Ovarian Reserve Assessed by the Anti-Mullerian Hormone and Reproductive Health Parameters in Women With Crohn´s Disease, a Case-Control Study.

According to several studies, women with Crohn's disease (CD) had reduced fertility, which is mostly due to voluntary decisions and reduced ovarian reserve. In our study, we aimed to compare reproductive health parameters (RHP), previous pregnancy complications and outcomes, and ovarian reserve (OR) assessed by the anti-Mullerian hormone (AMH) in CD patients with healthy controls. In CD patients, we also compared OR according to disease phenotypes. Consecutive pre-menopausal women with CD from two IBD centers were included. The control group consisted of age and BMI-matched healthy controls. We used a questionnaire that included RHP, CD phenotype, and CD activity. Serum AMH was assessed by the Elecsys AMH plus essay. We enrolled 50 patients and 56 controls with a median age of 31 years. All CD patients were in clinical remission. We observed no difference in RHP or AMH (median 2.6 vs. 2.1 ug/l, p = 0.98), or the proportion of low OR (AMH<1,77, 38 vs. 41.1 %, p=0.84). The slope of age-related decrease did not differ between the groups. The subgroup of CD patients after surgery and those older than 30 years with CD for >5years had a steeper decrease in AMH (slope -0.12 vs. -0.29, p = 0.04 and -0.31 vs. -0.2, p = 0.029). In a multivariate analysis, age was the single independent predictor of low OR (OR=1.25). In women with Crohn's disease, once the disease activity is under control, the reproductive health and ovarian reserve do not substantially differ from healthy controls.

Ovarian Reserve Assessed by the Anti-Mullerian Hormone and Reproductive Health Parameters in Women With Crohn´s Disease, a Case-Control Study.

According to several studies, women with Crohn's disease (CD) had reduced fertility, which is mostly due to voluntary decisions and reduced ovarian reserve. In our study, we aimed to compare reproductive health parameters (RHP), previous pregnancy complications and outcomes, and ovarian reserve (OR) assessed by the anti-Mullerian hormone (AMH) in CD patients with healthy controls. In CD patients, we also compared OR according to disease phenotypes. Consecutive pre-menopausal women with CD from two IBD centers were included. The control group consisted of age and BMI-matched healthy controls. We used a questionnaire that included RHP, CD phenotype, and CD activity. Serum AMH was assessed by the Elecsys AMH plus essay. We enrolled 50 patients and 56 controls with a median age of 31 years. All CD patients were in clinical remission. We observed no difference in RHP or AMH (median 2.6 vs. 2.1 ug/l, p = 0.98), or the proportion of low OR (AMH<1,77, 38 vs. 41.1 %, p=0.84). The slope of age-related decrease did not differ between the groups. The subgroup of CD patients after surgery and those older than 30 years with CD for >5years had a steeper decrease in AMH (slope -0.12 vs. -0.29, p = 0.04 and -0.31 vs. -0.2, p = 0.029). In a multivariate analysis, age was the single independent predictor of low OR (OR=1.25). In women with Crohn's disease, once the disease activity is under control, the reproductive health and ovarian reserve do not substantially differ from healthy controls.

Novel unorthodox strategies to reduce the case fatality rate of COVID-19 in high risk groups.

Dear editors: We propose novel strategies to combat the COVID-19 outbreak, that are aimed at high-risk groups and might reduce the progression to severe forms of COVID and thus decrease the very high case fatality rate.Following the first reports of the outbreak of several cases of acute respiratory distress syndrome in the Chinese city of Wuhan at the end of December 2019, a novel beta coronavirus, named severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2 or 2019-nCoV) as the main causative agent was identified while the disease associated with was named by WHO as COVID-19 (1, 2). The outbreak has rapidly spread globally with more than 150.000 cases detected in over 100 countries as of March 13, 2020. The overall case-fatality rate (CFR) of COVID-19 in China was 2.3 %, but globally it seems to be higher in the range of 3‒5 % (3). At present, no specific antivirals or approved vaccines are available to combat COVID-19. Many patients however receive off-label antivirals such as lopinavir/ritonavir, ribavirin and/or chloroquine and /or interferons. Several new antivirals such as remdesivir are studied in ongoing clinical trials. Given the alarming global situation and rapidly evolving large scale pandemics, there is an urgent need for effective strategies to prevent the spread of the disease and decrease its high CFR. The gravity of the situation requires to consider even novel unorthodox strategies to control the outbreak and high lethality of COVID-19. (Tab. 2, Ref. 21). Keywords: COVID-19, coronavirus, distress syndrome, lopinavir/ritonavir, ribavirin, chloroquine, interferons.

