Autonomic nervous system function predicts the inflammatory response over three years in newly diagnosed ulcerative colitis patients.

BACKGROUND: The autonomic nervous system (ANS) modulates intestinal inflammation in animal models. Human evidence confirming such modulating influence is limited. We aimed to investigate whether ANS function is associated with inflammatory parameters at disease onset, and whether it predicts the evolution of inflammation in patients with ulcerative colitis (UC). METHODS: We prospectively monitored 51 patients from onset of UC for 3 years. Upon remission of the onset flare, ANS activity was assessed by heart rate variability analysis and compared with healthy controls. Inflammatory parameters in blood, stool, and colonic biopsies obtained at onset and during follow-up visits were analyzed. Generalized linear models were used to test cross-sectional associations between ANS activity and inflammatory parameters at onset; linear mixed models were used to test whether ANS function at onset predicted the evolution of inflammation over the following 3 years. KEY RESULTS: Sympathovagal balance was different in UC patients compared to healthy controls, and cross-sectional associated with higher levels of systemic (erythrocyte sedimentation rate [ESR], CRP, TNF-α, IFN-γ) and mucosal inflammation (interleukin-8, IFN-γ) at onset. Conversely, a negative cross-sectional association with parasympathetic activity was found for ESR & TNF-α. Longitudinally, parasympathetic activity at onset predicted systemic (ESR, WBC), but not mucosal inflammation during follow-up. CONCLUSIONS & INFERENCES: This study further strengthens the association between the ANS system and intestinal inflammation previously found in animal models and recently in patients with inflammatory bowel disease. These results may have important implications for the pathogenesis and treatment of UC.

Cultured blood T-cell responses predict anti-TNF therapy response in patients with ulcerative colitis.

BACKGROUND: Anti-tumour necrosis factor (TNF) therapy is used for treatment of ulcerative colitis (UC). As approximately 30% of patients with UC do not benefit from the treatment, it is of clinical interest to identify biomarkers of response before therapy is initiated. AIM: To identify prognostic biomarkers of anti-TNF therapy response in anti-TNF therapy-naïve patients with UC. METHODS: Peripheral blood cells were obtained from 56 patients with UC before therapy started. Thirty-four patients were included in an exploratory cohort and 22 patients in a validation cohort. Blood cells were stimulated in vitro with influenza vaccine with and without anti-TNF. T-cell surface receptor expression and cytokine release were determined (in total 17 variables). Treatment response was evaluated using the Mayo score 12-14 weeks after the first infusion. RESULTS: In the exploratory cohort, blood cells from the patients showed stronger anti-TNF-dependent suppression of T-cell surface receptor expression and cytokine secretion among therapy responders than nonresponders. In particular, anti-TNF suppressed the expression of CD25 on T cells and secretion of interleukin 5, to a higher degree in responders than in nonresponders. These variables were used to a create model to predict therapy outcome, which was confirmed in the validation cohort. Correct classification of future therapy response was achieved in 91% of the cases in the validation cohort. CONCLUSION: The effects of anti-TNF on cultured blood T cells, obtained before therapy started, predict treatment outcome in patients with UC.

The unsaponifiable fraction of extra virgin olive oil promotes apoptosis and attenuates activation and homing properties of T cells from patients with inflammatory bowel disease.

The unsaponifiable fraction (UF) of extra virgin olive oil (EVOO) possesses anti-inflammatory properties and exerts preventative effects in murine models of inflammatory bowel disease (IBD). The present study was designed to determine the in vitro effects of UF on blood and intestinal T cells from IBD patients and healthy subjects. The T cell phenotype was investigated by flow cytometry and cytokine secretion was determined by ELISA. The presence of UF of EVOO promoted apoptosis and attenuated activation of intestinal and blood T cells isolated from IBD patients, decreasing the frequency of CD69(+) and CD25(+) T cells and, also, the secretion of IFN-γ. Moreover, UF reduced the expression of the gut homing receptor integrin ß7 on blood T cells from IBD patients. In conclusion, UF modulates the activity and the gut homing capacity of T cells, and might therefore be considered as a dietary complement with an anti-inflammatory role in IBD patients.

Characterization of IBS-like symptoms in patients with ulcerative colitis in clinical remission.

