Faecal microbiota signatures of IBD and their relation to diagnosis, disease phenotype, inflammation, treatment escalation and anti-TNF response in a European Multicentre Study (IBD-Character).

METHOD: We examined faecal samples, using the GA-map™ Dysbiosis Test, to associate gut microbiota composition with Crohn's disease (CD) and ulcerative colitis (UC) and to identify markers for future biomarker identification. We conducted a prospective case-control study (EU-ref. no. 305676) in an inception cohort of 324 individuals (64 CD, 84 UC, 116 symptomatic non-IBD controls and 44 healthy controls) across five European centres and examined 54 predetermined bacterial markers. We categorized patients according to the Montreal Classification and calculated the dysbiosis index (DI). Non-parametric tests were used to compare groups and the Bonferroni correction to adjust for multiple comparisons. RESULTS: The fluorescent signals (FSSs) for Firmicutes and Eubacterium hallii were lower in inflammatory bowel disease (IBD) vs. symptomatic controls (p<.05). FSS for Firmicutes, Lachnospiraceae, Eubacterium hallii and Ruminococcus albus/bromii were lower, whereas the signal for Bacteroides Fragilis was higher in UC vs. symptomatic controls (p<.05). FSS was higher for Bifidobacterium spp., Eubacterium hallii, Actinobacteria and Firmicutes among patients with ulcerative proctitis, compared to extensive colitis (p<.05). In CD, we observed no association with disease location. The DI correlated with faecal-calprotectin in both CD and in UC (p<.001). In terms of treatment escalation and anti-TNF response, differences were observed for some bacterial markers, but none of these associations were statistically significant. CONCLUSION: Our data reveal that the GA-map™ Dysbiosis Test holds the potential to characterize the faecal microbiota composition and to assess the degree of dysbiosis in new-onset IBD. On the other hand, our results cannot demonstrate any proven diagnostic or predictive value of this method to support clinical decision making.

CLC and IFNAR1 are differentially expressed and a global immunity score is distinct between early- and late-onset colorectal cancer.

Colorectal cancer (CRC) incidence increases with age, and early onset of the disease is an indication of genetic predisposition, estimated to cause up to 30% of all cases. To identify genes associated with early-onset CRC, we investigated gene expression levels within a series of young patients with CRCs who are not known to carry any hereditary syndromes (n=24; mean 43 years at diagnosis), and compared this with a series of CRCs from patients diagnosed at an older age (n=17; mean 79 years). Two individual genes were found to be differentially expressed between the two groups, with statistical significance; CLC was higher and IFNAR1 was less expressed in early-onset CRCs. Furthermore, genes located at chromosome band 19q13 were found to be enriched significantly among the genes with higher expression in the early-onset samples, including CLC. An elevated immune content within the early-onset group was observed from the differentially expressed genes. By application of outlier statistics, H3F3A was identified as a top candidate gene for a subset of the early-onset CRCs. In conclusion, CLC and IFNAR1 were identified to be overall differentially expressed between early- and late-onset CRC, and are important in the development of early-onset CRC.

Serum proteomic profiling at diagnosis predicts clinical course, and need for intensification of treatment in inflammatory bowel disease.