Pathophysiology and risk factors for cholelithiasis in patients with Crohn's disease.

Cholelithiasis is more common in patients with Crohn's disease (CD) than in the healthy population. The aim here was to examine risk factors for cholelithiasis in a cohort of CD patients and to compare the prevalence of cholelithiasis in a cohort of CD patients with that in a control group. This was a single-center retrospective case-control study. The cohort comprised all consecutive CD patients who underwent abdominal ultrasound from January 2007 to January 2018. The control group comprised age- and gender-matched non-CD patients referred for upper gastrointestinal tract dyspepsia. The study included 238 CD patients and 238 controls. The prevalence of cholelithiasis in the CD and control groups was 12.6 % and 9.2 %, respectively (risk ratio (RR), 1.36; p=0.24). Univariate analysis revealed that cholelithiasis was associated with multiple risk factors. Multivariate analysis identified age (OR, 1.077; 95 % CI, 1.043-1.112; p<0.001) and receipt of parenteral nutrition (OR, 1.812; 95 % CI, 1.131-2.903; p=0.013) as independent risk factors for cholelithiasis in CD patients. The prevalence of cholelithiasis in CD patients was higher than that in the control group; however, the difference was not statistically significant. Age and receipt of parenteral nutrition were independent risk factors for cholelithiasis in CD patients.

Impact of smoking cigarette on the mRNA expression of cytokines in mucosa of inflammatory bowel disease.

It is well known that smoking is the risk factor in the development and clinical course of Crohn s disease (CD), but on the other hand, smoking is a protective factor against ulcerative colitis (UC). The pathways that are influenced by smoking in CD and UC are poorly understood. The aim of our study was to analyse the influence of smoking on the mRNA expression of cytokines in mucosa in patients with CD and UC. We performed a cross-sectional study. The cohort consisted of 86 IBD patients (48 CD patients and 38 UC patients) and took place at the IBD Centre at the University Hospital Bratislava-Ruzinov. We took the demographic and clinical data of each patient, including information about their smoking habits. We performed a colonoscopy on each patient and took biopsies from both inflamed and non-inflamed sigma (CD, UC) and terminal ileum (CD). mRNA was extracted from mucosal biopsy samples for each cytokine and was normalized to a housekeeping gene (GAPDH). Finally, we compared the mRNA expression of target cytokines in the mucosa of smokers and non-smokers in IBD patients. Smokers with Crohn s disease have a significantly higher mRNA expression of pro-inflammatory cytokine TNF ? (p=0.003) in inflamed mucosa in sigma compared with non-smokers. In smokers with ulcerative colitis, we observed significantly higher mRNA expression of anti-inflammatory cytokine IL 10 (p=0.022) in non-inflamed mucosa of sigma. Similarly, smokers with UC have a significantly decreased mRNA expression of cytokine TLR 2 (p=0.024) and CCR1 (p=0.049) in non-inflamed mucosa of sigma. Based on our results, smoking has a positive influence on cessation and the clinical course of UC due to the stimulation of anti-inflammatory cytokine IL 10 in mucosa. On the other hand, smokers with CD have a higher expression of pro-inflammatory cytokine TNF ?, which could be associated with a worsening of the disease and response to therapy.

