Serum 1,3-beta-D-glucan as a noninvasive test to predict histologic activity in patients with inflammatory bowel disease.

BACKGROUND: 1,3-beta-D-glucan (BG) is a ubiquitous cell wall component of gut micro-organisms. We hypothesized that the serum levels of BG could reflect active intestinal inflammation in patients with inflammatory bowel disease. AIM: To determine whether the serum BG concentrations correlate with intestinal inflammation. METHODS: A prospective observational study was performed in a tertiary referral center, from 2016 to 2019, in which serum BG was determined in 115 patients with Crohn's disease (CD), 51 with ulcerative colitis (UC), and 82 controls using a photometric detection kit. Inflammatory activity was determined by ileocolonoscopy, histopathology, magnetic resonance enterography, and biomarkers, including fecal calprotectin (FC), C-reactive protein, and a panel of cytokines. The ability of BG to detect active vs inactive disease was assessed using the area under the receiver operating characteristic curve. In subgroup analysis, serial BG was used to assess the response to therapeutic interventions. RESULTS: The serum BG levels were higher in CD patients than in controls (P = 0.0001). The BG levels paralleled the endoscopic activity in CD patients and histologic activity and combined endoscopic and histologic activity in both CD and UC patients. The area under the curve (AUC) in receiver operating characteristic analysis to predict endoscopic activity was 0.694 [95% confidence interval (CI): 0.60-0.79; P = 0.001] in CD, and 0.662 (95%CI: 0.51-0.81; P = 0.066) in UC patients. The AUC in receiver operating characteristic analysis to predict histologic activity was 0.860 (95%CI: 0.77-0.95; P < 0.001) in CD, and 0.786 (95%CI: 0.57-0.99; P = 0.015) in UC patients. The cut-off values of BG for both endoscopic and histologic activity were 60 µg/mL in CD, and 40 µg/mL in UC patients. Performance analysis showed that the results based on BG of 40 and 60 µg/mL were more specific for predicting endoscopic activity (71.8% and 87.2% for CD; and 87.5% and 87.5% for UC, respectively) than FC (53.3% and 66.7% for CD; and 20% and 80% for UC, respectively); and also histologic activity (60.5% and 76.3% for CD; and 90.0% and 95.0% for UC, respectively) than FC (41.7% and 50.0% for CD; and 25% and 50% for UC, respectively). Regarding the clinical, endoscopic, and histologic activities, the BG levels were reduced following therapeutic intervention in patients with CD (P < 0.0001) and UC (P = 0.003). Compared with endoscopic (AUC: 0.693; P = 0.002) and histologic (AUC: 0.868; P < 0.001) activity, no significant correlation was found between serum BG and transmural healing based on magnetic resonance enterography (AUC: 0.576; P = 0.192). Positive correlations were detected between BG and IL-17 in the CD (r: 0.737; P = 0.001) and the UC group (r: 0.574; P = 0.005), and between BG and interferon-gamma in the CD group (r: 0.597; P = 0.015). CONCLUSION: Serum BG may represent an important novel noninvasive approach for detecting mucosal inflammation and therapeutically monitoring inflammatory bowel diseases, particularly in CD.

Prevalence, Indirect Costs, and Risk Factors for Work Disability in Patients with Crohn's Disease at a Tertiary Care Center in Rio de Janeiro.

BACKGROUND AND AIMS: Crohn's disease (CD) can lead to work disability with social and economic impacts worldwide. In Brazil, where its prevalence is increasing, we assessed the indirect costs, prevalence, and risk factors for work disability in the state of Rio de Janeiro and in a tertiary care referral center of the state. METHODS: Data were retrieved from the database of the Single System of Social Security Benefits Information, with a cross-check for aid pension and disability retirement. A subanalysis was performed with CD patients followed up at the tertiary care referral center using a prospective CD database, including clinical variables assessed as possible risk factors for work disability. RESULTS: From 2010 to 2018, the estimated prevalence of CD was 26.05 per 100,000 inhabitants, while the associated work disability was 16.6%, with indirect costs of US$ 8,562,195.86. Permanent disability occurred more frequently in those aged 40 to 49 years. In the referral center, the prevalence of work disability was 16.7%, with a mean interval of 3 years between diagnosis and the first benefit. Risk factors for absence from work were predominantly abdominal surgery, anovaginal fistulas, disease duration, and the A2 profile of the Montreal classification. CONCLUSIONS: In Rio de Janeiro, work disability affects one-sixth of CD patients, and risk factors are associated with disease duration and complications. In the context of increasing prevalence, as this disability compromises young patients after a relatively short period of disease, the socioeconomic burden of CD is expected to increase in the future.

