maximum values in the late post-operative period. The plasma concentrations of C4d and C3bc decreased significantly in both groups at T1 and T2 and returned to baseline levels at T4. There was no significant difference between the groups. Rectal surgery causes activation of the complement system. Complement is activated through the alternative pathway. Results mostly showed no significant differences between laparoscopic and open rectal surgery apart from lower levels of factor Bb in the former group in the perioperative period.

Measuring quality in colorectal cancer surgery in low- and middle-income countries: The Clavien-Dindo classification in a Sri Lankan cohort.

Background: The colorectal cancer (CRC) incidence is increasing in low- and middle-income countries (LMICs) as part of an ongoing epidemiological transition. Surgery is the main treatment and surgical services are scaled up to meet the need. This warrants the establishment of frugal systems to measure safety and quality of surgical care that are tailored for low-resource settings. The aim of this study was to test the applicability of the Clavien-Dindo classification (CDC) for measurement of surgical complications in an LMIC setting where medical records are paper-based. Material and methods: 88 patients who underwent CRC resection at Colombo South Teaching Hospital, Sri Lanka, from January 2017 to January 2020 were included. Medical records were retrospectively reviewed for postoperative complications and the severity was graded using the CDC. Results: One or more postoperative complications (CDC ≥ grade II) occurred in 45.5% (n = 40) of the patients. The complications were distributed as grade II n = 46, grade III n = 3, grade IV n = 2 and grade V n = 0. The most common complication (22.7%, n = 20) was postoperative anemia treated with blood transfusion. The second most common complication was incisional surgical site infection (11.4%, n = 10). Conclusion: Postoperative outcome could be evaluated by using the CDC in a Sri Lankan facility based on retrospective review of medical records. This suggests that the CDC is a feasible standardized system appropriate for measuring surgical quality also in other LMICs. Identified fields for possible quality improvement at the study site were to limit blood transfusions and minimize treatment with antibiotics.

Rapid migration of thymic emigrants to the colonic mucosa in ulcerative colitis patients.

Inflammatory bowel disease (IBD) is associated with imbalances of the local intestinal immune responses, with dysregulated CD4(+) T cells contributing to the chronic inflammation. Having demonstrated altered T cell maturation in the thymus in two different mouse models of colitis, we set out to investigate whether abnormalities in T cell maturation is present in patients with ulcerative colitis (UC) or Crohn's disease (CD). Specimens were obtained from peripheral blood (CD; n = 14, UC; n = 22), colon and small intestinal specimens (CD; n = 6, UC; n = 13). As controls, peripheral blood specimens were obtained from healthy volunteers, patients with adenocarcinomas (n = 18) and colonic specimens from patients with adenocarcinomas (n = 14). Recent thymic emigrants were estimated by analysis of the normalized ratio of T cell receptor excision circles (TRECs) by real-time polymerase chain reaction (PCR). The frequency of naive- and proliferating T lymphocytes and markers of extrathymic T cell maturation in the mucosa was analyzed by flow cytometry and real time-PCR. TREC levels in peripheral blood T lymphocytes were similar between IBD patients and controls. In contrast, UC patients demonstrated significantly increased levels of TRECs both in intraepithelial and lamina propria lymphocytes from the colonic mucosa compared to patients with adenocarcinomas and CD. However, markers for extrathymic T cell maturation in the mucosa were not different between controls and IBD patients. The increased TREC levels in mucosal but not peripheral blood lymphocytes in UC patients in the absence of increased extrathymic maturation in situ in the mucosa together demonstrate that recent thymic emigrants are recruited rapidly to the inflamed mucosa of these patients.

COLOR II. A randomized clinical trial comparing laparoscopic and open surgery for rectal cancer.