Influence of vitamin D on the expression of mRNA of cytokines in the mucosa of inflammatory bowel disease patients.

The aim of this study was to analyze the influence of 25(OH)VD serum concentration on the expression of mRNA cytokines (IL-6, IL-8, IL-12, IL-17, IL-23, TNFα, CCR1, CCR2, CCR5, CCR9, CCL5, TLR2, TLR4, TLR5, CD207 ,CD206, FoxP3) in mucosa of IBD patients. The cohort consisted of 86 IBD patients (48 CD and 38 UC) followed at the IBD center of University Hospital Bratislava-Ruzinov. We performed colonoscopy in each patient and took biopsies from mucosa of sigma and terminal ileum. Serum concentration of 25(OH)VD was assessed at the time of colonoscopy. mRNA was extracted from mucosal biopsy samples for each cytokine. Then we analyzed the correlation between VD and the expression of mRNA of cytokines from biopsies samples.  In CD we observed a significant positive correlation of serum concentration 25(OH)VD and the expression mRNA level of IL-6. There was also trend towards significant positive correlation of the expression mRNA of TNFα, IL-10, IL-23, TLR 2 in inflamed mucosa of terminal ileum as well as the expression mRNA of CCR5 and CCR1 in non-inflamed mucosa from terminal ileum. We also found a trend towards positive correlation between 25(OH)VD and the expression mRNA of IL-23, TLR4, CD 207, CCR1, CCR5 and CD 206 in non-inflamed mucosa of sigma in UC.VD significantly correlated with the levels of expression of several inflammatory cytokines including TNFα in colonic mucosa of patients with IBD (Tab. 4, Fig. 3, Ref. 31).

Dosing interval and diagnosis predict infliximab levels in patients with inflammatory bowel disease on maintenance treatment.

OBJECTIVE: The aim of the study was to identify factors influencing infliximab (IFX) trough levels (TL) in patients with inflammatory bowel disease (IBD). METHODS: This was a multicentre cross-sectional study performed at 5 large IBD centres in Slovakia. The cohort consisted of IBD patients, treated either with original IFX or CT-P13 biosimilar, who were examined for the IFX TL and antidrug antibodies (ADA) in a central laboratory. RESULTS: The patient cohort consisted of 116 consecutive IBD patients, 68 with Crohn's disease (CD) and 48 with ulcerative colitis (UC). CD patients had significantly lower IFX TL compared to UC, 2.41 (0.998-5.56) mg/L vs. 4.49 (1.76-8.41) mg/L, p = 0.017. During maintenance treatment, significantly higher mean IFX TL were observed in patients with a 4 week dosing interval than in patients with a 6 or 8 (7.44±3.6 µg/mL vs. 4.19±4.2 vs. 3.30±3.1 µg/mL, p = 0.011 and p< 0.0001, respectively). There was no difference in median TL IFX between original IFX and biosimilar CT-P13 (3.25 (1.24-6.52) mg/L vs. 3.03 (1.30-7.10)). IFX TL correlated with ADA (p=0.005). Multiple regression analysis revealed two independent factors for IFX TL: dosing interval (p<0.0001) and diagnosis (p=0.02). CONCLUSION: In the present study we observed that IBD patients assigned to an intensified dosing interval during maintenance therapy have significantly higher IFX TL than patients receiving conventional 8 week interval. Patients with UC had significantly higher IFX TL.

Relapse rates of inflammatory bowel disease patients in deep and clinical remission after discontinuing anti-tumor necrosis factor alpha therapy.