Geographic distribution and time trends of esophageal cancer in Brazil from 2005 to 2015.

The purpose of the present study was to investigate the geographical distribution and time trends of the incidence and lethality of esophageal cancer (EC) in Brazil. The present study conducted an ecological study of EC using records from January 2005 to December 2015 in the Health Informatics Department of the Brazilian Ministry of Health (DATASUS) registry. In addition to demographical data on the population, EC incidence and lethality rates were estimated from hospitalizations and in-hospital mortalities and were adjusted by total available hospital beds. The adjusted EC rates per 100,000 increased from 9.1 in 2005 to 12.1 in 2015. The prevalence among males increased from 69 to 78%, while the female rates remained stable over the same period. Although EC was the most common in South and Southeast Brazil, the rates increased proportionately more in the other regions of the country, especially among males. Geographical analysis revealed higher rates of EC in more urbanized areas, with a coast-to-inland gradient. While rates increased in people older than 50 years, they decreased among people below this age. However, the lethality rates remained stable and high during the study period, overlapping with hospital admission rates. The recent increasing trend in the EC incidence, with shifts from the south towards the north and from more urbanized towards rural areas, suggests that environmental factors are crucial in EC pathogenesis. The concentration of EC in South Brazil may reflect the presence of major environmental factors in association with a possible genetic predisposition. The unchanging high mortality associated with EC in the rapidly aging population suggests that EC will continue to impose a significant social and economic burden in the future.

Abdominal ultrasonography with color Doppler analysis in the assessment of ileal Crohn's disease: comparison with magnetic resonance enterography.

BACKGROUND/AIMS: Consistently defining disease activity remains a critical challenge in the follow-up of patients with Crohn's disease (CD). We investigated the potential applicability of abdominal ultrasonography with color Doppler (USCD) analysis for the detection of morphological alterations and inflammatory activity in CD. METHODS: Forty-three patients with CD ileitis/ileocolitis were evaluated using USCD analysis with measurements obtained on the terminal ileum and right colon. Sonographic parameters included wall thickening, stricture, hyperemia, presence of intra-abdominal mass, and fistulas. Patients were evaluated for the clinical activity (Harvey-Bradshaw Index [HBI]), fecal calprotectin (FC) and C-reactive protein (CRP). The USCD performance was assessed using magnetic resonance enterography (MRE) as a criterion standard. RESULTS: Most measurements obtained with USCD matched the data generated with MRE; however, the agreement improved in clinically active patients where sensitivity, positive predictive value, and accuracy were >80%, considering wall thickening and hyperemia. Complications such as intestinal wall thickening, stricture formation, and hyperemia, were detected in the USCD analysis with moderate agreement with MRE. The best agreement with the USCD analysis was obtained in regard to FC, where the sensitivity, positive predictive value, and accuracy were >70%. The overall performance of USCD was superior to that of HBI, FC and CRP levels, particularly when considering thickening, stricture, and hyperemia parameters. CONCLUSIONS: USCD represents a practical noninvasive and low-cost tool for evaluating patients with ileal or ileocolonic disease, particularly in clinically active CD. Therefore, USCD might become a useful asset in the follow-up of patients with CD.

Superiority of Interferon Gamma Assay Over Tuberculin Skin Test for Latent Tuberculosis in Inflammatory Bowel Disease Patients in Brazil.

BACKGROUND AND AIMS: To compare tuberculin skin test (TST) and interferon gamma release assay (IGRA) in the screening of LTBI among patients with inflammatory bowel disease (IBD) in an endemic area for tuberculosis, to evaluate the need for repeating tests during anti-TNFα, therapy, and to check whether the results may be affected by immunosuppression. METHODS: A cross-sectional study of 110 IBD patients and 64 controls was conducted in Rio de Janeiro, Brazil. The TST was administered after the Quantiferon(®)-TB Gold In-tube test was performed. RESULTS: TST and IGRA agreement was poor regarding diagnosis (kappa: control = 0.318; UC = 0.202; and CD = - 0.093), anti-TNFα therapy (kappa: with anti-TNFα = 0.150; w/o anti-TNFα = - 0.123), and immunosuppressive therapy (IST) (kappa: with IS = - 0.088; w/o IS = 0.146). Indeterminate IGRA was reported in four CD patients on IST. Follow-up tests after anti-TNFα identified conversion in 8.62% using TST and 20.0% using IGRA. Considering IGRA as a criterion standard, TST showed low sensitivity (19.05%) and positive predictive value (PPV) (21.05%). LTBI detection remarkably improved when IGRA was added to TST (sensitivity of 80.95% and PPV of 53.13%). Results were particularly relevant among CD patients where rates started from zero to reach sensitivity and PPV of more than 60%. CONCLUSION: IGRA alone was more effective to detect LTBI than TST alone and had an overall remarkable added value as an add-on sequential test, particularly in CD patients. While cost-effectiveness of these strategies remains to be evaluated, IGRA appears to be justified in CD prior to and during anti-TNFα therapy, where tuberculosis is endemic.