INTRODUCTION: Laparoscopic resection of rectal cancer has been proven efficacious but morbidity and oncological outcome need to be investigated in a randomized clinical trial. TRIAL DESIGN: Non-inferiority randomized clinical trial. METHODS: The COLOR II trial is an ongoing international randomized clinical trial. Currently 27 hospitals from Europe, South Korea and Canada are including patients. The primary endpoint is loco-regional recurrence rate three years post-operatively. Secondary endpoints cover quality of life, overall and disease free survival, post-operative morbidity and health economy analysis. RESULTS: By July 2008, 27 hospitals from the Netherlands, Belgium, Germany, Sweden, Spain, Denmark, South Korea and Canada had included 739 patients. The intra-operative conversion rate in the laparoscopic group was 17%. Distribution of age, location of the tumor and radiotherapy were equal in both treatment groups. Most tumors are located in the mid-rectum (41%). CONCLUSION: Laparoscopic surgery in the treatment of rectal cancer is feasible. The results and safety of laparoscopic surgery in the treatment of rectal cancer remain unknown, but are subject of interim analysis within the COLOR II trial. Completion of inclusion is expected by the end of 2009. TRIAL REGISTRATION: Clinicaltrials.gov, identifier: NCT00297791 (www.clinicaltrials.gov).

Body awareness therapy: a new strategy for relief of symptoms in irritable bowel syndrome patients.

AIM: To compare irritable bowel syndrome (IBS) patients with apparently healthy persons and to evaluate body awareness therapy, which is a physiotherapeutic remedy focusing on normalising tensions in the body, for the treatment of IBS with the hypothesis that altered body tension is associated with the syndrome. METHODS: Twenty-one IBS patients received body awareness therapy two hours weekly for 24 wk. At baseline as well as after 12 and 24 wk, they underwent examinations including resource oriented body examination in combination with body awareness scale evaluation and filled in gastrointestinal and psychological symptom questionnaires. Saliva cortisol was analysed. A group of 21 apparently healthy persons underwent the same examinations once. RESULTS: Compared to the apparently healthy group, IBS patients scored higher at baseline for gastrointestinal and psychological symptoms. They showed more often alterations in normal body tension patterns, as well as deviating cortisol slopes in saliva. After 24 wk of body awareness therapy, their gastrointestinal and psychological symptoms were reduced overall. Somatic symptoms decreased in parallel with depressive symptoms. Whole body pain score decreased, coping ability as well as biochemical stress markers improved. CONCLUSION: IBS patients scored higher for gastrointestinal and psychological symptoms, and presented with altered biochemical stress markers. Their body tension deviated compared to healthy controls. Furthermore, body awareness therapy gave relief of both somatic complaints, psychological symptoms and normalised body tension. These findings indicate that distorted tension constitutes an important part of the symptoms in IBS.

Accelerated acute rejection of the intestinal graft in CD28-deficient mice.

OBJECTIVE: Multiple in vivo studies have shown that the pace and severity of graft rejection is little or not at all changed by deleting CD28 molecules in the recipient. These findings contrast with the effects of monoclonal antibody therapy aimed the same costimulatory target. The objective of the present study was to evaluate how the acute rejection process is affected in CD28-deficient mice using a fully allogeneic, highly immunologically reactive transplant model. METHODS: Heterotopic vascularized small bowel transplants were performed in 24 recipient mice divided into 4 groups: 2 wild-type and 2 knockout groups. Each group consisted of 5 to 7 animals in which BalbC mice were used as intestinal donors to either wild-type C57BL6 or C57BL6 background CD28-deficient recipient mice. Selected endpoints were 3 and 6 postoperative days (POD). Intestinal rejection was evaluated by mucosal laser Doppler flowmetry (expressed in perfusion units) and histology (expressed in rejection grades). RESULTS: Acute rejection occurred in both wild-type and CD28-deficient groups. At POD 3, no significant difference was noted between groups in terms of mucosal perfusion and histology. At POD 6, significant differences in graft mucosal perfusion and histology revealed a more aggressive rejection in the CD28-deficient group compared to the wild-type group. CONCLUSIONS: The present study showed that the severity of intestinal graft rejection responses was amplified by deleting CD28 molecules. Together with data from other studies, these results suggest a different pattern of distribution and/or activation of CD28/B7 receptors in various organs.