BACKGROUND: Success in personalized medicine in complex disease is critically dependent on biomarker discovery. We profiled serum proteins using a novel proximity extension assay [PEA] to identify diagnostic and prognostic biomarkers in inflammatory bowel disease [IBD]. METHODS: We conducted a prospective case-control study in an inception cohort of 552 patients [328 IBD, 224 non-IBD], profiling proteins recruited across six centres. Treatment escalation was characterized by the need for biological agents or surgery after initial disease remission. Nested leave-one-out cross-validation was used to examine the performance of diagnostic and prognostic proteins. RESULTS: A total of 66 serum proteins differentiated IBD from symptomatic non-IBD controls, including matrix metallopeptidase-12 [MMP-12; Holm-adjusted p = 4.1 × 10-23] and oncostatin-M [OSM; p = 3.7 × 10-16]. Nine of these proteins are associated with cis-germline variation [59 independent single nucleotide polymorphisms]. Fifteen proteins, all members of tumour necrosis factor-independent pathways including interleukin-1 (IL-1) and OSM, predicted escalation, over a median follow-up of 518 [interquartile range 224-756] days. Nested cross-validation of the entire data set allowed characterization of five-protein models [96% comprising five core proteins ITGAV, EpCAM, IL18, SLAMF7 and IL8], which define a high-risk subgroup in IBD [hazard ratio 3.90, confidence interval: 2.43-6.26], or allowed distinct two- and three-protein models for ulcerative colitis and Crohn's disease respectively. CONCLUSION: We have characterized a simple oligo-protein panel that has the potential to identify IBD from symptomatic controls and to predict future disease course. Further prospective work is required to validate our findings.

Whole Blood Profiling of T-cell-Derived microRNA Allows the Development of Prognostic models in Inflammatory Bowel Disease.

BACKGROUND: MicroRNAs [miRNAs] are cell-specific small non-coding RNAs that can regulate gene expression and have been implicated in inflammatory bowel disease [IBD] pathogenesis. Here we define the cell-specific miRNA profiles and investigate its biomarker potential in IBD. METHODS: In a two-stage prospective multi-centre case control study, next generation sequencing was performed on a discovery cohort of immunomagnetically separated leukocytes from 32 patients (nine Crohn's disease [CD], 14 ulcerative colitis [UC], eight healthy controls) and differentially expressed signals were validated in whole blood in 294 patients [97 UC, 98 CD, 98 non-IBD, 1 IBDU] using quantitative PCR. Correlations were analysed with phenotype, including need for early treatment escalation as a marker of progressive disease using Cox proportional hazards. RESULTS: In stage 1, each leukocyte subset [CD4+ and CD8+ T-cells and CD14+ monocytes] was analysed in IBD and controls. Three specific miRNAs differentiated IBD from controls in CD4+ T-cells, including miR-1307-3p [p = 0.01], miR-3615 [p = 0.02] and miR-4792 [p = 0.01]. In the extension cohort, in stage 2, miR-1307-3p was able to predict disease progression in IBD (hazard ratio [HR] 1.98, interquartile range [IQR]: 1.20-3.27; logrank p = 1.80 × 10-3), in particular CD [HR 2.81; IQR: 1.11-3.53, p = 6.50 × 10-4]. Using blood-based multimarker miRNA models, the estimated chance of escalation in CD was 83% if two or more criteria were met and 90% for UC if three or more criteria are met. INTERPRETATION: We have identified and validated unique CD4+ T-cell miRNAs that are differentially regulated in IBD. These miRNAs may be able to predict treatment escalation and have the potential for clinical translation; further prospective evaluation is now indicated.

C-reactive protein: a predictive factor and marker of inflammation in inflammatory bowel disease. Results from a prospective population-based study.