Changes in the Management of Patients with Crohn's Disease Based on Magnetic Resonance Enterography Patterns.

BACKGROUND AND AIMS: Magnetic resonance enterography (MRE) has become an important modality of radiological imaging in the evaluation of Crohn's disease (CD). The aim of this study was to investigate the impact of MRE in the assessment of disease activity and abdominal complications and in the making of therapeutic decisions for patients with CD. METHODS: In a cross-sectional retrospective study, we selected 74 patients with CD who underwent MRE and ileocolonoscopy with an interval between the two exams of up to 30 days between January 2011 and December 2017. We assessed the parameters of the images obtained by MRE and investigated the agreement with the level of disease activity and complications determined by a clinical evaluation, inflammatory biomarkers, and endoscopy, as well as the resulting changes in medical and surgical management. RESULTS: Changes in medical management were detected in 41.9% of patients. Significant changes in medical decisions were observed in individuals with a purely penetrating (P = .012) or a mixed (P = .024) MRE pattern. Patients with normal MRE patterns had a correlation with unchanged medical decisions (P = .001). There were statistically significant agreements between the absence of inflammatory criteria on MRE and remission according to the Harvey and Bradshaw index (HBI) (P = .037), the presence of inflammatory criteria on MRE and positive results for calprotectin (P = .005), and penetrating criteria on MRE and the scoring endoscopic system for Crohn's disease (SES-CD), indicating active disease (P = .048). Finally, there was significant agreement between the presence of fibrostenotic criteria and a long disease duration (P = .027). CONCLUSION: MRE discloses disease activity and complications not apparent with other modalities and results in changes in therapeutic decisions. In addition to being used for diagnosis, MRE should be routinely used in the follow-up of CD patients.

The socio-economic impact of work disability due to inflammatory bowel disease in Brazil.

BACKGROUND: Inflammatory bowel disease (IBD) might have economic and social impacts in Brazil, where its prevalence has increased recently. This study aimed to assess disability due to IBD in the Brazilian population and demographic factors potentially associated with absence from work. METHODS: Analysis was performed using the computerized Single System of Social Security Benefits Information, with a cross-check for aid pension and disability retirement, for Crohn's disease (CD) and ulcerative colitis (UC). Additional data were obtained from the platform, including the average values, benefit duration, age, gender and region of the country. RESULTS: Temporary disability occurred more frequently with UC, whereas permanent disability was more frequent with CD. Temporary disability affected more younger patients with CD than patients with UC. Temporary work absences due to UC and CD were greater in the South, and the lowest absence rates due to CD were noted in the North and Northeast. Absence from work was longer (extending for nearly a year) in patients with CD compared to those with UC. The rates of temporary and permanent disability were greater among women. Permanent disability rates were higher in the South (UC) and Southeast (CD). The value of benefits paid for IBD represented approximately 1% of all social security benefits. The benefits paid for CD were higher than for UC, whereas both tended to decrease from 2010 to 2014. CONCLUSIONS: In Brazil, IBD frequently causes disability for prolonged periods and contributes to early retirement. Reduction trends may reflect improvements in access to health care and medication. Vocational rehabilitation programs may positively impact social security and the patients' quality of life.

Disruption of the Hedgehog signaling pathway in inflammatory bowel disease fosters chronic intestinal inflammation.

Hedgehog (Hh) signaling is essential for intestinal homeostasis and has been associated with inflammation and tissue repair. We hypothesized that Hh signaling could affect the inflammatory process in inflammatory bowel disease (IBD). For this purpose, colon specimens from the inflamed and non-inflamed mucosa of 15 patients with Crohn's disease (CD), 15 with ulcerative colitis, and 15 controls were analyzed by immunohistochemistry and real-time PCR. The production and modulation of cytokines were measured by ELISA from culture explants. Apoptosis was assessed by TUNEL and caspase-3 activity assays. Chemotaxis was evaluated using a transwell system. Primary human intestinal and skin fibroblasts were used for analyzing migration and BrdU incorporation. Hh proteins were generally expressed at the superficial epithelium, and a marked reduction was observed in CD. In the lamina propria, Gli-1 predominantly co-localized with vimentin- and alpha-smooth muscle actin-positive cells, with lower levels observed in CD. In colon explants, Hh stimulation resulted in reduction, while blockade increased, TNF α, IL-17, and TGF ß levels. Apoptotic rates were higher in inflamed samples, and they increased after Hh blockade. Levels of Gli-1 mRNA were negatively correlated with caspase-3 activity. Hh blockade increased chemoattraction of monocytes. Primary fibroblasts incorporated more BrdU, but migrated less after Hh blockade. These results suggest that Hh signaling provides a negative feedback to the lamina propria, down-regulating inflammatory cytokines, and inhibiting leukocyte migration and fibroblast proliferation, while favoring fibroblast migration. Therefore, Hh signaling is strongly implicated in the pathogenesis of intestinal inflammation, and it may represent a novel therapeutic target for IBD.