Heterotopic uterine transplantation by vascular anastomosis in the mouse.

A method of heterotopic uterine transplantation was developed in the mouse as a model system for studies of uterine function and transplant immunology of the uterus. The model involved transplantation of the right uterine horn and the cervix by vascular anastomosis to a donor animal with the intact native uterus remaining in situ. F1-hybrids of inbred C57BL/6 x CBA/ca (B6 CBAF1) mice of 6-8 weeks of age (n=42) were used. The specific pelvic vascular anatomy of these mice was first examined by intra-aortal injection of a two-component silicon-rubber curing agent. The surgery of the donor animal involved microsurgical isolation of the right uterine horn and the cervix, with preserved vascular supply from the aorta through the right uterine artery. After isolation of the uterine horn with vascular supply and venous drainage, including approximately 3 mm of the inferior vena cava and aorta, the organ was put on ice. The recipient animal was prepared by exposing and mobilizing the infrarenal part of the aorta and the vena cava. The grafted uterus was placed in the abdomen on the left side and the aorta and vena cava of the graft were anastomosed end-to-side to the aorta and vena cava of the recipient animal with 11-0 sutures. The total time for these procedures declined with time and was 125+/-4 min for the last 28 operations. Viability of the uterus was confirmed, several days later, by demonstrating a blood flow similar to that of the native uterus, and histology of the grafted uterus demonstrated normal morphology, including intact ultrastructure of the endothelial cells. The overall survival rate of the recipient animals increased with learning from approximately 40% in animals 1-21 to 71% in animals 22-42. The proportion of viable grafts, as judged by normal blood flow and histology among the surviving mice was 25% in animals 1-21 and 87% in animals 22-42. An undisturbed function of the transplanted uterus horn was finally demonstrated by its ability to implant inserted blastocysts and to carry pregnancy with fetal weight being similar to that of fetuses in the native uterus and controls. In conclusion, this is the first report of successful transplantation of the uterus with proven functionality in the mouse. The model should be useful for many aspects of research in uterine physiology and pathophysiology.

Factors reducing mortality in typhoid ileal perforation.

A retrospective study of 42 patients treated surgically for typhoid ileal perforation found the mortality to be 24%. The study suggested that general peritonitis was the main cause of death in typhoid ileal perforation. However, the high mortality was significantly improved by adequate preoperative rehydration. The combination of chloramphenicol/gentamicin/metronidazole was superior to other antibiotic regimes. No mortality was recorded with this triple drug therapy.

National registry of patients with short bowel syndrome.

In Sweden, with a population of 9 million, there are an estimated 400 people suffering from short bowel syndrome (SBS). These patients are dispersed throughout the country and are cared for mostly by ordinary health-care providers, who have limited experience with SBS. During the last decade the patients and their organizations have expressed increasing concerns about competent and equally distributed care. Against this background, the National Association of People with Stomach and Bowel Diseases, together with a faculty of experts in the field, created a web-based registry to facilitate follow-up and to improve quality of care of SBS patients. Their homepage, www.swegir.com/KTS, is based on the Swedish General Internet Registry administered by the Swedish Society of Surgery and Gastroenterology. Its homepage includes some information on SBS for the public, as well as links to a health manual on SBS and different associations. Behind a fire-wall, health professionals have access to an on-line registry for follow-up of patients with short bowel syndrome. In the future, on-line statistics regarding the number of complications, for example, will be available from the registry. Patients who would benefit from small bowel transplantation could also be identified.

Intragraft heat shock protein-60 expression after small bowel transplantation in the mouse.