BACKGROUND AND AIMS: Relapse rates after discontinuing anti-tumor necrosis factor-α (TNFα) therapy of inflammatory bowel disease (IBD) patients in deep remission are poorly understood. This prospective single-center open-label study evaluated the relapse rates of IBD patients after stopping anti-TNFα therapy. METHODS: All IBD patients who were in clinical remission and stopped anti-TNFα therapy in 2011-2013 and were followed up for at least 12 months were enrolled. The "Ultradeep" patients were in calprotectin-negative (<50 ng/g) deep remission for at least six months and ceased anti-TNFα therapy on physician recommendations. The "clinical" patients were in clinical but not deep remission and ceased anti-TNFα therapy for other reasons. Relapse rates were assessed and relapse risk factors identified. RESULTS: One year after stopping, 27 % and 27 % of the Ultradeep (n = 11) and Clinical (n = 11) patients relapsed, respectively. Two years after stopping, 57 % and 62 % relapsed, respectively (p = 0.89). All relapsed patients who underwent retreatment with anti-TNFα therapy re-entered remission. Male sex was a significant risk factor for relapse (p = 0.03). CONCLUSION: Our study showed that even highly selected IBD patients who lack clinical, endoscopic or laboratory signs of disease activity have a relatively high relapse rate in the follow-up period after ceasing anti-TNFα therapy (Tab. 2, Fig. 3, Ref. 24).

[Intramural esophageal dissection]. / Intramurálna disekcia pazeráka.

The intramural esophageal dissection is a rare condition with benign course, good prognosis and low mortality. It is difficult to differentiate it from an esophageal perforation at the presentation time. The conservative therapy should be considered as the treatment of choice in most cases. Complications such as esophageal perforation, massive bleeding or persistence of the symptoms are well accepted indications for surgical procedure. Radical treatment in the form of esophagectomy is extremely rare and it is usually indicated only for patients with complicated esophageal perforation. The authors offer two cases with female patients suffering from the intramural esophageal dissection who were indicated for esophagectomy without any proven esophageal perforation.

The prevalence and risk factors for osteoporosis in patients with inflammatory bowel disease.

AIM: Osteoporosis is a known chronic complication of inflammatory bowel diseases (IBD). The aim of our study was to describe the prevalence of reduced bone mineral density (BMD) in IBD patients and to identify crucial risk factors for osteoporosis. METHODS: The cohort consisted of 76 IBD patients, 40 with Crohn's disease (CD) and 36 with ulcerative colitis (UC). Clinical characteristics of every patient were recorded, i.e. age, sex, duration of the disease, clinical behavior, location of disease according to Montreal classification, surgeries, steroid medication, sIBDQ, and smoking habits. We examined the serum 25-hydroxyl vitamin D3 (25-OHD3) in each patient. The BMD was determined by dual-energy X-ray absorptiometry (DXA) at the lumbar spine and femoral neck. RESULTS: Osteoporosis was documented in 10 IBD patients (13.2 %), while osteopenia in 35 of them (46.1 %). Patients with CD have significantly lower femoral Z score than patients with UC. Femoral Z score was strongly associated with disease duration, and in CD patients suffering from stricturing form, with ileic or ileocolic location and history of proctocolectomy or total colectomy. Patients with osteoporosis had a significantly lower level of 25-OHD3 than patients with normal BMD. CONCLUSION: Patients with long disease duration and those suffering from stricturing form of CD with ileic/ileocolic location and history of proctocolectomy/total colectomy are at higher risk of developing osteoporosis than other IBD patients. The high proportion of osteopenia/osteoporosis in our study underlines the importance of BMD measurement in all IBD patients as a base for initiating the appropriate treatment (Tab. 1, Fig. 3, Ref. 63).

Prevalence of mutations in thiopurine S-methyltransferase gene among Slovak IBD patients.