BACKGROUND AND AIMS: C-reactive protein (CRP) levels are often used in the follow-up of patients with inflammatory bowel disease (IBD). The aims of this study were to establish the relationship of CRP levels to disease extent in patients with ulcerative colitis and to phenotype in patients with Crohn's disease, and to investigate the predictive value of CRP levels for disease outcome. METHODS: CRP was measured at diagnosis and after 1 and 5 years in patients diagnosed with IBD in south-eastern Norway. After 5 years, 454 patients with ulcerative colitis and 200 with Crohn's disease were alive and provided sufficient data for analysis. RESULTS: Patients with Crohn's disease had a stronger CRP response than did those with ulcerative colitis. In patients with ulcerative colitis, CRP levels at diagnosis increased with increasing extent of disease. No differences in CRP levels at diagnosis were found between subgroups of patients with Crohn's disease as defined according to the Vienna classification. In patients with ulcerative colitis with extensive colitis, CRP levels above 23 mg/l at diagnosis predicted an increased risk of surgery (odds ratio (OR) 4.8, 95% confidence interval (CI) 1.5 to 15.1, p = 0.02). In patients with ulcerative colitis, CRP levels above 10 mg/l after 1 year predicted an increased risk of surgery during the subsequent 4 years (OR 3.0, 95% CI 1.1 to 7.8, p = 0.02). A significant association between CRP levels at diagnosis and risk of surgery was found in patients with Crohn's disease and terminal ileitis (L1), and the risk increased when CRP levels were above 53 mg/l in this subgroup (OR 6.0, 95% CI 1.1 to 31.9, p = 0.03). CONCLUSIONS: CRP levels at diagnosis were related to the extent of disease in patients with ulcerative colitis. Phenotype had no influence on CRP levels in patients with Crohn's disease. CRP is a predictor of surgery in subgroups of patients with either ulcerative colitis or Crohn's disease.

The FCRL3 -169T>C polymorphism is associated with rheumatoid arthritis and shows suggestive evidence of involvement with juvenile idiopathic arthritis in a Scandinavian panel of autoimmune diseases.

BACKGROUND AND OBJECTIVES: The Fc receptor-like 3 (FCRL3) gene -169T>C single nucleotide polymorphism (SNP) has been reported to be associated with several autoimmune diseases (AIDs) in Japanese populations. However, association results in other populations have been conflicting. Therefore, we investigated this SNP in a Scandinavian panel of AIDs. METHODS: We genotyped patients with rheumatoid arthritis (RA; n = 708), juvenile idiopathic arthritis (JIA; n = 524), systemic lupus erythaematosus (SLE; n = 166), ulcerative colitis (UC; n = 335), primary sclerosing cholangitis (PSC; n = 365), Crohn disease (CD; n = 149), a healthy control group (n = 1030) and 425 trio families with type 1 diabetes (T1D). Statistical analysis consisted of case-control and family-based association tests. RESULTS: RA was associated with the C allele (odds ratio (OR) = 1.16, 95% CI 1.01 to 1.33) and the CC genotype (OR = 1.30, 95% CI 1.01 to 1.67) of the FCRL3 -169T>C SNP in our material. Suggestive evidence for association was also found for JIA (CC genotype: OR = 1.30, 95% CI 0.99 to 1.70), and clinical subgroup analysis indicated that this was connected to the polyarticular subgroup. No significant association was found with SLE, UC, CD, PSC or T1D. In patients with RA, we found no significant interaction between the FCRL3 -169T>C and PTPN22 1858C>T SNPs, nor between the FCRL3 -169CC genotype and IgM-rheumatoid factor or anti-cyclic citrullinated peptide titre levels. CONCLUSION: We found an association between the FCRL3 -169T>C SNP and RA, and suggestive evidence for involvement with JIA, in a Norwegian population. These findings lend support for a role for this SNP in RA across ethnically diverse populations, and warrant follow-up studies in JIA.

Characterization of Intestinal Microbiota in Ulcerative Colitis Patients with and without Primary Sclerosing Cholangitis.

BACKGROUND AND AIMS: There is an unexplained association between ulcerative colitis [UC] and primary sclerosing cholangitis [PSC], with the intestinal microbiota implicated as an important factor. The study aim was to compare the structure of the intestinal microbiota of patients with UC with and without PSC. METHODS: UC patients with PSC [PSC-UC] and without PSC [UC] were identified from biobanks at Oslo University Hospital, Foothills Hospital Calgary and Mount Sinai Hospital Toronto. Microbial DNA was extracted from colonic tissue and sequencing performed of the V4 region of the 16S rRNA gene on Illumina MiSeq. Sequences were assigned to operational taxonomic units [OTUs] using Quantitative Insights Into Microbial Ecology [QIIME]. Microbial alpha diversity, beta diversity, and relative abundance were compared between PSC-UC and UC phenotypes. RESULTS: In all, 31 PSC-UC patients and 56 UC patients were included. Principal coordinate analysis [PCoA] demonstrated that city of sample collection was the strongest determinant of taxonomic profile. In the Oslo cohort, Chao 1 index was modestly decreased in PSC-UC compared with UC [p = 0.04] but did not differ significantly in the Calgary cohort. No clustering by PSC phenotype was observed using beta diversity measures. For multiple microbial genera there were nominally significant differences between UC and PSC-UC, but results were not robust to false-discovery rate correction. CONCLUSIONS: No strong PSC-specific microbial associations in UC patients consistent across different cohorts were identified. Recruitment centre had a strong effect on microbial composition. Future studies should include larger cohorts to increase power and the ability to control for confounding factors.