BACKGROUND: Gastrointestinal symptoms compatible with Irritable Bowel Syndrome (IBS) are common in patients with inflammatory bowel disease. It has been suggested that these symptoms are a reflection of occult inflammation rather than coexisting IBS. The aim of this study was to characterize IBS-like symptoms in patients with Ulcerative Colitis (UC) in clinical remission by assessing inflammatory markers, psychological symptoms, and quality of life. METHODS: Ninety-four patients with new onset of UC were followed prospectively during 3 years with yearly follow-up visits. The patients completed self-administrated questionnaires. Fecal calprotectin was used as an inflammatory biomarker. Remission was defined as a total Mayo-score ≤2 and an endoscopic subscore ≤1, with no relapse during the 3-month period prior to visit. KEY RESULTS: The prevalence of patients that fulfilled Rome II criteria for IBS among UC patients in remission was 11% at visit 1, 23% at visit 2, and 17% at visit 3. When comparing UC patients in remission with and without IBS-like symptom, patients with IBS-like symptoms had more severe gastrointestinal symptoms, tendencies toward more severe psychological symptoms and reduced levels of quality of life, but the calprotectin levels did not differ between the two groups. CONCLUSIONS & INFERENCES: IBS-like symptoms are common in patients with UC in clinical remission and these fluctuate over time. The symptoms are associated with poor psychological well-being and reduced quality of life, and do not seem to be a reflection of low-grade inflammatory activity.

Infliximab inhibits activation and effector functions of peripheral blood T cells in vitro from patients with clinically active ulcerative colitis.

Many patients with inflammatory bowel disease (IBD) are undergoing therapy with infliximab, an antibody specific for TNF. However, the exact mechanisms of action of infliximab are not completely understood. The aim of this study was to determine the in vitro effects of infliximab on blood T cells derived from anti-TNF therapy-naïve ulcerative colitis (UC) patients with clinically active disease. Peripheral blood mononuclear cells were stimulated polyclonally or by antigen in the presence or absence of infliximab. The T cell phenotype was investigated by flow cytometry, cytokine secretion was determined by ELISA, and cell proliferation was determined by thymidine assay or CFSE dye. Presence of infliximab resulted in reduced expression of CD25 in CD4(+) and CD8(+) T cell populations and inhibited secretion of IFN-γ, IL-13, IL-17A, TNF as well as granzyme A. Infliximab also suppressed CD4(+) and CD8(+) T cell proliferation. These effects of infliximab were recorded both in T cells activated by polyclonal and antigen-specific stimulation. The effects of infliximab on T cell apoptosis and induction of FOXP3(+) CD4(+) T regulatory cells were ambiguous and depended on the originating cellular source and/or the stimulation mode and strength. In conclusion, infliximab is able to reduce T cell activation as measured by CD25, proliferation and cytokine secretion in vitro from UC patients with clinically active disease. These data suggest that suppression of T cell activity may be important for infliximab-mediated disease remission in patients with UC.

Clinical trial: the effects of a fermented milk containing three probiotic bacteria in patients with irritable bowel syndrome - a randomized, double-blind, controlled study.

BACKGROUND: The effects of probiotic bacteria in IBS remain controversial. AIM: To study the effects of a probiotic product on IBS symptoms. METHODS: We randomized 74 IBS patients to receive 8 weeks of daily treatment with 400 mL milk fermented with the yoghurt bacteria and containing Lactobacillus paracasei, ssp. paracasei F19, Lactobacillus acidophilus La5 and Bifidobacterium lactis Bb12 (Cultura; active) or acidified milk without these bacteria (control). The primary endpoint was the proportion of subjects reporting adequate relief of their IBS symptoms at least 50% of the weeks. IBS symptom severity, psychological symptoms and quality of life were assessed. RESULTS: The proportion of responders was 38% (14/37 patients) in the active group and 27% (10/37 patients) in the control group (P = 0.3). IBS symptom severity improved significantly in both groups during the treatment period. This change was greater in the active group during the first 2 weeks, but thereafter, no significant group differences were seen. CONCLUSIONS: We could not detect a clearly positive effect of fermented milk containing three probiotic bacteria on GI symptoms in IBS patients compared with the control treatment. However, a trend towards a more favourable effect during the first weeks was seen in the active group.

B-cell activation in patients with irritable bowel syndrome (IBS).