BACKGROUND: Thiopurine S-methyltransferase (TPMT) plays an important role in the metabolism of thiopurines. It has been suggested that TPMT genetic polymorphisms lead to dose-related hematopoietic toxicity. Since there are major ethnic differences in the prevalence of particular TPMT variants, it is important for each country to study their own prevalence in order to estimate the role of TPMT variants-related thiopurines toxicity in population suffering from particular inflammatory bowel disease (IBD). AIMS: The aim of this study was to determine the frequency of the four most common allelic variants of TPMT gene in the population of Slovak IBD patients. METHODS: TPMT genetic polymorphisms (TPMT*2, TPMT*3A, TPMT*3B, TPMT*3C) were amplified using PCR and consequently genotyped with genetic analyzer. The allele frequencies of particular allelic variants were calculated and compared with other Caucasian populations reported so far. RESULTS: Three hundred and thirty IBD patients were included; 196/132/2 cases of Crohn´s disease/ulcerative colitis/unclassified colitis; 180 (55 %) males. Ninety-three percent of patients were homozygous for wild-type TPMT variant. Heterozygous genotype of any of the studied polymorphisms was present in 6 % of patients while only one patient was homozygous for TPMT*3A allele (0.3 %). The most prevalent mutant allele was that of TPMT*3A (3.2 %). The distribution of most common allelic variants of TPMT gene among Slovak IBD patients was in accordance with previously reported prevalence in Caucasian populations. CONCLUSION: This study shows the prevalence of TPMT genetic polymorphisms in population of Slovak IBD patients. As in other Caucasian populations, the most common mutant allelic variant is that of TPMT*3A while the prevalence of homozygosity is relatively low (Tab. 3, Ref. 22).

The impact of thiopurine-S-methyltransferase genotype on the adverse drug reactions to azathioprine in patients with inflammatory bowel diseases.

BACKGROUND AND AIMS: The thiopurine drugs, azathioprine (AZA) and 6-mercaptopurine, are established in the treatment of inflammatory bowel diseases (IBD). Polymorphisms in thiopurine S-methyltransferase (TPMT) gene have been associated with adverse drug reactions (ADRs) to AZA. METHODS: The aim of this study was to evaluate TPMT polymorphisms and AZA-related toxicity in a Slovak cohort of 220 IBD patients treated with AZA. In every patient, the dose and duration of AZA therapy, concomitant 5-aminosalicylate (5-ASA) medication, frequency, type, time to onset, dose of ADR and concomitant 5-ASA at the onset of ADR were recorded. Each patient was also genotyped for the presence of variant TPMT alleles (*2,*3A,*3B,*3C). Frequency, type and circumstances of ADRs were compared according to TPMT status. RESULTS: Of the 220 patients, 205 (93.2 %) were wild-type (TPMT*1/*1), one (0.5%) carried a TPMT*1/*3C allele, 13 (5.9 %) carried TPMT *1/*3A allele and one was homozygous for TMPT *3A allele. No TPMT *2 mutation was found. The incidence of adverse drug reactions was 62/205 (30.2 %) in the wild-type group as compared to 13/15 (86.7 %) in the TPMT mutation group, p=2.10-5. Leukopenia (WBC< 3.0*10^9/L) occurred in 21/205 (10.2 %) patients with wild type TPMT versus 11/15 (73.3 %) patients with TPMT mutations, p=0.000001. There was no significant difference between TMPT groups in gastrointestinal or other ADRs. No impact of 5-ASA on the incidence and severity of AZA adverse drug reactions was observed. CONCLUSION: The incidence of leukopenia in TPMT mutant patients was significantly higher and more severe as compared to TPMT wild type patients. We observed no impact of concomitant 5-ASA therapy on AZA induced toxicity (Tab. 4, Fig. 2, Ref. 37).

Prevalence of mutations in thiopurine S-methyltransferase gene among Slovak IBD patients.