Deviations in human gut microbiota: a novel diagnostic test for determining dysbiosis in patients with IBS or IBD.

BACKGROUND: Dysbiosis is associated with many diseases, including irritable bowel syndrome (IBS), inflammatory bowel diseases (IBD), obesity and diabetes. Potential clinical impact of imbalance in the intestinal microbiota suggests need for new standardised diagnostic methods to facilitate microbiome profiling. AIM: To develop and validate a novel diagnostic test using faecal samples to profile the intestinal microbiota and identify and characterise dysbiosis. METHODS: Fifty-four DNA probes targeting ≥300 bacteria on different taxonomic levels were selected based on ability to distinguish between healthy controls and IBS patients in faecal samples. Overall, 165 healthy controls (normobiotic reference collection) were used to develop a dysbiosis model with a bacterial profile and Dysbiosis Index score output. The model algorithmically assesses faecal bacterial abundance and profile, and potential clinically relevant deviation in the microbiome from normobiosis. This model was tested in different samples from healthy volunteers and IBS and IBD patients (n = 330) to determine the ability to detect dysbiosis. RESULTS: Validation confirms dysbiosis was detected in 73% of IBS patients, 70% of treatment-naïve IBD patients and 80% of IBD patients in remission, vs. 16% of healthy individuals. Comparison of deep sequencing and the GA-map Dysbiosis Test, (Genetic Analysis AS, Oslo, Norway) illustrated good agreement in bacterial capture; the latter showing higher resolution by targeting pre-determined highly relevant bacteria. CONCLUSIONS: The GA-map Dysbiosis Test identifies and characterises dysbiosis in IBS and IBD patients, and provides insight into a patient's intestinal microbiota. Evaluating microbiota as a diagnostic strategy may allow monitoring of prescribed treatment regimens and improvement in new therapeutic approaches.

Secondary amyloidosis in inflammatory bowel disease: a study of 18 patients admitted to Rikshospitalet University Hospital, Oslo, from 1962 to 1998.

BACKGROUND: Amyloidosis (A) is a well-known but rare complication to inflammatory bowel disease (IBD). We describe 18 patients with IBD and A, with special emphasis on clinicopathologic features and site relationships, comparing our results with previously reported cases in the world literature. METHODS: Patient records were collected from the files of the medical department at Rikshospitalet. Clinical data were compiled from records. RESULTS: Fifteen of the 18 patients had Crohn's disease (CD), 1 had ulcerative colitis (UC), one had UC preceding CD, and 1 had indeterminate colitis. There was a male preponderance of 13:5 = 2.6. Five of the patients had A at the time of diagnosis of IBD. Median time from diagnosis of IBD to A was 4 years, and A was diagnosed within 5 years after onset of IBD in 11 patients. Thirteen of the patients had suppurative complications; 12 had extraintestinal manifestations. Sixteen of the patients had been treated by bowel resection, 14 due to refractory IBD. Ten patients had been treated by renal transplantation. After 15 years of follow-up, the survival rate was 60%. CONCLUSIONS: Our findings strengthen the previous impression of an approximately 3-fold increased preponderance in males, with at least 10-fold increased frequency in CD compared with UC, and with a possible relationship to suppurative complications and extraintestinal manifestations, as well as an increased risk of having a bowel resection. The increased survival seems to be due to the introduction of renal transplantation.