The time course of heat shock protein 60 (hsp 60) expression after intestinal transplantation in syngeneic and allogeneic combination was correlated with the degree of rejection. Hsp 60 expression was assessed by immunostaining; rejection degree was established by histologic examination on posttransplantation days 1, 3, 6, and 8. No signs of rejection occurred in syngeneic grafts at any time. In the allogeneic setting, rejection was absent in all but 1 case on postoperative day 3. Three days later moderate rejection was evident based on focal crypt destruction and focal mucosal ulceration, whereas at postoperative day 8 extensive mucosal sloughing was the dominant feature, consistent with advanced rejection. Hsp 60 remained undetectable in the syngeneic setting at all times. In allografts, hsp 60 was initially expressed on posttransplant day 3, increasing synchronously with the progression of rejection at days 6 and 8. Hsp 60 expression was localized almost exclusively to the crypt area and the lower third of the villi. In conclusion, the rejection of murine allogeneic intestinal grafts is characterized by a progressive expression of hsp 60 in the epithelium.

High prevalence of vitamin D deficiency and osteoporosis in out-patients with intestinal failure.

BACKGROUND: In intestinal failure, specific nutrient deficiencies especially for fat-soluble vitamins can be expected in addition to energy-protein malnutrition. We report serum levels of fat soluble vitamins, and bone density in out-patients with intestinal failure (IF). METHODS: 106 outpatients with IF were assessed during routine visits. 78 patients underwent DXA-scan for bone density. Vitamin D levels < 50 nmol/l were defined as deficiency, and 75-150 as optimal. Vitamin A and E deficiencies were defined as <1.0 and <14 µmol/l respectively. INR ≥ 1.2 without liver disease or anti-vitamin K therapy was classified as vitamin K deficiency. RESULTS: Mean serum vitamin D level was 45 nmol/l at first visit, and 64 nmol/l at follow up (n = 76, p = 0.0001 by paired t-test). Overall prevalence of vitamin D deficiency was 67%. Only 12% of all patients had optimal D-vitamin status. 88% of assessed patients had low bone density. 12% had subnormal vitamin A levels and 25% had subnormal vitamin E levels. 32% had abnormal INR values. At follow up 34% remained vitamin D deficient whereas 29% had optimal levels. By oral substitution, vitamin A and E status were normalised, and K status improved. CONCLUSION: Vitamin D deficiency and osteoporosis are common in outpatients with intestinal failure, and should be adequately substituted.

Rejection patterns in allogeneic uterus transplantation in the mouse.

BACKGROUND: Transplantation of the uterus in the mouse has been developed as a model system for research towards human uterine transplantation. Previous studies in a mouse model have demonstrated that a syngeneic uterus transplant can give rise to normal offspring. The aim of this study was to characterize the time course of rejection in a fully allogeneic mouse uterus transplantation model. METHODS: Uteri of BALB/c mice were transplanted to a heterotopic position in C57BL/6 recipients, whose native uteri were left in situ. The blood flow of the uteri, their gross appearance and general histology and the density of T-lymphocytes were examined on postoperative days 2-28. RESULTS: Macroscopic signs of rejection were apparent from day 5. At the light microscopy level, minimal inflammatory changes were seen from day 5 and massive inflammation was seen from day 10 to day 15. At day 28, necrosis and fibrosis were seen. The density of T-lymphocytes (CD3(+)) was increased in the grafted uterus from day 2 in the myometrium and from day 5 in the endometrium. Blood flow in the grafted uteri was reduced from day 15. CONCLUSION: A murine model to study rejection of allogeneic uterus transplants was characterized. Signs of rejection were seen from day 2 to day 5 and severe rejection was seen from day 10 to day 15. The data will be useful in future studies on immunosuppressants in this model.

Short-term ischaemic storage of human uterine myometrium--basic studies towards uterine transplantation.