BACKGROUND: Thiopurine S-methyltransferase (TPMT) plays an important role in the metabolism of thiopurines. It has been suggested that TPMT genetic polymorphisms lead to dose-related hematopoetic toxicity. Since there are major ethnic differences in the prevalence of particular TPMT variants, it is important for each country to study their own prevalence in order to estimate the role of TPMT variants-related thiopurines toxicity in the particular inflammatory bowel disease (IBD) population. AIMS: The aim of this study was to determine the frequency of the four most common allelic variants of TPMT gene in the population of Slovak IBD patients. METHODS: TPMT genetic polymorphisms (TPMT*2, TPMT*3A, TPMT*3B, TPMT*3C) were amplified using PCR and consequently genotyped on genetic analyzer. The allele frequencies of particular allelic variants were calculated and compared with other Caucasian populations reported so far. RESULTS: Three hundred and thirty IBD patients were included; 196/132/2 Crohn´s disease/ulcerative colitis/unclassified colitis, 180 (55 %) males. Ninety-three percent of patients were homozygous for wild type TPMT variant. Heterozygous genotype of any of the studied polymorphisms was present in 6 % of patients, only one patient was homozygous for TPMT*3A allele (0.3 %). The most prevalent mutant allele was TPMT*3A (3.2 %). The distribution of the most common allelic variants of TPMT gene among Slovak IBD patients were in accordance with previously reported prevalence in Caucasian populations. CONCLUSION: This study shows the prevalence of TPMT genetic polymorphisms in the Slovak IBD patient`s population. As in other Caucasian populations, the most common mutant allelic variant is TPMT*3A, and the prevalence of homozygosity is relatively low (Tab. 3, Ref. 16).

Detection of point mutations in KRAS oncogene by real-time PCR-based genotyping assay in GIT diseases.

OBJECTIVES: The determination of gene mutations is important for the diagnosis and prognosis of various gastrointestinal cancers. The aim of our study was to develop a new procedure for the analysis of KRAS gene mutation by application of the real-time PCR method. BACKGROUND: The detection process requires discriminate trace amount of mutant allele in a large excess of wild-type DNA in various samples. METHODS: The real-time PCR based technique using hybridization probes for five most frequently KRAS codon 12 mutations and WT specific peptide nucleic acid (PNA) was performed. Our multiplex detection system was tested in various DNA samples (tissue, bile, pancreatic juice) of patients with different diagnoses of gastrointestinal tract disease obtained by endoscopy and ERCP. RESULTS: We designed and optimized the real-time PCR conditions and tested various amount of PNA in PCR reaction to suppress amplification of the wild-type DNA. We determined the interassay variability of the melting temperatures and the results of mutation testing were confirmed by DNA sequencing with the 100 % accuracy. Incidence of searched mutations was 67.5 % in cohort of 40 patients; for KRASG12D it was in 44.4 %, KRASG12V in 22.2 %, KRASG12S in 14.8 %, KRASG12A in 14.8 % and KRASG12C in 3.8 %. The sensitivity of the assays is 1x10-5. CONCLUSIONS: Advantages of this technique are rapidity, accuracy and it is generally easy to perform. This method can be adapted for synchronic detection of multiple mutations and after readjustment by other type mutation of KRAS gene may serve as useful clinical tool for analyzing point mutations in various clinical samples (Tab. 3, Fig. 3, Ref. 42).

[Prevalence of liver disease markers among patients with metabolic risk factors]. / Prevalencia markerov polkodenia pecene u pacientov s metabolickými rizikovými faktormi.

INTRODUCTION: Non-alcoholic fatty liver disease is one of the most common liver diseases. It's prevalence among patients with metabolic risk factors (obesity, type 2 diabetes, hypertension, lipid disorders) without previously recognized liver disease is not completely known. Aims of our study were to determine the prevalence of liver lesions (elevated alanin aminotransferase (ALT), gamma glutamyl transpeptidase (GGT) above normal range and ultrasound signs of liver steatosis) among the study group of patients with at least one metabolic risk factor, to compare it with the control group with no risk factor, to investigate it's association with the number of metabolic risk factors and to identify it's closest independent predictors. METHODS: Patients with other known liver diseases were excluded. Among 482 patients 429 were in the study group and 53 in the control group. RESULTS: In the study group the prevalence ofALT, GGT elevation and signs ofsteatosis was 12.1, 29.9, 38.3%, comparing to 5.7, 11.9 and 5.7% in the control group respectively. The differences were statistically significant. With the increasing number of risk factors we found growing prevalence of GGT elevation and signs of steatosis, but ALT elevation was equally prevalent. In multiple logistic regression the only independent predictor of ALT elevation was obesity, predictors of GGT elevation were type 2. diabetes and signs of steatosis, signs of steatosis were independently associated with overweight, obesity, type 2 diabetes and hypertriglyceridemia. CONCLUSIONS: Markers of liver disease do have a clinical and prognostic impact on the liver and cardiometabolic risk and therefore we suggest they should be actively screened in this group of patients.