Lifestyle-related factors and colorectal polyps: preliminary results from a Norwegian follow-up and intervention study.

Lifestyle-related variables are suggested to play a major role in the development of colorectal cancer (CRC). Within a 3-year follow-up and intervention study with calcium and antioxidants against growth and recurrence of colorectal polyps, supplementary studies were performed in which different aspects of lifestyle were examined. Instead of polypectomy at diagnosis, polyps <9 mm were left in situ in 116 polyp patients (50-76 years, 50% men). After 3 years, all polyps were removed and subjected to histology. Two different sets of control groups were included (all controls were age- and sex-matched and proven to be free of polyps). We applied two different methods in order to assess most exposure variables. Generally, in case-control studies, the validity of the study outcomes is high if they are similar regardless of choice of controls and methods, since bias due to these choices may affect the risk estimates. In contrast, the validity of the study outcomes is low if dependent upon these choices. Our preliminary data support the theory that different factors may be of importance in different stages of the neoplastive formation, and that lifestyle-related factors are likely to play a major role in CRC development.

Dietary factors and colorectal polyps: a case-control study.

In the case-control study we compared dietary habits among 108 patients with small (< or = 5 mm, n = 26), medium (5-9 mm, n = 48) or large (> or = 10 mm, n = 34) colorectal polyps with 35 healthy age- and gender-matched controls. A food record by weighing during 5 consecutive days was performed. The intake of fat was significantly higher among the patients, in contrast to a significantly lower intake of carbohydrate, dietary fibre and iron, compared with controls. The intake of vitamin C and calcium was shown to be lower among the patients, but this was significant only for women. There was a tendency among the patients to consume a lower-antioxidant, fibre and cereal fibre diet, and a calcium-rich and more cholesterol-rich diet with increasing size of polyps. The patients with the smallest polyps tended to consume less starch. Our results are too preliminary to draw conclusions with regard to the influence of nutritional factors on the size and growth of polyps. However, our risk factors for the presence of polyps are in agreement with previous studies. Further studies taking into account the size of the polyp are needed to corroborate our findings.

Smoking and colorectal adenomas: a case-control study.

A positive association between tobacco and colorectal adenomas has been suggested. Smoking is, however, also associated with 'poor' dietary habits, which in turn may be related to risk of adenomas. It is therefore of interest to study the relationship between smoking, diet and risk of colorectal adenomas in follow-up studies. We compared 87 adenoma cases to 35 'hospital' and 35 healthy controls (all controls were age- and sex-matched and proven to be free of adenomas). Smoking data were collected by an interview and a self-administrated questionnaire with a time interval of at least one month. After 3 years of follow-up, all polyps were removed. Our data indicate that smoking is associated with adenoma prevalence, but not necessarily with size, multiplicity, growth or recurrence of adenomas. Compared to both sets of controls, cases reported to have smoked more than 15 pack-years, or who are current smokers, had a fourfold increased frequency of adenomas (odds ratios 3.6-5.9). Smokers with adenomas had dietary habits that may also be associated with adenomas. The smoking estimates remained largely unchanged even after adjustments for dietary variables in multivariate analysis. This study lends support to the theory of an initiating role of tobacco smoke in neoplasia formation.

The relationship between faecal bile acid profile with or without supplementation with calcium and antioxidants on recurrence and growth of colorectal polyps.