Patients with irritable bowel syndrome (IBS) may have a low grade immune activation. However, little is known about the properties of B cells of IBS patients. We therefore investigated activation level and antigen presenting phenotype of blood B cells of IBS patients. We also examined B-cell responses to lipopolysaccharide (LPS) and probiotic bacteria. Blood samples were obtained from 74 IBS patients and 30 healthy subjects. Peripheral blood mononuclear cells were isolated and stimulated with LPS or an UV-light inactivated bacterial cocktail consisting of the probiotic Gram-positive strains; Lactobacillus paracasei ssp. paracasei 19, Lactobacillus acidophilus La5, Bifidobacterium lactis B612. The phenotype of CD19(+) B cells was investigated by flow cytometry before and after 72 h cell culture. Furthermore, IBS symptom severity was assessed. B cells isolated from blood of IBS patients displayed an amplified activation level as demonstrated by increased cell surface expression of IgG, and also the costimulatory molecules CD80 and CD86. Expression of antigen presenting HLA-DR and costimulatory molecule CD40 on B cells was, however comparable in IBS patients and controls. B cells of IBS patients displayed an impaired ability to increase expression of CD80, but not CD86, in response to both LPS as well as probiotic bacteria stimulations. To conclude, blood B cells of IBS patients have an increased activation level. Bacterial component induced expression of the costimulatory molecule CD80, regarded as important for tolerance induction, is impaired. These data suggest that B-cell antigen presentation in IBS patients is associated with altered capacity of providing costimulation to T cells.

Phenotypic and genotypic characterization of coagulase-negative staphylococci isolated in blood cultures from patients with haematological malignancies.

Coagulase-negative staphylococci are the predominant aetiological agents in bacteraemic patients hospitalized for haematological malignancies. The aim of this study was to determine whether differences exist in the prevalence of icaAB genes and in the phenotypic and/or genotypic pattern between blood isolates of coagulase-negative staphylococci, interpreted as representing true bacteraemia, and contaminant isolates from patients with haematological malignancies. Eighty-two isolates representing true bacteraemia and 47 contaminant isolates were found among 76 patients. The most prevalent species in both groups of patients was Staphylococcus epidermidis (n=103; 80%). Biochemical typing using the Phene Plate system and genotyping using pulsed-field gel electrophoresis showed a tendency towards a more homogeneous pattern among isolates causing true bacteraemia compared with contaminant isolates. Two major genotypic groups of S. epidermidis were found in both the true bacteraemia group and the contaminant group, with concordant pulsotypes found as well. These groups may comprise isolates carrying specific virulence factors, but the prevalence of the icaAB genes did not differ between the true bacteraemia group and the contaminant group. No significant difference was seen between the two study groups regarding clinical symptoms or complications, use of central venous catheter, and levels of absolute neutrophil count or C-reactive protein.

Delay in gastric emptying in patients with chronic renal failure.

BACKGROUND: Gastrointestinal (GI) symptoms are common in patients with chronic renal failure (CRF). Delayed gastric emptying might be a possible pathophysiological mechanism. The aims of this study were to evaluate gastric emptying in patients with CRF and to correlate the findings with GI symptoms and evaluate the impact of Helicobacter pylori infection in CRF patients on gastric emptying. METHODS: Thirty-nine patients with CRF (17 F, 22 M) were compared with 131 healthy subjects (74 F, 57 M). A standardized breakfast was given with 20 spherical, radiopaque markers (ROMs). The emptying was followed by fluoroscopy after 4, 5 and 6 h. Gastric emptying was assessed by calculating the individual mean percentual gastric retention of markers, 4 to 6 h after the meal. The perceived severity of GI symptoms was assessed with a validated questionnaire. Because of gender differences in gastric emptying, men and women were compared separately and a percentile of 95 was chosen as the upper reference value. H. pylori infection was assessed using a serological method. RESULTS: Delayed gastric emptying was found in 14 out of 39 (36%) of the CRF patients. There was no relationship between delayed gastric emptying and age, GI symptoms, H. pylori infection or underlying renal disease. However, a higher proportion of patients in peritoneal dialysis demonstrated delayed gastric emptying compared with predialytic patients (6 of 9 versus 2 of 13, P = 0.026). Men with CRF had a higher gastric retention compared with healthy men (16.6 (0-63.3)% versus 0 (0-2.1)%, P < 0.0001), and 10 men with CRF had delayed gastric emptying (P < 0.0001). There was no significant difference in mean gastric retention between women with CRF and healthy women (13.3 (0-55.4)% versus 10.8 (0-30.0)%, P = 0.93), but 4 women with CRF had delayed gastric emptying (P = 0.02). Eighteen of the CRF patients had GI symptoms (6 F, 12 M) and 21 were asymptomatic (11 F, 10 M). There was no difference in mean gastric retention in patients with CRF with and without GI symptoms (M: 13.3 (0-55.0)% versus 47.5 (5.0-65.0)%, P = 0.51, F: 16.6 (0-63.3)% versus 13.3 (0-59.2)%, P = 0.96). Gastric emptying in CRF patients with and without H. pylori infection showed no difference. CONCLUSIONS: Delayed gastric emptying is common in patients with chronic renal failure, particularly in men. The delay was not associated with the presence of GI symptoms, underlying renal disease or H. pylori infection. However, the dialytic status might have an impact on gastric emptying in patients with CRF.