Common NOD2/CARD15 and TLR4 Polymorphisms Are Associated with Crohn's Disease Phenotypes in Southeastern Brazilians.

AIM: To investigate whether variants in NOD2/CARD15 and TLR4 are associated with CD and ulcerative colitis (UC) in a genetically admixed population of Rio de Janeiro, where IBD has continued to rise. METHODS: We recruited 67 consecutive patients with CD, 61 patients with UC, and 86 healthy and ethnically matched individuals as controls. DNA was extracted from buccal brush samples and genotyped by PCR with restriction enzymes for G908R and L1007finsC NOD2/CARD15 single-nucleotide polymorphisms (SNPs) and for T399I and D299G TLR4 SNPs. Clinical data were registered for subsequent analysis with multivariate models. RESULTS: NOD2/CARD15 G908R and L1007finsC SNPs were found in one and three patients, respectively, with CD. NOD2/CARD15 G908R and L1007finsC SNPs were not found in any patients with UC, but were found in three and three controls, respectively. With regard to the TLR4 gene, no significant difference was detected among the groups. Overall, none of the SNPs investigated determined a differential risk for a specific diagnosis. Genotype-phenotype associations were found in only CD, where L1007finsC was associated with colonic localization; however, TLR4 T399I SNP was associated with male gender, and D299G SNP was associated with colonic involvement, chronic corticosteroid use, and the need for anti-TNF-alpha therapy. CONCLUSION: Variants of NOD2/CARD15 and TLR4 do not confer susceptibility to IBD, but appear to determine CD phenotypes in this southeastern Brazilian population.

Avaliação colonoscópica em pacientes com espondilite anquilosante / Colonoscopic evaluation in patients with ankylosing spondylitis

Introdução: Pacientes com espondilite anquilosante podem apresentar-se com lesões inflamatórias intestinais, e, por isso, deve ser definido o uso da colonoscopia para tais pacientes. Objetivos: Avaliar as alterações colonoscópicas intestinais macroscópicas e achados histopatológicos microscópicos de pacientes com espondilite anquilosante; correlacionar os achados colonoscópicos e histopatológicos; e estudar a relação dos achados histopatológicos com as manifestações extra-articulares da doença, HLA-B27, BASFI and BASDAI. Métodos: Este é um estudo transversal de 22 pacientes com espondilite anquilosante. Os pacientes passaram por uma avaliação clínica, BASDAI e BASFI, coleta de sangue para determinação de HLA-B27, e colonoscopia com biópsia de quarto segmentos intestinais (íleo terminal, cólon direito, cólon sigmoide e reto). Resultados: Resultados colonoscópicos anormais foram obtidos em 13 (59,1%) pacientes, e a principal anormalidade foi a presença de pólipos intestinais. Os grupos de resultados colonoscópicos normais e anormais (n = 9 e n = 13, respectivamente) foram homogêneos no que diz respeito à idade, BASFI, BASDAI, e variáveis categóricas, e o valor P não revelou diferença significativa entre grupos. Dos resultados histopatológicos, 81% tiveram uma biópsia anormal do íleo terminal, 90.9% tiveram uma biópsia anormal do cólon sigmoide, e a biópsia retal estava anormal em 86.4%. Os achados histopatológicos revelaram biópsias anormais em 81%, 90.9%, 90.9% e 86.4% para o íleo terminal, cólon direito, cólon sigmoide e reto, respectivamente. Os resultados histopatológicos não revelaram associação estatisticamente significativa com as manifestações ex...