Faecal bile acids (FBA) have been implicated in colon carcinogenesis. The results of case-control studies of colorectal cancer and polyp patients are, however, conflicting. The aim of this study was to examine the influence of faecal bile acids on occurrence, growth and recurrence of colorectal polyps, and to see if a mixture of calcium and antioxidants might possibly act on cancer precursors through the effect on FBA. A total of 116 polyp-bearing patients were recruited from the outpatients department. Polyps < 10 mm in diameter were left in situ and measured by annual colonoscopy for 3 years. The patients received placebo or a mixture of antioxidants and calcium carbonate, 1.6 g calcium ion daily. Faecal samples were collected annually; the first, 1 month after start of intervention, freeze dried and subjected to bile acid profile analysis. Two age and sex matched control groups were recruited (n = 35), one from healthy volunteers (healthy controls) and one from the outpatients referred for colonoscopy, with no polyps (hospital controls). Twelve of 47 patients from the healthy volunteers had polyps (healthy polyp patients). One or more adenomas were found in 93 patients. The faeces of the hospital controls had significantly higher concentrations of total and secondary bile acids than did the healthy controls. There was no difference in FBA profile between the polyp group and the hospital controls, but significantly higher concentration of total and secondary faecal bile acids in the healthy polyp patients compared with the healthy control group (P < 0.05). No increased concentration of FBA were found in the polyp patients with multiple polyps (n = 21) or previous treatment for colorectal cancer (n = 7). No associations between FBA profile and growth or recurrence of colorectal polyps were found. The polyp patients receiving active medication had higher faecal concentrations of total and secondary bile acids in the beginning of the study than at the end, in spite of a good compliance. The present study does not support bile acids as being important markers of initiation or growth of small and medium sized colorectal adenomas. In the present study the calcium and antioxidants did not seem to affect the growth or recurrence of colorectal adenomas by increased TBA excretion in the faeces.

Do undesirable effects of screening affect all-cause mortality in flexible sigmoidoscopy programmes? Experience from the Telemark Polyp Study 1983-1996.

There is substantial evidence for the beneficial effect of screening programmes aimed at reducing mortality from colorectal cancer (CRC). The effect on all-cause mortality, however, may not necessarily be beneficial. In the present study we used the follow-up results 13 years after a flexible sigmoidoscopy screening to evaluate the long-term effects of informing participants about findings at flexible sigmoidoscopy (FS) screening. There were no severe complications and there was no long-term difference in deaths related to whether there had been any mucosal rupture (biopsy or snare resection) or not. As a group, those who attended in 1983 and were informed that they had polyps tended to improve their smoking habits more than those informed that they had no polyps. Similarly, and in spite of more people giving up smoking, the group with polyps had a trend towards a smaller increase in BMI (+0.7 (95% CI 0.2-1.1)) than the polyp-free group (+1.2 (95% CI 0.9-1.6)) (P = 0.07). The observations suggest that flexible sigmoidoscopy screening may face an educational challenge to avoid unfavourable changes in the lifestyle of screenees, an effect that may more than outweigh the beneficial effect of screening.

Drinking water and the prevalence of colorectal adenomas: an epidemiologic study in Telemark, Norway.

This study was based on an endoscopic screening study for detection of polyps in the rectum and sigmoid colon in a randomized, normal population sample of 400 individuals aged 50-59 years. Family disposition for cancer and indicators of lifestyle (including dietary registration) were recorded. The 310 individuals received domestic drinking water from one out of four public water supplies. The participants were categorized according to the water supply connected to their house of residence. Drinking water was analysed monthly during 2 years for chloroform, total organic carbon, colour index, calcium, magnesium and chlorine. The overall prevalence of colorectal polyps was significantly higher in residents receiving chlorinated water with a high organic content when compared with recipients of water with a low organic content. There was no association between polyp prevalence and chloroform concentration in the drinking water. Multivariate analysis revealed that age, male sex, high BMI, smoking, few stools per week, high protein intake and low intake of fibre, iron and cruciferous vegetables were far more important for the presence of polyps than the total organic content in chlorinated drinking water.

Assessment of Mucosal Inflammation and Circulation in Response to Probiotics in Patients Operated with Ileal Pouch Anal Anastomosis for Ulcerative Colitis.