Overuse of acid suppressive therapy in hospitalised patients with pulmonary diseases.

OBJECTIVES: Overuse of acid suppressive therapy (AST) has been reported in hospitalised patients, but the use in specific patient categories is unexplored. We assessed the use of and indication for AST and upper endoscopic investigations in hospitalised patients on a pulmonary ward compared with patients on other wards. METHODS: 301 patients were enrolled in the study. 162 were hospitalised on a pulmonary ward with a control group consisting of 139 from both a surgical and general internal medicine ward. Adequate indications for AST were those strongly supported by medical literature. RESULTS: Among the 301 patients enrolled, 132 (44%) used AST. 78 (59%) had no adequate indication for AST. On the pulmonary ward 79 (49%) patients used AST, compared to only 10 (20%) on the internal medicine ward (P < 0.05). On the pulmonary ward 68% of the patients had no adequate indication for AST, which was more common than inappropriate use of ASTon the control wards (P < 0.05). The most common inadequate indication for AST was peptic ulcer prophylaxis during corticoidsteroid therapy. CONCLUSION: In hospitalised patients a significant overuse of AST was observed, particularly among pulmonary patients. More adequate use of AST can contribute to substantial savings for the health-care system.

ATP dependent Ca2+ transport across basal membrane of human syncytiotrophoblast in pregnancies complicated by intrauterine growth restriction or diabetes.

Neonates born after pregnancies complicated by diabetes or intrauterine growth restriction (IUGR) have increased incidence of hypocalcaemia. Furthermore, IUGR is associated with reduced bone mineralization in infancy and osteoporosis in adult life. We tested the hypothesis that placental calcium transport is altered in these pregnancy complications. Transport of calcium into syncytiotrophoblast basal plasma membrane (BM) vesicles was studied by rapid filtration and protein expression of Ca(2+) ATPase by Western blot. In IUGR Ca(2+) ATPase activity was increased by 48 per cent (n=13; P< 0.05) whereas protein expression was 15 per cent lower (n=13; P< 0.05) than in controls (n=16). Basal membrane ATP dependent calcium transport was unaltered in gestational diabetes (GDM) but increased by 54 per cent in insulin dependent diabetes (IDDM) compared to controls (P< 0.05; n =14). Diabetes did not affect Ca(2+) ATPase expression in BM. We have previously shown that the mid-molecular fragment of parathyroid hormone related peptide (PTHrP midmolecule) stimulates BM Ca(2+) ATPase in vitro. PTHrP midmolecule concentrations in umbilical cord plasma were measured using radioimmunoassay. The concentrations in umbilical cord plasma were increased in IUGR, but unaltered in diabetes. In conclusion, placental calcium pump is activated in IUGR and IDDM, which may be secondary to increased foetal calcium demand. We speculate that PTHrP midmolecule may be one mechanism for activating BM Ca(2+) ATPase in IUGR.

Parathyroid hormone-related peptide (38-94) amide stimulates ATP-dependent calcium transport in the Basal plasma membrane of the human syncytiotrophoblast.

The final step in the maternal-fetal transfer of calcium in the placenta involves transport against a concentration gradient across the syncytiotrophoblast basal plasma membrane (BM). Based on animal studies, it has been proposed that parathyroid hormone-related peptide (PTHrP) plays a major role in maintaining the maternal-fetal concentration gradient of calcium. In this study, we tested the hypothesis that a highly conserved mid-region fragment (38-94) of PTHrP directly affects the ATP-dependent calcium transport across BM isolated from full-term human placentas. PTHrP (38-94) stimulated ATP-dependent calcium transport at a concentration within the physiological range (5 pg/ml) and the effect (10-38% increase) was concentration dependent over the range 5 pg/ml to 5 ng/ml (n=8; P<0.05). In contrast, PTH, PTHrP (1-34), PTHrP (67-86) and calcitonin increased BM calcium transport only at concentrations much higher than physiological. The increased calcium uptake was inhibited by the protein kinase C (PKC) inhibitor chelerythrine (n=6; P<0.05). In addition, PTHrP (38-94) increased inositol trisphosphate (IP(3)) production and PKC phosphorylation in human placental BM (n=12; P<0.05). Our data indicate that PTHrP (38-94) stimulates Ca(2+)ATPase in the human syncytiotrophoblast BM vesicles by activating the IP(3)-DAG-PKC pathway. We suggest that PTHrP (38-94) is important in maintaining the calcium concentration gradient across the placental barrier in the human.

Impact of sex and psychological factors on the water loading test in functional dyspepsia.