Introduction: Patients with ankylosing spondylitis can have intestinal inflammatory lesions, thus the use of colonoscopy for such patients should be defined. Objectives: To assess the gross intestinal colonoscopic changes and microscopic histopathologic findings of patients with ankylosing spondylitis; to correlate the colonoscopic and histopathologic findings; and to study the relationship of the histopathologic findings with extra-articular manifestations of the disease, HLA-B27, BASFI and BASDAI. Methods: This is a cross-sectional study of 22 patients with ankylosing spondylitis. The patients underwent clinical assessment, BASDAI and BASFI application, blood collection for HLA-B27 measurement, and colonoscopy with biopsy of four intestinal segments (terminal ileum, right and sigmoid colons, and rectum). Results: Abnormal colonoscopic results were obtained in 13 (59.1%) patients, the major abnormality being intestinal polyps. The groups of normal and abnormal colonoscopic results (n = 9 and n = 13, respectively) were homogeneous regarding age, BASFI, BASDAI, and categorical variables, and the P-value showed no significant difference between groups. The histopathological findings revealed abnormal biopsies in 81%, 90.9%, 90.9% and 86.4% for terminal ileum, right colon, sigmoid colon, and rectum, respectively. The histopathologic results showed no statistically significant association with the extra-articular manifestations, BASFI, BASDAI and HLA-B27 positivity. Conclusions: The histological analysis of the four intestinal segments evidenced inflammatory lesions in patients with normal and abnormal colonoscopic results, independently of bowel symptomatology and therapy used in the treatment of the basal disease. .

Ecological study of gastric cancer in Brazil: geographic and time trend analysis.

AIM: To investigate the geographic distributions and time trends of gastric cancer (GC) incidence and mortality in Brazil. METHODS: An ecological study of the DATASUS registry was conducted by identifying hospitalizations for GC between January 2005 and December 2010. The data included information on the gender, age, and town of residence at the time of hospital admission and death. RESULTS: The GC rates, adjusted according to available hospital beds, decreased from 13.8 per 100000 in 2005 to 12.7 per 100000 in 2010. The GC rates decreased more among the younger age groups, in which the male-to-female difference also decreased in comparison to the older age groups. Although the lethality rates tended to increase with age, young patients were proportionally more affected. The spatial GC distribution showed that the rates were higher in the south and southeast. However, while the rates decreased in the central-west and south, they increased in the northern regions. A geographic analysis showed higher rates of GC in more urbanized areas, with a coast-to-inland gradient. Geographically, GC lethality overlapped greatly with the hospital admission rates. CONCLUSION: The results of this study support the hypothesis of a critical role for environmental factors in GC pathogenesis. The declining rates in young patients, particularly males, suggest a relatively recent decrease in the exposure to risk factors associated with GC. The spatial distribution of GC indicates an ongoing dynamic change within the Brazilian environment.

Multidrug resistance 1 gene polymorphisms may determine Crohn's disease behavior in patients from Rio de Janeiro.

OBJECTIVES: Conflicting data from studies on the potential role of multidrug resistance 1 gene polymorphisms in inflammatory bowel disease may result from the analysis of genetically and geographically distinct populations. Here, we investigated whether multidrug resistance 1 gene polymorphisms are associated with inflammatory bowel diseases in patients from Rio de Janeiro. METHODS: We analyzed 123 Crohn's disease patients and 83 ulcerative colitis patients to determine the presence of the multidrug resistance 1 gene polymorphisms C1236T, G2677T and C3435T. In particular, the genotype frequencies of Crohn's disease and ulcerative colitis patients were analyzed. Genotype-phenotype associations with major clinical characteristics were established, and estimated risks were calculated for the mutations. RESULTS: No significant difference was observed in the genotype frequencies of the multidrug resistance 1 G2677T/A and C3435T polymorphisms between Crohn's disease and ulcerative colitis patients. In contrast, the C1236T polymorphism was significantly more common in Crohn's disease than in ulcerative colitis (p = 0.047). A significant association was also found between the multidrug resistance 1 C3435T polymorphism and the stricturing form of Crohn's disease (OR: 4.13; p = 0.009), whereas no association was found with penetrating behavior (OR: 0.33; p = 0.094). In Crohn's disease, a positive association was also found between the C3435T polymorphism and corticosteroid resistance/refractoriness (OR: 4.14; p = 0.010). However, no significant association was found between multidrug resistance 1 gene polymorphisms and UC subphenotypic categories. CONCLUSION: The multidrug resistance 1 gene polymorphism C3435T is associated with the stricturing phenotype and an inappropriate response to therapy in Crohn's disease. This association with Crohn's disease may support additional pathogenic roles for the multidrug resistance 1 gene in regulating gut-microbiota interactions and in mediating fibrosis. Understanding the effects of several drugs associated with multidrug resistance 1 gene variants may aid in the selection of customized therapeutic regimens.

Thiopurine-methyltransferase variants in inflammatory bowel disease: prevalence and toxicity in Brazilian patients.