BACKGROUND: Pouchitis is a common and troublesome condition, and a disturbed microbiological flora and mucosal blood flow in the pouch have been suggested as possible causes. Laser Doppler flowmetry (LDF) has been used successfully to measure gastric and colonic mucosal perfusion in humans. The aim of this study was to evaluate the effect of intervention with probiotics on ileal pouch inflammation and perfusion in the pouch, assessed by endoscopy, histology, fecal calprotectin and LDF. METHODS: A fermented milk product (Cultura; 500 ml) containing live lactobacilli (La-5) and bifidobacteria (Bb-12) was given daily for 4 weeks to 10 patients operated with ileal-pouch-anal anastomosis (IPAA) for ulcerative colitis (UC). Mucosal perfusion was measured with LDF and the degree of inflammation was examined at predefined levels of the distal bowel by endoscopy and histology. Stool samples were cultured for lactobacilli and bifidobacteria and calprotectin were measured before and after intervention. RESULTS: The LDF measurements were reproducible in the pelvic pouch at each of the predefined levels, but did not change after intervention. The mucosal perfusion was reduced in the distal compared to the proximal part of the pouch. Calprotectin levels did not change significantly after intervention. The median endoscopic score for inflammation was significantly reduced by 50% after intervention, whereas the histological score did not change significantly. CONCLUSION: The results suggest that probiotics primarily act superficially, with change of gross appearance of the mucosa at endoscopy, but without significant effect on histological picture, mucosal perfusion or faecal calprotectin, during a relatively short period of 4 weeks.

Results from two repeated 5 day dietary records with a 1 y interval among patients with colorectal polyps.

BACKGROUND: Dietary factors are known to be associated with initiation and development of colorectal cancer (CRC), and also with CRC's major precursor, the colorectal polyp. In long-term intervention studies on colorectal polyps, dietary changes may therefore affect potential effects of the study intervention. OBJECTIVE: To examine potential dietary changes among polyp-patients randomly selected from a 3 y intervention study after 1 y. DESIGN AND SUBJECTS: Of 116 polyp-bearing out-patients (50% men), aged 50-76 y, who participated in the double-blind 3 y placebo-controlled endoscopic follow-up and intervention study against growth and recurrence of polyps, 30 patients were randomised (strata: sex, age and polyp size) to perform a repeated 5 day dietary record by weighing after 1 y. The patients received a daily mixture of vitamin C (150 mg), alpha-tocopherol (75 mg), beta-carotene (15 mg), selenium (101 microg) and calcium (1.6 g) or placebo (lactose) for a period of 3 y with annual colonoscopic examinations and polyps size measurements to test if the mixture was able to reduce polyp growth and recurrence. Polyps of >9 mm were removed, whereas the remainders and new discoveries of polyps <9 mm were left in situ until the end of the study. RESULTS: Twenty-nine patients agreed to perform the repeated 5 day dietary record, and 86% performed the second record within 48-58 weeks after the first record. The results showed that, with the exception of vitamin D, milk and milk products, no significant differences were found between the two records. The median value of the Spearman's correlation coefficient for energy and energy-yielding nutrients was 0.66, for vitamins and minerals 0.58, and for foods 0.58. Individual differences between the records were found for most variables, but most of these were negligible. CONCLUSION: After 1 y, no major dietary changes were found which could be associated with a changed susceptibly for malignancy, and thereby affect potential effects of the study intervention. We may thus suggest that a potential changed susceptibility towards growth and recurrence of polyps, is due to the specific intervention, and not due to other major dietary changes.

Food intolerance and psychosomatic experience.