BACKGROUND: Transplantation of the uterus has been suggested as a possible future treatment of absolute uterine infertility. The tolerability of human uterine tissue to cold ischaemic storage was tested in the present study. METHODS: Small tissue samples of human uteri were subjected to cold (4 degrees C) ischaemia (6 and 24 h) in Ringer acetate (RIN), the intracellular-like University of Wisconsin solution (UW) or the extracellular-like Perfadex solution (PER). The ability of myometrial strips to contract, histology by light and electron microscopy as well as tissue concentrations of glutathione, ATP and protein were used as parameters to detect cold ischaemic injuries. RESULTS: Contractile ability and response to prostaglandin F(2alpha) (PGF(2alpha)) was better preserved after 6 h cold ischaemia in UW and PER in comparison with the other groups. Histological examination did not reveal any major changes after 6 and 24 h cold ischaemic storage in UW and PER solutions, while specimens stored in RIN for 24 h displayed degenerative changes on the electron microscopy level. UW and PER preserved ATP concentrations significantly better than RIN. Myometrium stored in UW contained more total glutathione but also a larger proportion of oxidized glutathione than specimens stored in RIN and PER. Protein concentrations did not change with storage time in any of the solutions. CONCLUSIONS: The results show that human uterine myometrial tissue is resistant towards cold ischaemia for at least 6 h if stored in UW and PER solutions.

[Research in developing countries: will leprosy vaccine be produced in Nepal?]. / Forskning i u-land: kommer lepravaccinet att framställas i Nepal?

Clinical, bacteriological and immunological research into leprosy is pursued at the Anandaban Leprosy Hospital on the outskirts of Kathmandu in Nepal with support from the World Health Organization and research laboratories in inter alia Holland and Australia. The bacteriological part consists of research into cultivation procedures on mice for diagnosis of Mycobacterium Leprae resistance to dapsone and rifampicin. Serological studies have also yielded M. Leprae-specific antibodies, a method which, it is hoped, will be further developed as an aid to diagnosis and follow-up of treatment. Research concerning a leprosy vaccine is also in progress.

Integrated intestinal capacity and nutritional status following small bowel transplantation.

Successful small bowel transplantation requires normal functional capacity of the graft and unaltered metabolism of the host. Weight gain and wet weight of muscle groups and intra-abdominal fat pads were compared between transplanted, sham-operated, short bowel-operated, and normal rats that were fed either standard chow or fat-enriched (15%) pellets. Weight gain and wet weight of muscle groups and fat pads for the control, transplanted, and sham-operated rats were identical, while short bowel animals showed reduced weight. Transplanted rats receiving fat-enriched food had lower wet weight of fat pads than control animals on the high-fat diet. We conclude that small bowel transplantation makes it possible to overcome the intestinal failure associated with short bowel syndrome, leading to overall normal weight gain and development of the recipient. However, altered fat metabolism, reflected in changed body composition, was observed in transplanted animals on the high-fat diet.

Effects of hypoperfusion on small-intestinal mucosal morphology and intestinal barrier function.

BACKGROUND: Our aim was to evaluate in vitro perfusion of the rat small intestine as a method to study the pathophysiology of hypoperfusion and to minimize its adverse effects. METHODS: Rat small intestine was perfused for 24 h using RPMI + albumin for the lumen and RPMI + erythrocyte concentration (Hct 20%) in the vasculature, at 22 degrees C, 15 degrees C, 10 degrees C, and 6 degrees C. The outcome was evaluated by histologic grading and permeability for polyethylene glycol 900 (PEG 900). RESULTS: Perfusion of the intestine at 6 degrees C resulted in lower degree of mucosal damage and permeation of PEG 900, compared with storage and perfusion at higher temperatures. CONCLUSIONS: In vitro perfusion of the small intestine, using histologic grading of mucosal lesions and permeation of PEG 900 as indicators, is one method that can be applied to study the pathophysiology of intestinal hypoperfusion. Perfusion at 6 degrees C significantly reduced the level of damage to the intestinal mucosa.

Migration of host and donor T cells in small bowel transplantation.