AIM: To analyze the prevalence of thiopurine-methyltransferase (TPMT) genotypes and their association with drug toxicity in inflammatory bowel disease (IBD) patients from southeastern Brazil. METHODS: A total of 219 consecutive patients with IBD, of which 146 had Crohn's disease and 73 had ulcerative colitis, regularly seen at the outpatient unit of the Division of Gastroenterology at the University Hospital Pedro Ernesto of the State University of Rio de Janeiro, a tertiary referral center, were enrolled in this study from February 2009 to January 2011. We analyzed the presence of major TPMT genetic variants (TPMT 2, 3A, 3C) in IBD patients by means of a specific allele and RFLP-PCR. Genomic DNA was isolated from peripheral blood leukocytes by proteinase-K/Sodium Dodecyl Sulfate digestion and phenol-chloroform extraction. TPMT 2 (C238G), TPMT 3A (G460A/A719G), and TPMT 3C (A719G) genotypes were detected by real-time polymerase chain reaction followed by direct sequencing with specific primers. Clinical data were systematically recorded, and correlated with the genotype results. RESULTS: The distribution of the selected TPMT gene polymorphism TPMT 2 (C238G), TPMT 3A (G460A/A719G), and TPMT 3C (A719G) genotypes was 3.6%, 5.4%, and 7.7% of the patients, respectively. Among the side effects recorded from patients taking azathioprine, 14 patients presented with pancreatitis and/or an elevation of pancreatic enzymes, while 6 patients had liver toxicity, and 2 patients exhibited myelosuppression/neutropenia. TPMT polymorphisms were detected in 37/219 patients (8 heterozygous for 2, 11 heterozygous for 3A, and 18 heterozygous for 3C). No homozygotic polymorphisms were found. Despite the prevalence of the TPMT 3C genotype, no differences among the genotype frequencies were significant. Although no association was detected regarding myelotoxicity or hepatotoxicity, a trend towards the elevation of pancreatic enzymes was observed for TPMT 2 and TPMT 3C genotypes. CONCLUSION: The prevalence of TPMT genotypes was high among Brazilian patients. Variants genes 2 and 3C may be associated with azathioprine pancreatic toxicity in a IBD southeastern Brazilian population.

Overexpression of ATP-activated P2X7 receptors in the intestinal mucosa is implicated in the pathogenesis of Crohn's disease.

BACKGROUND: Extracellular nucleotides released in conditions of cell stress alert the immune system from tissue injury or inflammation. We hypothesized that the P2X7 receptor (P2X7-R) could regulate key elements in inflammatory bowel disease pathogenesis. METHODS: Colonoscopy samples obtained from patients with Crohn's disease (CD), ulcerative colitis, and controls were used to analyze P2X7-R expression by RT and real-time PCR, immunohistochemistry, and confocal microscopy. Inflammatory response was determined by the levels of cytokines by enzyme-linked immunosorbent assay in cultures of intestinal explants. Apoptosis was determined by the TUNEL assay. P2X7-R C57BL/6 mice were treated with trinitrobenzene sulfonic acid or dextran sulfate sodium (DSS) for inducing colitis. RESULTS: P2X7-R was expressed in higher levels in inflamed CD epithelium and lamina propria, where it colocalizes more with dendritic cells and macrophages. Basal levels of P2X7-R mRNA were higher in CD inflamed mucosa compared with noninflamed CD and controls and were upregulated after interferon-γ in controls. Apoptotic rates were higher in CD epithelium and lamina propria compared with ulcerative colitis and controls. Levels of tumor necrosis factor-α, interleukin (IL)-1ß, and IL-17 were higher, whereas IL-10 was lower in CD compared with controls. Levels of tumor necrosis factor-α-α and interleukin-1ß increased after adenosine-triphosphate and decreased after KN62 treatment in CD. P2X7-R animals did not develop trinitrobenzene sulfonic acid or DSS colitis. CONCLUSIONS: The upregulation of P2X7-R in CD inflamed mucosa is consistent with the involvement of purinoceptors in inflammation and apoptosis. These observations may implicate purinergic signaling in the pathogenesis of intestinal inflammation, and the P2X7-R may represent a novel therapeutic target in CD.

[Colonoscopic evaluation in patients with ankylosing spondylitis]. / Avaliação colonoscópica em pacientes com espondilite anquilosante.