The mechanism behind food intolerance is regarded as one of the greatest enigmas in modern medicine. Its multidisciplinary modalities, sharing properties with immunologic, environmental and psychosomatic reaction patterns, make the grouping and individual approach rather complex in regard to classification of disease, diagnosis, and therapy. In this presentation, emphasis is placed on emerging knowledge about immunologic reactions in the bowel and blood circulation as a balance against the evidence for psychosomatic reactions. As a basis for discussion, the psychosomatic experience of patients with food intolerance is illustrated by a brief presentation of three studies. The first was cross-sectional. The second was prospective and controlled. The third was a double-blind placebo-controlled study using provocation with an active substance in comparison with a placebo. Both the patients and referents were characterized by interviews and scoring systems based on questionnaires. When either combined or kept separately, the results of these studies suggest a correlation between somatic and neuropsychiatric symptoms and emotional disturbances. It also seems that patients identifying themselves as sensitive to food and chemicals have higher scores for depression, anxiety, shyness, and defensiveness. On the other hand, in 62% of the cases, there was agreement between diet history and provocation. The next-of-kin of the food intolerant subjects also had various diseases more frequently, increased immunoglobulin E levels, and a higher prevalence of allergy and infectious diseases. For the same patients, major distress or trauma during childhood, as well as undifferentiated somatoform disorders, were common. In conclusion, both somatic symptomatology and self-reported psychological disturbances can be regarded as rather weak documentations. The experience within these fields today may, however, seem promising for further research. One should then emphasize the importance of the nature of exposure and the nature of disposition, represented by immunologic or psychological mechanisms, or a combination of both. Future studies should be aimed at classifying patients into subgroups through the use of improved diagnostic and clinical methods, assessment of organ sensitivity, and immunologic and psychological tests.

Symptoms and manifestations of food intolerance.

Food intolerance (FI) is a less defined and a more heterogenous diagnosis than food allergy. The characterization of symptoms during trials, and relationship to time, meal and events such as provocation, are cornerstones of the diagnosis, which is hampered by the complexity of mechanisms and individual conceptuality of illness. Most cases of food intolerance are seen in general practice or do not seek health care. Nevertheless, a substantial number of patients are refered to hospitals. Variation in frequency of diagnosis may be caused by differences in health care levels, dietary habits and age. General symptoms may occur alone or in combination with symptoms from one or more target organs. Even the double blind provocation may give both false positive and false negative results. Repeat examinations, open provocations or verification by a subsequent follow up on an elimination diet may be needed. Many patients are satisfied with an elimination diet given after provocation. The primary objective, however, is to exclude specific organic or psychosocial explanations, in a multidisciplinary approach to these patients.

Enzyme activity and protein patterns as premalignant markers in mucosal biopsy specimens from the large intestine.

Total enzyme activity of glucose-6-phosphate dehydrogenase (G6PD) and lactate dehydrogenase (LD) was measured in homogenates of resected biopsy specimens and in endoscopic biopsy specimens. LD isoenzyme patterns were scanned by a laser technique after agarose gel electrophoresis. Examinations were performed in homogenates of premalignant lesions such as ulcerative colitis and adenomas of the colon, with normal mucosa and carcinomas as control material. Additionally, two-dimensional electrophoretic protein patterns were compared for normal mucosa, adenomas, and carcinomas of the large intestine. The mean activity of both G6PD and LD was highest in the presence of dysplasia; however, only G6PD activity seemed independent of inflammatory changes. The percentage of LD isoenzyme M monomers was significantly higher in homogenates of specimens with dysplastic changes than in specimens with only inflammatory changes. A positive correlation was found between total LD isoenzyme M monomers and LD 5 monomers for the whole material and for each of the histological subgroups of ulcerative colitis. A positive correlation between total LD activity and the percentage of LD 5 monomers was seen only for dysplastic, adenomatous, and malignant tissues. The several hundred protein spots seen on two-dimensional protein maps showed that most of the spots were common for normal mucosa, adenomas, and carcinomas, but differences were also seen. Polyps and carcinomas had strikingly similar protein patterns, different from that of normal mucosa. The results of the two-dimensional protein electrophoresis lend further support to the hypothesis that polyps are precursors of carcinomas of the large intestine.(ABSTRACT TRUNCATED AT 250 WORDS)