BACKGROUND: The pathogenesis of upper abdominal symptoms in patients with functional dyspepsia (FD) is still unclear. The water loading test (WLT) is a new method for evaluation of gastric function. Our aim was to determine the impact of sex, age and psychological factors on the results of WLT in FD patients, FD-subgroups and healthy controls (HCs), and to evaluate the safety of the test. METHODS: Fifty-six HCs and 35 consecutive patients with FD drank mineral water (100 ml/min) until intolerable. Serum samples for sodium, potassium and creatinine was taken before and after drinking. Water quantity was registered and symptoms were assessed after maximal water intake and 30 min later using a VAS scale. Participants also completed questionnaires measuring psychological general well-being (PGWB) and gastrointestinal symptoms (GSRS and Mearin score). RESULTS: Healthy men drank more than healthy women, 2350 +/- 105 ml versus 1860 +/- 100 ml (P < 0.01), and the same gender difference was noted in FD patients, 1770 +/- 115 ml versus 1180 +/- 110 ml (P < 0.01). Maximal water intake was significantly higher in HC than in FD patients, both in males (P < 0.001) and females (P < 0.0001). Age had no impact on drinking capacity. FD patients had more symptoms 30 min after maximal water intake than HCs. Serum sodium decreased from 141 +/- 0.3 mmol/l to 138 +/- 0.5 mmol/l. Two of the assessed psychological factors, general health and depressed mood, correlated with water intake in FD patients (Rho = 0.47, P < 0.01 respectively Rho = 0.41, P < 0.05). CONCLUSION: WLT is a useful, simple and safe test for evaluating symptoms in FD patients. Sex, but not age affects the results of the WLT. Furthermore, psychological factors must also be taken into consideration when interpreting the WLT.

ATP-dependent Ca2+ transport is up-regulated during third trimester in human syncytiotrophoblast basal membranes.

In late gestation, Ca2+ transport across the human placenta must increase in response to the demands of accelerating bone mineralization of the fetus. This is an ATP-dependent transport against a concentration gradient across the basal or the fetal-facing plasma membrane of the syncytiotrophoblast. The aims of the present study were to determine the relationship between ATP-dependent Ca2+ transport and gestational age in the third trimester and to identify the specific isoforms of plasma membrane Ca2+ ATPase (PMCA) present in human syncytiotrophoblast. Basal membrane vesicles were isolated from normal placentas and from placentas obtained from preterm deliveries with no other complications (32-37 wk of gestation). We studied the uptake of 45Ca2+ into basal membrane vesicles in the absence and presence of ATP by using rapid filtration techniques. Western blot was used to assess the protein expression of the PMCA isoforms 1-4. Isoforms 1 and 4 of PMCA were identified in basal membrane of human placenta. The ATP-dependent Ca2+ transport increased linearly during the third trimester (r = 0.571, p = 0.0015, n = 28). However, PMCA protein expression was unaltered during the same period of gestation. Our results show that PMCA in the fetal-facing plasma membrane of the human syncytiotrophoblast is markedly activated toward the end of pregnancy. We suggest that these changes are critical in supplying the rapidly growing fetus with sufficient Ca2+ for bone mineralization.

Molecular markers for UV-B stress in plants: alteration of the expression of four classes of genes in Pisum sativum and the formation of high molecular mass RNA adducts.

Sixteen ultraviolet-B radiation-regulated pea genes were identified. Functionally, the corresponding proteins were divided into four groups. (i) Chloroplast-localized proteins. Genes for these proteins were down-regulated, underlining the deleterious effects of UV-B on this organelle. A novel down-regulated photosystem I light-harvesting chlorophyll a/b-binding protein gene (PsLhcA4), was cloned and sequenced. (ii) Protein turnover enzymes. Levels of mature mRNAs for the PU1 and PsUBC4 genes, encoding proteins of the ubiquitin protein degradation pathway, were up- and down-regulated, respectively, implying alteration of plant cell protein content by changes in both gene expression and protein degradation. (iii) Proteins involved in intracellular signalling. Expression of genes for small GTPases, rab and rho homologues, were altered. (iv) Phenylpropanoid or flavonoid biosynthesis. Expression of three genes encoding enzymes in these pathways were up-regulated and one of them, the novel PsC450R1, was cloned and sequenced. Moreover, unexpected high molecular mass psbA RNA adducts were found to appear after UV-B exposure. In addition, a large increase in corresponding high molecular mass adducts were also found for PsLhcA4, and PsUBC4 mRNA and 23S rRNA. These RNA species do not contain protein and probably appear due to cross-linking of two or more RNA molecules, or are the result of UV-B-induced failure of transcription termination.