INTRODUCTION: Patients with ankylosing spondylitis can have intestinal inflammatory lesions, thus the use of colonoscopy for such patients should be defined. OBJECTIVES: To assess the gross intestinal colonoscopic changes and microscopic histopathologic findings of patients with ankylosing spondylitis; to correlate the colonoscopic and histopathologic findings; and to study the relationship of the histopathologic findings with extra-articular manifestations of the disease, HLA-B27, BASFI and BASDAI. METHODS: This is a cross-sectional study of 22 patients with ankylosing spondylitis. The patients underwent clinical assessment, BASDAI and BASFI application, blood collection for HLA-B27 measurement, and colonoscopy with biopsy of four intestinal segments (terminal ileum, right and sigmoid colons, and rectum). RESULTS: Abnormal colonoscopic results were obtained in 13 (59.1%) patients, the major abnormality being intestinal polyps. The groups of normal and abnormal colonoscopic results (n=9 and n=13, respectively) were homogeneous regarding age, BASFI, BASDAI, and categorical variables, and the P-value showed no significant difference between groups. The histopathological findings revealed abnormal biopsies in 81%, 90.9%, 90.9% and 86.4% for terminal ileum, right colon, sigmoid colon, and rectum, respectively. The histopathologic results showed no statistically significant association with the extra-articular manifestations, BASFI, BASDAI and HLA-B27 positivity. CONCLUSIONS: The histological analysis of the four intestinal segments evidenced inflammatory lesions in patients with normal and abnormal colonoscopic results, independently of bowel symptomatology and therapy used in the treatment of the basal disease.

Multidrug resistance 1 gene polymorphisms may determine Crohn's disease behavior in patients from Rio de Janeiro

OBJECTIVES: Conflicting data from studies on the potential role of multidrug resistance 1 gene polymorphisms in inflammatory bowel disease may result from the analysis of genetically and geographically distinct populations. Here, we investigated whether multidrug resistance 1 gene polymorphisms are associated with inflammatory bowel diseases in patients from Rio de Janeiro. METHODS: We analyzed 123 Crohn's disease patients and 83 ulcerative colitis patients to determine the presence of the multidrug resistance 1 gene polymorphisms C1236T, G2677T and C3435T. In particular, the genotype frequencies of Crohn's disease and ulcerative colitis patients were analyzed. Genotype-phenotype associations with major clinical characteristics were established, and estimated risks were calculated for the mutations. RESULTS: No significant difference was observed in the genotype frequencies of the multidrug resistance 1 G2677T/A and C3435T polymorphisms between Crohn's disease and ulcerative colitis patients. In contrast, the C1236T polymorphism was significantly more common in Crohn's disease than in ulcerative colitis (p = 0.047). A significant association was also found between the multidrug resistance 1 C3435T polymorphism and the stricturing form of Crohn's disease (OR: 4.13; p = 0.009), whereas no association was found with penetrating behavior (OR: 0.33; p = 0.094). In Crohn's disease, a positive association was also found between the C3435T polymorphism and corticosteroid resistance/refractoriness (OR: 4.14; p = 0.010). However, no significant association was found between multidrug resistance 1 gene polymorphisms and UC subphenotypic categories. CONCLUSION: The multidrug resistance 1 gene polymorphism C3435T is associated with the stricturing phenotype and an inappropriate response to therapy in Crohn's disease. This association with Crohn's disease may support additional pathogenic roles ...

Hedgehog pathway signaling regulates human colon carcinoma HT-29 epithelial cell line apoptosis and cytokine secretion.

The Hedgehog (Hh) pathway is involved in embryogenesis and physiologic processes including cell survival and proliferation. We used the HT-29 and other human colon carcinoma cell lines to investigate Hh signaling and biological functions in colonic epithelial cells. HT-29 cells were cultured under different conditions and exposed to various stimuli. The expression of Hh pathway components and related genes and proteins were assessed by real-time PCR and immunofluorescence. Viability, apoptosis and cell proliferation were measured by the MTT assay, Annexin-V/7-AAD staining and BrdU uptake, respectively. Chemokines production was measured by ELISA in culture supernatants. Indian and Sonic Hh mRNA levels and the downstream transcription factors Gli-1 and Gli-2 increased following treatment with Hh agonists and butyrate, but decreased upon exposure to cyclopamine or GANT61. BMP4 and BMP7 expression increased after stimulation with Hh agonists. Gli-1 protein expression increased after Hh agonists and decreased following cyclopamine. Exposure to Hh agonists promoted ß-catenin reduction and subcellular redistribution. Levels of IL-8 and MCP-1 decreased upon exposure to Hh agonists compared to Hh antagonists, LPS, IFN-γ or EGF. Monocyte chemotaxis decreased upon exposure to supernatants of HT-29 cells treated with Shh compared to Hh antagonists, LPS and IFN-γ. Cellular incorporation of BrdU and cell viability decreased following Hh blockade. Hh agonists abrogated the anti-CD95 induced apoptosis. Hh pathway is a key controller of colon cancer cells, as demonstrated by its effect in dampening inflammatory signals and antagonizing apoptosis. The differential expression of Hh components may underlie abnormalities in the local immune response and in epithelial barrier integrity, with potential homeostatic implications for the development of colonic inflammation and malignancies.