In this study migration of host and donor CD4+ and CD8+ T cells in a fully allogeneic model was described and compared with the migration pattern in a graft-versus-host reaction (GVHR) model, where the T-cell traffic in the graft served as a physiological control. Heterotopic small bowel transplantations were performed in a rat model, with animals being sacrificed on postoperative days (POD) 2, 3, 4, 5, and 7. Graft and host mesenteric lymph nodes were harvested, homogenized, and stained with monoclonal antibodies against MHC class I, CD4+, and CD8+ antigens. The host and donor T cell migration patterns were studied using a double-staining flow cytometric technique. We found that during the development of rejection, the normal physiological circulation of graft and host T cells was disrupted. In the graft of the allogeneic model, a shift from host cell to graft cell dominance occurred on POD 3-4. This change in migration pattern coincided in the host with a 6% peak in graft cell infiltration, which disappeared on POD 7. These patterns of T-cell migration may be further explored for diagnostic purposes.

Pregnancy in transplanted mouse uterus after long-term cold ischaemic preservation.

BACKGROUND: The aim of this study was to evaluate the viability of the transplanted murine uterus after cold ischaemic preservation. METHODS: Uteri of mice (6-8 weeks old) were isolated and kept at 4 degrees C in vitro for 24 or 48 h in 0.154 mol/l NaCl or University of Wisconsin (UW) solution. Viability was evaluated by assessment of morphology and contractility in vitro. Furthermore, uteri were transplanted by vascular anastomoses to syngeneic recipients after 24 or 48 h cold ischaemic preservation in UW solution and morphology, blood flow and capacity to implant transferred blastocysts were assessed 2 weeks later. RESULTS: Uteri that had been preserved for 24 h exhibited normal morphology but after 48 h preservation minimal degenerative changes were seen. Spontaneous contractions occurred in uteri after 24 h as well as 48 h cold ischaemic preservation and prostaglandin F(2alpha)-stimulated responses were preserved. Blood flow and morphology were normal 2 weeks after transplantation in uteri preserved for 24 h, while grafts preserved for 48 h had a decreased blood flow and morphology showed total necrosis of the transplants. Transplanted uteri that had been preserved for 24 h developed pregnancies (in five out of six animals) after embryo transfer, with offspring showing normal weight and growth trajectory. CONCLUSIONS: This study shows for the first time that the mouse uterus tolerates cold ischaemic preservation and that pregnancies can be carried in transplanted uteri that have been preserved for 24 h.

Blockade of the B7-CD28 pathway by CTLA4-Ig counteracts rejection and prolongs survival in small bowel transplantation.

Allograft rejection involves T-cell activation, requiring T-cell receptor interactions with major histocompatibility complex (MHC) molecules and costimulatory signals delivered through the B7-CD28 pathway. We evaluated the effect of blocking this pathway on graft rejection and survival, in a rat experimental model of small bowel transplantation. Heterotopic small bowel transplantation was performed between PVG donor rats and DA recipient rats. The recipient animals were treated with CTLA4-Ig or irrelevant immunoglobulin (Ig)G as control and followed for 18, 30 or 90 days. The survival rate and degree of inflammation and accumulation of CD4+ T cells and macrophages were determined in the transplanted bowels. We found that administration of CTLA4-Ig significantly improved the survival rate compared to control rats: after 30 days 73% of the treated rats had survived and at 90 days 5/8 rats were still living, whereas in the control group only 2/8 rats had survived. The grafts showed preserved mucosal structure with only a mild degree of subacute inflammation and the accumulation of CD4+ T cells and macrophages was noticeably reduced in treated animals as compared to control rats. Necrosis was extensive in control rats, whereas CTLA4-Ig treated animals had grafts with at least some preserved villus morphology and no necrotic tissue. Although small bowel transplantation has proven exceptionally difficult, in this study we have shown that CTLA4-Ig treatment may provide a promising strategy to prevent rejection and induce long term tolerance and graft survival.