[Impact of basal prolactin levels on the prevalence of complications and the prognosis of patients with liver cirrhosis]. / Vplyv bazálnej koncentrácie prolaktínu na výskyt komplikácií a na prognózu pacientov s cirhózou pecene.

INTRODUCTION: Prolactin concentration was proved to be associated with complications of cirrhosis. METHODS: This relationship was investigated on the group of 90 patients predominantly males with alcoholic cirrhosis. Basic clinical and laboratory data were collected at entry as well as complications of cirrhosis already present. Patients were followed for the mean of 434 days and events such as variceal bleeding, hepatorenal syndrome and death were recorded. RESULTS: We found that 16.7% of patients had elevated serum prolactin levels, and had significantly higher Child-Pugh and MELD scores as well as higher ascites and encephalopathy stage. By comparing prolactin concentration quartiles we found complications of cirrhosis such as ascites, higher INR, jaundice and higher Child-Pugh and MELD scores more often with increasing prolactin concentrations. The most prominent was the relationship to hepatic encephalopathy (0 vs 31% between the 1st and 4th quartile, p < 0.05) to which the prolactin levels above 10.5 microg/l had a sensitivity of 92.9% and negative predictive value of 97%. Kaplan-Meier survival curve showed that patients in the 1st comparing to the 4th quartile had significantly higher survival rates (85.2 vs 50%, p = 0.046, hazard ratio = 0.2881). Prolactin levels > 11.91 microg/l had 80.8% sensitivity and 87.8% negative predictive value to predict death during the follow up period. CONCLUSION: Basal prolactin concentration could therefore be used as an alternative marker of hepatic encephalopathy and death in a selected subset of patients with cirrhosis.

Assay design and optimization of mutant-enriched PCR based method for detection of K-ras gene mutations in pancreatic carcinoma.

The aim of our work was to develop a fast, reliable and sensitive PCR method to detect K-ras mutations in various clinical samples. There is a need for an unimpeachable method for early diagnosis and/or screening of pancreatic cancer (PC). We optimized and subsequently analyzed four methods based on mutant-enriched PCR for the sensitivity, cost and time expense. Using the selected optimal method we examined codon 12 K- ras mutations in a study population of 59 patients with upper GIT malignancies. Reliability of the genotyping was confirmed by sequencing. By using the best of our modified mutant-enriched PCR methods we achieved sensitivity of 1:1 x 10(5). Further studies are necessary to determine the optimal biological material sampling in PC.

Polymorphisms in apoptosis genes predict response to infliximab therapy in luminal and fistulizing Crohn's disease.