Prognostic significance of p53 protein expression in early gastric cancer.

Mutations of the p53 tumor suppressor gene have been associated with abnormalities in cell cycle regulation, DNA repair and synthesis, apoptosis, and it has been implicated in the prognosis of advanced gastric cancer. The aim of this study was to evaluate the occurrence of p53 gene mutation and its possible prognostic implications in early gastric cancer. In a retrospective study, we studied 80 patients with early gastric cancer treated surgically between 1982 and 2001. Mutation of p53 gene was investigated in surgical gastric specimens by immunohistochemistry, and results were analyzed in relation to gender, age, macroscopic appearance, size and location of tumor, presence of lymph nodes, Lauren's histological type, degree of differentiation, and the 5-year survival. The expression of p53 was more frequent among the intestinal type (p = 0.003), the differentiated (p = 0.007), and the macroscopically elevated tumors (p = 0.038). Nevertheless, the isolated expression of p53 was not associated with the 5-year survival, or with the frequency of lymph node involvement. The degree of differentiation was detected as an independent factor related to the outcome of patients (0.044). Significantly shorter survival time was found in p53-negative compared with p53-positive patients, when considering the degree of differentiation of tumors, as assessed by Cox regression analysis (0.049). The association of p53 with the intestinal type, the degree of differentiation and morphological characteristics, may reflect the involvement of chronic inflammatory process underlying early gastric cancer. In this population sample, the expression of p53 alone has no prognostic value for early gastric cancer. However, the significant difference in p53 expression between subgroups of degree of differentiation of tumors can influence post-operative outcome of patients and may be related to possible distinct etiopathogenic subtypes.

Prevalence of periodontitis and DMFT index in patients with Crohn's disease and ulcerative colitis.

AIM: To compare the prevalence of periodontal disease and the decayed, missing and filled teeth (DMFT) index in patients with Crohn's disease (CD) and ulcerative colitis (UC) with those without these diseases. MATERIAL AND METHODS: Ninety-nine CD (39.0 SD+/-12.9 years), 80 UC (43.3 SD+/-13.2) and 74 healthy controls (40.3 SD+/-12.9) were compared for DMFT index and presence of periodontitis. Probing pocket depth (PPD), clinical attachment loss (CAL), bleeding on probing (BOP), plaque and DMFT index were measured on all subjects. The presence of periodontitis was defined as having CAL > or =3 mm in at least four sites in different teeth. RESULTS: Significantly more patients with UC (90.0%; p<0.001) and CD (81.8%; p=0.03) had periodontitis than controls (67.6%). Among smokers, UC patients had significantly more periodontitis. CD had a greater mean DMFT score (18.7 versus 13.9; p=0.031) compared with controls and UC had greater median PPD (2.2 versus 1.7 mm; p<0.0001) than controls. Among non-smokers, CD (2.4 mm; p<0.0001) and UC showed deeper pockets (2.3 mm; p<0.0001) compared with controls (1.5 mm). UC had a greater mean DMFT score (15.3 versus 12.1; p=0.037) compared with controls. CONCLUSIONS: CD and UC patients had higher DMFT and prevalence of periodontitis than controls, but smoking was an effect modifier.

Prevalência de periodontite em pacientes com doença inflamatória intestinal / Prevalence of periodontits in patients with inflammatory bowel disease

O objetivo deste estudo foi avaliar a prevalência de periodontite em pacientes com doenças inflamatórias intestinal (DII), e compará-los a pacientes saudáveis sistemicamente. Foram examinados 99 pacientes com Doença de Crohn (DC) (39 DP ± 12,9 anos), 80 com Retocolite Ulcerativa (RCUI) (43,3 ± 13,2 anos) e 74 no grupo controle (C ) (40,3 ± 12,9 anos). A condição periodontal foi avaliada através da placa visível, do sangramento à sondagem, da profundidade de sondagem (OS) e do nível clínico de inserção (NCI). Indivíduos que apresentavam pelo menos quatro sítios com NCI ≥ 3 mm foram considerados como portadores de periodontite. Havia mais pacientes com periodontite nos grupos RCUI (92,6%, p=0,004) e DC (91,9%, p=0,019), comparado ao grupo C (79,7%). O grupo com DC apresentava significativamente menos placa (p=0,017), menos sangramento à sondagem (p-0,038) e menos sítios com os ≥ 4mm (p=0,02) quando comparado ao grupo controle. Nenhuma relação de significância ocorreu nestes índices entre os grupos RCUI e C. Conclui-se que tanto os pacientes com DV quanto com RCUI apresentaram maior prevalência de periodontite do que o grupo controle.