BACKGROUND: Infliximab treatment is effective in 70-80% of patients with refractory luminal and fistulizing Crohn's disease. The effect of infliximab is ascribed to induction of apoptosis. AIM: To study whether polymorphisms in apoptosis genes predict the response to infliximab and whether they interact with clinical predictors. METHODS: Cohort of 287 consecutive patients treated with infliximab for refractory luminal (n = 204) or fistulizing (n = 83) Crohn's disease was genotyped for 21 polymorphisms in apoptosis genes. Short-term clinical response was assessed at week 4 (luminal Crohn's disease) or 10 (fistulizing Crohn's disease) after the first infliximab infusion. RESULTS: The response rate was 69% in luminal and 80% in fistulizing Crohn's disease. In luminal Crohn's disease, two genetic predictors were identified: (i) patients with the Fas ligand -843 CC/CT genotype (n = 135) responded in 75%, with the TT genotype (n = 21) in 38% only (P = 0.002; OR = 0.11; 95% CI: 0.08-0.56). (ii) Patients with the caspase-9 93 TT (n = 9) genotype all responded, in contrast with 67% (n = 147) with the CC and CT genotype (P = 0.04; OR = 1.50; 95% CI: 1.34-1.68). Concomitant azathioprine/mercaptopurine therapy overcame the effect of unfavourable genotypes. In the fistulizing Crohn's disease cohort, the same Fas ligand -843 CC/CT genotype was the only predictor of response (P = 0.002; OR = 1.66; 95% CI: 1.21-2.29), interacting with caspase-9 93 polymorphism but not with azathioprine/mercaptopurine. CONCLUSION: We observed that polymorphisms in FasL/Fas system and caspase-9 influence the response to infliximab in luminal and fistulizing Crohn's disease. The strongest association was seen between the Fas ligand -843 TT genotype and non-response. Concomitant mercaptopurine/azathioprine therapy, however, was able to overcome the effect of unfavourable genotypes in luminal disease.

The impact of major depressive disorder on the short- and long-term outcome of Crohn's disease treatment with infliximab.

BACKGROUND: Major depressive disorder is the most common psychiatric diagnosis in Crohn's disease. In other chronic diseases, evidence suggests that depression influences the course of the disease. Strong evidence of such a mediating role of major depressive disorder in Crohn's disease has never been found. AIM: To assess the relationship between major depressive disorder and outcome of treatment of luminal Crohn's disease with infliximab. METHODS: In this prospective study, 100 consecutive unselected patients underwent assessment of psychosocial, demographical disease-related biological and clinical parameters at baseline and at 4 weeks after infliximab. Major depressive disorder was diagnosed using the Patient Health Questionnaire. Subsequently, the patients were followed up clinically until the next flare or during 9 months. RESULTS: The Crohn's disease responded in 75% of the patients, and remission was achieved in 60%. The presence of major depressive disorder at baseline predicted a lower remission rate (OR = 0.166, 95% CI = 0.049-0.567, P = 0.004). At follow-up, 88% of the patients needed retreatment. At univariate regression analysis, major depressive disorder significantly decreased time to retreatment (P = 0.001). Multivariate Cox regression confirmed major depressive disorder as an independent determinant of active disease both at baseline and at re-evaluation (hazard ratio = 2.271, 95% CI: 1.36-3.79, P = 0.002). CONCLUSION: Major depressive disorder is a risk factor for failure to achieve remission with infliximab and for earlier retreatment in patients with active luminal Crohn's disease. Assessment and management of major depressive disorder should be part of the clinical approach to patients with Crohn's disease.

Positive family history promotes participation in colorectal cancer screening.

BACKGROUND: Participation rates in colorectal cancer (CRC) screening are rather low. We evaluated the interest of first degree relatives (FDR) of CRC patients to participate in a colonoscopy screening and compared the findings to controls with a negative family history. METHODOLOGY: There were 235 CRC patients diagnosed in our centre in 1984-2001. These were mailed an invitation letter for a preventive examination for their FDR older than 40 years and a questionnaire about occurrence of malignancies in their family. Colonoscopy was performed in 52 FDR and sex/age matched controls. RESULTS: The questionnaire was delivered to 196 patients. Thirty four (17.3%) patients responded. Positive family history for CRC was reported in 12/34 (35.3%) patients, compared to expected 3.4 patients (p = 0.04; OR 4.2; 95% CI = 1.05-17.89). Fifty two of 94 (55.3%) FDR participated in a screening and CRC was diagnosed in 2 and CRA in 18 patients compared to 1 CRC and 9 CRA in control group (p = 0.04; Kaplan-Meier p = 0.04). CONCLUSIONS: Positive family history seems to be a motivation factor for a participation in a CRC screening program. Consistent with previous studies the prevalence of CRA and CRC was significantly higher in the group of FDR compared to controls (Tab. 3, Fig. 1, Ref. 20).